Objective To prepare the monoclonal antibody against human perforin(HP). Methods Recombinant eukaryotic expression plasmid pCDM8-HP was extracted and purified, and the BALB/C mice were immunized with the plasmid. The hybridomas producing anti-HP McAbs were established by using hybridoma technique, then the specificity of the McAbs was identified by using immunocytochemical technique and Western blot. Results Three hybridoma cell lines secreting McAbs against human perforin were established, and the three McAbs showed positive only with LAK cells containing human perforin protein, and showed negative with inactive human peripheral blood lymphocytes (PBLs). The subclasses of the three McAbs were determined as lgG2bK. Western blot results showed that the three McAbs recognized a specific band of LAK cell lysateds with molecular weight of 70.0Kd. Conclusion The three hybridoma cell lines secreting McAbs against human perforin were established and the secreted McAbs were specific.

Objective To investigate effect of serum apoptosis factor and long-term effect on medical abortion patients by sequential misoprostol. Methods 80 cases of early pregnancy patients who required termination pregnancy, were selected and randomly divided into 2 groups.40 cases in control group were treated with mifepristone 25mg oral, misoprostol 600μg, 4 times a day.40 cases in experiment group were treated with sequential use of misoprostol.Effect of medical abortion, survivin and caspase-3 were compared after 7d treatment.Gynecological inflammation, dysmenorrhea and infertility were compared for 2 years.Results Compared with control group, complete abortion rate of the experiment group was higher (P<0.05).The incidence of adverse drug reactions of the experiment group was lower than control group (P<0.05) after 2 years follow.up.Compared with the control group, HCG,E3 and P of experimental group were lower (P<0.05).After 7 days of medication,survivin decreased,caspase-3 increased,compared with control group, survivin of the experiment group was lower, caspase-3 was higher ( P<0.05 ) .Conclusion Misoprostol sequential can improve the success rate of medical abortion abortion, reduce the complications of abortion, presumably with the inhibition of survivin expression, is associated up regulation of caspase-3 levels.

Objective To express human B7.2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-α. The fusion protein consisted of GST and hB7.2(IgV+C) was identified by SDS-PAGE and Western blotting.T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated by anti-CD3 antibody. Results The fusion protein GST-hB7.2 (IgV+C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7.2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.

Objective To explore the impact of sleep quality in early pregnancy on gestational diabetes. Methods A total of 657 pregnant women in the obstetrics clinic of our hospital were selected as the research subjects. The basic information of pregnant women, sleep time and sleep quality in early period of pregnancy were collected through questionnaires. Glucose tolerance screening tests were performed at 24-28 gestational weeks to screen out GDM pregnant women. Multivariate unconditional logistic regression analysis was used to analyze the effect of sleep status in early pregnancy on the incidence of gestational diabetes. Results 112 pregnant women were diagnosed with gestational diabetes, and 92 women in early pregnancy were indicated of abnormal sleep quality. Sleep disorders might lead to an increased risk of gestational diabetes (OR=1.031, 95％CI=1.027-1.115). Age, pre-pregnancy BMI, early pregnancy sleep quality and number of pregnancy and delivery were risk factors for gestational diabetes. Conclusion Poor sleep quality in early pregnancy is a high risk factor for gestational diabetes. Treatment of abnormal sleep during early pregnancy can reduce the incidence of gestational diabetes.

OBJECTIVETo investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro.

METHODSThree fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation.

RESULTSThree recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC.

CONCLUSIONSFunctional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.

Antigens, CD ; pharmacology ; B7-2 Antigen ; Escherichia coli ; genetics ; Humans ; Immunoglobulin Constant Regions ; pharmacology ; Immunoglobulin Variable Region ; pharmacology ; Lymphocyte Activation ; Membrane Glycoproteins ; pharmacology ; Plasmids ; Recombinant Fusion Proteins ; pharmacology ; T-Lymphocytes ; immunology