Characterization of rat somatotroph and mammotroph secretory granules. Presence of sulfated molecules

The contents of a purified somatotroph and mammotroph secretory granule fraction isolated from rat anterior pituitaries were solubilized in 4 M urea and analyzed by PAGE. In gels electrophoresed under a variety of conditions and stained with Coomassie Blue only two major bands, identified as GH and PRL, were present. In gels stained with Stains-All (which stains anionic substances), several additional bands were detected. When quarter pituitaries were labeled with a [3H]amino acid mixture, GH and PRL accounted for greater than 95% of the radioactivity incorporated into the granules. After labeling with [35S]sulfate, two classes of radiolabeled sulfated components were detected in the granules: a class of trypsin-sensitive macromolecular components which were coincident with two of the bands seen after Stains-All, and a class of low molecular weight components. In order to examine the distribution of the two classes of sulfated components within somatotroph and mammotroph granules, granules were suspended in 0.4% (w/v) Lubrol PX at pH 4.0, a treatment which has been shown to selectively solubilize somatotroph granule contents leaving mammotroph granule cores intact. This treatment was found to solubilize greater than 95% of the GH and greater than 99% of the radiolabeled, low molecular weight sulfated components; in contrast, there was virtually no solubilization of either PRL or macromolecular sulfated components. The findings indicate (a) that [35S]sulfate is incorporated into both somatotroph and mammotroph granules, and (b) that the low molecular weight sulfated components are associated with somatotroph granules whereas the macromolecular sulfated components are associated with mammotroph granules.     

养血清脑颗粒联合氟哌噻吨美利曲辛治疗紧张性头痛临床疗效观察

目的：研究养血清脑颗粒联合氟哌噻吨美利曲辛治疗紧张性头痛的临床疗效。方法：将2011年12月-2014年2月本院门诊收治的68例紧张性头痛患者给予养血清脑颗粒联合氟哌噻吨美利曲辛治疗,观察治疗前后的头痛程度、负面情绪及生活质量。结果：治疗后患者头痛程度及负面情绪评分明显低于治疗前,生活质量明显高于治疗前;患者的疼痛数字量表（NRS）评分为（1.4±0.2）分、汉密尔顿焦虑量表（HAMA）评分为（13.6±1.5）分、汉密尔顿抑郁量表（HAMD）评分为（13.8±2.6）分,均明显低于治疗前;躯体功能（72.8±9.6）分、心理功能（59.5±5.6）分、社会功能（57.7±2.87）分、总体生活质量（81.3±8.8）分,均明显高于未治疗前（P〈0.05）。结论：养血清脑颗粒联合氟哌噻吨美利曲辛有助于缓解头痛和负面情绪,提高患者的疼痛耐受性,对治疗紧张性头痛具有重要意义。     

参地颗粒对慢性肾炎脾肾亏虚证患者血清 VEGF、尿 EGF 的干预作用

目的：观察慢性肾小球肾炎（ CGN）脾肾亏虚证患者血清血管内皮生长因子（ VEGF）和尿表皮生长因子（EGF）的变化及参地颗粒的干预作用。方法：64例 CGN 脾肾亏虚证患者随机分为对照组和治疗组各32例,实际完成61例（对照组31例,治疗组30例）,并设正常组20例。治疗组口服参地颗粒,对照组口服氯沙坦钾片,疗程均为8周,观察治疗前后临床疗效、24 h 尿蛋白定量、尿红细胞计数、血清 VEGF 和尿 EGF 水平变化,并与正常组比较。结果：治疗组临床疾病总有效率和中医证候疗效总有效率均为86．67%,优于对照组的61．30%（P 〈0．05）;治疗组治疗后24 h 尿蛋白定量、尿红细胞计数均较治疗前降低（P 〈0．05）,而对照组治疗后24 h 尿蛋白定量较治疗前降低（P 〈0．05）,但尿红细胞计数降低不明显（P 〉0．05）;两组患者治疗前血清 VEGF 和尿 EGF 水平明显高于正常组（P 〈0．01）,治疗后两组血清 VEGF 和尿 EGF 水平均下降（P 〈0．05）,且治疗组优于对照组（P 〈0．05）。结论：CGN 脾肾亏虚证患者血清 VEGF、尿 EGF 水平上升,参地颗粒可显著降低患者血清 VEGF、尿 EGF 水平,降低中医证候积分,改善临床症状,其机制之一可能与其降低患者血清 VEGF、尿 EGF 水平有关。     

温阳解毒化瘀颗粒对肠源性内毒素血症大鼠肠黏膜上皮紧密连接的影响

目的：研究温阳解毒化瘀颗粒对肠源性内毒素血症（ IETM ）模型大鼠结肠黏膜上皮紧密连接的影响,探索其抗肝衰竭的作用机制。方法：将大鼠随机分为正常组、模型组、温阳解毒化瘀颗粒（实验组）和对照组4组,采用D-半乳糖胺（D-gal）腹腔注射致肝衰竭ITEM大鼠模型。正常组在腹腔注射生理盐水24h后处死,模型组、实验组、对照组分别于造模后24h、48h、72h各取6只、7只、7只大鼠处死,检测各组肝功能、内毒素、结肠黏膜上皮咬合蛋白（occludin）及肌球蛋白轻链激酶（MLCK）。结果：模型组血清ALT/AST、内毒素、 MLCK表达水平均高于正常组, occludin表达低于模型组（ P＜0.01）;实验组血清ALT/AST、内毒素、 MLCK表达水平均低于模型组, occlu-din表达高于模型组（ P＜0.05）。结论：增强结肠粘膜上皮紧密连接功能,降低内毒素的吸收是温阳解毒化瘀颗粒抗肝衰竭的作用机制之一。     

Genetic variants associated with Hermansky-Pudlak syndrome

Abstract

Hermansky-Pudlak syndrome (HPS) is a rare autosomal recessive disorder characterized by defective biogenesis of lysosome-related organelles. Clinical manifestations include a bleeding diathesis due to a platelet delta storage pool deficiency, oculocutaneous albinism, inflammatory bowel disease, neutropenia, and pulmonary fibrosis. Ten genes associated with HPS are identified to date, and each gene encodes a protein subunit of either Biogenesis of Lysosome-related Organelles Complex (BLOC)-1, BLOC-2, BLOC-3, or the Adaptor Protein-3 complex. Several genetic variants and phenotypic heterogeneities are reported in individuals with HPS, who generally exhibit easy bruisability and increased bleeding. Desmopressin, pro-coagulants, or platelet transfusion may be used as prophylaxis or treatment for excessive bleeding in patients with HPS. However, response to desmopressin can be variable. Platelets are effective in preventing or treating bleeding in individuals with HPS, but platelets should be transfused judiciously to limit alloimmunization in patients with HPS who are at risk of developing pulmonary fibrosis and may be potential candidates for lung transplantation. The discovery of new genes associated with HPS in people with excessive bleeding and hypopigmentation of unknown etiology may be facilitated by the use of next-generation sequencing or panel-based genetic testing.     

Ubiquitin‐negative,eosinophilic neuronal cytoplasmic inclusions associated with stress granules and autophagy: An immunohistochemical investigation of two cases

Identification of the proteinaceous components of the pathological inclusions is an important step in understanding the associated disease mechanisms. We immunohistochemically examined two previously reported cases with eosinophilic neuronal cytoplasmic inclusions (NCIs) (case 1, Mori et al. Neuropathology 2010; 30: 648–53; case 2, Kojima et al. Acta Pathol Jpn 1990; 40: 785–91) using 67 antibodies against proteins related to cytoskeletal constituents, ubiquitin‐proteasome system, autophagy‐lysosome pathway and stress granule formation. Regional distribution pattern of eosinophilic NCIs in case 1 was substantially different from that in case 2. However, NCIs in both cases were immunonegative for ubiquitin and p62 and were immunopositive for stress granule markers as well as autophagy‐related proteins, including valosin‐containing protein. Considering that eukaryotic stress granules are cleared by autophagy and valosin‐containing protein function, our findings suggest that eosinophilic NCIs in the present two cases may represent the process of autophagic clearance of stress granules.     

中药配方颗粒自动调配系统调剂操作规程的标准化研究

探索建立完善的中药配方颗粒自动化调配系统调剂操作流程,使其规范化、标准化、自动化调配准确性高,以防差错和纠纷的发生。通过反复调剂操作,结合实际工作,将其在工作中发现的问题和突发情形进行归纳总结,并将实际经验制定成中药配方颗粒自动调配系统调剂操作规程。完善的操作流程,可保证药物颗粒的稳定性,有效杜绝颗粒先后应用顺序不一的情况;发现问题及时,纠正快。操作流程规范,方法得当,调配准确,避免差错。     

不同组织膜蛋白及二甲基亚砜对小脑颗粒细胞突起生长的影响

目的探讨不同组织膜蛋白提取物和二甲基亚砜对大鼠小脑颗粒细胞（CGC）突起生长的影响。方法提取成年大鼠肝、坐骨神经和脑白质的组织膜蛋白包被盖片,接种CGC于盖片上,同时将二甲基亚砜（DMSO）作为添加剂加入培养液,观察CGC突起的生长。结果①脑白质膜蛋白能明显抑制CGC突起的生长,随浓度增加抑制效应更加明显;坐骨神经膜蛋白对CGC突起生长抑制不明显,肝组织膜蛋白则能促进突起生长;②随DMSO浓度升高CGC突起生长逐渐受到抑制,当DMSO浓度小于1％时,CGC突起长度与对照对照差别不大。结论脑白质膜蛋白对CGC突起生长有明显抑制作用,而DMSO添加到培养液中在低浓度时不会显著影响神经元突起生长,可用于检测药物及疏水性分子对体外培养的神经细胞突起生长的作用。     

环鸟苷酸依赖性蛋白激酶拮抗酒精性中枢神经细胞毒性机制研究

目的 探讨环鸟苷酸依赖性蛋白激酶对酒精致鼠小脑颗粒细胞(cerebellar granule cells,CGC)毒性作用的影响机制.方法 通过光学显微镜观察一氧化氮(NO)释放剂,环鸟苷酸及环鸟苷酸依赖性蛋白激酶抑制剂对鼠小脑颗粒细胞存活的影响.结果 NO释放剂,环鸟苷酸在无酒精组中促进细胞的生存,并能对CGC产生保护作用,减少酒精对细胞的杀死.环鸟苷酸依赖性蛋白激酶抑制剂可抑制二者对细胞的作用.结论 环鸟苷酸依赖性蛋白激酶在NO释放剂和环鸟苷酸促鼠小脑颗粒细胞的存活和抵抗酒精性神经毒性过程中具有重要的作用.     

稳心颗粒配合胺碘酮治疗早搏的临床观察

目的:观察稳心颗粒治疗早搏的临床疗效。方法:将入选病例54例随机分为两组,治疗组(27例)口服稳心颗粒及胺碘酮片,对照组(27例)口服胺碘酮片,疗程均为4周。结果:总有效率治疗组为96.3%,对照组为85.19%,两组比较无统计学意义(P>0.05),胺碘酮片用量治疗组比对照组减少1/2,两组比较有统计学意义(P<0.05)。结论:稳心颗粒对心律失常有一定的治疗作用,并可减少胺碘酮的用量。