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

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.

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.

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

The hom gene encoding for homoserine dehydrogenase was amplified from the genomic DNA of Corynebacterium glutamicum ATCC 13032.After the kanamycin-resistant gene(Km)cassette from plasmid pET28a was inserted into the center of hom,the hom::Km cassette was then electroporated into the competent cell of C.glutamicum ATCC 13032.And kanamycin-resistant clones were obtained.PCR was performed to confirm whether the Km gene was integrated into the hom gene of these clones and the recombinant strains of hom-disrupted were screened out.Fermentation results showed that the lysine yield of the hom-disrupted strain C.g-hom::Km-8 reached 4.7 g/L,which was 6.7 times that of C.glutamicum ATCC 13032.

Objective To evaluate the efficacy and safety of ESHAP regimen,as a salvage regimen, in treating patients with relapsed or refractory aggressive NHL.Methods 38 patients with relapsed or refrac- tory aggressive NHL were selected to be treated by ESHAP regimen.Results The 38 patients received ES- HAP regimen with a range of 2～6 cycles. The total RR was 55.3 % with complete response(CR)rate of 26.3 %.The major toxicity was myelosuppression with infection,which was tolerable.Conclusion ESHAP regimen is one of safe and effective salvage regimens for the patients with relapsed or refractory aggressive NHL.

Objective To investigate MR imaging features of femoral marrow in treated ?-thalassemia major.Methods MR imaging of the proximal femoral marrow was performed in 35 cases of ?-thalassemia major and 45 age-and sex-matched normal children as control.Coronal images of femoral marrow with the techniques of spin echo and fast field echo(FFE)were obtained.On T_1-weighted imaging the red and yellow femoral marrow were judged and marrow distribution was classified into five groups.The hemosiderosis of marrow was judged on the basis of signal intensity of marrow on FFE imaging.The marrow distribution classification and the hemosiderosis on MR imaging were correlated with clinical features.Results On FFE,marrow hemosiderosis occurred in 15 patients with a marked hypo-intensity signal and was related to the age(P=0.032).On T_1-weighted imaging,the femoral marrow in 35 patients was classified as groupⅢand IV,while the marrow distribution was groupⅠorⅡin all normal children,there was statistically significant difference(P