Table of Content

24 June 2007, Volume 28 Issue 3

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Effects of Amyloid Precursor Protein 17 Peptide on the Hippocampal Neuron-insulin Signaling Transduction Pathway of Diabetic Kkay Mice

Zhuang Xiaoming;Mu Jun;Li Ran;Chen Min;Wang Rong;Ji Zhijuan;Zhao Zhiwei;Sheng Shuli

2007, 28(3):  273-277. 

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Objective Amyloid precursor protein(APP) is one in which neurotrophic and neurotoxic functions occur on the same chain.The APP319-335 segment(APP17 peptide) can enhance neuritic growth.Solube APP17 peptide exerts its effect through binding to specific binding sites on the cell surface.The present work is to investigate the effect of APP17 peptide on the expressions of insulin signaling transduction protein of T2 diabetic KKAy mice hippocampal neurons.Methods KK mice were divided into three groups: a normal control group(C,n=10),a diabetic group(DM,n=10) and an APP17mer peptide-protected diabetic group(DM+APP17P,n=10).Blood glucose in a tail-vein sample was determined by a glucose analyzer.A value>15 mmol/L was accepted as a diabetic model.APP17mer peptide was synthesized by solid phase method and purified by HPLC.Mice in the APP17+DM group were injected peptide subcutaneously at the dose of 8 μg/kg three times a week for 12 weeks.After twelve weeks,all the mice were killed and peripheral blood were obtained to assess the levels of HbA1c,plasma insulin and blood glucose.Homogenates were prepared from mice hippocampal tissue.Immunoprecipitation,Western-blot and immunohistochemistry were used to analyze the proteins related to insulin signaling transduction pathway.Tunnel assay was used to detect the apoptotic cell in hippocampus of KK mice.Immunohistochemical assay was performed with insulin receptor(InsR),insulin receptor substrate-1(IRS-1) and Bcl-2.Results The results of InsR and IRS-1 immunohistochemical staining showed that the hippocampus of C group and DM+APP17P contained darkly stained InsR and IRS-1positive cells.In contrast positive reacting cells in the hippocampus of DM group mice were pale,and were visibly fewer than in those Cgroup and APP17P+DM group mice. The number of positively stained hippocampal cells for Bcl-2 was similar in three groups.The expressions of CREB,phosphorylated CREB(p-CREB) and Bcl-2 showed no significant difference among the three groups.No positive Tunel cell was found in all groups.Conclusion Abnormal signal transduction pathway is present in hippocampal neuron of kk mice.The abnormality may not lead to apoptosis due to auto-regulation of neurons to a certain extent.This abnormal insulin signaling transduction pathway can be improved by APP17 peptide.

Expression of ACE-related Carboxypeptidase(ACE2) in Human and Rodent Pancreata

Ji Xiujuan;Xu Jian;Zhang Xuelian;Guo Limin;Yang Jinkui

2007, 28(3):  278-282. 

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Objective ACE-related carboxypeptidase(Angiotensin-converting Enzyme 2,ACE2),an enzymatic homologue of ACE,has been recently demonstrated as a functional receptor for severe acute respiratory syndrome coronavirus(SARS-CoV).Our previous study found that blood glucose levels in patients with SARS were significantly higher than control,which suggested that SARS-CoV might attack and cause damage to the pancreas.ACE2 enzyme plays a biological role in the degradation of angiotensin II,which provides a rationale for the further exploration of its role in many pathophysiological processes.In most tissues,chronic exposure to Ang II,the main effector of the RAS,increases oxidative stress, activates fibrogenesis and promotes apoptosis.An intrinsic RAS has also been demonstrated in the endocrine pancreas.ACE2 degrades Ang II and Ang I.This study intends to investigate the expression of ACE2 in human,rat and mouse exocrine and endocrine tissues of pancreas,which may give us more clue for better understanding the possible pathogenesis in pancreas lesion in SARS and diabetic patients.Methods Continuous paraffin section slides were made from human,rat and mouse pancreata for ACE2 analyses and negative controls respectively.The expressions of ACE2 protin of each of the tissues were detected by immunohistochemistry with the goat-anti ACE2 monoclonal antibody.The total RNA was extracted from rat and human pancreatic tissue(a donors tissue) by using TriZol Reagent with subsequent chloroform-isopropanol extraction according to the manufacturer's instructions,and first-strand human and rat cDNA synthesis were performed for detection of ACE2 by PCR.Results ACE2 protein was detected in whole human,rat and mouse pancreata.In the human pancreas,the distribution pattern was similar to that observed in the rat and mice: the islet showed strong positivity,and a somewhat weaker but nevertheless positive immunoreaction could be found in the acinar cells.Total RNA was extracted from rat and human pancreata tissue.The RNA samples were run down denaturing agarose gels,only samples that had intact 18S and 28S ribosomal bands were used for the study.The expression of ACE2 mRNA was detected in whole pancreata of human and rat by RT-PCR.PCR products were analyzed by agarose gel electrophoresis.The identity of PCR products was confirmed by sequence analyses.Conclusion Our data demonstrates a tissue specific pattern of ACE2 expression and suggestes that ACE2 expression in pancreas especially in islet of pancreas may cause the elevation of blood glucose levels in SARSpatients.Several large clinical trials had demonstrated that blockade of the RAS protects against the development of diabetes.The possiblemechanism is that the blockade of the RAS attenuate the deleterious actions of Ang Ⅱ on pancreatic islet structure and function and accentuate the protective actions of ACE2.So ACE2 may also play an important role in the pathogenesis of diabetes.The addition of ACE2 to the complexities of the intrinsic RAS in the endocrine pancreas may lead to the development of novel therapeutics for the treatment of diabetes.The precise role of ACE2 in pancreatic physiology and pathology will need to be studied in more detail.

Follow-up Study of Diabetic Retinopathy in Type 2 Diabetes Mellitus

Kang Lihua;Yang Jinkui;Yang Guangran;Shi Jing

2007, 28(3):  283-287. 

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Objective To investigate the incidence and progression of diabetic retinopathy in type 2 diabetes,and to analyze the risk factors associated with incidence and progression of diabetic retinopathy.Methods One hundred and sixty-four patients,diagnosed as type 2 diabetes mellitus(84 males,80 females;mean age: 58.84±9.12 years,age of onset: 50.65±9.31 years,diabetes duration: 8.21±6.73 years) from the endocrinology unit of Beijing Tongren Hospital from March 2003 to December 2004 were recruited in this study.All the patients had underwent one-field,non-mydriatic,digital photography,and had diagnosed as no diabetic retinopathy and mild or moderate nonproliferative diabetic retinopathy according to international clinical diabetic retinopathy disease severity scales in 2002.During follow-up in September 2006 to December 2006 the same method was used.According to the baseline degree of diabetic retinopathy,we divided the subjects into two groups: no diabetic retinopathy,mild or moderate nonproliferative diabetic retinopathy.The baseline age,diabetes duration,arterial blood pressure,body mass index,waist to hip ratio,fasting blood glucose,HbA1c,lipid profile were evaluated.Results After 2 years,DR developed in 22 of 104(21.2%) patients unaffected at baseline,progression of mild or moderate nonproliferative diabetic retinopathy in 22 of 60(36.7%).Logistic regression analysis showed that the duration,glycosylated hemoglobin,total cholesterol and urine albumin were independent risk factors of incidence of diabetic retinopathy;duration,body mass index and dyslipdemia were independent risk factors of progression of mild or moderate nonproliferative diabetic retinopathy in type 2 diabetes.Conclusion DR is a multifactorial microvascular complication of type 2 diabetes,which,apart from hyperglycemia,is influenced by blood pressure,lipid concentrations,urine albumin,and other factors.It is necessary to control the risk factors,with regular follow-up and in time treatment for preventing visual loss.

Construction and Expression of Multi-copied Human C-peptide Gene in Escherichia Coli

Jia Xiujuan;Yang Jinkui;Chai Sanbao;Yu Mei;Liu Xiaochao;Wei Yongxiang

2007, 28(3):  288-291. 

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Objective To obtain recombinant prokaryotic expression plasmid containing multi-copied human C-peptide in order to increase the expression of C-peptide in Escherichia coli(E.coli).Methods DNA sequence containing multiple copies of human C-peptide was obtained through introducing the restricted enzymatic site to ensure the multiple C-peptide encoding gene ligat head-to-tail.After being identified with DNA sequencing,the DNA sequence was cloned into the pET-30a expression vector.Then the recombinant expression plasmid was transformed into E.coli,induced with IPTG and purified with Ni-NTA resin to obtain the fusion C-peptide.Results A gene fragment encoding five copies of C-peptide was synthesized and cloned into the pET-30a vector.After being transformed into E.coli,induced with IPTG and purified with Ni-NTA resin,the fusion C-peptide tagged to six histidine was obtained.The fusion protein was expressed at high level as a soluble product in the cytoplasm.Ni-NTA affinity chromatography efficiently separated the expressed fusion protein from the supernatant,to obtain about 20～40mg/L of the fusion protein with 80% purity.Conclusion High-level expression of human C-peptide was obtained,which should lay an important basis for the study of the function of C-peptide.

The Survey of Relation between Overweight,Obesity,Acanthosis Nigricans and Fasting Capillary Whole Blood Glucose

Liang Xuejun;Hou Dongqing;Gong Chunxiu;Cao Bingyan;Yan Chun

2007, 28(3):  292-295. 

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Objective The strong relation between type 2 diabetes mellitus and overweight,obesity,obesity with acanthosis nigricans is widely concerned. In this study we determined the fasting capillary whole blood glucose(FCBG) of school children and adolescence and investigate the relation between overweight, obesity,acanthosis nigricans and FCBG further to make sure the normal level of FCBG for children and adolescence.Methods A total of 19 112 school children and adolescence from six regions of Beijing and aged 6 to 18 years were measured for height,weight,acanthosis nigricans and the FCBG,Their mean age was(12.564±3.280) years,They were divided into normal weight group,overweight group and obesity group according to body mass index(BMI)-overweight and obesity screening standard of Chinese school children and adolescence.Results The overall prevalence of overweight,obesity and acanthosis nigricans were 21.38%,9.7% and 2.43% respectively.In the group with FCBG≥5.6 mmol/L,the positive rate of overweight was 29.85%,significantly higher than that with FCBG <5.6 mmol/L 21.38%,(χ² Value=20.50,P<0.01).In the group FCBG ≥6.1 mmol/L,the positive rate of overweight was 35.92%, significantly higher than that with FCBG<6.1 mmol/L,21.31%(χ² Value=13.02,P<0.01).In the group with FCBG≥5.6 mmol/L,the positive rate of obesity was 15.99%,significantly higher than that with FCBG <5.6 mmol/L 9.54%(χ² Value=21.77,P<0.01).In the group with FCBG≥6.1 mmol/L,the positive rate of obesity was 20.39%,significantly higher than that with FCBG <6.1 mmol/L,9.64%(χ² Value=13.52,P<0.01).In the group of FCBG ≥5.6 mmol/L,the positive rate of obesity children and adolescence with acanthosis nigricans was 5.76%,significantly higher than that of FCBG <5.6 mmol/L2.43%,(χ² Value=22.5,P<0.01).The critical point 6.1mmol/L did not show any advantage when compared with the point 5.6 mmol/L.Conclusion 6～18 year old school children and adolescence with overweight,obesity and obesity with acanthosis nigricans correlated to FCBG high than 5.6 mmol/L in a much higher percentage.High risk children and adolescence for T2DM should get BG screening earlier.FCBG screening as early as possible would help prevent children and adolescence with overwight obesity and obesity with acanthosis nigricans from getting type 2 diabetes mellitus,especially for those who can not accept oral glucose tolerance test.We suggest the critical point of FCBG as 5.6 mmol/L.

Studies on the Inter-relationship between the Level of Serum TNF-α and Type 2 Diabetes Mellitus Complicating Metabolic Syndrome

Lun Yuanyuan;Wang Li;Yang Jinkui

2007, 28(3):  296-299. 

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Objective To investigate the role of serum tumor necrosis factor-α(TNF-α) in the pathogenesis of type 2 diabetes mellitus complicating metabolic syndrome.Methods Two hundred and fifty seven patients were diagnosed as T2DM,they were found to have acute or chronic infections,as well as other endocrine diseases,their liver function was normal.According to the 2005 IDF MS definition,the patients were divided into MS(T2DM with MS,n=174) and NMS(T2DM without MS,n=83),control group included 43 healthy subjects.The serum levels of TNF-α with RIA.Differences in blood lipid,HbA1c CRP,PAI-1,IL-1,IL-2,IL-6,IL-8 and so on among groups were observed.Data were analyzed with SPSS 11.5.Results ① The TNF-α levels in T2DM patients with MS were significantly higher compared with patients without MS,and the TNF-α levels in patients with T2DM were higher than those in the control group(P<0.01,P<0.05) respectively.② The incidence of macro-vascular complications in group MS was 60.9%,and in group NMS 45.8%,the difference was significant statistically(P<0.05).The incidence of fatty liver in group MS was 39.1%,and in group NMS 15.7%(P<0.01).③ By variable correlations showed a positive correlations between the serum TNF-α level and TG,TCH,FFA,FCP,IL-1,IL-2,IL-6 and IL-8.Multiple regression showed WHR,TG,LDL-C,IL-1,IL-2 and IL-6 were equal predictors of serum TNF-α level.Conclusion The level of serum TNF-α in patients with T2DM is higher compared with the control group.The level of serum TNF-α in the T2DM patients with MS is higher than the T2DM patients without MS.This indicates serum TNF-α plays an important role in T2DM,and it may facilitate the development of MS.Multiple regression showed WHR,TG,LDL-C,IL-1,IL-2,IL-6 and IL8 are equal predictors of serum TNF-α level.In the group T2DM with MS,the morbidity of macro-vascular complication and fatty liver is significantly increased compared with that in the group T2DM without MS,this indicates that disorders of metabolism and inflammationplay an important role in the development of complications of T2DM.The aim of treating T2DM therefore not only lowering blood glucose,but also controlling the level of blood lipid and inflammation.

The Relationship between Skin Micro-circulation of Lower Extremity and the Diabetic Duration in Type 2 Diabetes

Liu Xiaoyan;Fu Hanjing;Feng Yajuan

2007, 28(3):  300-303. 

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Objective To investigate the relationship between the skin micro-circulation changes and disease duration in type 2 diabetic patients.Methods Micro-circulation of lower extremity in 50 type 2 diabetic patients whose disease durations were less than 5 years(n=24) and more than 15 years(n=26),and age-sex-matched control subjects(n=30) were measured with laser doppler flowmeter(LDF) and by the method of post occlusive reactive hyperemia(PORH).The first step was to measure rest flux of the skin of the left foot(PORH_rest),then the lower extremity flow was occluded at higher than 20 mmHg suprasystolic pressure higher than for 2 min.Skin blood flux was measured during and after occlusion using LDF continuously.PORH parameters were rest flux of the skin(PORH_rest),maximum increase in hyperemia perfusion(PORH_max),time-to-peak(T_p),amplitude of peak perfusion(PORH_peak),PORH_max=PORH_peak-PORH_rest and PORH_max/T_p=(PORH_peak-PORH_rest)/T_p.All diabetic patients were given tests for fasting plasma glucose(FPG),serum creatinine,blood routine,glomerular filtration rate(GFR),urinary albumin excretion rate(UAER),left lower extremity blood pressure,calculated body mess index(BMI) and waist-to-hip ratio(WHR).Results 1) The longer diabetic duration,the lower the PORH_peak of skin micro-blood-flow in diabetes and less than controls(controls 21.67±3.15,diabetic duration less than 5 years group 16.61±6.67,diabetic duration more than 15 years group 10.54±3.22,P<0.01).2) PORH_max of skin micro-blood-flow in diabetes at lower extremity skin was significantly reduced in Type 2 diabetic patients compared with the controls(controls 12.87±3.68,diabetic duration less than 5 years group 9.76±5.89,diabetic duration more than 15 years group 5.08±3.14,P<0.01).3) Bivariate correlations analysis showed and that the correlation factors of PORH_peak from big to small were: diabetic duration(r=-0.538,P<0.01),log urinary albumin excretion rate(UAER)(r=-0.447,P<0.01),glomerular filtration rate(GFR)(r=0.368,P<0.05),age(r=-0.365,P<0.05).The correlation factors of PORH_max from big to small were: diabetic duration(r=-0.497,P<0.01),log urinary albumin excretion rate(UAER)(r=-0.480,P<0.01),glomerular filtration rate(GFR)(r=0.318,P<0.05),FPG(r=-0.301,P<0.05).Linear regression analyze showed that the relationship of PORH_peak and diabetic duration was the most intimate(the standardized coefficients of diabetic duration was-0.575,P< 0.01);The relationship of PORH_max and diabetic duration was also the most intimate(the standardized coefficients of diabetic duration was-0.533,P<0.01).4) Regrouping the type 2 diabetic patients(n=50) by HbA1c(HbA1c<10%,n=26;HbA1c ≥10%,n=24),age,sex,logUAER of two groups were not statistically significant(P>0.05),PORH_peak and PORH_max of two groups were also not statistically significant(P>0.05).Conclusion The longer the diabetic duration,the worse skin micro-circulation of lower extremity in type 2 diabetes.

Effects of Leptin on Gene and Protein Expression and Tyrosine Phosphorylation of Leptin Receptor in Rat hepatocyte

Guo Qiyu;Gao Yan;Cong Lin;Shao Ningsheng

2007, 28(3):  304-306. 

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Objective To investigate effects of leptin on gene and protein expression and tyrosine phosphorylation of leptin receptor in rat hepatocytes.Methods hepatocytes were isolated from new-born Sprague-Dawley rats,after incubation with leptin(5 nmol/L) for 12,24 and 36 h,Western bolt was used to assess the protein level of leptin-receptor.The mRNA expression level of leptin-receptor was assayed by RT-PCR.The tyrosine phosphorylation of leptin-receptor weas assayed by immuno-precipitation.Results After incubation with leptin(5 nmol/L) for 12,24 and 48 h,there was no difference in the protein contents of leptin receptors between leptin(5 nmol/L) treated and control hepatocytes(P>0.05).The mRNA content of leptin receptor in rat hepatocytes was similar to that of the control after incubation with leptin(5 nmol/L) for 12 h(P>0.05),but significantly decreased after 24 or 36 h(24 h: leptin(5 nmol/L) 0.37±0.05 vs control 0.33±0.03,P<0.05;;36 h: leptin(5 nmol/L) 0.36±0.04 vs control 0.32±0.01,P<0.05).The tyrosine phosphorylation of leptin-receptor was remarkably reduced after incubation with leptin(5 nmol/L) for 24 or 36 h(24 h: leptin(5 nmol/L) 0.30±0.05 vs control 0.27±0.05,P<0.05;36 h: leptin(5 nmol/L) 0.32±0.05 vs control 0.26±0.07,P<0.05).Conclusion Leptin(5 nmol/L) may up-regulate leptin receptor gene expression and its tyrosine phosphorylation in rat hepatocytes and may contribute to promoting the signal transduction of leptin itself.

Analysis of Risk Factors of Peripheral Artery Disease in Elder Type 2 Diabetic Patients

Fan Hong;Zhong Liyong

2007, 28(3):  307-309. 

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Objective To analyze the risk factors of peripheral artery disease in elder diabetic patients.Methods 240 elder type 2 diabetic patients were enrolled.Patients were divided into three groups.27 patients had not peripheral artery disease;154 had moderate peripheral artery disease;59 had severe peripheral artery diseases.The relationship between seventy of peripheral artery diseases judged by doppler ultrasonic and biochemical parameters was investigated.Results In the moderate peripheral artery disease group,peripheral artery diseases were correlated to the retinopathy.In the severe peripheral artery disease group,peripheral artery diseases were correlated to family history、TC、LDL-C、pathological changes of perpheral nerve and retinopathy.Conclusion Peripheral artery diseases in the elderly diabetic patients correlate with the course of DM and LDL-C.

Effects of Endotoxemia on Oxidative Stress and NO Levels of Streptozocin-induced Diabetic Rats

Yang Zhihong;Zhong Liyong;Wang Hui

2007, 28(3):  310-312. 

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Objective To investigate the effects of endotoxemia on parameters of oxidative stress and NO levels of streptozocin-induced diabetic rats.Methods Experimental diabetic animal models were constructed by streptozocin injection(50 mg/kg,i.p).The endotoxemia condition was then induced by LPS(0.5 mg/kg,i.p.).Parameters related to oxidative stress such as SOD,MDA,GSH,NO and concentrations of glucose and insulin were measured at 6,12,24 and 72 h after injection of LPS.Results The levels of serum NO and MDA elevated significantly at 24 h after LPS injection(P<0.05),whereas the levels of SOD and GSH decreased significantly at 24 h after LPS injection(P<0.05).Though no significant difference was seen in blood glucose levels in these DM groups before LPS injection,they elevated at 12,24 and 72 h after LPS injection.The insulin levels decreased in diabetic rats significantly at 24 and 72 h after LPS injection.Conclusion Endotoxemia would reduce the antioxygenic ability in diabetic rats,elevate the production of NO and impair the regulatory ability to blood glucose.

基础研究

The Effect of Hypoxia on the Proliferation and Differentiation of Adult Neural Stem Cells in Situ

Wang Xuan;Wu Haitao;Zhu Lingling;Fan Ming;Wang Xiaomin

2007, 28(3):  313-317. 

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Objective Many experiment results have confirmed that neural stem cells(NSCs) exist in adult central nervous system(CNS).These NSCs will be activated to proliferate,migrate and differentiate by some factors such as growth factors,neurotrophic factors,injury,glucocorticosteroids,exercises,and so on.Intermittent hypoxia is a lesion factor suffered with frequently.Some experiments in vitro have showed that intermittent hypoxia increased the proliferation capacity of NSCs.In present study,the effect of intermittent hypoxia on the proliferation and differentiation of in situ adult NSCs in 6-hydroxydopamine(6-OHDA) lesioned rat brains was investigated.Moreover,the behavioral change was also observed.Methods 6-OHDA was injected into one-side medial forebrain bundle(MFB) of adult rats to establish a model of Parkinson's disease(PD).Before treated with hypoxia exposure,successful parkinsonian model rats were intraperitoneally injected with 5-bromodeoxyuridine(BrdU,100 mg/kg weight) and then treated with 3000 meters high altitude 4 h/d for consecutively 2 weeks.After that,these animals were placed back into normoxia environment for another 2 weeks.Animal behavior test and histology experiments were carried out after two-weeks hypoxia and two-weeks normoxia exposure.The mRNA expression of hypoxia induction factor(HIF)-1α in striatum near to subventricular zone(SVZ) and hippocampus was examined by RT-PCR array.Results Hypoxia did not affect the behavioral disorders of parkinsonian rats.Immunostaining results showed that under hypoxia condition,the number of newborn cells increased greatly,especially in SVZ.Neonatal cells were not found in substantia nigra and possessed the very low differentiation capacity,and they were not found to give rise to neurons and astrocytes.In striatum lesioned by 6-OHDA injection,the neonatal cells did not undergo committed differentiation procedure to become tyrosine hydroxylase(TH)-immunoreative dopaminergic neurons.Contrary to control,the expression of HIF-1α in striatum and hippocampus was not influenced by hypoxia exposure.Conclusion The results showed that intermittent hypoxia could activate the proliferation of adult NSCs in situ,however,it could not induce these neonatal cells to differentiate into specific dopaminergic neurons which was the target neurons in the therapy for PD.These primary results indicated that simple injury factor such as 6-OHDA or hypoxia,could not contribute to thecommitted differentiation,and proliferating NSCs could not restore the function lesion of nervous system.Combination with other inductionfactors,for example,neurotrophic factors,may help the certain differentiation of adult NSCs in situ,thereby benefit the therapy of neurodegenerative diseases.

Studies on Neural Differentiation of Adult Rat Skeletal Muscle-derived Stem Cells in Vitro

Yang Jian;Jia Jun;Wang Xuan;Wang Xiaomin

2007, 28(3):  318-323. 

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Objective Skeletal muscle-derived stem cells are a population of cells including several different kinds of immature progenitors residing in the skeletal muscle tissues,such as pluripotent stem cells,side population cells,satellite cells and activated satellite cells-myoblasts.These precursor cells function in the maintenance and repair of adult skeletal muscle injury and regeneration.It has been demonstrated that mice myogenic cell lineage C2C12 can be induced to differentiate into neurogenic lineage by genetic modification.One kind of muscle-derived stem cells,myoblasts derived from adult rat skeletal muscles,has not been known to possess this neural conversion ability.As a type of cell population,muscle-derived stem cells can provide the sufficient cell source,which will benefit not only the muscular atrophy,but also the central nervous system neurodegenerative diseases such as Parkinson's disease.Because these cells can be obtained from the patient per se,they possess the advantage of clinical value.Therefore,in this study,the feasibility of neural differentiation of adult rat skeletal muscle-derived stem cells in vitro has been investigated.Methods Tissue was isolated from six-months old Sprague-Dawley rat gastrocnemius muscle.Cells were enzymatically dissociated by the addition of a solution of dispase,collagenase,and CaCl₂.The slurry,maintained at 37 ℃ for 45 min,was triturated every 15 min,and then passed through 75 μm nylon mesh.The filtrate was centrifugalized at 1 000 r/min to precipitate the dissociated cells,the pellet was resuspended in F10 medium,and the suspension was plated on type I collagen-coated dishes.During the first 4 days of the primary cultures,cells were enriched by continuous differential attachment technique.After culturing for 4～6 passages,cells were plated as a suspension in a medium optimized for neural stem cells growth which contained Neurobasal A supplemented with B27,basic fibroblast growth factor and epidermal growth factor for 7～10 d.The expression of myoblast-specific protein desmin,neural stem cell-specific protein nestin and neuron-specific protein βⅢ-tubulin、microtubule associated protein 2 were detected with immunocytochemical techniques,western blot and realtime-PCR.Results Stem cells isolated from adult skeletal muscle tissues proliferated rapidly after 4 passages,and survived for 6 months in vitro.The sequential adhesion method threw away many fibroblast cells mixed in the precursor cells culture system.After proliferation andpurification,over 95% of the muscle-derived stem cells expressed desmin and some of them expressed βⅢ-tubulin,but none of these cells expressed nestin or microtubule associated protein 2.After 7～10 d cultivation in the medium optimized for neural stem cells growth,muscle-derived stem cells formed neurosphere-like aggregates.Identified by immunocytochemical methods,western blot and realtime-PCR,expression of nestin increased significantly and that of desmin declined sharply.βⅢ-tubulin expression was slightly reduced and microtubule associated protein 2 expression was not detected after induction.Conclusion Muscle-derived stem cells were highly enriched through our isolation and cultivation system,and 95% were desmin-positive myoblasts.Through replacement of F10 medium with medium optimized for neural stem cells growth,muscle-derived stem cells could be induced to differentiate into neural stem cells after 7～10 d in vitro.The expression of myogenic markers declined gradually and that of neurogenic markers increased.This indicates that in our present culture system,muscle-derived stem cells could be induced to process the neural conversion progression.Next step,we will induce the primary adult stem cells derived from skeletal muscle tissues to differentiate into neurons by some factors,in order that the muscle-derived stem cells can be used as a kind of cell source for nervous system diseases especially neurodegenerative diseases treatment.

Coexistence of AD7c-NTP and Insulin Receptor in CA1 Zone of Hippocampus in Mouse

Wang Hongjuan;Zhao Zhiwei;Ji Zhijuan;Sheng Shuli;Wang Rong

2007, 28(3):  324-327. 

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Objective To investigate the distribution of Alzheimer-associated neuronal thread protein(AD7c-NTP) and the coexistence of AD7c-NTP and insulin receptor in CA1 zone of hippocampus in mouse.Methods SDS-PAGE and Western blot were applied to assess the molecular weight of AD7c-NTP in the extract of protein in the brain of mouse and the specificity of the antibody of AD7c-NTP.The distribution of AD7c-NTP in CA1 zone of hippocampus in mouse was investigated with immunohistochemical methods.The coexistence of AD7c-NTP and insulin receptor was studied with double immunohistochemical staining.Results A band of about 41 000 was detected with the antibody of AD7c-NTP,which was coincided with the previous report.AD7c-NTP immunoreactive positive cells were observed in CA1 zone of hippocampus in mouse,whose cellular membrane and cytoplasm were stained.AD7c-NTP and insulin receptor were found coexisting in the same cells in CA1 zone of hippocampus.Conclusion AD7c-NTP is found in CA1 zone of hippocampus in mouse,and coexists with insulin receptor.It indicates that AD7c-NTP may have relation to with the insulin signal pathway.

Presence of α-Synuclein in Rat Brain and Its Ability to Transport into the Mitochondria

Liu Guangwei;Yin Juanjuan;Zhang Chunyan;Li Xin;Liu Yaobo;Li Yaohua;Yang Hui;Chen Biao;Yu Shun;

2007, 28(3):  328-332. 

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Objective The abnormality in α-synuclein(α-SYN) expression and the deficiency in mitochondrial function are both implicated in the pathogenesis of Parkinson's disease(PD).And many evidence strongly support a linkage between α-SYN and mitochondria in the pathophysiology of PD,but the molecular basis for the linkage is unknown.Our work is to study whether α-SYN is present in normal rat brain and is able to be transported into mitochondria to help analyzing the asociation of α-SYN with mitochondria in the pathogenesis of PD.Methods Western blot analysis was performed on the isolated brain mitochondria using different anti-α-SYN antibodies(3D5,2E3,EQV1,MDV2,and PQE3),which identify different parts of αSYN.To make clear the precise localization of α-SYN in the mitochondrion,pre-embedding immunoelectron microscopy was performed using 3D5 monoclonal antibody to probe mitochondrial α-SYN and the second antibody-conjugated ultra-small golds to label the 3D5 primary antibody.The total cell α-SYN has been shown to be expressed unevenly in the brain,with its amount being much greater in olfactory bulb,hippocampus,striatum,thalamus and cerebral cortex than in other regions.To answer whether the mitochondrial α-SYN is also richer in these brain regions,mitochondria from different brain regions were isolated,and the relative amount of α-SYN was determined using Western blot analysis.Different concentrations of recombinant human α-SYN was incubated with isolated cerebellar mitochondria where there was no endogenous α-SYN.The reaction mixture was incubated at 37 ℃ for different time.And after thoroughly washing the free α-SYN western blot analysis was used to examine the transportation of α-SYN into mitochondria.Digitonin was used to disrupt the outer mitochondrial membrane to rule out the possibility that the α-SYN transport into mitochondria is because of non-specific binding to the outer mitochondrial membrane.Results Immunoblot analysis identified αthe presence of α-SYN in normal rat brain mitochondria.And shown by immunoelectron microscopy,except sparsely distributed gold particles in the cytoplasm of neurons,densely distributed gold particles are observed in the mitochondria,where the gold particles are localized in the cleft between outer and inner membranes and mitochondrial matrix inside the inner membrane.Mitochondrial α-SYN is differently expressed in different brain regions.In accordance with the total cell α-SYN,the levels ofmitochondrial α-SYN were also higher in olfactory bulb,hippocampus,striatum and thalamus than in cerebral cortex,brain stem and cerebellum.The sequence of the amount of α-SYN in different regions was striatum > hippocampus> olfactory >bulb >thalamus >cortex >brainstem >cerebellum.Incubation of α-SYN protein with isolated mitochondria led to transportation of this protein into the mitochondria in a dose-dependent manner.The import of α-SYN into mitochondria was rapid but not time-dependent manner,with the earliest detection of α-SYN protein in the mitochondria within 5 min.After disrupting the outer membrane with digitonin,most of α-SYN remained in the inner membrane fraction.This suggested that α-SYN is transported into mitochondria rather than bound to the surface of the outer membrane.Conclusion α-SYN is normally present in rat brain mitochondria but varied in expression levels in different brain regions with the amount of α-SYN being much greater in olfactory bulb,hippocampus,striatum and thalamus than in cerebral cortex,brain stem and cerebellum.Except cerebral cortex and cerebellum,the levels of mitochondrial α-SYN is in accordance with the total cell α-SYN,raising a possibility that the αSYN in mitochondria may come from the cytoplasm.Our data provide new evidence for the linkage between α-SYN and mitochondria.Thorough discovery of the role of α-SYN in mitochondria may shed a light on the pathological role of α-SYN in PD.

Preparation and Identification of Recombinant Rat α-Synuclein

Li Xin;Yu Shun;Li Yaohua

2007, 28(3):  333-336. 

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Objective To investigate bacteria expression of rat α-Synuclein cDNA,and prepare the recombinant protein.Methods Total RNA were isolated from Wistar Rat brain.A cDNA fragment encoding the α-Synuclein was amplified by RT-PCR,and was subcloned into plasmid pGEX-4T-1.The recombinant plasmid DNA pGEX-raSYN was expressed with bacteria BL21(DE3),and Glutathione S transferase(GST) tagged protein was purified by column chromatography.Recombinant rat α-Synuclein was freed by thrombin cleavage,and confirmed with SDS-PAGE and western blot analysis.Results Amplified cDNA fragment was composed of 420 bp and encodes 140 amino acids.The fusion protein GST-raSYN was expressed in the bacteria,with IPTG present in the condition medium.The recombinant rat α-Synuclein was purified to homogeneity from soluble fraction of the bacteria.The molecular weight of the purified protein was calculated to be approximately 524 000 on superdex S-200HR and to be 18 000 on SDS-PAGE in the presence of b-mercaptoethanol.Western blot analysis demonstrates that the purified recombinant protein was reactive to the anti α-Synuclein monoclonal or polyclonal antibodes.Conclusion Rat α-Synuclein cDNA was effectively expressed with bacteria,and pure recombinant rat α-Synuclein was prepared,which may be samples to studies physiological function of the protein.

The Effects of CoCl₂ Pretreatment on HIF-1α Expression in Hippocampus of Hypoxic Preconditioned Mice

Shao Guo;Gao Cuiying;Lü Guowei;

2007, 28(3):  337-340. 

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Objective To investigate the effect of CoCl₂ pretreatment on expression of HIF-1α in the hippocampus of hypoxic preconditioned mice.Methods Balb/c mice were randomly divided into group of CoCl₂ and normal saline(NS),respectively injected with CoCl₂ and NS 3 h before exposure to hypoxia for 0 run(H0),1 run(H1) and 4 runs(H4).The tissue of hippocampus was removed immediately after the exposure.Technique of Western blot and electrophoretic mobility shift assay(EMSA) were used to measure the HIF-1α protein level and HIF-1α DNA binding activity,respectively.Results The hypoxic tolerance time in group NS was significantly increased run by run but unchanged in group CoCl₂.In group CoCl₂,the level of HIF-1α protein was unchanged whereas HIF-1α DNA binding activity was increased in H1 and decreased in H4.In group NS,both HIF-1α protein and DNA binding activity were increased.Conclusion Hypoxic-tolerance of mice to hypoxia and HIF-1α in their hippocampus are,respectively,increased and activated by CoCl₂ while its underlying molecular mechanisms are thought to be different from those induced by acute repetitive hypoxic exposure.

Mechanosensitivity of ATP-Sensitive Potassium Channels in Rat Colonic Smooth Muscle Cells

Wang Wei;Huang Haixia;Zhang Lijuan;Ma Tao;Liu Ping;Wei Hua;Niu Weizhen

2007, 28(3):  341-344. 

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Objective To study the expression and mechanosensitivity of ATP-sensitive potassium channel(K_ATP) in rat colonic smooth muscle cells.Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to measure mRNA expression of K_ATP in colonic smooth muscle.K_ATP channel activities were investigated with patch-clamp techniques and pressure-clamp techniques in inside-out mode on isolated smooth muscle myocytes.Results RT-PCR amplified the expected product of 727 bp for K_ATP from total RNA isolated from colonic smooth muscle.In this mode,K_ATP activities were recorded on isolated smooth muscle cells,the mean conductance was(44.5±1.5)pS at +60 mV.The activities were blocked by glibenclamide(10 μmol).The open probability of K_ATP channels was increased by-40 mmHg suction pressure.Conclusion K_ATP channels are expressed in rat colonic smooth muscle cells and are sensitive to stretch stimuli.These results suggest that the channel may be involved in the regulation of electric and mechanic activities of colonic smooth muscle in rats.

Transportation of Functional Fragments of α-Synuclein into Dopaminergic Neuronal Cells and Their Effects on Proliferation of the Cells

Yin Juanjuan;Han Junyan;Li Xin;Li Yaohua;Liu Yaobo;Yu Shun;

2007, 28(3):  345-349. 

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Objective α-Synuclein(α-Syn) is a 140 amino acid protein, which is strongly implicated in the pathogenesis of Parkinson's disease.The amino acid sequence of this protein can be divided into three distinct domains: a highly conserved amino terminal(residues 1～60) with four KTKEGV repeats,which mediates its binding to phospholipid membranes;A central hydrophobic NAC domain(residues 61～95) with two additional KTKEGV repeats,which makes up the highly amyloidogenic part of the molecule,and a carboxyl terminal acidic tail(residues 96～140),which is composed primarily of glutamate and aspartate residues that appears critical for the chaperone-like activity of α-Syn.Previous studies have shown that this protein can be secreted from the cells and detected in the culture medium and cerebrospinal fluid.Whether or not the extracellular α-Syn may affect neuronal function remains unknown.We have previously reported that α-Syn,when applied extracellularly,can enter into MES 23.5 dopaminergic cells and promote proliferation of the cells.However,β-Syn,another member of synuclein family,which is highly identical to α-Syn in N-terminal but different from α-Syn in NAC domain and C-terminal,showed no effect on cell proliferation.These findings indicate that the effect of α-Syn on cell proliferation is highly dependent on its molecular structure.In this study,we will study the transportation of three functional fragments of α-Syn into dopaminergic neuronal cells and their effects on the proliferation of the cells.Methods Human recombinant α-Syn and its three functional fragments: N-terminal(α-Syn 1～65 amino acid),NAC(α-Syn 60～95 amino acid) and C-terminal(α-Syn 96～140 amino acid) were expressed in E.coli by a prokaryotic expression system.The proteins expressed were then passed through a column of Glutathione Sepharose 4B,purified by gel filtration and reverse chromatograph,respectivly.The correction of the full-length α-Syn and its three functional fragments were identified by Western blot analysis using specific antibodies against different parts of α-Syn.The full-length α-Syn and its three functional fragments were then added to the culture medium of MES 23.5 dopaminergic neuronal cells.The entry and subcellular localization of the proteins were identified using immunofluorecent labeling.The effect of the proteins on cellproliferation was measured with cell growth curve drawn by cell counting.Results Western blot analysis using specific antibodies againstdifferent parts of α-Syn demonstrated the correction of the full-length α-Syn and its three functional fragments.After applied to the culture medium,the full-length α-Syn and its three functional fragments were shown to enter into the cells.However,their subcellular localization in the cell was different.The full-length α-Syn was distributed in both cytoplasm and nucleus with its amount higher in the nucleus than in the cytoplasm.The N-terminal fragment was presented in the cytoplasm and processes of the cells.The NAC fragment was only seen in the cytoplasm. The C-terminal fragment was mainly observed in the nucleus.Growth curve showed that the full-length α-Syn and its NAC and C-terminal fragments promoted cell proliferation.In contrast,the N-terminal fragment had no effect on cell proliferation.Conclusion The extracellularly applied recombinant α-Syn and its three functional fragments can enter into MES23.5 dopaminergic cells with different subcellular localizations.The promotion effect of α-Syn on cell proliferation may depend on its NAC and C-terminal amino acid sequences.The full-length α-Syn and its NAC and C-terminal fragments promoted cell proliferation,while the N-terminal fragment had no effect on cell proliferation.These results suggest that the NAC and C-terminal of α-Syn may be responsible for its role in cell proliferation.

Rats Model of Abdominal Aortic Banding and Exercise Test on Evaluation of Cardiac Function

Ma Li;Liu Jie;Chu Nan;Shi Xubo;Yang Jingang;Li Tianchang;Hu Dayi

2007, 28(3):  350-354. 

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Objective To establish rat model of chronic heart failure by partially banding abdominal aorta for different time limits and to elucidate the clinical implication of exercise test on evaluation of cardiac function of the rat model of chronic heart failure.Methods Eighty Wistar rats were randomly divided into two groups: aortic banding group and sham-group(each containing 40 rats).Cardiac mass index,exercise parameters,hemodynamic parameters and cardiac morphological changes were investigated at different time(4,8,12 and 16 weeks after the operation) in different groups.Results Left ventricular end-diastolic pressure(LVEDP) increased greatly in aortic banding group compared with sham-groups(after 8 weeks,P<0.05).The diastolic parameter of-dp/dt decreased in 4 weeks after aortic banding(P=0.054),and lower than sham-group significantly in 8 weeks after operation(P<0.001).Left ventricular systolic pressure(LVSP) and +dp/dt increased after 12 weeks of aortic banding operation(P<0.05) compared with sham-group.Cardiac mass and cardiac mass index increased from 4 weeks after aortic banding(P>0.05) compared with sham-group,and more significant higher than the sham-group in 8 weeks after operation(P<0.05).Cardiac myocardial diameter(CMD) increased compared with sham-group from 4 weeks after aortic banding(P<0.001).Parameters of exercise test in aortic banding group decreased compared with sham-groups of corresponding time points(P>0.05),and more significant in 16 weeks after operation(P<0.01).And the parameters of exercise test in aortic banding group of 16 weeks after the operation were also lower than the baseline(P<0.001).Conclusion Abdominal aortic banding can successfully copy chronic heart failure model in rats.Exercise test can be successfully used for the evaluation of cardiac function of the model.

A Worm Model for Automatic Segmentation of Medical Image

Feng Jian;Luo Shuqian

2007, 28(3):  355-358. 

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Objective The correct classification of human organs in medical image can provide computer-assisted diagnosis for clinicians,and also it is the groundwork of some image processing,such as the 3D reconstruction and the visualization of medical images.There are many segmentation algorithms by far,such as threshold segmentation method,differential coefficient operator edge detection method,region growing method and clustering segmentation method,however,they are far from perfect.Most of them are semiautomatic algorithms which can not receive satisfied segmentation result without the intervention of experts.Medical image segmentation has always been the research hotspot in the medical image analysis area,and is also the key step to finish the quantitive analysis of the anatomical structure and extract the related diagnosis information.The variety properties of image as well as the mixture with a lot of noise lead to the main difficulties in the image segmentation.In this article we try to increase the accuracy of segmentation by applying the principle of artificial life to automatic segmentation of images.Methods The method based on artificial life is useful in understanding biological rules,and it also has been successfully applied in the robot and computer graphics.It is a new trial to apply the principle of artificial life to medical image segmentation.We constructed an intelligent deformable model called worm model with the application of image segmentation based on artificial life and created a worm model.The worm model,no larger than several pixels, is made up of many nodes,and it possesses 'head','neck' and 'body'.The 'head' is the function section,which has a nerve center system,vision system and perception system.It also can memorize information and give off orders.The 'neck' and the 'body' are locomotion parts.The worm has a central nervous system,vision system,perception system and locomotion system.It can memorize,recognize objects and control the motion of its body.Firstly we converted a MR image into a binary image,and then a worm model was placed on the top of this image.Using the prior experience,the worm would automatically find the corpus callosum.Then it would go down to the CC's center where it would divide into four little worms in order to detect edges in different directions.We commanded these worms detect image edges in four different directions: up-left,up-right,bottom-left,and bottom-right.At last,the detecting tracks were memorized when the image edge was detected completely.Results The worm model overcomes the defects of the current existing methods since it is able to intelligently process the segmentation of the image using more available information,rather than only using pixels and gradients.Its ability to segment the CC from MRI brain images showed that the worm model could segment images automatically and accurately.For those images that are more complex or with fragmentary boundary,the predominance of the worm model is especially clear.Conclusion The model is able to intelligently process the segmentation of images using more comprehensive information rather than using pixels and gradients only.The worm's biological activities are analyzed deeply and it is proved that the worm model can be used in image segmentation.This model has nerve center system,vision system,perception system and motor system,and it also has some life characteristic,such as memorizing,cognizing and locomotion controlling.Because the model has alterable eyeshot,as well as can deformflexibly,it is able to take full advantage of local information.As a result,it can automatically segment the target using more informationthan just pixels and edges,and then it can overcome the defects of the traditional algorithms.Further more,it has a high intelligence and only need a few prior experiences.The experiment result based on simulation data and real data proved the validity of the method.

Multiple Fractal Analysis on the Research of Normal Appearing White Matter in Patients with Multiple Sclerosis

Nie Shujun;Tong Zhongyong;Yu Chunshui;Tong Longzheng

2007, 28(3):  359-362. 

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Objective To discriminate the differences between normal appearing white matter(NAWM) in the patients with multiple sclerosis(MS) and normal white matter(NWM) in the healthy,and to set up a model to discriminate two classes.Methods The total sample set was 120 images with NAWM and NWM of 60 each.The NAWM sample set of 60 was randomly divided into training section and testing section by rate of 70% and 30% and the NWM was processed with the same way.Multiple fractal analysis was used to analyze regions of interest(ROI) which were gained from T2-weighted MR images of patients with MS and the healthy.Three eigenvectors were obtained,which were fractal box dimensions for 2D,3D surface and 3D volume.The sample was conversed to binary image firstly and different patterns of square were used to cover it.A data counting the covering squares was obtained from one pattern and a data set was achieved from all patterns.After logarithms of the square sizes and the data set gained were extracted,the linear fitting was conducted to calculate slope by these logarithms,i.e.fractal box dimension for 2D.Similarly to the fractal box dimension for 2D,a curved face was constructed based on the grey value of every pixel firstly,and then a 3D surface was achieved following by covering the face with different sizes of cube.After logarithms were applied to the cube sizes and the data set gained,linear fitting slope was acquired,i.e.fractal box dimension for 3D surface.When the whole volume was covered by different sizes of cube,fractal box dimension for 3D volume was then achieved.T-test was conducted to test if there were any significant differences in three eigenvectors of fractal box dimensions above between two groups of NAWM and NWM.The probabilistic neural network(PNN) was used to classify ROI based on significant eigenvectors.Results In two groups of NAWM and NWM,the mean values were 1.803 vs 1.833 for fractal box dimension for 2D(P=0.037),1.9751 vs 2.058 for fractal box dimension for 3D surface(P<0.001),2.6988 vs 2.7649 for fractal box dimension for 3D volume(P<0.001) respectively.The identification rates were 77.5% for NAWM and 65% for NWM based on these ROI.The rate of classification with fractal box dimension was close to that of other methods.Conclusion There were significant differences of fractal box dimensions between groups of NAWM and NWM,which indicated that the textures in MRI images of them could be different in a certain extent.Some microstructures change because of demyelination around nerve axes in NAWM,which perhaps leads to fractal box dimensions from NAWM a little less than those from NWM.But the pathological changes in NAWM were not macroscopic.Fractal dimension maybe becomes an effective tool used to observe images with MS of MRI.The result was affected by some factors,such asanatomization structure and the course of diseases.The data of our patients is limited,and bigger crowd is expected to confirm theconclusion above in further study.

The Inhibitory Component of the Compound Postsynaptic Potentials in the Ventral Dendrite of Crucian Carp's Mauthner Cell Evoked by Skin Stimulation

Hu Zhengjuan;Zhang Lijuan;Tong Xuehong;Yu Fen;Li Xiaoyi

2007, 28(3):  363-366. 

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Objective To explore the inhibitory component of the compound postsynaptic potentials(PSPs) in Mauthner cell's ventral dendrite.Methods Direct stimulation the skin of crucian carp,change the stimulus strength and the stimulus frequency or inject hyperpolarizing currents to the ventral dendrite through the Cl^- filled microelectrode or drop strychnine to the surface of medulla,microelectrode were used to record the change of postsynaptic potentials in the ventral dendrite.Results The PSP amplitudes were stimulus strength-and frequency-and Cl^- dependent within some limits.After dripping strychnine on the brain of crucian carp,the PSP amplitudes enhance greatly,even action potentials can break out.Conclusion It suggests that the afferent pathways from skin to M-cells contain both excited and inhibitory components,the inhibitory neurons project not only to the soma but also to the ventral dendrite,and the neurotransmitter released may be glycine.

The Investigation about the Reaction of Naturally Occurring Alkaloid Quaternary Ammonium Salts

Zhang Jianwei

2007, 28(3):  367-369. 

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Objective To investigated the reaction situation of quaternary ammonium salts with base.Methods Atropine reacted with bromoacetonitrile to give N-cyanomethyl-atropine bromide which was treated with sodium borohydride.In order to simplify the study,hydrolysis of atropine afforded tropine.Oxidation of tropine with chromic oxide gave tropinone which can be quaternized to 5 in quantitative yield.Quaternary ammonium salts 5 was treated with NaH.Results N-cyanomethyl-atropine bromide was treated with sodium borohydride to give 3 and 4.Reaction of N-cyanomethyl-tropinone bromide with sodium borohydride leads to 7.Conclusion Atropine derivatives react with sodium borohydride to performed Hofmann elimination reaction and cyclize to produce 7.

临床研究

The Study on Incidence of Depression and Correlated Factors in Patients with Parkinson's Disease

Liu Hongjun;Zhao Lijie;Ding Hui;Chen Biao

2007, 28(3):  370-372. 

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Objective To determine the incidence of depression and correlated factors in Parkinson's patients.Methods 117 Parkinson's patients were diagnosed and assessed using Self-rating Depression Scale(SDS) and Hamilton Depression Rating Scale(HAMD).The incidence,disease severity and correlated factors of depression were statistically analyzed,and a factor analysis was carried out on HAMD.Results The incidence of depression in Parkinson's patients was 43.6%,of which 88.2% were mild depression.Univariate analysis revealed that a high frequency of depression occurred in patients with bad economic status,long Parkinson's disease(PD) duration,high H&Y stage and Webster score.Depression was found to be only significantly correlated to disease status(the comparisons of Webster score had statistically significance) by means of Logistic regression analysis.According to each factor analysis of HAMD,the scores of cognitive impairment,tardiness,body anxiety and sleep disturbances of the patients with depressive syndromes were higher than that of non-depression patients(P≤0.001).No difference was noted with body weight loss.Conclusion PD patients have a high susceptibility to depression,and duration of PD and H&Y stage appear to be the strongest predictor of depression.

Factors Influencing Outcomes of Methadone Maintenance Treatment

Luo Wei;Wu Zunyou;Wei Xiaoli;Jia Wei;Zhang Quncai;Li Liangliang;Yan Hong

2007, 28(3):  373-377. 

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Objective Methadone maintenance treatment has been considered to be the best practice in the treatment for opiate dependence.It has been agreed by the researchers that treatment retention,employment status,and reduced criminal activity are the essential outcome variables for the treatment.The objective of the study was to recognize factors influencing outcomes of methadone maintenance treatment when patients initially attending the treatment program.Methods Participants were chosen from patients in five methadone maintenance treatment clinics in Xi'an City,Shanxi Province.Patients who had been in the clinic for less than one month were interviewed.There were two interviews in the study.The first one happened when the study began.Patients were asked about the addiction status one month before being recruited into the treatment.The second interview was done in the same population at a time when the participants had been in the treatment clinic for three months.Addiction status in recent one month was investigated in the second interview.Two hundred and eighty-two patients were followed for the two interviews.The Chinese version of Addiction Severity Index was used to evaluate the addiction status of the participants.The Chinese version of Addiction Severity Index was developed on base of the fifth version of the original one and has been proved to be valid and reliable.Unitary linear regression and single variable logistic regression was used when doing simple analysis and multiple linear regression model and multiple logistic regression model were used when doing the comprehensive analysis.Dependent variables included Length of stay in the treatment(actual days of receiving methadone during the first three months in the treatment);decline of times of illegal activities(comparison of times of illegal activities between one month before treatment and three months after treatment);whether having a job(employment status after three months' treatment).Independent variables were seven composite scores of the Chinese version of Addiction Severity Index,that were scores in seven area-medical status,employment status,alcohol abuse,drug abuse,legal status,family/social relationship and psychiatric status;demographic characteristics of the interviewees(age,gender and education level).Results Similar results were found in the unitary regression model and the multiple regression model.Most of the ten independent variables couldn't be included into the model.Age and composite score(psychiatric area) were included in the multiple linear regression with the significant level of 0.10. The Standardized coefficients were0.179(P=0.002) and0.111(P=0.059),respectively.Only gender entered the multiple logistic regression with the dependentvariable of 'decline of times of illegal activities'.The odd ratio was 0.131(95%Confidence Level was 0.024～0.710).Composite score(employment area) and composite score(legal area) were of statistical significance in the multiple logistic regression model with the dependent variable 'whether having a job'.The odd ratio was 0.035(95%Confidence Level was 0.009～0.139)and 0.018(95%Confidence Level was 0.065～0.517),respectively.Patients with young age or severe problems existed in any of the three area: psychiatric area,employment area and legal area were related with negative treatment outcomes.Conclusion Age,composite score(psychiatric area),composite score(employment area) and composite score(legal area) were the factors influencing outcomes of methadone maintenance treatment.Staffs in the clinics should recognize the patients with these characteristics in their early time of coming to the clinic and make proper treatment plan and additional intervention to increase the outcome of the treatment.

The Role of Hyperleptinemia on Atherosclerosis in Patients with Maintenance Hemodialysis

Wang Yinna;Liu Huilan;Yao Ying;Liu Qun

2007, 28(3):  378-381. 

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Objective To investigate the correlation between serum leptin and atherosclerosis and arterial vasodilation in maintenance hemodialysis patients.Methods Fifty maintenance hemodialysis patients and thirty healthy volunteers were enrolled in the study.The levels of serum leptin were determined with enzyme linked immunosorbent assay.Intima-media thickness(IMT) and prevalence atherosclerotic plaques of the extracranial common carotid artery were measured with high-resolution B-mode ultrasonography.The changes in brachial artery diameter during reactive hyperemia were analyzed with ultrasonic examination.Results Both men and women serum leptin levels in hemodialysis patients were significantly higher than those in healthy controls(men(6.13±2.41)μg/L vs(3.25±1.98)μg/L;women(8.98±3.12)μg/L vs(5.79±2.01)μg/L,(P<0.01)).Compared with those in controls,the mean IMT value,percentage of thickened IMT and prevalence of atherosclerotic plaques in hemodialysis patients were significantly increased,while the changes in brachial artery diameter during reactive hyperemia were significantly decreased(P<0.01).Multiple stepwise regression analysis showed that serum leptin was an independent risk factor associated with IMT and changes in brachial artery diameter during reactive hyperemia.Conclusion The level of serum leptin in patients with maintenance hemodialysis is associated with the development of atherosclerosis and dysfunction of arterial vasodilation.Hyperleptinemia may be an independent risk factor for atherosclerosis in patients with maintenance hemodialysis.

Self-control Study of Oxcarbazepine for the Treatment of Childhood Refractory Epilepsy

Yu Yazhen;Tian Zhaoxia;Gao Baoqin;Yang Weili

2007, 28(3):  382-384. 

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Objective To evaluate the long-term efficacy,tolerability and safety of oxcarbazepine(OXC) in children with refractory epilepsy.Methods Thirty-one patients with refractory epilepsy were enrolled in an open add-on study with oxcarbazepine(OXC) to evaluate its long-term efficacy,tolerability and safety.Criteria of enrolment: ① Diagnosed refractory epilepsy by clinic and electroencephalogram in the pediatrics epilepsy outpatient service of Tiantan Hospital.② Aged 3～15 years.③ Criteria for refractory epilepsy: the blood concentration of anti-epileptic drugs in the normal range for at least 2 years by taking the first grade anti-epileptic drugs regularly for at least 2 years,but failed to control seizure and improve the everyday life,with episodic attacks of over 4 times per month.Excluding criteria: ① Tumor and systematically deteriorating diseases of central nervous system.② Accompanying heart,liver,kidney,thyroid gland or blood diseases.③ Carbamazepine allergy.④ Filed to be followed up regularly.The treatment course was 12 months.The investigators should know about the currentry taking anti-epileptic drugs,frequencies and types of attacks three month before starting the investigation.The items examined included nerve image,electroencephalogram and laboratory tests.OXC was added to the current anti-epileptic drugs.The initial dose was 4～5 mg/(kg·d^-1).The dosage was increased by one time dose every 2 weeks,and the increment should not exceed 10 mg/(kg·d^-1).The maintenance dose was(25～45) mg/(kg·d^-1).The seizure frequencies,adverse events three months before and twelve months after therapy were compared with the seizure frequencies of the in 12 weeks taken as the base line,and the observed results were rank remarkably efficient(the frequency decrement ≥75%);efficiency(the frequency decrement ≥50%);not efficiency: the frequency decrement <50%;and deteriorate(the frequency increased by≥25%).The blood and urine routines,the heart,liver and kidney functions,serum lipid and sodium levels,blood concentrations of anti-epileptic drugs,and electroencephalograms before and after therapy were compared also.Results Of the 31 patients,there were 13 partial seizure cases,7 generalized seizure cases,9 symptomatic generalized seizure cases and 2 lennox-Gastaut syndrome cases.At 6 months of treatment,oxcarbazepine(OXC) can significantly reduce seizure frequency compared with the pre-OXC period.It seemed that oxcarbazepine(OXC) was most efficacious in partial seizure.It showed that the seizure frequencies before and after therapy were significantly different(P<0.05).However,there wa significant difference at 6 and 12 months after therapy(P>0.05).Five patients(5/31,16%) became seizure-free,three patients(3/31,9.7%) remarkably efficient,six patients(6/31,19.4%) were efficient.The total effective rates(seizure frequency decreased by ≥50% were 14/31(45.2%).In addition,the numbers of interictal epilepsy discharge in EEG of 16 patients were decreased after OXC administration compared with with pre-OXC period.Two patients had dropped out because of economic problem and/or adverse events.Adverse reactions were found in 12.9%(4/31) of patients,the common adverse events were dizziness,headache,sommolence,nausea,fatigue and rashes.Adverse reactions were generally mild,and disappeared when dose was increased slowly.Conclusion The efficacy of OXC is sustained with good safety and tolerability profiles during the treatment of 12 months in children with refractory epilepsy.Safety of OXC in younger children(≤50 years old) need to be further validated.

The Sensitivity Analysis of Single Fiber Electromyography in Ocular Myasthenia Gravis

Liu Ying;Gao Yuhong;Jing Yun;Yu Fengchun;Lu Yang;Lu Changfeng;Zhang Xiaojun

2007, 28(3):  385-387. 

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Objective To study and estimate the value of SFEMG of the frontalis muscle in the diagnosis of ocular myasthenia gravis(OMG),comparing the sensitivity of the SFEMG of the frontalis muscle,repetitive nerve stimulation(RNS),blood concentration of antibody to the acetylcholine receptor(AchR-Ab) and neostigmine test.Methods In insulated room of 25 ℃,we performed SFEMG of the frontalis muscle in 27 patients with OMG who attended Beijing Tongren Hospital from August 2006 to January 2007 and 12 controls.RNS,AchR-Ab titration and neostigmine test were also performed.Results 1) Eighteen of twenty seven patients had abnormal recordings in SFEMG of the frontalis muscle,the mean jitter ranged from 20 to 97 us(average 45.4±21.0 μs),the ratio of jitter >55 μs was(24.2±24.5)% and percentage of blocks was(1.9±4.4)%,in the controls,the mean jitter ranged from 19 to 40 μs average(27.4±6.6 μs);9 of 27 patients were abnormal in low frequencies of RNS,among which 7 patients had abnormality in facial nerve,one in facial and axillary nerve and one in axillary nerve;AchR-Ab titersns were increased in 8 patients;23 of 27 patients were positive in neostigmine test.2) Sensitivities of ACh-Ab titration(29.6%) in patients with OMG were remarkably lower than SFEMG of the frontalis muscle,sensitivity of RNS(33.3%),neostigmine test(85.2%) was not different from SFEMG of the frontalis muscle.Conclusion SFEMG of the frontalis muscle shows higher sensitivity for the diagnosis of OMG than RNS and AchR-Ab titration.

Study on the Minimal Effective Dose of Bupivacaine in Spinal Anesthesia Caesarean Section

Zhang Shuo;Zhang Wenyu;Zhang Guogang;Xu Mingjun

2007, 28(3):  388-391. 

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Objective To study the minimal dose and safty of bupivacaine spinal anesthesia in caesarean section.Methods Ninety patients ready for caesarean section were recruited and randomly divided into three groups(groups A,B and C with 30 cases for each group) and received combined spinal-epidural anesthesia(CSEA).Patients were injected intrathecally with 0.5% iso-proportional bupivacaine 5 mg(group A),6 mg(group B) and 7 mg(group C).Additional 2% lidocaine were added if necessary.If the systolic pressure was less than 90 mmHg or the diastolic pressure was 20% less than standard values,10 mg ephedrine would be used to correct it.After these steps,all medical indexes were abserved as follows: I-D time(interval skin incision to delivery),U-D time(interval from uterine incision to delivery) and anesthesia & operation time were recorded;meanwhile the level of anesthesia,bromage scores,blockade effectiveness,and the effectivness of additional epidural lidocaine were recorded.As well as patients'heart rate,blood pressure,and blood oxygen saturation at each postanesthetic time interval.Apgar scores of the neonate,side effects such as panting,nausea,vomiting and traction reflex of the gravide were also observed.Results Statistical analyses were performed by SPSS 10.0.ANOVA(analysis of variance) and Chi-square tests were used.The standard level of significance used to justify a claim of a statistically significant effect was 0.05.Major results were listed as follows.Anesthesia and operation times of groups B and C were(43.30±11.50)min and(43.50±10.90)min respectively, which were shorter than group A(51.20±12.8)min(P<0.05),but there was no difference about I-D timeand U-D time in three groups(P<0.05).Levels of anesthesia in groups A,B,C were T7.75±1.71、T6.27±1.85 and T6.21±1.42,respectively.Group B and group C were higher than group A(P<0.01).Bromage scores of groups A,B,C were(2.07±0.77)、(2.41±0.63) and(2.81±0.49),respectively.Group C were superior to group A(P<0.05).Blockade effect of group B(1.24±0.44) and group C(1.15±0.37) were superior to group A(2.11±0.69)(P<0.01);additional use of epidural 2% lidocaine occured more often in group A(67.9%) than in group B(24.1%)and group C(11.5%)(P<0.01).Systolic and diastolic blood pressures at postanesthetic 5th minute in group C(100.2±24.0 mmHg,(62.3±16.6)mmHg correspondingly were lower than group A(110.2±12.2)mmHg,(70.7±10.3)mmHg respectively(P<0.05),but there was no difference at other times among three groups(P>0.05).Also there were no difference in terms of heart rate,SPO₂ and Apgar scores of the neonate among three groups(P>0.05).Nausea and panting in group C(38.5%,57.7%) were more often than in group A(14.3%、7.1%) and group B(13.8%、6.9%)(P<0.05,P<0.01 respectively).Conclusion The dosage of bupivacaine is substantially related to its side effects and toxicity.0.5% bupivacaine 6 mg is the minimal and reliable effective-dose to be used in caesarean section with lumbar anesthesia,and is highly recommended.

Gamma Knife Radiosurgery for Nonfunctioning Pituitary Adenoma Involving Visual Pathway

Wang Meihua;Liu Ali;Liu Peng;Luo Bin;Sun Shibin;Pan Jian

2007, 28(3):  392-394. 

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Objective A retrospective analysis assessed the efficiency of gamma knife radiosurgery for nonfunctioning pituitary adenoma involving visual pathway.Methods Imaging and endocrinological evaluations following gamma knife radiosurgery were performed in 295 patients with nonfunctioning pituitary adenoma,and the patients were divided into two groups according to the tumor involving visual pathway or not,followed up from 6 to 106 months(average 35 months).The mean margin dose of 82 cases of visual pathway involved patients was 13.4 Gy and the visual pathway suffered an average radiation dose of 8.6 Gy(7 to 10 Gy).To the 213 cases of visual pathway non-involved patients,the average margin dose was 14.7 Gy and the visual pathway suffered 6.2 Gy(3 to 9 Gy).Results 47 cases(57.3%) of tumor volume decreasing,30 cases(36.6%) of remaining unchanged,and 5 cases(6.1%) of increasing were found in the group of 82 visual pathway involved patients,2 cases presenting visual impairment.149 cases(70.0%) of tumor volume decreasing,61 cases(28.6%) of remaining unchanged,and 3 cases(1.4%) were found in the group of 213 visual pathway non-involved.The overall tumor control rate was 97.3%.7 patients suffered new hormone deficits following gamma surgery.Conclusion Gamma knife radiosurgery is efficient and safe for nonfunctioning pituitary adenomas,especially for visual pathway non-involved patients with few complications.But serious consideration should be taken for patients with obvious visual pathway compression.An intensive dose plan is essential to reduce the incidence of optic neuropathy.

The Effects of Contrast Medium on the Pancreatic Microcirculation in Rats with Severe Acute Pancreatitis

Cao Zhanyu;Sun Jiabang

2007, 28(3):  395-398. 

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Objective As a good diagnostic means,contrast-enhanced computed tomography(CECT) has been used for evaluating acute pancreatitis,but,it is uncertain whether the contrast medium(CM),which must be used in CECT,has adverse effects to the microcirculation of pancreas with severe acute pancreatitis(SAP).The purpose of this study is to observe the effects of contrast media on microcirculation of pancreas with SAP by measuring volume of blood flow and evaluating changes in pathology of pancreas in rat SAP models,which has been intravenously injected CM.Methods SAP models of wistar rats were divided into 5 groups randomly: control group,SAP group,SAP with normal saline injecting group,SAP with 76% Cardiografin injecting group and SAP with ultravist-300 injecting group. Then,volume of blood flow into the pancreas was measured and changes in pathology were evaluated to show the influence of CM on microcirculation of pancreas with SAP.Results Comparing with SAP group,the other 3 groups had no significant differences on volume of blood flow and pathologic changes(P<0.05), and there was no significant difference among the 4 study groups(P>0.05).Conclusion This study shows that CM has no adverse effects on the microcirculation of rat pancreas with SAP and would not worsen the disease.

The Clinical Analysis on Efficacy of High Ligation Combined with Endovenous Laser Treatment in Greater Saph

Jia Yulong;Li Na;Wang Yan;Cheng Shi;Song Maomin

2007, 28(3):  399-401. 

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Objective To investigate the efficacy of high ligation combined with endovenous laser treatment(EVLT) in greater saphenous vein varicosity.Methods The clinical data of 218 cases of greater saphenous vein varicosity who underwent high ligation combined with EVLT during July 2003 to July 2006 were retrospectively reviewed,and were compared with the 100 cases as control who were operated on with the common surgery during 1998 to 2002.Results All of the patients who underwent the EVLT had healing by first intention without infection.Among most of them the symptoms of lower extremity disappeared or were alleviated,no motion disturbance during one month postoperative followup.There were no deep venous thrombosis and deep vein injury among them.Conclusion High ligation combined with EVLT is advantageous in terms of less incision,minimal hemorrhage,shorter operation time,more rapid recovery and shorter hospitalization time compared with common operation.Further more the complication is decreased and the recurrence is lower.However it is still imperfect.

Clinical Value of Lumbar MR Myelography for Degenerative Lumbar Spine Disorders

Zhang Lehui;Zhao Yi;Shao Feng

2007, 28(3):  402-405. 

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Objective To assess the practicability and reliability of lumbar MRM and analyze the clinical value of lumbar MRM for degenerative lumbar spine disorders.Methods One lumbar intervertebral disc(L₃₄,L₄₅,L₅S₁) as well as two nether lumbar nerve roots make up an investigative unit. One hundred and eight investigative units of 36 patients with degenerative lumbar spine disorders were examined with computed tomography(CT) and magnetic resonance imaging(MRI) and MRM.All patients were treated surgically.The evaluation of the practicability included the quality of image,security and cost.On the basis of clinical diagnosis,we compared the sensitivity and specificity of CT,MRI and MRM by χ² test in order to assess the facticity of lumbar MRM.Results Thirty-four(94.4%) of the patients' images were excellent.All were examined without injury or side-effect.The average examination time is eighteen minutes and the cost is 1 080 RMB.Statistical results showed that MRM as a whole appears to be as sensitive as MRI or CT but more specific than MRI or CT in diagnosing degenerative lumbar spine disorders.Conclusion MR myelography has many technical characters:(1) Technical improvements make the operation simple and more convenient.(2) The quality of images is excellent.(3) The security is good.(4) The integrative cost is no higher than other means.(5) MRM not only has an obvious advantage in locating the lesion of degenerative lumbar spine disorders,but also helps to distinguish the kind of degeneration.(6) The specificity is higher and so is the sensitivity.Therefore,MRM has important clinical value in the diagnosis and operative planning of degenerative lumbar spine disorders,but it has not been regarded as a conventional examination now.

PCEA Contributes to Prevention and Treatment Post-epidural Puncture Headache

Che Xiangming;Xu Mingjun

2007, 28(3):  406-408. 

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Objective To evaluate the feasibility of injecting saline continuously by PCEA for preventing and treating post-epidural puncture headache(PEPH).Methods Forty-six patients who had had their dura punctured through,were divided into 2 groups;Group A(n=37): in this group patients received combined spinal-epidural anesthesia in another interspinal space if spinal arachnoid was punctured through,and saline injected continuously into the epidural space post-operatively;Group B(n=9): no injection into epidural space,patients received general anesthesia after spinal arachnoid was punctured through.The incidence of post-epidural puncture headache were recorded in all patients.Results The incidence of PEPH in group A was 16.2%,The incidence of PEPH in group B was 100%.Conclusion Injecting saline continuously by PCEA is effective in preventing and treating post-epidural puncture headache.

Accurate Placement of Central Venous Catheters in Sitting Position Neurosurgical Operation

Ma Ting;Cai Bing;Lan fei;Wei Limin;Wang Kejie

2007, 28(3):  409-411. 

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Objective To place the central venous catheters accurately guided by the intracardic electrocardiography(ECG) and investigate the effects of aspirating air embolus in sitting position neurosurgical operation.Methods Thirteen sitting operation patients who received a central venous catheter to aspirate air embolus were enrolled.The central venous catheter was accurately placed by using guiding wire as internal electrode to record intracardiac signals.The position of the catheter was assessed through the changes in intracardiac ECG and re-checked with X-rays after operation.When air embolus was detected,it was aspirated through the central venous catheter and the volume of air calculated.Results The technique of using intracardiac ECG to guide the placement of central venous catheter was accurate.Seven air emboli were detected in 13 patients.The success rate of aspiration was 100% and the volumes of aspirated air were from 0.5～20 mL.Conclusion The technique of using intracardiac ECG guidance to detect central venous catheter tip is convenient and accurate;Air embolus can be aspirated effectively.

The Variety of Vascular Endothelial Growth Factor Expression in Endometrium of Tamoxifen User

Yang Baojun;Xing Fengling;Feng Limin;Zhang Hua;Sun Haimei

2007, 28(3):  412-414. 

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Objective To investigate the bimolecular effects of tamoxifen on the uterine endometrium in women with breast cancer.Methods Using immunohistochemistry(LSAB) on formalin-fixed paraffin-embedded endometrial sections to study the variety of vascular endothelial growth factor(VEGF) in endometrium of postmenopausal patients with breast carcinoma who took tamoxifen more than two years and in endometral adenocarcinoma,with prolapse uterine endometrium as normal controls.Results The VEGF staining indices were significantly higher in endometrium of tamoxifen user and endometrial adenocarcinoma than in controls(P<0.05).The VEGF in the endometrium tamoxifen user was markedly increased, as much as that of endometrial adenocarcinoma.Conclusion The High-score of VEGF expression of the endometrium in women who have been treated with tamoxifen for breast cancer may be correlated with tamoxifen,which as the mechanism of oestrogen and anti-oestrogen action has the ability to affect the endometrium angiogenesis.Tamoxifen is able to induce pathological changes in endometrium.

Diagnosis and Treatment of Malignant Tumor of Lacrimal Sac Tumor

Zhang Xuxiang;Qu Dongyi;Qu Qiuyi

2007, 28(3):  415-417. 

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Objective To report a series of 5 primary lacrimal sac tumors and present an algorithm in managing this rare condition.Methods A retrospective review of the clinical,radiological,and pathological records of 5 patients with primary lacrimal sac tumors was conducted.Results Of the five patients,all had epiphora or mass above the medial canthal tendon.Four had bloody epiphora.Three cases of computerized tomography,one case of dacryocystography,B ultrasonic as well magnetic resonance.There was extension into the anterior ethomoid air cells in only one case.The commonest tumor was squamous cell carcinoma(two cases),followed by one case of transitional cell carcinoma,basal cell carcinoma,undifferentiated carcinoma.Treatment modalities included surgery,radiotherapy,one patient had a relapse,En bloc resection was performed using a lateral rhinotomy,medial maxillectomy.but four patients remained without evidence of recurrent tumor.Conclusion Bloody epiphora,chronic dacryocystitis that irrigates freely and mass above the medial canthal tendon are main signs of malignant lacrimal sac tumor.Early diagnosis with malignant lacrimal sac tumor is possible combining with adjunctive examination and biopsy specimen.It can be distinguished with ethomoid carcinoma combining with clinical performance and dacryocystography,computerized tomography as well magnetic resonance.The treatment of malignant lacrimal sac tumors is en bloc resection and postoperative irradiation.

Treatment of Adenoidal Hypertrophy by Nd:YAG Laser Under Fiberoptic Laryngoscope

Feng Lei;Li Xia;Sun Rushan;Jiang Pei

2007, 28(3):  418-419. 

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Objective To study the clinical efficacy of Nd:YAG laser in treating adenoidal hypertrophy.Methods By vaporizing or coagulating with Nd:YAG laser under fiberopic laryngoscope,857 adenoidal hypertrophy patients with nasal obstruction and/or auditory tube compression were treated,all were under surface anesthesia.Results 761 cases were clinical cured,78 cases were improved,18 cases were failed.The cure rate was 97.9%.Conclusion The application of Nd:YAG laser in adenoidal hypertrophy is easy to operate,and minimally invasive.It is reliable and effective for most cases.