Objective:To produce a fusion protein(ICOS-Ig)between extracellular portion of human ICOS and Fc portion of mouse IgG 2a and study on its biological activity.Methods:cDNA encoding the extracellular domain of human ICOS was prepared by RT-PCR from the RNA of the stimulated human peripherial blood mononuclear cells. The Fc portion of mouse IgG2a was cloned by PCR from the vector that contains the sequence-encoding Fc portion of mouse IgG2a.Two resulting amplified PCR products were ligated into a secrection mammalian expression vector, pSecTag2/Hygro A. The recombined vector was transfected into CHO cells by lipofectamine2000.Results:RT-PCR demonstrated the integration and mRNA synthesis of fusion gene. ELISA and Western blot analysis showed the expression of ICOS-Ig and its molecular weight was between 43-66 kD, its concentration was 5-25 ?g/ml. FACS analysis assured that ICOS-Ig had ligand specific binding activity. Addition of ICOS-Ig to MLR resulted in inhibition of T-cell proliferation and IL-2,IFN-? secretion.Conclusion:The fused gene ICOS-Ig was constructed and expressed successfully. It had the bioactivity of inhibition of T cell proliferation and cytokine secretion.

Objective To evaluate the clinical application of HLA matching in highly sensitized recipients of renal allografts.Methods Recipient's panel reactive antibody (PRA) was detected by using ELISA test with Lambda antigen tray (LAT).Donor and recipient HLA classⅠtyping was performed with special monoclonal tray,and HLA classⅡgene typing with micro-sequence specific primers (Micro-SSP).Results There were 104 recipients with anti-HLA class-ⅠIgG antibody,76 with anti-HLA class-ⅡIgG antibody,and 44 with both anti-HLA class-1 and anti HLA class-ⅡIgG antibody respectively in 136 sensitized recipients.HLA class-ⅠIgG antibody positive rate was 11%-97 %,with an average of 49.6%?23.8%;The common public epitopes antibody was not found in each recipient of 13 cases with PRA＜20%,but was found in I2 recipients in 44 cases with PRA be- tween 20%-50%,and 39 recipients in 47 cases with PRA＞50%.HLA class-ⅡIgG antibody posi- tive rate was 17%-100%,with an average of 28.2%?63.8%.The number of cases of 0,1,2,3, 4 MM was 7 (5.1%),26 (19.1%),47 (34.6%),39 (28.7%) and 17 (12.5%) respectively by the standard of conventional HLA antigen matching;however the number of the recipients with 0,1, 2,3 MM was 31 (22.8%),53 (39.0%),36 (26.5%) and 16 (11.7%) respectively according to the rule of HLA CREGs matching and none with 4 MM.Rates of acute rejection in sensitized recipi- ents with 2MM and 3MM HLA-CREGs were 25.0% and 37.5% respectively and were significantly higher than those with 0MM (P＜0.05,＜0.05 respectively).Kidney year-survival was decreased when the number of MM of HLA CREGs matching increased.Conclusion The HLA CREGs matching can improve the ratio of well-matched significantly.Good HLA matching can reduce the incidence of acute rejection in sensitized recipients and increase the survival rate of grafts.

Adult ; Arterio-Arterial Fistula ; therapy ; Cardiac Catheterization ; Coronary Vessels ; pathology ; Heart Ventricles ; pathology ; Humans ; Male

Objective:To study the bioactivity of inducible costimulator Ig(ICOS-Ig) as an inhibitor of ICOS-B7RP-1 costimulatory pathway in vitro. Methods: cDNA encoding the extracellular domain of human ICOS was prepared by RT-PCR from the RNA of the stimulated human peripheral blood mononuclear cells. The Fc portion of mouse IgG2a was cloned by PCR from the vector containing the sequence-encoding Fc portion of mouse IgG2a.The above 2 PCR products were ligated into a clone vector: pGL-3-Basic. The fusion gene was then cloned and ligated into a mammalian expression vector: pcDNA4/HisMAX A. The recombined vector was transfected into CHO cells by Lipofectamine2000 and the expression of the fusion protein was identified by Western blot. The mixture lymphoproliferation reaction(MLR) of the lymphocytes derived from BALB/c and C57BL mice was used to detect the fusion protein function in vitro. Results: Western blot analysis showed the expression of fusion protein, with the molecular weight being 43 000-66 000. FACS analysis assured that expression products had ligand specific binding activity. MLR was inhibited by the fusion protein. Conclusion: The constructed recombinant fusion protein has ligand specific binding activity and can inhibit the lymphoproliferation.

Objective To observe the clinical efficacy and safety of Shenxiong glucose injection and lipo PGE1 injection in treating the lower extremity arteriosclerotic occlusive disease (LEAOD).Methods 80 patients with LEAOD were randomly divided into control group and treatment group. Both groups were given conventional therapy, including reducing blood pressure, blood sugar, blood lipid and anti-infection therapy.Control group was additonally given intra-femoral arterial infusion with urokinase 150000 units plus 15mL 0.9% sodium chloride and 10 mg anisodamine alternately every other day, 5 days for one course, stopping 3 days after another course.Treatment group was treated with intravenous injection Lipo PGE1 injection plus 10 mL 0.9% sodium chloride and intravenous Shenxiong glucose injection per day for 14 days.Clinical efficacy, changes in clinical symptoms, whole blood viscosity, high sensitivity C-reactive protein(hs-CRP), fibrinogen, ankle-brachial index(ABI), the inner diameter of the dorsalis pedis and the peak systolic velocity( PSV) of dorsalis pedis were compared and analyzed pre and post-treatment.Adverse drug reactions were recorded during the treatment.ResuIts The efficiency for the patients in the treatment group (90.0%) was higher than that in control groups(72.5%) (P<0.05).The symptoms of numbness and cold limbs, whole blood viscosity, ABI, hs-CRP improved more significantly in the treatment group (P<0.05).No adverse event occurred in the treatment group and 2 patients in control group had mild dry mouth.ConcIusion Shenxiong glucose injection combined with lipo PGE1 injection for the treatment of LEAOD is effective and safe and should be introduced in clinical practice.

Objective To understand the sleeping time and the prevalence of sleep disturbances in children aged 3-12 years in Wuxi city.Methods Two thousand and three hundred seventy six children aged 3-12 years were investigated with questionnaires from June to September 2003.Results The average sleep time of 1 day in each age group was 11.61,10.91,10.68,10.27,9.81, 9.67,9.61,9.57,9.60,9.61 hours, respectively. The total prevalence of sleep disturbance was 25.5%. Among them, the sleep snoring prevalence was 7.8%; choke/gargling was 0.63%;mouth breathing was 6.6%;sleep apnea was 0.34%;rubbing teeth was 7.7%;sleep talking was 4.2%;somnambulate was 0.63%;enuresis was 1.4%;limber spasm was 3.2%;sleep inquietude was 6.2%。Conclusion Prevalence of sleep disturbance in children age 3-12 years was high and sleep time was shorter in Wuxi than that in other cities.

BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility.OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell (BMSC) homing and the therapeutic efficiency on ischemic stroke. METHODS: A middle cerebral artery occlusion (MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group (n=15), BMSCs group (n=18), ultrasound+BMSCs group (n=18), ultrasound+microbubble+BMSCs group (n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×106 BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation (1 MHz, 2 W/cm2) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group.RESULTS AND CONCLUSION: (1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups (P < 0.05). (4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups (P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke.

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Objective To investigate the feasibility of treatment mode of end-to-end anastomosis of esophagus(EAS) af ter partial resection for early-stage cervical esophageal carcinoma(ECEA).Methods 7 patients were substantially confirmed as squsmous cell carcinoma of cervical esophagus by endoscopy,the nearest distance of the lesion from the incisors was 17cm,and the furthest was 20 cm,the maximum extent was 2.5 cm,and the minimum was 1 cm.None of them with longitudinal muscularis invasion.Confirmed by PET/CT or chest enhancement CT examination preoperatively,intrathoracic and cervical lymphatic metastasis was excluded,cT1 -2 N0 M0.Incisal margin length was not less than 1 cm,the maximum was 5 cm and the minimum was 3 cm.Meanwhile,the cervical lymph node should be dissected,and the average number was 6.43 per case.After surgery,all the patients were fixed by plaster slab to release the tension of anastomosis.Postoperative adjuvant radiotherapy or chemotherapy was received.Results None of the patients had severe postoperative complications,and the average hospital stay was 14.5 days.All the patients are alive,the longest follow-up lasts for 3 years and 4 months,all of them can take normal food,without anastomotic stenosis.Conclusion Treatment mode of EAS after partial resection for ECEA significantly decrease the operative damage,apparently improve the patient's quality of life(QOL),so that the patients can better receive adjuvant treatment subsequently; it is a feasible and effective method for cervical esophageal carcinoma at the early stage.

Objective To explore the relationship of post-transplant major histocompatibility complex class I chain-related gene A(MICA)antibody status and renal allograft function in clinical stable phase.Methods Fifty-seven patients accepted renal allografts followed up for at least 6 months were detected with the levels and specialties of MICA antibodies by Flow PRATM beads.Simultaneously,their serum ereatinine levels were tested as well.The impact of MICA antibody status on renal allograft function was assessed.Results Among the 57 patients,38 cases showed no HLA and MICA antibody.11 cases had HLA antibodies but not MICA antibody,8 cases had MICA antibodies and 3 cases had both MICA and HLA antibodies.There were 5 patients with MICA019 antibodies.3 patients with MICA027 antibodies,2 patients with MICA018 antibodies,while 1 patient with MICA004 and MICA017 antibodies,respectively.There were 9 patients with antibody positive score higher than 6,accounting 75%(9/12).Except age,there was no significant difference between patients with positive and negative MICA antibodies in the aspects of blood transfusion history,CDC,and cold ischemia time(P＞0.05).The average ages were(32.5±7.9)years for MICA antibodypositive patients and were(43.0±1 0.4)years for MICA antibody-negative patients(P=0.008).MICA antibody-positive patients without HLA antibody had higher serum creatinine level[(117.20±12.30)μmol/L]than MICA and HLA antibody-negative patients[(89.40±28.95)μmol/L,P＜0.05].Conclusions The measurement of MICA antibodies has prognostic value in the assessment of patients without HLA antibodies after renal transplantation.MICA antibody positive has clear association with chronic renal allograft function decline.