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.

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

Objective To review our patients who underwent reconstruction of the defect after combined cranio-maxillo-facial resection in recent years.Methods From January,2005 to January,2011,88 patients underwent reconstruction of the defect after combined cranio-maxillo-facial resection.Different reconstructive techniques were used according to the defect classifications in dura,skull base bone,and facial tissues.For dural defects,no repair (37 cases),primary closure (25 cases),and dural repair (26 cases) were performed,respectively.Dural repair materials included thigh fascia lata (2 cases),temporalis fascia (2 cases),pericranium (1 case) and artificial dural patch (21 cases).Bone reconstruction of the skull base were performed in 61 patients with titanium mesh (57 cases),free iliac bone graft (1 case),free cranial bone graft (2 cases) and Medpor (1 case),respectively.Limited facial soft and hard tissue defects in 44 patients were reconstructed with temporalis system of flaps (36 cases),sternocleidomastoid myocutaneous flap (6 cases),and pterygoid muscle flap (2 cases),respectively.Extensive facial soft and hard tissue defects in 44 patients were reconstructed with free latissimus dorsi myocutaneous flap (26 cases),free pectoralis major myocutaneous flap (12 cases),free anterolateral thigh perforator flap (5 cases cases) and free anteromedial thigh perforator flap (1 case),respectively.Results The overall success rate of 88 flaps was 100％.Cerebrospinal fluid leak was found in 4 patients,wound infection was found in 2 patients,intracranial infection was found in 2 patients,respectively.Six patients with cerebrospinal fluid leak or wound infection were cured by conservative treatment.Two patients with intracranial infection were dead although they underwent salvage surgery.Overall rate of complications was 9.1％,dead rate was 2.3％.Conclusion Successful reconstruction of the defect after combined cranio-maxillo-facial resection can be achieved by watertight dural repair,bone reconstruction of the skull base and well-vascularized tissue covered.Regional flap and free tissue transfer are both preferred reconstructive technique depending on the anatomic site and the extent of the defect.

Objective To study osteoporosis in patients receiving androgen deprivation therapy (ADT) with prostate cancer, and determine whether once-weekly oral alendronate can prevent bone loss in men receiving ADT. Methods One hundred and twelve men with nonmetastatic prostate cancer receiving ADT were divided into two groups from April 2007 to April 2008, 56 cases in each group. Group A took alendronate (70 rng once-weekly orally) and calcium supplement, group B received calcium supplement only. Bone mineral density (BMD) were measured both before and 6 months, 12 months after treatment for both groups. Results There were no statistically differences in age, persistence time of castration, prostate specific antigen level and adverse effect between two groups(P＞ 0.05). At baseline, 39.3%(44/112) of men had osteoporosis and 51.8%(58/112) had low bone mass. After 12 months treatment, in group A, BMD increased 3.7% (95% CI 2.80% to 4.60% ,P＜0.01 ) at the spine,0.7%(95% CI 0.10% to 1.40% ,P=0.031)at the total hip and 1.6% (95% CI0.40% to 2.80%,P =0.008) at the femoral neck. In group B decreased 1.4% (95% CI-2.70% to -0.03%, P = 0.045 ) at the spine, 0.7% (95% CI - 1.50% to -0.01%,P = 0.052) at the total hip and 0.7% (95% CI -1.50% to 0.10%, P = 0.081 ) at the femoral neck. The estimated changes in BMD were significantly different between two groups (P ＜ 0.01 ). Conclusions It suggests that ADT induce bone loss which should be treated in early stage. Bone loss that occurred with ADT is prevented and improved with once-weekly oral alendronate.

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