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.

Objective To investigate the effect of lemon peel extracts(LPE) on the activity of lactate dehydrogenase and sucrase of Streptococcus mutans (Sm). Methods After serial dilution with trypticase soy broth (TSB) medium containing 2% glucose, LPE was used as the experimental group, and TSB without LPE as the control group. Sm was added to each group,which was then cultured for 6,18,24 and 48 hours in the anaerobic tank. The activity of lactate dehydrogenase (LDH) was measured with the method of oxidation of reduction coenzyme I and the pH value of the culture solution was also detected. The activity of the sucrose was determined with the method of coloration of 3,5-dinitrosalicylic acid. Results The activity of LDH, sucraae and the changes of solution pH were decreased with the increase of the concentration of LPE (P ＜ 0. 01 ). The activity of LDH were declined from (0. 8025 ± 0. 0913 ) × 103 U/L to (0. 2099 ±0. 0283) × 103 U/L; the activity of sucrase were declined from ( -0. 0107 ±0. 0003) × 103 U/L to ( -0.0078 ±0.0002) × 103 U/L; the△pH were declined from(2.8067 ±0.0404) to (2.5033 ±0. 0416) (24 h results). The differences were significant between experimental groups and the control group (P ＜ 0. 01 ), and there were also significant differences among experimental groups with different LPE concentration( P ＜0. 01 ). The inhibitory effect of acid generation and lactate dehydrogenas' activity of Sm were positively correlated ( P ＜ 0. 01 ). Conclusions LPE can inhibit the activity of lactate dehydrogenase,sucrase and the acid production capacity of the Sm in a dose dependent manner. The inhibitory effects in logarithmic phase is stronger than that in other phases of growth cycle.

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

Objective To investigate the clinical feature,treatment,and prognosis of hospitalized patients with jaw osteoradionecrosis. Methods A total of 93 cases with jaw osteoradionecrosis treated between 2000 and 2010 was reviewed.Of the 93 cases,79 cases were with mandible lesions,13 cases with maxillary lesions,and 1 case with both mandible and maxillary lesions.Sixty-six cases received one course of radiotherapy,with the radiation doses of 34 -90 Gy (mean 64.6 Gy).Twenty-two cases experienced tooth extraction or other operative procedures before exhibition of the clinical symptoms for osteoradionecrosis.The interval time between radiotherapy and the onset of osteoradioneerosis varied from 2 weeks to 32 years (mean 54 months).Results Of 93 cases,56 patients underwent radical resection of the pathologic bone and reconstruction with free tissue flaps,in whom 7 cases received the second surgery due to microvascular thrombosis in flap vessels,and flaps were survival by new vascular anastomosis in 3 cases,the failed flaps were removed and replaced successfully by non vascularized bone grafts in 2 cases,and the failed flaps removed and the defects were repaired with adjacent skin in other 2 cases.In the 56 cases,only one case was with disease recurrence and 53 cases with significant improvement in chewing and swallowing functions.Only 2 of 93 cases underwent resection of the pathologic bone and reconstruction with titanium plates,and thereafter they encountered titanium exposure.Scaling of osteoradionecrosis lesions was applied to 20 of 93 patients and 9 cases of them were with disease recurrence.Fifteen cases had resection of the effected mandible without reconstruction.Disease relapse was encountered in 2 of them,others had poor chewing and swallowing.Conclusions The mandible is more susceptible to osteoradionecrosis than maxilla.Radical resection with reconstruction by free tissue flap is recommended for the treatment of jaw osteoradionecrosis,and scaling and reconstruction only with titanium plate should be avoided because of high risks of titanium exposure and disease relapse.