不同麻醉方式对老年糖尿病患者全膝置换术后糖代谢的影响

目的:探讨不同麻醉方式对老年糖尿病患者全膝置换术(total knee arthoplasty,TKA)后糖代谢的影响。方法:2018年2月到2020年3月选择在本院进行择期TKA的老年糖尿病合并膝关节骨性关节炎患者88例,随机原则分为研究组与对照组,各44例。对照组给予常规静吸复合全身麻醉,研究组在对照组麻醉的基础上给予复合股神经阻滞,检测两组术后空腹血糖值,并记录两组膝关节功能改善情况。结果:研究组术后1 d、3 d与7 d静息状态下和活动状态下的疼痛视觉模拟评分法(Visual Analogue Scale/Score,VAS)评分都显著低于对照组(P<0.05)。研究组术后1 d、3 d与7 d的空腹血糖值都显著低于对照组(P<0.05)。研究组术后1 d、3 d与7 d的患肢膝关节活动度都显著高于对照组(P<0.05)。结论:股神经阻滞联合全麻在老年糖尿病患者TKA中的应用能缓解术后疼痛,减低术后血糖水平,并且促进提高膝关节活动度有益预后。     

超声引导下硬膜外阻滞在老年髋关节置换手术中的应用

目的:探讨超声引导下硬膜外阻滞在老年髋关节置换手术中的应用方法与效果。方法:2017年6月至2020年5月选择在本院进行髋关节置换手术的老年患者112例,根据随机数字表法把患者分为研究组与对照组,各56例。研究组给予超声引导下硬膜外阻滞,对照组给予传统的静脉持续镇痛。两组都给予全麻诱导与维持,记录镇痛效果与患者术后康复情况。结果:两组的性别、年龄、麻醉时间、手术时间与术中出血量等对比差异无统计学意义(P>0.05),研究组的术后住院时间显著短于对照组(P<0.05)。两组术后1 d、3 d、7 d的疼痛视觉模拟评分法(Visual Analogue Scale/Score,VAS)评分都低于术前1 d,观察组也都显著低于对照组,对比差异都有统计学意义(P<0.05)。研究组术后1 d、3 d、7 d的髋关节活动度都显著高于对照组(P<0.05)。研究组术后1 d、3 d、7 d的血清P物质(Substance P,SP)、前列腺素E2(Prostaglandin E2,PGE2)含量都高于术前1 d,观察组低于对照组,对比差异都有统计学意义(P<0.05)。结论:超声引导下硬膜外阻滞在老年髋关节置换手术中的应用能抑制血清SP、PGE2的释放,能缓解患者术后疼痛,促进髋关节功能的恢复,缩短患者的康复时间。     

A focus on fixation

Three fixation issues related to immunostaining are discussed here: 1) Generally, a tissue block is fixed, then embedded and sectioned (pre-fixation). The type of fixative applied, crosslinking or coagulating, has an impact on selecting an epitope retrieval method. Individual antigens have a fixation–retrieval characteristic. 2) A long fixation time, especially with crosslinking fixatives, may compromise the result of immunostaining. This negative effect varies among different antigens and can be partially restored by applying a more sensitive/efficient detection system such as tyramide amplification. 3) Sections cut from a fresh frozen tissue block usually are acetone fixed (post-fixation). This was accepted as the “gold standard” for a long time. Post-fixation, however, may have serious consequences for preservation of small peptides leaking from the cut open cells, whereas this is not the case with pre-fixed intact cells. Consequently, the concept of an acetone post-fixed cryostat tissue section as “gold standard” no longer exists and a more appropriate use of the terms immunohistochemistry and immunocytochemistry therefore seems justified. For many antibodies, it is not known whether a formalin fixed, paraffin embedded tissue specimen is appropriate. Suggestions are made for creating a positive control cell block for testing such antibodies.     

Mitotic chromosome size scaling in Xenopus

As rapid divisions without growth generate progressively smaller cells within an embryo, mitotic chromosomes must also decrease in size to permit their proper segregation, but this scaling phenomenon is poorly understood. We demonstrated previously that nuclear and spindle size scale between egg extracts of the related frog species Xenopus tropicalis and Xenopus laevis, but show here that dimensions of isolated mitotic sperm chromosomes do not differ. This is consistent with the hypothesis that chromosome scaling does not occur in early embryonic development when cell and spindles sizes are large and anaphase B segregates chromosomes long distances. To recapitulate chromosome scaling during development, we combined nuclei isolated from different stage Xenopus laevis embryos with metaphase-arrested egg extracts. Mitotic chromosomes derived from nuclei of cleaving embryos through the blastula stage were similar in size to replicated sperm chromosomes, but decreased in area approximately 50% by the neurula stage, reproducing the trend in size changes observed in fixed embryos. Allowing G2 nuclei to swell in interphase prior to mitotic condensation did not increase mitotic chromosome size, but progression through a full cell cycle in egg extract did, suggesting that epigenetic mechanisms determining chromosome size can be altered during DNA replication. Comparison of different sized mitotic chromosomes assembled in vitro provides a tractable system to elucidate underlying molecular mechanisms.     

Asp residues of the Glu-Glu-Asp-Asp pore filter contribute to ion permeation and selectivity of the Cav3.2 T-type channel

Voltage-activated Ca²⁺ channels are membrane protein machinery performing selective permeation of external calcium ions. The main Ca²⁺ selective filters of all high-voltage-activated Ca²⁺ channel isoforms are commonly composed of four Glu residues (EEEE), while those of low-voltage-activated T-type Ca²⁺ channel isoforms are made up of two Glu and two Asp residues (EEDD). We here investigate how the Asp residues at the pore loops of domains III and IV affect biophysical properties of the Ca_v3.2 channel. Electrophysiological characterization of the pore mutant channels in which the pore Asp residue(s) were replaced with Glu, showed that both Asp residues critically control the biophysical properties of Ca_v3.2, including relative permeability between Ba²⁺ and Ca²⁺, anomalous mole fraction effect (AMFE), voltage dependency of channel activation, Cd²⁺ blocking sensitivity, and pH effects, in distinctive ways.     

Regulation of a Dynamic Interaction between Two Microtubule-binding Proteins,EB1 and TIP150, by the Mitotic p300/CBP-associated Factor (PCAF) Orchestrates Kinetochore Microtubule Plasticity and Chromosome Stability during Mitosis

The microtubule cytoskeleton network orchestrates cellular dynamics and chromosome stability in mitosis. Although tubulin acetylation is essential for cellular plasticity, it has remained elusive how kinetochore microtubule plus-end dynamics are regulated by p300/CBP-associated factor (PCAF) acetylation in mitosis. Here, we demonstrate that the plus-end tracking protein, TIP150, regulates dynamic kinetochore-microtubule attachments by promoting the stability of spindle microtubule plus-ends. Suppression of TIP150 by siRNA results in metaphase alignment delays and perturbations in chromosome biorientation. TIP150 is a tetramer that binds an end-binding protein (EB1) dimer through the C-terminal domains, and overexpression of the C-terminal TIP150 or disruption of the TIP150-EB1 interface by a membrane-permeable peptide perturbs chromosome segregation. Acetylation of EB1-PCAF regulates the TIP150 interaction, and persistent acetylation perturbs EB1-TIP150 interaction and accurate metaphase alignment, resulting in spindle checkpoint activation. Suppression of the mitotic checkpoint serine/threonine protein kinase, BubR1, overrides mitotic arrest induced by impaired EB1-TIP150 interaction, but cells exhibit whole chromosome aneuploidy. Thus, the results identify a mechanism by which the TIP150-EB1 interaction governs kinetochore microtubule plus-end plasticity and establish that the temporal control of the TIP150-EB1 interaction by PCAF acetylation ensures chromosome stability in mitosis.     

Aberrant spindle dynamics and cytokinesis in Dictyostelium discoideum cells that lack glycogen synthase kinase 3

Eukaryotic cell division requires the co-ordinated assembly and disassembly of the mitotic spindle, accurate chromosome segregation and temporal control of cytokinesis to generate two daughter cells. While the absolute details of these processes differ between organisms, there are evolutionarily conserved core components common to all eukaryotic cells, whose identification will reveal the key processes that control cell division. Glycogen synthase kinase 3 (GSK-3) is a major protein kinase found throughout the eukaryotes and regulates many processes, including cell differentiation, growth, motility and apoptosis. In animals, GSK-3 associates with mitotic spindles and its inhibition causes mis-regulation of chromosome segregation. Two suppressor screens in yeast point to a more general effect of GSK-3 on cell division, however the direct role of GSK-3 in control of mitosis has not been explored outside the animal kingdom. Here we report that the Dictyostelium discoideum GSK-3 orthologue, GskA, associates with the mitotic spindle during cell division, as seen for its mammalian counterparts. Dictyostelium possesses only a single GSK-3 gene that can be deleted to eliminate all GSK-3 activity. We found that gskA-null mutants failed to elongate their mitotic spindle and were unable to divide in shaking culture, but have no chromosome segregation defect. These results suggest further conservation for the role of GSK-3 in the regulation of spindle dynamics during mitosis, but also reveal differences in the mechanisms ensuring accurate chromosome segregation.     

混合应用罗哌卡因、氯胺酮在新生儿骶管阻滞中的作用

目的:探明罗哌卡因、氯胺酮混合液在新生儿骶管阻滞术后镇痛的疗效和不良反应.方法:选取45例即将行骶管阻滞的新生儿,随机分成A、B、C三组,每组15人,其中A组患者行罗哌卡因骶管阻滞,B组患者行罗哌卡因骶管阻滞并氯胺酮皮下注射,C组患者行罗哌卡因、氯胺酮混合液骶管阻滞.术后于1、2、4、6、8、12、24h分别记录各患者的镇痛情况以及镇痛时间,采用FLACC法评价镇痛效果,以评分＜4分的维持时间作为有效镇痛时间.并且观察术后48h内不良反应的发生情况.结果:A组与B组之间比较镇痛时间以及术后不同时间点不需要镇痛的患者例数,发现两组间并没有较大差别(P＞0.05).相比较于A、B两组,C组患者阵痛时间明显延长,并且术后4、6、8、12、24h患者不需要镇痛明显增多(P＜0.05).而在不良反应方面,三组患者均未表现出过强的不良反应.结论:罗哌卡因、氯胺酮混合液用于新生儿骶管阻滞效果较佳,且不良反应少.     

WEE1 激酶及其抑制剂在抗肿瘤治疗中的应用进展

WEE1激酶是一种细胞周期调节蛋白,能调控细胞周期蛋白依赖性激酶1(cyclin-dependent kinase 1,CDK1)的磷酸化状态,从而调节CDK1与细胞周期蛋白B(cyclin B)复合物的活性从而实现对细胞周期的调控,且对DNA损伤检查点具有重要的调节作用。WEE1是G2/M期阻滞的关键基因,起着重要的监测作用,在一些癌症中过表达,抑制或下调WEE1激酶均能引发有丝分裂灾难,因此WEE1激酶抑制剂可能在抗癌治疗中有关键作用。在癌症的治疗过程中,WEE1抑制剂与DNA损伤剂、化学药物等联合使用会得到比单独使用更为有效,且在p53缺失的癌细胞中能发挥更好的效果。目前WEE1已成为许多癌症治疗的关键靶点之一,其抑制剂MK-1775已处于临床研究阶段,且能增强一些DNA损伤剂对p53缺失的癌细胞的杀伤能力。本文就WEE1激酶及其抑制剂在抗癌治疗中的应用作一综述。