Objective:To explore the effect of circ_0101145 on immune escape of gastric cancer(GC)cells through miR-548c-3p/programmed death-ligand 1(PD-L1)axis.Methods:RT-qPCR and Western blot were used to detect circ_0101145,miR-548c-3p,PD-L1 mRNA and protein levels in GC tissue and cell lines,the relationship between circ_0101145 level and clinical patho-logical parameters of patients was analyzed;dual Luciferase experiment was used to verify the regulatory relationship of circ_0101145 on miR-548c-3p,miR-548c-3p on PD-L1;liposome transfection method was used to structure BGC-823 transfected cell line,divided into circ-NC group,circ_0101145 group,si-circ-NC group,si-circ_0101145 group,circ_0101145+NC group,circ_0101145+miR-548c-3p group,miR-548c-3p+NC group,miR-548c-3p+PD-L1 group.CD8+T cells were separated from human peripheral blood and cocultivate with BGC-823 cell;flow cytometry was used to detect CD8+T cells apoptosis rate;ELISA was used to detect TNF-α,IFN-γ levels in coculture medium;CCK-8,EdU staining,Transwell assay were used to detect cell proliferation,invasion ability.Nude mice was subcutaneous inject with BGC-823 cell suspension of knockdown circ_0101145,after 30 days,growth of transplanted tumors were measured.Results:Compared with adjacent cancer tissues or normal cells,miR-548c-3p level in GC tissues and cell lines were de-creased,while circ_0101145,PD-L1 mRNA and protein levels were increased,and the higher circ_0101145 level,the higher TNM stage,the lower degree of tumor differentiation,the easier microvascular infiltration,and the easier lymph node metastasis(P<0.05);circ_0101145 targeted miR-548c-3p,miR-548c-3p targeted PD-L1.In the cocultivation system,overexpression of circ_0101145 could increase CD8+T cells apoptosis rate,and TNF-α,IFN-γ levels in coculture supernatant were reduced,the A value after 1 day,2 days and 3 days,EdU positive rate and the number of invasive cell were increased(P<0.05).Knock down of circ_0101145 could de-crease the apoptosis rate of CD8+T cells,and TNF-α,IFN-γ levels in coculture supernatant were increased;the A value after 1 day,2 days and 3 days,EdU positive rate and the number of invasive cell were decreased(P<0.05).Overexpression of miR-548c-3p could partially reverse the effect of overexpression of circ_0101145 on the above indicators(P<0.05).Overexpression of PD-L1 could partially reverse the effect of overexpression of miR-548c-3p on the above indicators(P<0.05).Knock down of circ_0101145 could inhibit the growth of transplanted tumors in mice(P<0.05).Conclusion:circ_0101145 promotes immune escape of GC cells through miR-548c-3p/PD-1 axis.

Objective:To explore the effect of circ_0101145 on immune escape of gastric cancer(GC)cells through miR-548c-3p/programmed death-ligand 1(PD-L1)axis.Methods:RT-qPCR and Western blot were used to detect circ_0101145,miR-548c-3p,PD-L1 mRNA and protein levels in GC tissue and cell lines,the relationship between circ_0101145 level and clinical patho-logical parameters of patients was analyzed;dual Luciferase experiment was used to verify the regulatory relationship of circ_0101145 on miR-548c-3p,miR-548c-3p on PD-L1;liposome transfection method was used to structure BGC-823 transfected cell line,divided into circ-NC group,circ_0101145 group,si-circ-NC group,si-circ_0101145 group,circ_0101145+NC group,circ_0101145+miR-548c-3p group,miR-548c-3p+NC group,miR-548c-3p+PD-L1 group.CD8+T cells were separated from human peripheral blood and cocultivate with BGC-823 cell;flow cytometry was used to detect CD8+T cells apoptosis rate;ELISA was used to detect TNF-α,IFN-γ levels in coculture medium;CCK-8,EdU staining,Transwell assay were used to detect cell proliferation,invasion ability.Nude mice was subcutaneous inject with BGC-823 cell suspension of knockdown circ_0101145,after 30 days,growth of transplanted tumors were measured.Results:Compared with adjacent cancer tissues or normal cells,miR-548c-3p level in GC tissues and cell lines were de-creased,while circ_0101145,PD-L1 mRNA and protein levels were increased,and the higher circ_0101145 level,the higher TNM stage,the lower degree of tumor differentiation,the easier microvascular infiltration,and the easier lymph node metastasis(P<0.05);circ_0101145 targeted miR-548c-3p,miR-548c-3p targeted PD-L1.In the cocultivation system,overexpression of circ_0101145 could increase CD8+T cells apoptosis rate,and TNF-α,IFN-γ levels in coculture supernatant were reduced,the A value after 1 day,2 days and 3 days,EdU positive rate and the number of invasive cell were increased(P<0.05).Knock down of circ_0101145 could de-crease the apoptosis rate of CD8+T cells,and TNF-α,IFN-γ levels in coculture supernatant were increased;the A value after 1 day,2 days and 3 days,EdU positive rate and the number of invasive cell were decreased(P<0.05).Overexpression of miR-548c-3p could partially reverse the effect of overexpression of circ_0101145 on the above indicators(P<0.05).Overexpression of PD-L1 could partially reverse the effect of overexpression of miR-548c-3p on the above indicators(P<0.05).Knock down of circ_0101145 could inhibit the growth of transplanted tumors in mice(P<0.05).Conclusion:circ_0101145 promotes immune escape of GC cells through miR-548c-3p/PD-1 axis.

Objective:To explore the effect of long noncoding RNA(lncRNA)THAP7-AS1 on the glycolysis of gastric cancer(GC)cells by regulating methyltransferase-like 3(METTL3)mediated N6-methyladenosine(m6A)modification. Methods:Gene Expression Profiling Interactive Analysis(GEPIA)database was used to analyze the expression levels of THAP7-AS1 and METTL3 in GC tissues and their relationship with the overall survival of GC patients.Real-time fluorescence quantitative PCR and Western blotting were used to analyze the expression of THAP7-AS1,METTL3 mRNA,glucose transporter 1(GLUT1)mRNA,and METTL3 protein in GC tissues and paracancerous tissues samples collected from 80 GC patients in Department of Oncology,The Third Affiliated Hospital of Zunyi Medical University(the First People's Hospital of Zunyi),and the relationship between THAP7-AS1 levels and the clinicopathological characteristics of GC patients was analyzed.Real-time fluorescence quantitative PCR and Western blotting were used to verify the expression of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA,and METTL3 protein in GES-1,BGC-823 and SGC-7901 cells.Lentiviral infection was used to knock-down THAP7-AS1 or overexpress METTL3 BGC-823 and SGC-7901 cells,and real-time fluorescence quantitative PCR and Western blotting were used to examine effect of different treatment on the expression of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA,and METTL3 protein;colorimetry assay was used to detect the m6A modification level in the total RNA;methylated RNA immunoprecipitation(MeRIP)-quantitative PCR(qPCR)was used to detect the GLUT1 m6A modification level;glycolysis stress test kits were used to detect the extracellular acidification rate(ECAR),glucose uptake and lactate production of treated GC cells;Western blotting was used to examine the expression levels of METTL3,GLUT1,M2 type pyruvate kinase(PKM2)and lactic dehydrogenase(LDHA)proteins in treated GC cells;EdU staining,wound healing assay and Transwell invasion assay were used to evaluate the proliferation,migration and invasion of treated GC cells.Finally,a mouse model of subcutaneously transplanted GC tumor was established using nude mice,and the effect of knocking-down THAP7-AS1 was assessed by measuring the tumor volume and weight,as well as the expression levels of METTL3 and GLUT1 proteins in transplanted GC tumor tissues. Results:Analysis of the GEPIA database showed that the expression levels of THAP7-AS1 and METTL3 was higher in GC tissue than those in normal gastric tissues,and the expression levels of THAP7-AS1 and METTL3 are negatively correlated with overall survival of GC patients(P＜0.05).Compared with the paracancerous tissues(or normal gastric epithelial cells),the expression levels of THAP7-AS1,METTL3 mRNA,GLUT1 mRNA and METTL3 protein was significantly increased in GC tissues(or GC cells),and the higher the expression of THAP7-AS1,the higher the TNM stage,the lower the degree of tumor differentiation,and the easier the occurrence of microvascular infiltration and lymph node metastasis(P＜0.05).Knocking-down of THAP7-AS1 down-regulated the expression levels of METTL3 mRNA,GLUT1 mRNA and METTL3 protein,the m6A modification levels in total RNA and GLUT1,the ECAR levels,the glucose uptake,the lactate production,EdU positive rate,scratch healing rate,the number of invaded cells,and the expression levels of glycolysis-related proteins(METTL3,GLUT1,PKM2 and LDHA)in GC cells(P＜0.05).Overexpression of METTL3 could partially reverse these effects of THAP7-AS1 knock-down(P＜0.05).In vivo experiments showed that THAP7-AS1 knock-down can obviously inhibit the growth of transplanted GC tumors(P＜0.05). Conclusion:lncRNA THAP7-AS1 can promote the glycolysis which further promotes the proliferation,migration and invasion of GC cells by regulating METTL3 mediated m6A modification.

Objective To evaluate the role of emergency and staged hepatectomy in peritoneal metastasis associated with ruptured hemorrhage of resectable hepatocellular carcinoma (HCC) patients,and investigate the impact of surgery timing-selecting on peritoneal metastasis of postoperative HCC patients.Methods A retrospective analysis was conduct on the pooled data from 38 HCC patients with spontaneously ruptured hemorrhage treated in our hospital from August 2011 to January 2016.These patients were divided into emergency group who underwent hepatectomy within 24 hours at admission,and staged group who underwent the procedure one week after admission.Perioperative events,overall survival (OS) and disease-freesurvival (DFS) rates,incidence of recurrent and metastatic disease were compared between the two groups.Results The perioperative blood loss and transfusion were much more in emergency group than staged group (both P ＜ 0.05).Nevertheless,the incidence of postoperative mortality was not significantly different (6.0％ vs 0％,P ＞ 0.05).The median survival was 22.5 months in emergency group versus 14.2 months in staged group.The 6-month,1-year,3-year OS rates in emergency group were 88.2％,82.4％ and 30.3％ respectively,and 6-month,1-year,3-year DFS rates were 81.3％,54.7％ and 27.3％.The 1-year OS and 6-month DFS rates were higher than those of staged group (both P ＜ 0.05).The incidence of peritoneal metastasis in staged group was higher than that in emergency group,but it was not significantly different (38.1％ vs 29.4％,P ＞ 0.05).Univariate and multivariate analysis indicated that tumor diameter ≥ 10 cm and AFP ＞ 10 000 μg/L were the risk factors for peritoneal metastasis after hepatectomy for HCC patients with spontaneously ruptured hemorrhage.Conclusions Emergency hepatectomy would warrant a better short-term prognosis compared with staged hepatectomy for the HCC patients with spontaneously ruptured hemorrhage.Staged hepatectomy would not raise the possibility of postoperative peritoneal metastasis.The predictors of tumor diameter ≥ 10 cm and AFP ＞ 10 000 μg/L were risk factors for peritoneal metastasis after hepatectomy for spontaneously ruptured HCC patients.

Objective To explore a variety of levels of serum marker test applications in the diagnosis of fatty liver .Methods Data were randomly selected from April 2013 to April 2014 for treatment of patients with fatty liver hospital 45 cases ,set the study group ,choose the same period in healthy volunteers to undergo a medical examination in our hospital 45 cases ,it was set to control group ,two groups of subjects were taking a variety of levels of serum markers tested .Comparison and analysis of two groups of subjects to detect a variety of levels of serum markers and positive case detection rate .Results The study group subjects ALT , AST ,TG ,TC index the average level of detection was higher than the control group ,statistically significant differences (P<0 .01);study group subjects ALT ,AST ,TG ,TC index the positive rates were 77 .78% ,93 .33% ,55 .56% ,46 .67% more than 8 .89% in the control group ,4 .44% ,15 .56% ,11 .11% higher ,statistically significant differences (P<0 .05);United biochemical indicator de‐tection of biochemical indicators of detection rate of fatty liver was obviously higher than that of single detection rate ,the difference was statistically significant (P<0 .05) .Conclusion Multiple levels of serum markers of fatty liver diagnostic test in higher detec‐tion rate .

Objective To explore the value of comprehensive biochemical and serological test in Salmonella clinical test .Methods A total of 95 cases of suspected salmonella infection with enteric fever were selected as subjects in this study from April 2013 to A‐pril 2014 ,and 48 cases randomly divided in research group ,47 cases in control group .Regular inspection was conducted in the con‐trol group ,comprehensive biochemical and serological test were conducted in the control group .Compared the results in the two groups .Results The salmonella detection rate in the research group was 93 .75% ,which was significant higher than 68 .09% in the control group (P< 0 .05) .Conclusion Combined detection of comprehensive biochemical and serology test could detect Salmonella effectively ,it is worthy of application and popularization in clinic .

Salmonella Typhi capsular polysaccharide vaccines were encapsulated in the Micro-particles made from polyethylene glycol-poly-DL-lactide (PELA). BALB/c mouse were divided into three groups with 20 mice in each. Mouse were immunized respectively with controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines and Salmonella Typhi capsular polysaccharide vaccines by oral and subcutaneous administration. The mice blood and salvia were collected at the 2nd, 4th and 8th weeks respectively for the titrating of IgG and sIgA antibodies by RIA. At the 8th week, live typhoid bacteria were injected into the immunized mice for the calculation of the rate of immunization protection. The IgG titers of the controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines group were higher than those of the other groups(P < 0.05). The IgA titers of the low groups of controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines (oral and subcutaneous) were higher than those of the group of Salmonella Typhi capsular polysaccharide vaccines (P < 0.05). The immunization protection rates of the three groups were 40%, 100% and 60% respectively. The controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines possess the advantages of releasing slowly in vivo and persisting long time immunogenicity.
Administration, Oral ; Animals ; Delayed-Action Preparations ; Female ; Immunoglobulin A, Secretory ; analysis ; Immunoglobulin G ; blood ; Injections, Subcutaneous ; Mice ; Mice, Inbred BALB C ; Microspheres ; Polysaccharides, Bacterial ; administration & dosage ; immunology ; Typhoid-Paratyphoid Vaccines ; administration & dosage ; immunology ; Vaccination