The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21-CodonPlus (DE3)-RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM<'4B> by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD<,tested serum>/OD<,negative serum>≥2.1- The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.

Objective To review the important points in the preoperative assessment and the surgical technique in precise hepatic pedicle dissection in anatomical hepatic segmentectomy.Methods 104 patients who underwent anatomical hepatic segmentectomy were divided into two groups according to the different surgical approaches adopted in a prospective and non-randomized manner:the precise hepatic pedicle dissection group (the precise group,n=44) and the conventional hepatectomy group (the conventional group,n=60).The perioperative and follow-up data were analyzed.Patients who had primary liver cancer,including hepatocellular carcinoma and intrahepatic cholangiocellular carcinoma,were analyzed separately.Results (1) There was no perioperative death in the two groups.There was no significant differences in blood loss and transfusion between the 2 groups of patients (P=0.069,0.208; t=1.844,1.266).There was a significantly higher rate of vascular inflow occlusion (P=0.001).There were significantly longer periods of vascular inflow occlusion and operative time (P=0.001,0.001; t=3.849,3.574) in the precise group.There was no significant difference in postoperative complications (P=0.988) and the duration of postoperative hospital stay (P=0.509;t=0.662) between the two groups.(2) In patients with primary liver cancer,there were no significant differences between the precise group (n=29) and the conventional group (n=41) in tumor margin positivity,vascular invasion and pathological staging (P=0.985,0.630,0.769).(3) All patients were followed up for two years.When compared with the conventional group,the disease-free survival (P=0.012),overall survival (P =0.006),and median survival (16.5 ± 4.5mo vs.7.8 ± 3.8mo)were significantly longer in the precise group.Conclusion Precise hepatic pedicle dissection had the same safety and efficacy as conventional method in partial hepatectomy.For primary liver cancer,precise hepatic pedicle dissection had better survival compared to the conventional method when the surgical margin was negative.

Objective We analyzed the curative effect of ERCC1 and RRM1 expression on the Neo-adjuvant Chemotherapy of stage Ⅲ NSCLC to investigate the guiding function of ERCC1 and RRM1 expression in chemotherapy regimen containing platinum.Methods Branch DNA-liquid phase chip methods were used to detect ERCC1 and RRM1 expressions before chemotherapy in 80 cases of stage Ⅲ NSCLC confirmed by pathology.All patients received 2 periods Neo-adjuvant Chemotherapy with GP regimen.According to WHO efficacy appraisal standard,the Enhanced Scan of CT showing reaching complete remission or partial remission was effective or stable,otherwise the progression was considered ineffective.Results For the 80 cases of stage Ⅲ NSCLC,the treatment for 20 of the 25 patients with low expressions of both ERCC1 and RRM1 were effective with an effective rate of 80.0％;The treatment for 14 of the 23 patients with low expressed ERCC1 and high expressed RRM1 were effective with an effective rate of 60.9％;The treatment for 10 of the 20 patients with high expressed ERCC1 and low expressed RRM1 were effective with an effective rate of 50.0％;and the treatment for 4 of the 12 patients with both high expression were effective with an effective rate of 33.3％.The difference of effective rates among the four groups had statistical significance ( x2=7.81,P＜0.05 ) with group A having significantly higher rate than the other three groups and group B and group C having significantly higher rate than group D ( P＜0.05 ).Conclusion ERCC1 detection has guiding significance on the regimen selection of NSCLC Neo-adjuvant Chemotherapy.It was worthwhile to use ERCC1 detection widely in the individualized treatment of the stage Ⅲ NSCLC before surgery.

Objective To observe the intluence ot chemokine RANTES influence on cardiac allograft acute rejection caused by alloreactive memory CD4+ T cells (Tm) adoptive transfer.Methods Heterotopic heart transplantation (HTx) from Balb/c donors to C57BL/6 recipients was performed by anastomosis of the vessels of the neck.Mice undergoing heterotopic heart transplantation received either adoptive transfer of 1 × 106 CD4+ Tm from the spleen of alloantigen-primed C57BL/6 mice or no cells (control group).After the cardiac transplantation,the mean survival time (MST),mean histologic rank of rejection,relative gene expression and serum concentration of RANTES in the cardiac grafts.Results (1) The percentage of CD4+ Tm was 26.83％ at the spleen of alloantigenprimed mice; (2) The MST was 5.17 ± 0.17 days in the CD4+ Tm+ HTx group versus 7.76 ± 0.21 days at the HTx group (control group) (P＜0.01); (3) The histological tests revealed that mean histologic rank of rejection activity in the sections of cardiac allografts on the day 5 post grafting was grade 3.92 ± 0.08 in the HTx+ CD4+Tm group versus grade 2.67 ± 0.14 in HTx group (P＜0.01) ;(4) The relative gene expression level of RANTES was 2.6 ± 0.21 in the CD4+ Tm + HTx group,significantly higher than in the control group (P＜0.01) ; (5) The serum concentration of RANTES in the CD4+ Tm+ HTx group was 223.6 ± 16.79 pg/mL,higher than in the control group (120.7 ±9.47 pg/mL,P＜0.01).Conclusion Alloreactive CD4+ Tm contribute to the increased expression and secretion of RANTES,and cardiac allograft acute rejection was more extensive in the CD4 + Tm + HTx group.

Objective To investigate the effects of cochlear implantation on residual hearing and to evaluate the potential impact of long -term electrical stimulations on residual hearing .Methods 58 hearing impaired children with cochlear implants were included in this study .All subjects could cooperate with behavioral audiometry .Audio-metric evaluations were carried out pre -implantation and 3 ,12 ,24 months post -implantation respectively .Of 58 subjects ,43 were followed up more than 1 year and 17 were followed up more than 2 years .Results All 58 subjects showed significant differences (P<0 .05) between pre- and 3 months post-implantation of residual hearing at the individual frequencies of 0 .25 ,0 .5 ,1 ,2 and 4 kHz .43 subjects followed up more than 1 year showed statistic differences (P<0 .05) between pre- and 3 months post -implantation ,pre- and 12 months post-implantation at 0 .25 ,0 .5 ,1 ,2 and 4 kHz respectively .Comparing 3 months with 12 months post -implantation ,there were sta-tistic differences at 0 .25 and 0 .5 kHz ,while no significant difference (P>0 .05) at 1 ,2 and 4 kHz .Of 17 subjects followed up more than 2 years ,there were significant differences (P<0 .05) between pre- and various return visits post-implantation .Post-implantation return visits ,there were significant differences between 3 months and 12 , 24 months at 0 .25 and 0 .5 kHz respectively ,not any significant differences on 1 ,2 and 4 kHz .There were no sig-nificant differences on each frequency between 12 months and 24 months post- implantation .Conclusion Residual hearing after cochlear implantation could decrease to some extent for various reasons .There were significant differ-ences between 3 and 12 months post-implantation at 0 .25 and 0 .5 kHz .Not any significant differences were ob-served between 12 months and 24 months post-implantation at each frequency .

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Cochlear Nerve ; chemistry ; pathology ; Cranial Nerve Neoplasms ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Heart Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Neurilemmoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vestibulocochlear Nerve Diseases ; metabolism ; pathology ; Vimentin ; metabolism

In this study,a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV)E2 glycoprotein was expressed in E.coli to a obtain target protein.This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs.The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve(ROC)analysis based on 30 negative sera and 80 positive samples.The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination(IHA)test.The inter-assay and intra-assay coefficients of variation (CVs)for 16 sera were both ≤6.8%.No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus(BVDV)antibodies was observed.

Objective To develop and evaluate an up-converting phosphor technology-based lateral flow assay ( UPT-LF) for detection of aflatoxin M1(AFM1) in milk powder and milk.Methods AFM1-UPT-LF was established with up-converting phosphor ( UCP) nano-particles as the bio-label of competitive mode based LF assay .Sensitivity, quantitative ability and precision were evaluated using simulated AFM 1-postive samples with serial standard concentrations .The qualita-tive and quantitative detection performance of AFM 1-UPT-LF was evaluated with reference to liquid chromatography-mass spectrography ( LC-MS) to detect samples of milk powder and milk simultaneously .Results AFM1-UPT-LF could conduct qualitative and quantitative detection without sample pretreatment within 20 min.The detection limit of AFM1-UPT-LF reached 0.1 μg/kg in milk powder and 0.3 μg/L in milk.There was good linearity ranging from 0.1 to 0.7 μg/kg and 0.3 to 0.7 μg/L for milk powder and milk, respectively.The sensitivity, specificity and receiver operating characteristic ( ROC) area under the curve ( AUC) of AFM1-UPT-LF for qualitative result could meet the need of national standards for AFM1 limit in dairy products.After statistical analysis, there was no significant difference (milk powder: t=0.66, P>0.05;milk:t=1.01, P>0.05) between AFM1-UPT-LF and LC-MS for quantitative detection .Conclusion The good qualitative and quantitative detection performance of AFM 1-UPT-LF for milk powder and milk makes possible on-site rapid detection of AFM1 in dairy products quantitatively .

Objective To construct a prokaryotic expression system for tlyA gene of Leptospira in-terrogans ( L.interrogans) strain and to investigate the effects of the expressed rTlyA protein on the hemolysis of sheep erythrocytes and the secretion of pro-inflammatory cytokines by human THP-1 cells and murine J774A.1 macrophages.Methods The fragment of tlyA gene of L.inetrrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai was amplified by PCR.The PCR product was sequenced after T-A cloning.A prokary-otic expression system for the tlyA gene was constructed by using pET-42a as the expression vector and E.coli BL21DE3 strain as the host strain.The expression of rTlyA protein was detected by SDS-PAGE.Ni-NTA af-finity chromatography was performed for purification.The lytic activity of the rTlyA protein on sheep erythro-cytes was determined by plate hemolytic test and spectrophotometry.Real-time fluorescence quantitative PCR and Western blot assay were performed to respectively detect the expression of tlyA gene in THP-1 and J774A.1 cells at mRNA and protein levels after infection with L.interrogans strain Lai.ELISA was per-formed to evaluate the effects of the rTlyA protein on the secretion of several pro-inflammatory cytokines ( IL-β, IL-6 and TNF-α) by THP-1 and J774A.1 cells.Results The nucleotide and amino acid sequences of the cloned tlyA gene were 99.2% and 99.8% identical to those corresponding sequences in GenBank, re-spectively.The constructed prokaryotic expression system for tlyA gene successfully expressed the rTlyA pro-tein.The rTlyA protein at the concentration of 10μg/ml showed stronger lytic activity on sheep erythrocytes. The transcription levels of tlyA gene (P<0.05) and the secretion of TlyA protein in THP-1 and J774A.1 cells were significantly increased after infecting with L.interrogans strain Lai for 1 to 8 hours.The rTlyA pro-tein at concentrations of 0.1, 1 and 10 μg/ml significantly enhanced the secretion of IL-1β, IL-6 and TNF-αby THP-1 and J774A.1 cells (P<0.05).Conclusion The TlyA protein, encoded by the tlyA gene of L.interrogans strain, was confirmed to be a hemolysin with the ability to induce macrophages to secret IL-1β, IL-6 and TNF-α.This study suggested that the TlyA protein played an important role in inflammatory responses during leptospirosis.