OBJECTIVETo explore the relationship between the temperature and time in moxa cone moxibustion with different purities of moxa.

METHODSAccording to the purities, the moxa were divided into a 10 : 1 group, a 20 : 1 group, a 30 : 1 group and a 40 : 1 group, 30 moxa cones in each group. With the VICTOR DM6902 electronic thermometer, the temperature of the undersurface center at different time points during the moxa cone moxibustion with different purities of moxa was measured. Once the moxa cone was ignited, the results were recorded at the end of each second. Also the temperature peak of the undersurface center and the time when the peak occurred were recorded.

RESULTThe undersurface temperature was increased in all the groups; the time of moxa cone reaching the lowest peak temperature was significantly different in all the groups (all P<0. 05), which was the 10 : 1 group, 20 : 1 group, 30 : 1 group and 40 : 1 group from slow to fast. 50 s, 60 s, 70 s, 80 s and 90 s after moxa cone was ignited, the temperature of moxa cone at the same time point was significantly different in all the groups (all P<0. 05), which was the 10 : 1 group, 20 : 1 group, 30 : 1 group and 40 : 1 group from slow to fast. Conclusion Among the moxa with purity of 10 : 1, 20 : 1, 30 : 1, and 40 : 1, the temperature change rate of the low-purity moxa cone is smaller than that of higher purity, and the stimulating duration of the former is longer than the latter. It is believed that the moxa with purity of 40 : 1 is suitable for scarring moxibustion; the moxa with purity of 30 : 1 and 20 : 1 is suitable for the non-scarring moxibustion; the moxa with purity of 10 : 1 is suitable for gentle moxibustion therapy. The high-purity moxa can also be applied to the field of the low-purity moxa.

Acupuncture Points ; Artemisia ; chemistry ; Humans ; Moxibustion ; instrumentation ; methods ; Plant Leaves ; chemistry ; Temperature ; Time Factors

Objective To develop a time saving and sensitive cell culture system based on hepatitis C virus chimera expressing enhanced green fluorescent protein(EGFP) and to facilitate the study on HCV pathogenesis and screening of anti-HCV drugs.Methods Enhanced green fluorescent protein reporter gene and a mutation V2440L that can yield higher virus titers were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Viral RNA in supernatant of HCV RNA-transfected cells was determined after transfection by RT-PCR.HCV replication and infection were determined by immunofluorescence assay.IFN-α was used to evaluate the feasibility of this system for anti-HCV drugs screening.Results The viral RNA replicated efficiently in transfected cells.These cells can produce HCV-EGFP reporter virus.Viral RNA levels in supernatant were 3.06× 105 copies/ml and 7.96×106 copies/ml at 72 h and 9 d after transfection,respectively.The virus titer reached to 104 FFU/ml 9 d after transfection.The expression of EGFP was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-EGFP reporter virus.Conclusion The recombinant HCV JFH1-EGFP reporter gene system is a time saving,cost effective and sensitive method for studying viral replication cycles and screening of anti-HCV drugs.

ObjectiveTo develop a cell culture system with consistent expression of whole hepatitis C virus (HCV) gene and Renilla luciferase gene and to facilitate the study on HCV pathogenesis and the screening of new antiviral drugs.MethodsRenilla luciferase (RLuc) reporter gene and a mutation that could yield higher virus gene expression were introduced into the C-terminus of non-structural protein 5A (NS5A) of the JFH1 viral genome by using recombinant PCR.The viral RNA was transfected into Huh7.5 cells.Naǐve Huh7.5 cells were infected by the supernatant from the viral RNA transfected cells.HCV replication and infection were determined by virus titration,Renilla luciferase assay,immunofluorescence assay and western blotting.IFN-α was used to evaluate the feasibility of this system for anti-HCV new drug screening.ResultsThe viral RNA replicated efficiently in transfected cells.These cells could produce high titer of HCV-Rluc reporter virus and the virus titer reached to 1.5 × 104 FFU/ml at day 15 of posttransfection.The activity of Renilla luciferase was inhibited by IFN-α in a dose dependent manner in Huh7.5 cells infected by HCV-Rluc reporter virus.ConclusionThe recombinant HCV-JFH1-Rluc reporter gene system is sensitive and efficient.It can be a useful tool for high throughput screening of anti-HCV drugs.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate both mRNA and protein expression of target genes and play important roles in proliferation,differentiation,development and metabolism of cells.This paper reviews the research progress on miRNAs involvement in liver diseases,including viral hepatitis,fatty liver,drug induced liver disease,primary biliary cirrhosis and primary hepatocellular carcinoma.

Objective To investigate the expression of microRNA-548ah (miR-548ah) and microRNA-4804 (miR-4804) in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis B virus infection and their clinical significance.Methods PBMCs were collected from 29 patients with chronic hepatitis B (CHB),30 hepatitis B virus carriers (HBVC),26 inactive HBsAg carriers (IASC) and 28 healthy controls in Taizhou People's Hospital during September 2012 and August 2013.Expressions of miR-548ah and miR-4804 in PBMCs were detected by fluorescence real-time quantitative RT-PCR (qRT-PCR).Receiver operating characteristic (ROC) curve was used to evaluate the expression of miR-548ah and miR-4804 in distinguishing immune tolerance phase and clearance phase of HBV infection.Pearson correlation analysis was performed to assess the correlations of miRNAs expression with clinical markers alanine aminotrans ferase (ALT) and HBV DNA loads.Results There were significant differences in expressions of miR-548ah and miR-4804 in PBMCs between CHB,HBVC,IASC groups and control group (F =28.16 and 83.17,P ＜ 0.01).Compared with control group,miR-548ah was up-regulated in CHB group(P ＜ 0.01),and down-regulated in HBVC and IASC groups (P ＜ 0.01) ; miR-4804 was up-regulated in CHB group (P ＜ 0.01),down-regulated in HBVC group (P ＜ 0.01),while there was no significant difference between IASC group and control group in miR-4804 expressions(P ＞ 0.05).The areas under ROC curve (AUCs) of miR-548ah and miR4804 in differentiation of immune tolerance and immune clearance were 0.966 and 0.997,and the sensitivities and specificities were 89.7％,96.6％ and 99.6％,100.0％,respectively.No significant correlation was found between the expression of miR-548ah,miR-4804 and ALT,HBV DNA loads (r=0.14,0.18,-0.20 and-0.19,P＞0.05).Conclusion miR-548ah and miR-4804 may be involved in the immunopathogenesis of CHB,and their expression levels in PBMCs are helpful in differentiation of immune tolerance and immune clearance in HBV infection.

The sterile stem tips, nodular and non-nodular stem of Morinda officinalis How were cultured in vitro. The results showed that the most suitable procedure for sterilization of explants was soaking into 0.1% HgCl 2 solution for 10 to 15 minutes after pretreating with 70% ethanol for 30 seconds. MT culture medium with BA was effective to induce direct shooting of stem tips and nodular stem, the shooting rate of nodular stem being 97.5% . The optimal BA concentration was 1 mg?L -1 , the shooting rate would decrease when the concentration of BA increased. As for the induction of rooting, MT culture medium adding with 0.2 to 0.5 mg?L -1 NAA was optimal, the rooting rate being over 80.0% .

Traditionally regarded as a vitamin regulating calcium and phosphorus homeostasis, vitamin D is now discovered as a highly versatile molecule involved in immunity, cancer, infectious diseases, fibrosis, fatty liver diseases, and alcoholic liver diseases.In several studies, lower vitamin D status has been found to be associated with increased risk and unfavorable outcome of acute infections.This paper reviews the research progress of the roles played by vitamin D in various inflammatory diseases and its mechanisms.

Objective To compare the efficacy and safety of tenofovir (TDF) and entecavir (ETV) in patients with chronic hepatitis B (CHB) .Methods Studies that compared the treatment efficacy and safety between TDF and ETV in CHB patients were searched through electronic databases before Mar 2015 .Alanine aminotransferase (ALT) normalization rate ,hepatitis B virus (HBV) DNA suppression rate ,hepatitis B e antigen (HBeAg ) seroconversion rate ,drug resistance rate and safety profile were reviewed .RevMan 5 .2 was used for analysis .Results A total of 13 studies met inclusion criteria and 1 934 patients were analyzed ,including 884 patients treated with TDF and 1 050 with ETV .The HBV DNA suppression rate of TDF was superior to ETV at week 48 (OR=1 .36 ,95% CI:1 .05 -1 .76 ,P=0 .02) .The ALT normalization rate of ETV was superior to TDF at week 24 (OR= 0 .68 ,95% CI:0 .48-0 .96 ,P= 0 .03) .The virological response at week 24 ,ALT normalization rate at week 48 and serological response at week 24 and 48 were not significantly different between patients treated with TDF and ETV (all P>0 .05) .The resistance rate was not significantly different between patients treated with TDF and ETV 24 months after treatment (P=0 .51) .And the safety profiles of these two drugs were similar (P>0 .05) .Conclusions TDF has better virological response compared with ETV .The drug resistance rate and safety profile are similar between TDF and ETV .

ObjectiveTo explore the clinical significance of serum hepatitis B virus large surface protein (LHB), HBsAg, hepatitis B virus (HBV) DNA, and alanine aminotransferase (ALT) levels and their correlation with liver inflammation activity in patients with chronic hepatitis B (CHB). MethodsOne hundred and two HBeAg-positive patients with CHB who visited the People′s Hospital of Taizhou from January 2011 to December 2014 were enrolled in the study, and were divided into mild inflammation group (G0-1) and severe inflammation group (G2-4) according to the grade of liver inflammation activity. Continuous data were expressed as x±s; the t-test was applied for comparison between two groups, and analysis of variance was applied for comparison between multiple groups. The Spearman rank correlation test was applied to investigate the correlation between LHB, HBsAg, HBV DNA, and ALT levels and the grade of liver inflammation activity. The area under the receiver operator characteristic (ROC) curve (AUC) was applied to evaluate their diagnostic values. ResultsThe level of LHB tended to increase with the increasing HBV DNA. The AUCs of LHB, HBsAg, ALT, and HBV DNA for diagnosing severe liver inflammation were 0.763, 0.756, 0.702, and 0.581, respectively, and the diagnostic efficiency of LHB, HBsAg, and ALT reached a moderate level (AUC of the ROC curve 0.70-0.90); the diagnostic values of serum LHB and HBsAg levels for severe liver inflammation were higher than that of serum ALT level. The optimal cut-off for serum LHB level to diagnose severe inflammation was 24.6, and the corresponding sensitivity and specificity were 73.4% and 60.3%, respectively. ConclusionSerum LHB and HBsAg levels have certain predictive values for severe liver inflammation in HBeAg-positive patients with CHB.

Clinical data of 113 patients with non-alcoholic fat liver disease (NAFLD) diagnosed by liver biopsy from January 2015 to January 2017 in Taizhou People's Hospital were retrospectively reviewed . Patients all underwent transient elastographic ( TE) examination and the values of fat attenuation index (FAI) were obtained.The hepatocyt fatty changes in pathological examination were scored as 0 (＜5%, n＝40), 1 (5%-33%,n ＝27), 2 (34% -66%,n ＝28) and 3 (＞66%, n ＝18).There were significant differences in AST , Glu, TC and FAI among patients with hepatocyte fatty change scores 0, 1, 2 and 3, and the FAI was significantly correlated with the degree of fatty liver disease .The areas under the ROC curve (AUCs) of FAI in patients with hepatocyte fatty change scores 1, 2 and 3 were 0.78, 0.90 and 0.96, respectively.Logistic regression analysis showed that FAI was correlated with TG , TC and BMI.The results suggest that FAI in TE can be a non-invasive, rapid and objective evaluation method for patients with NAFLD.