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.

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.

The fused gene (PTH-HSA) of parathyroid hormone (PTH) gene and Human Serum Albumin(HSA) gene was amplified without linker by Overlapping PCR technology. The spliced gene was clone into Pichia pastoris secretory vector pPIC9K. With the help of promoter AOX1 and mat ? signal peptide, the PTH-HSA gene was designed to secretory expression.Linearized by restriction enzyme SalI, The recombinant plasmid pPIC9K/PTH-HSA was transformed into Pichia pastoris KM71 by electroporation. The recombinant strains which were identified by G418 and PCR analysis were induced by methanol to express protein PTH-HSA. The target protein was expressed in fermentation supernatant. Western blot analysis of the fusion protein showed that the expressed fusion protein PTH-HSA had the antigenicity of HSA.adenylate cyclase assay proved that the fused protein exhibited the bioactivity to stimulate cAMP synthesis The specific activity of broth was about 318IU/ml.

This article introduces an overview of management system for conflict of interest of the European Pharmacopoeia Commission (EPC) and the United States Pharmacopoeia Convention (USP). The EPC and USP have standardized the management system for conflict of interest in drug standard work in multiple management documents, such as the Guide for the Work, Code of Practice for the Work, Form for Declaration of Interests and Confidentiality Undertaking of the EPC, bylaws, Rules and Procedures of the Council of Experts, Code of Ethics, Standards of Conduct of the USP, in order to ensure the transparency and fairness of drug standard development, improve the credibility and rigor of drug standards. This article introduces the management system for conflict of interest of the EPC and USP, providing reference for the improvement of relevant management systems of the Chinese Pharmacopoeia Commission.

Objective To assess the clinical factors impacting on the effective time of endocrine therapy for patients with prostate cancer.Methods The chnical data of 432 patients with prostate cancer who accepted endocrine therapy were analyzed retrospectively.The endpoint of the study was failure of endocrine therapy which was defined as continuous elevation of prostate specific antigen (PSA) from nadir for 2 times and more than 0.2 μg/L.The clinical data such as age,clinical stage,lymph node metastasis,bone metastasis,Gleason score,initial PSA,and PSA nadir were collected and their rehtionship with the effective time of endocrine therapy were further assessed via COX regression model.Results Age of onset was 57-88(73.70 ± 7.28) years.Initial PSA was 10.30-588.10(27.15 ± 75.90) μ g/L.The effective time of endocrine therapy was 3-62 (27.01 ± 13.10) months.Univariate regression analysis showed that initial PSA,clinical stage,Gleason score,PSA nadir,lymph node metastasis,bone metastasis were correlated with the effective time of endocrine therapy (P ＜ 0.01).Multivariate regression analysis showed that only Gleason score was correlated with the effective time of endocrine therapy(P=0.001).Compared with patients with Gleason score equal to or less than 3+4,patients with Gleason score equal to or more than 4+3 showed 2.49 fold increased risk of therapy failure (OR =2.49,95％ CI 1.44-4.30).Conclusion Gleason score has close relationship with the effective time of endocrine therapy for patients with prostate cancer,Gleason score equal to or more than 4+3 is an indicator for poor response to endocrine therapy.

Objective To compare the clinical efficacy between continuous and intermittent androgen deprivation in high risk prostate cancer.Methods Sixty-four patients with high risk prostate cancer were treated from January 2008 to April 2009,36 cases who accepted goserelin and bicalutamide were taken as intermittent hormonal therapy (intermittent treatment group),while 28 cases who accepted bilateral orchiectomy in addition to flutamide were regarded as continuous hormonal therapy (continuous treatment group).The comparison of tumor specific mortality,time of prostate specific antigen (PSA) to nadir,tine to PSA recurrence,serum testerone and quality of life score were assessed between the two groups.Results In continuous treatment group and intermittent treatment group,follow-up period was (26.4 ± 10.3) and (28.1 ± 8.7) months,the time of PSA to nadir was (3.8 ± 2.1 ) and (4.0 ± 3.6) months,the time to PSA recurrence was (20.1 ± 12.3) and (24.5 ± 14.6) months,respectively.There was no significant difference between the two groups.At the time of 18,24 and 30 months after therapy,serum testerone was 0.85,0.88,0.89 μg/L in continuous treatment group,while 1.21,1.36,1.48 μg/L in intermittent treatment group,respectively (P ＜ 0.05 ).Similarly,quality of life score was 38.7,40.5,39.8 scores in continuous treatment group,while 49.2,51.4,52.3 scores in intermittent treatment group at the time of 12,18 and 30 months after therapy,respectively (P ＜ 0.05 ).Conclusions Clinical efficacy could not been found between continuous and intermittent endocrinic therapy of prostate cancer.During intermittent,quality of life seems to be better and increases in accordance with serum testerone recurrence at given time.

OBJECTIVETo observe differences of therapeutic effects among acupuncture bloodletting, penicillin and acupuncture bloodletting combined with penicillin for children acute tonsillitis and providea better treatment method in cli nic.

METHODSSeventy-five mild cases were selected into section of mild symptoms while seventy-five severe cases were selected into section of severe symptoms. Cases in the two sections then were divided into, an acupuncture bloodletting group, a penicillin group and a comprehensive group by random digital table method separately, 25 cases in each one. Qu-chi (LI 11), Hegu (LI 4), Dazhui (GV 14), Shaoshang (LU 11) and Erjian (EX 11) were selected in the acupuncture bloodletting group, intravenous injection of penicillin sodium was applied in the penicillin group and acupuncture bloodletting combined with penicillin was applied in the comprehensive group. Efficacy assessment was conducted after 3 days in the section of mild symptoms and after 5 days in the section of severe symptoms.

RESULTSFor the section of mild symptoms, the total effective rate was 96.0% (24/25) in the comprehensive group and 92.0% (23/25) in the acupuncture bloodletting group, which were both superior to 68. 0% (17/25) in the penicillin group (P<0.05), but no statistical significance was seen between the comprehensive group and acupuncture bloodletting group (P>0.05). For the section of severe symptoms, the total effective rate was 96.0% (24/25) in the comprehensive group, which was obviously superior to 60.0% (15/25) in the acupuncture bloodletting group (P<0.01) and 68.0% (17/25) in the penicillin group (P<0. 05), and no statistical significance was seen between the acupuncture bloodletting group and penicillin group (P>0.05).

CONCLUSIONThe efficacy of acupuncture bloodletting combined with penicillin is little different from that of acupuncture bloodletting for treatment of children acute tonsillitis with mild accompanied symptoms, which were both superior to intravenous injection of penicillin sodium. For severe accompanied symptoms, the efficacy of acupuncture bloodletting combined with penicillin is obviously superior to acupuncture bloodletting and penicillin.

Acupuncture Points ; Acupuncture Therapy ; Bloodletting ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Humans ; Infant ; Male ; Penicillins ; therapeutic use ; Tonsillitis ; drug therapy ; therapy ; Treatment Outcome

A series of isoindoline derivatives were designed, synthesized, and evaluated for their double inhibitory activities. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Preliminary in vitro pharmacological tests showed that all compounds exhibited 5-HT or NE reuptake inhibition activity. Among the tested compounds, compound I-3 exhibited potent inhibitory activity against 5-HT and NE reuptake in vitro, and exhibited potent antidepressant activity in vivo. These compounds designed can be further optimized for finding more potent 5-HT/NE dual reuptake inhibitors and antidepressant candidates as well.
Antidepressive Agents ; chemical synthesis ; chemistry ; Biological Transport ; Drug Design ; Isoindoles ; chemical synthesis ; chemistry ; Serotonin Uptake Inhibitors ; chemical synthesis ; chemistry ; Structure-Activity Relationship

Arterio-Arterial Fistula ; congenital ; therapy ; Coronary Artery Disease ; congenital ; therapy ; Follow-Up Studies ; Heart Defects, Congenital ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome