Objective To establish a detection technique for H.pylori(HP) infection in Mongolian gerbils using nested PCR technique.Methods H.pylori was cultured in vitro and inoculated into Mongolian gerbils.At the 10th week after infection, the HP in the gastric juice of Mongolian gerbil was detected by conventional PCR assay and the gastric juice, gastric mucosa, duodenal contents and colon stool were examined by nested PCR.Rapid urease test and ELISA were used to analyze the accuracy of the nested PCR assay.All of the PCR products were verified by sequencing.Results The positive rate of gastric juice detected by conventional PCR was 30%, while the positive rates of gastric juice, gastric mucosa, duodenal contents and colon stool detected by nested PCR were 100%, 100%, 90%, and 10%, respectively.The positive detection rates of rapid urease test and serum ELISA were 100% and 0%, respectively.Comparing the results of different methods, both the positive rates of gastric juice and gastric mucosa detected by nested PCR and the detection rate of rapid urease test were 100%, but the results of conventional PCR detection of gastric juice, the nested PCR detection result of stool in colon and of serum ELISA assay were lower than other methods.Conclusions Due to its high accuracy and sensitivity, the nested PCR assay of gastric juice can be used for the long-time detection of H.pylori infection in Mongolian gerbils, especially useful in the experiments of prevention and treatment of H.pylori infection.

Objective To study the metabolic characteristics in type 2 diabetes mellitus with different BMI.Methods 233 patients with type 2 diabetes mellitus were divided into 2 groups. The first group was the BMI ＜25kg/m2 ,the second group with BMI ≥25kg/m2. The blood sugar、 blood-fat 、blood uric acid、type-B ultrasonic of liver were analyzed. Results The waistline、hip circumference、limosis c-peptide 、total cholesterol 、triglyceride and morbidity of fatty liver and hypertension of the second group were higher than the first group. Conclusion Over weigh patients of type 2 diabetes had IR、hyperlipemia、hypertension 、fatty liver. The rate and the severity of the artery diseases in type 2 diabetes mellitus increased wit the increase of the BMI. So weigh reduction could decrease the development of the artery diseases in type 2 diabetes.

Objective To study the effects of γ-IFN on the expression of HLA-G mRNA in primary cultured astrocytoma cells. Methods Different concentrations of γ-IFN were added to primary cultured cells, and HLA-G mRNA were detected by RT-PCR. Results After γ-IFN treatment, HLA-G mRNA can not be determined from HLA-G originally negative astrocytoma cells. The expression of HLA-G are up - regulated in all original HLA-G positive astrocytoma cells in a dose-dependent manner. Conclusion γ-IFN can increase the ex-pression of HLA-G gene in the primary cultured astrocytoma cells which HLA - G are originally positive.

Objective To investigate the infection and epidemiological characteristics of group A rotavirus (RV-A)and adenovirus in children with diarrhea in Guangzhou. Methods The colloidal gold technique was used to detect RV.A and adenovirus antigen in 2,171 stool samples from children with diarrhea in Guangzhou Women and Children′s Medical Center from January to December 2015,and the data were statistically analyzed. Results Among the 2,171 patients,the positive rate of RV-A infection was 17.96%and that of adenovirus infection 8.66%, and the co-infection rate of both virus was 3.45%. The positive rates between different genders were not significantly different(P > 0.05);the infectious time peak of RV-A was January(40.78%),followed by December(39.24%) and February(32.61%)and that of adenovirus infection was July(15.89%)and May(15.79%). The infectious peak of RV-A and adenovirus was December(7.29%),followed by January(7.01%). The peak age of infection ranged from 1y to 3y. Conclusion RV-A and adenovirus are the main pathogens of children diarrhea ,and the onset of virus infection has obvious seasonal change.

Objective To find out and clone the genes that can influence different willis circle in Meriones unguiculatus, the genomic library has been made.Methods 96 clean Meriones unguiculatus were dissected according to observe the differences of willis circle, and the mixed DNA was extracted from different blood vessels.The genomic library construction has been done by using pCC1FOS vector, following by CopyControl Fosmid Lib manual.The copy numbers, recombined segment size and recombination fraction of the library were measured.Results Genomic library of different Willis circle in Meriones unguiculatus was successfully made.The capacity of the library was 1700 copies.The segment size was 36kb, and the recombination fraction was 93%.Conclusion The library was the good beginning for the following steps which were gene cloning and genescreen.

BACKGROUND:Prolonged therapy with lamivudine has been associated with tyrosine-methionine-aspartate-aspartate mutation, which results in hepatitis B recurrence. Recently, antiviral agents, such as entecavir, have high efficacy and low resistance rate in hepatitis B-related liver disease. However, the researches on the effect of entecavir in preventing hepatitis B recurrence after liver transplantation are rare. OBJECTIVE:To investigate the effect of entecavir combined with low-dose hepatitis B immunoglobulin in preventing hepatitis B recurrence after liver transplantation. METHODS:The fol ow-up data of 253 patients who had liver transplantation for hepatitis B virus related liver disease were retrospectively analyzed. Al patients received nucleoside analogues therapy formal y before liver transplantation. The effects of entecavir+hepatitis B immunoglobulin and lamivudine+hepatitis B immunoglobulin were compared in al the patients and the patents with hepatitis B recurrence risk factors (positive preoperative HBeAg, DNA-positive hepatitis B virus, hepatoma and tyrosine-methionine-aspartate-aspartate mutation). RESULTS AND CONCLUSION:A total of 253 patients received hepatitis B virus-related liver transplantation, and 29 patients died. There were 202 patients in lamivudine group in which 26 patients were dead and 16 patients had hepatitis B virus recurrence, and the recurrence rate was 7.92%(16/202). However, entecavir group had 51 patients without hepatitis B virus recurrence in which three patients were dead. There were significant differences in the mortality rate and recurrence rate between two groups. Compared with the lamivudine+hepatitis B immunoglobulin, entecavir+hepatitis B immunoglobulin could effectively reduce the recurrence rate of the patients with hepatitis B virus-related risk factors. Hepatitis B immunoglobulin was terminated and nucleoside analogues were modulated when recurrence appeared. Al patients hepatitis B virus DNA were control ed less than 500 IU/mL and liver function returned to normal level. Log-rank test showed that there was no significant difference in the long-term survival rate after timely treatment of hepatitis B virus recurrence. With the prevention of nucleoside analogues combined with hepatitis B immunoglobulin therapy, timely treatment of hepatitis B recurrence has little influence on the prognosis. Entecavir combined with hepatitis B immunoglobulin can effectively prevent the hepatitis B recurrence. For the patients with hepatitis B virus-related risk factors, entecavir combined with hepatitis B immunoglobulin can better reduce the recurrence rate of hepatitis B than lamivudine+hepatitis B immunoglobulin after liver transplantation.

Objective:To investigate the antiviral efficacy of direct-acting antiviral agents (DAAs) in the treatment of liver transplantation (LT) recipients with hepatitis C virus (HCV) infection.Methods:Twenty-two HCV-infected LT recipients treated with DAAs at Fifth Medical Center of Chinese PLA General Hospital from December 2014 to June 2018 were retrospectively analyzed, Twenty cases of HCV RNA gene type 1b were treated with sofosbuvir (400 mg/d) + ledipasvir (90 mg/d) or sofosbuvir (400 mg/d) + daclatasvir (60 mg/d) for 12 weeks or 24 weeks; 2 cases of gene type 2a were treated with sofosbuvir (400 mg/d) for 12 weeks. The effect of antiviral treatment, adverse reactions during treatment, and laboratory indicators such as HCVRNA quantification, blood routine, liver and kidney function during treatment and follow-up were studied.Results:The LT recipients of HCV infection included 16 males and 6 females, with a median age of 61.5 (36-71) years old, and the median time of antiviral treatment was 48 (2-117) months after transplantation. Among the 22 patients, 16 received a 12-week course of treatment. Except for 2 patients who did not get HCVRNA negative conversion at 4-week, all achieved a negative HCV RNA at 4-week and the end of the treatment. Six LT recipients received a 24-week course of treatment (gene type 1b), and HCVRNA was negative at 4-week and the end of treatment. All patients achieved end of treatment virological response and a sustained virological response (SVR) rate of 100% at 12 weeks and 24 weeks after the end of treatment. The serum levels of alanine aminotransferase (ALT) and creatinine were 71.5 (30, 110) U/L and (89.4±25.7) mmol/L before treatment, respectively. ALT decreased to 22 (17.8, 28.5) U/L after 4 weeks of treatment, and serum creatinine decreased to (77.4±11.5) mmol/L at 24 weeks after the end of treatment. The differences before and after treatment were statistically significant (all P<0.05). No serious adverse events occurred during the treatment. Conclusions:DAAs have a definite antiviral effect in the treatment of LT recipients with HCV infection, and long-term SVR can be obtained.

ObjectiveTo demonstrate the efficacy and safety of Hangzhou tacrolimus capsule (Saishi Tac capsule,Hangzhou Zbongmei Huadong Pharmaceutical Co.Ltd,China) in Chinese liver transplant recipients.MethodsMulticenter,randomized open-labeled,prospective controlled clinical trial was performed in de novo Chinese liver transplant recipients.According to inclusive and exclusive criterion,83 liver recipients from 11transplant centers were enrolled.The recipients accepted Saishi Tac capsule,mycopheolate and steroid 48 h post-operation.The initial dose of Tac was 0.1-0.15 mg kg-1day-1and C0 was 8-12 ng/ml in the first 60 days,followed by 5-10 ng/ml until the terminal observation time poiut (12 weeks after transplantation).The efficacy and safety were estimated during the period.The primary efficacy endpoint of the study was the incidence of biopsy-confirmed acute rejection.Graft survival was the secondary endpoint.Safety was assessed by monitoring laboratory parameters and adverse events reported over the course of the study,such as infection,renal damage,hypertension,hyperlipema and diabetes mellitus and other adverse affairs.ResultsThe dose of Tac at 1st,2nd,4th and 8th week post-operation was (4.1±1.9),(4.5±2.1),(4.5±2.1),(4.4±1.8) and (4.1±2.1) mg,and correspondjng values to the C0 were (8.1±4.5),(8.9±4.5),(8.8±4.3),(8.8±4.1) and (8.0±2.8) ng/ml.During 12 weeks of follow-up,the incidence of biopsy-confirmed acute rejection was 4.8％ (4/83),and all of cases were reversed by implosive therapy.The survival rate of graft hver was 100％.The incidence of lung infection and diabetes mellitus was both 6.02％.ConclusionSaishi Tac capsule was safe and effective to Chinese liver transplant recipients.

Objective:To study the effect of Sanjie Xiaoliu Recipe on transplanted tumors in breast cancer mice through in vivo experiments, in order to explore the efficacy and mechanism of Sanjie Xiaoliu Recipe on breast cancer patients. Methods:45 BaL B/c female mice were selected to establish the transplanted tumor model of breast cancer. All the transplanted tumor models of breast cancer mice were randomly divided into five groups: the control group (intragastric administration of normal saline), the low, medium and high dose group of Sanjie Xiaoliu Recipe (intragastric administration of different doses of Sanjie Xiaoliu Decoction), and the paclitaxel group (intraperitoneal injection of paclitaxel). After 24 days of continuous administration, the diet and activities of mice were observed; the body weight, weight and volume changes of transplanted tumor mice were recorded, and the tumor inhibition rate was calculated. Western blot was used to detect the expression of epithelial mesenchymal transformation related proteins (E-cadherin, N-cadherin, Vimentin) in the transplanted tumor tissues of mice in each group.Results:(1) The food intake and activity status of mice treated with Sanjie Xiaoliu Recipe were less affected by the transplanted tumor of breast cancer. (2) The volume and weight of transplanted tumor in the treat groups were smaller than those in the control group (all P<0.01), and the volume of transplanted tumor in the middle dose group of Sanjie Xiaoliu Recipe was smaller than that in the low dose group ( P<0.05). The tumor inhibition rates among the treatment groups were: Sanjie Xiaoliu Recipe medium dose group 52.4%, paclitaxel group 40.3%, Sanjie Xiaoliu Recipe low dose group 39.5%, Sanjie Xiaoliu Recipe high dose group 34.1%. (3) The results of Western blot showed that the expression level of E-cadherin in the transplanted tumor tissue of the treat groups was higher than that in the control group, and the expression levels of N-cadherin and Vimentin were lower than that in the control group, with statistically significant difference (all P<0.05). Conclusions:The Sanjie Xiaoliu Recipe can improve the weakness and reduced consumption of breast cancer mice, which can inhibit the tumor mass growth in mice to a certain extent. Its mechanism may be that Sanjie Xiaoliu Recipe can inhibit the epithelial mesenchymal transformation of breast cancer and the invasion and metastasis of breast cancer.

ObjectiveTo investigate the risk factors for tumor recurrence and death after liver transplantation in patients with hepatocellular carcinoma (HCC) and their survival. MethodsThe patients with HCC who underwent liver transplantation in The Fifth Medical Center of Chinese PLA General Hospital from January 2005 to February 2019 were enrolled, and according to the presence or absence of HCC recurrence after liver transplantation, they were divided into recurrence group and non-recurrence group. The t-test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. Univariate and multivariate Cox proportional-hazards regression model analyses were used to determine the risk factors for HCC recurrence and death after liver transplantation. The Kaplan-Meier method was used for survival analysis, and the receiver operating characteristic (ROC) curve was used to investigate the predictive value of death-related risk factors after liver transplantation. ResultsA total of 391 HCC patients who underwent liver transplantation were enrolled, with a median follow-up time of 2 years, among whom 78(19.95%) experienced HCC recurrence. Preoperative alpha-fetoprotein (AFP) level＞200 ng/ml (recurrence: hazard ratio ［HR］=252, 95% confidence interval ［CI］: 1.58-4.03, P＜0.001; death: HR=2.99, 95%CI: 1.59-5.62, P＜0.001］, total tumor diameter (recurrence: HR=1.20, 95%CI: 1.12-1.28, P＜0.001; death: HR=1.10, 95%CI: 1.02-1.17, P=0.002), and vascular invasion (recurrence: HR=1.15, 95%CI: 1.04-1.26, P=0.016; death: HR=1.10, 95%CI: 1.03-1.18, P=0.004) were independent risk factors for tumor recurrence and death after liver transplantation. The 1-, 5-, and 10-year overall survival rates after liver transplantation were 94.8%, 84.2%, and 83.5%, respectively, and the 1-, 5-, and 10-year disease-free survival rates were 840%, 75.1%, and 75.1%, respectively. AFP, involvement of major blood vessels, body mass index, and total tumor diameter had a certain value in predicting the death of HCC patients with recurrence, with an area under the ROC curve of 0.789 (95% CI: 0.719-0858). ConclusionTumor biological features before transplantation are the key factors for tumor recurrence after transplantation.