Objective To study the effect of prolactin(PRL) on the activation of T lymphocytes stmiulated by concanavalin A(ConA),and to explore the action of PRL in the activation of T lymphocytes. Methods After CD4 +T cell line JurkatE6-1 cells were respectively stmiulated by 5 mg/L ConA,25 ?g/L PRL and 500 ?g/L bromocriptine(Brc).The blank control group,the ConA group,the PRL and ConA group(PRL group),the Brc and ConA group(Brc group),the PRL and Brc group(PRL-Brc group) were set in the experiment.The total RNA was extracted by Trizol after 48 hours and was reversed transcription immediately.The expression of tumor necrosis factor receptor associated factor 6(TRAF6) mRNA of T lymphocytes was checked by PCR.The expressions of tumor necrosis factor(ligand) super family 4(TNFSF4) and Killer specific secretory protein of 37 000(KSP37) mRNA of T lymphocytes were detected by real-time polymerase chain reaction. Results The PRL group and the Brc group could inhibit the expressions of TRAF6,TNFSF4,and KSP37 mRNA of the activated T lymphocyte compared with the blank control group and the ConA group(P a0.05).The PRL-Brc group could inhibit significantly the expressions of TRAF6,TNFSF4,and KSP37 mRNA of the activated T lymphocyte compared with the ConA group(P a

The discovery of microRNAs (miRNAs) has introduced a new paradigm into gene regulatory systems. Since inception, computational methods have been an invaluable tool complementing experimental approaches, and many discoveries have been obtained through combination of experimental and computational approaches. The knowledge that has been accumulated about the principles of miRNAs and target recognition were reviewed. The currently available computational methodologies and software for prediction of miRNA and their target genes also have been discussed.

OBJECTIVETo evaluate the application of side-to-side anastomosis of the lesser curvature of stomach and jejunum in laparoscopic Roux-en-Y gastric bypass (LRYGB).

METHODSClinical data of 29 patients with type 2 diabetes mellitus (T2DM) undergoing side to side anastomosis of the lesser curvature of stomach and jejunum in LRYGB from May 2012 to November 2012 in Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University were analyzed retrospectively.

RESULTSAll the procedures were successfully completed without conversion to laparotomy. The side-to-side anastomosis of the lesser curvature of stomach and jejunum avoided the laparoscopic suture. No gastrojejunostomy anastomotic bleeding, fistula, obstruction and other complications occurred after operation and no complications of gastrojejunostomy anastomosis were found during a follow up of 1 to 7 months.

CONCLUSIONSSide-to-side anastomosis of the lesser curvature of stomach and jejunum in LRYGB can manipulate the size of anastomosis accurately and avoid the laparoscopic suturing. It is simple and easy to learn.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Follow-Up Studies ; Gastric Bypass ; methods ; Humans ; Jejunum ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach ; surgery ; Treatment Outcome

OBJECTIVETo construct a human phage display antibody library, which will help to develop new drugs and vaccines against respiratory syncytial virus (RSV) and solve many of the issues that have limited the progression and application of murine monoclonal antibodies (McAbs) in the clinic. This can provide a platform for human antibody preparation and diagnosis, prophylaxis and therapy of RSV infection in children.

METHODSPeripheral blood lymphocytes were isolated from 52 children with RSV infection. cDNA was synthesized from the total RNA of lymphocytes. The light and heavy chain Fd (VH-CH1) fragments of immunoglobulin gene were amplified by RT-PCR. The amplified products were cloned into phagemid vector pComb3x and the clone samples were electrotransformed into competent E.coli XL1-Blue. The transformed cells were then infected with M13K07 helper phage to yield recombinant phage antibody of Fabs. The plasmids extracted from amplified E.coli were digested with restriction endonucleases Sac I, Xba I, Spe I and Xho I to monitor the insertion of the light or heavy chain Fd genes. RSV virions were utilized as antigens to screen Fab antibodies.

RESULTSBy recombination of light and heavy chain genes, an immune Fab phage display antibody library against RSV containing 2.08x10(7) different clones was constructed, in which 70% clones had light chains and heavy chain Fd genes. The capacity of Fab phage antibody gene library was 1.46x10(7) and the titre of the original Fab antibody library was about 1.06x10(12) pfu/mL. The antibody library gained an enrichment in different degrees after the preliminary panning.

CONCLUSIONSUtilizing the technology of phage display, an immune Fab phage display antibody library against RSV was successfully constructed in this study, which laid a valuable experimental foundation for further study and created favorable conditions for preparing human McAbs. This may also contribute to the improvement in the diagnosis, therapy and prophylaxis of RSV infection in children.

Antibodies, Viral ; genetics ; Bacteriophages ; genetics ; Child ; Humans ; Immunoglobulin Fab Fragments ; genetics ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Peptide Library ; Respiratory Syncytial Viruses ; immunology

OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.

OBJECTIVETo study the expression of miRNA-106a gene in esophageal squamous cell carcinoma (ESCC) and its association with clinicopathological features and prognosis of ESCC patients.

METHODSReal-time fluorescence quantitative polymerase chain reaction (PCR) assay was used to determine the expression of miRNA-106a gene in esophageal cancer tissue and corresponding normal mucosa of 81 cases. Immunohistochemical technique was applied to detect the expression of p53, human epidermal growth factor receptor 2 (HER-2), DNA topoisomerase II (Topo II) and multidrug resistance-associated protein (MRP). The association of miRNA-106a expression with clinicopathological features, expression of related proteins, and prognosis of the patients was analyzed.

RESULTSAmong the 81 cases, under-expression of miRNA-106a gene was found in 48 cases (59.3%), normal expression in 22 cases (27.2%), and overexpression in 11 cases (13.6%). The expression of miRNA-106 gene was significantly associated with lymph node metastasis, pathological stage, and nerve invasion (all P < 0.05), significantly associated with expression of p53 (P = 0.006), and not significantly associated with expressions of HER-2, Topo II and MRP proteins (all P > 0.05). The expression of miRNA-106a gene was also significantly associated with progression-free survival (PFS, P = 0.032), but not significantly with overall survival (OS, P = 0.486). The results of Cox multivariate regression analysis showed that the PFS of ESCC patients was significantly correlated with lymph node metastasis (P = 0.029), but not correlated with the age, gender, tumor length, T stage, degree of differentiation, nerve invasion, and miRNA-106a expression (all P > 0.05).

CONCLUSIONSIn esophageal squamous cell carcinomas, the miRNA-106a gene is under-expressed, with tumor suppressor function, and may be regarded as a biological marker to assess the prognosis in patients with esophageal squamous cell carcinoma.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; DNA Topoisomerases, Type II ; metabolism ; Disease-Free Survival ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Multidrug Resistance-Associated Proteins ; metabolism ; Neoplasm Invasiveness ; Neoplasm Staging ; Proportional Hazards Models ; Real-Time Polymerase Chain Reaction ; Receptor, ErbB-2 ; metabolism ; Survival Rate ; Tumor Suppressor Protein p53 ; metabolism

Cardiac Catheterization ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Prosthesis Implantation ; instrumentation ; Septal Occluder Device

OBJECTIVETo investigate the changes of blood corpuscles of patients with severe acute respiratory syndrome (SARS) in Beijing.

METHODSTotally 43 patients (21 male and 22 female, 19-74 years old age range) diagnosed as of probable SARS were included in this study. Their corpuscles in the peripheral blood were tested every two days, and the results were analyzed.

RESULTSPatients with SARS were more likely to develop leukocytopenia, lymphopenia and thrombocytopenia in the early period of disease than those in control group. The situation, especially lymphopenia and thrombocytopenia, could not be reversed in patients who died. Persistent low counts of lymphocytes and platelets at presentation might be associated with adverse outcomes.

CONCLUSIONLow counts of leukocytes, lymphocytes and platelets were common among patients in the early stage of SARS. Persistent Lymphopenia and thrombocytopenia may be associated with the prognosis.

Adult ; Aged ; Early Diagnosis ; Female ; Humans ; Leukocyte Count ; Leukocytes ; cytology ; Male ; Middle Aged ; Prognosis ; Severe Acute Respiratory Syndrome ; diagnosis

OBJECTIVETo analysis the clinical and laboratory characteristics of Patients infected with new influenza A (HIN1) virus.

METHODSAll cases with new influenza A (H1N1) confirmed on polymerase chain reaction assay on throat swabs. There were included in a prospective evaluation of clinical characteristics, laboratory results, treatment and overcome of new influenza A (H1N1).

RESULTSThere were 35 patients in the epidemic. Clinical illness developed within a mean of 1.7 days. Fever occurred in 97.1%, sore throat 65.7% cough 51.4%, headache 28.6%, and myalgia 31.4%. All patients were treated with oseltamivir lasted 5 days. The mean duration of viral shedding was 4.5 days. All were cured and left hospital after day 7.

CONCLUSIONIt was infected by new influenza A (H1N1) typically in this epidemic.

Adolescent ; Adult ; Disease Outbreaks ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; pathogenicity ; Influenza, Human ; diagnosis ; drug therapy ; virology ; Male ; Oseltamivir ; therapeutic use ; Young Adult