Primary structure of seiwhale pituitary somatotropin

Seiwhale somatotropin has been isolated from seiwhale pituitaries. It was cleaved by cyanogen bromide, trypsin and chymotrypsin. The peptide fragments were separated and purified by gel filtration on Sephadexes, ion exchange chromatography, high voltage electrophoresis and paper chromatography. Amino acid sequences of the isolated peptides were studied by the dansyl-Edman procedure. The data obtained suggested a primary structure of seiwhale somatotropin consisting of 190 amino acid residues and showed a high degree of homology with somatotropins of many other species.     

Contact area of bovine somatotropin dimer: involvement of tyrosine 142

The presence of tyrosine residues in the contact area between protomers of bovine somatotropin dimers (Fernandez & Delfino, Biochem. J. 209 , 107-115, 1983) was investigated taking advantage of the impaired self-associating ability of molecules iodinated at such residues. Reaction of bovine somatotropin dissolved in 8 m urea with the NaI-Chloramine T couple (2.1 × 10^?4m ) rendered a preparation with 3.1 iodine atoms per molecule which, by stepwise elimination of the denaturant and gel filtration through Sephadex G-100, originated two distinguishable populations: one able (iododerivatives I), the other unable (iododerivatives II) to self-associate. After frontal analysis, iododerivatives II were found to be unable to interact even with native molecules. Identification of the reacting tyrosine residues indicated that iodination of tyrosine 142 was responsible for the loss of the ability to form dimers in iododerivatives II. Iodohormones retained the ability to bind to somatogenic mouse hepatocyte receptors - the relative potency for iododerivatives I and II being 0.60 (0.34–1.03) and 0.71 (0.41–1.22) respectively.     

Measurement of biological activity of somatotropin in hypophysectomized rats

目的：建立测定生长激素（ＧＨ）在体生物活性的方法．方法：以去垂体大鼠体重增长（ＢＷＧ）和胫骨骺软骨板宽度（ＴＥＷ）为指标，观察动物性别、给药途径、次数和周期不同对效应的影响；同时进行４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法，测定ＧＨ的效价（平行线３×３设计）．结果：♀和♂ｓｃ和ｉｍ给药以及每日给药１次和２次的ＢＷＧ和ＴＥＷ差异无显著意义．给药６ｄ比给药４ｄ引起较大的ＢＷＧ和ＴＥＷ（Ｐ＜００５）．４ｄＢＷＧ法和６ｄＢＷＧ法在００２０－０５００ＩＵ·ｄ－１有较好的λ值（００６６０和０１７４７）和ｒ值（０９０００和０９２３７）；４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法测得ｒｈＧＨ的效价为４６１３２，３９８２９和４８０２３ＩＵ／ａｍｐ．６ｄＢＷＧ法有较小的λ值和较低的ＡＲＦＬ值．结论：可在同一组去垂体大鼠体内同时用４ｄＢＷＧ，６ｄＢＷＧ和６ｄＴＥＷ法测ＧＨ活性，以６ｄＢＷＧ法较好．     

Carboxylate groups in bovine somatotropin involved in growth promoting activity*

Modification of approximately one fifth of the carboxylate groups in bovine somatotropin with a water soluble carbodiimide caused loss of growth promoting potency pointing to the existence of residues related to the hormonal activity among those belonging to a fast reacting set. A sigmoidal curve was obtained whether the inactivation process was referred to reaction time or degree of modification. Isoelectrofocusing of derivatives released the native hormone from responsibility for the biological potency exerted by preparations with 1.5–2.6 modified carboxylate groups. Examination of the individual reaction kinetics of the 11 fast reacting residues, in turn, excluded the possibility of the sigmoidal character of the inactivation curve being caused by a nonexponential disappearance of essential residues, as a possible consequence of the chemical modification of others. According to synthetic models, the experimental curve may be the consequence of the effect of cumulative modification of 2 or 3 out of a set of 3 to 8 relevant residues.     

Human β-endorphin

Three analogs of human β-endorphin (β_h-EP) were synthesized by the solidphase method: [Gln⁸, Trp²⁷]-β_h-EP (I), [Gln⁸,Arg⁹,Trp²⁷]-β_h-EP (II), and [Gln⁹,Arg¹¹,Trp²⁷]-β-EP (III). Radioreceptor binding assay with use of tritiated β_h-EP as primary ligand gave relative potencies as follows: β_h-EP, 100; I, 778; II, 467; III, 449. Relative potencies in an analgesic assay were: β-EP, 100; I, 114; II, 165; III, 83. The 8–11 segment of β_h-EP can tolerate a net increase in charge of + 2 without diminishing analgesic potency. The substitution of Glu⁸ may be one of the more dependable means of designing β-endorphin antagonists.     

Serum responsiveness to recombinant bovine somatotropin in buffalo: a three‐month lactation study using an acid‐stripping ELISA for screening

The misuse of recombinant bovine somatotropin (rbST) to increase milk yield involves buffalo not just cows. Screening methods to identify rbST‐treated cattle have already been proposed. However, there have been no studies on prolonged periods with a high number of animals. In this study, we developed a new enzyme‐linked immunosorbent assay (ELISA) to measure the serum responsiveness towards rbST, based on an acid‐stripping procedure and relatively simple integral calculation dilution curves. We also applied the analysis to 640 serum and 96 milk samples collected from 16 buffalo treated with rbST and 16 controls, over a period of approximately three months. Its suitability as a screening method, in compliance with EU law, was also assessed. A bi‐factorial approach was also evaluated, including the measurement of insulin‐like growth factor 1 concentration in serum. Results showed that our ELISA could be used on its own for screening purposes, without the need to assess other biomarkers. Copyright © 2016 John Wiley & Sons, Ltd.     

Human thrombin receptor-activating peptide-induced proliferation of cultured vascular smooth muscle cells exhibits species specificity

Thrombin receptor stimulation in vitro signals many cellular events that are associated with the response to vascular injury in vivo. Indeed, we have previously shown that human α-thrombin and the 14-amino acid human thrombin receptor-activating peptide (huTRAP-14) stimulate proliferation of cultured rat aortic smooth muscle cells (SMC). In the present studies, the mitogenic response of rabbit vascular SMC to thrombin and huTRAP-14 was assessed using [³H]thymidine incorporation and cell number. Results demonstrated that thrombin stimulated mitogenesis of rabbit vascular SMC in culture and that the thrombin response was dependent on proteolytic activity. However, huTRAP-14 was not mitogenic for rabbit vascular SMC. Thus, there are species differences in huTRAP-14 responsiveness. As rat and rabbit models continue to be used extensively to evaluate mechanisms and potential therapies for human restenosis, it is important to identify any species differences in the mechanism whereby thrombin exerts its biological effects. © 1995 Wiley-Liss, Inc.     

第二批人凝血酶国家标准品协作标定

目的协作标定第二批人凝血酶国家标准品。方法用世界卫生组织首批α 凝血酶国际标准品 (批号 :89/ 5 88)为标准 ,采用纤维蛋白原凝固法标定。并将标准品分别于 2 5℃、37℃、4 5℃保存 6个月 ,进行稳定性考查。结果第二批人凝血酶国家标准品的效价为 12 7IU/支 ,稳定性良好 ,其他项目符合国家标准品要求。结论已成功研制第二批人凝酶国家标准品 ,可以取代首批人凝血酶国家标准品。     

用HPLC法测定人凝血酶制品中甘氨酸的含量

目的 建立一种测定人凝血酶制品中甘氨酸含量的HPLC方法。方法 以2、4-二硝基氟苯（DNFB）为柱前衍生剂,采用ODS-C₁₈色谱柱,流动相为50%乙腈溶液-0.05 mol/L醋酸钠缓冲液（35︰65）,流速1.0 ml/min,检测波长360 nm。结果 甘氨酸的线性范围为0.006~0.030 mg/ml（r=0.999 6）,平均回收率为100.4%,RSD为0.44%（n=9）。结论 该方法简便、准确、专属性好,可用于人凝血酶制品中甘氨酸的含量测定。     

Human retinoblastomas have binding sites for the COOH-terminal segment of human β-endorphin

Two human retinoblastoma cell lines (Y79 and McA) were evaluated for the presence of binding sites for human beta-endorphin (β_h-EP). Using tritiated β_h-EP (³H-β_h-EP) and synthetic β-EP analogues, it was possible to demonstrate binding sites for ³H-β_h-EP with an ED₅₀ of 3.5 nM in Y79 cells and 8 nM in McA cells respectively. The non-opioid segment [β_h-EP-(6–31)] retained about 20% relative potency in Y79 and 40% in McA in displacing the tritiated hormone when compared with β_h-EP. Camel β-EP had a relative potency of less than 1% and β_h-EP-(1–27) was inactive in both cells in doses as high as 4μm. Taken together with previous reports on similar binding sites in human neuroblastoma and glioblastoma cell lines, it appears that cell lines of neural origin have binding sites for the COOH-terminal of human β-EP.     

Assessment of alpha1-adrenoceptor antagonists in benign prostatic hyperplasia based on the receptor occupancy theory

AIMS: To assess the mechanistic relationship between doxazosin (alpha(1)-receptor antagonist) and receptor occupancy and a measure of pharmacological effect (Q(max), the maximum urinary flow rate) and to compare the mean receptor occupancy ratio at clinical doses of doxazosin, tamsulosin, terazosin and prazosin in benign prostatic hyperplasia (BPH). METHODS: A ternary complex model, which described the mechanism of alpha(1)-receptor antagonists, was fitted to the pharmacological effects and receptor occupancy ratio data for doxazosin (standard tablet). In addition, mean receptor occupancy was calculated for other alpha(1)-receptor antagonists and the optimal receptor occupancy was evaluated. The clinical pharmacological effects of the controlled release formulation of doxazosin (doxazosin GITS) were estimated based on the receptor occupancy. RESULTS: The mechanistic based model was able to describe the pharmacological effects of doxazosin. Regardless of the plasma concentrations or clinical dose of each drug, the results suggest that receptor occupancy is useful to assess quantitatively and compare the pharmacological effects of drugs with similar mechanisms of action. The clinical dosage for doxazosin GITS was estimated to be at least 8 mg and the stable pharmacological effect is expected based on the estimated receptor occupancy. CONCLUSIONS: A model for Q(max) improvement in BPH based on the receptor occupancy theory was able to describe the clinical effects of the alpha(1)-receptor antagonists. Receptor occupancy is a useful index for predicting the clinical effects of alpha(1)-receptor antagonists.     

Primary structure of fox pituitary growth hormone

Tryptic digests of fox growth hormone (fGH) were separated by reverse phase high performance liquid chromatography (HPLC) and by paper electrophoresis. From the amino acid composition of these tryptic peptides and from their alignment with the expected tryptic peptides from bovine growth hormone (bGH), the primary structure of fGH is proposed. There are only 17 amino acid residues which are different in these two growth hormone molecules.     

Primary structure of elephant pituitary prolactin

Tryptic digests of elephant pituitary prolactin (elePRL) were separated by reverse phase high performance liquid chromatography (HPLC) and paper electrophoresis. From the amino acid composition, the amino acid sequencing of selected peptides, and from their alignment with expected tryptic peptides from ovine prolactin (oPRL), the primary sequence of elePRL is proposed.     

Human embryonic stem cells and lung regeneration

Human embryonic stem cells are pluripotent cells derived from the inner cell mass of preimplantation stage embryos. Their unique potential to give rise to all differentiated cell types has generated great interest in stem cell research and the potential that it may have in developmental biology, medicine and pharmacology. The main focus of stem cell research has been on cell therapy for pathological conditions with no current methods of treatment, such as neurodegenerative diseases, cardiac pathology, retinal dysfunction and lung and liver disease. The overall aim is to develop methods of application either of pure cell populations or of whole tissue parts to the diseased organ under investigation. In the field of pulmonary research, studies using human embryonic stem cells have succeeded in generating enriched cultures of type II pneumocytes in vitro. On account of their potential of indefinite proliferation in vitro, embryonic stem cells could be a source of an unlimited supply of cells available for transplantation and for use in gene therapy. Uncovering the ability to generate such cell types will expand our understanding of biological processes to such a degree that disease understanding and management could change dramatically.     

论人体生命的三重品格

本文讨论的是人体生命观的问题,从人的本质来看,人体生命与其它生命体有着原则的区别。人体生命是神圣品格、质量品格和价值品格的辩证统一。人体生命三重品格以价值品格为核心,密不可分地联系在一起。片面强调人体生命的质量品格或神圣品格,必然使人们在理论和实践上陷入二难境地。     

β-Endorphin: synthesis and properties of analogs modified in positions 8 and 31

Four analogs of human β-endorphin (β_h-EP) were synthesized by the solid-phase method: [Gln^8.31]-β_h-EP(I), [Arg⁸, Gln³¹] -β_h-EP(II), [Ala⁸, Gln³¹] -β_h-EP (III), and [Val⁸, Gln³¹]-β_h-EP(IV). Radioreceptor binding assay with use of tritiated β_h-EP as primary ligand gave relative potencies as follows: β_h-EP, 100; I, 200; II, 150; III, 150; IV, 120. Relative potencies in an analgesic assay were: β_h-EP, 100; 1,236; II, 254;III, 116; IV, 121. The side-chain of Glu-8 in β_h-EP can be replaced by a variety of structures without diminishing biological activity.     

THE THROMBIN RECEPTOR

1. The thrombin receptor has now been cloned and found to be a member of the G-protein-coupled seven-transmembrane domain receptor family. 2. The receptor has been detected directly in platelets, endothelial cells and smooth muscle cells and studies using receptor-derived peptides have demonstrated that this receptor may be the one responsible for many of the actions of thrombin in platelets, endothelial cells, smooth muscle cells, fibroblasts, mesangial cells and neural cells. 3. The receptor appears to be activated by the novel mechanism of cleavage by thrombin to yield a new N-terminus which then interacts with the receptor as a tethered ligand to initiate cell activation.     

ß-endorphin: synthesis and properties of analogs modified at position 6

Two analogs of ß-endorphin (ß-EP) were synthesized by the solid-phase method: [D-Ser⁶]-ß_c-EP (I) and [Pro⁶]-ß_c-EP (II)(ß_c-EP = camel ß-EP). Radioreceptor binding assay with use of tritiated human ß-EP (ß_h-EP) as primary ligand gave relative potencies as follows: ß_c-EP, 100; I, 32; II, 28. Relative potencies in an analgesia assay were: ß_c-EP, 100; I, 115; II, 31.     

我院人用狂犬病疫苗及狂犬病人免疫球蛋白使用情况分析

目的：总结分析2015-2017年我院人用狂犬病疫苗及狂犬病人免疫球蛋白（HRIG）的使用情况,为科学、合理地制定采购计划及储备管理提供依据。方法：通过医院信息管理系统调取2015-2017年我院人用狂犬病疫苗及HRIG的相关使用数据,计算其月占比、季节指数等数据,对我院2015-2017年人用狂犬病疫苗及HRIG的使用情况进行分析。结果：2015-2017年我院人用狂犬病疫苗共使用484166支,分别为145140支、154818支、184208支,使用总量逐年增加。其中5-10月使用量最大,季节指数均超过100%;12月至来年3月接种量使用量相对较小,季节指数均不超过80%。3年间HRIG的使用总量也逐年明显增加,季节指数的变化与人用狂犬病疫苗相似。结论：2015-2017年我院人用狂犬病疫苗及HRIG的使用量基本呈现夏秋季多、冬春季少的规律,具有典型的季节性。根据时间序列方程式计算得到的2018年人用狂犬病疫苗预测使用数量与实际使用数量并不相符,误差率较大,易受外界因素影响。因此,还是要根据季节及实际使用情况的变化,动态地进行人用狂犬病疫苗及HRIG的库存管理。人用狂犬病疫苗在使用中发现的不良反应多为局部反应,但我院近年来也偶发非常严重的不良反应,因此要加强观察,提高重视。     

UHPLC-LTQ-Orbitrap-MS分析人血白蛋白及人免疫球蛋白中非目标蛋白成分

目的 采用超高效液相色谱-串联线性离子阱-轨道阱组合高分辨质谱技术对人血白蛋白及人免疫球蛋白中非目标蛋白成分进行定性分析。方法 采用ACQUITY UPLC peptide BEH C₁₈(300Å,1.7 μm,2.1 mm×100 mm)色谱柱,以0.1%甲酸水溶液-0.1%甲酸乙腈为流动相进行梯度洗脱,质谱采用Full MS/dd-MS²(TopN)扫描模式。结果 从人血白蛋白及人免疫球蛋白中共鉴定出非目标蛋白52个,其中人血白蛋白样本中鉴定非目标蛋白25个,人免疫球蛋白样本中鉴定非目标蛋白27个。结论 建立的定性方法可快速、准确、系统地识别出人血白蛋白及人免疫球蛋白中多种蛋白,为该类制剂质量控制及进一步的临床应用提供参考。