In this study,we constructed 2 eukaryotic expressing plasmids namely pcS2?S and pFP,encoding HBV preS2+S envelope protein and human IL 2/IFN ? fusion protein, respectively. The double plasmid fusion protein was used as an adjuvant in preparation of a treatment type HBV gene vaccine. To investigate its feasibility for use as a therapeutic DNA vaccine, we've evaluated the immune response after injection into healthy mice, HBV transgenic(Tg) mice, New Zealand rabbits and Rhesus monkeys. The results showed that the therapeutic DNA vaccine can improve: (1)the CTL activity; (2)the HBsAg(30?g/ml) specific T lymphocyte proliferation in which the stimulation index(SI) and cytokines(IL 2/IFN r) release levels of the pS2.S immunized group(SI=5.6?0.9; 226.3?41.1/51.1?7.1pg/ml) were significantly higher( P

The peptides from HBV cytotoxic T lymphocyte(CTL) epitope were used to either stimulate or impact the immune effector (E) or CTL target (T) cells respectively, in order to investigate the cellular immune response induced by DNA vaccination in both healthy and HBV transgenic (Tg) mice. It was found that HBV DNA based immunization could induce CTL activity in healthy BALB/c mice, with intensity correlated with the E/T ratio and IFN ? secretion level in its supernatants. The target cells hit by HBsAg CTL epitope peptide(pp20) could be lysed by the active CTL induced by HBV DNA vaccine encoding preS2 and HBsAg, while the cell lysis could not be observed in the target cells impacted by the HBcAg CTL epitope peptide (pp10). The supernatant IL 12 secretion level (211 3?39 8pg?ml -1 ) in the DNA vaccination group was significantly( P

Objective:To investigate the feasibility and its action of the therapeutic DNA vaccine for treatment of HBV infection.Methods:By genetic recombinant technique,have constructed 2 eukaryotic expressing plasmids namely pS2.S and pFP,encoding HBV envelope middle protein(preS2+HBsAg) and human leukocyte cytokines fusion protein(IL 2/IFN ?) genes respectively and evaluated its efficacy for inducing cellular immunity in healthy BALB/c mice and HBsAg serum conversion in HBV transgenic(Tg) mice after immunization of the plasmids by intramuscular administration.Results:1.The T cell proliferation by in vitro HBsAg stimulation closely correlated with HBsAg concentration,with its stimulation index(SI=5.6?0.9) in pS2.S immunized group,being significantly higher than that (2.0?0.5) of the control.The IL 2 and IFN ? secretion level in supernatant of the DNA vaccine immunized spleen cell culture were significantly higher than that of the control,while the IL 4 secretion level was not affected.2.The dendritic cells(DCs) extracted from the local draining lymph node(LN) after DNA vaccination induced a HBsAg specific T cell proliferation with its SI(4.20) being higher than that(2.55) of the control.3.In each group of high dose pS2.S and the middle dose combined with pFP inoculation,the HBsAg serum conversion occurred in one HBV Tg mouse,with the serum anti HBs level increasing as the time prolonged,while the serum HBsAg level of the rest mice were significantly lower than that of the control.Conclusion:The results suggest that HBV DNA vaccine can effectively induced cellular immunity in healthy mice;and the preliminary results of the immunized HBV Tg mice may provide an experimental evidence for further investigation of DNA vaccine for its use in treatment of HBV infection.

Glutathione S-transferase (GST) gene, PcgstA was cloned from the penicillin producing strain Penicillium chrysogenum,which is important for understanding the industrial fermentation process. PcgstA gene has an open-reading-frame of 840 bp in length,which is interrupted by two introns. The deduced amino acid sequence shows about 50% identity to several characterized filamentous fungi GSTs. The recombinant PcGSTA in Escherichia coli were overexpressed and purified. Enzymatic assays showed that the recombinant PcGSTA had a specific activity with 1-chloro-2, 4-dinitrobenzene of (0.159±0.031) μmol/(min· mg). It was found that the expression level of PcgstA in the penicillin producing medium supplemented with phenylacetic acid, the side chain precursor of penicillin G, was significant down regulated than that in medium without phenylacetic acid. This result suggested that PcGST may be related to phenylacetic acid metabolism in the penicillin producing strain.

Objective To explore the significance of peritubular capillary C4d deposition in histopathological changes, renal function and prognosis of the patients with antibody-mediated chronic rejection (AMCR). Methods Deposition of C4d in the kidney was examined by irnmunohistochemistry on routine paraffin-embedded sections using anti-C4d polyclonal antibody. Seventy-seven patients were divided into C4d+ group (n = 35) and C4d- group (n = 42). The relationship of C4d and renal function,histopathological changes and prognoses of allografts were analyzed. Results The number of patients with tubular atrophy and glomerular basement membrane proliferation in C4d+ group was significantly more than that in C4d group (P＜0.05). Mean serum creatinine level was significantly higher in C4d+ group than in C4d- group 12 months after renal transplantation [(379.1 + 260.2)μmol/L vs (260.5 + 175.3) μmol/L, P＜0.05]. According to Kaplan-Meier analysis, the one-year graft survival rate was lower in the C4d+ group (62.9% ) than in the C4d- group (83.3% ) (logrank P＜0.05). Conclusion Patients with C4d deposition are associated with tubular atrophy and glomerular basement membrane proliferation. The serum creatinine level in C4d+ patients was significantly higher than in C4d- group at the 12th month after transplantation. More patients with C4d deposition lost their grafts during the study period.

Objective To verify the feasibility of discriminating tissue types in the anterior approach of the cervical spine by the Electrical impedance (EI) and decrease the rate of severe complications.Methods Six New Zealand white rabbits and 6 mini-pigs were performed standard anterior cervical surgery.The esophagus,carotid artery,tracheal cartilage,annular ligaments of trachea,longus colli muscle and anterior longitudinal ligament are classified as the prevertebral group;The cortical bone,cancellous bone,anulus fibrosus and nucleus pulposus are classified as the vertebral group.Once all the tissues were exposed completely,over the frequency range of 200-3 000 kHz (the frequency points were 200 kHz,400 kHz,600 kHz,800 kHz,1 000 kHz,2 000 kHz,3 000 kHz),the in vivo EI of prevertebral group and vertebral group were measured by a probe and a precision inductance-capacitance-resistance (LCR) meter;then the data were analyzed with IBM SPSS Statistics 22 software.At every frequency,Kruskal-Wallis test followed by all pairwise multiple comparisons was applied for all the four groups (the prevertebral group of rabbits,the vertebral group of rabbits,the prevertebral group of mini-pigs,and the vertebral group of mini-pigs),respectively.P values ＜ 0.05 were considered statistically significant.Results At every frequency,the results of Kruskal-Wallis test for all the four groups were significant,and the results of multiple comparisons were as follows.(1) In both miniature experimental animals and large experimental animals,the EI of the vertebral group tissues was significantly different at every investigated frequency (multiple comparisons).(2) In the miniature experimental animals,the EI between longus colli muscle and esophagus,carotid artery and annular ligaments of trachea,and tracheal cartilage and anterior longitudinal ligament had no significant difference at any frequency (multiple comparisons);significant difference could not be detected in the EI between anterior longitudinal ligament and annular ligaments of trachea at the frequencies of 2000-3 000 kHz (multiple comparisonsk) and could be detected at the frequencies of 200-1 000 kHz (multiple comparisons);for comparison of all other paired tissues of prevertebral group there were significant differences at all frequencies (multiple comparisonsk).(3) In the large experimental animals,the EI between longus colli muscle and esophagus,carotid artery and annular ligaments of trachea,and tracheal cartilage and anterior longitudinal ligament had no significant difference at any frequency (multiple comparisons),the same was true for the EI between annular ligaments of trachea and esophagus,carotid artery and anterior longitudinal ligament,and longus colli muscle and annular ligaments of trachea at the frequencies of 800-3 000 kHz,3 000 kHz and 3 000 kHz (multiple comparisons),respectively;for comparison of all other paired tissues of prevertebral group there were significant differences at all frequencies (multiple comparisons).Conclusion At certain frequencies,the EI among tissues was significantly different and could discriminate tissues in the anterior approach of the cervical spine.

Percutaneous coronary intervention(PCI) can effectively restore reperfusion of ischemic myocardium in patients with coronary artery disease.But currently PCI treatment is just available for a small proportion of the patients with coronary artery disease,so there was an insufficiency of PCI in patients with coronary artery disease.From the perspective of ethics,the reasons for the phenomenon and some feasible strategies for the problem are also analyzed in this paper.

ObjectiveTo investigate the efficacy of rapamycin combined with CsA/Tacrolimus (Tac) in chronic allograft nephropathy (CAN).MethodsFifty-three cases of CAN accepted the quadruple immunosuppressive drug program,which contained rapamycin combined with CsA/Tac and MMF and prednisone,and CsA/Tac and MMF were reduced to the original amount of 25％ to 50％.After treatment for 12 months,more relevant indicators,including serum creatinine,glomerular filtration rate,serum cholesterol,triglycerides,urinary protein,GPT and bilirubin and other changes were observed.ResultsIn the patients receiving quadruple regimen of rapamycin during 12 months,the blood Ccr was decreased from (161.51 ± 106.48)μmol/L before treatment to (126.51 ± 56.2)μmol/L after treatment for 6 months (P＜0.05) and to (123.43 ± 54.18)μmol/L after for 12 months (P＜0.01).The GFR was increased from (0.754 ± 0.302) ml/s before treatment to (0.952 ± 0.347)ml/s after treatment for 6 months (P＜0.05) and to (1.007 ± 0.394) ml/s after treatment for 12 months (P＜0.01).Cholesterol and triglycerides in patients had no significant change before and after treatment.The positive rate of proteinuria after treatment showed an increasing trend from 9.4％ before treatment to 26.4％ after treatment for 12 months.ConclusionThe quadruple program of rapamycin combined with CsA/FK506 and MMF can significantly improve Ccr and GFR in patients with CAN,but it can increase the incidence of proteinuria in patients:

Objective To analyze the epidemiological features of public health emergencies from 2004 to 2007 in Daoxian,so as to provide scientific basis for effective early warning,prevention and control of public health emergencies of infectious diseases. Methods The descriptive epidemiological analysis was performed based on public health emergencies in Daoxian from 2004 to 2007. Results The two public health emergency events from 2004 to 2007 in Daoxian and 143 persons involved,none was dead. They were all school outbreak of infections diseases (100%),one of them was outbreak of hepatitis A,and the other was outbreak of other infectious diarrhea. All the public health emergencies were controlled well because of effective measures on time. Conelusion To prevent and control public health emergencies effectively,emphasis should be put on strengthening early warning and health management in schools.

Objective To discover more novel bat viruses and molecularly characterize bat-borne bocavirus diversity in Yunnan.Methods Twenty-six Aselliscus stoliczkanus were sampled in Jinghong , Yunnan, and subjected to viral metagenomic analysis.Specific PCR was used to detect any bocavirus in these samples based on the metagenomic result , while full genome was amplified and compared with other bocaviruses .Results and Conclusion Totally, 3 of the 26 (11.5%) bats were positive for bocavirus, the full genome of which contained 5203 nucleotides and could encode NS1, NP and VP1/VP2 proteins.Phylogenetic analysis showed that this virus shared up to 58.7% and 53.3% amino acid identities with canine bocavirus 1 and canine minute virus .According to ICTV criteria (85%amino acid sequence identity ) on a new species of bocavirus , this virus could be a novel species within genus Bocaparvovirus .This study provides important data to better understand viral diversity in bats and to uncover the relationship between bocavirus and its hosts .