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 investigate the association between hepatic fibrosis and anti-neutrophil cytoplasmic antibodies (ANCA)in the patients with schistosomiasis. Methods Totally 145 schistosomiasis patients confirmed by Jingzhou First Peo-ple's Hospital during the period of February 2014 to June 2016 were randomly selected as an experimental group,and 100 healthy people during the same period were randomly selected as a control group. The serum samples of the two groups were col-lected and the level of ANCA was detected by indirect immunofluorescence,and the levels of hyaluronic acid,laminin,procol-lagen type Ⅲ,and collagen type Ⅳ were detected by radioimmunoassay. The levels of the above indexes of the two groups were compared. Results The positive rate of ANCA was 28.96%(42/145)in the experimental group and 1.00%(1/100)in the healthy control group,and the difference between them was statistically significant(P < 0.05). The levels of hyaluronic acid, laminin,procollagen type Ⅲ and collagen type Ⅳ in the experimental group were significantly higher than those in the control group,and the differences were statistically significant(all P < 0.05),and the proportions of those with abnormal levels of hyal-uronic acid,laminin,procollagen type Ⅲ and collagen type IV in the experimental group were significantly higher than those in the control group,and the differences were statistically significant(all P < 0.05). In the experimental group,among the 42 pa-tients with ANCA positives,the proportions of those with abnormal levels of hyaluronic acid and laminin were significantly high-er than the proportions of those with ANCA negatives,and in the former,the levels of the four indicators of liver fibrosis were all higher than the normal values,and the average levels of hyaluronic acid and laminin in the patients with ANCA positives were significantly higher than those in the patients with ANCA negatives(all P < 0.05). Conclusion The patients with schistosomia-sis hepatic fibrosis could be positive for the autoimmune antibody ANCA,and ANCA may be associated with the autoimmune process of liver fibrosis.

Objective Explore a new treatment by orthogonal design of no empty cases :treatment of knee osteoarthritis by nerve locator ,for knee joint peripheral nerve branch implement selective radio frequency ablation .Methods According to patients′age ,body mass index ,gender ,organs ,in the course of the disease ,preoperative VAS score ,first divided into nine groups ,each group of four people ,then randomly assigned to four groups and each group 9 cases ,a total of 36 people .With nerve locator ,design of 4 factors and 3 levels ,no empty cases of orthogonal design ,every time 9 people by radio frequency ablation treatment of knee joint pe-ripheral nerve .Repeat 4 times to reduce the sampling error ,obtain the VAS difference between postoperative 1 week and preopera-tive .Results The best combination of treat knee joint osteoarthritis by Radio frequency ablation was A3B3C(1-3)D(1-3) .Selective radio frequency ablation knee peripheral nerve branch target rf temperature is 90 ℃ ,120 seconds ,lidocaine concentration(0 .5% to 2% ) and lidocaine measurement (0 .5 to 1 .5 mL) .Conclusion Using the orthogonal design of no empty cases ,for knee joint pe-ripheral nerve branch for selective radio frequency ablation by nerve locator .Treatment of knee joint osteoarthritis in A3B3C(1-3) D (1-3) combination effect exact ,minimally invasive ,safe ,accurate location ,the treatment effect is good ,low cost ,simple and high ef-ficiency .

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 .

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 observe the treatment effect of joint manipulation under brachial plexus anesthesia combined with comprehensive physical therapy in treatment of periarthritis of shoulder. Mehods From 2010.3 to 2010.9, 60 patients with periarthritis of shoulder in the Department of Pain, The First People's Hospital of Jingzhou,were divided into 3 groups:group A,B and C.Twenty patients in Group A were given Joint manipulation under brachial plexus anesthesia for treatment of the periarthritis of shoulder. Twenty patients in Group B were given comprehensive physical therapy fro treatment of periarthritis of shoulder. Twenty patients in Group C were given Joint manipulation under brachial plexus anesthesia combined with comprehensive physical therapy for treatment of periarthritis of shoulder.Results All 60 patients were followed up for 1 months. At the 1st month after surgery, vas score was evaluated for patients. Postoperative and preoperative vas score difference was compared between 3 groups by t test. There was statistically significant difference in the postoperative and preoperative vas score between Group C and Group A,and between Group B and Group A (P<0.05) . Conclusion The treatment effect of Joint manipulation under brachial plexus anesthesia combined with physical therapy in treatment of periarthritis of shoulder is good, but has no obvious difference compared with comprehensive physical therapy.

Objective To observe the treatment effect of radio frequency target combined with collagenase in treatment of cervical intervertebral disc herniation. Methods From March 2011 to September 2011, 40 patients with cervical intervertebral disc herniation were divided into 2 groups: Group A and Group B. Patients in Group A were given radio frequency target radiofrequency for treatment of the cervical intervertebral disc herniation and patients in Group B of 20 were given radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation. Results 40 patients were followed up for 6 months. At the 1st week and 6 months after surgery, the VAS score difference between before and after surgery in patients were compared between two groups by t test. There was no statistically significant difference at the 1st week after surgery between two groups (P>0.05), while 6 months after surgery, there was statistically significant difference between two groups (P<0.05) .Conclusion Radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation can alleviate the pain symptoms of patients,and the short-term and long-term curative effect is very good. The long-term curative effect has statistically significant difference with pure target radiofrequency ablation treatment. Radio frequency target combined with collagenase for treatment of cervical intervertebral disc herniation has good safety, few side effects, low risks and good short-term and long-term curative effect, so it deserves to be generalized.