Table of Content

24 April 2007, Volume 28 Issue 2

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The Clinical Application of DNA Chip for Common Gram-positive Bacteria Identification and Drug Resistance Detection

Wang Yajie;Wang Can;Fang Fang;Shi Congning;Jiang Di;Zhang Guojun;Lü Hong;Yang Huawei;Chang Zheng

2007, 28(2):  140-144. 

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Objective To study the clinical application of DNA chip for common Gram-positive bacteria identification and drug resistance detection. Methods 445 Gram-positive strains were identified with physiological and biochemical methods, and were detected of drug resistance with diffusion disk method for susceptibility test. Then the strains were blindly detected with using DNA chip for identification and drug resistance. The data were analyzed using SPSS 10.0 for Windows software. Results The sensitivity of all identification indexes of DNA chip were higher than 96%, and all of their specificity were higher than 98%. The sensitivity of all drug resistance indexes of DNA chip were higher than 94%, and all of their specificity were higher than 90%. The chi square test values of all indexes were higher than 0.05, showing that there was no significant difference between DNA chip and the clinical routine methods. The Kappa values of all indexes were higher than 0.75, indicating that the coincidence between the two kinds of methods was good. Conclusion There is clinical potential of gene chip technology for bacteria identification and drug resistance detection.

The Effects of HDTIC Compounes Isolated from Astragali Radix on the Expression of p16 and p21

Wang Peichang;Zhang Jian;Zhang Zongyu;Tong Tanjun

2007, 28(2):  145-149. 

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Objective To investigate the effects of HDTIC-1 and HDTIC-2 isolated from Astragali Radix on the expression of p16 and p21 of human fetal lung diploid fibroblasts(2BS) so as to explore the mechanism of HDTIC delaying replicative senescence. Methods To analyse the expression levels of p16 and p21 of 2BS cells cultured with DMEM containing HDTIC by RT-PCR and western blot. Results 1)There was no expression of p16 in PD 45 2BS cells incubated with HDTIC.In PD 56 2BS cells cultured with HDTIC, there was a weak mRNA expression of p16, whereas, the protein expression of p16 was not observed.However,an obvious expression of p16 was seen in the control senescent cells. 2)For the cells cultured with HDTIC,there was a weak expression of p21 in the middle age and a strong expression of p21 in the senescent cells.Additionally,no difference of p21 level between the HDTIC-cultured cells and the control cells was seen. Conclusion There was no effect of HDTIC compounds on the expression of p21.But p16 expression of 2BS cells was strongly inhibited by HDTIC,which may contribute to their delaying replicative senescence.

The Comparison of Automatic Microorganism Analyzer and Gene Analysis in Identification of Coagulase Negative Staphylococcus

Wang Mei;Wu Wei;Lu Xinxin

2007, 28(2):  150-153. 

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Objective To compare the sensitivity and specificity in identifying coagulase negative staphylococcus(CNS) between Vitek32 microbic system and 16S rRNA sequence analysis. Methods 70 strains of CNS isolated from Intensive Care Unit(ICU) the time period from during December 2004 through November 2006 were chosen to perform convention bacilliculture, then they were classified as Vitek32 microbic system and 16S rRNA sequence analysis, and the superiority and shortage of these two identification methods were compared. Results The identifying results were divided into three groups according to Vitek32 system standard. Group Ⅰ,'unacceptable group' %id<80, had 25 strains; Group Ⅱ,'acceptable group'%id 80～90, had 35 strains; and Group Ⅲ,'Good group'%id 90～98, had 10 strains. The standard strain was the Staphylococcus aureus(ATCC29213). All these strains were carried out 16S rRNA gene sequencing respectively. The coincidence rates of group Ⅰ, group Ⅱ and group Ⅲ using these two methods were 64%, 74.3% and 100%, respectively. The overall coincidence rate was 74.4%. Conclusion 16S rRNA sequence analysis could make up the limitation of Vitek32 system in identifying CNS. It will play an important role in identifying the microorganisms that are difficult to culture or those could not be cultured.

Anti-Nuclear Antibodies and Its Clinical Implication in Patients with Hepatitis B Virus Infection

Zhang Haiping;Yan Huiping;Feng Xia;Tan Yufen;Liu Yan;Li Weihua;Ma Dongmei;Zhao Yan;Cui Dan

2007, 28(2):  154-157. 

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Objective To investigate the characteristics of anti-nuclear antibodies and its clinical implication in patients with hepatitis B virus infection. Methods 569 patients with hepatitis B virus infection including 214 with chronic hepatitis B, 283 with liver cirrhosis and 72 with hepatocellular carcinoma were recruited. 30 healthy volunteers were as control. Indirect immune fluorescent assay(IIF) was used to detect the serum anti-nuclear antibody(ANA), anti-mitochondrial antibody(AMA), anti-smooth muscle antibody(SMA) and anti-liver special protein(LSP). Western blot was used to detect ANA profile. Results 1) 245 of the 569 patients with Hepatitis B virus infection(43%) were ANA positive, 90/214(42.1%) in CHB group, 108/283(38.2%) in LC group, 47/72(65.3%) in HCC group. HCC had a significantly higher percentage of ANA than CHB, LC and healthy volunteers(6.7%). 2) Among the 3 groups, low titer (1:100) of anti-nuclear antibodies was more common. Although a rage of fluorescent patterns was observed, the percentage of granular fluorescence was highest in the 3 groups. The percentage of anti-nucleolar antibody in HCC group(19.1%) was higher than CHB(4.4%) and LC(18.5%) (P<0.01). 3) ANA profile was detected in 45 patients with titer of ANA ≥1:320. dsDNA was positive in 2 patients with LC , and SSA in 1 patients with LC. 4)In terms of the serum levels of ALT, AST and virus load there were no significant differences between titer of ANA 1:100 and 1:320 in each group. Conclusion Anti-nuclear antibodies(ANA) were main auto-antibody in patients with hepatitis B virus infection, especially in patients with HCC. The titer of ANA had no significant implication with liver damage.

Serum Levels of VEGF,VEGF C and uPAR in Patients withOral Squamous Cell Carcinoma

Yu Xuemei;Liu Xiaoyong;Wang Yajie;Ding Li

2007, 28(2):  158-161. 

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Objective The serum levels of VEGF, VEGF C and uPAR in patients with oral squamous cell carcinoma(OSCC) were determined to study the relationship between VEGF, VEGF C and uPAR serum levels and pathological parameters or lymphatic metastasis in OSCC. Methods Serum levels of VEGF, VEGF C and uPAR were measured with quantitative enzyme-linked immunosorbent assay in 91 patients with OSCC and in 50 healthy individuals as control group. Results Significantly elevated levels of VEGF, VEGF C and uPAR were observed in OSCC compared with control group. No statistically significant correlation could be found between the serum levels and histological grading, infiltrative mode or nodal status in OSCC. Conclusion Serum determinations of VEGF, VEGF C and uPAR may provide laboratory data for monitoring the occurrence and development of OSCC, but have no significance in predicting lymphatic metastasis.

Clinical Investigation of Serum Glial Fibrillary Acid Protein in Patients with Acute Cerebral Infarction

Tan Yanguo;Xue Yinghong;Xu Bin;Li Weidong

2007, 28(2):  162-165. 

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Objective In patients with acute cerebral infarction serum glial fibrillary acidic protein(GFAP) level and its temple profile was determined to investigate the relationship between GFAP level and the size of brain lesions, neurological status(NIHSS) and functional outcome(Barthel Index, BI). Methods Self-established biotin-avidin multi-layers magnifying solid phase enzyme immunoassay was used to determine serum GFAP level in patients with acute cerebral infarction at the time of 48 h, the fifth day, the seventh day and the fourtheen day after onset of stroke. All patients'neurological status was evaluated by the National Institutes of Health Stroke Scale (NIHSS) at corresponding time points, and functional outcome was evaluated by BI at discharge from the hospital. Results For all patients observed, serum GFAP level increased markedly compared to that of control group at the time of 48 h after onset of stroke(P=0.015). With time elapsing, it decreased, and restored to normal level at the seventh day(P=0.056), but increased again at the fourteen day compared to that of control group(P=0.027) . There was significant difference for serum GFAP levels among patients with larger, middle, smaller size of brain lesion and normal control group at 48 h and the fifth day respectively after onset of stroke(P=0.032, P=0.048). Patients with larger size of brain lesion have relatively higher level of serum GFAP. There was significant linear negative correlation between serum GFAP level and NIHSS score at the fifth day or the seventh day, respectively(P<0.05),and there was also significant linear positive correlation between GFAP on the fifth day and the seventh day and BI score at discharge(P<0.05). Conclusion GFAP level in blood of patients with acute cerebral infarction increase markedly, which might be a useful marker for disease assessment and prognosis predicting.

Study of Naturally Occurring Mutations in Open Reading Frame of CCR5 in Hong Kong Chinese

Zhao Xiuying;Li Ruishan;Huang Jiaqing;Chen Zhiwei;Huang Chun;Zheng Bojian

2007, 28(2):  166-169. 

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Objective To determine the polymorphism of HIV co-receptor, CC chemokine receptor 5(CCR5) in Chinese and its effects on HIV infection. Methods Two groups of Chinese residing in Hong Kong were recruited: ① 785 HIV-healthy donors and ② 314 HIV+ patients. CCR5 mutations were identified from the study subjects by full sequencing of the open reading frame(ORF). Results Ten mutants of CCR5 ORF were identified, seven of which were non-synonymous. Two previously reported mutants, R223Q and 299(FS) were proved to be the most prevalent. These alleles did not show significant difference between HIV patients and healthy controls. However, the novel mutant G106R might affect the conformation in the second extra-cellular loop(ECL2) of CCR5 and change its co-receptor function. Conclusion Mutation in the CCR5 ORF is quite prevalent in ethnic Chinese. However, the identified mutants do not affect HIV infection from epidemiological level. Structural analysis had indicated that the novel mutant G106R in the third trans-membrane domain might affect its function as co-receptor for HIV.

The Clinical Value of Adenosine Deaminase Level for the Diagnosis of Some Neurologic Diseases

Chen Yan;Huang Zeyu;Wang Suping;Wang Yajie;Kang Xixiong

2007, 28(2):  170-172. 

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Objective To study the clinical value of Adenosine deaminase in cerebrospinal fluid and serum for the diagnosis of some neurogical diseases. Methods With Peroxidase assay to detect the Adenosine deaminase level in serum and cerebrospinal fluid in 201 cases of the neurological diseases(benign tumors, malignant cancer, multiple sclerosis, cerebral infarction, hydrocephaly, encephalic angiona, brain bemorrhage, spinal cord tumor, virus meningitis) as well as 24 normal persons. With t test or rank test to statistical analysis for the cases with neurological diseases which were divided into 9 groups and normal group. The cases of all neurological diseases groups were compared with each other. Results There was no significance for all the cases with adenosine deaminase level in serum from the neural disease. The adenosine deaminase levels in CSF from neurological diseases exception block in brain were significantly higher than those in normal group(P<0.01). There were statistic significances. Adenosine deaminase levels in CSF from malignant cancer group were higher than those in benign tumors group(P=0.037). Adenosine deaminase levels in CSF from multiple sclerosis were of statistic significances than those in spinal cord tumor, meningitis and cerebral hemorrhage(P<0.05). Conclusion Adenosine deaminase level in serum is of no significance in the diagnosis and identification. The determination of adenosine deaminase in cerebrospinal fluid is helpful in the diagnosis differentiation of some neurological diseases, but it is of no special significance in diagnosis.

Resistant Genes Detection and Homologous Analysis ofVancomycin Resistant Enterococcus

Liu Zhiyuan;Xu Shuzhen;Yu Yanhua

2007, 28(2):  173-175. 

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Objective To detect vancomycin-resistant genes of enterococcus, and to analyses antibiotic resistant phenotype and genotype. Methods Susceptibility of vancomycin and other antibiotics were performed with agar diffusion methods. Vancomycin resistant genes and homologous analysis were detected with PCR and RAPD methods, respectively. Results 6 strains of Enterococcus gallinarum and 4 of Enterococcus faecium were identified in the study, all of which carried vanA gene. According to RAPD results, 10 strains were divided into 7 genetypes. Conclusion Vancomycin resistance in enterococcus is mainly mediated by vanA. RAPD is the effective methods to analyze the homogeneity of vancomycin resistant enterococcus.

CYP2C19/CYP2D6 Genetic Polymorphisms in Chinese Healthy and Schizophrenia Populations

Zhou Jian;Kang Xixiong;Lü Hong;Wang Yajie;Cheng Xueyan;Wang Gang

2007, 28(2):  176-179. 

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Objective To investigate whether cytochrome P450 2C19(CYP2C19) /P450 2D6(CYP2D6) genetic polymorphisms were associated with schizophrenia in Chinese subjects. Methods 88 schizophrenics meeting criteria ICO-10 were enrolled in the study group and 283 healthy volunteers were involved in the control group. Polymerase chain reaction(PCR) and DNA sequencing were used to examine the CYP2C19(CYP2C192/CYP2C193 ). Use the same methods to detect CYP2D6(C100T) genetic polymorphisms of 87 schizophrenics meeting criteria ICO-10 and 93 healthy volunteers. The patients and healthy volunteers were divided into three groups accoding to the CYP2C192(G681A) genotypes(G/G, G/A, A/A), the same as CYP2C193(G636A) and CYP2D6(C100T). Results Six different allelic variants of CYP2C19 gene were observed in the control group and seven different allelic variants of CYP2C19 gene were detected in the schizophrenia group. There is no statistically significant difference in the allele frequencies of CYP2C192(χ²=4.954, P>0.05)and CYP2C193(χ²=2.334, P>0.05) between schizophrenia and healthy volunteers. We found the frequency of the homozygote of CYP2C192 in schizophrenia was conspicuously higher than that of healthy volunteers, There is no significant difference between two groups in CYP2D6(C100T) genotypes and alleles. Conclusion Although there is no evidence to show the correlation between CYP2C19/CYP2D6(C100T) genetic polymorphisms and etiology of schizophrenia, the homozygote of CYP2C192 is possible a genetic susceptibility factor in Chinese subjects with schizophrenia.

New Development of Clinical Laboratory Medicine in USA

Xu Min

2007, 28(2):  182-184. 

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In the past twenty years, with the development of technology and wide application of computer technology, automation has become reality in biomedical fields. Currently the challenge in laboratory medicine is to develop new testing methods with higher sensitivity and efficiency. This review introduced several new technologies including Luminex^RxMAP^TM, Pyrosequencing, and Nanoparticle technology. These technologies have been entering the clinical laboratory in the past few years. In addition, the quality control, standardization, and organization of future clinical laboratory are also discussed.

Research Advances of Laboratory Diagnosis on Helicobacter Pylor

Sun Yanyan;Wang Yajie;Kang Xixiong

2007, 28(2):  185-188. 

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Helicobacter pylor can cause chronic gastritis and peptic ulcer. It is one of the risk factors of gastric cancer. There is correlation between Hp infection and other associated diseases. There are many examination methods about Hp infection. With announcement of whole genome sequence of Hp and the development of immunology, gene theory and proteomics, the improvement of Hp examination has become the research focus. This review summarizes and introduces the research progress of Hp examination.

Quality Assessment of Traditional Literature Reviews on TCM Treatment:Diabetic Nephropathy as an Example

Ai Yanke;Wan Xia;Liu Jianping

2007, 28(2):  189-191. 

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Objective To evaluate the quality of traditional literature reviews of traditional Chinese medicines(TCM) for diabetic nephropathy to explore the existing problems and to provide basis for methodological improvement. Methods According to the principles and methods of clinical epidemiology and evidence-based medicine and referring the 18 items of QUAROM Statement, we developed a data extraction form. Review articles of TCM for diabetic nephropathy published between Jan 1989 and June of 2005 in Chinese journals were evaluated. Results In total, 108 reviews were included for evaluation in this paper. 99% of the articles did not specify the objectives, and all articles did not describe the sources of cited references, and no information on study design and validity evaluation. Conclusion Traditional reviews are of wide topic areas and lack of definite objectives, inclusion criteria, and may suffer from selection bias, citation bias and publication bias, and lack of qualitative or quantitative synthesis of data. Therefore, the conclusions are not evidence based.

Salvianolic Acid-B Effects on TGF-β1/ERK Signaling Transduction in NIH/3T3 Fibroblast

Tao Yanyan;Wang Xiaoling;Liu Chenghai

2007, 28(2):  192-195. 

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Objective To investigate the mechanism of salvianolic acid B(SA-B) against liver fibrosis related to TGF-β/ERK signal transduction. Methods NIH/3T3 fibroblasts were divided into 4 groups: normal, model and treated groups with lower and high dosage of SA-B. Except of the normal group, the cells were incubated with 100 pmol/L TGF-β1 for 24 h, and the treated group in-cubated with 1 μmol/L or 10 μmol/L SA-B in 0.5% FBS/M199 at the same time respectively. The NIH/3T3 fibroblast viability was observed with MTT assay. The collagen type I and plasminogen activator inhibitor-1(PAI-1) proteins were analyzed with Western blot. The proteins of TGF-β receptor type Ⅰ and Ⅱ (TβR-Ⅰ, TβR-Ⅱ), ERK1 and its phosphorylation(P44 and P42) were analyzed with Western blot. Results TGF-β1 increased the expression of collagen type Ⅰ protein, improved its PAI-1 production, both of TβR-Ⅰ and Ⅱ protein expression and ERK phosphorylation. MTT assay showed SA-B had no effect on the viability of NIH/3T3 fibroblasts, While SA-B inhibited TGF-β1 stimulated collagen and PAI-1 production in dose-dependent manner, decreased TβR-Ⅰ protein expression, and inhibited ERK phosphorylation, and 10 μmol/L SA-B had a much better effects than 1 μmol/L SA-B. Conclusion SA-B antagonizes the profibrogenic effects of TGF-β1 by inhibiting TβR-Ⅰ expression and ERK phosphorylation, and these actions are associated with the mechanism of SA-B effect against liver fibrosis.

Effects of Fuzhengjiangan Formula on Collagen Metabolism in Rats withImmune Hepatic Fibrosis

Huang Xiangan;Chen Yanli;Sun Lihong;Song Chongshun;Cui Ning;Xu Hongyan;Ren Ying

2007, 28(2):  196-199. 

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Objective To investigate the effects of Fuzhengjiangan Formula(FZJGF) on collagen metabolism in liver fibrosis rat model induced by the human serum albumin. Methods The rat models with immune liver fibrosis were induced by the human serum albumin. Rats were treated with saline, FZJGF(crude drugs 9.85,39.4 g/kg, two dosage groups) and colchicine(0.000 1 g/kg). The contents of laminin, hyaluronic acid and collagen type Ⅳ in rat serum were measured with radioimmunoassay method. The level of hydroxyproline in liver tissue was detected with chemistry method. The pathological changes of liver tissue were observed with HE and Von Gieson staining. The contents of collagenous fiber were determined with image quantitive analysis system. Results FZJGF could significantly decrease the contents of LN, HA and Ⅳ-C. At the same time, it depressed the level of Hyp in rats liver. The pathological improvement of pathologyical changes were observed and FZJGF could markedly alleviate deposition of collagenous fiber, reduce liver pseudoluboli. The image quantitive analysis showed that the contents of collagenous fiber of liver significantly decreased after the treatment using FZJGF. Conclusion FZJGF could affect the collagen metabolism in liver to inhibit formation and development of rat hepatic fibrosis induced by the human serum albumin.

Effects of Xiaoshi Fanshi Powder on Hepatic Pathological Damage Caused by Immune Hepatic Injury Model in Mice

Zhang Qiuxia;Zhao Hui;Yu Nan;Yan Xiaoyuan;Xu Meng;Xue Pu

2007, 28(2):  200-202. 

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Objective To study the effects of different xiaoshi Fanshi Powders which consist of different components on hepatic pathological damage caused immune hepatic injury in mice. Methods Firstly, ICL mice were selected and randomized into seven groups: normal control group,model group, Huoxiao Baifan group, Huoxiao Lufan group, Mangxiao Baifan group, MangxiaoLufan group, and bifenbate group. Secondly, one day after intravenous injection of BCG, the equal amount of saline was given for gastric perfusion to the normal control group and model group. The corresponding medicines were given for gastric perfusion to the other groups. At last, The liver was reserved for routine light microscopy so as to determine the pathological damage degree of liver tissue. Results Huoxiao Baifan group could alleviate pathological damage of liver tissue in mice. Conclusion These results suggest that Huoxiao Baifan powder has a markedly protective effect on immune hepatic injury in mice.

Clinical Observation of the Capsule of Xiaokekangs'Effects on Lipids and Blood Glucose to Impaired Glucose Regulation

Geng Jianguo;Li Yunhu;Qi Fang

2007, 28(2):  203-205. 

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Objective To investigate the effects of the capsule Xiaokekang on the impaired glucose regulation(IGR)patients'lipids and blood glucose. Methods 95 impaired glucose regulation patients were divided into treatment group and control guoup randomly and bosseyed, the treatment group took the capsule of Xiaokekang, the control group took placebo, after 6 months treatment, observe the IGR patients'level of serum TC、TG、HDL-C、LDL-C 、FPG、PG2H and the circs of IGR'lapse. Results After treatment, compared to the control group, the treatment group level of serum TC、TG、HDL-C、LDL-C 、FPG、PG2H were declined significantly(P<0.05). Conclusion The capsule of Xiaokekang has the effects of adjusting and reducing the level of lipids and blood glucose, and improving IGR's prognosis and lapse.

Study of the Effects of Shenshuning Mixture on Phenotypic Modulation of Mesangial Cell in Glomerulosclerosis Rats

Zhang Lifen;Huang Wenzheng

2007, 28(2):  206-211. 

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Objective To study the effect of Shenshuning mixture on phenotypic modulation of mesangial cell in glomerulosclerosis rats and its mechanism. Methods Rats models were induced and established by unilateral nephrectomy(Nx) and injection. Rats model of glomerulosclerosis was established by unilateral nephrectomy and intravenous injection of 0.2% Doxorubicin(5 mg/kg), rats were administered Shenshuning mixture 43.93 g/(kg·d), benazepril 10 mg/(kg·d), or distilled water respectively. Other 8 sham-operated rats as comparison. 8 weeks later, The expression of glomerularα-SMA and TGF-β₁ were studied by immunohistochemical staining, and the ratio of positive area/total glomerular capillary area(SMA/GT and TGF/GT)were analyzed with image analysis software. the changes of weight, 24 h proteinuria, plasma albumin, total serum proteim and blood urine nitrogen, creatinine clearance rate were examined in each group. Results Immunohistochemical staining of glomeruli showed that expression of α-SMA and TGF-β₁ in Shenshuning treated group and Benazepril treated group were significantly less than those of model group(P<0.01) and improved rats weights, attenuated 24 h urine protein, blood urine nitrogen and serum creatinine(P<0.01). The relatively analysis showed significant corelation of α-SMA and TGF-β₁ factors(r=0.637, P<0.01). Conclusion Shenshuning mixture can improve glomerulosclerosis by inhibiting phenotypic modulation of mesangial cells and TGF-β₁ expression, attenuate 24 h proteinuria excretion, blood urea nitrogen and creatinine.

Evidence-based Medicine Approach and TCM Therapeutic Evaluation

Liu Jianping

2007, 28(2):  212-215. 

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WHO promotes evidence-based traditional medicine. Methodological issues limit evaluation of therapeutic effect of TCM. Current methods for evaluation do not properly reflect the humanity characteristics and holistic approach and differentiation of syndrome and prescription. TCM therapeutics is a complex intervention, and there is a limitation of methodology by only randomized, double-blind, placebo-controlled trials as it doesn't comply with bio-psycho-social-environmental and spiritual model for modern medicine. To fairly evaluate TCM therapeutic, we need to combine both quantitative and qualitative methods together, so as to create modality and methodology for TCM differentiation of syndrome and treatment.

Research Advances on Mechanism of Nonalcohol Fatty Liver

Liu Shujun;Huang Jingjuan;Che Niancong

2007, 28(2):  216-219. 

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Nonalcoholic fatty liver(NAFL) was the manifestation of the metabolic syndrome. More and more importance has been attached to the high incidence of NAFL. Although the mechanism of nonalcoholic fatty liver is remains undetermined most investigators accept to the second-hit theory.This article has focused on the research advances of insulin resistance in itochondrial dysfunction, cytochromes P450-2E1, and we understand the interaction of multiple factors which induce nonalcoholic fatty liver. It means that teamtherapy will improve the efficacy of clinic treatment.In addition, maybe Traditional Chinese Medicine will be suitable for the therapy because of its advantage of multi-factor integrated treatment. It will have much benefit that using classical Traditional Chinese Medicine to treat nonalcoholic fatty liver on the basis of studying mitochondria, CYP2E1 and insulin.

Hepatotoxicity Caused by Commonly-used Chinese Medicinal Herbs and Compound Preparation

Wang Xiujuan;Xu Liping;Wang Min

2007, 28(2):  220-224. 

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The manifestations of hepatotoxicity caused by Chinese medicinal herbs are similar with ordinary liver disease. Many pathological alterations of hepatotoxicity can be seen, such as hepatocellular damage, liver disfunction, increased serum transaminase, hepatomegalia, jaundice, cholestasis, chronic hepatitis complicated with fibrosis, hepatic cirrhosis、fulminating hepatic failure, liver tumor, and etc. In most cases, the liver damage is reversible after discontinuation. The commonly-seen herbs which may cause hepatotoxicity are Huang Yao Zi(Tuber Dioscoreae Bulbiferae)、Cang Er Zi(Fructus Xanthii Sibirici)、Lei Gong Teng(Radix Tripterygum)、Ku Lian Zi(Fructus Meliae Azedarach), etc., and the compound preparations are Zhuang Gu Guan JIe Wan(The Pill for Asthritis), Ke Yin Wan(The Pill for Psoriasis), Xiao Chai Hu Tang (Xiao Bupleuri Decoction), and so on. In addition, the improper combination of Western medication and Chinese medicinal herbs may also bring about hepatotoxicity. Except the toxicity of herbs themselves, the other factors include the dosage, time, the body constitution of patients, sensitivity, seasons, and geographic regions, etc. Therefore, the medicinal herbs and their preparations should be used rationally under the guidance of the theory of traditional Chinese medicine and differentiation of symptoms, and the medical skill should be improved, which are the effective measures for avoiding hepatotoxicity.

基础研究

The Comparison of Distribution of Dopaminergic Cells and Different Expression of Gene TH and DAT among the Gastrointestinal Tract of Rat

Tian Yuemin;Chen Xin;Wang Wei;Zhu Jinxia

2007, 28(2):  225-228. 

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Objective To study the different expression of dopaminergic cells in different parts of rat gastrointestinal tract. Methods Immunofluorescence was used to study the distribution of DAT in stomach, duodenum, ileum and colon of normal rats. Total RNA was isolated from some of these parts of rats, and subjected to reverse transcription and amplification with TH and DAT specific primers for semi-quantity PCR detection. Results DAT-immunoreactive cells were detected in stomach, duodenum, ileum and colon of normal rats. There were significant differences of TH and DAT mRNA expression in different parts of GI tract of rats. Conclusion Dopaminergic cells are substantially and differently expressed in different parts of rat gastrointestinal tract. They might be involved in the regulation of GI mobility as well as secretion and absorption by dopamine release.

Expression of Human Basic Fibroblast Growth Factor Gene in Baculovirus-Insect Expression System

Wang Yamei;Sun Licui;Yan Yudong;Si Yang;Zhang Jingyi;Qi Yahui

2007, 28(2):  229-233. 

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Objective Bac-to-Bac baculovirus expression system was used to obtain biologically active recombinant human basic fibroblast growth factor(hbFGF) in insect cells. Methods The human bFGF cDNA was cloned into the transfer vector pFastBacHTb. The recombinant plasmid was introduced into E.coli DH10Bac which included a shuttle vector-Bacmid. The cultured Sf21 insect cells were directly transferred with the Bacmid/bFGF DNA, and the pure recombinant baculovirus rAcV-Bac-bFGF was obtained. Then rAcV-Bac-bFGF was infected Sf21 insect cells again to express the hbFGF gene, its expression product was measured by SDS-PAGE and Western blotting. The proliferation level of bFGF was detected by MTT. Results The target protein was detected at the time of 48 h post infection, reached to the peak at the time of 96 h post infection, and persisted expression until 120 h post infection. A 23 000 protein immunostaining band was detected in expressing protein with specific anti-hbFGF antibody by using Western blotting. MTT results showed that rhbFGF proteins had bioactivity to stimulate 3T3 cells proliferation. Conclusion Expression system of Bac-to-Bac baculovirus can be used successfully to express biological activity of hbFGF.

The Effects of Chronic Cerebral Hypoperfusion of the Content of β-amyloid Peptide in Rat's Blood and Brain

Xing Anfeng;Ren Lerong;Ding Ye;Wang Jue;Li Liang

2007, 28(2):  234-238. 

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Objective To investigate the changes of β-amyloid peptide(Aβ) in rats'blood and brain under the condition of chronic cerebral hypoperfusion. Methods Cerebral blood hypoperfusion rat model established by bilateral common carotid artery permanent ligation was used. Learning memory function were examined by Morris water maze. Radioimmunoassay and western blotting were employed to test the levels of Aβ and β-amyloid precursor protein(APP) in serum, cortex and hippocampal. Results The deficiency in learning-memory function began to appear from 1 m after hypoperfusion. Levels of Aβ in serum was significantly increased in each experimental group. Both APP and Aβ were increased from 3 m after hypoperfusion in hippocampal and the same changes occurred from 6m after hypoperfusion in cortex. Conclusion Chronic cerebral hypoperfusion can lead to increase of Aβ both in brain and blood which may play a very important role in early pathogenesis of Alzheimer's disease(AD).

Electrophysiological Properties of Voltage-gated Potassium Channels of Cultured Newborn Bovine Retinal Neurons

Ding Ning;Li Genlin;Wang Jingzhao;Wang Jinjin

2007, 28(2):  239-242. 

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Objective Ion channels are the basis of neuronal signal processing and transmission, and the investigation of the selectivity, kinetics and regulation of varies ion channels on membrane of neurons is of great importance for evaluating the excitability and function of the cells. The characterizations of voltage-activated potassium channels were studied on newborn bovine retinal neurons in long-term cultures with whole-cell patch clamp technique. Methods Neurons from newborn bovine retina were isolated and cultured in vitro. The cells which cultured for 2, 4 or 6 weeks were studied morphologically by phase contrast microscope. Recording pipettes were pulled from borosilicate glass on the two-stage puller. When filled with intracellular solution, the resistances ranged from 2～5 MΩ. The neurons were incubated with standard bathing solution before recording. All voltage-clamp recordings were performed with Axopatch-1D patch-clamp amplifier using tight-seal whole-cell recording configuration under room temperature. Holding potential in the experiment was -70 mV and the test potentials ranged from -190 mV to +90 mV with the increment of 10mV. Whole cell patch clamp recordings were made on 58 neurons from each group. Selective inhibitors of Na+ channels(TTX), Ca²⁺ channels (CaCl₂) and K⁺ channels (TEA, 4-AP, CsCl and BaCl₂) were used to identify the currents. The data were obtained by Clampex software. Current amplitudes and current-voltage relationships were measured and plotted using PClamp 8.0 and Origin 6.0 software. χ² test and one-way ANOVA were used for statistical analysis by SPSS 11.0. Results Cultured neurons isolated from newborn bovine retina could survive in vitro, and had distinct morphological characteristics from co-cultured neuronal glial cells. As the culture went on, the number and length of neurite outgrowth enhanced, indicating the maturity of the cells. With the inhibition of Ca²⁺ and Na⁺ currents by CaCl₂ and TTX, two types of voltage-gated potassium channels were observed. A sustained outward rectifying potassium current was recorded at the depolarized potentials, and the activating threshold was -40 mV. It was suppressed by inducing 30 mmol TEA in the bathing solution. Its kinetics and pharmacology were similar to a delayed rectifier potassium current(I_K) observed in retinal neurons of other species. The average peak amplitude at +90 mV increased as the culture went on [2 weeks: (8.041±126.591) pA, 4 weeks: (549.760±67.673) pA, 6 weeks: (1 697.171±823.456) pA](P<0.05). There was no difference in the expression of I_K among the three groups, and the numbers of cells expressing I_K from each group are 44(2 weeks), 41(4 weeks), and 47(6 weeks) (n=58), respectively(P>0.05). Application of hyperpolarizing steps (-70 to -190 mV) exceeding 200 ms or longer in duration evoked an inward rectifier potassium current which could be partly inhibited by 10 mmol TEA, and be completely inhibited by 5 mmol CsCl or 0.5 mmol BaCl₂. It had a rectifier property as well. Such inward rectifier potassium current has electrophysiological properties identical to I_IR reported from retinal neurons of rat, monkey and human being previously. None of the cells cultured for 2 or 4 weeks expressed I_IR, and only 11 of the total 58 neurons cultured for 6 weeks exhibited such current as well. Conclusion The electrophysiological properties of cultured newborn bovine retinal neurons could be studied using whole-cell patch clamp technique. I_K is commonly expressed, and with the culture went on, the peak amplitude of which increased. Several long-term cultured neurons express an inwardly rectifier potassium current which is assumed to be specifically expressed in primate retinal neurons.

临床研究

A Study of Plasma Tumor Necrosis Factor-alpha in Patients with Gestational Diabetes Mellitus

He Xihong;Lin Li

2007, 28(2):  243-245. 

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Objective Plasma tumor necrosis factor-alpha(TNF-α) in prenatal women with gestational diabetes(GDM) was measured. Methods Fasting plasma TNF-α levels were determined with ELISA in 33 prenatal women with GDM and 53 cases of normal prenatal women, tests repeated for each group according gestational age in four stages of 25～28 weeks, 29～32 weeks, 33～36 weeks and 37～40 weeks. Results ① Plasma TNF-α levels in GDM and normal prenatal group rose as gestational age; ② Plasma TNF-α levels in GDM of four stages were (7.92±1.68)ng/L, (9.46±2.40)ng/L, (10.87±2.38)ng/L, and (11.51±4.55)ng/L respectively, all significantly increased than those in the same stage of normal prenatal group. Conclusion Plasma TNF-α levels in normal prenatal group rise with gestational age. Plasma TNF-α increases more significantly in GDM than in normal pregnancy. Plasma TNF-α is suggested to play an important role in the development of GDM.

Studies of Aphasia after Acute Ischemic Stroke With Proton Magnetic Resonance Spectroscopy

Qu Hui;Fang Ruile;Chen Hongyan;Dong Kehui

2007, 28(2):  246-248. 

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Objective Proton magnetic resonance spectroscopy(MRS) can be used to study metabolite abnormalities in the brains of patients with stroke. We have used it to approach aphasia pathogenesis by observing metabolic changes of language areas on functional magnetic resonance in order to provide theoratic instruction to language rehabilitation of aphasia patients. Methods Between May 2003 and February 2005, 58 in-patients in the Neurology Ward of Beijing Tiantan Hospital, who had been diagnosed with cerebral ischemic by CT or MRI and had language disorders were enrolled for this study. We classified the aphasia types according to by the Western Aphasia Battery, Broca's aphasia and Wernicke's aphasia patients were examin with magnetic resonance spectroscopy(MRS). We measured the Broca's or Wernicke's areas macromolecules and micro molecules, in such as N-acetylaspartate(NAA), choline(Cho), creatine(Cr) and lactate(Lac), and compared the results with that of the mirror hemisphere. Results Of 58 cases, there were 39 male and 19 female. The average age was (64.02±3.87) years. There were 12 Broca's aphasia cases and 21 Wernicke's aphasia cases, rest the other were aphasia types. The results of NAA, Cr, Cho, Lac showed that there were statistically significant differences in the amount of NAA and Cho between Broca's or Wernicke's areas and the mirror hemisphere, but there were no significant differences in Cr. Lactate peak appeared in Broca's or Wernicke's areas, but not appeared in the mirror hemisphere, with significant differences between them. Conclusion Language areas of patients with aphasia are in a state of hypometabolism which may be one of the mechanisms of aphasia.

Survival Analysis on Liver Failure Patients Treated with an Artificial Liver Support System

Wu Wenfang;Zhang Jing;Ma Binrong;Duan Zhongping

2007, 28(2):  249-251. 

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Objective To evaluate the efficacy of artificial liver support system(ALSS) in the treatment of liver failure patients. This is a multi-center, controlled, large sample clinic trial. Methods 916 patients with liver failure from 5 hospitals were studied and followed. All the patients received similar pharmacological manipulation according to one and the same protocol but were divided into an ALSS treatment group and a control group without ALSS treatment. The analysis of survival time was computed using the Kaplan-Maier method, and comparison among groups was done using Log-Rank and Breslow test. Results ALSS has tendency to prolong the survival time; ALSS has obvious effect on survival time of acute liver failure and prolong their survival time; ALSS can affect the short-term survival time of advanced stage patients; ALSS times have an obvious effect on survival time of liver patients especially the acute severe hepatitis and chronic severe ones. Conclusion Multi ALSS treatment is more effective than the standard medicinal liver care treatment. Multi-ALSS treatment could increase survival time of patients suffering from acute liver failure or A and C liver failure. It is important and necessary to treat these patients with ALSS.

Study of Relationship between Hypertension in Patients with Tape-Ⅱ Diabetes Mellitus

Pan Yongyuan;Li Jiaoyuan;Wang Li

2007, 28(2):  252-254. 

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Objective To investigate the relationship between PAC-1 and hypertension in patients with tape-Ⅱ diabetes mellitus(DM). Methods One hundred and twenty eight patients with DM(tape-Ⅱ) were divided into 2 groups: hypertension group(68 cases) and normal blood pressure group(60 cases). Flow-cytometry was used to detect the PAC-1 level, and therefore between the 2 groups was analysed. And the correlation between hypertension, hyperglycemia and PAC-1 was evaluated. Results The PAC-1 level in patients with DM cases was higher than that in control group. It was also markedly higher in patients with hypertension than that in normal blood pressure group. blood pressure and plasma glucose correlated positively with PAC-1. Conclusion Hyperglycemia as sell as hypertension are main factors resulting in a high PAC-1 level.

The Analysis of Necessity and Efficacy of Internal Fixation for Clavicular Fracture

Yang Lixin;Zhang Zezheng;Wang Yuncheng;Wang Hongwei;Zhang Dongzheng

2007, 28(2):  255-257. 

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Objective To investigate the necessity of surgical methods for clavicle fracture and analyze the clinical efficacy of internal fixation. Methods 63 patients with fracture in middle and distal part of clavicle were treated with surgical operation. Thirty fractures from 63 patients were fixed with kirschner's wire. Twenty-eight were treated with reconstructive plate and five with anatomical bone plate. Results All the cases were followed up 12～24 months(means,13months). According to the initial fracture healing standard, 37 cases assessed as excellent, 23 good, 3 poor. The percentage of excellent and good results were 95.24%.The efficacy of internal fixation group(63 cases) was superior to external fixation group(59 cases). Conclusion The internal fixation of clavicular fracture is necessary and the clinical efficacy of internal fixation is excellent. The internal fixation methods of clavicular fracture should be selected according to individual condition.

The Analysis of Ametropia after Penetrating Keratoplasty

Liao Bin;Pan Zhiqiang;Dai Honglei

2007, 28(2):  258-259. 

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Objective To study status of ametropia after penetrating keratoplasty. Methods Optometry was performed for 20 patients with opened pupils three months after penetrating keratoplasty. Results There were 11 eyes of which the visual acuity after penetrating keratoplasty≤0.1, and their corrected visual acuity were 0.05～1.0; there were 9 eyes of which the visual acuity after penetrating keratoplasty≥0.2, and their corrected visual acuity were 0.6～1.2. 15 eyes had astigmatism and 9 eyes were high astigmatism(>3D). 8 eyes were upright axis. There were 7 eyes of which the best corrected visual acuity>1.0 all had the astigmatism, among them 4 eyes were upright axis. Conclusion The corrected visual acuity after penetrating keratoplasty improved significantly. Most cases have high astigmatism with the upright axis. The cause could be that suture are tightened more at 6 and 12, which make tugging to part of cornea, leading to astigmatism with upright axis.

综述

Utility of Gene Chip in Study of Experimental Research inCardiovascular Surgery

Qi Hongwei;Wu Mingying;Tang Chuzhong;Zhu Langbiao;Hu Dayi

2007, 28(2):  262-266. 

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Gene chip technology has successfully been used in the investigation of a diverse array of heart disease as well as other human diseases. It has been used in gene sequence, gene mutation, functional analyses, screening assay, new medicine development and detection of pathogens since 1990. Gene chip technology has been used in cardiovascular disease since 1999. But by now, it has been not used commonly in cardiovascular surgery research.Summarizing the application of gene chip in experimental research of cardiovascular surgery in this paper. Assessing its values and reviewing research progress of cardiovascular surgery. The main ideas of these articles were summarized and analyzed. The successful application of gene chip in the area of cardiovascular surgery revealed lots of unknown things. Gene chip should be used in the area of cardiovascular surgery more and more. It may provide clues and point out directions for the area.