Objective:To construct the retroviral vector containing HBx gene so as to investigate the effects of HBx gene in the pathogenesis of HBV-dependent hepatic cell carcinoma(HCC)and further construct animal models of hepatic cell carcinoma.Methods:HBx gene was amplified from the HBx adenovirus Plasmid by polymerase chain reaction(PCR)technique whose primer was designed according to the announced sequences of HBx gene in gene bank,then cut by two endonucleases and directionally cloned into the pseb-hus vector and analyzed by gene sequencing analyzer,the correct constructed vectors were transfected into L02 cell line.At last,HBx protein was detected by Western Blotting.Results:Through PCR,endonuclease cutting and gene sequencing,the target gene was verified into be correctly cloned to retroviral plasmid pSEB-HUS and the high expression of green fluorescence protein(GFP)in L02 cell line was observed under fluorescent microscope.The expression of HBx protein was also observed by Western Blotting.Conclusion:Recombinant retroviral vector expressing HBx protein was successfully constructed.

OBJECTIVE: To investigate the protective effect of berberine on CCl4-induced acute liver injury in mice.METHODS: Kunming mice were included and equally divided into 6 groups: normal group,model group,berberine groups(40,20,10 mg?kg-1).After intragastric administration of corresponding drugs for 5 consecutive days,the mice were intraperitoneally injected with CCl4(10 mL?kg-1) to establish liver injury mice model.Alanine transaminase(ALT) and aspartate transaminase(AST) in serum,activities of superoxidase(SOD) and malondialdehyde(MDA) content in liver homogenate were measured.RESULTS: In Berberine-treated groups compared the model group,serum ALT level and AST activity decreased significantly(P

According to the registered product standards specification of medical devices, combined with the standard reviewing work, the common problems of standards during medical devices registration were analyzed and corresponding suggestions were proposed to standardize the standard of the registered product, accelerate the standardization and promote the industry standardized.
Device Approval ; Equipment and Supplies ; standards

This study aimed to investigate the effects of fusion proteins GnRH-GRP(G3G6)and HSP65-STEAP1(HST1)on dendritic cells(DC)and the sensitization of DCs to B16F10 melanoma. The fusion proteins G3G6 and HST1 were obtained using the previous engineering strains in our laboratory. Group by unsensitized DC(US-DC), the G3G6 fusion protein sensitized DC, the HST1 fusion protein sensitized DC(HST1-DC)and the combined sensitized DC(GH-DC), the mouse bone marrow-derived DCs were sensitized with fusion protein to obtain the fusion protein sensitized DC vaccines. B16F10 melanoma cells were transplanted into C57BL/6J male mice to construct a melanoma model(1×106 cells per mouse), and DC vaccine was injected for treatment. The antitumor efficacy of DC vaccine was explored by in vitro and in vivo experiments. Flow cytometry analysis showed that the fusion protein can effectively stimulate DC into differentiation and maturation; in the animal experiment, the inhibition rate of melanoma treated with G3G6-DC was 35. 75%, that of HST1-DC group and combination group were 34. 03% and 55. 74%. It was initially proved that both G3G6-DC and HST1-DC can effectively inhibit the growth of transplanted tumors of melanoma B16F10 cells in mice, and the combination therapy is superior to the single therapy.

Objective To evaluate the relationship between the risk of breast cancer and abortions among nulliparous women. Methods Searched the data of Cochrane Library and PubMed before June 2014 to identify potentially studies which involved the relationship between the risk of breast cancer and abortions among nulliparous women. Data was extracted by two independent authors from each study. STATA software was used for statistical analysis. Calculated the pooled RR and 95% CI as the assessment of the link between abortions and breast cancer in fixed effects models. Results 13 studies were included. The study showed the RR and 95%CI of the relationship between the risk of breast cancer and abortions was 0. 98[0. 89,1. 08],P>0. 05 in nulliparous women, and the number of abortions was not associated with the risk of breast cancer. The RR and 95%CI of the relationship between the risk of breast cancer and induced abortions or spontaneous abor-tions were 0. 96[0. 88,1. 04],1. 01[0. 88,1. 14], respectively. Conclusion There is no correlation between breast cancer and abortions a-mong the nulliparous women, and the risk of breast cancer would not increas as the number of abortions increase.

Objective To investgate role of TLR2 in the activation, the innate immune and inflammation of human platelets. Methods Human washed platelets were separated from healthy people(n=5) and were stimulated with different concentrations(1μg/ml, 5μg/ml, 10μg/ml) of TLR2 agonistPam3CSK4(a synthetic bacterial lipoproteins). Then the platelet aggregation rate, the expression of CD62p and TLR2 on the platelet surface were measured. Results The platelet aggregation rate were (28.32±5.67)%, (52.56±8.54)% and (76.24±11.23)%, respectively, at concentration of 1μg/ml, 5μg/mland 10μg/ml of Pam3CSK4, more than (12.83±2.43)% at 0μg/ml of it. In addition, the expression of CD62p were (18.45±2.66)%, (22.45±2.04)%, (29.53±4.08)%, respectively at above concentration of Pam3 CSK4, more than (11.20±1.67)% of CD62p at control group(P<0.01). The expression of TLR2 was not significantly increased at a lower concentration of Pam3CSK4(1μg/ml) with (16.85±6.10)% compared with(10.81±3.99)% at the control group. However, it were (21.15±9.90)% and (22.52±9.26)%, respectively, at a higher concentration(5μg/ml, 10μg/ml)more than(10.81±3.99)% at the control group(P<0.05). Conclusion Pam3CSK4 induce aggregation, activation and the up-regulation of TLR2 of platelet by stimulating TLR2 receptor of it. Thereafter, TLR2 play an important role in the innate immune of platelet.

For the existing measurement methods of residual voltage which can't turn the power off at peak voltage exactly and simultaneously display waveforms, a new residual voltage detection method is put forward in this paper. First, the zero point of the power supply is detected with zero cross detection circuit and is inputted to a single-chip microcomputer in the form of pulse signal. Secend, when the zero point delays to the peak voltage, the single-chip microcomputer sends control signal to power off the relay. At last, the waveform of the residual voltage is displayed on a principal computer or oscilloscope. The experimental results show that the device designed in this paper can turn the power off at peak voltage and is able to accurately display the voltage waveform immediately after power off and the standard deviation of the residual voltage is less than 0.2 V at exactly one second and later.
Electric Power Supplies ; Equipment Design ; Microcomputers ; Signal Processing, Computer-Assisted

Objective:To observe the effect of conversion from immediate-release tacrolimus (Tac) to extended-release Tac on kidney function in stable kidney transplant recipients.Methods:83 stable kidney transplant recipients who were converted from immediate-release Tac to extended-release Tac in the second people's hospital of Shanxi province from December 2011 to June 2019 were followed up for 12-36 months, and 83 stable kidney transplant recipients who continued to take immediate-release Tac were selected as control group.The changes of kidney function indexes, Tac trough concentration intra-patient variability (IPV) and compliance, the incidence of rejection and the survival rate of grafts and recipients were observed after the conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients.Results:The conversion time from immediate-release Tac to extended-release Tac was (42.76±30.50)months after transplantation. At 24 months after conversion, the serum creatinine (SCr) was significantly lower than that before conversion ( P=0.013), and the estimated glomerular filtration rate(eGFR)was significantly higher than that before conversion( P=0.005). In the experimental group , the SCr was significantly lower than that of the control group at 36 months after conversion ( P= 0.017), eGFR was significantly higher than that of the control group ( P=0.038). In the experimental group, the score of Immunosuppressant Therapy Barrier Scale (ITBS) was (20.23±2.89) before conversion and (17.63 ±3.08) after conversion ( P= 0.000). The daily dose of Tac was (2.09 ±0.84) mg before conversion and (2.10 ±0.83) mg after conversion. The trough concentration of Tac before conversion was (7.22 ±2.84) ng/mL, which reduced significantly after conversion. No rejection occurred after conversion, and the recipients/grafts survived healthily during the follow-up period. Conclusions:After conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients, the kidney function is stable and better than that of before conversion, the compliance is significantly improved, the IPV of Tac trough concentration is significantly reduced, and long-term use of extended-release Tac has good clinical efficacy and safety.

The immune effect of two recombinant protein fragments of spike protein in severe acute respiratory syndrome coronavirus (SARS CoV) was investigated in Balb/c mice. Two partial spike gene fragments S1 (322 1464 bp) and S2 (2170 2814 bp) of SARS coronavirus were amplified by RT-PCR, and cloned into pET-23a prokaryotic expression vector, then transformed into competent Escherichia E. coli BL21 (DE3)(pLysS) respectively. Recombinant proteins were expressed and purified by Ni2+ immobilized metal ion affinity chromatography. The purified proteins mixed with complete Freund adjuvant were injected into Balb/c mice three times at a two-week interval. High titer antibody was detected in the serum of immunized Balb/c mice, and mice immunized with S1 protein produced high titer IgG1, IgG2a, IgG2b and IgG3, while those immunized with S2 protein produced high titer IgG1, IgG2a, but lower titer IgG2b and IgG3. Serum IFN-concentration was increased significantly but the concentrations of Il-2, IL-4 and IL-10 had no significant change. And a marked increase was observed in the number of spleen CD8+ T cells. The results showed that recombinant proteins of SARS coronavirus spike protein induced hormonal and cellular immune response in Balb/c mice.

ObjectiveTo observe the effect of mesenchymal stem cells (MSCs) on enhancing rat islets viability and function in vitro by a pretransplant co-culture.Methods4-week-old Wistar rats were used as donors, bone mesenchymal stem cells were isolated and subcultured. Islets of Wistar rats were isolated and purified by one-step single-layer Histopaque-1077. Then islets were divided into four groups randomly, 2 groups co-cultured with mesenchymal stem cells (MSCs) (one using low-glucose medium; the other using high-glucose medium ); 2 groups were cultured alone (low-glucose medium; high-glucose medium), each group was further stratified into 3 subgroups(3, 7, 14 d); the survival and functionality of these islets were observed and evaluated. The amount of glucose stimulated secreted insulin were measured wth a rat/mouse insulin enzyme-linked immunosorbent assay (ELISA) kit and stimulation index was also calculated.ResultsCompared with those not co-cultured, islets co-cultured with MSCs demonstrated significantly higher survival rates and viability both in 3th, 7th and 14th day ( P ＜ 0. 01 ); furthermore, cocultured islets revealed higher levels of glucose stimulated insulin secretion and secretion indexes in 7th day (P＜0.01).ConclusionRat islet cells co-cultured with MSCs have longer in vitro survival and better functions.