Objective:To construct and evaluate T7 phage display cDNA library from synovium of rheumatoid arthritis patients.Methods:Total RNA was extracted from pooled RA synovium by Trizol reagents.Messenger RNA was isolated from total RNA by oligo (dT)-conjugated Oligotex particles,and then,the agarose gel electrophoresis showed the range of mRNA size.After mRNA was reverse-transcribed into double-stranded cDNA,end modification,adaptors ligation and EcoR Ⅰ and Hind Ⅲ digestion were performed.After eliminating excess adaptors and small fragments(less than 300 bp),the cDNA was ligated into T7Select 10-3 vector.The RA synovium phage display cDNA library was constructed by package reaction in vitro and plate proliferation.Plaque assay and PCR were used to evaluate the library.Results:We showed that complexity of the library was about 2?107,and the phage titer of the amplified library was 8.9?10 10 pfu/ml.Insert ratio was proved to be 90% with the range of 300-2 000 bp inserts.Meanwhile,a target cDNA gene of BiPwas probed with PCR amplification.Conclusion:The phage display cDNA library from synovium was construted with high quality and can be used as a valuable source in screening of RA self-antigens.

Objective This study is to investigate cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway gene expression pattern of peripheral blood mononuclear cell(PBMC) of primary sjgren syndrome patients.Methods The PBMC sample of 3 patients with sjgren syndrome and 3 healthy volunteers with consistent age were collected.The total RNAs was extracted from the PBMC samples,and reverse transcripted in vitro transcription(IVT),labeled with Cy5/Cy3 and hybridized on the gene chips.After scanning and data extraction with LuxScan 3.0,differentially expressed genes were analyzed with SAM method.The online tool of molecule analysis system(MAS) was used for biological knowledge mining.Results Statistical difference was calculated between the patient and control group in the following three pathways: cytokine pathway,Jak-Stat signal pathway,neuroactive ligand-receptor pathway.Among these,genes of IL-2RA,IL-10 were up-regulated and genes of PF4,GZMA were down-regulated.Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjgren′s syndrome.And the research may provide new target therapy for SS.

Tolvaptan is a novel oral selective arginine vasopressin V2 receptor antagonist.Tolvaptan improves heart failure signs and symptoms without serious adverse events.Tolvaptan has no effect on all-cause mortality and cardiovascular death or admission rate for heart failure.But in heart failure patients with hyponatremia,tolvaptan can decrease cardiovascular death and admission rate for heart failure.

Dust ; analysis ; Humans ; Magnoliopsida ; Pneumoconiosis ; epidemiology ; Prevalence

10.3969/j.issn.2095-4344.2013.26.023

Objective To evaluate the clinical significance of antiplatelet antibody in patients with systemic lupus erythematosus complicated with thrombocytopenia.Methods Antiplatelet antibody (anti-GP Ⅱb/Ⅲa antibody, anti-GP Ⅰb/Ⅸ antibody, anti-GP Ⅰa/Ⅱ a antibody, anti-GP Ⅳ antibody) were detected by modified antigen capture ELISA. The positive rate of antiplatelet antibody between SLE complicated with thrombocytopenia group and without thrombocytopenia group before therapy were compared,and the positive rate of antiplatelet antibody before therapy and after therapy in SLE complicated with thrombocytopenia were compared,and the relevance between antiplatelet antibody and conditions in SLE complicated with thrombocytopenia were analyzed. Rank test and Chi square test were used for statistical analysis. Results The positive rate of anti-GP Ⅱb/Ⅲa antibody and anti-GP Ⅰb/Ⅸ antibody in SLE complicated with thrombocytopenia group before therapy was 50% and 67% respectively, however,the positive rate in SLE without thrombocytopenia group before therapy was 11% and 28% respectively,there was significant difference between the two groups (P＜0.05) and the positive rate of anti-GP Ⅱb/Ⅲa antibody and anti-GP Ⅰb/Ⅸ antibody in SLE complicated with thrombocytopenia group after therapy was 6% and 28% respectively, which was significantly lower than those before therapy (P＜0.05). In SLE complicated with thrombocytopenia group before therapy, there was significant relevance between anti-GP Ⅱb/Ⅲ a antibody and anti-GP[b/Ⅸ antibody, and there was significant relevance between these two antibodies and SLEDAI score,but no significant relevance between these two antibodies and ANA,dsDNA, ANCA. Neither anti-GPⅣ antibody nor anti-GP Ⅰ a/Ⅱ a antibody was detected in patients of this study. Conclusion The positive rate of antiplatelet antibody (anti-GP Ⅱb/Ⅲ a antibody, anti-GP Ⅰb/Ⅸ antibody) is significantly higher in patients with active systemic lupus erythematosus complicated with thrombocytopenia,and these two antibodies are significantly associated with clinical outcomes.

ObjectiveTo investigate the gene expression profiling of articular chondrocyte and the function and pathway of the differentially expressed genes in patients with osteoarthritis (OA) by using gene microarray technique.MethodsThree patients with OA,three patients with rheumatoid arthritis(RA) and three traumatic controls without arthritis were selected and were divided into OA versus RA and OA versus the traumatic control groups,and their articular cartilage cells were cultivated.The gene expression profiling was performed by human genome oligonucleotide microarray technique.The differences of gene expression of the articular chondrocyte in OA versus RA group and OA versus the traumatic control group were compared respectively by two class unpaired test using significant analysis of microarray software.The function and pathway of these differentially expressed genes were analyzed by using the database of Molecule Annotation System.ResultsThe number of differentially expressed genes was 145 when OA compared with the traumatic control,in which 70 were up-regulated and 75 were down-regulated; and the number was 281 when OA were compared with RA,in which 94 were up-regnlated and 187 were down-regulated.The Gene Onto-logy (GO) functions of the differentially expressed genes of OA in each group were related to pathological and immune courses including cellular process,physiological process,cell division,biological regulation and cell signal transduction.The statistically significant pathways of these genes in each group included apoptosis pathway,cell cycle pathway,P53 signaling pathway,Fas signaling pathway,NO pathway,apeptotic cascade pathway,focal adhesion pathway, ECM-receptor interaction pathway, tight junction pathway, adhesive bonding pathway,actin cytoskeleton regulation pathway,bone remodeling pathway,chondroitin sulfate biosynthesis pathway,Jak-STAT signaling pathway,Wnt signaling pathway,Toll recepor signaling pathway,cell adhesion molecules pathway,T cell receptor signaling pathway,and s o on (P＜0.05).They displayed not only marked dif-ferences in GO function and gene pathway of differentially expressed genes between OA versus RA group and OA versus the traumatic control group,but also displayed the significant overlapping of the differentially expressed genes and pathways between the two groups.ConclusionThe differentially expressed genes and pathways of articular chondrocytes involve apoptosis,extracellular matrix and cytokines in OA,which contri-bute to further study of early warning genes of OA.

Objective To investigate signal transduction and cytokine gene expression pattern in peripheral blood mononuclear cell (PBMC) between primary Sjogren syndrome patients and healthy controls. Methods The PBMC of 3 patients with Sjogren syndrome and 3 healthy volunteers with consistent age were collected. The total RNA extracted from the PBMC was carried the reverse transcription and in vitro transcription (IVT), then labeled with Cy5/Cy3 and hybridized on the gene chips. After scanning and data extraction with LuxScan 3.0, differentially expressed genes were analyzed with SAM method. The online tools of molecule analysis system (MAS) was used for biological knowledge mining. The difference of signal transduction and cytokine gene expression of each patient and control were compared. Results Statistical difference was calculated between the patient and control group in the following four pathway: the chemokine-cytokine receptor interaction pathway[(FASLG(Fas ligand), CCL5, CCR3, IL-4R, CXCL10 (CXC chemokine ligand 10), TNFRSF19L (tumor necrosis factor supeffamily 19 ligand), CX3CR1 (CX3C chemokine receptor 1)], adhension molecule pathway [interleukin adhesion molecules-1 (ICAM-1)], Jak-STAT signal pathway [STAT4 (signal transducer and activation of transcription 4), CISH (chromogenic in situ hybridization), IL-4R], Toll-like receptor signal pathway(CCL5, CXCL10). Among these, 4 genes were up-regulated and 6 genes were down-regulated. Conclusion Understanding of differently expressed genes should help us disclose the potential molecular mechanism underlying the development process of pathogenesis of primary Sjogren syndrome.

Background and purpose:Lung cancer is the leading cause of cancer-related deaths worldwide. Approximately 15%of all histological types consist of small cell lung cancer (SCLC). Chemotherapy is one of the major treatment methods. Though the current front-line standard chemotherapy regimen for SCLC is active in most SCLC cases, however, the disease recurs shortly after the ifrst successful treatment with multi-drug resistance phenotype. Our previously study showed that zinc ifnger protein X-linked (ZFX) was overexpressed in SCLC cells. This study aimed to investigate the expression of ZFX in SCLC tissues, and to clear their possible associations with clinical parameters and provide basis for therapy of SCLC. Methods:A total of 98 surgical specimens of small cell lung cancer were collected. The expression of ZFX was examined by quantiifcational real-time polymerase chain reaction in 78 specimens taken from patients with complete clinical data. Results:The expression of ZFX was signiifcantly increased in extensive stages than in limited stages. The expression of ZFX was associated with tumor stage, the sensitivity of chemotherapy, and survival times (all P<0.05), no data was found correlated with gender and ages (P>0.05). Conclusion: ZFX expression might be associated with the development of SCLC, and may be a potential prognosis predictor.

This paper collected data of clinical application of the fosfomy-cin.And the retrospective analysis and introduction was carried out.In order to the clinician recognized the characteristic of “old drug new” of fosfomycin.