Objective To evaluate the inhibitory effect of the small interfering RNA(siRNA) on hepatitis B virus(HBV)in vivo which targets HBV S gene region.Methods An animal model of HBV infection was developed hydrodynamically by injecting pcDNA3.1-HBV together with siRNA through the tail vein of Balb/c.HBsAg was analyzed by time resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR(FQ-PCR),HBV S-mRNA was detected by semi-quantitative RT-PCR,and viral specific proteins(HBsAg and HBcAg)in the liver were assayed by immunohistochemical staining.Results In the mice,the siRNA could effectively inhibit the secre- tion of HBsAg,reduce the titers of HBV DNA,and immunohistochemical results also indicated that the number of HBsAg and HBcAg positive cells was reduced.The inhibitory effect of siRNA on HBV lasted 3 clays at least.Conclusion These results demonstrate that the siRNA targeting HBV S gene region can substantially and specifically inhibit HBV replication and expression in vivo.

Objective To investigate the effect of sodium fluoride(NaF) on proliferation, differentiation and the mRNA expression of osteoprotegerin (OPG) and receptor activator of nuclear factor κβ ligand (RAN KL) of mouse osteoblasts. Methods Osteoblasts were isolated from calvarias of Kunming mice born in 1 - 2 d and cultured. Various concentrations of NaF(0, 10-8, 10-7, 10-6, 10-5, 10-4, 10-3mol/L) were added to the culture medium, the proliferation and activity of alkaline phosphatase(ALP) was determined after 72 h or 120 h. The expression of OPG mRNA and RANKL mRNA was analyzed by semi-quantification RT-PCR. Difference among groups was analyzed by One-Way AN0VA. Difference between two groups was analyzed by LSD-t test. Results There was significant difference in cell proliferation among groups after 72 h(F = 13.806, P < 0.05). Compared with control group(0.434 ± 0.010) , the proliferation was significantly induced in 10-7 - 10-4 mol/L groups treated osteoblasts (0.448 ± 0.010, 0.453 ± 0.013, 0.454 ± 0.016, 0.449 ± 0.018, all P< 0.05), and was significantly suppressed in 10-3 mol/L group(0.401 ± 0.009, P < 0.05). There was statistic difference in the activity of ALP among groups(F = 9.021, P < 0.05). Compared with control group (1.677 ± 0.682), the activity of ALP significantly increased in 10-7 - 10-5 mol/L groups[ (2.447 ± 0.756) × 106, (2603 ± 0.183) × 106, (2.687 ± 0.886) × 106 U/L, P < 0.05 or P < 0.01 ] and significantly decreased in 10-4 mol/L group[ (1.479 ± 0.366) × 106 U/L, P < 0.05 ]. There was significant difference in the expression of OPG mRNA among groups(F = 11.299, P< 0.05). Compared with control group (1.000 ± 0.000), the expression of OPG mRNA was significantly increased in 10-7 - 10-4 mol/L groups( 1.058 ± 0.027, 1.053 ± 0.026, 1.088 ± 0.055, 1.069 ± 0.008, P < 0.05 or P < 0.01) , while significantly decreased in 10-3 mol/L group (0.941 ± 0.029, P< 0.05). There was no difference in RANKL mRNA expression among groups (F= 1.311, P> 0.05). The ratio of RANKL/OPG decreased with increasing doses of fluoride and increased in 10-4, 10-3 mol/L groups, but there was no difference between groups(F = 1.376, P> 0.05). Conclusions A biphasic pattern of proliferation and differentiation has been induced in mouse osteoblasts, which manifests stimulation effect in low doses and suppression in higher doses. Low doses of sodium fluoride suppress differentiation and maturation of osteoblasts by increasing expression of OPG mRNA, while high doses of sodium fluoride enhance differentiation and maturation of osteoblasts by decreasing expression of OPG mRNA.

Objective To observe the dynamic changes of NO,NOS in serum and CSF in patients with CNS infection and lay an experimental basis for discrimination of CNS infection due to different agents.Methods The method of nitrate redutase and the method of chromometry were employed to measure NO,NOS in serum and CSF at different time.The dynamic changes of NO,NOS in serum were observed on admission,on 3,5,9,14 day after admission.The dynamic changes of NO,NOS in CSF before treatment and two weeks after treatment were ob- served,too.Results There were no difference between the the concentration of NO and the vigor of total NOS in serum and in CSF of viral meningitis,bacterial meningitis and tubercular meningitis patients due to different agents. Conclusion The changes of the concentration of NO in serum and CSF,the vigor of total NOS in serum and CSF could not he seen as laboratory basis for discrimination of CNS infection due to different agents.

OBJECTIVETo evaluate the inhibitory effect of small interfering RNA (siRNA) targeting HBV C gene region on hepatitis B virus (HBV) in vivo.

METHODSAn animal model of HBV infection was developed hydrodynamically, and pcDNA3.1-HBV and siRNA were together injected into the tail vein of the BALB/c mice. HBsAg was analyzed by time-resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR, and viral specific proteins (HBsAg and HBcAg) in the mice livers were assayed using immunohistochemical staining.

RESULTSIn the mice, the siRNA effectively inhibited HBV replication and expression compared with the controls. The inhibitive effect of siRNA on HBV lasted at least 3 days.

CONCLUSIONThese results demonstrate that RNAi can substantially inhibit HBV replication and expression in vivo.

Animals ; Female ; Hepatitis B ; therapy ; Hepatitis B virus ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; physiology ; RNA-Induced Silencing Complex ; Random Allocation ; Virus Replication ; genetics

OBJECTIVETo design pSilencer3.1-H1hygro plasmid expressing short interfering RNAs (siRNA) that targets HBV core gene region, and to evaluate inhibitory effect of this siRNA on HBV in vitro.

METHODSHepG2 2.2.15 was used as target cells. The plasmid and liposome metafectene were cotransfected into the cultured cells, HBV DNA were analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR.

RESULTSThe plasmid expressing siRNA was successfully constructed. The two constructed siRNAs could effectively inhibit HBV replication, and their inhibitive effect on HBV was dose-dependent.

CONCLUSIONThese results showed that siRNA could substantially inhibit HBV replication in the infected cells

Hepatitis B virus ; genetics ; physiology ; Humans ; Liver Neoplasms ; virology ; RNA Interference ; Tumor Cells, Cultured ; Virus Replication ; genetics

Objective To investigate the prevalence and determinants of depressive symptoms among ‘empty-nest' and ‘non-empty-nest' elderly in four cities/provinces.Methods 4265 elderly aged 60 and over,were recruited with cluster sampling method in Shanghai,Heilongjiang,Guangdong and Shanxi province and interviewed,using the Geriatric Mental State Schedule and self-developed related questionnaire.Results ( 1 ) The prevalence of depressive symptoms for ‘empty-nest' elderly was (8.18％),significantly higher than that for ‘non-empty-nest' eldcrly (P=0.019) ; (2) the ‘empty-nest' elderly had a significantly higher proportion of the following factors:being male,married,with higher income ( ≥ 15 000 Yuan/year),living in city,with high education background,under employment etc.than the ‘non-empty-nest' elderly (P＜0.0001) ; (3)the ‘empty-nest' elderly had significantly higher proportions on good self-rated health status and life (P=0.0001,P＜0.0001 ) as well as heavier health problems and economic difficulties (P=0.00 1,P=0.002 ) ; (4) there were significantly negative associations between depressive symptoms and the following 10 factors:being female,single,having bad self-rated health and life status,having somatic disease ≥3,with big health problems in the last two years and loss of dearest persons,community engagement and involvement of religious activities.Conclusion The ‘empty-nest' elderly showed higher prevalence of having depressive symptoms than the ‘non-empty-nest' elderly.The ‘empty-nest' elderly had characteristics as being single,female,having adverse event etc.and should be under greater attention for care.

Objective To study the relationship between liver pathological changes and serum HBeAg and HBV DNA in patients with chronic hepatitis B. Methods Liver puncture biopsy for histopathological examinations were performed in 1057 patients with chronic hepatitis B. The quantitative analysis of serum HBV DNA by fluorogenic quantitative PCR and HBeAg by chemoluminescence were also conducted. Results The inflammatory grade and fibrosis stage were higher in HBeAg-negative paients (G4 and S4 were 7.83% and 12.17%respectively) than in HBeAg-positive patients (G4 and S4 were 3.39% and 5.44% respectively). The inflammatory grade and fibrosis stage were higher in HBeAg-positive patients with low-level HBV DNA( G3G4 was 45.64% and S3S4 was 30.20% for HBV DNA 104-105 ), whereas they were higher in HBeAg-negative patients with high-level HBV DNA(G3G4 was 54.55% for HBV DNA 106-107 and S3S4 was 42.85% for HBV DNA 108-109). Conclusion There were some correlation between the liver pathological changes and serum HBeAg and HBV DNA levels in patients with chronic hepatitis B. It is important to perform the liver pathological examination and antiviral therapy as early as possible in patients with HBeAg-negative chronic hepatitis B.

To study the liver histopathological features that are distinctive between chronic hepatitis B virus (HBV) infection patients who have normal serum alanine aminotransferase (ALT)/asparatate aminotransferase (AST) and those with mildly elevated serum ALT/AST. One-hundred-and-thrity-four chronic HBV infection patients with normal serum ALT/AST and 165 chronic HBV infection patients with mildly elevated serum ALT/AST were included in the study. Liver biopsies were performed and used to assess the histological changes by hematoxylin-eosin and reticular fiber staining; mild to severe scoring for inflammation was made as grade G0-G4 and for fibrosis stage as S0-S4. HBV DNA levels were detected by fluorescent quantitative PCR. HBV serological markers were examined by chemiluminescence. The mildly elevated serum ALT/AST group had more male patients than the normal serum ALT/AST group. In the normal serum ALT/AST group, 50.0% (67/134) of the patients had moderate histological changes and only 3.0% (4/134) had severe changes (G3-4 and/or S3-4). In the mildly elevated ALT/AST group, 65.7% (174/265) of patients had moderate histological changes and 16.2% (43/265) had severe changes (G3-4 and/or S3-4). Hepatic inflammation and fibrosis were significantly more severe in the mildly elevated serum ALT/AST group than in the normal ALT/AST group (x2 = 26.386, P less than 0.01; x2 = 15.299, P less than 0.01). In the normal ALT/AST group, the severity of inflammation and fibrosis were positively correlated with age (rs = 0.620, P less than 0.01; rs = 0.347, P less than 0.01). In the mildly elevated ALT/AST group, the severity of inflammation and fibrosis were negatively correlated with age (rs = -0.807, P less than 0.01; rs = -0.557, P less than 0.01). In both groups, the severity of inflammation and fibrosis were negatively correlated with HBV DNA levels (rs = -0.215, P less than 0.01, rs = -0.527, P less than 0.01, rs = -0.951, P less than 0.01; rs = -0.715, P less than 0.01) and were not positively correlated with HBeAg. The majority of the chronic HBV infection patients with normal serum ALT/AST and those with mildly elevated serum ALT/AST had moderate liver pathological changes. All patients with low HBV DNA levels were closely followed-up, regardless of HBeAg-positive status.
Adolescent ; Adult ; Age Factors ; Alanine Transaminase ; blood ; Aspartate Aminotransferases ; blood ; Biopsy, Needle ; Child ; DNA, Viral ; blood ; Fatty Liver ; pathology ; virology ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Humans ; Liver ; pathology ; virology ; Male ; Middle Aged ; Retrospective Studies ; Viral Load ; Young Adult

BACKGROUND: Postoperative anticoagulant therapy after hip and knee arthroplasties has been included in the perioperative management guidelines. However, the application of anticoagulant drugs accompanies with the risk of bleeding. Routine coagulation tests provide limited information about the quality of clots because they identify only the first stage of clotting, while thrombelastography provides a comprehensive assessment of coagulation function. But its practicality remains controversial and the research for bleeding after joint replacement is little reported. OBJECTIVE: To explore the distribution of thrombelastography parameters (time to initial fibrin formation, clotting time, α angle, and maximum amplitude) and to analyze the correlation of the four parameters with postoperative blood loss, thereby providing guidance for improving the safety and effectiveness of anticoagulant therapy. METHODS: Totally 148 patients with detection of thrombelastogram after arthroplasty from August 2015 to March 2017 in Sun Yat-sen Memorial Hospital, Sun Yat-sen University were enrolled, including 76 cases of total hip arthroplasty and 72 cases of total knee arthroplasty. Thrombelastography data were collected on day 1 postoperatively, and the perioperative blood loss was calculated. Structural equation modeling of each group was constructed to investigate the relationship of four parameters and total blood loss. RESULTS AND CONCLUSION: (1) In the structural equation modeling of hip and knee arthroplasties, the root mean square error of approximation was less than 0.08, goodness-of-fit index, adjusted goodness-of-fit index, normed fit index and comparative fit index was all higher than 0.9, and Parsi-mony goodness-of-fit index was less than 2, so the theoretical model was matched with the data. (2) There was a correlation of postoperative hemorrhage with time to initial fibrin formation, clotting time, α angle, and maximum amplitude. (3) That is to say, thrombelastogram can be used as an efficient tool in predicting bleeding after hip and knee arthroplasties. Future study based on this research will further verify the correlation and provide more information for its clinical practice.

Aim To investigate the protective effect of Jujing formula on retina of mice with dry age-related macular degeneration(AMD).Methods The mouse model of dry AMD was induced by intraperitoneal in-jection of sodium iodate,and the prognosis was given to the Jujing formula.Retinal thickness was detected by optical coherence tomography(OCT),the retinal morphological changes were observed by hematoxylin-eosin(HE)staining,and the apoptosis of retinal cells was detected by in situ terminal transferase labeling(TUNEL)staining.Combination of tumor necrosis fac-tor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1 β)in eyeballs and serum,superoxide dis-mutase(SOD),glutathione(GSH)and malondialde-hyde(MDA)were evaluated to assess the protective effects of Jujing formula on retinal injury in mice with dry AMD.Results The results of OCT,HE and TUNEL staining showed that Jujing formula significant-ly improved the retinal injury induced by sodium iodate in mice with dry AMD,increased the retinal thickness(P＜0.05),reduced the apoptosis of retinal cells(P＜0.01),and increased the levels of GSH,IL-6 and SOD activity in eyeballs and serum(P＜0.01).The levels of TNF-α,IL-6,IL-1β and MDA were reduced(P＜0.01).Conclusions Jujing formula has certain therapeutic effects on retinal injury in dry AMD,which may be related to inhibiting inflammatory response and enhancing antioxidant capacity.