Isopycnic-zonal centrifugation of plasma membrane,sarcoplasmic reticular fragments,lysosomes, and cytoplasmic proteins from phasic skeletal muscle

Homogenates of the posterior latissimus dorsi muscle, a phasic muscle, were fractionated by a one-step zonal centrifugation technique into four major organelle populations and cytoplasmic constituents. These were: (1) Plasma membrane fragments with a modal equilibrium density of 1.10 and containing 5′-nucleotidase, alkaline phosphodiesterase, $\text{p-}\text{nitrophenylphosphatase}$ and acid phosphatase (β-glycerophosphate was used as the substrate). (2) Sarcoplasmic reticular fragments which could be further subdivided into calcium transport vesicles, with a modal equilibrium density of 1.16, that exhibited calcium uptake; K⁺-ATPase; leucyl-β-naphthylamidase; acid phosphodiesterase; acid phosphatase (using cytidine monophosphate as the substrate); and sarcoplasmic reticular lysosomes, with a modal equilibrium density of 1.18, possessing dipeptidyl-aminopeptidase II, cathepsin D, α-glucosidase, $\text{N-}\text{acetyl}\text{-β-}\text{glucosaminidase}$, and NADH oxidase activity. (3) Mitochondria with a modal equilibrium density of 1.21. (4) Catalase-containing vesicles with a modal equilibrium density of 1.22; and cytoplasmic constituents (modal density of 1.25) with phosphorylase, pyruvate kinase, myosin-ATPase, aldolase, and protein and RNA content. The purity of these organelles was equal to or better than previous efforts, with a 30-fold purification achieved for 5′-nucleotidase and alkaline phosphodiesterase. These results lend support to the hypothesis that the sarcoplasmic reticulum of phasic muscle, in addition to its specialized role in excitation-contraction coupling, represents a multifunctional membrane system, and that, similar to the smooth endoplasmic reticulum of other cells, it includes some membrane-bound lysosomal enzymes and NADH oxidase.     

Delta-sleep-inducing peptide (DSIP): An update

The isolation and characterization of delta-sleep-inducing peptide (DSIP) achieved from 1963 to 1977 were reviewed in 1984. The first reports describing sleep as well as extra-sleep effects of DSIP also were included in that work. Only two years later, much additional literature concerning DSIP has accumulated. Besides further sleep-inducing and/or-supporting effects of DSIP in animals, considerable work has been carried out to evaluate the potential use of the peptide for therapeutic purposes such as treatment of insomnia, pain, and withdrawal. Immunohistochemical as well as radioimmunochemical studies provided further insights into the natural occurrence of the nonapeptide and the distribution of DSIP-like material in the body, suggesting possible relations of the peptide to certain diseases. Various physiological functions of DSIP and a possible mechanism of action involving the modulation of adrenergic transmission remain to be established.     

Thioredoxin and HSP90α modulate ASK1–JNK1/2 signaling in stressed hepatocytes of Mugil cephalus

Induction of antioxidant proteins like thioredoxin (Trx) and heat shock protein 90α (HSP90α) is a crucial step in the cellular response to oxidative stress. Here, we report the impact of environmental stress on Trx and HSP90α expressions in freshly isolated hepatocytes of Mugil cephalus living in either a contaminated (Test; Ennore) or uncontaminated (Control; Kovalam) estuary. Modulation in the activities of signal transduction molecules like apoptosis signal-regulating kinase 1 (ASK1) and c-Jun NH₂-terminal kinase 1/2 (JNK1/2) were also investigated to understand their functional role under natural stressed condition. The expression pattern of the proteins was determined by immunoblotting and the relationship between the proteins was identified by regression analysis. Test fish hepatocytes demonstrated significant upregulation (P < 0.05) in the levels of Trx and HSP90α and insignificant inductions in the expression pattern of ASK1 and JNK1/2 than control fish hepatocytes. These findings provide direct evidence that Trx and HSP90α induction in fish hepatocytes under stress may aid cell survival by negatively regulating ASK1 expression and thereby functionally antagonizing the apoptotic role of JNK1/2 in natural aquatic systems.     

不同剂量舒芬太尼对心脏瓣膜置换术患者应激反应、炎性因子及心肌损伤的影响

目的:探讨不同剂量舒芬太尼对心脏瓣膜置换术患者应激反应、炎性因子及心肌损伤的影响。方法:根据随机数字表法将100例行心脏瓣膜置换术的患者分为低剂量组(n=33,舒芬太尼剂量为1.0μg/kg)、中剂量组(n=33,舒芬太尼剂量为1.5μg/kg)以及高剂量组(n=34,舒芬太尼剂量为2.0μg/kg),比较三组患者应激反应、炎性因子、心肌损伤等指标的变化以及围术期指标情况。结果:中剂量组、高剂量组麻醉诱导后(T1)、插管后1 min(T2)、插管后5 min(T3)、插管后10 min(T4)时间点心率(HR)、平均动脉压(MAP)均低于低剂量组同时间点,且高剂量组低于中剂量组(P<0.05)。与低剂量组比较,中剂量组、高剂量组阻断后30 min(T6)、开主动脉后2h(T7)以及术后1d(T8)时间点白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)均降低(P<0.05)。与低剂量组比较,中剂量组、高剂量组体外循环停机2h(T9)、体外循环停机8h(T10)、体外循环停机24h(T11)、体外循环停机48h(T12)时间点心肌肌钙蛋白I(cTnI)、肌酸磷酸激酶同工酶(CK-MB)均降低(P<0.05)。低剂量组、中剂量组重症监护室(ICU)滞留时间、拔管时间显著短于高剂量组(P<0.05),而三组心血管不良事件发生率比较差异无统计学意义(P>0.05)。结论:给予1.0μg/kg舒芬太尼麻醉的患者应激反应小,1.5μg/kg、2.0μg/kg舒芬太尼可更好地控制心脏瓣膜置换术患者炎性反应,同时对患者心肌损伤有一定的保护作用,但2.0μg/kg舒芬太尼会延长患者ICU滞留时间、拔管时间。     

右美托咪定复合罗哌卡因对膝关节置换术后镇痛镇静效果、睡眠质量及应激反应的影响

目的:探讨右美托咪定复合罗哌卡因对膝关节置换术后的临床效果。方法:选取2015年2月～2018年12月期间在武警陕西省总队医院择期行膝关节置换术患者103例,根据随机数字表将患者分成对照组(n=51)和研究组(n=52),对照组股神经阻滞时给予罗哌卡因,研究组在对照组基础上联合右美托咪定。比较两组患者镇痛镇静效果、睡眠质量、应激反应以及不良反应。结果:研究组术后12 h、术后24 h、术后48 h视觉模拟评分(VAS)低于对照组(P0.05),Ramsay镇静评分则高于对照组(P0.05)。研究组术后24 h、术后48 h阿森斯失眠量表(AIS)评分均低于术前,且呈先降低后升高趋势(P0.05)。研究组术后24 h、术后48 h AIS评分低于对照组(P0.05)。两组患者手术开始即刻、术毕的去甲肾上腺素(NE)、肾上腺素(E)水平均上升,且呈先升高后降低趋势(P0.05);研究组手术开始即刻、术毕的NE、E水平低于对照组(P0.05)。两组不良反应发生率对比无统计学差异(P0.05)。结论:右美托咪定复合罗哌卡因方案应用于膝关节置换术,镇痛镇静效果确切,可减轻术中应激反应,有效改善睡眠质量,且不增加不良反应发生率,临床应用价值较高。     

单切口腹腔镜在结直肠手术中的应用效果及对炎症因子与应激反应的影响

目的:探讨单切口腹腔镜在结直肠手术中的应用效果及对炎症因子与应激反应的影响。方法:选择2018年1月至2019年10月我院接诊的98例结直肠癌患者,通过随机数表法分为2组,每组49例。观察组使用单切口腹腔镜结直肠癌根治术,对照组使用传统腹腔镜结直肠癌根治术,常规5孔操作法。比较两组围术期相关情况、手术前后血清炎症因子水平的变化、手术后应激反应及并发症的发生情况。结果:两组淋巴结清扫个数、中转开腹例数、术后排气时间比较差异均无统计学意义(P0.05),观察组手术时间明显长于对照组,脐切口长度、住院时间、术中出血量均显著短于或低于对照组;两组术后1 d、3 d、5 d时高敏C反应蛋白(hs-CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、去甲肾上腺素(NE)、肾上腺素(E)、皮质醇(Cor)水平均明显高于术前(P0.05);且观察组术后1d、3d、5d时hs-CRP、TNF-α、IL-6、NE、E、Cor较对照组显著降低(P0.05);两组吻合口瘘、吻合口狭窄、吻合口出血、小肠穿孔、肠梗阻、感染的总发生率比较差异无统计学意义(P0.05)。结论:和传统5孔操作法相比,单切口腹腔镜结直肠癌根治术治疗结直肠癌患者的手术时间更长,但术后炎症因子表达更低,应激反应更小,有利于患者术后恢复。     

Natural roles of nonribosomal peptide metabolites in fungi

Over 90 years have passed since Alexander Fleming's discovery of penicillin, the first recognized, naturally occurring antibiotic. Penicillin is a representative of a group of metabolites produced by large multienzyme complexes [nonribosomal peptide synthetases (NRPSs)] in a ribosome-independent fashion. Nonribosomal peptides (NRPs) are structurally diverse metabolites produced almost exclusively by bacteria and fungi. NRPs include bioactive compounds useful for pharmaceutical applications (e.g., antibiotics, antitumor compounds, and immunosuppressants) and therefore much progress has been made in our understanding of medically relevant characteristics of NRPs in the past decades. Natural roles of NRP metabolites, on the other hand, have been largely ignored, and much less is known about the biological/physiological significance of NRPs under natural settings. In the present review, we summarize past and current work on natural functions of NRPs in their fungal producers, with a focus on virulence, development, and stress tolerance, and highlight the diverse roles these small peptide metabolites play. Some NRPs are involved in interactions with host organisms, others work in fungus-environment interfaces, and still others are crucial for vegetative and reproductive development of the producing fungi.     

结核分枝杆菌小RNA及其作用的研究进展

细菌小RNA是一类长度在50～500个核苷酸之间的不具有编码蛋白质功能,但具有转录后调控作用的RNA,在细菌中参与调控细菌多种生理和病理活动,如调节细菌代谢和毒力作用等过程。近年来,在结核分枝杆菌已经鉴定出近200种小RNA,并证明这些小RNA参与结核分枝杆菌的生理和病理过程。本文对结核分枝杆菌小RNA在细菌生长繁殖、毒力因子调控、细菌耐药和巨噬细胞内应激环境的适应等方面的作用进行综述。