1.Expression and clinical meaning of nm23-H1 gene in gastric carcinoma tissue
Kai LU ; Zhuangkai YAO ; Yanwen LIU ; Cheng ZHAO ; Yongping LIU
Journal of Endocrine Surgery 2009;3(2):88-90
Objective To study the relationship of expression of metastasis suppressor gene nin23-H1 protein in gastric carcinoma tissue and tumor infiltration and metastasis.Methods The expression of metastasis suppressor gene nm23-H1 protein in 50 gastric carcinoma tissues was detected by SP immunohistocbemistry tech-nology.Results Correlations were presented among expression of nm23-H1 protein, infiltrate grade of gastric carcinoma, lymph node metastasis and TNM stage.Conclusions Expression of nm23-H1 may suppress infiltra-tion and metastasis of gastric carcinoma, Low expression of nm23-H1 protein may be of great reference value in judgement of tumor progression, metastasis and reeidivation.
2.Expression and clinical significance of nm23-H1,c-Met and survivin protein in gastric carcinoma tissue
Kai LU ; Zhuangkai YAO ; Yanwen LIU ; Cheng ZHAO ; Yongping LIU
Journal of Endocrine Surgery 2009;3(4):226-229
Objective To Study the expression and clinical meaning of nm23-H1,c-Met and survivin protein in gastric carcinoma tissue.Methods Expression of nm23-H1,c-Met and survivin protein in 50 paraffin block samples of gastric carcinoma were detected by S-P immunohistochemistry technology.The relationship of expression to tumor differentiation degree,infiltration depth,lymph node metastasis,TNM stage,tumor metastasis and reeidivation were analyzed.Results The positive expression rates of nm23-H1,c-Met and survivin protein were 46%,72% and 80%,low nm23-H1 protein expression and high c-Met protein expression correlated tightly with TNM stage and lymph node metastasis of gastric carcinoma,high survivin protein expression correlated tightly with differentiation degree,TNM stage and lymph node metastasis of gastric carcinoma,there was a inverse correlation between nm23-H1 protein expression and survivin,but a direct correlation between c-Met and survivin.Conclusions Low nm23-H1 protein expression and high c-Met and survivin expression has important guidance significance of making a early diagnosis and judging prognosis.Selective gene therapy guided by these may be helpful.
3.Effect of hydrogen sulfide postconditioning on systolic function of left ventricle during myocardial ischemia-reperfusion in rats
Chi CHENG ; Tian YU ; Xingkui LIU ; Shengli DENG ; Gang YAO
Chinese Journal of Anesthesiology 2010;30(10):1175-1178
Objective To investigate the effect of hydrogen sulfide postconditioning on the systolic function of left ventricle during myocardial ischemia-reperfusion (IR) in rats. Methods Part Ⅰ Adult male SD rats weighing 200-250 g were anesthetized with pentobarbital 40 mg/kg and heparin 250 U/kg. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95% O2-5% CO2 at 37 ℃. Forty isolated rat hearts were randomly divided into 5 groups ( n = 8 each) after 20 min of equilibration: control group (group C); IR group; sodium hydrosulfide 1,10, 100 μmol/L postconditioning group (group SP1, SP10, SP100 ).In group Cthe hearts were perfused continuously for another 100 min. In group IR, the hearts were reperfused for 60 min after 40 min ischemia induced by 10 ml/kg ST. Thomas solution. In group SP1 , SP10 and SP100 the hearts were perfused with K-H solution containing sodium hydrosulfide 1, 10, 100 μmol/L for 2 min before reperfusion.LVDP and ± dp/dtmax were recorded at the end of equilibration and reperfusion. Part Ⅱ Cardiomyocytes were isolated from the male SD rats (weighing 200-250 g) and then cultured in CO2 incubator for 4 h. Sixty-four dishes of cultured myocytes were randomly divided into 4 groups( n = 16 each): control group (group C), hypoxia/reoxygenation group (group HR), hydrogen sulfide postconditioning group (group SP) and hypoxia postconditioning group (group HP). Group C were cultured continuously for 2 h. Group HR, SP and HP were exposed to 1 h hypoxia (95%N2-5%CO2 ) followed by 1 h reoxygenation. In group SP 10 μmol/L sodium hydrosulfide was added and the myocytes were then incubated for 2 min before reoxygenation. In group HP the cultured myocytes were expased to 3 min reoxygenation followed by 3 min hypoxia for 3 times before the 1 h reoxygenation. Mitochondrial membrane potential and F-actin expression were determined. Results Part Ⅰ Compared with group C, LVDP and ± dp/dtmax were significantly decreased at the end of reperfusion in group IR (P < 0.05), while no significant difference was found in group SP1 , SP10 and SP100(P >0.05). Compared with group IR, LVDP and ± dp/dtmax were significantly increased in group SP ( P < 0.05). There was no significant difference in LVDP and ± dp/dtmax among group SP1, SP10 and SP100(P >0.05). Part H Compared with group C, the mitochondrial membrane potential was significantly decreased in group HR and HP, and the expression of F-actin was significantly up-regulated in group HR, SP and HP ( P < 0.05). Compared with group HR, the mitochondrial membrane potential was significantly increased and the expression of F-actin up-regulated in group SP and HP ( P < 0.05 ). There were no significant difference in the mitochondrial membrane potential and expression of F-actin between group SP and HP ( P >0.05).Conclusion Hydrogen sulfide postconditioning can improve left ventricular systolic function during IR in rats by stabilizing mitochondrial membrane potential and promoting aggregation of F-actin.
5.Th cell differentiation in chronic stage of viral myocarditis in mice and interference of Qingxin-II Recipe
Zhiqing CHENG ; Hongfei LIU ; Juan ZHANG ; Hangping YAO
Journal of Integrative Medicine 2007;5(3):318-21
OBJECTIVE: To investigate part mechanisms of Th cell differentiation, and to observe the interference effect of Qingxin-II Recipe in the chronic stage of viral myocarditis (VMC), so as to provide some experimental evidences for illuminating the pathogenesis of VMC and treatment mechanisms of Qingxin-II Recipe. METHODS: According to 20%-40% death rate of experiment in advance, 100 BALB/c male mice (4 weeks old and weighing 12-15 g) were used. Twenty mice were randomly assigned to normal control group, and the other 80 mice were intraperitoneally injected with 0.1 ml normal saline containing coxsackie virus B3 at the 1st, 4th and 28th day (the virus densities were 1:2000, 1:1000 and 1:500 respectively) to induce chronic VMC. At the 42nd day, the surviving mice were randomly divided into untreated group and treatment group, with 20 mice in each group. Mice in the treatment group were orally administered with 0.2 ml Qingxin-II Recipe every day, while mice in the normal control group and the untreated group were administered with 0.2 ml normal saline. All the mice were sacrificed after 45 days, and the sera, heart and spleen cells were collected. Then the myocardial pathological changes were observed by using a light microscope, and the levels of IL-4, IL-10 and IFN-gamma in serum were detected by enzyme linked immunosorbent assay (ELISA). And the Th cell differentiation was observed by flow cytometry. RESULTS: No obvious myocardial pathological changes were observed in mice of the normal control group. Myocardial pathological changes in the treatment group were slighter than those in the untreated group. The difference of serum IL-10 level between the normal control group and the untreated group showed no significance (P>0.05), and the levels of IFN-gamma and IL-4 of the untreated group were higher than those of the normal control group (P<0.05 or P<0.01). There was no statistical difference in IL-10 level between the treatment group and the untreated group (P>0.05), while the serum levels of IL-4 and IFN-gamma of the treatment group were lower than those of the untreated group (P<0.05 or P<0.01). There was no significant difference of the Th1 cell responder between the treatment group and the untreated group (P>0.05), while the Th2 cell responder was inhibited significantly in the treatment group (P<0.05). CONCLUSION: Qingxin-II Recipe can restore the balance of Thl and Th2 cells through inhibiting the reaction of Th2.
6.Investigation of Human Papillomavirus-16 Infections and Its Mixed Infection in Yunnan Region
Zheng LI ; Si CHENG ; Lei SHI ; Yufeng YAO ; Feng LIU
Journal of Kunming Medical University 2014;(1):92-94
Objective To investigate the distribution of Human Papillomavirus 16 (HPV16) infection and its mixed infection with other HPV subtypes in the Yunnan region. Methods 16 166 cases of women were tested using flow fluorescence Luminex technology. Results (1) HPV16 infection rate and mixed infection rate was 2.2%and 28.0%, respectively; (2) The most common type of HPV16 mixed infection was HPV52, followed by HPV33. The two kinds of mixed infection accounted for 39.8% of the total infection rate; (3) There was a significant difference between each age group of HPV16 mixed infection (Chi=26.39, <0.01) . Conclusion The HPV16 infection was mainly HPV infection in Yunnan region. HPV16 mixed infection merged mainly with HPV52 and HPV33. HPV16 mixed infection was associated with age.
7.Construction of stable Cdc25C knockout HeLa cell strains using CRISPR/Cas9 gene-editing system
Yebao YAO ; Guangfei WANG ; Qingcai DONG ; Cheng CAO ; Xuan LIU
Military Medical Sciences 2017;41(5):359-362
Objective To knockout the cell division cycle 25 homolog C(Cdc25C) gene in HeLa human cervical cancer cells and to construct HeLa Cdc25C gene knockout stable strains using clustered regularly interspaced short palindromic repeats(CRISPR)/Cas9 gene-editing system.Methods The sequence of the small guide RNA(sgRNA),which could specifically recognize the first exon of Cdc25C,was designed according to the target-designing rules of CRISPR/Cas9 for construction of eukaryotic recombinant expressional plasmids.After sequencing,the plasmid was transfected into HeLa cells.The stable Cdc25C-knocking out strains were screened through the stress of puromycin,and the knockout effect was detected by Western blotting.The cell cycle was analyzed by flow cytometry.Results The stable Cdc25C-knocking out strains were obtained.Moreover,the gene′s knockout obviously delayed the progression of G2/M phase.Conclusion The HeLa Cdc25C gene knockout stable strain is successfully built using CRISPR/Cas9 system,facilitating studies on the function of Cdc25C and the mechanism of carcinogenesis.
8.Preparation and characterization of composite delivery system of paclitaxel-loaded temperature sensitive liposome and siRNA-loaded gold nanostar
Ziting CHENG ; Ke YAO ; Wanwan LIU ; Bohui XU ; Hongyan ZHU
Journal of China Pharmaceutical University 2017;48(4):445-452
We prepared gold nanostar (GNS) through seed growth method firstly,then formation of COX-2 siRNA(siCOX-2) and GNS composite modified with polyethylene glyco (PEG),2-amino-2-deoxy-D-glucos (DG) and 9-D-arginin (9R) was prepared.Mterwords,paclitaxel temperature sensitive liposome (PTX-TSL) was prepared by film dispersion method.Finally,siCOX-2 delivery systerm (PTX-TSL-(siCOX-2(9R/DG-GNS)))was obtained by hydrosulfuryl ligand reaction between siCOX-2 (9R/DG-GNS) and PTX-TSL The successful build of siCOX-2 (9R/DG-GNS) was vetified by nuclear magnetic resonance (NMR),sodium dodecyl sulfate polyacryl amide gel electrophoresis (SDS-PAGE),and ultraviolet spectrophotometry and agarose gel electrophoresis method.Particle size of PTX-TSL-(siCOX-2(9R/DG-GNS)) was (292 ± 14) nm and Zeta potential was about -(2.59 ± 0.12) mV,which were measured by Zetasizer Nano ZS90.The morphology of PTX-TSL-(siCOX-2 (9R/DG-GNS)) measured by transmissionelectronmicroscopy showed homogeneous star structure with phospholipid bilayer on the surface,and it showed good thermal conversion efficiency under radiation of 808 nm laser.Differential scanning calorimetry test showed that PTX-TSL phase transition temperature is about 42.6 ℃.The drug loading content(using dialysis method) and encapsulation efficiency of PTX-TSL were about 7.5% and 95.4%,at the same time,the release process experiment of PTX-TSL showed that it had a good temperature sensitive release performance.It is hopeful that this siCOX-2 system can be used for reducing drug resistance of PTX and improving the treatment effect of PTX through the synergistic antitumor drug resistance effect of siCOX-2.
9.Safety and effectiveness of ozone versus hyaluronic acid for treating knee osteoarthritis:a Meta analysis
Bingyan HU ; Jinwei AI ; Chufan LIU ; Qiong CHENG ; Zhongjun YAO
Chongqing Medicine 2017;46(9):1240-1245,1248
Objective To compare the safety and effectiveness between ozone (O3) and hyaluronic acid (HA) in treating knee osteoarthritis(KOA) by using the meta analysis method.Methods The relevant randomized controlled trials(RCTs) in PubMed,Cochrane Library (issue 1,2016),Embase,CNKI,CBM,VIP,and Wan-Fang databases were retrieved from their establishment to January 23,2016.Two reviewers independently screened the literatures,extracted the data and evaluated the quality of the included RCTs.The results were performed the statistical analysis by using the RevMan5.3 and Stata13.0 software.Results Twenty RCTs involving 2 136 KOA patients were included.Compared with the HA treatment of KOA,the O3 treatment had higher treatment effective rate[odds ratio(OR) =2.78,P<0.01],and better pain relief effect[at 1,3,6 month after treatment:mean difference(MD) =-0.25,-0.71,-1.70,P<0.01].There were no statistically significant differences in complications between the two treatment methods[OR=0.84,P=0.56].Conclusion Current evidences indicate that the short-term therapeutic effect of O3 for KOA is superior to HA,and the safety is similar.
10.Location and expression of vitamin D receptor in the duodenum of C57BL/6 mice at different developmental stages
Jia CHENG ; Shaowei LIU ; Wanying CAO ; Yiqi YANG ; Wang YAO
Acta Laboratorium Animalis Scientia Sinica 2017;25(3):320-324
Objective To investigate the relationship of vitamin D with the intestinal development and study the expression of vitamin D receptor (VDR) in the duodenum of C57BL/6 mice at different developmental stages.Methods Quantitative PCR (qPCR),histology using H&E staining and immunofluorescence staining,and Western blotting (WB) were performed to elucidate the expression of VDR in mice intestine at different growth and developmental stages.Results The peak of VDR mRNA expression reached on 21 d.The pathological result showed that VDR mainly distributed in the cytoplasm of epithelial cells and smooth muscle cells in the mouse duodenum.WB result indicated that there was no nuclear translocation of VDR protein in the mouse duodenum.Conclusions This study demonstrates the regularity of expression of VDR in the mouse duodenum during its development,and contributes to understanding the function of VDR in the intestines.