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

Ten compounds were isolated from the artificially induced dragon's blood of Dracaena cambodiana by various column chromatographies on silica and sephadex LH-20 gel. Based on spectral analysis of NMR and MS, their structures were identified as 3, 4-dihydroxyallylbenzene (1), 3', 4', 5'-trimethoxycinnamylalcohol (2), pinoresinol (3), (2R)-7, 4'-dihydroxy-8-methylflavane (4), (2R)-7,4'-dihydroxy-5-methoxy-8-methylflavane(5),(2S)-7,3'-dihydroxy-4'-methoxy-8-methylflavane(6) ,(2S)-4',7-dihydroxy-6, 8-dimethylflavane(7), 4,2',4'-trihydroxychalcone(8), 4,4'-dihydroxy-2-methoxydihydrochalcon(9) and Cambodianin E (10). Antibacterial activity assay showed that compounds 1, 4 and 10 have inhibitory effect on Fusarium oxysporum f. sp. cuben, Fusarium oxysporum f. sp. niveum, Fusarium oxysporum f. sp. vasinfectum and Ralstonia solanacearum.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Dracaena ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fusarium ; drug effects ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Spectrometry, Mass, Electrospray Ionization

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.

Objective To examine the effects of the MEK inhibitor on human pancreatic cancer cells, and to explore the molecular mechanisms. Methods Human pancreatic cancer cell lines CFPANC1, PANC1 and MiaPaCa2 were treated with MEK inhibitor PD98059 or DMSO, the sensitivity was analyzed by an MTT assay, and cell cycle distribution was evaluated by flow cytometry( FCM), The transcriptional level and protein expression of tumor suppressor genes were detected by real-time RT-PCR and western blot respectively. DNA methyltransferase (Dnmt)1, 3a and 3b were also assayed by western blot, The methylation status of the promoter of the p16INK4A gene was assayed by methylation-specific PCR (MSP). Results PD98059 inhibited to various degrees the growth of three pancreatic cancer cell lines, accompanied by G0-G1 cell cycle arrest. PD98059 up-regulated the expression of p16INK4a, p21WAF1, p27KIP1 mRNA, demethylated the hypermethylation status of p16INK4a gene promoter, and decreased Dnmtl and Dnmt3b in CFPANC1 and PANC1 cell lines. PD98059 only increased the expression of p27KIP1, while the changes of p16INK4a, p21WAF1 and Dnmt were less marked in MiaPaCa2 cell line. Conclusions MEK inhibitor PD98059 down-regulate the activation of Dnmt and up-regulate tumor supress genes, along with the inhibition of cell proliferation and cell cycle progression.

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.

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

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 .

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.