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Table of Content

    21 June 2010, Volume 31 Issue 3
    消化疾病发病机制和治疗的进展
    The Beneficial Role of Endogenous H2S in Pathogenesis of Nonalcoholic Steatohepatitis Rats
    WU Yan-ning;CAI Zhao-hua;SUN Hai-mei;SHANG Hong-wei;HAO Gang;ZHANG Li-xin;SUN Lin;ZHANG Hua;DING Hui-guo
    2010, 31(3):  287-292. 
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    Objective To investigate the production of H2S and its protective role against mitochondrial dysfunction in rats with nonalcoholic steatohepatitis(NASH). Methods Twenty-six male Spague-Dawlay rats were randomly divided into 3 subgroups: normal group(n=6), NASH model(n=10), and compound fetal bovine liver extract treatment group(n=10). The NASH model was established by feeding rats with high fat diet for 12 weeks. The rate of production of H2S was detected by methylene blue spectrophotometry and CSE mRNA was tested using RT-PCR. The mitochondrial morphology was assessed by transmission electron microscopy and the fragment of mtDNA, ND1, ND6, CO1, CYB and ATP6 were respectively tested by PCR. The hepatic steatosis and inflammation were evaluated using HE and Oil Red staining. Results In NASH rats, the hepatic tissue showed diffuse hepatic steatosis as lipid vacuolar degeneration with inflammatory cell infiltration, and the mitochondria showed swelling, becoming round, markedly abnormal morphology. The rate of liver production of H2S in NASH rats was(1.26±0.08)nmol/min,which was significantly lower than that〔(2.11±0.17) nmol/min〕 in normal group(P<0.05). No significant difference in H2S production between normal and treated group was observed. The decreased hepatic CSE mRNA and its protein were also observed. The level of mtDNA fragment, ND1, ND6, CO1, CYB and ATP6 in NASH rats were decreased. Conclusion The decreased hepatic H2S production and CSE mRNA may be related to mitochondrial injury in NASH rats. Endogenous H2S may play a beneficial role in pathogenesis of NASH.

    Differential Effects of Exogenous and Endogenous Connective Tissue Growth Factor on Rat WB-F344 Cells
    YANG Ai-ting;WANG Ping;LIU Tian-hui;CONG Min;BAI Yan-feng;CONG Rui;WU Peng;YOU Hong
    2010, 31(3):  293-298. 
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    Objective Hepatic progenitor cells(WB-F344) can be differentiated into hepatocytes or cholangiocytes depending on different stimulations. The aim of the current study was to compare influences of exogenous and endogenous connective tissue growth factor(CTGF) on WB-F344 cells differentiation.Methods MTT assay was used to detect cell viability after recombinant human CTGF stimulated WB-F344 cells in complete medium for 24 hours. Western blotting was used to detect CTGF influence on WB-F344 cells differentiation: including the changes of αfetoprotein(AFP), albumin and cytokeratin-19(CK-19). The plasmid expressing CTGF(dl6-95-CTGF) was constructed through inserting the full length of CTGF gene into the AAV vector dl6-95. The stable transfected WB-F344 cells with dl6-95-CTGF using G418 selection. Western blotting was also used to detect the changes of hepatic progenitor markers. Results Twenty-four hours after stimulating WB-F344 by recombinant human CTGF, the viability of WB-F344 was slightly inhibited(P<0.05) without dosedependent manner. High dose(100 ng/mL) of CTGF could change the phenotype markers of WB-F344 cells at protein level: the expression of fetal hepatocyte marker(AFP) was down-regulated, while the expression of mature hepatocyte marker(albumin) and cholangiocyte marker(CK-19) were up-regulated. The plasmid expressing CTGF(dl6-95-CTGF) was constructed and G418 resistant clones were screened for cell clones with stable transfection successfully. Compared with transfected with dl6-95, WB-F344 cells transfected with dl6-95-CTGF could decrease the expressions of AFP and CK-19, while the expression of albumin increased(P<0.05), the results were consistent with CTGF directly stimulated, but the degree of induction was less obvious than that of exogenous CTGF. Conclusion Compared with endogenous CTGF, exogenous CTGF was able to stimulate WB-F344 cells differentiation toward hepatocytes more obviously.
    The Correlation between the TIM-3 Gene Promoter Polymorphisms and the Risk of Gastric Cancer
    ZHU Sheng-tao;CAO Bang-wei;XU Chang-qing;LI Qin;ZHAO Zhi-hai;LI Dan-ping;ZHANG Shu-tian
    2010, 31(3):  299-303. 
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    Objective This study was performed to elucidate whether the polymorphisms within the tumor immunity-regulated gene TIM-3 promoter region were associated with the risk of gastric cancer in the Han ethnic group of China. Methods The association was analyzed in a hospital-based case-control study of 190 gastric cancer cases and 216 cancerfree controls. DNA was obtained from blood samples and the polymorphisms of -574G/T, -882C/T, and -1 516G/T within TIM-3 gene promoter region were genotyped by the method of PCR-RFLP. The correlation between the risk of gastric cancer with the genotype of the polymorphisms was assessed by the odds ratio(OR) values and their 95% confidence intervals(CI). Unconditional Logistic regression model adjusted by the confounding factors was used to analyze the statistical association of the genotypes in two groups by using the SPSS 11.5 software. Results The results of PCR-RFLP showed that the three polymorphisms in TIM-3 gene promoter region were present in our study population. But the homozygous form of the polymorphisms almost did not appear. Statistical analysis revealed that the allele frequency of the three polymorphisms displayed significant difference between the gastric cancer patients and healthy controls(P<0.05). Logistic regression analysis adjusted for confounding factors exhibited that all the three loci genotypes possessed statistically significant difference(P<0.05): loci -574G/T(OR=3.92, 95% CI: 1.40-10.94), loci -882C/T(OR=3.20, 95% CI:1.22-8.41), and loci -1 516G/T(OR=2.32, 95% CI: 1.23-4.40), respectively. Conclusion Although the three polymorphic homozygous genotypes were very rare, the results still revealed that the polymorphisms within TIM-3 gene promoter region were significantly contributed to the genetic susceptibility of gastric cancer in the Han ethnic group of China.
    Adiponectin Modulates MMP-13 and TIMP-1 Expression via p38MAPK Pathway in Rat Hepatic Stellate Cell Line
    CHEN Hui;MA Chao;ZHANG Yu-xian;MA Hong
    2010, 31(3):  304-309. 
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    Objective To investigate the effects of adiponectin on adiponectin receptor 1(AdipoR1) gene and protein expression in hepatic stellate cells(HSCs), and to evaluated the changes of p38MAPK pathway and its influence on matrix metalloproteinase-13(MMP-13) and tissue inhibitor of metalloproteinase-1(TIMP-1) expression in HSCs on the treatment of adiponectin. Methods HSCs proliferation was detected by MTT assay. AdipoR1, MMP-13 and TIMP-1 mRNA levels were detected by real-time RT-PCR. Western blotting was used to detect AdipoR1, MMP-13, TIMP-1, phospho-p38MAPK and phospho-pATF-2 protein expressions in HSCs. Results Adiponectin at concentrations 0.5 μg/mL-2.0 μg/mL had no effect on HSCs proliferation, however, AdipoR1 gene and protein expressions was up-regulated in adiponectin groups compared with control group. Adiponectin could increase phospho-p38MAPK protein expressions in dose and time dependent manners and had a maximal effect after 120 min treatment with adiponectin. Compared with control group, adiponectin could significantly increase MMP-13 gene and protein expressions, moreover, the blocker of p38MAPK could inhibit this effect of adiponectin in HSCs. The mRNA and protein level of TIMP-1 was decreased on the treatment with adiponectin, and this effect of adiponectin could be inhibited by p38MAPK's blocker. Conclusion Adiponectin could up-regulate AdipoR1 expression in HSCs. MMP-13 and TIMP-1 expressions in HSCs could be regulated by adiponectin through activation of p38MAPK pathway.
    Change of Intestinal Motility and Intestinal Mucosal Impairment in Rats with Cerebral Arterial Thrombosis
    CHUO Jia-ran;ZHU Yu-qun;XU Li-xin;YUAN Fang;XU You-qing;WANG Chen
    2010, 31(3):  310-314. 
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    Objective To determine the change of intestinal mucosa, intestinal motility and their relationship. Methods Forty-eight Wistar rats were randomly divided into control group(n=8) and middle cerebral artery occlusion(MCAO) group(n=40). MCAO group was further divided into 3 h, 6 h, 12 h, 24 h, 48 h after the cerebral arterial thrombosis, each subgroup had 8 rats. Along with the application of transluminal techniques of Longa to create the focal cerebral ischemia model in rats. The damages of intestinal mucosa were observed under light and electron microscopy and determine the intestinal motility at each special time point using the methylene blue staining method. Results There was significant difference in the intestinal motility between control and MCAO groups(P<0.05). There was statistical significance of difference between 3 h group and 6 h group(P<0.01), and there was no statistical significance of difference between 6 h group and 12 h group, 12 h group and 24 h group, 24 h group and 48 h group(P>0.05). Damage of intestinal mucosa could be demonstrated via pathological analysis, such as, engorgement, thickening, decurtation, denudation and so on. Conclusion After cerebral arterial thrombosis, damages of intestinal mucosa and intestinal dyskinesis may occur, the above two may be cause and effect for each other.
    Comparison of Changes in Visceral Sensitivity between Mice and Rats after Acute Intestinal Trichinella Infection
    WANG Wen-feng;WANG Huan;QIAN Wei;HOU Xiao-hua
    2010, 31(3):  315-319. 
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    Objective Many studies show that acute intestinal infection plays an important role in the pathogenesis of irritable bowel syndrome (IBS), and many researchers studied the pathogenesis of IBS by creating animal model of visceral hypersensitivity. But few observed the similarities and differences of evolution of visceral sensitivity after infection in different types of animal models.By comparing the similarities and differences of evolution of visceral sensitivity in mice and rats after intestinal infection, this study aimed to clarify that similar factors may lead to visceral hypersensitivity in different ways. Methods Mice and rats were infected with Trichinella spiralis. The weight of the mice and rats at 2 weeks and 8 weeks after infection were measured. Visceral sensitivity of colorectal distension in mice and rats were assessed by abdominal withdrawal reflex(AWR) at different time points before and after infection. Tissues of jejunum, terminal ileum, proximal colon and distal colon were collected after mice and rats were sacrificed. Histopathological changes and inflammation were assessed with HE staining. Results The weight growth rates of mice and rats at 2 weeks after infection were lower than that in the control group(P=0.021, 0.092, respectively). The weight growth rates of mice and rats at 8 weeks after infection had no significant differences as compared with control groups(P=0.596, 0.827, respectively). In both mice and rats, The intestinal pathologic scores in 2 weeks groups were higher than that in control groups; the 8 weeks groups recovered from the intestinal inflammation , and had no significant differences with control groups. At 40, 60 mmHg, in mice , the AWR scores of all the infectious groups were higher than that in the control group(at 40 mmHg, P=0.000, 0.010, respectively; at 60 mmHg, P=0.000, 0.014, respectively), and in 2 weeks group increased more significantly; In rats, the AWR scores of 2 weeks group was lower than that in control group(P=0.046, 0.035, respectively), and the AWR scores of 8 weeks group was much higher than that in control group(P=0.009, 0.000, respectively) and 2 weeks group(P=0.005, 0.001, respectively). Conclusion The intestinal inflammation in mice and rats subsided 8 weeks after Trichinella spiralis infection, visceral hypersensitivity remained; however, the processes of them were different. Therefore, similar intestinal inflammation may lead to visceral hypersensitivity in different ways.
    食管鳞癌的预防及早期诊断和干预
    Progresses in Studies on Endoscopic Diagnosis of Early Esophageal Squamous Cell Carcinoma
    HE Yuan-long;JIAO Yue;ZHANG Shu-tian
    2010, 31(3):  320-323. 
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    Esophageal squamous cell carcinoma(ESCC) is one of the most common malignancies in China. In China, most ESCC patients diagnosed by endoscopy were at advanced stages, which lead to poor prognosis. It is urgent to diagnose ESCC at its early stage. In recent years, many endoscopic methods were developed, such as narrow band imaging(NBI), optical coherence tomography(OCT) and confocal endoscopy. To help endoscopists recognize early ESCC better, we provide a review on the progress of endoscopic diagnosis of early ESCC.
    Effects of Promoter Methylation of Cyclooxygenase-2 on Lower Expressed Esophageal Squamous Cell Line
    WANG Qing-gang;ZHU Sheng-tao;ZHANG Shu-tian
    2010, 31(3):  324-328. 
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    Objective To determine the relationship between cyclooxygenase-2(COX-2) expression and its promoter methylation status in lower expressed COX-2 esophageal squamous cell carcinoma(ESCC), as well as to investigate the significance of combining demethylating agent with selective COX-2 inhibitor in depression of ESCC. Methods ESCC cell line TE-1 was used in this study, and 5-aza-deoxycytidine(50-aza-DC) and nimesulide were added into the cultured TE-1 cell line. Promoter methylation status of cyclooxygenase-2 was examined by bisulfate sequencing analysis. RT-PCR was used to determine the expression of COX-2 mRNA. Cells proliferation of ESCC was tested with MTT reduction assay. Prostaglandin E2(PGE2 ) value was examined by ELISA.
    Results Lower expression of COX-2 mRNA was found in TE-1 cells, while its promoter methylation status was high(58.3%). After treatment with 10 μmol/L 5-Aza-DC, the methylation status was decreased to 30%, on the other hand the mRNA level was increased. Single treatment with nimesulide showed lower suppressing effect compared to combined interference, the reduction value was 16.2% vs 48.63%, respectively(P<0.05). The concentration of PGE2 was raised after 5-Aza-DC treatment from 553.96 pg/mL to 647.54 pg/mL.
    Conclusion For lower expression of COX-2 ESCC, promoter hypermethylation may be a mechanism to control gene expression. Demethylating could increase COX-2 mRNA expression. Combined treatment of ESCC with 5-Aza-DC and selective COX-2 inhibitor may be an alternative choice for depressing ESCC.
    Nuclear Factor-κB Signaling Pathway Constitutively Activated in Tobaccorelated Esophageal Squamous Cell Carcinoma
    ZHOU Qiao-zhi;LU San-jun;WANG Xing;ZHU Sheng-tao;LV Fu-jing;ZHANG Shu-tian
    2010, 31(3):  329-334. 
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    Objective To investigate the influence of cigarette ethanol extraction(EE) and nuclear factor of kappa B(NF-κB) inhibitor, pyrrolidine dithiocarbamate(PDTC), on the constitutive NF-κB activity. Methods EE were incubated with two esophageal squamous cell carcinoma(ESCC) cell lines for 24 h respectively, which were pretreated with or without PDTC, proliferation rate was measured by MTT, apoptosis rate was measured by AnnexinV+PI, NF-κB activity by electrophoretic mobility shift assay(EMSA). Expression of NF-κB and the correlation between smoking were investigated in ESCC specimens from 49 cases by immunohistochemistry. Results PDTC(25~200 μg/ml) inhibited both ESCC cell lines proliferation in a dose-dependent manner. Cell proliferation was increased and cell apoptosis was inhibited by EE in a dose-dependent manner in both ESCC cell lines. EE could induce stimulation on NF-κB activity in both ESCC cell lines. The positive rate of NF-κB expression in specimens of human ESCC was 79.59%. The positive rate of NF-κB expression was higher for smokers compared to nonsmokers. Conclusion EE was shown to be a positive regulator of the activation of the NF-κB pathway in ESCC cell lines. PDTC, a specific inhibitor of NF-κB, can block the effect of EE. NF-κB is activated in ESCC tissue, cigarette smoking may be strongly associated with NF-κB activation.
    Effect of Smoking and Drinking Alcohol on Esophageal Mucosal Low-grade Neoplasia in Wu’an City: A Case-control Study
    LIU Jia;LI Peng;ZHANG Shu-tian;WANG Yong-jun;LV Fu-jing;LI Wei;WANG Qing-gang;ZHAO Hai-ying;LIU Xuan;ZONG Ye;ZHOU Qiao-zhi;JIAO Yue
    2010, 31(3):  335-338. 
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    Objective To investigate the effect of smoking and drinking alcohol on esophageal mucosal low-grade neoplasia in Wu'an city. Methods A 1: 4 age-matched casecontrol study was conducted and questionnaire survey was used to obtain information, then Logistic regression and stratified analysis were applied to calculate odds radio and 95% confidence interval. Results Smoking was the risk factor of esophageal mucosal low-grade neoplasia, OR value was 2.282〔95% CI, 1.164-4.471], P<0.05. Further stratified analysis of smoking group showed that the risk of smoking 1 to 10 cigarettes per day suffering from esophageal mucosal low-grade neoplasia was 3.450 times higher than that of non-smokers, whereas for those who smoked more than 11 cigarettes daily, the risk increased to 4.025 times that of nonsmokers. Smoking and esophageal mucosal low-grade neoplasia was correlated in a dose dependent manner. Drinking alcohol was not the risk factor of esophageal mucosal low-grade neoplasia. Conclusion In Wu'an esophageal cancer high incidence area, smoking was the risk factor of esophageal mucosal low-grade neoplasia, and had positive dose response relationship.
    Endoscopic Screening for Esophageal Cancer in Wu'an City
    LI Peng;ZHANG Shu-tian;WANG Yong-jun;LV Fu-jing;LI Wei;WANG Qing-gang;ZHAO Hai-ying;LIU Xuan;ZONG Ye;ZHOU Qiao-zhi;JIAO Yue;et al.
    2010, 31(3):  339-342. 
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    Objective To establish an esophageal cancer high incidence screening area in Wu'an City, Hebei Province, a pilot study was carried out before largescale endoscopic screening. Methods Home by home survey was launched in Shangtuancheng village, Shangtuancheng Town, Wu'an city, and 40~65 year-old residents were registered to conduct a questionnaire survey. After obtaining informed consent, endoscopy for esophageal cancer and precancerous lesions was performed. Results Among 92 residents who had been surveyed, 88 accepted endoscopy, 19 residents were suspected of having esophageal cancer and precancerous lesions were biopsied, and sent to department of pathology; 7 cases of low or moderate dysplasia, 2 cases of high-grade dysplasia/carcinoma in situ, and 1 case of advanced esophageal cancer were confirmed pathologically.Conclusion This study confirmed the feasibility of esophageal cancer screening in high incidence area, which helps accumulate experience for the future long-term research.
    Preparation of Polyclonal Antiserum against Human PINK1 and its Expression in Rat Brain
    LI Ji-sheng;LI Yao-hua;LI Xin;YU Shun;WANG Nian-qiang;GAO Hua;SUN Xiao-hong;YANG Hui
    2010, 31(3):  343-347. 
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    Objective To investigate expression of Parkinson disease(PD) related protein PINK1 in rat brain. Methods To obtain the antiserum against human PINK1, two New Zealand white rabbits(2.5 kg) were immunized with recombinant Histag fusion human PINK1 protein; the antiserum was purified by salt fractionation, the specificity and sensitivity of the antiserum were verified by Western blotting. The location and distribution of the protein were further determined by immunohistochemistry. Results The human recombinant PINK1 gene was highly expressed as inclusions in prokaryotic expression system. After first salt fractionation, western blotting analysis demonstrated that the antiserum detected a 40 000 band in recombinant human PINK1 protein, a 40 000 band and a 66 000 band in rat brain homogenates which is anticipated. Immunohistochemical staining on normal adult rat brain sections showed that the PINK1immunoreactive substance presents in rat brain bulbus olfactorius, cortex, hippocampus, striatum, cerebellum, substantia nigra and red nucleus. Conclusion A polyclonal rabbit antiserum against human PINK1 was prepared and the immunoreactive PINK1 substance was widely distributed in rat CNS. The antiserum provided a useful experimental tool for profound studies on the tissue expression profile, intercellular location and biological function of PINK1.

    Fluorescent Interaction between Kaempferol and Bovine Serum Albumin
    LI Xiao-rong;WANG Li-juan;LI Yu-hang;CHEN Yi;HU Xiao-min;FAN Yuan-jie;SONG Jian-rong;XUE Ming
    2010, 31(3):  348-352. 
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    Objective To study the characteristics of interaction between the kaempferol(KF) and bovine serum albumin(BSA). Methods The quenching mechanism of the fluorescence of BSA by KF was studied with fluorescence spectra. The dynamic and static quenching constants were determined by the revised Stern-Volmer equation. The number of binding sites was calculated with double logarithmic equation and the main binding force was discussed by thermodynamic equations. The binding distance and energy transfer efficiency between donor(BSA) and acceptor(KF) were obtained based on Forster's nonradiative energy transfer theory. Results KF effectively quenched fluorescence of BSA via a combination of static and dynamic quenching processes. The binding constant KA was calculated to be in the order of 108, indicating a strong interaction between KF and BSA. The number of binding site is approximately equal to 2, the binding distance is 1.45 nm, the energy transfer efficiency is 0.73, and the binding force is mainly hydrophobic force. Conclusion KF effectively quenched the intrinsic fluorescence of BSA via combination of static and dynamic quenching mechanism, and the binding is mainly driven by the hydrophobic interaction.

    Overexpression of BCL10 is Anti-apoptotic to Splenic Marginal Zone B Cells Resulting in Their Hyperplasia
    WU Li-feng;YANG Yi-shu;CHEN Yan;YAN Zhuo-hong;WANG Shuang-li;SUN Min;ZHANG Quan-geng
    2010, 31(3):  353-358. 
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    Objective To explore the molecular mechanisms of MALT lymphoma caused by BCL10 over-expression. Methods BCL10 transgenic (Tg ) mice were identified by BCL10 transgene specific PCR, and BCL10 protein expression were verified by Western Blot. MALT lymphoma precursor cellsmarginal zone(MZ) B cells expansion in BCL10 Tg mice were defined by histology methods and flow cytometry analysis, anti-apoptotic effects of separated MZ B cells were examined with Annexin V-FITC / PI staining. Results The BCL10 Tg/+ mice used in the experiment were verified by transgene specific PCR. Protein expression of transgene is almost at the same level as endogenous BCL10, which means, in Tg/+ mice, BCL10 protein has doubled than that in nontransgenic WT mice. By histology, we found B cells expansion in Tg/+ mice which look like MZ B cells morphologically. By verification with CD21,CD23 staining followed by flow cytometry analysis, it is proven those expanded B cells are CD21high/CD23low MZ B cells. To investigate the mechanisms causing the MZ B cells expansion, we separate the MZ B cells from both Tg/+ and WT control mice by cell sorting with CD43, CD23 double staining. About 30% separated MZ B cells from Tg/+ mice can survive at least up to one week in RPMI-1640 containing 5% FBS, but those from WT mice almost all died at day 3(P<0.01), this result means overexpression of BCL10 in MZ B cells causes anti-apoptotic effects to those cells. We studied pathways of antiapoptotic that BCL10 might be involved by inducing apoptosis with anti-IgM, BCL10 over-expression protect antiIgM caused cell apoptosis. Conclusion Over-expression of BCL10 causes expansion of MZ B cells which have anti-apoptotic property. This property is involved antigen receptor pathway. These results may partially answer the molecular mechanisms of MALT lymphoma with t(1; 14) (p22; q32) chromosome translocation.

    Expression of ACE2 in Mouse Pancreas Tissue
    GUO Li-min;WANG Xia-lian;LIN Shan-shan;Yang Jin-kui
    2010, 31(3):  359-362. 
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    Objective This study was designed to investigate the expression of angiotensin-converting enzyme 2(ACE2) in endocrine and exocrine tissue of mouse pancreas. Methods The expressions of ACE2 in mouse pancreas was detected by reverse transcriptase polymerase chain reaction(RT-PCR) and the expressions of ACE2 in mouse pancreas was detected by immunoelectron microscopy. Results The expression of ACE2 mRNA was detected in whole mouse pancreas and lung tissues, while ACE2 protein was detected in both endocrine tissue and exocrine tissue of mouse pancreas. Conclusion These data demonstrated a tissue specific pattern of ACE2 expression in mouse pancreas and this indicated ACE2 might be present in islet of pancreas of human. Since ACE2 works as a negative regulator of the angiotensin system, the expression of ACE2 protein in endocrine tissue of pancreas may give us more clue for the treatment of diabetes.

    Development of Chinese Speech Test System
    WU Wen-fang;ZHANG Hua;CHEN Jing;CHEN Jian-yong;LI Yu-ling;LIN Chang-yan
    2010, 31(3):  363-367. 
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    Objective To make speech audiometry clinical work, comply with information society development, and to integrate with the international hearing study, the authors developed a computerized speech test system.
    Methods The system used Visual Studio 2008, Access 2003 and DirectX 9. The system included five functions: listener management, speech test, list management, data management and system settings. Results By using listener management, the tester was able to search, review, add, revise and delete listener's information. The speech test included the measurement of a speech recognition score(SRS) and speech recognition threshold(SRT), in an open set test. At the end of a list or series of tests, both the stimuli and corresponding subject responses were recorded onto the hard drive for each test item. In addition, the system could report and print the results in a graphic format. Using list management, testers could review, add, revise and delete lists or items such as characters, key words and sounds. By using data management, the results could be reviewed, searched and exported. ccording to the test condition and the preferences, tester could set system. Conclusion The study changed the traditional manual audiometry mode, might provide a unified platform for speech audiometry in China, and contribute to scientific, standardized, and informationally developed speech audiometry.

    Change of α-Synuclein Level in Rat Brain and Plasma after Ischemia/Reperfusion
    JIA Chun-song;LI Xin;LI Yao-hua;LIU Guang-wei;YIN Juan-juan;WU Yan-chuan;YU Shun
    2010, 31(3):  368-372. 
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    Objective To explore the change of α-synuclein(α-Syn) in rat brain and plasma after ischemia/reperfusion. Methods Thirty adult male Wistar rats were randomly divided into 2 groups: sham operation group(10 cases) and ischemia/reperfusion group(20 cases). A model of middle cerebral artery occlusion(MCAO) was induced by the intraluminal suture for the latter group. After 1.5 h ischemia, reperfused for 24 h, then the level of α-Syn in the plasma and penumbra, infarct core and their controls were measured by Western blotting and ELISA method. Results The level of α-Syn was increased significantly in the penumbra, but did not change obviously in the infarct core or plasma. Conclusion α-Syn was increased in the penumbra of brain after ischemia/reperfusion.

    Superiority of the Multilevel Model in Stratified Sampling Research
    HENG Ming-li;WANG Hong-wu;MA Lin-mao;CHEN Ying-dan
    2010, 31(3):  373-376. 
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    Objective To explore the superiority of the multilevel model compared with the traditional model in crosssectional survey research data by stratified sampling. Methods Using the data of Guangdong in “the Survey of the Second National Important Parasitic Diseases”, the paper analyzing the factors of Clonorchis sinensis disease infection with traditional logistic regression model and multilevel logistic regression model separately. Results The age group is not significant in traditional logistic regression model but becomes significant in multilevel logistic regression model. Conclusion Compared to the traditional logistic regression model,multilevel models can efficiently analyze cross-sectional study data, according to error from different levels while analyzing hierarchical structure data.

    Investigation of Relationship between Suicidal Tendency and Mental Stress in Medical Students
    WANG Jian;KONG fan-zeng;YAN Kai
    2010, 31(3):  377-380. 
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    Objective To study the cause of mental stress developing and its influencing factors in medical school students, give suggestion to them in their health education and prevent from commit suicide. Methods Study population were selected by cluster sampling, information about the students were gathered by questionnaire and analyzed by SPSS 15.0 software package. Results Total detection rate of Suicidal Ideation was 11.62% in study population, and there was no significant difference between boy and girl. The main causes of developing mental stress were Employment Stress, Economy Stress, Social Communication Stress, Social Adaptation Stress and Ability Stress. Among the people characteristic, Single Parent was more important than others. Conclusion When easing the psychological pressure of medical students, priority should be given to employment and economic factors while emphasizing different individual characteristics.

    Expression, Purification and Immunogenicity of the Recombinant Antigen of the Hepatitis A and Hepatitis E Viruses and Comparison with the Traditional Vaccine against HAV
    PANG Yuan-yu;LI Hong-jun;KUI Xiang;XIE Tian-hong;ZHANG Guang-ming;SUN Mao-sheng
    2010, 31(3):  381-387. 
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    Objective To compare the immunogenicity between the genetically engineered recombinant antigen and the traditional vaccine against hepatitis A virus(HAV) and study the ability of expressing different antigens by the same vector and the interaction among the different antigens. Methods Plasmid pBV220-EAAg342 and pBV220-AEAg342 constructed previously which code for HAV vp1 aa24-171 and hepatitis E virus(HEV) ORF2 aa431-615 genes were amplified and identified, then transformed into E.coliBL21 and induced to express the target protein. The expressed proteins were purified by DEAE-Sepharose and CM-Sepharose, identified by SDS-PAGE and Western blotting. KM mice were immunized with the purified recombinant antigen and the traditional HAV inactivated vaccine and the attenuated live vaccine by subcutaneous route and the specific antibody level was determined by ELISA. Results The molecular mass of the expressed recombinant antigen was about 36 000; the expressed product was about 25% in total bacterial proteins. The recombinant EAAg342 could assemble into virus-like particles(VLP) of 10~20 nm radiuses. It indicated that the fusion protein could have specific reaction with HAV and HEV positive sera by Western blotting. The geometric mean titers(GMT) of antiHEV IgG antibody induced by both different recombinant antigens were high. However the levels of anti-HAV IgG antibody induced by the two recombinant antigens were lower than the one induced by the traditional HAV vaccines. Conclusion The recombinant antigen of the hepatitis A and hepatitis E viruses could be expressed in high yield by procaryotic expression system. The two different fusion proteins both have good antigenicity and immunogenicity respectively. The data provided experimental confirmation for development of bivalent vaccine and serologically diagnostic kit for HAV and HEV.

    Molecular Typing of Salmonella Paratyphi A Strains from Guiyang of Guizhou Province with Pulsed Field Gel Electrophoresis
    YAO Guang-hai;WANG Tao;DIAO Bao-Wei;ZHOU Hai-jian;TIAN Ke-cheng;YOU Lv;WEI Xiao-yu;TANG Guang-peng;WANG Ding-ming;KAN Biao
    2010, 31(3):  388-392. 
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    Objective To type and group the Salmonella paratyphi A strains isolated in Guiyang from 2000 to 2007 and to understand the epidemiological features caused by the pathogen. Methods Pulsed field gel electrophoresis(PFGE) of large restriction fragments of bacterial chromosomal DNA was applied to type the Salmonella paratyphi A strains. Results Ten PFGE patterns were identified among the 146 S. paratyphi A isolates, where 32 isolates belonged to pattern 1(21.9%), 73 isolates to pattern 2(50.0%), 13 isolates to pattern 3(8.9%), 7 isolates to pattern 4(4.8%), 14 isolates to pattern 5(9.6%), 3 isolates to pattern 10(2.7%) and each of the remaining 3 isolates to an independent pattern(0.7% respectively). The PFGE type of strains varied year by year, patterns 1 and 2 was domain epidemic patterns; 24 strains from 2 outbreaks had 5 PFGE patterns and the proportion of pattern 1 was 54.2% and the strain isolated from a healthy carrier belonged to pattern 1. Strains isolated from high epidemic areas had 9 PFGE patterns based on the size and number of restriction fragments and patterns which were digested by Spe I. Conclusion The findings of the present study verified the usefulness of PFGE in characterizing and comparing strains of Salmonella Paratyphi A, and forecasting outbreak. The results indicated that although the distribution of PFGE patterns varied among different counties of Guiyang,the pattern 1 and pattern 2 were the epidemic domain, which were responsible for paratyphoid fever in high epidemic counties.

    Prognostic Factors in 337 Inpatients with Liver Cirrhosis
    SUN Li-dong;ZHANG Mei;TANG Zhe
    2010, 31(3):  393-397. 
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    Objective To study the risk factors for poor prognosis in patients with liver cirrhosis. Methods A total of 337 cirrhosis patients were included in the study. The hospital records were analyzed retrospectively, and all patients were followed up by phone call or mail. The associated clinical and laboratory variables were studied by univariate and multivariate statistical analysis. Results ① The survival rates at 3-months, 6-months, 1-year, 3-years and 5-years after hospitalization were 89.0%, 87.8%, 82.2%, 77.4% 59.1% and 46.9%.② Univariate analysis shows that the factors associated with mortality during hospitalization were serum total bilirubin, direct bilirubin, albumin, creatinine, blood urea nitrogen, hemoglobin, prothrombin time, INR for prothrombin time and encephalopathy, hepatorenal syndrome, ascites, and upper gastrointestinal hemorrhage. ③ Univariate analysis showed that the factors associated with mortality 5-years after enrollment were serum total bilirubin, direct bilirubin, albumin, creatinine, blood urea nitrogen, hemoglobin, prothrombin time, INR for prothrombin time, cholesterol, low density lipoprotein, serum sodium and encephalopathy, hepatorenal syndrome, ascites and age. ④ Multivariate analysis showed that serum albumin, creatinine, blood urea nitrogen, total bilirubin, prothrombin time, low density lipoprotein, ascites and age were independently related to 5year mortality. Conclusion ① For cirrhotic patients, a number of clinical factors were significantly correlated with mortality in hospital or 5 years after enrollment. The liver function is the main factor for prognosis. ② Ascites and age were also significantly related to 5year mortality.

    Open Tension free Repair of Inguinal Hernia Analysis of 1 136 Cases
    LIANG Cun-he;WANG Yu;TAN Si-wei;CHEN Zhi-yu
    2010, 31(3):  398-401. 
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    Objective To evaluate the effects of three operation methods on the open tension-free repair of inguinal hernia. Methods The data of 1 136 cases of inguinal hernias seen between Feb. 2003 and Aug. 2009 after tensionfree hernioplasty were analyzed retrospectively. Total expenses, length of hospital stay, post operation complications, rate of recurrence were compared among the groups. Results In all the 1 136 cases the tension-free hernia repair was successful. The operation methods included Lichtenstein, Rutkow, preperitoneal inguinal herniorrhaphy(PIH). The average hospital expense was significantly lower in the Lichtenstein group than in the Rutkow and PIH groups. There were no significant differences in recurrence rate, post operative complications, length of hospital stay among the three groups. Conclusion All the 3 hernioplasty procedures showed ideal effects, characterized by low recurrence rate, minor postoperative pain, quick recovery, short hospital stay and few postoperative complications. Individualized treatment programs are still the optimal choices. Severe early postoperative pain reliably predicted the likelihood of persisting chronic groin pain.

    Clinical Evaluation of the Use of Expandable Intramedullary Nail in Treatment of Peritrochanteric Fracture
    QIANG Hua;LU Meng;ZHAO Er-hong
    2010, 31(3):  402-404. 
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    Objective To investigate and assess the effectiveness and methods of using expandable intramedullary nail system(EINS) in the clinical treatment of femoral intertrochanteric fractures. Methods From June, 2006 to December, 2009, a total of 47 cases with femoral intertrochanteric fracture were treated with EINSFixation PF. In total, 47 nails were used in 20 male patients and 27 female patients at the ages of 68 to 91 years; including 18 cases with stable fracture(A1.1~1.3), 29 cases with unstable fracture(A2.1~A3.3). Results In all 47 cases, the wounds healed within the first stage; time of surgery ranged from 45 to 110 minutes, averaging 68 minutes per surgery. Blood loss ranged from 70 to 300 mL, on average 136 mL. All 47 patients were followed up post surgery. Followup period was from 8 to 26 weeks, averaging 14 weeks. The average healing time was 10 weeks. Conclusion EINS has a unique design that is advantageous in the clinical treatment of femoral intertrochanteric fractures in older people. Its surgical procedures are relatively simple, less invasive, and the time needed for the surgery was shortened. In addition, EINS significantly decreases the amount of X-ray exposure, thus, decreasing the potentially harmful effects of X-ray on both doctors and patients. The expandable stud extrudes cancellous bone and does not pose the problem of bone cutting used in internal bone fixation methods, and leads to a faster fracture healing. EINS is deserved to be widely used in clinical treatment.

    Expression of Tumor Suppressor Gene PTEN in Endometrium of Postmenoposal Women with Breast Cancer Treated with Tamoxifen
    XING Feng-ling;CHEN Xue-qin;FENG Li-min;ZHANG Hua
    2010, 31(3):  405-408. 
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    Objective To investigate the influence of tamoxifen on the postmenopausal women, and test a hypothesis whether tamoxifen can cause endometrial cancer in the postmenopausal women. Methods Using immunohistochemistry(LSAB) on formalinfixed paraffinembedded endometrium sections to study the phosphatase and tension homology deleted on chromosome ten(PTEN) in endometrium of postmenopausal patients with breast carcinoma who took tamoxifen 40 mg/day for over 2 years and in endometral adenocarcioma, with prolapse uterine endometrium as normal controls. Results There was significant difference among Group tamoxifen, Group carcinoma of endometrium and Group normal endometrium(P<0.05). The differences between the groups of tamoxifen and normal endometrium were significant, the same was seen between carcinoma of endometrium and normal endometrium groups. But no significant difference was found between tamoxifen and carcinoma of endometrium groups. Conclusion The PTEN expression of the breast cancer patients who took tamoxifen 40 mg/day for over 2 years was lower than that in the normal people, and was almost the same as that in the patients of carcinoma of endometrium. Because tamoxifen stimulated postmenopause endometrium in the proliferative phase for a long time, thus made the expression of the PTEN decreased. Loss of PTEN expression is an early event in carcinoma of endometrium. Detection of PTEN protein may be a diagnostic biomarker for the earliest endometrial precancers and carcinoma.

    Retrospective Analysis of Treatments of Pregnant Women Complicated with Rheumatic Heart Diseases
    WANG Yi-xin;DING Shu-fang;LIU Hai-hang;WU Jie;LIU Qing;FANG Zhen;ZHANG Jun;LIU Tao
    2010, 31(3):  409-411. 
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    Objective To investigate the effects of different clinical therapies of pregnant women complicated with rheumatic heart diseases. Methods Clinical data of pregnant women complicated with rheumatic heart diseases collected from Beijing Anzhen Hospital from January 2002 to January 2009, were analyzed retrospectively. Results The prevalence of pregnancy complicated with rheumatic heart diseases was 3.91%. One neonatal death occurred in 14 cases who received drug treatment. Of 11 cases treated with heart surgery, 5 cases received valve replacement surgery during progestation, 6 cases received percutaneous balloon mitral valvuloplasty during gestational period. Conclusion The pregnancy complicated with rheumatic heart diseases does seriously harm the maternalfetal health. The heart surgery was effective in treatment of pregnancy complicated with rheumatic heart diseases. Valve replacement surgery during progestation and percutaneous balloon mitral valvuloplasty during gestational period did not show significant difference in improving motherfetal prognosis.

    Analysis on Rules of Traditional Chinese Medicinal Dosage in the
    LI Ye
    2010, 31(3):  412-415. 
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    Objective According to the theory of number and phenomenon, to explore the relationship between dosage and efficacy in “Shang Han Lun”. Methods Through adopting the original text of “Shang Han Lun” and the later generations' analysis of efficacy for each medicine in the different formula, the relationship between quantity and effect was studied by using Logistic regression, paired chisquare test and other statistical methods. Results There is an association between the number of dose and the eight therapeutic methods, but number could not explain the whole effect. Conclusion This study developed a method for studying the laws of applying the traditional Chinese medicines in “Shang Han Lun”, and provides a guideline for further study.

    Analysis of Clinical Characteristics in 20 Patients with AL Amyloidosis
    LIU Ding;LIU Gang
    2010, 31(3):  416-419. 
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    Objective To realize the clinical characteristics of multisystem involvement in patients with AL amyloidosis. Methods The clinical data of 20 patients who were diagnosed as having renal AL amyloidosis, by immunohistochemical method on renal biopsy sections were retrospectively analyzed. Results Of the 20 patients, 90% of were over 40 years old. The ratio of male to female was 3:2. Two or more systems involvement was found in 90% of patients and 30% had more then three systems involved. In this group kidney involvement was found in 100%(nephrotic syndrome 90%), hypotension in 45%, thickened ventricular walls in 35%, abnormal electrocardiogram in 80%, hepatomegaly in 35%, glossal hypertrophy in 25%, body weight loss in 40%, monoclonal light chain in serum or urine in 85% of patients. Conclusion AL amyloidosis often involves multiple systems, including digestive system, kidney and heart were the common target organs.The clinical characteristics of the disease are complicated.

    Surgical Treatment of Cervical Spinal Cord Compressive Disorders in 65 Cases
    CAO Guo-dong;LIU bo;HOU Guan-hua
    2010, 31(3):  420-423. 
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    Objective To explore surgical treatment of the cervical spinal cord compression. Methods From August 2003 to November 2008, 30 cases of cervical spinal cord compression were treated in Beijing Yanqing Hospital and from December 2008 to September 2009, 35 cases of cervical spinal cord compression were treated in Beijing Jishuitan Hospital with different kind of surgical treatment. All the patients were followed up. Results Thirty cases received anterior decompression, another 30 cases received posterior decompression and 5 cases received anterior and posterior approach stage 1 combined surgery. The 65 patients were followed up for 9 months to 5 years(average 3.3 years) in outpatient department or by telephone. According to the JOA, all the patients had satisfactory outcome. Conclusion Posterior spinal canal expansion angioplasty is suitable for developmental cervical canal stenosis, ossification of posterior longitudinal ligament(OPLL) and degenerative spinal stenosis. Anterior cervical decompression and bone graft fusion with titanium plate fixation or cervical artificial disc replacement are suitable for one-two-disc hernia. The anterior approach and posterior approach is selected for spinal stenosis combined with spine cord compressed from front and rear side.