Objective To determine whether HBx gene can directly induce hepatocellular carcinoma in vivo, and to explore the mechanism of transplantation tumor in nude mice.Methods pCMVX/QSG7701 cell lines were vaccinated into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cell lines were used as controls. The sections of transplantation tumor were observed microscopically by HE staining. RT-PCR was used to detect the expression of mutant p53 and c-Myc mRNA in transplant tumor and an other 3 cell lines. Results The transplant tumor occurred within the subcutaneous tissue of the nude mice inoculated with pCMVX/QSG7701 cell lines at 2nd week after the vaccination. No metastatic tumor was found in other organs. Transplant tumor was not formed in all the controls. HE staining confirmed that the transplant tumor was hepatocellular carcinoma. The mutant p53 mRNA and c-Myc mRNA expression level of transplant tumor and pCMVX/QSG7701 cells was significantly higher than that of pRcCMV2/QSG7701 and QSG7701 cells, respectively (P＜0.01).Conclusion HBx gene can up-regulate the expression of mutant p53 and c-Myc genes, and directly induce hepatocellular carcinoma in vivo.

Objective To investigate gene variation and the relationship between gene variation and pathogenicity of transfusion transmitted virus(TTV).Methods The TTV DNA in the serum sample from a blood donor(BD) and a chronic non-A-G severe hepatitis(CSH) patient with TTV infection was amplified by using PCR.The purified PCR product was cloned and 10 clones from each case were sequenced.The sequences were compared among different clones and analyzed by Phylogentic tree.Results There were two different TTV strains in the BD and seven different TTV strains in the chronic non-A-G severe hepatitis patient.The TTV clones in the BD were of G1a subtype and those of the CSH were of G1a and G1b subtype.Conclusion Gene variant of TTV was much more complicated in the CSH patients than that in the BD ones.

OBJECTIVE:To investigate the anti-HBV activities of deoxynojirimycin(DNJ)derivatives N—benzyl—1—DNJ(P-DNJ)and N—nonyl—1—DNJ(NN-DNJ)in vitro.METHODS:Human hepatoma carcinoma cell lines HepG2 2,2,15,which were transfected from HBV DNA,were taken as target cells,cells were cultured with different concentrations of test drugs, with HBsAg,HBeAg and HBV DNA in the cultured supernatant determined by time-resolved immunofluorometric assay and fluorescent quantitation PCR assay on day 6th and 10th day,meanwhile,with the cytotoxicity of DNJ derivatives determined by MTT assay.RESULTS:No cytotoxic effects was noted when the test concentration of P-DNJ and NN-DNJ was within 5～125?g?mL-1,HBsAg and HBeAg levels and HBV DNA secretion decreased significantly at a concentration of 125?g?mL-1.CONCLUSION:P-DNJ and NN-DNJ showed anti-HBV activity in the in vitro cell culture experiment.

Objective:To observe the sensitivity of transcription mediated amplification (TMA),and to compare its performance with real-time reverse transcription polymerase chain reaction (real-time RT-PCR) in detecting human immunodeficiency virus RNA (HIV RNA).Methods:TMA system was established with TaqMan probes,specific primers,moloney murine leukemia virus (MMLV) reverse transcriptase,T7 RNA polymerase,and reaction substrates.The sensitivity of TMA was evaluated by amplifying a group of 10-fold diluted HIV RNA standards which were transcribed in vitro.A total of 60 plasma of HIV infected patients were measured by TMA and Cobas Amplicor HIV-1 Monitor test to observe the positive rate.The correlation and concordance of the above two technologies were investigated by linear regression and BlandAltman analysis.Results:TMA system was established successfully and HIV RNA transcribed standards at concentration of equal or more than 10 copies/mL could be detected by TMA technology.Among 60 samples of plasma from HIV infected patients,46 were positively detected and 12 were negatively amplified by both TMA and Cobas reagents;2 samples were positively tested by Cobas reagent but negatively tested by TMA system.The concordance rate of the two methods was 97.1％ and the difference of positive detection rate between the two methods was not statistically significant (P＞0.05).Linear regression was used for 46 samples which were positively detected by both TMA and Cobas reagents and showed an excellent correlation between the two reagents (r=0.997,P＜0.001).Bland-Altma analysis revealed that the mean different value ofHIV RNA levels for denary logarithm was 0.02.Forty-four samples were included in 95％ of credibility interval of concordance.Conclusion:TMA system has the potential of high sensitivity.TMA and real-time RT-PCR keep an excellent correlation and consistency in detecting HIV RNA.

Objective To investigate clinical features and antifungal therapeutic effect of chronic severe hepatitis (CSH) patients with invasive fungal infection (IFI), and to improve the diagnosis and treatment.Methods Clinical manifestation, blood routine, imageology and mycetology characteristic, antifungal treatment perscription and therapeutic effect of 79 CSH patients with IFI were retrospectively analyzed. Antifungal therapeutic effect was compared between fluconazole and voriconazole. Results Thirteen (16.5%) patients received glucocorticoid or other immunodepressants for a relatively long time, 40 (50.6%) patients had invasive operation, and 61 (77.2 %) patients were administered 1-6 kinds of broad-spectrum antibiotics. Seventy-three patients had fever. Leucocytes and neutrophilic granulocyte increased in 96.2% of the patients. Lung (31.6%), intestinal tract (26.2%) and oral cavity (14%) infections were common. Fungus was found in 70.9% of the patients. Candida albicans (40.9%) and aspergillus (21.1%) were often seen. Halo signs and crescent signs on lung CT were relatively specific in 40% of the patients with fungal pneumonia. Voriconazole was more effective than fluconazole(71.4% vs. 39.0%, P<0.05). Twelve patients with lung aspergillus infection were administered voriconazole, 8 (66.7%) patients of whom was effective, and the other 4 (33.3%) patients died. Conclusion There are high risk factors in major CSH patients with IFI. The most common clinical manifestations of CSH patients with IFI are fever, leukocytosis, lung and intestinal tract infection. Candida albicans and aspergillus infection are common. Voriconazole is more effective than fluconazole, and can increase the survival rate of CSH patients with IFI.

Numerical simulations of volume conduction can be used to analyze the process of energy transfer and explore the effects of some physical factors on energy transfer efficiency. We analyzed the 3D quasi-static electric field by the finite element method, and developed A 3D coupled field-circuit model of volume conduction basing on the coupling between the circuit and the electric field. The model includes a circuit simulation of the volume conduction to provide direct theoretical guidance for energy transfer optimization design. A field-circuit coupling model with circular cylinder electrodes was established on the platform of the software FEM3.5. Based on this, the effects of electrode cross section area, electrode distance and circuit parameters on the performance of volume conduction system were obtained, which provided a basis for optimized design of energy transfer efficiency.
Algorithms ; Electric Conductivity ; Electrodes ; Electromagnetic Fields ; Equipment Design ; Finite Element Analysis ; Models, Theoretical ; Prostheses and Implants ; Skin Physiological Phenomena

Objective:To investigate the current situation of cardiopulmonary bypass(CPB) in China and analyze the causes, to guide the formulation and implementation of technology standard.Methods:The survey task force sent out a nationwide survey to obtain up-to-date information on perfusion practice by ChSECC(Chinese Society of Extracorporeal Circulation). The unit of analysis for the survey was the medical center performs CPB. The survey consisted 48 questions covering four topics of qualifications, including certification and education, policies and practices, device and equipment, techniques used.Results:There were 540 of the 714 centers for an overall response rate of 76%. According to the annual number of CPB, they were divided into 4 groups: group A(≤50 cases/year), group B(50-100 cases/year), group C(100-500 cases/year) and group D(≥500 cases/year). The response rate of center with more than group D last year was 100%. Most of the perfusionists had certification issued by ChSECC. Although there were more than 80% of group D performed regular training and assessment of perfusionist, the result was still not ideal enough. Low utilization of safety equipment was not depend on the annual operation volume in most of responding centers. Ultrafiltration and blood protection technology had high application rate in group D compared with group A and B.Conclusion:The certification rate of perfusionists are high. Lower the number of annual CPB cases, lower the proportion of regular evaluation and training, and lower rate of standards performance. No matter the amount of CPB, the application rate of safety equipment is not ideal. Higher the number of CPB cases, higher the utilization rate of CPB related technologies.

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.