Chinese Center for Disease Control and Prevention published three confirmed novel H 7N9 avain influenza virus in-fection cases on March 31 ,2013 .The disease then spread across China and caused extremely high morbility and mortality , which trig-gered extensive attention .These cases were caused by a novel H 7N9 avian influenza virus which was highly mutable .Once the virus a-dapted to human respiratory epithelium receptors , a pandemic outbreak would occur immediately .This article summerizes the research progress on the H7N9 avain influenza, including epidemiologic characteristics , etiology features, clinical manifestation, detection tech-niques as well as theraputic and prevention strategy .

Objective To identify the potential DNA vaccine candidate which can induce the protective immune response to Toxoplasma gondii by inoculating mice with plasmid DNAs encoding three different forms of P30 antigen (membranous secretory,and intracellular). Methods Three forms of recombinant plasmid: pcDNA3 P30Mb(contain the whole P30 gene sequence,including the gene encoding signal peptide and hydrophobic tail),pcDNA3 P30Se(contain the whole P30 gene sequence, without the gene encoding hydrophobic tail) and pcDNA3 P30In(contain the whole P30 gene sequence,without the gene encoding signal peptide) were constructed by PCR and subcloning technique. The mice were immunized with different forms of recombinant plasmids and IgG antibodies in the mice were detected by ELISA and Western blotting. Results Three forms of expression recombinant plasmid of Toxoplasma gondii P30 gene were successfully constructed. The P30 inserts were identified by restrictive enzyme digestion and sequencing. ELISA and Western blotting analysis demonstrated that specific IgG antibody could be induced in three immunized groups, but there was some difference in appearence time and intensity of IgG.Conclusion Genetically immunization of mice with the recombinant plasmids could elicit specific IgG antibodies. In respect to IgG response, the immune efficiency of the three forms of recombinant plasmids was different at the beginning (2 wk),but 4 wk later approximately same.

The partial segment of Marek′s disease virus (MDV) glycoprotein B (gB) gene was amplified by PCR. The segment was cloned into pET-28a vector to obtain the recombinant pET-gB plasmid. The recombinant plasmid was transformed into E.coli BL21,and expressed in very high level as inclusion body after induced with 1.0mmol/L IPTG. The inclusion body was solubilized in urea (8mol/L) . The purified protein was obtained by use of His?Bind affinity chromatography. Mice were immunized i.p. by the purified protein to make the polyclonal antibody. The titer of the antibody by indirect ELISA was 1?10~ -5 . Moreover, the analysis by western blot proved that antibody was specific to the recombinant protein. These works lay a favorable foundation for the study of the immune response by MDV gB.

AIM:To establish an HPLC method for determining the contents of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd. METHODS:The samples were separated on an Alltima C 18 (250 mm? 4.6 mm,5 ?m) column with the mobile phase of MeOH(A)-0.5% glacial acetic acid solution;gradient elution(0～15 min,30%～60% A;15～30 min,60%～60% A).Flow rate was 1.0 mL/min. The detection wavelength was set at 350 nm.Column temperature was at 30 ℃. RESULTS:The contents of caffeic acid,quercetin and campherenol were 14.218～23.695 ?g/g,9.919～25.564 ?g/g and 6.229～18.160 ?g/g in Hedyotis diffusa Willd from different sources. The linear range of caffeic acid was 0.005 0～0.200 0 ?g(r=0.999 9),the average recovery was 102.35%,RSD was 2.31%(n=6);The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 9),the average recovery was 101.84%,RSD was 1.79%(n=6);The linear range of campherenol was 0.007 8～ 0.310 6 ?g(r=0.999 9),the average recovery was 99.04%,RSD was 2.90%(n=6). CONCLUSION:The method for quantifying of caffeic acid,quercetin and campherenol in Hedyotis diffusa Willd is accurate and reliable.

Objective To explore status quo of Shanghai community health services based on the investigation of rural community doctors in Shanghai suburb,and to propose measures on training rural doctors. Methods Retrospective investigation was carried out in 114 rural community health service centers and 1 563 clinics.Primary investigation included the registration,distribution,age,academic titles and education background. Results The results showed insufficient number of human resources and inadequate investment in human capitals in the primary medical services of Shanghai suburbs,and there was a serious shortage of general practitioners.The other problems included older age,insufficient academic qualifications,and without professional titles. Conclusion We should improve the status and treatment of rural doctors.Job training and well-trained professional students should be encouraged for the primary health services.More and more doctors should be attracted to work in community health service centers.General practitioners' personnal information network and general medical training network should be established,and the general medical education system gets improvement.

Objective To study the possible role of bone morphogenetic protein 7(BMP 7) in the TGF ?1 induced tubular epithelial myofibroblast transdifferentiation (TEMT) of cultured renal tubular epithelial cells. Methods The normal rat kidney tubular epithelial cell line (HK 2) was cultured for three days on plastic plates in the presence or absence of recombinant TGF ?1 and BMP 7. The alterations in the phenotype were assessed by phase contrast microscopy. Transdifferentiation of tubular cells into myofibroblasts was assessed by immunofluorescence, with monoantibodies to alpha smooth muscle actin (? SMA), vimentin and cytokeratin respectively. The expression of ? SMA of HK 2 cells was measured by flowcytometry. The expression of ? SMA mRNA of HK 2 cells was assessed with reverse transcriptase polymerase chain reaction (RT PCR). Results Treatment of HK 2 cells with BMP 7(50 and 100 ng/ml) for 24～48 hours increased cellular proliferation. The culture of HK 2 cells in the presence of TGF ?1 induced a clear fibroblast like morphology, a loss of the epithelial marker cytokeratin and de novo expression of ? SMA and vimentin. Immunofluorescence staining showed the addition of various concentrations of BMP 7 to subconfluent cells for 24 and 48 hours, and the expression of ? SMA and vimentin was decreased. There was an increase in the percentage of cells expressing ? SMA with TGF ?1, which was completed inhibited by an addition of BMP 7(P

OBJECTIVETo study the effects of Jisuikang (Chinese characters) on Nogo-NgR gene expression, and to explore the protective effects and mechanism of Jisuikang (Chinese characters) on spinal cord injury in rats.

METHODSOne hundred eighty female rats were randomly assigned to 6 groups(30 rats per group). Sham group: T10 lamina was resected only and spinal cord was untreated. Model group: spine cord injury (SCI) was created with a modified impinger of Allen's by impacting on the T10 spinal cord. Prednisolone group: Prednisolone (0.06 g/kg) was given by intragastric administration at a time interval of 24 hours after operation. The Jisuikang (Chinese characters) high, moderate and low dose groups: Jisuikang (Chinese characters) was supplied with different dose (50 g/kg, 25 g/kg, 12.5 g/kg) by intragastric administration in rats after operation,for the first time at 30 min after surgery. Animals were killed 3, 7, 14 days after surgery. The expression levels of Nogo-A and NgR were observed by Western Blot and Real-time PCR.

RESULTSThe expression of Nogo-A and NgR was at the basic level at all time points in sham group. Compared with model group, the protein expression levels of Nogo-A and NgR in sham, prednisolone, Jisuikang (Chinese characters) moderate dose groups were statistically significant at all time points (P < 0.05). No difference was found in Jisuikang (Chinese characters) high and low dose groups (P > 0.05). Three days after surgery, the mRNA levels of Nogo-A and NgR in treatment group were significantly lower than that in model group (P < 0.01); 7 days after surgery,Nogo-A and NgR mRNA expression were dramatically upregulated and peaked; 14 days after operation, the expression was decreased, but still significantly higher than that in other treatment groups (P < 0.01). Prednisolone and Jisuikang (Chinese characters) moderate dose groups showed the most significant effects among all groups,but there was no statistically significant difference between two groups (P > 0.05).

CONCLUSIONThe decoction Jisuikang (Chinese characters) can promote the nerve cell regeneration by regulating Nogo-A and NgR gene expression, activating Nogo- NgR signaling pathways after acute spinal cord injury.

Animals ; Female ; GPI-Linked Proteins ; analysis ; genetics ; physiology ; Medicine, Chinese Traditional ; Myelin Proteins ; analysis ; genetics ; physiology ; Nerve Regeneration ; drug effects ; Nogo Proteins ; Nogo Receptor 1 ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; analysis ; genetics ; physiology ; Signal Transduction ; drug effects ; Spinal Cord Injuries ; drug therapy ; metabolism

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

Objective To investigate the analgesic efficacy of ultrasound?guided adductor canal blockade (ACB)after minor arthroscopic knee surgery. Methods Sixty patients undergone minor arthroscopic knee surgery were randomly divided into group ACB(n=20)and group Control(n=20). All patients received spinal anesthesia. The patients in group ACB received ultrasound?gGuided ACB with 20 ml 0.5% ropivacaine,and patients in group Control received 20 ml saline after the surgery. In addition ,all patients have a basic analgesic regimen with etoricoxib. Visual analogue scales(VAS) during rest and passive movement ,additional analgesic dose and side effects were recorded at 4,8,12,24 h Post?operation. At 24 h post?operation,the numbers of patients who can walk for 5 meters were recorded. Results VAS during rest and movement at 4 h,8 h and 12 h post?operation in group ACB were significantly lower than those in group Control. And all patients could walk 5m at 24 h post?operation. No headache,nausea and vomiting,urinary retention and other adverse reactions were observed in group ACB. There were four patients who received additional analgesic and one patient vomitted. Conclusions Significant analgesic effect of the ACB could be detected after minor arthroscopic knee surgery ,with less reduction in requirements for supplemental opioids.

The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.