Objective To evaluate efficiencies of 4 kinds of γ‐interferon release assay(IGRA) detection kits in diagnosis of pul‐monary tuberculosis .Methods 4 kinds of IGRA reagents produced by Oxford Immunotec Ltd (Oxford) in British ,Shanghai Fuxing Changzheng Medical Science Co .,Ltd(Fuxing) ,Cellectis in Australia(Cellectis) and Haikou VTI Biological Institute(VTI) ,respec‐tively ,were used to determine release levels of peripheral bloodγ‐interferon which had antigenicity of Mycobacterium tuberculosis in 86 cases of patients with tuberculosis and 80 cases of healthy individuals ,and diagnostic efficiencies were evaluated .Results Among the 4 kinds of IGRA reagents ,including Oxford ,Fuxing ,Cellectis and VTI ,the sensitivity was 93 .02% ,88 .37% ,90 .70% and 91 .86% ,respectively ;the specificity was 92 .50% ,75 .00% ,82 .50% and 87 .50% ,respectively ;the positive predictive value was 93 .02% ,79 .17% ,84 .78% and 88 .76% ,respectively ;the negative predictive value was 92 .50% ,85 .71% ,89 .19% and 90 .91% ;the diagnostic accordance rate was 92 .77% ,81 .93% ,86 .75% and 89 .76% ,respectively ;the area under receiver operating charac‐teristic(ROC) curve was 0 .975 ,0 .892 ,0 .958 and 0 .963 .Conclusion There are no significant differences among Oxford ,Fuxing , Cellectis and VTI reagents ,and reagents could be selected according to clinical requirements .

Objective To analyze and evaluate neoadjuvant chemotherapy's value and significance in combining with surgical treatment for limited small cell lung cancer(LD-SCLC).Methods A total of 94 LD-SCLC patients underwent complete resections combined with chemotherapy between January 2000 and January 2011 in Shanghai Pulmonary Hospital.Among these cases,initial two cycles of neoadjuvant chemotherapies were performed for all pathologically confirmed patients (Group A),and initial operations followed by adjuvant chemotherapy were administered to patients without pathology (Group B).The survival rate was analyzed by log-rank test and Kaplan-Meier method.Multivariate analysis of the prognostic factors was performed using Cox's regression model.Results Group A included 43 cases and Group B included 51 cases.The mean age was (56.37 ± 10.18) years.According to the 6th edition of Tumor,Node,Metastasis(TNM) classification of lung cancer,54 cases were at stage Ⅰ or Ⅱ,40 cases were at stage Ⅲ.Overall 5-year survival(5-YS) was 27％.The 5-YS for patients with stage Ⅰ-Ⅱ was notably better than that of stage Ⅲ (34％ vs 20％,P =0.033).For patients with stage Ⅲ,the 5-YS of Group A was significantly better than that of Group B(34％ vs 12％,P =0.020),besides median overall survival for Group A and Group B were 46 and 15 months(P =0.009).Furthermore,the results of multivariate analysis showed that neoadjuvant chemotherapy,surgery and histopathology of SCLC were independent factors that strongly affected survival and prognosis.Conclusion In combined surgical treatment for LD-SCLC,neoadjuvant chemotherapy obviously improved the prognosis of patients with stage Ⅲ.Therefore,it was very important and necessary that pre-surgical neoadjuvant chemotherapy was administered to resectable stage Ⅲ LD-SCLC patients.

OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome

ObjectiveTo explore the expression and clinical significance ofaquaporin (AQP) 1 and AQP 4 in human pulmonary adenocarcinoma H1299 cell line.MethodsH1299 cell line in human pulmonary adenocarcinoma(pulmonary adenocarcinoma group) were obtained,the expressions of AQP1 and AQP4 in mRNA level and their locations were determined in H1299 cell line respectively by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The migration of tumor cells were observed by Matrigel invasion assay.Then normal tissues adjacent of pulmonary adenocarcinoma (above cancer line 3 cm,no tumor cell with pathological proven) were as control group.ResultsThe results of RT-PCR showed that AQP1,AQP4 mRNA was 1.030 ± 0.070 and 1.140 ± 0.190 in conlrol group,which were lower than those in pulmonary adenocarcinoma group (2.021 ± 0.250 and 2.180 ±0.180)(P＜0.05 ).The results of Western blot showed AQP1,AQP4 located on the membrane of H1299 cell.Both AQPI and AQP4 mRNA expressed very high in pulmonary adenocarcinoma group,while expressed very low in control group (P＜0.05).Matrigel invasion assay showed that the invasion was positively related to AQP1,AQP4(r =0.351,P ＜ 0.05 ).ConclusionAQP1,AQP4 significantly over express in H1299 cell line,both of them phy important roles in the growth of tumor tissue and cell migration.

Objective To establish a suitable evaluation index system to track implementation effect of clinical research program.Methods Delphi method was used to creat the evaluation index system.The weighted average method was adopted to determine the weight of each index.Results After two rounds of expert consultation,twenty seven evaluation indices were selected,including three first-class indices,eight second-class indices and sixteen third-class indices,and the weight of each index was determined.Conclusions The evaluation index system reflects the purpose of tracking clinical research to a certain extent.This index system is simple and easy to be used.

Objective To evaluate global and segmental right ventricular ( RV ) systolic functions in patients with excessive volume or pressure load using real‐time three‐dimensional echocardiography ( RT‐3DE) . Methods Forty‐five patients with RV volume overload ,45 patients with RV pressure overload and 45 healthy subjects were underwent RT‐3DE . RV global and segmental ( inflow ,body ,outflow ) end‐diastolic volume (EDV) ,end‐systolic volume (ESV) ,stroke volume (SV) and ejection fraction (EF) were analyzed with TomTec software . The correlations between EF with the three‐dimensional method and two‐dimensional parameters including right ventricle systolic pressure( RVSP) were discussed . Results Global EDV and ESV increased significantly in both patient groups compared with controls ( all P < 0 .05) ,but there was no difference between two patient groups ( P >0 .05) .Compensated increase of SV was found in sixty percent of patients with volume overload but none with pressure overload ( P < 0 .05) . Global EF decreased significantly in both of patient groups (all P <0 .05) ,which was more significant patients with pressure overload ( P < 0 .05 ) . Different patterns of the regional dysfunction were found among the different RV segments . No correlation was found between RVSP and global or segmental EF in patients with pressure overload . Conclusions RT‐3DE could be used to assess global and segmental RV systolic function in patients with pressure and volume overload .

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.

Objective To investigate the expression and significance of Smad4 in peripheral hepatocytes of lesions in mice with hepatic alveolar echinococcosis.Methods Eight mice in the test group were inoculated with alveolar echinococcosis and 8 mice in the control group were injected with normal saline.The expression of Smad4 protein in the hepatic tissue of the mice was detected by immunohistochemistry method,and the data were analyzed by chi-square test.The effect of alveolar echinocoeeosis on the expression of Smad4 protein was investigated.Results Smad4 was detected in cell nuclei and partly in the cytoplasm.Six months after the establishment of the mice model for alveolar echinococeosis,the expression of Smad4 in the hepatic tissue in the test group was significantly higher than in the control group(x2=19.869,P＜0.05).The number of Smad4 with positive expression in the hepatocytes in the test group was slightly higher than in the control group,and the expression intensity of Smad4 in the test group was greater than in the control group(x2=58.3 10,P＜0.05).Conclusions The increase of the expression of Smad4 protein in the periphery hepatocytes and tissues of hepatic alveolar echinococcosis lesions may play a role in hepatic cirrhosis and immune evasion.

· AIM:To compare the effect of fixation and select the optimal fixation solution and condition for PAS staining of guinea pig eyes among three different fixation solution:4％ paraformaldehyde solution,4％ glutaraldehyde solution and Davidson solution.· METHODS:Totally 30 healthy guinea pigs were divided into 6 groups:Group Ⅰ-Ⅱ were fixed in 4％ paraformaldehyde solution and 4％ glutaraldehyde solution for 24h,respectively;Group Ⅲ-Ⅴ were fixed in Davidson solution for 3,6 and 24h,respectively;and Group Ⅵ were fixed in Davidson solution for 3h and then transferred into 10％ neutral formaldehyde solution for 48h.All groups were sectioned by routine section method and undergone PAS staining,and then observed by light microscope.· RESULTS:It was found that the group which was fixed by Davidson solution for 3h,remained the most complete structure for PAS staining (Group Ⅲ).While the effect of fixation for the group which was transferred into 10％ neutral formaldehyde solution for preserving for 48h after fixing in Davidson solution for 3h was also acceptable for PAS staining (Group Ⅵ).The retinal cells remained clear and in order for both groups which was mentioned above.· CONCLUSION:The best fixation condition for PAS staining for eyes of guinea pigs is fixation in Davidson solution for 3h among these fixation conditions,while it is also suitable to transfer the eyes into neutral formaldehyde solution after fixing in Davidson solution for 3h for preserving for long periods,which is not severely reduced the effect of fixation for PAS staining.