1.Effect of erythropoietin on the proliferation and apoptosis of neonatal porcine islet cells
Honghui HE ; Tianhui WU ; Jing XIONG ; Ke CHEN ; Zhaohui MO
Journal of Central South University(Medical Sciences) 2010;35(11):1115-1122
Objective To investigate the effect of erythropoietin on the proliferation,differentiation,and apoptosis of the cultured neonatal porcine islet cells in vitro.Methods Neonatal porcine islet cells were separated and pured from neonatal pigs with collagenase digestion and tissue culture,and their viability and purity were tested. The neonatal porcine islet cells were divided into a control group and an experimental group.The experimental group was treated with erythropoietin but not the control group,and the insulin secretion responsiveness induced by low and high glucose stimulation in the islet was tested after 5 days. Cells were counted and the activation of amplification was determined by MTT chromatometry. The rates of cell apoptosis were observed by ethidium bromide/acridine orange (EB/AO) of fluorescent light staining and flow cytometry,and the cell cycle was analyzed by flow cytometry. The expression of bcl-2,bax,caspase-3,glucose transporter 2 (GlUT-2),and pancreatic duodenal homeobox-1 (PDX-1) mRNA was tested by RT-PCR.Results After erythropoietin was treated in the cell culture,the neonatal porcine islet cells had normal morphology,function,and reaction of insulin secretion to the glucose stimulation. Cell count showed more cells in the experimental group than in the control group (P<0.05). MTT chromatometry showed the optical absorbance tended to increase with time,and compared with the control group,the optical absorbance was higher in the experimental group (P<0.05),the expression of PDX-1 mRNA was slightly up-regulated (P<0.05). The expression of GLUT-2 mRNA had no difference in the 2 groups (P=0.34). In the experimental group,the apoptisis rate was lower than that in the control group by flow cytometry and EB/AO fluoscence staining (P<0.01),and the expression of bcl-2 mRNA was higher. Howerer bax mRNA and caspase-3 mRNA were obviously lower than those in the control group (P<0.01).Conclusion Erythropoietin can promote the proliferation but has no effect on the function of neonatal porcine islet cells in vitro. Erythropoietin can protect neonatal porcine islet cells from apoptosis through up-regulating bcl-2 mRNA and downreguling bax and caspase-3 mRNA.
2.Multilevel finite element analysis on the biological tribology damage of water on bone tissue
Zedong HE ; Jing ZHAO ; Liangyu CHEN ; Ke LI ; Jie WENG
Chinese Journal of Tissue Engineering Research 2017;21(7):1041-1045
BACKGROUND:Many studies reported the relationship of the mechanical properties and water content about bone tissue, which is one of organizations containing the lowest water content on human body. Researches on effect of water on biological tribology behavior of bone tissue have been rarely reported and are the experimental study generally. OBJECTIVE:To explore the influence and the damage mechanism of water on biological tribology behavior of bone tissue, by comparing multiscale numerical model established with the experiment. METHODS:Dehydration of the bone tissue was studied by nanoindentation test and both reciprocating sliding and impact wear tests. A multi-scale finite element model was constructed under a flat-on-ball configuration. RESULTS AND CONCLUSION:The viscoelasticity and the tribological properties of bone tissue significantly decreased as well as the different wear mechanisms under applied loading after drying. The analytical results indicated that there were high stress condition, which incurred the micro-crack initiation and the appearance of peeling and wear, around the Haversian canal, circumferential lamellas and the interstitial tissues. Meso-scale:dehydration weakened the function of absorption and interruption of stress, which facilitated crack extension in pore. Micro-scale:the high stress gradient of structure of canaliculi and lacunae is an important cause of tissue damage.
3.Transfer of regulatory T cells induced by TGF-β1prolonged the skin-graft survival in mice
Jin YUAN ; Ke WU ; Wentao HE ; Feng WANG ; Zhonghua CHEN
Chinese Journal of Microbiology and Immunology 2008;28(8):680-684
Objective To study the probability of transferring the regulatory T cells induced by TGF-β1 to prolong the allograft survival and the mechanisms involved.Methods According to the different culture conditions.three experimental groups were established:control group(T cells from C57 BL/6 mice cultured with II-2),MLR group(T cells from C57BL/6 mice activated by alloantigen)and TGF-βgroup(T cells from C57BL/6 mice activated by alloantigen and cultured with 5.0 ng/ml TGF-β1).After the culture,the ratio of CI4+CD25+T and the Foxq3 expression were measured by FACS and RT-PCR,respectively.On 9th day,the pathologic analysis was performed and the ratios of TH1,TH2 and Treg and the proliferation of lymphocytes were measured.Results The ratio of CD4+CD25+T in TGF-β group was higher than that in control group and MLR group(P<0.05),and Foxp3 was expressed in CD4+CD25+T cell from TGF-βgroup.After transferring ofthe cells,the allografi survival time in TGF-β group was prolonged and its mean survival time(MST)was(22.8±1.9)d,which was longer than that in MLR group and control group (P
4.Effect of GLP-1 on high glucose-induced human umbilical vein endothelial cell apoptosis and mechanism
Xiaoyan YUAN ; Ke CHEN ; Honghui HE ; Lilin ZHAO ; Zhaohui MO
Journal of Central South University(Medical Sciences) 2013;38(10):1029-1034
Objective: To investigate the effects of glucagon-like peptide-1(GLP-1)on high glucose-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the mechanism involved. Methods: HUVECs were cultured under varying conditions for 48 h, and the cell viability was spectrophotometrically measured by MTT assay. Flow cytometry detected the ratio of cell apoptosis. Western blot detected the protein levels of p-Akt and p-eNOS, while NO assay kit detected the NO concentration.
Results: Treatment of high glucose (33 mmol/L) for 48 h signiifcantly decreased the HUVECs viability and induced the apoptosis of HUVECs, concomitant with decreased Akt and eNOS phosphorylation leves and subsequent NO production. Treatment with GLP-1 (3 nmol/L) for 48 h in the high glucose group increased the HUVECs viability (P<0.01), decreased the ratio of HUVECs early apoptosis (P<0.05), ameliorated the reduced protein levels of p-Akt and p-eNOS caused by high glucose, and increased the NO production (P<0.05). The anti-apoptotic effect and the increased NO production of GLP-1were inhibited by PI3K inhibitor wortmannine (100 nmol/L) or eNOS inhibitor L-NAME (100μmol/L). The effect on p-Akt, p-eNOS of GLP-1 was inhibited by wortmannine (100 nmol/L) while L-NAME (100μmol/L) did not have any influence on the expression of p-Akt.
Conclusion: GLP-1 can ameliorate high glucose-induced HUVECs apoptosis, which is probably related to the up-regulation of PI3K/Akt/eNOS pathway.
5.Clinico-pathologic analysis of 12 cases of epithelioid hemangioendothelioma
Ao XU ; Ke CHEN ; Qi WANG ; Xiaodong HE
Chinese Journal of Clinical and Experimental Pathology 2017;33(3):263-267
Purpose To study the clinicopathologic features,immunophenotypes,diagnosis and differential diagnosis of epithelioid hemangioendothelioma (EHE).Methods Histopathology,clinical features and immunophenotypes of 12 cases of EHE which diagnosed by resection and core needle biopsy were analyzed retrospectively.These patients were followed-up.Results There were 9 males and 3 females,aged from 30 to 69 years,with an average age of 43.5 years.Among 12 EHEs,there were 8 cases in liver,3 cases in bone,and 1 case in lung.Histopathologically,the tumor cells were round and oval epithelioid arranged in single,nests,and cords.Tumor cells had abundant eosinophilic cytoplasm that contained vacuoles and erythrocyte in the tumor cytoplasm.The stroma showed myxoid change.Sparse calcification could be seen in some EHEs.Immunophenotype,tumor cells showed strong positive for CD34,CD31 and factor Ⅷ related antigen (FⅧRAg).Most EHEs are alive with tumor.1 case recurred in 6 months after surgery.1 patient died at 100 months postoperation.3 cases were lost follow-up.Conclusion EHE is a very rare low-grade malignant vascular neoplasm especially for arising from solid organ.Precise diagnosis must depend on surgical histopathology and molecular biology test of WWTR1-CAMTA1 fusion gene.
6.Geldanamycin inhibits proliferation and motility of Her2/neu-overexpressing SK-Br3 breast cancer cells
Yu REN ; Ke WANG ; Jianjun HE ; Wuke CHEN ; Qingyong MA
Journal of Pharmaceutical Analysis 2008;20(4):235-241
Objective Benzoquinone ansamycin antibiotic, geldanamycin (GA), is a new anticancer agent that could inhibit Hsp90 by occupying its NH2-terminal ATP-binding site. This study was to investigate the antitumor efficacy of GA on Her2/neu tyrosine kinase overexpressing human breast cancer cell line SKBr3. Methods The degradation of Her2/neu tyrosine kinase was analyzed by Western blotting, the proliferation index was determined by MTT assay,cell cycle distribution was detected by flow cytometry, Cyclin D1 mRNA transcription was measured by RT-PCR and real-time PCR, and cell motility was evaluated by the cell culture insert model. Results GA induced a dose- and a time-dependent degradation of the Her2/neu tyrosine kinase protein and concurrently, the inhibition of cancer cell proliferation. The antitumor effects mediated by GA included: GA treatment decreased the survival rates of cancer cells,and led to a dase-dependent G1 arrest. Furthermore, this antitumor effect was proved to be related to declined transcription of Cyclin D1. Concurrently, the motility of cancer cells was reduced by GA. Conclusion GA treatment could induce the degradation of Her2/neu tyrnsine kinase efficiently, inhibit cancer cell proliferation and reduce motility in Her2/nen tyrosine kinase overexpressed human breast cancer cell line SKBr3.
7.Etiological characteristics of Streptococcus pyogenes isolated from children with Scarlet fever in Guangdong province, China
Bixia KE ; Baisheng LI ; Hailing TAN ; Changwen KE ; Dongmei HE ; Jingdiao CHEN ; Meizhen LIU
Chinese Journal of Microbiology and Immunology 2013;(5):360-363
Objective To investigate the etiological characteristics of Streptococcus pyogenes that caused scarlet fever in different periods in Guangdong province.Methods 22 isolates from different periods were analyzed through emm typing,PCR detection for super antigen genes,antibiotic susceptibility test and pulsed-field gel electrophoresis (PFGE).Results All isolates were susceptible to cefotaxim,levofloxacin and penicillin.Streptococcus pyogenes isolated after the year 2000 were 100% resistant to erythrocin and clindamycin,but the resistant rate for strains isolated before the year 2000 was 9.1% (1/11).There were 3 emm types indentified from 22 isolates including emm12.0 (59.09%,13/22),emm6.0 (36.36%,8/22) and emm1.0 (4.55%,1/22),which were detected in the isolates from the year 1997 and 2011,from 1978 and 1986,and from 2008,respectively.The positive rates for speA,speB,speC,speF,speG,speH,smeZ,and ssa genes detected by PCR were 54.55%,100%,100%,100%,100%,54.55%,0%,and 86.36% respectively.Among all strains,95.45% of the isolates carried 6 superantigen genes simultaneously.Three clusters of 10 PFGE subtypes were identified in 22 isolates.Cluster Ⅰ consisted of all strains from 1997 and one strain from 2011.Cluster Ⅱ consisted of strains isolated from 1978 and 1986.Cluster Ⅲ consisted of nine strains from 2011 and one from 2008.Conclusion S.pyrogenes isolates in Guangdong province were susceptible to penicillin but resistant to erythrocin.emm 12.0 accounted for the majority of the three types and there was a high frequency of super antigen genes.
8.Antimicrobial resistance patterning and pulsed field gel electrophoresis (PFGE) typing for non -typhoidal Salmonella isolated from diarrhea cases in Guangdong province, China
Baisheng LI ; Bixia KE ; Dongmei HE ; Hailing TAN ; Chen WANG ; Zhaoming LIANG ; Meizhen LIU ; Jingdiao CHEN ; Changwen KE
Chinese Journal of Microbiology and Immunology 2012;32(6):542-548
Objective To investigate the antimicrobial resistance pattern of non-typhoidal Salmonella isolated from diarrhea cases in Guangdong province,China.The multidrug-resistant strains were analyzed by pulsed field gel electrophoresis(PFGE) typing.Methods All the non-typhoidal Salmonella strains isolated between 2009 and 2011 were serotyped,then the antimicrobial resistance was detected by the disk diffusion method and molecular typed by PFGE.Results 91.76% (256/279) S.typhimurium isolates were multiple resistant to 3 and more antimicrobials.Forty S.typhimurium isolates were multiple resistant to 9 and more antimicrobials and 3 out of which were multiple resistant to all the 12 antimicrobials in vitro.96.91% (94/97) Salmonella I4,5,12:i:-isolates were multiple resistant to 3 and more antimicrobials.Nine Salmonella I4,5,12:i:- isolates were multiple resistant to 9 and more antimicrobials and I out of which was multiple resistant to all the 12 antimicrobials1 in vitro.47% (47/100) S.enteritidis isolates were multiple resistant to 3 and more antimicrobials.Only 1 S.enteritidis isolates was multiple resistant to 9 and more antimicrobials.4.27% (27/632) non-typhoidal Salmonella isolates was resistant to ciprofloxacin,including 17 S.typhimurium and 6 Salmonella 14,5,12:i:- isolates.Also,there were 3 1.96% ( 202/632 ) non-typhoidal Salmonella isolates was intermediary to ciprofloxacin.The PFGE patterns of the predominant strains which were highly resistant and multidrug-resistant had different genotypes and demonstrated significant genetic diversity.Conclusion The situation about the multiple antimicrobial resistances of non-typhoidal Salmonella in Guangdong province has showed the prevalent problem.The PFGE types of the multiple drug-resistant strains prompted these strains were come from different clones.This requires that we continue to strengthen the resistance monitoring and control of the rational use of antibiotics.
9.Clinical effect of oxaliplatin-containing and cisplatin -containing regimen in TACE of advanced hepatocellular carcinoma
Yingqiang JIANG ; Yancai CHEN ; Zongwan CHEN ; Lianxi ZHENG ; Ke YANG ; Lauqiong GOU ; Xu LIN ; Ping HE
Cancer Research and Clinic 2009;21(10):687-689
Objective To explore the clinical effect and side-effect of oxaliplatin(L-OHP)-containing regimen and cisplatin (DDP)-containing regimen in TACE of advanced hepatocellular carcinoma (HCC). Methods 108 patients with advanced HCC were randomly divided into experimental group(n=55) and control group (n =53). The experimental group were treated with TACE using L-OHP. After dilute with glucose solution, L-OHP(130 mg/m2) and FT207 (500-750 mg/m2) were injected into blood vessel respectively. ADM (40 mg/m2) and LP (10~30 ml) were emulsified and then used for vessel embolism according to the size of focus. The control group received TACE with DDP, DDP (40 mg/m2) and F]'207(500~750 mg,/m2) were diluted with glucose solution, and also according to the size of tumor' s focus, ADM(40 mg/m2) and LP(10~30 ml) were emulsified for vessel embolism, and then diuretic. Results The total effective rate of experimental group was 67.3 % (37/55), and that of control group was 47.2 % (25/53), and the difference was with statistical significance (P <0.05). The descent rate of AFP of experimental group was 73.1%(31/43), and that of control group was 44.7 % (17/38), and the difference was with statistical significance (P <0.05). The main side effects were gastrointestinal reactions, the incidence rate of nausea and vomiting in experimental group were lower than control group, and the difference was with statistical significance. The incidence rate of leucopenia, damage of hepatic function and peripheral neuritis were not significant. Damage of heart and kidney were not found in the two sets. Conclusion L-OHP -containing regimen in TACE of advanced HCC is an efficient method, with good security and good tolerance to patients.
10.Cross-sectional survey on healthcare-associated infection in a hospital in 2014
Huan LIU ; Ke CHEN ; Qin ZHANG ; Liangbao LIU ; Hongyan HE ; Jiali CHEN
Chinese Journal of Infection Control 2015;(7):483-485
Objective To investigate the current status of healthcare-associated infection(HAI)and antimicrobial usage,so as to provide a scientific basis for improving the management of HAI. Methods A cross-sectional survey was conducted by combination of bedside visiting and medical records reviewing,HAI were investigated among all hospitalized patients between 0:00 and 24:00 on August 21 ,2014.Results A total of 2 216 patients were investiga-ted,the prevalence rate of HAI was 4.83% ,the case infection rate was 5.14% ;the main infection site was lower respiratory tract(63.16% ),antimicrobial usage rate was 39.71% ,the proportion of prophylactic and therapeutic use of antimicrobial agents was 32.27% and 61 .71% respectively.596 patients received therapeutic antimicrobial use,specimen detection rate was 56.21% (n= 335),the detection rate of pathogens was 15.52% (n= 52). The ma-jor detected bacteria were Pseudomonasaeruginosa,Klebsiellapneumoniae,Acinetobacterbaumannii,Escherichia coli,and Stenotrophomonasmaltophilia.Conclusion HAI prevalence survey is helpful for realizing the occurrence of HAI,respiratory tract is the main infection site,gram-negative bacteria is the major pathogen,management of prophylactic use of antimicrobial agents is the focus of HAI management.