Table of Content

21 October 2015, Volume 36 Issue 5

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Differential expressions of oligomer α-synuclein and phosphorylated α-synuclein in synaptosome in the brain regions of different ages of cynomolgus monkeys of different ages

Yang Weiwei, Li Xin, Li Xuran, Li Xuying, Yu Shun

2015, 36(5):  671-674.  doi:10.3969/j.issn.1006-7795.2015.05.001

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Objective To explore the expressions of oligomer α-synuclein (oligo-α-syn) and phosphorylated α-synuclein (p-α-syn) in synaptosome in brain regions of cynomolgus monkeys of different ages. Methods ELISA was applied to measure the expressions of oligo-α-syn and p-α-syn in different brain regions of cynomolgus monkeys of different ages. Results The synaptosome-α-Syn oligomers and synaptosome-p-α-syn increased significantly in prefrontal lobe, occipital lobe, temporal lobe, hippocampus, striatum and thalamus of old monkeys compared with that of young and middle-aged monkeys (P<0.01). Conclusion The highest level expression of oligo-α-syn and p-α-syn was found in synaptosome of all brain regions in old cynomolgus monkeys.

Expression of α-synuclein in different regions of digestive tract of cynomolgus monkeys

Li Xuran, Yang Weiwei, Li Xin, Yu Shun

2015, 36(5):  675-679.  doi:10.3969/j.issn.1006-7795.2015.05.002

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Objective To investigate the expression of α-synuclein (α-Syn) in different regions of the digestive tract of cynomolgus monkeys. Methods Immunohistochemistry and Western blotting analysis were applied to observe the expression of α-Syn in the digestive tract of cynomolgus monkeys. Results α-Syn immunoreactive substances were observed in both the nerve fibers and nerve plexus in the wall of the digestive tract of cynomolgus monkeys. The α-Syn immunopositive structures were found in the mucosa, submucosal nerve plexus, and myenteric nerve plexus of the digestive tract. The amounts of α-Syn in different regions of the digestive tract were different, with the higher level of α-Syn found in the duodenum, jejunum, ileum, and colon, and lower levels of stomach and esophagus. Conclusion α-Syn is mainly localized in the intrinsic nerves in the digestive tract of cynomolgus monkeys, with the levels differing in different regions of the digestive tract.

Inhibitive effects on MPTP induced alpha-synuclein aggregation by tea polyphenols and EGCG

Yu Lan, Chen Min, Li Xuran, Li Xin, Yang Weiwei, Yu Shun

2015, 36(5):  680-683.  doi:10.3969/j.issn.1006-7795.2015.05.003

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Objective To investigate the inhibitive effects of tea polyphenols (TP) and their extract epigallocatechin-3-gallate (EGCG) on -synuclein aggregation in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-intoxicated Parkinson's disease. MethodsHealthy female 10 to 12 years old cynomolgus monkeys were randomly divided into normal control group (n=4), TP group (n=4), MPTP group (n=4), and MPTP+TP group (n=4). Control group monkeys were not given any treatment; The monkeys in TP group were treated with TP by gastrogavage once daily for 80 consecutive days. In MPTP and MPTP+TP groups, MPTP was injected intravenously as MPTP-HCl to induce PD model. The MPTP+TP groups were treated with TP by gastrogavage once daily for 80 consecutive days after establishing PD model. All animals were euthanized and the brains tissues were rapidly isolated. The level of α-syn oligomers were measured by the ELISA method. In vitro, MES23.5 dopaminergic neurons treated with α-syn monomers or oligomers, followed by treatment with vehicle MPP+ and (or) EGCG. Intracellular levels of α-syn oligomers were measured by ELISA. Cell viability was estimated using the MTT formazan colorimetric assay. Results MPTP-intoxication could increase the levels of α-syn oligomers and TP administration could reduce α-syn oligomers in the brain of MPTP-treated monkeys. In vitro study demonstrated while addition of MPP+ significantly elevated intracellular levels of α-syn in untreated control group and groups treated with α-syn monomers and α-syn oligomers. The capacity of the extract EGCG to reduce -syn oligomerization was further examined and alleviated the MPP+induced cell injury in cultured cells. Conclusion The tea polyphenols and its extract EGCG could alleviate -syn aggregation and dopaminergic neuronal injury caused by MPTP-intoxication.

Preparation and identification of anti-hemoglobin monoclonal antibody

Li Xuying, Li Xin, Li Xuran, Yang Weiwei, Yu Lan, Wang Peng, Yu Shun

2015, 36(5):  684-688.  doi:10.3969/j.issn.1006-7795.2015.05.004

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Objective To prepare a specific antibody against human hemoglobin (Hb) by hybridoma technique, which has great significance in further exploring the functions of Hb in early diagnosis and pathogenesis of Parkinson's disease.Methods The fusion cells were produced with spleen cells of two Balb/c mice immunized with recombinant human Hb to combine with homologous myeloma cells. A positive clone, 5B1, was selected by screening the hybridoma cells with the enzyme-linked immunosorbent assay (ELISA) after three times of limited dilution method. The antibody was acquired by intraperitoneal injection in immunosuppressed BALB/c mice. And Dot blotting, ELISA, Co-immunoprecipitation (Co-IP),Western blotting tests and immunohistochemical (IHC) staining were used for further testing. Results One stable hybridoma cell line, named as 5B1, was found to secret anti-Hb with high titers and specificity. Conclusion The results demonstrated that the monoclonal antibody made in this study has specificity to Hb in human blood, monkey, rats and mouse, and offers a powerful tool for future studies on Hb in Parkinson's disease (PD).

Protective effect of tea polyphenols on dopaminergic neuron in Parkinsonism disease monkeys

Chen Hongwei, Yu Lan, Chen Min, Li Xuran, Li Xin, Yang Weiwei, Wang Peng, Yu Shun

2015, 36(5):  689-693.  doi:10.3969/j.issn.1006-7795.2015.05.005

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Objective To investigate the protecting efficacy of tea polyphenols on dopaminergic neurons in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced Parkinson's disease monkeys and its mechanism. Methods Healthy female cynomolgus monkeys 10 to 12 years old were randomly divided into four groups: normal control group (n= 4), TP group (n=4), MPTP group (n=4), and MPTP + TP group (n=4). Control group were not given any treatment; The TP group monkeys were treated with TP by gastrogavage once daily for 80 consecutive days. In MPTP and MPTP+TP groups, MPTP was injected intravenously as MPTP-HCl to induce PD model. The MPTP+TP groups were treated with TP by gastrogavage once daily for 80 consecutive days after making PD model. Motor deficits were quantified twice a week by a previously validated Parkinsonian monkey clinical rating scale. All animals were euthanized and the brains tissues were rapidly separated. To measure the levels of dopamine and its metabolites in the striatum were measured using HPLC method. Dopaminergic neurons were identified by immunohistochemistry and stereology. Results Tea polyphenols alleviated MPTP-induced injury of nigral dopaminergic neurons and improved motor impairments in cynomolgus monkeys (P<0.05). Conclusion This study showed first in vivo evidence that TP could alleviate dopaminergic neuronal injury and motor impairments in nonhuman primates.

The dynamic changes of reactive oxygen species expression in rat brain tissue following focal cerebral ischemia/reperfusion

Yan Feng, Zhao Yongmei, Luo Yumin, Yin Jie, Li Sen, Fang Yalan, Liu Kejian

2015, 36(5):  694-698.  doi:10.3969/j.issn.1006-7795.2015.05.006

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Objective To investigate the dynamic changes of reactive oxygen species (ROS) during reperfusion time in a rat middle cerebral artery occlusion (MCAO)/reperfusion model and to investigate the role of ROS in cerebral ischemic injury. Methods Twenty-five male Sprague Dawley rats were divided randomly into 5 groups: sham group and MCAO group (90 min ischemia with 3,6,12 or 24 h reperfusion). MCAO operation was performed by using suture method. The rats underwent right MCAO for 90 min. The mean arterial blood pressure (MABP) and rectal temperature were monitored to keep in normal range during the operation. Brains were removed after the animals were euthanized at 3, 6, 12 and 24 h after reperfusion respectively and dyed with 2,3,5-triphenyltetrazolium chloride (TTC) to measure the infarct volume. Cryostat sections were prepared on a freezing microtome. ROS-specific staining and H₂DCF-DA fluorescent staining were used to detect the levels of ROS in the penumbra of the brain. Results 1) There was no statistically significant difference in MABP and rectal temperature between sham and MCAO groups. 2) Sham group rats had no penumbra in the brain where little H₂DCF-DA-positive cells were observed. Lots of H₂DCF-DA-positive staining cells were observed in the penumbra of MCAO group rats at 3, 6, 12 and 24 h after reperfusion. From 3 h after reperfusion, the numbers of H₂DCF-DA-positive staining cells in the penumbra of MCAO group rats showed reperfusion-time-dependent increasing tendency, with significant difference between 3 h and 6 h, as well as 12 h and 24 h (P<0.05). 3) The result of TTC staining showed that no infarction was observed in the brain of sham group rats. The infarct volume of MCAO group rats showed reperfusion-time-dependent increasing tendency (from 3 h to 24 h after reperfusion, P< 0.05). 4) The number of H₂DCF-DA-positively stained cells numbers showed positive correlation with the volume of brain infarction in MCAO group (r=0.833,P<0.01). Conclusion ROS level was increased along with the extension of reperfusion time in a rat focal cerebral ischemia/reperfusion model. ROS might play an important role in cerebral ischemic/reperfusion injury.

Effect of intra-arterial infusion of erythropoietin alone or in combination with tissue plasminogen activator on rats following focal cerebral ischemia

Li Jincheng, Wu Xiaoning, Zhao Haiping, Min Lianqiu, Wang Rongliang, Luo Yumin

2015, 36(5):  699-704.  doi:10.3969/j.issn.1006-7795.2015.05.007

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Objective To investigate the effect of intra-arterial infusion of erythropoietin alone or in combination with tissue plasminogen activator on rats following focal cerebral ischemia/reperfusion, and examine whether the effects are dependent on endoplasmic reticulum stress. Methods Sixty male rats were randomly divided into 5 groups: sham operation (sham), untreated control rats (ischemic animals), erythropoietin(EPO)-treated (onset of reperfusion, 800IU/kg), tissue plasminogen activator (tPA)-treated (onset of reperfusion, 10 mg/kg)and EPO+tPA-treated rats. Middle cerebral artery occlusion (MCAO) was induced by occluding the right middle cerebral artery. Infarct size, neurological deficits, and the levels of phosphorylated form of eukaryotic initiation factor 2α(p-eIF2α) and C/EBP-homologous protein (CHOP) were measured at 24 hours after reperfusion. Results Compared with the control animals, EPO alone (P<0.05) or in combination with tPA (P<0.05) significantly decreased infarct volume, lessened defective degree of neurological function, and reduced expression levels of p-eIF2α and CHOP. tPA+EPO(P<0.05) significantly decreased infarct volume, lessened defective degree of neurological function, and reduced expression levels of p-eIF2α and CHOP when compared with the tPA-treated animals. Immunofluorescent staining revealed co-localization of CHOP and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) in cerebral cortical neurons. Conclusion Based on our data, treatment with EPO or EPO + tPA at the onset of reperfusion by intra-arterial infusion may be considered as a useful candidate to alleviate I/R injury and the potential mechanism underlying the role of EPO in a transient focal cerebral ischemia model in rats may be attributable to suppression of endoplasmic reticulum stress, which preserve neuronal cell viability and attenuate behavioral deficits.

Effect of EX527 on the protection of SH-SY5Y by calorie restriction

Zhang Jingshuang, Ma Lina, Wang Rong, Zhang Jingyan, Wu Yanchuan, Zhao Zhiwei, Wang Yulan

2015, 36(5):  705-708.  doi:10.3969/j.issn.1006-7795.2015.05.008

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Objective To observe the impact of SIRT1 inhibitor (EX527) on the human neuroblastoma cell line SH-SY5Y, and the SH-SY5Y cells were treated with calorie restriction. Methods SH-SY5Y cells were cultured in vitro and divided into the following three groups: normal control group (NC group), calorie restriction group (CR group) and calorie restriction plus SIRT1 inhibitor EX527 group (CR+EX527 group). The morphological changes in the SH-SY5Y cells of each group were observed by light microscopy, and the thiazolyl blue tetrazolium bromide (MTT) metabolic rate and the lactate dehydrogenase (LDH) leakage rate of all groups were measured by MTT working liquid and LDH Cytotoxicity Assay Kit, and the expression of SIRT1 protein in the three groups mentioned above were detected by Western blotting. Results Compared with the normal control (NC) group, cells of CR group were in good shape, including plump cell bodies, good adherence and neurites extension. The MTT metabolic rate of CR group increased (P<0.05), while the LDH leakage rate tended to decrease as compared with NC group. And the expression of SIRT1 protein of CR group and CR+EX527 group were increased obviously (P<0.05) as compared with NC group. Compared with the calorie restriction (CR) group, cells of CR+EX527 group showed poor morphology, including no plump cell bodies, retracted neurites and bad adherence. The MTT metabolic rate of CR+EX527 group decreased (P<0.05), while the LDH leakage rate tended to increase as compared with CR group. And the expression of SIRT1 of CR+EX527 group were decreased slightly as compared with CR group. Conclusion Calorie restriction has a protective effect on nerve cells, and SIRT1 inhibitor EX527 can weaken the neuroprotective effect of calorie restriction in this study, so we preliminarily infer that the protective effect of calorie restriction on SH-SY5Y cells may be mediated by SIRT1.

Effects of normobaric hyperoxia on activation of astrocyte following cerebral ischemia-reperfusion in rats

Shi Wenjuan, Liang Jia, Dong Wen, Qi Zhifeng, Liu Kejian

2015, 36(5):  709-713.  doi:10.3969/j.issn.1006-7795.2015.05.009

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Objective To observe the effect of normobaric hyperoxia (NBO) on GFAP (the activation marker of astrocyte) and connexin43 expression induced by cerebral ischemia-reperfusion injury in rats, and preliminarily explore the effect of NBO treatment on activation of astrocytes. Methods A total of 15 healthy adult male Sprague-Dawley (SD) rats (280-320 g) were randomly divided into Sham group (n=3), normoxia group (n=6) or NBO group (n=6). A model of middle cerebral artery occlusion for 1.5 h and reperfusion for 24 h was induced by using the intraluminal suture method. The sham group and the normoxia group rats breathed normal air, and instead NBO group rats were exposed to 100% oxygen after ischemia from 5 minutes until reperfusion. Immunofluorescence staining and Western blot were used to observe the expression of glial fibrillary acidic protein and connexin 43 in basal ganglia and cortex after cerebral ischemia.Results Immunofluorescence staining results showed that compared with the normoxia group, the number of GFAP positive cells in the ischemic cortex of NBO group was significantly reduced with the processes and fluorescence intensity decreased. Western blot results showed that compared with the normoxia group, the expression of GFAP protein was decreased in NBO group, and GFAP expression in the ischemic cortex of NBO group was statistically significantly reduced (P<0.05), and it was consistent with the immunofluorescence staining results. Compared with the sham group, the Cx43 protein expression was decreased in the normoxia group and that in the ischemic cortex of the normoxia group was statistically significantly reduced (P<0.01). Compared with the normoxia group, the Cx43 protein expression was increased in NBO group and that in the ischemic cortex of NBO group was statistically significantly increased (P<0.01). The Cx43 protein expression in basal ganglia did not change significantly. Conclusion NBO treatment decreased the excessive activation of astrocyte probably through increasing Cx43, therefore, NBO may play a protective role in the ischemia-reperfusion injured brain.

Role of CD62p in the diagnosis of acute brain infarction disease

Fu Lu, Wang Zhaohui

2015, 36(5):  714-717.  doi:10.3969/j.issn.1006-7795.2015.05.010

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Objective To explore the relationship between the expression rate of platelet membrane glycoprotein CD62p and acute cerebral infarction, draw ROC curve, determine the cutoff value, and clarify its clinical significance. Methods Flow cytometry that adopts FITC and PE double calibration was used to detect CD62p positive rates in the plasma of acute cerebral infarction group patients, then comparative analyses were performed between the high-risk group and the control group. The relationship between the CD62p and cerebral infarction was analyzed by drawing the ROC curve and it was confirmed that the CD62p positive rate was applicable to determine the best cutoff value of acute cerebral infarction. Results Acute cerebral infarction group had a significantly higher positive CD62p expression rate than the high risk group and the control group, which are all statistically significant (P<0.01). Dynamic observation of the CD62p positive expression rate of acute cerebral infarction group showed that plasma CD62p expression rate was the highest in 24 h, a trend of gradual decline on 7 d and 14 d, but remained higher than that of the high risk group and the control group on 14 d. The results are statistically significant (P<0.01). ROC curve drown by using CD62p positive expression rate showed that 5.20% was the cutoff point. The sensitivity was 92%, specificity 95%, positive predictive value 98%, negative predictive value of 73% and coincidence rate (accuracy) 92%. Conclusion In the patients of both the acute cerebral infraction group and high risk group, the platelet was kept high level, but the platelet activation in acute cerebral infarction group was higher, and in turn it can be speculated that the higher the positive expression rate of CD62p, the serious the patient,s condition. When the detected CD62p is higher than 5.2% after analyzing the ROC curve which is drawn based on CD62p positive expression, it indicates very high possibility of cerebral infraction. Therefore, measuring CD62p could predict the occurrence of acute cerebral infarction.

Protective effect and the related mechanisms of rhubarb on ischemic cerebrovascular disease

Fang Yalan, Luo Yumin, Zhao Yongmei

2015, 36(5):  718-722.  doi:10.3969/j.issn.1006-7795.2015.05.011

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Cerebral ischemia is the main leading cause of death and adult disability. Early thrombolysis is the most effective way to contain cerebral ischemic injury, however, its narrow therapeutic window and association with an increased risk of intracranial hemorrhage limits its application. As a traditional Chinese herbal medicine, rhubarb has many kinds of biological activities, such as anti-inflammatory, anti-oxidant, and anti-apoptotic effects. This paper reviews the neuroprotective effect and the related mechanisms of rhubarb on ischemic cerebrovascular disease.

The effect and mechanism of glyburide on monocrotaline-induced pulmonary hypertension in rats

Liu Ting, Liu Yafei, Wang Wang, Chai Sanbao, Liu Jie, Wang Jun

2015, 36(5):  723-728.  doi:10.3969/j.issn.1006-7795.2015.05.012

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Objective To investigate whether glyburide could prevent monocrotaline (MCT)-induced pulmonary hypertension (PH) via inhibiting nucleotide-binding domain and leucine-rich repeat PYD-containing protein 3 (NLRP3) inflammasome activation in rats. Methods Male Sprague-Dawley (SD) rats were divided into 3 groups: control group, MCT group and MCT+glyburide group. Hemodynamic changes, right ventricular hypertrophy, and lung morphological features were assessed. The levels of NLRP3 and interleukin-1β(IL-1β) were assessed by Western blotting and ELISA. Results Compared with control group, four weeks after MCT injection, the right ventricular systolic pressure (RVSP), the index of right ventricular hypertrophy (RVHI) and the percentage of medial thickness (MT%) of pulmonary artery significantly increased by 1.66 (P<0.001), 2.15(P<0.001) and 1.20 folds (P<0.001), respectively. Compared with MCT group, they reduced by (33.60±5.14)% (P<0.05), (18.26±9.23)% (P<0.05) and (21.86±8.64)% (P<0.001) in MCT+glyburide group. Compared with control group, the expression of NLRP3 in lung of MCT group increased by 1.57 folds (P<0.05), while it decreased by (55.96±19.33)% (P<0.01) after using glyburide in 4 weeks. The serum level of IL-1β was higher in MCT group than that in control group[ (1 915.82±316.47)pg/mL vs (118.29±27.60)pg/mL, P<0.001], while it was lower in MCT+glyburide group than that in MCT group [(396.64±237.19)pg/mL vs (1 915.82±316.47)pg/mL, P<0.001]. Conclusion These results suggested that glyburide could ameliorate MCT-induced pulmonary hypertension that was mediated by the NLRP3/IL-1β pathway.

Regulation of inflammatory cytokine expression of sphingosine 1-phosphate in murine monocyte/macrophage J774A.1

Zhang Yuanyuan, Li Weiyang, Yang Lin, Li Liying

2015, 36(5):  729-733.  doi:10.3969/j.issn.1006-7795.2015.05.013

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Objective The aim of study is to explore the role of sphingosine 1-phosphate (S1P) signaling on murine monocyte/macrophage inflammatory cytokine expression. Methods Western blotting was employed to test whether S1P receptors (S1PRs) expression in murine monocyte/macrophages. Then we detected whether S1P-mediated expression of inflammatory cytokines in murine monocyte/macrophage J774A.1 by real-time PCR. We studied regulatory mechanisms of expression of inflammatory cytokines in J774A.1 cells by intervention of S1P/S1PRs signaling with pharmacology tools. Results S1PR₂ antagonist JTE-013, S1PR₃ antagonist CAY-10444 treatment reduced expression of inflammatory cytokines in J774A.1 cells. S1PR₁ antagonist W146 has no effects on the expression of inflammatory cytokines in J774A.1 cells. Conclusion S1P up-regulates expression of inflammatory cytokines in J774A.1 cells mediated by S1PR₂ and S1PR₃.

L-DOPA regulates MAO-B transcription through DNA methylation in SH-SY5Y cells

Yang Zhaofei, Wang Yong, Yang Jian, Wang Xuan, Wang Xiaomin

2015, 36(5):  734-739.  doi:10.3969/j.issn.1006-7795.2015.05.014

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Objective To explore the effect and molecular mechanism of L-3,4-dihydroxyphenylalanine (L-DOPA) on transcription of monoamine oxidase B (MAO-B), which was a key gene in dopamine metabolisms. Methods SH-SY5Y cells were used as the object to mimic dopaminergic neurons in Parkinson's disease. Methylated DNA quantitation kit was used to analyze genomic 5-mC content. The gene transcriptional levels were measured by semi-quantitative RT-PCR. Methylation specific PCR (MSP) was applied to detect the promotor CpG methylation status of MAO-B. Results 1 Genomic 5-mC content was decreased in SH-SY5Y cells treated with L-DOPA (2 μmol/L and 20 μmol/L) for 24 h. 2 Monoamine oxidase B (MAO-B) mRNA level was significantly down-regulation in L-dopa administration, but neither DOPA-decarboxylase (DDC) nor catechol-O-methyltransferase (COMT). 3 A DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5-aza-dC), induced MAO-B mRNA level up-regulation, but COMT and DDC mRNA levels did not affected. 4 CpG hyper-methylation of MAO-B was induced by L-DOPA (2 μmol/L and 20 μmol/L) for 24 h. And CpG hypo-methylation was induced by 5-aza-dC. Conclusion Transcription of MAO-B was down-regulated by L-DOPA and might be mediated by L-DOPA induced promoter CpG hyper-methylation. This study might provide a new evidence in the view of epigenetics, especially DNA methylation, on dopamine dysregulation and L-DOPA induced side effects in clinic.

Parkinson's disease related gene PINK1 but not DJ-1 was down-regulated in neurotoxicity of 6-hydroxydopamine and 1-methyl-4-phenylpyridinium

Yang Jian, Wang Yong, Wang Xuan, Yang Zhaofei, Gong Xiaoli, Wang Xiaomin

2015, 36(5):  740-746.  doi:10.3969/j.issn.1006-7795.2015.05.015

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Objective To explore the transcriptional levels and molecular mechanism of key Parkinson's disease (PD)-related genes DJ-1 and PINK1 in neurotoxicity. Methods MN9D cells were used as dopaminergic neurons and treated with neurotoxins 6-hydroxydopamine (6-OHDA) and 1-methyl-4-phenylpyridinium (MPP⁺). Lethal doses 50 (LD 50) was determined by cell counting with Typan Blue staining. Transcriptional levels of DJ-1, PINK1 and demethylation enzyme TET1 were detected by semi-quantitative RT-PCR. Expressional levels of demethylation enzyme TET1 were detected by immune-cytofluorescent staining. Results LD50 at 10 μmol/L of 6-OHDA and 100 μmol/L of MPP⁺ were used to treat MN9D cells. In neurotoxicity of 6-OHDA and MPP⁺, transcriptional levels of PINK1 were significantly down-regulated, but no change in DJ-1 transcriptional levels was found. In addition, transcriptional and expressional levels of TET1 were down-regulated in neurotoxicity of 6-OHDA and MPP⁺. Conclusion These data suggest that down-regulated TET1 might contribute to the down-regulation of PINK1 in neurotoxicity of PD.

Effects of tangshenning on HGF expression of renal tissue in type 2 diabetic KK-Ay mice

Liu Yingxin, Zou Dawei, Geng Jianguo, Gao Yanbin, Shang Yawen, Li Jiaoyang, Gong Muxin, Peng Qizhen

2015, 36(5):  747-750.  doi:10.3969/j.issn.1006-7795.2015.05.016

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Objective To study the effects of Tangshenning on the protein expression of Hepatocyte Growth Factor (HGF) in KK-Ay mice with spontaneous type 2 diabetic mellitus(T2DM). Methods Diabetic male KK-Ay mice were randomly divided into three groups: the model group, the valsartan group and the Tangshenning group, with 20 in each. Twenty male C57BL/6J mice were chosen as the normal group. After intervention with Tangshenning and valsartan for 12 weeks, hemoglobin A1c (HbA1c), blood urea nitrogen (BUN)and serum creatinine (SCr) were examined. Expression of HGF was assessed by Western blotting. Results Compared with blank group, HbA1c, BUN and SCr in model group are significantly increased (P<0.05) while the expression of HGF was significantly decreased. Compared with model group, there were no significant changes in the levels of HbA1c (P>0.05). The mean levels of BUN and SCr in both treatment group were lower than model group (P<0.05). The expression of HGF was significantly increased. Conclusion Tangshenning can decrease BUN, SCr and prevent DN by promoting the expression of HGF.

Study on the preservation with carbon oxide and heterotopic transplantation of the isolated rabbit heart

Guo Yilong, Zhang Zhong, Zhou Pengyu, Zhu Ping, Huang Shuai, Zheng Shaoyi

2015, 36(5):  751-756.  doi:10.3969/j.issn.1006-7795.2015.05.017

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Objective Investigating the protective effect and mechanism of hypothermia (4 ℃)and desiccation with CO(CO=0.8ATA,O₂=3.2ATA,1ATA=760 mmHg, 1 mmHg=0.133 kPa)on heart preservation. Methods Seventy-two rabbits were used in this experiment; They were divided into 5 groups randomly. Group Ⅰ(eight of them were collected for examination without transplantation or preservation);Group Ⅱ(sixteen of them were divided into this group and the isolated hearts were underwent abdominal heterotopic transplantation without preservation);Group Ⅲ(sixteen of them were divided into this group and the isolated hearts were preserved with desiccation);Group Ⅳ(sixteen of them were divided into this group and the isolated hearts were preserved with desiccation and CO);Group Ⅴ(sixteen of them were divided into this group and the isolate hearts underwent 18 h preserved with HTK solution); Group Ⅲ-Ⅴ were placed in a refrigerator at 4 ℃; And then, the isolated hearts were underwent abdominal heterotopic transplantation. After the heterotopic transplantation, the re-beat rates were counted; 1 h after transplantation, the blood of receptor and the myocardium of donor were collected to measure the enzyme of function and the inflammatory factors of myocardium; Results After the experiment, most of the enzyme of function (CK-MB: creatine kinase-MB; CK: creatine kinase; LDH: lactic dehydrogenase) and the inflammatory factors of myocardium (TNF-α: tumour necrosis factor-α;IL-1: interleukin-1;IL-17: interleukin-17) in Group Ⅲ were lower than those of Group IV (P<0.05); Conclusion The heart preserved with CO was significantly better than that preserved with traditional method. And this new preservation works by reducing the inflammatory factors and alleviating the myocardia reperfusion injury.

Monitoring synthesis systems of acetylsalicylic acid with citric acid as the catalyst by second derivative spectrometry

Shao Jianqun, Xu Yanxia, Tang Jingcheng, He Shenzhi, Zhang Feng

2015, 36(5):  757-760.  doi:10.3969/j.issn.1006-7795.2015.05.018

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Objective To establish a method for monitoring synthesis systems of acetylsalicylic acid with citric acid as the catalyst by second derivative spectrometry.Methods Respectively, standard work curve of acetyl salicylic acid and salicylic acid were established with second derivative value against concentration at the wavelengths of 289nm and 308nm. Under different temperatures, the real-time content of acetyl salicylic acid and salicylic acid in synthesis reaction system were monitored. Results When the reaction temperature is 70 ℃,80 ℃,90 ℃ and 95 ℃, the complete reaction times were about 80 min, 50 min, 20 min and 15 min individually. Conclusion The second derivative spectrometry method can be achieved on the monitoring and judging acetylsalicylic acid synthesis reaction endpoint.

Pathological morphology change of respiratory and peripheral immune organ and the change of immune cells

Qi Jing, Lyu Fudong, Duan Xuejing, Wang Jue, Wang Daye, Xu Xin, Bai Yunfei, Li Hongjun, Jin Ronghua, Li Ning, Dai Jie

2015, 36(5):  761-767.  doi:10.3969/j.issn.1006-7795.2015.05.019

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Objective To explore the pathology morphological changes of trachea and peripheral immune organ and immune cells of the new influenza A (H1N1) virus severe infection. Methods To collect Collection of 2009 Beijing influenza A H1N1 intensive deaths 8 cases; Pathological changes of these tissue were observed by HE staining; The immune cell types were tested, quantity and distribution characteristics were made by immunohistochemistry. Results The principal pathologic changes of trachea were necrotizing bronchiolitis with peripheral inflammation, diffuse alveolar damage, pulmonary hemorrhage and fibrosis. Macrophages hyperplasia and little lymphocyte infiltration; Viral antigen was found in the epithelium of the epithelium and macrophage; CD68⁺ macrophage count significantly increased in lung, CD20⁺ B lymphocytes, CD3⁺, CD4⁺, CD8⁺ T count, CD56⁺ NK cells occasional,no obvious differences were found compared with the control group (P>0.05). Spleen and lymph nodes were same,focal histocyte hyperplasia, erythrocytophagy were observed, lymphoid tissue atrophy.IHC cell count showed CD68⁺ macrophage significantly increased, CD20⁺ B, CD3⁺, CD4⁺, CD8⁺ T lymphocytes were significantly lower than the control group (P<0.05). NK cells were rarely seen in the spleen and lymph nodes. The ratio of CD4⁺ and CD8⁺ T lymphocytes between experimental group and control group is not significant different. Conclusion The severe influenza A (H1N1) patients, peripheral immune organs showed significant atrophy, specific immune function was abated and the cellular immune response decreased more significantly.

Effectiveness of cognitive behavioral intervention on the maintenance of chronic periodontitis, treatment

Ding Fang, Lyu Yalin, Wang Jie, Liu Yinhua

2015, 36(5):  768-772.  doi:10.3969/j.issn.1006-7795.2015.05.020

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Objective To observe if cognitive behavioral intervention could maintain the treatment effect for chronic periodontitis. Methods Eighty five patients with chronic periodontitis were randomly divided into two groups. The controls accepted routine oral hygiene instruction after periodontal initial therapy. The experimental group accepted cognitive behavioral intervention including oral health education. The periodontal clinical index was noted at baseline, 2 months, 4 months and 6 months after periodontal initial therapy. Results The plague index (PLI), bleeding index (BI), probing depth (PD) and attachment loss (AL) of the experimental group were lower than those of the controls at 2 months after treatment, though the difference was not significant. The PLI of the experimental group at 4 months after treatment was significantly lower than that of the controls(1.48±0.29 vs 1.77±0.31, P =0.000). Also, the BI of experiment group was significantly lower that of than the controls (2.41±0.31 vs 2.56±0.23, P=0.013), the PD of experimental group was significantly lower than that of the controls[(3.18±0.25)mm vs (3.38±0.30)mm, P=0.001]. The AL of experimental group was also significantly lower than that of the controls at 4 months after treatment, (2.59±0.21 vs 2.77±0.30, P=0.002). At 6 months after treatment, the PLI、BI、PD and AL of the experiment group were significantly lower than those of the control group. Conclusion The effectiveness of cognitive behavioral intervention on the maintenance of periodontal initial treatment for periodontitis is significant. And it can belong to the category of periodontal maintenance therapy.

Effect of early blood pressure variability on the prognosis of patients with severe cerebral hemorrhage

Ren Tianhua, Yang Tiecheng, Li Jianguo, Niu Chi, Li Lixia, Shi Hongmei

2015, 36(5):  773-777.  doi:10.3969/j.issn.1006-7795.2015.05.021

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Objective To investigate the association between early blood pressure variability and prognosis in patients with severe cerebral hemorrhage. Methods A retrospective analysis of 808 patients with severe cerebral hemorrhage treated at Emergency Department Intensive Care Unit of Beijing Tiantan Hospital from 2009 to 2011 was performed. According to the design of this study,108 cases of inclusion criteria patients were selected randomly. According to the 7-day outcome after admission to EICU, the patients were divided into nonsurvivors(51 cases)and survivors (56 cases), and the blood pressure level in them was monitored in the first 24 hours. Initial systolic blood pressure(ISBP)、mean systolic blood pressure(MSBP)、mean diastolic blood pressure(MDBP)、blood pressure variability(BPV)、smoothing index(SI), the Glasgow coma scale (GCS) at admission、nicardipine total dosage in 24 hours. ResultsGCS、ISBP、MSBP、MDBP、DBPV showed no significant differences between nonsurvivors(51 cases)and survivors(6.38 ±1.74) vs (6.45±1.39)、 (201.6±16.32)mmHg vs (195.5±13.47)mmHg、(155.2±11.6)mmHg vs (153.1±10.8)mmHg、(88.5±8.2)mmHg vs (85.38±7.4)mmHg、(9.32±2.17)mmHg vs (8.91±2.58)mmHg,all P>0.05(1 mmHg=0.133 kPa),but SBPV and nicardipine total dosage (NTD) were significantly higher in nonsurvivors than survivors(14.56±6.87) vs (11.30±2.48)、(105.24±19.66)mmHg vs (78.57±11.25)mmHg,all P<0.05, smoothing index of systolic blood pressure (SISBP) and smoothing index of diastolic blood pressure (SIDBP) were significantly lower in nonsurvivors than survivors (1.125±0.124) vs (1.464±0.158)、(1.147±0.231) vs (1.263±0.245),(P<0.01,P<0.05). Conclusion The early blood pressure variability was closely related to early outcome of patients with severe cerebral hemorrhage,control early blood pressure fluctuation might improve patients, outcome.

Study on the relationship between the nonalcoholic fatty liver and serum iron and ferritin

Zhang Wenping, Wu Jing, Ren Meijie, Chen Yanxing, Li Kechun, Yang Peiyi, Yang Jiaxnwei, Wang Mengnan, Nie Meiling

2015, 36(5):  778-781.  doi:10.3969/j.issn.1006-7795.2015.05.022

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Objective To investigate the relationship between the nonalcoholic fatty liver disease (NAFLD) diagnosed by ultrasonography and serum iron and ferritin. Methods Forty patients with NAFLD (case group) and forty normal persons(control group) were examined by ultrasonic diagnosis, the levels of serum iron and serum ferritin were detected by atomic absorption spectrum method and radioimmunoassay respectively. Results The level of serum iron and serum ferritin in case group were (105.17±24.52)μg/dL,(146.57±68.28)ng/dL, the level of serum iron and serum ferritin in control group were (85.45±26.84)μg/dL,(90.58±49.71)ng/dL, respectively.The difference in serum iron and serum ferritin was significantly different between NFALD patients and normal control (P<0.01). The area under receiver operating characteristic (ROC) curve diagnosing NAFLD by serum iron and the best threshold value and its sensitivity,specificity,mistake diagnosis rate,omission diagnosis rate are 0.69(P<0.01),123.5 (μg/dL),82.5%,60%,40 %,17.5% respectively. The area under ROC curve diagnosing NAFLD by serum ferritin and the best threshold value and its sensitivity,specificity,mistake diagnosis rate,omission diagnosis rate are 0.73(P<0.01),134.9 (ng/dL),75.0%,72.5%,27.5%,25.0% respectively. Conclusion The levels of serum iron and serum ferritin were higher in NAFLD patients. The rise of serum iron and serum ferritin is coincide with NAFLD diagnosed by ultrasonography.

Clinical significance of joint detection of serum homocysteine, lipoprotein(a) and high sensitive C-reactive protein in cerebral infarction

Wang liqiu, Yin changbin, Li jiawei

2015, 36(5):  782-785.  doi:10.3969/j.issn.1006-7795.2015.05.023

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Objective To investigate the changes and the clinical significance of joint detection of serum homocysteine (HCY), lipoprotein(a)[Lp(a)] and high sensitive C-reactive protein (hs-CRP) in patients with cerebral infarction. The cut of value of three results of the patients with cerebral infarction was determined by ROC curve. Methods The levels of HCY, Lp(a),hs-CRP were measured in 183 patients with cerebral infarction and 163 healthy controls and the results were made statistical analysis. Results The levels of Hcy,Lp(a) and hs-CRP in patients with cerebral infarction were (16.7±6.3) μmol/L, (343.6±150.1)mg/L, (3.87±1.88)mg/L, in healthy control group were (12.9±4.7) μmol/L, (205.4±190)mg/L,(2.45±0.69)mg/L. The levels of Hcy,Lp(a) and hs-CRP in patients with cerebral infarction were significantly higher than those in healthy controls(P<0.05).Conclusion Occurrence of cerebral infarction and Hcy, Lp(a) and hs-CRP levels were closely correlated,so the indexes can be used as risk factors.

Treatment of high grade glioma with VM-26 and DDP in 30 cases

Li Yan, Shi Rui, Chen Jianxin, Li Shan, Kang Xun, Cheng Jing, Zhao Kaihong, Li Wenbin

2015, 36(5):  786-790.  doi:10.3969/j.issn.1006-7795.2015.05.024

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Objective To evaluate the safety and efficacy of teniposide (VM-26) and cisplatin (DDP) in the treatment of recurrent malignant glioma. Methods A total of 30 patients were included for this study. The patients were diagnosed with recurrent glioma between 2012/05/01 and 2014/4/30. All patients experience classical STUPP project which contains surgery resection and adjuvant radiotherapy and chemotherapy. The diagnoses were confirmed by the clinical symptoms, contrast-enhanced MRI and the consensus on histology from at least two neuropathologists at the Department of Oncology in Beijing Shijitan Hospital. After the treatment, the image based progression and the adverse events rates were applied to evaluate the results. Results The efficiency for grade Ⅲ glioma patients was 9.1%. The survival rate was 43 months. At the meanwhile, the efficiency for grade Ⅳ was 5.3% and the survival rate was 16 months. The mean overall survival rate of 30 high-grade glioma patients was 18 months. Conclusion The VM-26 and DDP are potential therapeutic agents to treat patients with high-grade malignant recurrent glioma. And VM-26 and DDP have an acceptable safety profile and its adverse events are mostly mild.

Analgesic efficacy in different approach of ultrasound-guided transversus abdominis plane block after cesarean delivery

Xie Hai, Li Yan

2015, 36(5):  791-793.  doi:10.3969/j.issn.1006-7795.2015.05.025

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Objective To compare the analgesic efficacy of anterior axillary line approach with subcostal approach in ultrasound-guided transversus abdominis plane block after cesarean delivery. Methods Singlebirth primigravida 144 cases undergone longitudinal incision cesarean section with single spinal anesthesia were randomly divided into group A (anterior axillary line approach group) and group B (subcostal approach group)(n= 72). All cases in the two groups were done via ultrasound guided transversus abdominis plane block after surgery; 30 mL of 0.25% ropivacaine were injected in each side after ultrasound to determine the transversus abdominis plane. Visual analogue pain (VAS) score in postoperative 6, 12, 24, 48 h time points, analgesic intensity peak time, postoperative ambulation time and patient satisfaction with postoperative analgesia in the two groups were observed. Results Visual analog pain (VAS) score in postoperative 6, 12, 24, 48 h time point between the two groups was not statistically significant difference (P> 0.05). Time of analgesic intensity peak time in group A was (2.9±1.8) h and group B (2.6±1.4) h with no significant difference (P> 0.05). The average time to get out of bed in the two groups showed no significant difference (P> 0.05). Patient satisfaction with postoperative analgesia in the two groups was no significant difference (P> 0.05).Conclusion Postoperative analgesia in cesarean section with longitudinal incision via either anterior axillary line approach or subcostal approach in ultrasound-guided transversus abdominis plane block is available within 24 hours, both approaches are without difference.

Molecular classification of brain tumors guides individualized treatment

Gao Hua, Zhao Peng, Zhang Yazhuo

2015, 36(5):  794-799.  doi:10.3969/j.issn.1006-7795.2015.05.026

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Primary brain tumors are among the most lethal of all cancers, largely as a result of their lack of responsiveness to current therapy. Numerous new therapies hold great promise for the treatment of patients with brain cancer, but the main challenge is to determine which treatment is most likely to benefit an individual patient. By reviewing advances of brain tumors,we suggest that a more biological approach to brain tumors classification will provide improved means to type these tumors. Any new classification, however, must be based on clinical significance, and we thus point out the pressing need for better clinical endpoints and outcome measures in the field.

Translational medicine and the development of minimally invasive surgery in China

Li Jun, Guo Wei, Zhang Jun, Zhang Zhongtao

2015, 36(5):  800-804.  doi:10.3969/j.issn.1006-7795.2015.05.027

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Translational medicine (TM) was first introduced in 1992. The core objective of TM is to translate basic scientific findings into therapeutic interventions for patients. TM can increase the understanding of important disease processes. In recent years, the investment in basic medical research has been increasing, but very few results can benefit clinical practice. To prevent basic and clinical medicine being disjointed and fragmented, TM is urgently needed. In worldwide, the development history of minimally invasive surgery (MIS) is also the process of collaborative development between basic medicine and clinical medicine. China adopted the minimally invasive surgery later than the western countries. However, through the early TM efforts of Chinese academics, Chinese MIS is building up a momentum and starts to impact MIS field worldwide. The most important issue at this time is how we improve the conversion rate of basic medicine to clinical medicine. Here, we briefly review the applications and difficulties of TM in MIS, especially in China.

Translational study of electrical therapy for spinal cord injury from basic research to clinical technology

Lyu Zhen, Jiang Shudong, Bai Jinzhu, Zhang Junwei, Wang Fangyong, Hong Yi

2015, 36(5):  805-808.  doi:10.3969/j.issn.1006-7795.2015.05.028

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Objective Electrical therapy is an important treatment in the clinical spinal cord injury. It provides an effective modality to improve the various dysfunction after spinal cord injury (SCI). Methods Four kinds of electrical technologies were reviewed. They are electrical stimulation on spinal cord injury site, functional electrical stimulation on the paralysis of limbs, diaphragm pacing on respiratory insufficiency and neural modulation on voiding dysfunction. Results (1)DC electric field could promote untransected axonal regeneration, and the oscillating electric field can promote two-way repair of the cranial and caudal spinal axons. (2)Functional electrical stimulation, used to reconstruct the neuromuscular function, stimulate the muscle of the limb, causing muscle contraction and limb motion, thus complete the extension of elbow, grasping, standing, walking and other motor function, and prevent complications.(3)Diaphragm pacing can induce diaphragm muscle contraction, to maintain the natural negative pressure breathing in patients with diaphragmatic dysfunction.(4)The electric frequency of 5-20 Hz and pulse width 1-5 ms can both effectively inhibit detrusor contraction and improve pelvic functions. Conclusion Electric therapy as a treatment technology, shows not only the similar effects as bioelectricity, but also various characteristics under different electrical conditions. In future research, the combination of electrical treatment technology and other technology should be strengthened, so as to solve more clinical problems.

Review of mechanisms of acupuncture in treatment of Alzheimer's disease

Li Hui, Qi Zhigang, Li Kuncheng

2015, 36(5):  809-813.  doi:10.3969/j.issn.1006-7795.2015.05.029

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Alzheimer's disease(AD) is a progressive neurodegenerative disorder characterized by cognitive impairment. Acupuncture, an important component of Traditional Chinese Medicine, has been used in treatment of dementia for many years, but the mechanism is still unclear. In the past few decades, researchers in manyparts of the world have devoted to studying the essence of acupuncture treatment on AD. Conclusion drawn from experiments included that acupuncture plays a protective role through inhibiting harmful factors and promoting the beneficial ones. On the other side, using fMRI, researchers concluded that acupuncture could evoke activation or deactivation and modulate the functional connectivity between distinct brain regions, these effect of acupuncture on brain would be another important mechanism.

Application of contrast-enhanced ultrasound in cerebrovascular disease

Wu Dongfang, He Wen, Shi Chuanying, Duan Fengxia

2015, 36(5):  814-819.  doi:10.3969/j.issn.1006-7795.2015.05.030

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Over the last few years, Ultrasound is readily available, cheap, safe, lacks ionizing radiation and accurately identifies the presence of structural abnormalities. Nevertheless, ultrasound, including Doppler ultrasound,these methods have some limitations,including failing to accurately identify and detect intraplaque neovascularization and intracranial aneurysms and so on.Recently,Contrast-enhanced ultrasound (CEUS) has been introduced as a technique that plays an important role in the assessment of to improving the detection of blood in the vessel and intraplaque neovascularization. CE-TCCS can improve the sensitivity and detection rate of intracranial aneurysms and more clear display of intracerebral hematoma location,shape,boundary, provide blood perfusion information of surrounding tissue in hematoma.

Mechanisms of DNA repair for the maintenance of genome stability in cells——introduction of 2015 Nobel Prize in Chemistry

Kong Lu, Ding Wei

2015, 36(5):  823-828.  doi:10.3969/j.issn.1006-7795.2015.05.032

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The 2015 Nobel Prize in Chemistry was honored to three scientists of Tomas Lindahl from Sweden, Paul Modrich from United States and Aziz Sancar with both American and Turkish nationalities for their contributions in revealing the mechanisms of DNA repair in cells. This article overviewed their research discoveries with the highlighted significance. Meanwhile, as DNA repair is closely connected to cell genome stability, especially under disease conditions, we commented the impact and perspective of high-throughput sequencing and the omic technologies on the future studies in biomedical fields.

A novel therapy against parasites——the Chinese scientist first won the Nobel Prize in Physiology or Medicine 2015

Zhao Limei, Cheng Yuli, Zhu Xinping

2015, 36(5):  829-833.  doi:10.3969/j.issn.1006-7795.2015.05.033

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The 2015 Nobel Prize in Physiology or Medicine is awarded jointly to Tu Youyou, etc three scientists for their discoveries concerning a novel therapy against malaria and infections caused by roundworm parasites. Tu's outstanding contribution is for her discovery of the new anti-malarial drugs—artemisinin and dihydroartemisinin

The analysis of top 10 highly cited papers published in Journal of Capital Medical University(2001-2010)

Zhang Junmin, Wang Xiaomin

2015, 36(5):  834-837.  doi:10.3969/j.issn.1006-7795.2015.05.034

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In order to improving the quality and academic influence of Journal of Capital Medical University,we analyzed the top 10 highly cited papers published in Journal of Capital Medical University(2001-2010),based on the data from CNKI.The results showed that review papers(5 papers) and research papers (4 papers) are among the top 10 highly cited papers, those authors are advanced experts; their hot research were focused on the field of epidemiological investigation in cardio-cerebral vascular disease, Chinese Medicine for the treatment of diseases,evidence-based medicine method and Traditional Chinese Medicine therapeutic evaluation, prevention and control of emerging infectious diseases et al.This study was providing suggestions for topic selecting and column planning.