外用药物制剂药品生产质量管理规范实施现状分析与研究

本文从外用药物制剂实施现状出发,通过在数据调研和大量文献收集的基础上,对从药品生产质量管理规范(GMP)的发展及其在我国的推行历史角度,分析并阐述了现阶段外用药物制剂GMP的实施状况,并对某制药集团属下25家药企实施2010版GMP的现状进行调查分析。由于历史发展的原因促使药品管理尚未完全规范以外,GMP标准的过时性和落后性影响了我国GMP工作的深入实施。只有保证了日常的GMP实施,才能有效地保证药品生产的高质量、低风险,有效地促进药品生产的质量保障,使药品真正达到国际水平,走向现代化、国际化。     

一种动物视网膜压力仪的设计

目的:缺血性视网膜损伤是眼科常见的一种临床表现,对其进行研究需要建立缺血性视网膜损伤模型。采用生理盐水前房加压灌注法制作高眼压模型是最为常用的一种造模方法,但目前国内尚未有这种专用的模型制作装置。针对这种情况,我们设计了一种动物视网膜压力仪,用于制备缺血性视网膜损伤模型。方法:该装置采用压缩空气作为压力源,以气-液隔离器为施压执行器,通过压力传感器检测压力,用控制器控制压力和加压时间,并且控制器采用模块化设计方法进行设计,其压力的控制采用通用的智能工业调节器来实现,时间控制采用通用的智能时间继电器来实现。结果:该装置可自动按设定的时间对眼前房施加压力,并且时间准确,压力恒定。结论:该装置压力稳定,定时准确,可根据实验需要设定压力的大小和定时时间。预先设定压力和定时时间后,整个实验过程自动进行。使用该装置不仅保证了实验过程的一致性,使实验操作更加规范和严谨,而且提高了工作效率。该装置性能可靠,结构简单,制作容易,操作简单,使用方便。     

中性粒细胞弹性蛋白酶的过表达促进K562细胞增殖并抑制其凋亡

目的利用Ad Easy系统构建携带人中性粒细胞弹性蛋白酶(NE)基因的重组腺病毒表达质粒,并感染K562细胞,观察其对K562细胞增殖和凋亡的影响。方法以急性早幼粒细胞白血病(APL)细胞株HL-60的RNA为模板,经反转录PCR得到NE基因的编码区并克隆入穿梭载体p Ad Track-CMV,得到p Ad-NE,经HindⅢ、EcoRⅤ酶切鉴定和测序正确后,将其转化入p Ad Easy-1-BJ5183进行同源重组,得到重组子Ad-NE,经PacⅠ酶切线性化后转染AD293细胞进行包装。14 d之后收获原代病毒,经5轮扩增后,测定病毒滴度,进行PCR鉴定。感染K562细胞后,荧光显微镜初步观察感染效率,同时用流式细胞术检测感染效率;Western blot法进一步鉴定NE是否表达上调;CCK-8法测定细胞增殖情况;流式细胞术测定细胞凋亡和细胞周期变化。结果酶切鉴定及测序结果表明重组穿梭质粒p Ad-NE构建成功;酶切鉴定显示同源重组成功,得到重组子Ad-NE;荧光显微镜表明病毒包装成功;经过5轮扩增后,病毒滴度达到1.64×1012pfu/m L;感染K562细胞后,荧光显微镜观察感染效率达80%左右;Western blot法检测到NE表达上调;CCK-8实验显示NE表达上调后K562细胞增殖能力增强;流式细胞术显示S期细胞明显增多并且细胞凋亡减少。结论过表达NE可促进K562细胞增殖并抑制其凋亡。     

Gene therapy blockade of dorsal striatal p11 improves motor function and dyskinesia in parkinsonian mice

Complications of dopamine replacement for Parkinson’s disease (PD) can limit therapeutic options, leading to interest in identifying novel pathways that can be exploited to improve treatment. p11 (S100A10) is a cellular scaffold protein that binds to and potentiates the activity of various ion channels and neurotransmitter receptors. We have previously reported that p11 can influence ventral striatal function in models of depression and drug addiction, and thus we hypothesized that dorsal striatal p11 might mediate motor function and drug responses in parkinsonian mice. To focally inhibit p11 expression in the dorsal striatum, we injected an adeno-associated virus (AAV) vector producing a short hairpin RNA (AAV.sh.p11). This intervention reduced the impairment in motor function on forced tasks, such as rotarod and treadmill tests, caused by substantia nigra lesioning in mice. Measures of spontaneous movement and gait in an open-field test declined as expected in control lesioned mice, whereas AAV.sh.p11 mice remained at or near normal baseline. Mice with unilateral lesions were then challenged with l-dopa (levodopa) and various dopamine receptor agonists, and resulting rotational behaviors were significantly reduced after ipsilateral inhibition of dorsal striatal p11 expression. Finally, p11 knockdown in the dorsal striatum dramatically reduced l-dopa–induced abnormal involuntary movements compared with control mice. These data indicate that focal inhibition of p11 action in the dorsal striatum could be a promising PD therapeutic target to improve motor function while reducing l-dopa–induced dyskinesias.Pharmacologic replacement of depleted dopamine is the primary therapeutic approach to treating Parkinson’s disease (PD). Although this usually improves the major motor problems of this disorder, complications of medical therapy can often limit both dosing and effectiveness. Among the most common adverse effects limiting dopamine replacement therapy for PD is the development of abnormal involuntary movements (AIMs), also known as levodopa-induced dyskinesia (LID) (1). Treatment of LID usually requires reducing the dosage of dopaminergic medications to below the threshold for major complications, although certain pharmacotherapies or surgeries can improve LID as well (1). Understanding both the anatomic location and molecular pathways underlying dyskinesia responses to dopamine replacement therapy is necessary to develop improved therapies, which can reduce motor symptoms without this debilitating problem.Previous studies have identified certain signaling pathways that may influence the development of dyskinesia. The primary site of action of l-dopa (levodopa) on PD motor symptoms after conversion to dopamine is the dorsal striatum, owing to the loss of the normal dopaminergic inputs from the substantia nigra pars compacta (2). This same region has also been shown to be responsible for motor complications of l-dopa therapy, including LID. Specifically, neurons harboring the D1 dopamine receptor appear to be primarily involved in these responses (3–5). Furthermore, other signaling pathways, including the serotonin 5-HT1B receptor, seem to modulate the response of these neurons to dopamine replacement therapy (6, 7). Nonetheless, it has been difficult to identify potential therapeutic targets that both improve motor function and reduce dyskinesia.Here we demonstrate that dorsal striatal p11 is a key regulator of dopamine responses in PD. We previously reported that p11, a small adaptor protein also known as S100A10, binds to specific serotonin receptor subtypes, including 5-HT1B (8–10). Because activation of the 5-HT1B serotonin receptor (5-HT1BR) reduces dyskinesia, and p11 binds to 5-HT1BR and potentiates 5-HT1B activity, we hypothesized that dorsal striatal p11 may influence the response to dopamine replacement therapy. We found that inhibition of p11 expression in the dorsal striatum improved motor function in parkinsonian mice. Surprisingly, blockade of dorsal striatal p11 expression profoundly inhibited dyskinesias in response to chronic l-dopa treatment, to a greater extent than pharmacologic activation of 5-HT1B in controls. This indicates that inhibition of striatal p11 is a promising potential target to block dyskinesias while improving motor function in PD, and that these effects likely occur through a mechanism other than 5-HT1B.     

山东省农村学龄前儿童心理健康状况及影响因素

目的：了解山东省农村学龄前儿童心理健康现状和影响因素,为进一步干预提供理论依据。方法采用自行设计儿童基本情况调查表与长处和困难问卷（SDQ）家长版对699名3～5岁农村学龄前儿童进行调查分析。结果被调查儿童情绪和行为问题检出率为29．76％,各因子异常检出率分别为：情绪症状,4．33％;品行问题,9．76％;多动注意力不集中,11．12％;同伴交往问题,6．16％;亲社会行为,12．85％。男童品行问题检出率高于女童（χ2＝14．38,P＜0．05）,留守儿童情绪和行为问题检出率高于非留守儿童（χ2＝4．07,P＜0．05）。Logistic回归分析显示,监护人不知晓健康的生活方式、对抚养孩子感到有负担和精力不足、在批评孩子前不给其申诉的机会以及孩子边吃饭边看电视是儿童出现情绪和行为问题的危险因素。结论农村学龄前儿童情绪和行为问题检出率较高,应当引起重视并进行有效的干预。     

环丙沙星在机械通气下呼吸道重症感染患者中的肺组织渗透性研究

目的评价环丙沙星在机械通气的下呼吸道重症感染患者的药代动力学和肺组织渗透性研究。方法入选用有创通气治疗的下呼吸道重症感染患者12例,静脉滴注环丙沙星0.2 g或0.4 g,给药间隔为12 h或24 h,给药后3 h进行支气管镜和肺泡灌洗,治疗72 h后开始留取血清标本,用高效液相色谱(HPLC)法检测血清和肺泡灌洗的药品浓度。结果共检测环丙沙星血清药品浓度84份,肺泡灌洗药品浓度12份。环丙沙星0.2 g患者Cmax和给药3 h的肺泡上皮衬液浓度分别为(4233.4±1165.9)和(1202.5±291.5)ng·m L-1,肺组织渗透率为(0.72±0.54)。环丙沙星0.4 g患者Cmax和给药3 h的肺泡上皮衬液浓度分别为(5342.7±1210.2)和(2219.4±612.7)ng·m L-1,肺渗透率为(0.90±0.67)。环丙沙星0.4 g的肺组织渗透率显著高于0.2 g(P<0.05)。结论环丙沙星在有创机械通气的下呼吸道重症感染患者有较好的肺组织渗透性,且提高单次使用剂量可明显增强感染患者肺组织渗透性。     

SD大鼠生长发育期心脏结构及功能的超声心动图评价

目的应用高频超声仪动态观测SD雄性大鼠生长发育阶段心脏结构和功能变化规律，为发育期实验大鼠动物模型制备及功能评价提供依据。方法利用小动物超声仪连续观测SD雄性大鼠出生后1—9周左心室结构及功能的动态变化。结果（1）M超检测结果显示左室壁厚度及左室内径随生长发育而增加，超声评估左心室重量与解剖称重结果高度相关（r=0．9326，P〈0．0001）；（2）左室短轴缩短率（FS％）在发育期无显著变化；（3）脉冲多普勒测定舒张早期峰值血流速度（E峰）与舒张晚期峰值血流速度（A峰）之比（E／A）〉1，在发育过程中无显著性变化；（4）组织多普勒检测提示：左室后壁心肌的舒张早期（Em）、舒张晚期（Am）及收缩期（Sm）运动速度出生后1—6周逐渐增加，6周时达平台期。结论高频小动物超声仪经胸超声心动图所检测的心脏结构能准确地反映大鼠心脏生长发育变化趋势；心脏整体收缩及舒张功能在出生后1周即达稳定状态；组织多普勒检测左室后壁心肌纵轴运动在出生后逐渐成熟完善，于6周达到稳定状态。     

高龄和年轻冠心病患者冠状动脉介入治疗临床对比分析

目的探讨高龄和年轻冠心病患者冠状动脉介入治疗效果。方法选择在我科行冠状动脉介入治疗的冠心病患者320例,根据年龄分为年轻组(146例)、高龄组(62例)和对照组(112例),分析各组患者临床特点、病变特点及介入治疗效果。结果以急性心肌梗死起病的患者在年轻组、高龄组和对照组分别占45.2%,17.7%和23.2%(P=0.000);以心绞痛起病的患者中1周内发生心肌梗死的患者分别占47.2%、14.3%和40.0%(P=0.018);非ST段抬高心肌梗死的发生率分别为1.9%、10.5%和1.9%(P=0.000);高血压患者在3组中分别占40.4%、67.7%和56.3%(P=0.001);吸烟患者分别占71.2%、22.6%和55.4%(P=0.000);肾功能不全患者分别占1.4%、16.1%和2.7%(P=0.000);住院病死率分别占0、6.5%和0.9%(P=0.003)。结论年轻患者多以急性心肌梗死起病;高龄患者病变复杂,合并多系统疾病多,严重并发症发生率偏高。     

体外心肺复苏院内急救流程的优化及效果评价

目的 通过对体外心肺复苏院内急救流程（以下简称“急救流程”）的优化,缩短抢救时间,减少不良事件的发生。方法 2018年10月成立医疗失效模式与效应分析管理团队,应用医疗失效模式与效应分析法分析急救流程缺陷,通过文献检索、专家会议法、专家咨询法优化急救流程并于2019年1月应用于临床。临床应用急救流程2年后分析比较急救流程优化前后风险优先指数、各环节用时及不良事件发生情况。结果 急救流程优化后急救流程风险优先指数由（217.63±20.60）分降低到（102.73±39.95）分,各环节用时缩短,各类不良事件发生率均有所下降,差异均具有统计学意义（P<0.05）。结论 应用医疗失效模式与效应分析法优化急救流程,可以有效改善急救延时,减少不良事件的发生。