Objective To observe apoptosis of primary cultured synoviocytes from patients with rheumtoid arthritis (RA) which induced by transfected recombinant plasmid carring Fas ligand (FasL) gene, aimed to detect the aim targets and develope gene therapy for RA by intra-articular injection. Methods FasL cDNA was introducted into the plasmid pcDNA3.1-neo by reverse transcriptase-polymerse chain reaction (RT-PCR) to set up the recombinated human FasL gene expression carrier, Then the pcDNA3.1-FasL was transducted into the primary cultured synovial cells by catholyte liposomes. After screened by G418, transgenic synovial cells were choosen. Finally through the microscope, TUNEL, Annexin Ⅴ-FITC and PI staining with flow cytometry, we observed the apoptosis state of each synovial cell. Results Prokaryotic and eukaryon carrier with FasL gene were established successfilly. The gene sequencing had completely coincident with the lierature. When eukaryon carrier with FasL (pcDNA3.1-FasL) was transducted into the index number livinging RA synovial cells after 15 h, most synovial cells presented distortion transformation, and the minority sheded from the bottom wall. The transgenic RA synovial cells after screened by G418, only a little amount survived about 1~2 per sight, and the cells grew slowly, the normal matched cells were cloned after 2~4 weeks. The FasL gene transgenic RA synovial cells presented positive findings by TUNEL staining. Early apoptosis cells by Annexin Ⅴ/PI staining, but the difference wasn′t significant. Conclusion The eukaryon carrier with FasL (pcDNA3.1-FasL) can be transferred into RA synovial cells. It can induce synovial cell apoptosis.

A total of 183 patients of ectopic pregnancy due to tubal factors were divided into the methotrexate (MTX),conservative surgery and salpingectomy groups.The dose of gonadotropin,counts of harvested oocytes and high-quality embryos and pregnancy rate were compared among three groups to analyze the differences of in vitro fertilization and embryo transfer (IVF-ET).And the above parameters showed no significant differences (P ＞0.05 ).The clinical pregnancy rate of the conservative group was higher than the other two groups.And the difference was statistically significant compared with salpingectomy ( P ＜ 0.05 ).It suggested that the treatments of ectopic pregnancy had some effects on the outcome of IVF-ET.The pregnancy rate was slightly higher in the conservative surgery group.

Objective:To elucidate the role of ERK1/2 MAPKs signal transduction in the protective effects of ischemic preconditioning during liver transplantation. Methods: Twenty-four rats were equally randomized into 4 groups: sham control (group A); liver transplantation (group B); ischemic preconditioning+liver transplantation (group C), and MEK inhibitor intervention+ischemic preconditioning+liver transplantation (group D). Serum AST and ALT were detected after operation and cellular ultrastructures were observed by transmission electron microscopy. Liver tissue ERK1/2 MAPKs phosphorylation activities were evaluated through determining ERK-1 protein phosphorylation by Western blotting. Results: Serum ALT and AST activity in group B and group D was significantly higher than those in group A, and remained normal in group C. Liver tissue ERK1/2 MAPKs phosphorylation activity increased significantly in group C, and the activation effect was inhibited in group B and D. Cellular ultrastructure was obviously damaged in group B and group D, and the damage was prevented in group C.Conclusion: Ischemic preconditioning has a protective effect on transplanted liver cells in which ERK1/2 MAPKs pathway plays a pivotal role.

AIM: To investigate the effects of alanyl-glutamine (Ala-Gln) on expression of iNOS and TNF-? in injured intestinal mucosa induced by oral tacrolimus(FK506). METHODS: Twenty-four BALB/c mice were randomized to receive orally 0.2 mL of normal saline solution ( groupⅠ), 0.2 mL of FK506 in a dose of 0.1 mg/kg (groupⅡ) or 1.0 mg/kg (groupⅢ), and orally high-dose FK506 (0.2 mL, 1.0 mg/kg) plus intraperitoneal injection of Ala-Gln (0.5 g/kg )(groupⅣ),respectively. Damages of intestinal mucosa were determined by pathological examination. Intestinal mucosal permeability was analysed by FITC-dextran fluorescence assay. Expression of iNOS and TNF-? in intestine was detected by RT-PCR and Western blotting.RESULTS: Severe damage on the villi and increased intestinal permeability were observed in high-dose FK506 treated mice according to scanning electron microscopy and FITC-dextran flux respectively. The erosion and increased intestinal permeability were significantly alleviated by Ala-Gln treatment. Transcription of iNOS mRNA and TNF-? mRNA, which was up-regulated in high-dose FK506 treated group, was also markedly down-regulated in mice combined with Ala-Gln-treatment. A significantly increased expression of iNOS and TNF-? protein was found in the high-dose FK506 treated mice, while small amounts of these proteins were identified in the Ala-Gln-treated group.CONCLUSION: FK506 could induce a significant impairment of intestinal mucosa morphologically, which might be associated with up-regulated expression of iNOS and TNF-? in small intestinal mucosa. Subsequently, the intestinal permeability is increased. Ala-Gln has a strong protective effect on FK506-induced intestinal barrier dysfunction, probably relates to the down-regulation of iNOS and TNF-? expression.

BACKGROUND: Considerable debate exists which factors influence the outcomes of the vitrified frozen-thawed embryo transfer because lack of clinical applications.OBJECTIVE: To explore the factors influencing the outcomes of the vitrified frozen-thawed embryo transfer in assisted reproductive technology.METHODS: A retrospective statistical analysis was performed in Reproductive Medicine Center, Renmin Hospital of Wuhan University of 142 patients, 154 thawing cycles. The patients were grouped according to the age of patients, embryonic development, fertilization methods, endometrial preparation programs, the endometrial thickness, the process of transplantation and the survival cell ratio of embryo recovery, the implantation rate and clinical pregnancy rate were compared between various groups.RESULTS AND CONCLUSION: Among the groups of the different age, fertilization methods, endometrial preparation programs, endometrial thickness and the process of embryo transfer embryo implantation rate and clinical pregnancy rate were no significant difference (P > 0.05); between the two groups of the second day fertilized embryos (D2) and the third day (D3)fertilized embryos, the clinical pregnancy rate was not significant (P> 0.05), but the embryo implantation rate of D3 group was significantly higher than D2 group. The survival cell ratio of embryo recovery has a significant effect on implantation rate and clinical pregnancy rate (P < 0.05). Ln the frozen-thawed embryo transfer cycles, embryo quality plays a major role in pregnancy rate, and preparation for appropriate endometrial thickness can improve the clinical pregnancy rate.

Objective To investigate the efficacy and safety as well as the effects of rituximab on B-lymphocytes and anti-platelet glycoprotein-specific antibodies,in patients with steroid-resistant idiopathic thrombocytopenic purpura(ITP).Methods Twelve steroid-resistant ITP patients,16 to 54 years old,received intravenous rituximab at the dose of 375 mg/m2 once-weekly for 4 weeks.Lab studies included CBC,serum concentrations of IgG,IgM and IgA.CD+3,CD+4,CD+8,CD+19,CD+20 cell numbers were assayed by flow cytometry and anti-platelet glycoprotein-specific antibodies(GP Ⅱ b/Ⅲ a,GP Ⅰ b/Ⅸ)were assayed by monoclonal antibody-specific immobilisation of platelet antigens prior to and following rituximab therapy.Results A complete response(platelet counts ≥100×109/L)was observed in 4 cases,a partial response (platelet counts between 50 and 100×109/L)in 3 cases,a minor response(platelet counts between 30 and 50×109/L)in 2 cases,and non response(platelet counts＜30×109/L)in 3 cases.Responses were sustained 0.5 to 12 months(median 5 months).After 4 weeks of rituximab therapy,anti-platelet glycoprotein-specific antibodies(GP Ⅱ b/Ⅲ a,GP Ⅰ b/Ⅸ)disappeared except one NR patient and CD+19/CD+20 cells were almost depleted in all patients(295.0±86.4)×106/L vs(4.1±2.2)×106/L(P＜0.01).As expected,the T cell counts,and the serum concentrations of IgG,IgM and IgA were not changed after therapy.No severe side effects were observed.Conclusion Rituximab may be an effective and safe treatment for adults with steroid-resistant ITP.

AIM: To investigate the effects of alanyl - glutamine ( Ala -Gln) on expression of iNOS and TNF- α in injured intestinal mucosa induced by oral tacrolimus (FK506). METHODS: Twenty -four BALB/c mice were randomized to receive orally 0.2 mL of normal saline solution ( group Ⅰ ), 0.2 mL of FK506 in a dose of 0.1 mg/kg ( group Ⅱ ) or 1.0 mg/kg (group Ⅲ), and orally high -dose FK506 (0.2 mL, 1.0 mg/kg) plus intraperitoneal injection of Ala -Gln (0.5 g/kg )(group Ⅳ ),respectively. Damages of intestinal mucosa were determined by pathological examination.Intestinal mucosal permeability was analysed by FITC - dextran fluorescence assay. Expression of iNOS and TNF - α in intestine was detected by RT - PCR and Western blotting. RESULTS: Severe damage on the villi and increased intestinal permeability were observed in high - dose FK506 treated mice according to scanning electron microscopy and FITC - dextran flux respectively. The erosion and increased intestinal permeability were significantly alleviated by Ala - Gln treatment. Transcription of iNOS mRNA and TNF - α mRNA, which was up - regulated in high - dose FK506 treated group,was also markedly down- regulated in mice combined with Ala- Gln- treatment. A significantly increased expression of iNOS and TNF - α protein was found in the high - dose FK506 treated mice, while small amounts of these proteins were identified in the Ala - Gln - treated group. CONCLUSION: FK506 could induce a significant impairment of intestinal mucosa morphologically, which might be associated with up - regulated expression of iNOS and TNF - α in small intestinal mucosa. Subsequently, the intestinal permeability is increased. Ala - Gln has a strong protective effect on FK506 - induced intestinal barrier dysfunction, probably relates to the down - regulation of iNOS and TNF - α expression.

Objective To study the effect of recombinant human endostatin (rhES)on the radiosensitivity of esophageal squamous cells KYSE-150 and its preliminary mechanism.Methods Cells were divided into four groups:control group without treatment,rhES group treated with recombinant human endostatin,radiation alone group exposed with X-rays,and combination group exposed with X-rays plus endostatin.Colony formation assay was used to measure cell survival fraction.A single-hit multi-target model was used to fit cell survival curve and calculate the sensitive enhancement ratio (SER).Influence of rhES combined with X-ray radiation on cell cycle and apoptosis was measured by flow cytometry.Expressions of Cyclin B1,Cyclin D1,Bcl-2 and Bax mRNAs were determined by RT-PCR.Protein expressions of HIF-1α,VEGF,and VEGFR were determined by Western blot.Results D0,Dq and SF2 value of KYSE-150 cells decreased along with the concentration of rhES.At D0dose,the SER for 100 and 200 μg/ml rhES was 1.14 and 1.27,respectively.Compared with the radiation alone group,the apoptosis rate and bax expression increased,while the expressions of VEGF and HIF-1α decreased in the combination group (t =7.97,3.02,117.55,7.22,P ＜ 0.05).Conclusions rhES has radiosensitive effect on esophageal carcinoma cells KYSE-150 in vitro by inhibiting the expressions of HIF-1α and VEGF,regulating bax expression,and inducing apoptosis.

Objective To explore the feasibility of laparoscopic-assisted pancreaticoduodenectomy.Methods The clinical data of 5 patients in our hospital from January to May 2010 were analyzed. 2 patients were pre-operatively diagnosed to have lower common bile duct adenocarcinoma, and 2 patients were preoperatively diagnosed to have adenocarcinoma of the descending duodenum, 1 patient was intra-oparatively diagnosed to have pancreatic head cancer. During the operation, laparoscopic exploration was performed, then gallbladder, distant bile duct, distant stomach, duodenum, part of jejunum and head of pancreas were disassociated, then the digestive tract was reconstructed under open abdomen surgery. Results All the operations of the 5 cases were successfully performed, with an average operation time ( 339 ± 54) min and an intra-operative blood loss of (538 ± 106)ml, and there was no intra-operative blood transfusion. The patients'bowel function recovered (4.0 ± 1.0 ) d postoperatively and were discharged ( 15.8 ± 4.7 ) d postoperatively.1 patient developed pancreatic fistula and was cured with conservative treatment. Conclusions Laparoscopicassisted pancreatoduodenectomy is minimally invasive with short operation time and fast postoperative recovery,which is worth of further clinical study.

Objective To investigate the association of tumor necrosis factor (TNF)-α, heat shock protein (HSP)70-2 gene polymorphisms and susceptibility of acute pancreatitis(AP). Methods Using case-control method,The gene polymor?phism of TNF-α and HSP70-2 was detected by PCR-RLFP in 72 patients with AP and 71 healthy controls. Results There were no significant differences in proportion of TNF-αgenotype and alleles between AP and control groups (P>0.05). There were no significant differences in TNF-αgenotype and alleles between severe acute pancreatitis (SAP) and light acute pancreatitis (MAP) of AP group (P>0.05). There were no significant differences in white blood cell count, C-reactive pro?tein (CRP), amylase, three acyl glycerin and glucose between TNF-a and HSP70-2 gene of AA type and GA+GG type pa?tients (P>0.05). The HSP70-2 genotype GA+GG proportion was significantly higher in AP group than that of control group (69.4%vs 49.3%). The ratio of patients with G allele was significantly higher in AP group than that of control group(46.5%vs 31.7%). The ratio of patients with GA+GG type AP was significantly higher in SAP patients than that of MAP patients of AP group(81.0% vs 53.3%). There was no significant difference in G allele between SAP and MAP patients (P>0.05). Conclusion TNF-α polymorphisms is not associated with acute pancreatitis. There is an association between HSP70-2 polymorphisms and acute pancreatitis. Carrying the G allele increases the possibility of a severe acute pancreatitis ,which is one of the genetic susceptibility factors of severe acute pancreatitis.