Safety, tolerability and pharmacokinetic study of recombinant human parathyroid hormone in healthy male Chinese subjects

AIM： To determine the safety, tolerability and pharmacokinetic parameters of a new drug recombinant human parathyroid hormone [ rhPTH （1-84）] in healthy male Chinese subjects. METHODS： domly divided Thirty-six healthy male volunteers were rangroups received into 3 groups. The volunteers in these single subcutaneous injection of rhPTH （ 1-84） in a dosage of 1, 2 and 4 μg/kg respectively. Blood samples were obtained before and after administration within 24 hours. The rhPTH concentrations in sennn were determined by enzyme linked immunosorbent assay （ELISA）. The pharmacokinetic parameters determined with use of standard noncompartmental analysis were the maximum serum concentration （ Cmax ）, the time to attain that concentration （ tmax ）, and the area under the serum concentration-time curve up to 24 hours（ AUC0-24 ） and up to infinity （AUC0-∞）. Dose proportionality of pharmacokinetic parameters （AUC, Cmax of every volunteer of each dosage and A UC was computed from log transformed data） and was examined by mean of analysis of variance （ANOVA） using SPSS software package. In the study, subjects＇ symptoms, objective signs, and vital signs, including blood pressure, heart rate, respiratory rate and body temperature, were checked and 12-lead electrocardiography was recorded before and after drug administration within 24 hours. Routine laboratory tests, including hematology, blood biochemistry, serum electrolyte, and urinalysis, were performed before and after drug administration within at 24 hours.[第一段]     

Regulators of Cardiac Cell Growth, Differentiation, and Apoptosis

The pathogenesis of heart hypertrophy and failure have been the focus of intense clinical and basic science investigation, yet the signal transduction pathways and molecular process that underlie the compensatory growth process that ultimately leads to heart failure remain enigmatic. Since ventricular myocytes have exited the cell cycle, growth of the myocardium in response to hemodynamic load occurs by cellular hypertrophy and not by hyperplasia. In this article, we document the potential involvement of tumor suppressor pocket proteins and cell cycle regulators that may impinge on the growth, differentiation, and apoptosis of cardiac muscle. This revised version was published online in August 2006 with corrections to the Cover Date.     

Benign intracranial hypertension and recombinant growth hormone therapy in Australia and New Zealand

Benign intracranial hypertension (BIH) is reported in three children from Australia and one from New Zealand, who were being treated with recombinant human growth hormone (rhGH). Three males and one female, aged between 10.5 and 14.2 y, developed intracranial hypertension within 2 weeks to 3 months of starting treatment. A national database, OZGROW, has been prospectively collecting data on all 3332 children treated with rhGH in Australia and New Zealand from January 1986 to 1996. The incidence of BIH in children treated with growth hormone (GH) is small, 1.2 per 1000 cases overall, but appears to be greater with biochemical GHD (<10IUml ^-1), i.e. 6.5/1000 (3 in 465 cases), relative risk 18.4, 95% confidence interval 1.9-176.1, than in all other children on the database. The incidence in patients with Turner's syndrome was 2.3/1000 (1 in 428 cases). No cases in patients with partial GHD (10–20 IUml ^-1) or chronic renal failure were identified. Possible causative mechanisms are discussed. The authors'practice is now to start GH replacement at less than the usual recommended dose of 14IUm-² week^-1 in those children considered to be at high risk of developing BIH. Ophthalmological evaluation is recommended for children before and during the first few months following commencement of rhGH therapy and is mandatory in the event of peripheral or facial oedema, persistent headaches, vomiting or visual symptoms. The absence of papilledema does not exclude the diagnosis.     

重组促红细胞生成素对腹膜透析患者心功能的影响

目的　了解基因重组人促红细胞生成素 (r HuEPO)对持续性非卧床腹膜透析 (CAPD)患者心室结构及功能的影响。方法　 32例CAPD伴高血压患者 ,其中 1 6例应用降压药物控制血压 ,同时应用r HuEPO纠正贫血 ,1 6例单用降压药物 ,随访 6个月。结果　两组治疗前后差值比较 ,治疗组的LVPWT、E/A与对照组比较有显著差异(P <0 .0 5 )。结论　长期纠正贫血和控制血压有利于透析患者左室结构及功能异常的逆转     

人IL-10基因重组复制缺陷型腺病毒DNA的构建

目的 构建人IL-10基因的重组腺病毒并进行体外表达和检测。方法从质粒pcDNA3IL-10的CMV启动子下游完整切下IL-10 cDNA的片段,将其定向插入Gateway载体,在克隆酶的作用下将阳性克隆质粒转入目的腺病毒载体。PacI酶线性化IL-10腺病毒DNA后阳离子质脂体转染293A细胞,获得人IL-10的复制缺陷型重组腺病毒。体外感染人胰腺癌细胞株Bxpc-3和大鼠胰腺细胞株AR-42J;Western blot检测人IL-10的表达。结果 成功地构建人IL-10重组腺病毒,病毒滴度达2×109PFU/mL。体外感染的细胞株均检测到IL-10的表达。结论 构建复制缺陷型重组腺病毒能够介导IL-10的基因表达,为细胞因子的抗炎冶疗奠定实验基础。     

腺病毒介导钙离子ATP酶基因在骨髓间充质干细胞的表达

目的：以腺病毒（Ad—CMV）介导钙离子ATP酶基因（sarcoplasmic reticulum Ca^2＋ ATPase，SERCa2a）转染骨髓间充质干细胞（Bone marrow msenchymal stem cells，BMSCs），测定表达产物，为心力衰竭的细胞移植和基因治疗提供实验基础。方法：体外培养大鼠骨髓间充质干细胞，用已构建好的Ad5-CMVSERCa2a真核载体进行转染。通过流式细胞仪检测转染效率。逆转录-聚合酶链反应（RT-PCR）、Westem杂交等方法检测SERCa2amRNA和蛋白质的表达。结果：Ad5-CMV—SERCa2a对BMC的转染率为80％左右，没有明显的细胞毒性反应。RT-PCR检测到SERCa2a，Western杂交确定SERCa2a蛋白表达。结论：腺病毒载体可有效地介导SERCa2a基因转染BMC，并表达SERCa2a蛋白，为基因治疗心力衰竭的研究奠定了基础。     

携带人胰岛素样生长因子-1重组腺病毒的构建和鉴定

[目的] 采用 Cre-LoxP同源重组系统构建并鉴定携带人胰岛素样生长因子-1( human insulin-like growth factor-1, hIGF-1)目的基因的复制缺陷重组腺病毒,为椎间盘退变的 hIGF-1基因治疗奠定基础.[方法] PCR合成 hIGF-1基因,用 pMD18-T载体克隆 hIGF-1目的基因,酶切、测序并进行 NCBI BLAST相似性在线分析.将 pMD18-T-hIGF-1和 pDNR-1r质粒分别行 EcoRⅠ和 BamHⅠ双酶切, DNA 连接酶连接双酶切产物,转化感受态大肠杆菌 DH5α,合成中间载体 pDNR-hIGF-1,酶切鉴定.Cre-loxP系统介导 pDNR-hIGF-1和 pInt.AV1.SpaT同源重组形成 pInt.AV1.SpaT-hIGF-1重组表达质粒,行 EcoRⅠ酶切鉴定.复苏 293细胞,将 pInt.AV1.SpaT-hIGF-1和 pInt.B.B质粒在 293细胞内进行同源重组成含 hIGF-1基因的复制缺陷重组腺病毒.倒置显微镜观察 293细胞病变样效应( cytopathogenic effect, CPE);透射电镜观察 293细胞中的病毒颗粒;提取细胞裂解液中病毒 DNA, PCR鉴定 hIGF-1目的基因的存在; Western blot 检测 hIGF-1蛋白表达.用半数组织培养感染量 (tissue culture infectious dose 50,TCID50)方法测定重组腺病毒的滴度.[结果]克隆的 hIGF-1目的基因经 NCBI BLAST相似性在线分析证实与 Homo sapiens IGF-1 (GI:19923111)完全相同.pInt.AV1.SpaT-hIGF-1酶切鉴定,出现 5.4 kb和 1 kb两条片段,与理论值完全相符.转染 293细胞 12～ 14 d后,大部分细胞出现肿胀,脱落等细胞病变样效应.PCR鉴定细胞裂解液中含有 hIGF-1目的基因.Western blot 证实重组腺病毒表达 hIGF-1蛋白.第 2代腺病毒的滴度为 80× 106 PFU/mL.[结论]成功构建出含有 hIGF-1目的基因的复制缺陷重组腺病毒.     

抗戊型肝炎病毒重组蛋白单克隆抗体的制备和初步应用

目的:制备抗-戊型肝炎病毒的单克隆抗体,并将其用于分析戊型肝炎病毒不同毒株结构蛋白的抗原表位。方法:采用来自墨西哥株(Mexicanstrain)的戊型肝炎病毒重组蛋白(p166Mex)免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经酶联免疫吸附试验(ELISA)法筛选阳性克隆,并将获得的单克隆抗体与戊型肝炎病毒缅甸株(Burmastrain)和美国株(USAstrain)的重组蛋白(p166Bur、p166US)进行交叉反应测定。结果:最终获得4株能稳定分泌抗-p166Mex的杂交瘤细胞株,即D8G10、E5E12、D4A3、B7E6。其中D8G10,E5E12和B7E6细胞株的培养上清液,还能分别与p166Bur和p166US重组蛋白发生阳性反应。结论:利用已获得的抗-p166Mex单克隆抗体,初步确定3种不同的戊型肝炎病毒重组蛋白(p166Bur、p166US、p166Mex)含有一种共同的抗原表位。     

生长激素抗肝纤维化的作用及其机制

目的　探讨重组人生长激素 (rhGH)对肝硬化大鼠在肝功能、门静脉高压等方面的治疗作用及其可能的机制。方法　雄性SD大鼠建立肝硬化模型 ,随机分组 ,分别给予NS、rhGH (333ng/KgBW ,ip ,qd× 7d)处理。 结果　rhGH处理前后肝硬化肝组织GHR的配体结合容量 [fmol/mg ,(31± 4 )vs .(4 0± 7) ]及其mRNA量 (iOD ,pixel)均显著升高 (2 3± 3)vs.(4 2± 8) ;P <0 0 5 ,肝硬化大鼠血清白蛋白 (g/L )显著升高 (2 9± 4 )vs .(37± 7) ;P <0 0 5、ALT活性 (U/L)显著降低 (89± 15 ,6 9± 7;P <0 0 5 ) ,肝硬化肝组织丙二醛含量 (nmol/mg)显著降低 (18 7± 3 2 ,12 0± 2 2 ;P <0 0 5 )、超氧化物歧化酶活性 (U/mg)显著升高 (82 4± 10 8,10 2 9± 76 ;P <0 0 5 )、胶原纤维相对含量 (% )显著降低 (2 2 30± 3 86vs.14 70± 2 0 7;P <0 0 5 ) ,肝硬化大鼠的门静脉压 (cmH2 O)亦显著降低 (14 4± 2 0vs.9 3± 1 5 ;P <0 0 5 )。结论　药理剂量的rhGH能够促进肝硬化大鼠的白蛋白合成、改善肝细胞功能 ,有助于抑制肝组织纤维化、缓解门静脉压力。     

Allergen microarray: comparison of microarray using recombinant allergens with conventional diagnostic methods to detect allergen-specific serum immunoglobulin E

BACKGROUND: The availability of recombinant allergens and recent advances in biochip technology led to the development of a novel test system for the detection of allergen-specific IgE. OBJECTIVE: To test the performance of this allergen microarray in a serological analytical study. METHODS: Standard allergens contained in grass pollen (Phl p 1, Phl p 2, Phl p 5 and Phl p 6) and tree pollen (Bet v 1 and Bet v 2) were used as a model system. The detection of allergen-specific serum IgE using microarrays was compared with standard test systems: CAP/RAST and an in-house ELISA. In order to test the analytical sensitivity of the assays, geometric dilutions of a serum pool containing high levels of pollen-specific IgE from allergic individuals were tested in each system. To assess the analytical specificity, the sera of 51 patients with presumptive allergic symptoms were collected before diagnosis. Thereafter, the results for grass/tree-pollen-specific IgE were compared. RESULTS: The microarray has a good dynamic range similar to the CAP/RAST system. Microarray and ELISA showed comparable analytical sensitivity exceeding the CAP/RAST system. With respect to the analytical specificity, no significant cross-reactivity of the allergens was observed. For two of the allergens tested, weak positive signals were detected in the microarray test system, whereas they were not detectable by CAP/RAST. CONCLUSION: A good correlation of presently used methods to detect serum IgE and the novel microarray test system was observed. As a next step, a careful validation of this method for a multitude of allergens and a thorough clinical evaluation has to be provided. Microarray testing of allergen-specific IgE can be presumed to be the method of choice for a prospective component-resolved diagnosis of Type I allergy, and the basis for the design and monitoring of a patient-tailored specific immunotherapy in the future.