1.Stable surface expression of a gene for Helicobacter pylori toxic porin protein with pBAD expression system.
Zhixiang, PENG ; Xi, WEI ; Zhengmei, LIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):435-8
Helicobacter pylori (H. pylori) infection causes peptic and duodenal diseases in humans. Among a 32-protein family of outer membrane proteins, a porin-like protein, HopE, has been a subject of note, mainly for its conservative nature among H. pylori, and for its potential as a vaccine candidate. To achieve stable surface expression of this host cell-toxic protein, hopE gene was introduced into pBAD expression system. After induction with arabinose, all 15 randomly-chosen E. coli LMG 194 colonies from 3 successive passages could express HopE protein, while only 1 from 5 E. coli colonies that contained lac operon-regulated plasmid encoding hopE gene could express HopE. Indirect immunofluorescence confirmed the expression of HopE on E. coli cell surface.
2.Roles of p53 in the interaction with digestive malignant tumors
Peng WANG ; Zheng WANG ; Zhixiang ZHOU
Practical Oncology Journal 2016;30(5):468-472
p53,the widely studied tumor suppressor gene ,plays a key role in regulating cell cycle ,repai-ring DNA damages , eliminating free radicals and regulating immune responses and so on .The recent studies showed that p53 mutation was in close biological connections with the occurrence , development , infiltration and metastasis in over 50 percent digestive systemic malignant tumors .In view of the current researches ,we review the biological features of p53,the relationship between p53 and digestive systemic malignant tumors ,and gene therapy of p53 to provide references for the pathogenesis ,diagnosis,treatment and prognosis of digestive systemic malig-nant tumors in this paper .
3.Analysis of 56 cases of ADR Induced by Irinotecan in Advanced Malignant Cancer Patients of Our Hospital
Peng LI ; Ping SHI ; Zhen WANG ; Zhixiang YANG
China Pharmacy 2016;27(5):635-637
OBJECTIVE:To explore the characteristics and regularities of the adverse drug reactions(ADRs)induced by irino-tecan,in order to provide reference for clinical application. METHODS:In retrospective study,56 cases of irinotecan-induced ADR collected from our hospital during Jun. 2011-Jan.2015 were analyzed statistically in respects of gender,age,disease types, ECOG score,medication,organs/systems involved in ADR,clinical manifestations,outcome,etc. RESULTS:All 56 patients suf-fered from ADR,among which the percentage of hematopoietic system accounted for the highest(100%),and followed by diges-tive system;the grade Ⅰ-Ⅱ ADRs accounted for 87.20%,the grade Ⅲ-Ⅳ ADRs accounted for 12.80%. 64.50% of ADRs oc-curred within 1 week. In respect of conversion,85.71%(48/56)of the patients recovered with supportive care and continued che-motherapy,the remaining 8 cases had to reduce the dose of chemotherapy and then recovered. CONCLUSIONS:With regular dose,irinotecan is well tolerated. Since ADRs induced by irinotecan are slight,all the ADRs are well managed. It is important to enhance follow-up observations of patients to prevent some serious ADRs.
5.Application of comprehensive teaching mode in cataract clinical teaching
Zhixiang DING ; Yanyi PENG ; Wenbin ZHANG ; Yuming ZHANG
Chinese Journal of Medical Education Research 2013;(5):490-493
Objective To explore teaching effect of comprehensive teaching mode of PBL combined with CBL in ophthalmologic cataract clinical teaching.Methods 2008 grade clinical majors (n =80) in class 1 as experimental group were taught by comprehensive teaching mode of PBL combined with CBL while those (n =83) in class 2 as control group were taught by traditional LBL teaching mode.After the courses,the teaching effects of two methods were compared.SPSS 11.5 software was used for statistical analysis,x2 test for satisfaction survey and t-test for theoretical examination scores.The test level is α =0.05.Results There were significant differences between experimental group and control group in improving comprehensive quality and developing clinical thinking.Scores of understanding knowledge,case analysis and total scores of experimental group were higher than those of control group (all P < 0.05).Conclusions Comprehensive teaching mode may improve the teaching effect of cataract clinical teaching,but it need to be explored and improved continually in practice.
6.Up-regulation of DNA-PKcs and Its Mechanism in Human Glioma
Zhixiang ZHUANG ; Liqin SHEN ; Shuyu ZHANG ; Peng QIU
Chinese Journal of Clinical Oncology 2010;37(4):216-219
Objective: To detect the gene expression of Ku70, Ku80, ERCC4, lig4 and DNA-PKcs in non-homologous end joining pathway in human pdmary glioma tissues and normal brain tissues and to explore the underlying mechanism. Methods: The expression levels of Ku70, Ku80, ERCC4, lig4 and DNA-PKcsin in 36 glioma samples and 12 normal brain tissue samples were measured by SYBR Green real-time quantitative PCR. Methylation of DNA-PKcs was detected by methylation-specific PCR (MSP). Results: There was no significant difference in Ku70, Ku80, ERCC4 and lig4 expression between human primary glioma and normal brain tissues (P<0.05), while DNA-PKcs was significantly up-regulated (P= 0.002). The expression of DNA-PKcs was significantly higher in grade Ⅲ and Ⅳ glioma than that in grade Ⅱ glioma and normal brain tissues (P<0.05). Moreover, glioma tissues showed weaker methylation than normal brain tissues. Conclusion: The up-regulation of DNA-PKcs may be associated with pathogenesis of glioma. Demethylation of DNA-PKcs promoter is an important reason for its up-regulated expression in glioma.
7.Risk factors for lower extremity lymphedema following treatment of gynecologic cancers:a meta-analysis
Xin CHANG ; Jiaofeng SHEN ; Qiliang PENG ; Zhixiang ZHUANG ; Ye TIAN
Chinese Journal of Radiation Oncology 2017;26(9):1038-1044
Objective To investigate the multiple risk factors for lower extremity lymphedema in patients following treatment of common gynecologic cancers by meta-analysis for systematic analysis and comprehensive quantitative study.Methods Clinical trials published up until August 2016 were retrieved from PubMed, Embase, and the Cochrane Library.The quality of the included studies was assessed by the Newcastle-Ottawa Scale, and data analysis was performed using Stata 14.0 and RevMan 5.3.The strength of the associations between risk factors and gynecologic cancer-related lower extremity lymphedema was described as odds ratio (OR) and 95% confidence intervals (CI).Results Eighteen studies were included in the meta-analysis, and 8 relevant factors were identified.The risk factors for lower extremity lymphedema after treatment of gynecologic cancer mainly included radiotherapy (OR=2.45, 95%CI:2.05-2.95, P=0.000), FIGO stage (OR=2.29, 95%CI:1.66-3.14, P=0.000), and pelvic lymph node dissection (OR=2.00, 95%CI:1.02-3.91, P=0.040).Conclusions Radiotherapy, FIGO stage, and pelvic lymph node dissection are the main risk factors for lower extremity lymphedema after treatment of gynecologic cancers.
8.Identification of microRNA-217 targeted gene ANLN in pancreatic cancer PANC1 cells
Yin JIANG ; Baohua HOU ; Zhixiang JIAN ; Huiling WANG ; Peng CUI ; Jinrui OU
Chinese Journal of Pancreatology 2013;(3):175-178
Objective To identify the miR-217 targeted gene ANLN by experiment.Methods Bioinformatic algorithms were used to predict the potential targets of miR-217.Then,ANLN binding with miR217 and mutant ANLN (mutANLN) sequence were designed and synthesized,and their amplified fragments were inserted into plasmid psiCHECK-2,and recombinant plasmid psiCHECK-2-ANLN and psiCHECK-2-mutANLN were reconstructed.The two recombinant plasmids were co-transfected into pancreatic cancer cell line PANC1 with miR-217,miR-217 inhibitor,NC,NC inhibitor by liposome,respectively.Dual luciferase reporter system was used to determine the luciferase activity,and Western blot was used to measure the expression of ANLN protein.Results The luciferase activities of psiCHECK-2-ANLN,psiCHECK-2-ANLN +miR-217,psiCHECK-2ANLN + miR-217 inhibitor,psiCHECK-2ANLN + NC,psiCHECK-2-ANLN + NC inhibitor were 2.221 ± 0.188,0.769 ± 0.061,3.764 ± 0.371,2.265 ± 0.201,2.242 ± 0.018,and the difference among these groups was statistically significant (F =77.405,P <0.001),but the difference among psiCHECK-2ANLN group,psiCHECK-2-ANLN + NC group and psiCHECK-2-ANLN + NC inhibitor group was not statistically significant.However,luciferase activities of psiCHECK-2-ANLN + miR-217 group were significantly decreased when compared with other 3 groups,and luciferase activity of psiCHECK-2-ANLN +miR-217 inhibitor group were significantly increased when compared with other 4 groups (all P <0.001).Luciferase activities of groups transfected with psiCHECK-2-mutANLN was not significantly different (P =0.053).The expression of ANLN protein in PANC1 with psiCHECK-2-ANLN + miR-217 co-transfection was significantly down-regulated when compared with that with psiCHECK-2-ANLN transfection alone.Conclusions ANLN is one of the direct target genes of miR-217 in PANC1 cells.
9.Stable Surface Expression of a Gene for Helicobacter pylori Toxic Porin Protein with pBAD Expression System
PENG ZHIXIANG ; WEI XI ; LIN ZHENGMEI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(4):435-438
successive passages could express Hope protein, while only 1 from 5 E. coli colonies that contained lac operon-regulated plasmid encoding hopE gene could express HopE. Indi-rect immunofluorescence confirmed the expression of HopE on E. coli cell surface.
10.DNA vaccine encoding Streptococcus mutans surface protein protected gnotobiotic rats from caries.
Mingwen FAN ; Zhuan BIAN ; Zhixiang PENG ; Jihua GUO ; Rong JIA ; Zhi CHEN
Chinese Journal of Stomatology 2002;37(1):4-7
OBJECTIVEProtein of Streptococcus mutans is considered as one of the virulence factors due to its ability to mediate the initial attachment of Streptococcus mutans to tooth surface. In this study, an anticaries DNA vaccine pCIA-P was used to immunize rats. The expression of PAc in different tissues in vivo, specific immune response and protection effects against dental caries were observed.
METHODSPlasmid pCIA-P was injected into rats by two different routs: intramuscular injection (i.m.) and targeted salivary gland immunization (TSG). Immunohistochemistry technique was used to detect the expression of PAc. Gnotobiotic rats were vaccinated with pCIA-P by three different approaches: TSG, intramuscular injection and buccal mucosal injection (i.o.). The specific immune responses were evaluated by ELISA and their anticaries effects were evaluated by Keyes caries scores.
RESULTSPAc was expressed in the sarcoplasm and sarcolemma of muscle fibers and submandibular glands, especially strongly positive in duct regions. The levels of serum specific anti-PAc IgG and salivary specific anti-PAc IgA in TSG immunization and buccal mucosal immunization group were significantly higher than those of other groups. The Keyes caries scores of those two groups were significantly lower than those of other groups.
CONCLUSIONThe plasmid pCIA-P could provoke specific immune responses as a novel immunogen. Mucosal immunization with pCIA-P appears to be an effective genetic immunization method against dental caries.
Animals ; Antibodies, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Dental Caries ; prevention & control ; Germ-Free Life ; Immunization ; Male ; Membrane Glycoproteins ; Rats ; Rats, Wistar ; Streptococcal Vaccines ; immunology ; Streptococcus mutans ; immunology ; Vaccines, DNA ; immunology