Objective To detect the integration of hepatitis B virus (HBV) DNA in HBVrelated human hepatocellular carcinomas (HCC). Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction (PCR) with specially designed U-base primers. According to the known genes and human Alu repeat sequences (Alu repeat) , primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies.Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information (NCBI) basic local alignment search tool (BLAST) and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn't show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

ObjectiveTocomparethedifferencesoftwostocksofguineapigs,thealbinoguineapigsandpigment guinea pigs , in establishing dyslipidemic model , to evaluate their lipid-lowering action , and to compare their properties for development of hyperlipidemia .Methods Two stocks of the 5-week-old guinea pigs were randomly divided into two groups, normal group (NC) and model group (Model).For the NC group, 12 guinea pigs were fed with normal chew .For the model group , after fed with high-fat diet for four weeks , 24 male guinea pigs were randomly grouped and treated with vehicle (VC group) and pitavastatin (Pit group) calcium, respectively, by gavage as well as received high-fat diet.Before and after modeling and pitavastatin treatment , blood samples were collected and subjected to analysis of plasma TC , TG, HDL-C and LDL-C, respectively .Results In the normal group , the blood lipid levels of albino guinea pigs were more stable than that of the pigmented pigs with the increase of age .After fed with high-fat diet , the plasma lipid levels of TC , TG and LDL-C were significantly increased in the two strains of guinea pigs , while HDL-C showed a decrease to varying degrees .Interestingly , the lipid level in the albino guinea pigs was significantly higher than that of pigment guinea pigs . And also, after drug administration for four weeks , pitavastatin treatment significantly decreased the elevated lipid level of TC, TG and LDL-C in the albino guinea pigs compared with that in the pigment guinea pigs .Conclusions The albino guinea pigs and pigment guinea pigs demonstrate certain differences in establishing dyslipidemic model and evaluating lipid -lowering pharmacodynamics .However , compared with the pigment guinea pigs , the albino guinea pigs have obvious superiority because of easy establishment of hyperlipidemia model and are more sensitive to lipid -lowering drugs .

Objective To study the methylation status of secreted frizzled-related protein (SFRP) 1 and SFRP2 genes in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) and the relationship between the methylation status of the two genes and the development of HCC.Methods Using methylation-specific polymerase chain reaction (MSP) to detect methylation status of SFRP1 and SFRP2 genes of 45 specimens of HCC tissue and adjacent non-tumorous liver tissue from HCC patients during operations,and 6 normal liver tissues from patients with cholecystolithiasis or hepatic hemangiomas. The data were analyzed by chi-square test and Fisher exact test. Results SFRP1 gene methylation was detected in 28 HCC tissues and 16 adjacent non-tumorous liver tissues,accounted for 62.2% and 35.6%,respectively;and SFRP2 gene methylation was detected in 23 HCC tissues and 13 adjacent non-tumorous liver tissues,accounted for 51.1% and 28.9%,respectively;while no methylation was detected in 6 samples of normal liver tissues. There was no significant difference between the methylation of SFRP1 and SFRP2 genes in HCC tissues and gender,age,HBV serum markers,types of adjacent non-tumorous liver tissues,metastasis and pathological stage (P＞0.05).The abnormal methylation status between SFRP1 and SFRP2 genes was linear correlated in HCC tissues (r=0.381,P=0.01).Conclusion Hypermethylation of SFRP1 and SFRP2 genes frequently occurs in HBV-related HCC,which may be an important molecular biomarker for prediction of hepatocarcinogenesis in the future.

Objective To analyze the correlation between covalently closed circular DNA (cccDNA) in the peripheral blood mononuclear cells (PBMC) of hepatitis B virus (HBV)-infected patients and serum HBV DNA,hepatitis B surface antigen (HBsAg),hepatitis B e antigen (HBeAg) and liver histology of hepatitis B patients,and to explore the clinical significance of HBV cccDNA detection in PBMC.Methods One hundred and eight patients with chronic HBV infection were involved in this study.PBMC were extracted using density gradient centrifugation.HBV cccDNA in PBMC and serum HBV DNA were detected by real-time fluorescence quantitative polymerase chain reaction.HBsAg and HBeAg were detected by chemiluminescence immunoassay.Liver biopsy was conducted in 59 out of the 108 patients.Chi-square test was used to compare the categorical variables.Correlation analysis was used to compare quantitative variables.Nonparametric test was used to compare the non-normal distribution parameters.Results In the overall population,HBV cccDNA in PBMC was positive in 59 patients (54.6％).Eleven of the 15 patients with liver failure were found to be HBV cccDNA positive,which was significantly higher than that in the acute hepatitis B group (only 2 of the 8 patients were HBV cccDNA positive; x2 =4.960,P＜0.05).One hundred and eight patients were categorized into three groups according to their serum HBV DNA levels,with group A:＞5 lg copy/mL,group B:3-5 lg copy/mL and group C:＜3 lg copy/mL.The proportions of HBV cccDNA positivity in PBMC in three groups were 76.1％ (51/67),5/18 and 13.0％ (3/23),respectively.Comparing with patients with lower HBV DNA (group B and C),the proportion of HBV cccDNA positivity was higher in patients with higher HBV DNA (group A; x2=14.751,P＜0.05 and x2 =28.384,P＜0.05,resepectively).The HBV cccDNA quantitation in PBMC was positively correlated with the serum HBV DNA level and HBsAg quantification (r=0.554,P＜0.05 and r=0.497,P＜0.05,respectively).The proportion of HBV cccDNA positivity in PBMC of patients with liver histology ≥G2 and/or ≥S2 was significantly higher than that in patients with liver histology ＜ G2/S2 (x2 =9.159,P＜0.05).Conclusions HBV cccDNA exists in PBMC of hepatitis B patients.The HBV cccDNA quantitation in PBMC is positively correlated with the serum level of HBV DNA and HBsAg quantification,and is also associated with liver histology injury.

AIM:To investigate the effect of ovarian carcinoma cells on ? chain expression and the secretion of Tc1/Tc2 type cytokine in CD8+ T cells,and its role in the ovarian carcinoma induced immunosuppression.METHODS:The supernatants of human ovarian carcinoma cell lines of OVCAR3,CAOV3 and SKOV3 and RPMI-1640 were added into CD8+ T cells(groups I,II,III,and control),which were isolated from the peripheral venous blood of healthy persons.The expression of ? chain was analyzed by Western blotting.Thiazolyl blue(MTT)method was used to detect the effects of those cell line supernatants on the growth of CD8+ T cells.The secretion of the Tc1 type cytokine interferon(IFN)-? mRNA and the Tc2 type cytokine interferon(IL)-10 mRNA were detected by RT-PCR.RESULTS:The expression of ? chain was significantly lower in groups I,II,and III in comparison with that in control group.The absorbance at the wavelength 570 nm of CD8+ T cells culture in the group I,II,and III was all significantly lower than that in the control group.The IFN-? expression was significantly lower in groups I,II,and III in comparison with that in control group,while the expression of IL-10 was significantly higher.CONCLUSION:Ovarian carcinoma may suppress CD8+ T cell proliferation and secretion of the Tc1/Tc2 type cytokine through inhibition of ? chain,which may play an important role in the ovarian carcinoma induced immunosuppression.

Objective　To ascertain the natural infection rate of hemorrhagic fever with renal syndrome virus (HFRSV) among its hosts and the type of the natural foci for providing some baseline data for the immigrant health and epidemic prevention of the Three-Gorge region. Methods　Epidemiological survey on the field was performed including epidemiological data collection, ecology of rodents and pathogen detection. HFRS virus antigen of hosts were detected by the direct immunofluorescent assay (DIFA) technique and determination of HFRSV-RNA by ISH were carried out from HFRSV-Ag-positive animals. Results　HFRSV-Ag-positive animals were found in 5 migration areas ie Baitao Town of Fuling Section, Wansheng Village of Fengjie County and Dachang Town of Wushan County. The positive hosts were as follows, Rattus Norvegicus, Apodemus agrarius, Anourusurex squamipes, Mus musculus and Rattus flavipectus. The positive rate of HFRSV in the mice of 5 migration areas were 19.4%, 17.0%, 14.0%, 13.7%, and 8.5% respectively. The results showed that the lung tissues of some hosts in all five migration areas were HFRSV-RNA-positive. Baitao Town and Peishi Town were attributed to mixture type epidemic areas while. Kangle Town, Wansheng Village and Dachang Town were domestic rats type epidemic areas. Conclusion　This study shows that the five migration areas are natural epidemic foci of HFRS. It is predicted that maximum risk of HFRS breakout or epidemic may take place after the completion of the San Xia Reservoir(the Three-Gorges Reservoir), which results from rodent moving toward higher land. Therefore, deratization and preventive measures for rat are important in migration areas.

OBJECTIVETo investigate the correlation between intrahepatic eovalently closed circular (ccc)DNA of hepatitis B virus (HBV) and pathogen-and patient-related parameters.

METHODSUltrasound-guided liver biopsies were obtained from 60 patients with chronic HBV infection. Levels of intrahepatic HBV cccDNA and serum HBV DNA were measured by quantitative fluorescence PCR. Level of serum hepatitis B surface antigen (HBsAg) was measured by chemiluminescence immunoassay. Clinical parameters, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (AKP), albumin, globulin (GLO), white blood cell, platelet, prothrombin-international normalized ratio, were measured by standard assay. Demographic information was recorded.The correlation between intrahepatic HBV cccDNA and pathogen-and patient-related parameters was assessed.

RESULTSIntrahepatic HBV cccDNA level was negatively correlated with age, GLO, ALT and grade of necroinflammation. Patients with age of 30 years or more showed significantly higher level of HBV cccDNA level than patients under 30 years-old (7.44±0.58 and 5.66±1.35; t=7.157, P less than 0.001). Intrahepatic HBV cccDNA level was positively correlated with serum HBV DNA level (r=0.916, P less than 0.001) and serum HBsAg level (r=0.727, P less than 0.001). The median ratio of HBV cccDNA to HBV DNA was 1.18, and of HBV cccDNA to HBsAg was 1.67.

CONCLUSIONIntrahepatic HBV cccDNA levels decrease with age, level of ALT, level of GLO and grade of liver necroinflammation, but increase with level of serum HBV DNA and level of serum HBsAg. To a certain extent, serum HBV DNA and serum HBsAg levels may be a sufficient marker of intrahepatic HBV cccDNA levels.

Age Distribution ; Alanine Transaminase ; Aspartate Aminotransferases ; Biomarkers ; DNA, Circular ; DNA, Viral ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Real-Time Polymerase Chain Reaction ; Serologic Tests

Objective To evaluate the impact of Montessori-based intervention on eating ability of the elderly with dementia.Methods Sixty-four patients with dementia were randomly divided into intervention group(n=32) and control group(n=32).Patients in intervention group were given Montessori-based intervention,and patients in control group received regular care.The effect of Montessori-based intervention on eating ability,eating difficulty and self-eating time were assessed by the simplified Chinese version of EBS (C-EBS),simplified Chinese version of Edinburgh feeding evaluation in dementia scale (C-EdFED) and stopwatch respectively at pre-intervention,post-intervention,1-month and 3-month follow-up.Results Compared with the control group (EBS:(12.42± 3.59);EdFED:(10.48± 3.83);self-eating time:(15.28±6.04) min)),the average scores of EBS(14.31±2.63) increased and the self-eating time ((21.44±7.17)min) increased after 8 weeks intervention in intervention group,while the average scores of EdFED (7.86±4.16) increased.The C-EBS scores and self-eating time in intervention group were significantly higher than that of control group while the C-EdFED scores were lower than that of control at all time points(P＜0.01).The difference in time effect between the two groups was statistically significant (P ＜ 0.05)Conclusion The Montessori-based intervention can improve the eating ability of elderly people with dementia,reduce eating difficulty and increase self-eating time.

Objective:To observe the effect of feedback pulmonary rehabilitation guidance on self-management of elderly patients with stable chronic obstructive pulmonary disease (COPD).Methods:Ninety-four elderly patients with stable COPD that visited the outpatient department between January 2018 and January 2019 were selected. Block randomization methods were used to divide the patients into two groups: routine lung rehabilitation instruction group (referred to as the “routine group”) and feedback lung rehabilitation group (referred to as the “feedback group”). The feedback group received the instruction of feedback pulmonary rehabilitation guidance, while the routine group received the instruction of routine pulmonary rehabilitation guidance. The quality of life and the self-management ability of the two groups before and after the intervention were compared.Results:There was no statistically significant difference between the scores of the feedback and routine groups before the intervention of the COPD-specific self-management scale ( P>0.05). In the feedback group, the scores of emotional management, daily life management, symptom management, self-efficacy management, and information management on the COPD-specific self-management scale after the intervention were 45.01±5.31, 53.10±6.60, 25.88±3.03, 35.01±5.31, and 24.32±4.20, respectively, whereas those for the routine group were 40.23±5.19, 48.02±6.58, 22.88±3.01, 31.01±4.80, and 20.30±2.88, respectively. The scores of the feedback group were higher than the routine group and the difference was significant ( P<0.001). There was no significant difference between the feedback group and the routine group before the intervention of the respiratory disease questionnaire, i.e., the airways questionnaire 20-revised (AQ20-R) score ( P>0.05). The AQ20-R score of the feedback group after the intervention was 7.22±1.08, which was lower than that of the routine group (9.01±2.01); the difference was significant ( P<0.001). Conclusion:The application of feedback pulmonary rehabilitation guidance in the self-management of elderly patients with stable COPD can improve both their self-management ability and quality of life.

This paper was aimed to study the effect of Qing-Chang Wen-Zhong (QCWZ) decoction on interferon gamma induced protein 10 (IP10) in colon tissues of rats with ulcerative colitis (UC).The UC model was induced using 4.5％ DSS added to distilled water for 7 days.At the same time,low-,medium-and high-dose of QCWZ decoction and mesalazine was given by gavage route daily.Then,the rats were killed and the colon tissues were taken.Expression level of interleukin-1 alpha (IL-1α),IL-1β,IL-6,tumor necrosis factor alpha (TNF-α) and interferon gamma (INF-γ) in colon were detected by Elisa assay.The expression and distribution of IP10 protein were detected by immunohistochemistry (IHC).The results showed that compared with the normal group,inflammatory factors (IL-1α,IL-1β,IL-6,TNF-α,INF-γ) and IP10 expression level in DSS-induced UC rats were significantly increased.After 7 days of intervention,inflammatory factors (IL-1α,IL-1β,IL-6,TNF-α,INF-γ) and IP10 decreased significantly (p＜0.01,p＜0.05).It was concluded that QCWZ decoction may down-regulate the expression of IP 10 and inflammatory factors (IL-1α,IL-1β,IL-6,TNF-α,INF-γ),and then inhibit intestinal inflammation and repair intestinal mucosal damage,so as to achieve the purpose of UC treatment.