In order to find microRNA associated with HBV infection and to explore the mechanism of the infection, first of all, we found in our preliminary study that in HepG2 cells transfected with HBV expression plasmid, miR-122 expression was up-regulated, suggesting that miR-122 was related to the HBV infection. On this basis, in the present study, miR-122 and pCH9-HBV1.1 plasmid were cotransfected into HepG2 cells. Southern blot detection result showed that miR-122 can inhibit HBV replication. Using MiRanda computer software, HBx was predicted to be the target sequence of miR-122; Luciferase reporter gene system and Western blot detection of HBx protein expression changes were further used to verify the HBx expression regulated by miR-122. And finally, it can be speculated that miR-122 may affected HBV replication by regulating the expression of HBx.
Gene Expression Regulation, Viral ; drug effects ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; genetics ; physiology ; Humans ; MicroRNAs ; genetics ; Trans-Activators ; genetics ; metabolism ; Transfection ; Virus Replication

Objective To develop a vulnerable plaque targeting ultrasound contrast agent (UCA) and to evaluate its affinity and imaging performance in vitro.Methods E-selectin receptor-targeting UCA,which conjugated with monoclonal antibody of E-selectin,was prepared with filming-rehydration method and biotin-avidin linkage.The size and distribution of UCA were measured with particle size analyzer,the connectivity condition of microbubbles with E-selectin antibody was also detected with fluorescence analysis.The cytotoxicity from microbubble and ultrasound irradiation was evaluated through cell counting kit-8 (CCK8) assay.The adhesion effect of UCA was assessed after co-incubated with activated mouse endothelial cells (bEnd.3) and compared with that of free antibody intervention group and control group.The imaging performance of UCA at different time points was observed on an ultrasound equipment with a high-frequency transducer.Two-sample t test and one-way analysis of variance were performed to analyze the data.Results E-selectin receptor-targeting UCA was successfully prepared.The cytotoxicity result with CCK8 assay demonstrated the favorable biocompatibility of UCA.The connection amount of UCA on activated bEnd.3 cells ((6.23 ± 0.45) bubbles/cell) was significantly higher than that of the free antibody intervention group ((1.57±0.34) bubbles/cell) and control group ((0.07±0.03) bubbles/cell;F=291.43,P＜0.01).The performance of in vitro ultrasonography at the same time points showed no obvious difference between targeting UCA and control UCA (all t＜0.51,all P＞0.05).Conclusions The prepared E-selectin receptor-targeting UCA has favorable targeting and imaging capabilities.It might be a potentially ultrasound molecular imaging agent for early detection and prognosis evaluation of vulnerable plaque.

Objective To study the effect of radiofrequency ablation (RFA) with sublethal temperature on the production of liver cancer stem cells (LCSCs) and on the expression of LCSCs-related transcriptional factors.Methods Mouse hepl-6 hepatoma cell line and clinical samples of patients with hepatocellular carcinoma (HCC) were used to test the expressions of LCSCs-related markers and transcriptional factors.Results Different temperatures were used to stimulate Hep1-6 cells,and it was proved that the temperature of 45℃ was a sublethal temperature that could not induce cell death.Flow cytometry testing showed that treatment with 45℃ could obviously increase CD13+,CD44+,CD90 and CD133+ Hep1-6 cells,suggesting that treatment with 45℃ could increase the production of above mentioned types of LCSCs in hep1-6 cells.Real-time quantitative polymerase chain reaction (RT-qPCR) assay indicated that the temperature of 45℃could cause significant increase in CD13,CD90 and CD133 mRNA.In all 5 HCC patients,CD13 mRNA in the recurrent HCC lesions was remarkably increased,CD133 mRNA was increased in 4 patients with recurrent HCC,and CD90 mRNA was increased in only one patient with recurrent HCC.Flow cytometry testing revealed that CD13+ LCSCs were strikingly increased in 4 recurrent HCC patients,while CD133+LCSC was increased in only one patient,suggesting that more close correlation existed between the increase of CD13+ LCSCs and the temperature of 45℃.RT-qPCR assay showed that in 4 recurrent HCC patients with increased CD13+ LCSC,the Sox2 and Stat2 among 13 LCSCs-related transcriptional factors were obviously increased.Flow cytometry testing showed that 45℃ treatment also increased the expression of Sox2 and Stat1 mRNA in Hep1-6 cells.Finally,Sox2 and Stat1 could be knockdown by siRNAs,indicating that both Sox2 and Stat1 transcriptional factors were involved in 45℃-induced production of CD13+ LCSCs in Hep1-6 cells.Conclusion In RFA therapy,the use of sublethal temperature of 45℃ can increase CD13+LCSCs,which is related to the promotion of Sox2 and Stat1 expression.The results of this study can be used for reference in the research of liver cancer recurrence.

Objective:To investigate the prevalence of lymphocytic choriomeningitis virus (LCMV) and hantavirus (HV) specific antibodies among cross-border migrant workers for assessment of the risk of rodents-borne virus infection.Methods:From 2019 to 2020, a survey was conducted on cross border migrant workers engaged in outdoor activities, and serum samples were collected, LCMV specific IgG antibody was detected by an indirect ELISA and Western blot based on recombinant nucleoprotein, and indirect immunofluorescence assay (IFA) based on recombinant expressed glycoprotein. HV IgG antibody in serum was detected by a commercial indirect IgG ELISA kit and IFA based on hantavirus infected Vero cells.Results:A total of 139 cross-border workers, aged 25~57, were surveyed; 64% (89/139) had working experience in multiple countries, involving 26 countries, including 14 countries in Asia and 12 countries in Africa; 11.51% (16/139) of serum samples were tested positive for LCMV antibodies, and the positive samples were verified by Western blot and IFA. The antibody detection rate was slightly higher than the published infection rate from other similar studies. And, HV antibodies were detected from one serum sample (0.72%, 1/139) by ELISA and IFA. However, it was still uncertain when and where the viral infections were acquired.Conclusions:Through this serological cross-sectional preliminary analysis, the infection status and existing risks of LCMV and HV viruses among cross border migrant workers were revealed, which suggested the necessity of strengthening the prevention and control of rodents borne diseases in outdoor engineering sites.

OBJECTIVE：To systematically evaluate therapeutic efficacy of Gongliuqing capsules combined with mifepristone in the treatment of uterine leiomyoma ，in order to provide evidence-based reference for clinical medication. METHODS ：Retrieved from Cochrane Library ，PubMed，Embase，CJFD，VIP，CBM and Wanfang database ，randomized controlled trials （RCTs）about Gongliuqing capsules combined with mifepristone （trial group ）versus mifepristone alone （control group ）in the treatment of uterine leiomyoma were collected. After literature screening and data extraction ，the quality of included literatures was evaluated with modified Jadad scale. Meta-analysis was conducted by using Stata 14.0 software，and trial sequential analysis （TSA）was performed by using TSA 0.9 software. RESULTS ：A total of 12 RCTs were included ，involving 1 210 patients. The results of Meta- analysis showed that the total response rate of trial group [RR ＝1.12，95％CI（1.00，1.26），P＜0.05] was significantly higher than that of control group ；maximum uterine leiomyoma volume after treatment [SMD ＝－1.08，95％CI（－1.21，－0.95），P＜0.05]，uterine volume after treatment [SMD ＝－0.80，95％CI（－1.14，－0.45）， P＜0.05]，follicle stimulating hormone （FSH）level [SMD ＝ － 0.28，95％ CI（－ 0.45，－ 0.19），P＜0.05]，luteinizing hormone（LH）level [SMD ＝－0.44，95％CI（－0.52，－0.12）， 020-38076311。E-mail：867203217@qq.com P＜0.05]，E2 level [SMD ＝－2.69，95％CI（－3.08，－1.49）， P＜0.05] and progesterone （P）level [SMD ＝－1.27，95％CI（－1.69，－0.71），P＜0.05] of trial group were significantly lower or better than those of control group. Results of subgroup analysis showed that except for the level of FSH in 5 and 10 mg mifepristone groups （P＞0.05），maximum uterine leiomyoma volume after treatment ，uterine volume after treatment ，the levels of FSH，LH，E2 and P in trial group were significantly lower than control group. The results of TSA showed that there were definite evidences for total response rate of Gongliuqing capsules combined with mifepristone being better in the treatment of hysteromyoma. CONCLUSIONS ：Total response rate of Gongliuqing capsules combined with mifepristone in the treatment of hysteromyoma is better than mifepristone alone ，which can effectively decrease the volume of maximum uterine leiomyoma volume and uterine vilume ，and reduce the level of serum hormone.