Objective To construct adenovirus vector harboring CTLA4Ig-IRES2-I?B? gene for the study of enhancement blockade of T cell costimulatory pathway. Methods The IRES2 fragment was cloned in to the shuttle plasmid pAdtrack-CMV-CTLA4Ig, and then the human I?B? fragment was constructed with the link of the IRES2 to the pAdtrack-CMV-CTLA4Ig. Subsequently, adenovirus vector harboring CTLA4Ig-IRES2-I?B? was constructed by homologous recombination in E. coli BJ5183, and recombinant vector was packaged and propagated in 293 cells. Results The recombinant CTLA4Ig-IRES2-I?B? adenovirus was constructed by homologous recombination and identified by PCR and restrictive enzyme digestion methods. Conclusion The recombinant CTLA4Ig-IRES2-I?B? adenovirus may be used as a novel immunosuppressive agent for gene therapy in organ transplantation.

Objective To obtain the recombinant urease B subunit (ureB) of Helicobacter pylori (Hp) and apply it in serological detection of Hp infected patients. Methods Urease B subunit gene was amplified from the complete genome of Helicobacter pylori by PCR, and cloned into the PinPoint TM Xa Ⅲ fusion expression vector, then was sequenced. The protein of urease B subunit was expressed in E.coli JM109 and purified with affinity chromatography. 113 serum samples of peptic ulcer patients were detected by ELISA combined with purified UreB protein. Results DNA sequence analysis showed the nucleic acid sequence homology of ureB gene was 96.44% and the putative amino acid sequence homology was 99.65%. The recombinant UreB protein was composed of 571 amino acid residues and kept original immunologic reaction with corresponding antibody, and its purity was over 90% after affinity chromatography. The results of ELISA associated with recombination UreB antigen showed the sensitivity and specificity was 92%, 98.5%. Conclusion The recombinant UreB protein will be of value for clinical serodiagnosis and epidemiological study of Helicobacter pylori.

Objective To summarize the safety and efficacy of low dose of hepatitis B immunoglobulin (HBIG) for prevention of de novo hepatitis B infection after renal transplantation.MethodsThe clinical data of 138 patients who received renal transplantation without hepatitis B infection between January 2007 and June 2010 were retrospectively studied (study group).All the patients in study group were given low dose of HBIG injection before transplantation.The HBsAb titer was monitored regularly after transplantation,and the dosage of HBIG adjusted according to the level of the HBsAb titer.HBIG was implied to all patients in the study group for more than one year.The clinical data of 196 patients who received renal transplantation without hepatitis B infection between January 2004 and December 2006 served as the control group.These 196 patients were not treated with HBIG.The incidence of de novo hepatitis B infection,and acute rejection of these two groups was analyzed.The one-year graft and patients survival rate was also investigated.Results During the follow- up period of 12 months,only one case in the study group had de novo hepatitis B infection (0.7％) 6 months after renal transplantation,while 11 cases (5.6％) in the control group had de novo hepatitis B infection,in which 2 cases were died from acute hepatic failure.The incidence of de novo hepatitis B infection had statistically difference between the two groups (P＜0.05).The incidence of acute rejection in the study and control groups was 13.8％ and 17.3％ respectively (P＞0.05).The one-year graft and patient survival rate in the study and control groups was 96.4％ and 97.8％,and 90.3％ and 91.8％ respectively (P＜0.05).ConclusionLow dose of HBIG is effective and safe for prevention of de novo hepatitis B infection after renal transplantation.

Objective To evaluate the therapy effect of pneumatic lithotripsy treatment under ureteroscopic on urinary catheter encrustation(CE).Methods Eight patients with difficulty in pulling catheter tube crustcaused were enrolled as our subjects.They were performed pneumatic lithotripsy under ureteroscope operation.The information related to operation was recorded.Results Eight cases were successful in terms of crushing catheter encrustations,and the catheters were removed smoothly.Conclusion The therapy of pneumatic lithotripsy treatment under ureteroscopic is proved to be a minimally invasive treatment with small damage,and higher safety.

Objective To observe the effects of dexmedetomidine for different loading dose on blood pressure of surgery inten-sive care unit(SICU) patients conscious-sedation derivative phase .Methods 60 patients after general anaesthesia were divided into routine dose group(dexmedetomidine 1 .00 μg/kg ,group C) ,middle dose group(dexmedetomidine 0 .75 μg/kg ,group M ) and low dose group(dexmedetomidine 0 .50 μg/kg ,group L) ,and the loading time were reset 10 min .sedative effect was evaluated by Ram-say sedation score .Record the mean arterial blood pressure of everyone before administration ,5 min after ,15 min after ,30 min af-ter ,60 min after .Results All patients achieved good conscious-sedation .Compared with pro-administration ,the mean arterial blood pressure of group C descend significantly .The mean arterial blood pressure of group M and L descend significantly ,and then they were recover over time .60 min after the treatment ,the mean arterial blood pressure in group C was significantly lower than group L and goup M (P<0 .05) .Conclusion Small dose dexmedetomidine can achieved good conscious-sedation for SICU patients .It has less influence on hemodynamics .

Objective:To observe the effect of conversion from immediate-release tacrolimus (Tac) to extended-release Tac on kidney function in stable kidney transplant recipients.Methods:83 stable kidney transplant recipients who were converted from immediate-release Tac to extended-release Tac in the second people's hospital of Shanxi province from December 2011 to June 2019 were followed up for 12-36 months, and 83 stable kidney transplant recipients who continued to take immediate-release Tac were selected as control group.The changes of kidney function indexes, Tac trough concentration intra-patient variability (IPV) and compliance, the incidence of rejection and the survival rate of grafts and recipients were observed after the conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients.Results:The conversion time from immediate-release Tac to extended-release Tac was (42.76±30.50)months after transplantation. At 24 months after conversion, the serum creatinine (SCr) was significantly lower than that before conversion ( P=0.013), and the estimated glomerular filtration rate(eGFR)was significantly higher than that before conversion( P=0.005). In the experimental group , the SCr was significantly lower than that of the control group at 36 months after conversion ( P= 0.017), eGFR was significantly higher than that of the control group ( P=0.038). In the experimental group, the score of Immunosuppressant Therapy Barrier Scale (ITBS) was (20.23±2.89) before conversion and (17.63 ±3.08) after conversion ( P= 0.000). The daily dose of Tac was (2.09 ±0.84) mg before conversion and (2.10 ±0.83) mg after conversion. The trough concentration of Tac before conversion was (7.22 ±2.84) ng/mL, which reduced significantly after conversion. No rejection occurred after conversion, and the recipients/grafts survived healthily during the follow-up period. Conclusions:After conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients, the kidney function is stable and better than that of before conversion, the compliance is significantly improved, the IPV of Tac trough concentration is significantly reduced, and long-term use of extended-release Tac has good clinical efficacy and safety.

Objective To investigate the safety of ovarian preservation for stage Ⅰ endometrial carcinomas in women aged 40 years and younger.Methods Seventy-five cases of stage Ⅰ endometrial cancer aged 40 years and younger from Jan 1999 to Jan 2012 were treated in Cancer Hospital , Chinese Academy of Medical Sciences.They were further divided into two groups:20 patients who underwent ovarian preservation (group A) and 55 patients who underwent oophorectomy (group B).Clinical and pathological recordings of these patients were reviewed and compared.Results In the group A , there were 13 patients preserved both ovaries , and 7 patients preserved a single ovary.While there were no significant differences in the age, body mass index, surgical staging, histology, grade, cytology of peritoneal lavage or ascites , and postoperative treatment between two groups ( all P>0.05 ).The differences in the level of CA 125 [ 25%(5/20) versus 18%(10/55)] and number of patients underwent pelvic lymphadenectomy [35% (7/20) versus 84%(46/55)] were statistically significant between two groups (all P<0.05).Of seventy-five cases, only two patients relapsed and all survived after a median follow-up time of 31.7 months ( range:0 to 160 months).Kaplan-Meier analysis revealed no difference in overall survival (100.0% versus 100.0%) and disease free survival ( 90.0% versus 95.5%) between two groups ( P =0.579 ).Conclusions Ovarian preservation has no statistically significant impact on the survival of young patients with stage Ⅰa, well differentiated endometrial cancer.Large-scale, prospective clinical studies are needed to validate the safety of ovarian preservation for those patients.

AIM:To explore the recombined human proinsulin gene containing glucose reaction element(GLRE) expression in transfected CBRH7919 cells.METHODS: The packaged retrovirus encoding genetically modified human proinsulin PLXSN-(GLRE)3-BP-1MpINS3 and PLXSN-(GLRE)3-BP-1MpINS2 were transfected into CBRH7919 cells.Insulin values in cells after transient and steady expression screened by G418 at different glucose levels were detected.Chromosome DNA was isolated from transfected and untransfected cells and polymerase chain reaction(PCR) was performed.PCR products were analyzed by electrophoresis.RESULTS: 38 h after transfection,at the glucose levels of 0-25 mmol/L,the levels of insulin produced by cells including PLXSN-(GLRE)3-BP-1MpINS3 were(3.57?0.21)U/L,(5.30?0.20)U/L,(16.27?0.87)U/L,(23.23?1.12)U/L,respectively(P

Objective To explore the role of ERK1/2 in the central pathogenesis of migraine.Methods Healthy adult male SD rats were randomly divided into five groups:normal group (group C),sham operation group(group C),migraine model group(group M),DMSO group (group D)and PD-98059group (PD group),with 12 rats in each group.The extracellular discharge frequency in the spinal trigeminal nucleus was recorded and ERK1/2 phosphorylation was tested.Results (1) The percentage of extracellular discharge frequency change:Two hours after treatment,the percentage of discharge frequency change was (325.9±47.32)％.The percentage of extracellula discharge frequency change in group M (325.9±47.3)％ was higher than that in group N (100.0± 0.0) ％ and group C(107.3± 16.4)％.There was no significant difference in the percentage of discharge frequency change between group D(319.3±42.5) ％ and group M (325.9±47.3) ％.The percentage of discharge frequency change in group PD(218.5±31.7)％ was lower than that in group M(325.9±47.3)％ and group D(319.3± 42.5)％.(2) ERK1/2 phosphorylation:the ERK1/2 phosphorylation in group M and group D was higher than that in group N and group C.There was no significant difference in ERK1/2 phosphorylation between group D and group M.The ERK1/2 phosphorylation in group PD was lower than the other four groups.Conclusion During the process of central sensitization to migraine,neuronal excitability and ERK1/2 phosphorylation were increased.ERK1/2 inhibitor (PD98059) reduced ERK1/2 phosphorylation and neuronal excitability.These indicated that ERK1/2 may play a role in central sensitization of migraine in rats.

Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony-forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34+ cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU-GM was measured by using methylcellulose colony-forming assay, and the percentage of CD34+ and CD41a+ cells was measured by flow cytometry. Results: In control group，all the CD34+ cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34+ cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold，respectively at the third week while CFU-GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a+ cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions:HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a+ cells in direct contact coculture.