蛋白质药品的真空冷冻干燥技术及研究进展

目的介绍真空冷冻干燥技术制备蛋白质药品的方法。方法查阅国内外近年来有关文献，综述了真空冷冻干燥原理、蛋白质药品的冻干工艺及药品贮存期间影响其活性的各个因素。结果真空冷冻干燥过程中可引起蛋白质药品不同程度的变性，加入保护剂可增加其稳定性。结论冷冻干燥时，不同的蛋白质药品需加入不同的保护剂。     

蛋白质内含子研究进展

介绍了蛋白质内含子的结构、功能和其介导的蛋白质剪接机理及应用研究进展。蛋白质内含子目前已广泛应用于蛋白质纯化、蛋白质连接、以及环肽和毒素蛋白的制备,且对蛋白质结构和功能关系的研究具有重要价值。     

基于蛋白质序列预测蛋白质-蛋白质相互作用位点研究进展

生物分子之间的相互作用是生命过程的分子基础,其中蛋白质分子之间的相互作用占有极其重要的地位。因此研究蛋白质相互作用对于理解生命的分子机理、探讨致病微生物的致病机理,以及研究新药提高人们的健康水平具有重要的作用。文章对蛋白质-蛋白质相互作用数据库和蛋白质-蛋白质相互作用位点的序列特征和蛋白质-蛋白质结合位点预测最先进的方法进行介绍。蛋白质-蛋白质相互作用位点的残基在调节物理结合过程中具有重要的意义,例如这些残基参与酶抑制剂相互作用的抑制效应,通过抗体-抗原相互作用的最初免疫反应,和细胞中信号蛋白的调节等。当前,各种各样的方法被用于预测蛋白质-蛋白质相互作用位点的预测中,并通过整体的构建蛋白质-蛋白质作用网络,从残基水平去理解蛋白质结合现象。     

人纤维结合蛋白制备方法的建立及人纤维结合蛋白稳定性初探

目的  建立人纤维结合蛋白（fibronectin,Fn）制备方法,研究不同保护剂对Fn稳定性的影响。方法  将经磷酸盐缓冲液抽提溶解的血浆冷沉淀用聚乙二醇沉淀,获得的上清液经离子交换层析制得纯化的Fn。将加入复合保护剂的Fn溶液分别于-20、4和37 ℃存放0、5和14 d,观察保护剂抑制Fn降解的效果。结果  采用建立的方法制备的Fn的纯度和平均回收率均>80%。Fn稳定性研究显示,Fn于4℃存放会发生降解,加入枸橼酸钠+甘氨酸或枸橼酸钠+咪唑复合保护剂能有效抑制Fn降解。结论  采用建立的方法制备Fn是可行的。枸橼酸钠+甘氨酸或枸橼酸钠+咪唑复合保护剂对Fn具有保护作用。     

蛋白质组学在药物研究中的应用

蛋白质组是指基因组表达的所有相应的蛋白质，是指细胞或组织或机体全部蛋白质的存在及其活动方式。蛋白质组具有多样性和可变性，同一机体的不同细胞中，蛋白质的种类和数量是各不相同的，即使是同一种细胞。在不同时期、不同生理条件下，其蛋白质组都是在不断变化之中，在病理过程或药物作用下，细胞蛋白质的组成及其变化，与正常生理过程也是不同的。蛋白质组学是从整体的蛋白质水平上，从生命本质的层次上，研究和发现生命活动的规律和重要生理、病理现象的本质。蛋白质的研究技术主要应用于两个方面，一个是特殊细胞、组织或有机体的系统鉴定和所有蛋白的定量，这是系统生物学的核心，可以提供一个完全量化的含翻译后修饰变体的蛋白质组，更重要的是可以在相关样品中寻找差异，即在生理状态发生变化而产生的蛋白质图谱中的差异。另一个方面主要是蛋白质功能和蛋白质相互作用的研究，包括如蛋白质序列、结构、相互作用和生化活性的分析等多种试验方法。药物蛋白质组学的重要研究内容在临床前包括新药和靶的发现、药物作用模式、毒理学研究，在临床研究方面包括疾病特异性蛋白作为有效患者选择的依据和临床试验的标志。应用类似于药物遗传学的方法，按照蛋白质谱来分类患者，并预测药物作用疗效。     

高蛋白营养支持对重症颅脑外伤低能量摄入时蛋白质代谢的影响

目的探讨低能量摄入时给予高蛋白质营养支持对重型颅脑损伤患者蛋白质代谢的影响。方法将25例重型颅脑损伤患者随机分为蛋白质摄入为0.8 g/kg体重的常规蛋白质组(n=12)和摄入2.0 g/kg体重的高蛋白质组(n=13),在低能量摄入时测定两组患者在营养支持开始前、营养支持7 d、14 d时血中转铁蛋白、前白蛋白和氨基酸浓度以及氮平衡。结果常规蛋白质组在营养支持7 d和14 d时,血中转铁蛋白和前白蛋白含量较低,氮平衡回升较慢,高蛋白质组血中转铁蛋白和前白蛋白含量较高,氮平衡回升较快,在营养支持7 d时,两组多数血清氨基酸含量较营养支持前降低,在营养支持14 d时,高蛋白质组已有较高的氨基酸含量。结论脑外伤患者在低能量摄入时给予充足的蛋白质有利于促进蛋白质的合成代谢。     

包封和释放过程中稳定蛋白质结构的研究进展

药用蛋白质从生物可降解聚合物中释放的过程始终影响蛋白质的三维结构、化学性质和完整性。在使用最常用的水／油／水（W／O／W）双重乳化技术包封蛋白质时，要想保持其稳定性就需要提高蛋白质的热力学稳定性或者通过使用各种添加剂使蛋白质避免接触失稳剂（如W／0界面）。然而，当人们使用W／O／W技术时，蛋白质的稳定性仍存在问题。因此，探索其他解决这种问题的方法就受到了关注。这些替代方法，如固／油／油（S／O／O）和固／油／水（S／O／W）技术是基于将干燥的蛋白质粉末悬浮于无水有机溶剂中的一种技术。很明显，蛋白质结构在有机相中的稳定性是由于蛋白质在此状态下是“僵硬的”，因而蛋白质结构的干燥是禁阻的。本文主要介绍在应用各种不同的包封技术时如何保持蛋白质结构和性能的稳定性。     

蛋白质、多肽类药物制剂的研究现状

目的：综述蛋白质、多肽类药物制剂的研究现状。方法：依据国内外文献进行综述,包括蛋白质、多肽类药物性能特点、已开发剂型及其制备方法,临床应用效果和发展前景内容。结果：蛋白质、多肽类药物制剂的研究是目前蛋白质、多肽类药物发展的迫切要求,但目前研究工作较少,理论基础研究则更少。结论：蛋白质、多肽类药物将成为21世纪的重要药品,我们应该加强蛋白质、多肽类药物制剂的的研究,使其更适合于临床应用的要求。     

食物蛋白质资源-猪血浆蛋白质的营养学研究

本文用化学分析与动物实验的方法,对昆明植物研究所和昆明轻工研究所从猪血中分离提取猪的血浆蛋白质,猪血球蛋白质,猪脱色血球蛋白质,进行了营养学方面的研究。结果表明,该两单位研制生产的猪血浆蛋白质营养价值较高,可代替全鸡蛋广泛运用于食品工业的生产中。     

蛋白质小知识

1克蛋白质产生4千卡的能量。 构成人体蛋白质的有20种氨基酸。其中，9种为必需氨基酸。必需氨基酸指人体不能合成，必须从食物中直接获得的氨基酸。这9种氨基酸是：异亮氨酸、亮氨酸、赖氨酸、蛋氨酸、苯丙氨酸。     

The debate on DDT

The paper reviews the early toxicologic and pharmacologic studies carried out by the author and his associates from 1943 to 1947, which were largely responsible for launching DDT as an agent for the control of typhus, malaria, yellow fever, and related vector-borne diseases. After reviewing recent studies conducted at the University of Miami, which dealt with organochlorine pesticides in human tissues, the tumorigenicity of aldrin, dieldrin and endrin (rat), six-generation mouse and three-generation dog reproduction studies, synergism of DDT and aldrin (dog), and the fate of DDT and aldrin during a period of severe starvation (rat), it is pointed out that it is primarily the overuse and misuse of DDT in pest control that have caused the pollution in our ecology. It is emphasized that the requirements for pest control differ the world over and that it must therefore be left to the national regulatory agencies to legislate the safe use of DDT and related pesticides. It is recommended that future human and animal studies with DDT and its derivatives give consideration to: (a) the balance and metabolism of the various hormones, (b) reproduction (estrus, libido, mammary development, milk production, (c) hepatic microsomal enzyme activities, (d) cancer prevention and cancer production, (e) excessive body weight changes induced by disease, unbalanced diet or starvation, and (f) the effects of DDT and its derivatives when absorbed in combination with other related and even unrelated compounds.Presented at the joint meeting of the Scandinavian and German Pharmacological Societies, Copenhagen, Denmark, July 20–23, 1971.     

肠道菌群与健康、疾病和药物作用的影响

肠道菌群作为人体内一个复杂的微生态系统,在维持人体微生态的稳态中,肠道菌群在维持宿主生理功能具有上非常重要的作用,也对许多代谢性疾病、免疫性疾病以及肿瘤都有着密切的关系,且对于药物治疗合理安全有效具有重要意义。本文从正视存在人体的细菌的有益性和有害性、肠道菌群与健康和寿命、肠道菌群与疾病以及药物作用的影响等4方面分析和讨论。肠道菌群与不同类型药物的关系已经成为近些年的热点研究领域,本文分别讨论免疫治疗、化学药物、抗生素和中药的相关问题,希望为认识药物治疗过程、科学合理用药、认识药物作用机制、新药研究开发等研究有所参考。     

海星皂苷生物活性与制备研究进展

海洋是全球药物研发的重要宝库，提高海洋生物资源深度开发和高值化利用能力，是我国海洋强国战略的重要组成部分，也是促进海洋经济可持续发展及实施“蓝色药库”的关键途径之一。海星属典型的棘皮动物，进化地位和生物学特征独特，是国际公认的药用/保健用海洋生物。海星中含有皂苷、多糖、多肽、氨基酸、胶原蛋白、甾醇及生物碱等多种营养成分和活性物质，其中海星皂苷在抗肿瘤、抗炎、抗衰老及降血脂等方面展现出良好的生物活性，在食品和药物研发领域具有巨大的发展潜力和广阔的应用前景。本文系统检索了近30年海星皂苷的研发现况，并且对近15年来海星皂苷的生物活性、提取分离及相关专利等方面取得的研究进展进行梳理，进而为其在营养保健和药物研发中的应用提供相关理论支持。     

Molecular handling of cadmium in transporting epithelia

Cadmium (Cd) is an industrial and environmental pollutant that affects adversely a number of organs in humans and other mammals, including the kidneys, liver, lungs, pancreas, testis, and placenta. The liver and kidneys, which are the primary organs involved in the elimination of systemic Cd, are especially sensitive to the toxic effects of Cd. Because Cd ions possess a high affinity for sulfhydryl groups and thiolate anions, the cellular and molecular mechanisms involved in the handling and toxicity of Cd in target organs can be defined largely by the molecular interactions that occur between Cd ions and various sulfhydryl-containing molecules that are present in both the intracellular and extracellular compartments. A great deal of scientific data have been collected over the years to better define the toxic effects of Cd in the primary target organs. Notwithstanding all of the new developments made and information gathered, it is surprising that very little is known about the cellular and molecular mechanisms involved in the uptake, retention, and elimination of Cd in target epithelial cells. Therefore, the primary purpose of this review is to summarize and put into perspective some of the more salient current findings, assertions, and hypotheses pertaining to the transport and handling of Cd in the epithelial cells of target organs. Particular attention has been placed on the molecular mechanisms involved in the absorption, retention, and secretion of Cd in small intestinal enterocytes, hepatocytes, and tubular epithelial cells lining both proximal and distal portions of the nephron. The purpose of this review is not only to provide a summary of published findings but also to provide speculations and testable hypotheses based on contemporary findings made in other areas of research, with the hope that they may promote and serve as the impetus for future investigations designed to define more precisely the cellular mechanisms involved in the transport and handling of Cd within the body.     

Influence of cadmium and copper on tissue element levels of pregnant rats

In the current study, we examined the effects of Cd on Cd, Cu, Zn and Fe levels in placenta and maternal and fetal plasma and tissues, the placental weight, total fetal and maternal body weights, and fetal and maternal tissue weights during pregnancy. A total of 21 adult female rats were treated during gestation with drinking water containing one of the following: 70 mg/L of CdCl₂, a combination of 70 mg/L of CdCl₂ and 70 mg/L of CuSO₄, or no addition (control). Placenta Cu and Fe levels, fetal liver and kidney Cu levels, and fetal liver tissue weights were lower in the group administered Cd than in the control group. Also, Cd levels in the placenta, maternal and fetal liver, and maternal kidney were higher in the group treated with Cd than in controls. In the group administered both Cd and Cu, fetal body and tissue weights did not change, but Cd levels in the placenta, maternal and fetal liver, and maternal kidneys were higher than in controls. Zn and Fe levels in the maternal kidney and fetal liver were also lower in this group. Cd exposure during pregnancy resulted in Cd accumulation in maternal and fetal tissues during pregnancy and a decrease in the total weight of fetuses, and the combination of Cd and Cu caused some changes in the both maternal and fetal levels of Cu, Zn, and Fe, but it did not cause changes in the total fetal body weight or the weights of individual tissues.     

Effects of DN-2327, a new anxiolytic,diazepam and buspirone on exploratory activity of the rat in an elevated plus-maze

In the present study, the effects of a new anxiolytic, DN-2327, were compared to those of diazepam and buspirone in rats in the elevated plus-maze test. Two indices of anxiety were obtained in this test: the number of entries into the open arms expressed as a percentage of the total number of arm entries and the percentage of time spent on the open arms. Both a typical anxiolytic, diazepam, at 2.5, 5 and 10 mg/kg, PO and a new anxiolytic, DN-2327, at 2.5 and 5 mg/kg, PO dose-dependently increased the two indices: the percentage of time spent on the open arms and the percentage of open-arm entries. On the other hand, pentylenetetrazol (PTZ) at 10 and 20 mg/kg, IP decreased the two indices dose dependently as did yohimbine at 1.5 and 3 mg/kg, IP. DN-2327 at 2.5 and 5 mg/kg, PO and diazepam at 5 and 10 mg/kg, PO dose dependently and significantly increased the two indices that were suppressed following administration of PTZ at 10 mg/kg, IP. The effects of both DN-2327, 5 mg/kg, PO, and diazepam, 10 mg/kg, PO, on the two indices were significantly antagonized by the benzodiazepine (BZD) receptor antagonist flumazenil, 20 mg/kg, IP. Buspirone (2.5–20 mg/kg, PO) did not affect either of the two responses but dose dependently decreased the number of rearings, although in the Vogel conflict test, the anti-conflict activity of buspirone was equipotent to that of diazepam and DN-2327 at the minimum effective dose (10 mg/kg, PO) of each drug. In conclusion, the present experiment revealed that the anxiolytic effect of DN-2327 in this test was clear, whereas buspirone showed no apparent effect.     

Subacute toxicity evaluation of a new camptothecin anticancer agent CKD-602 administered by intravenous injection to rats

The subacute toxicity of a new camptothecin anticancer agent, CKD-602, was investigated after 4-week repeated intravenous administration of the chemical in Sprague-Dawley rats. The test chemical was administered intravenously to rats at dose levels of 0, 0.003, 0.013, or 0.067 mg/kg/day for males and 0, 0.004, 0.018, or 0.089 mg/kg/day for females. At the end of the treatment period, 10 rats/sex/group were sacrificed. The remaining 5 rats/sex in the vehicle control and high dose groups continued the study without treatment for 2 weeks (recovery period). During the test period, clinical signs, mortality, body weights, food and water consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross findings, organ weights, and histopathology were examined. In both sexes of the high dose group, an increase in the incidence of abnormal clinical signs and paleness of the eyes, a reduction in the body weight gain, food consumption and urine protein, and an increase in the water consumption were observed. Hematological investigations revealed a decrease in the red blood cells, hemoglobin and hematocrit and an increase in the mean corpuscular volume, mean corpuscular hemoglobin, platelets, and reticulocytes in a dose-dependent manner. Serum total cholesterol and total protein values were lower in females than those of controls, but not in males. An increase in the heart and liver weights and a decrease in the thymus weight were also found. Histopathological alterations included an increase in the incidence of atrophy of the sternal marrow, atrophy, fibrosis and mast cell hyperplasia of the femoral marrow, atrophy of the white pulp and extramedullary hematopoiesis of the spleen, atrophy of the thymus, auricular hypertrophy of the heart, extramedullary hematopoiesis and centriacinar telangiectasis of the liver, follicular degeneration of the ovary, and inflammation of the tail. The major treatment-related effects were not recovered at the end of 2-week recovery period. There were no adverse effects in the low and middle dose groups of both genders. In the present experimental conditions, the target organs were determined to be bone marrow, blood cells, spleen, liver, thymus, and heart. The no-observed-adverse-effect level was considered to be 0.013 mg/kg/day for males and 0.018 mg/kg/day for females.     

The concept of conditional pharmacology and toxicology

The concept of conditional pharmacology as initially elucidated by Dr Michael Whitehouse and his colleagues from their studies of drug-disease interactions has broad import in a rational drug discovery and development programme. The concept can be extended to toxicology and thus can be viewed as encompassing virtually all means and methods of discovering and enhancing the efficacy, while reducing the toxicity of drugs and biologics. The concept involves employing the physiological or metabolic activity, genetic and/or molecular structure of the host, of the disease process and/or of the parasite to activate and target the drug or biologic, as well as to regulate and delimit its activity. Thus, the concept not only applies to the treatment of inflammatory diseases, but also to the treatment of neoplastic and infectious diseases, to facilitating wound healing, and is in fact an underlying assumption, and expected consequence, of successful gene therapy. The concept applies to clinical studies as well, arguing for more pharmacokinetic and chrono-pharmacological studies in the early phases of clinical testing and the inclusion in later-stage clinical trials of more diverse populations, as regards age, gender and ethnicity, if the indication warrants. Facilitating and monitoring compliance, post-as well as pre-market approval, also are critical components of the fully implemented concept.     

北方旱寒区冬油菜根系抗寒指标分析

为了研究北方旱寒区冬油菜根部生理特性与其抗寒性的相关性,对6个不同抗寒性冬油菜品种整个生育期根部的干物质积累、含水量、可溶性蛋白含量及相关保护性酶活性的变化进行了分析,并将各指标与越冬率进行了相关性分析,用与越冬率相关性显著、极显著的6个指标对冬油菜进行聚类分析。结果表明,降温前,各指标与越冬率的相关性均不显著;降温后,越冬率与可溶性蛋白含量、SOD活性极显著正相关,与POD、APX活性显著正相关,与根直径、根冠比、CAT活性正相关,与根含水量负相关;根据聚类分析结果,可将试验材料分为3类,第1类：07-G01,天油2号,天油4号,此类抗寒性弱,为耐寒性品种;第2类：上党,74-1,此类抗寒性一般,为中抗寒性品种;第3类：陇油6号,此类抗寒性强,为强抗寒性品种。由此推断,在北方旱寒区,冬油菜越冬前根直径大、根冠比高、根含水量低有利于冬油菜安全越冬;降温后,冬油菜可溶性蛋白、SOD、POD 、CAT和 APX酶活性值越高,其抗寒性越强。