Effects of phorbol esters on endothelial cell microfilaments: Laser scanning confocal microscopy and quantitative morphometry of dose dependent changes

Phorbol esters are known to alter microfilaments but it is not clear if the changes correspond to modulation of the phosphoinositide turnover/protein kinase C system. The novel technique of laser scanning confocal epifluorescence was used to study fiber orientation in phorbol ester treated cells. We treated endothelial cells with control agents and agents known to stimulate protein kinase C: 4 alpha-phorbol, phorbol 12-myristate 13-acetate (PMA), phorbol dibutyrate (PDB), or lipopolysaccharide. After incubation with the test agents, the endothelial cell microfilaments were stained with rhodamine pholloidin and viewed by conventional epifluorescence and by laser scanning confocal epifluorescence microscopy. The images obtained by the confocal microscopy corresponded to a thin optical section through the cells, 300 nm or more in thickness. The microfilaments extended predominantly in the plane of focus. After exposure of the cells to phorbol esters, the stress fibers became more nearly parallel in arrangement or were shortened, but remained in the plane of focus. The modification of microfilaments in response to phorbol esters was quantitated by a single blind analysis. In order to compare the morphological changes with a biochemical action of the phorbol esters, we measured phosphoinositide turnover. The dose-dependence of morphological changes was compared and contrasted to the dose-dependent effect of phorbol esters on bradykinin-stimulated phosphoinositide turnover. PMA had about the same EC50 (1-5 nM) for both biochemical and morphological processes. PDB was less potent in inducing the disruption of microfilament structure than in inhibiting phosphoinositide turnover. Lipopolysaccharide was ineffective in inducing a morphological change under these conditions. A simple activation of protein kinase C is insufficient to explain the dose-dependent effects of phorbol esters. Thus a morphometric analysis can help distinguish the potency of cytoskeleton modulators.     

Nuclear matrix involvement in sperm head structural organization

Summary— In the sperm nuclei the DNA is packaged into a highly condensed form and is not organized into nucleosome and solenoid but is bound and stabilized mainly by the protamines that arrange the DNA in an almost crystalline state. As demonstrated for somatic cells, the sperm DNA has been reported to be organized in loop domains attached to the nuclear matrix structures. However, the possible role of the sperm head matrix in maintaining the loop organization in absence of a typical nucleosomal structures has not been fully elucidated. By using in situ nick translation at confocal and electron microscope level, we analyzed the organization of the DNAprotamine complex and its association with the sperm nuclear matrix. The data obtained indicate that the chromatin organization in sperm nuclei is maintained during the sperm condensation by means of interactions with the nuclear matrix at fixed sites. The fine stucture of sperm nucleus and of sperm nuclear matrix, investigated on sections and replicas of freeze-fractured specimens, suggests that the lamellar array, observed by freeze-fracturing in the sperm nuclei, could depend on the inner matrix which presents a regular organization of globular structures possibly involved in the maintenance of chromatin domains in highly condensed sperm nuclei also.     

A dual fluorescence technique for visualization ofStaphylococcus epidermidis biofilm using scanning confocal laser microscopy

A new dual fluorescence technique is described which, when combined with scanning confocal laser microscopy (SCLM), can be used to visualize the components of biofilm produced byStaphylococcus epidermidis. Chemostat cultures of RP62A (a well-characterized slime-producing strain ofS. epidermidis) were used to produce mature biofilm on polyvinylcholoride (PVC) disks immobilized in a modified Robbins device using a seed and feed model system. Serial horizontal and vertical optical thin sections, as well as three-dimensional computer reconstructions, were obtained onin situ biofilm using the dual fluorescence procedure. Bacteria were visualized by green autofluorescence excited at 488 nm with an Argon laser. Cell-associated and exocellular matrix material (slime) was visualized by red fluorescence excited at 568 nm with a Krypton laser after interaction of the biofilm with Texas Red-labeled wheat germ agglutinin which is a slime-specific lectin marker. Structural analysis revealed that the cocci grew in slime-embedded cell clusters forming distinct conical-shaped microcolonies. Interspersed open channels served to connect the bulk liquid with the deepest layers of the mature, hydrated biofilm which increased overall surface area and likely facilitated the exchange of nutrients and waste products throughout the biofilm. The combined dual fluorescence technique and SCLM is potentially useful as a specific noninvasive tool for studying the effect of antimicrobial agents on the process of biofilm formation and for the characterization of the architecture ofS. epidermidis biofilm formedin vivo andin vitro on medical grade virgin or modified inert polymer surfaces.     

基于加速康复外科理念的早期康复训练联合八段锦前四式在乳腺癌改良根治术后患者中的应用效果研究

摘要 目的：研究基于加速康复外科（ERAS）理念的早期康复训练联合八段锦前四式在乳腺癌改良根治术后患者中的应用效果。方法：选择2019年9月~2022年5月期间在湖南中医药大学第一附属医院行乳腺癌改良根治术的128例乳腺癌患者。按照随机数字表法将患者分为对照组（基于ERAS理念的早期康复训练,n=64）和研究组（对照组的基础上接受八段锦前四式干预,n=64）。对比两组抑郁自评量表（SDS）评分、健康调查简表（SF-36）、焦虑自评量表（SAS）评分、Constant-Murley肩关节功能评分、疼痛视觉模拟评分（VAS）和肩关节活动度。结果：与对照组相比,研究组干预后SDS、SAS评分更低（P<0.05）。与对照组相比,研究组情感/生理职能、躯体疼痛、社会/生理功能、活力、总体/精神健康评分更高（P<0.05）。与对照组相比,研究组干预后VAS评分更低,Constant-Murley肩关节功能评分更高（P<0.05）。与对照组相比,研究组干预后前屈、后伸、外展关节活动度更大（P<0.05）。结论：基于ERAS的早期康复训练联合八段锦前四式应用于乳腺癌改良根治术后患者,可提高患者的生活质量,可能与缓解焦虑、抑郁情绪,减轻术后疼痛,促进肩关节功能恢复有关。     

不同剂量瑞芬太尼复合小剂量艾司氯胺酮用于腹腔镜妇科手术患者的效果及对麻醉效能、疼痛的影响

摘要 目的：研究不同剂量瑞芬太尼复合小剂量艾司氯胺酮对腹腔镜妇科手术患者麻醉效果的影响。方法：选取2020年4月～2023年2月在本院接受腹腔镜妇科手术治疗的60例患者进行研究,根据瑞芬太尼静脉输注剂量将其分为小剂量组、中剂量组和大剂量组,每组各20例。三组均给予患者小剂量艾司氯胺酮（0.1 μg?kg）,小剂量组给予患者0.1 μg/（Kg?min）瑞芬太尼,中剂量组给予患者0.3 μg/（Kg?min）瑞芬太尼,大剂量组给予患者0.5 μg/（Kg?min）瑞芬太尼。记录三组患者的麻醉起效时间、麻醉诱导时间、拔管时间和不良反应发生率,并检测其不同时间段的血流动力学和疼痛。结果：麻醉起效、麻醉诱导拔管时间比较,三组无显著差异（P＞0.05）。平均动脉压（MAP）和心率（HR）比较,大剂量组和中剂量组T1、T2时间段均低于小剂量组,中剂量组T1、T2时间段低于大剂量组;三组T2时间段MAP、HR水平低于T0、T1时间段,T1时间段低于T0时间段（P＜0.05）。疼痛视觉模拟量表（VAS）评分比较,大剂量组和中剂量组T1、T2时间段均低于小剂量组,中剂量组T1、T2时间段低于大剂量组;三组T2时间段MAP、HR水平低于T0、T1时间段,T1时间段低于T0时间段（P＜0.05）。不良反应发生率比较,三组无显著差异（P＞0.05）。结论：不同剂量瑞芬太尼复合小剂量艾司氯胺酮均能保障腹腔镜妇科手术的麻醉效果和安全性,但中剂量瑞芬太尼的应用更有利于稳定血流动力学,并减轻患者的疼痛程度。     

齿痛消炎灵颗粒联合牙周激光对牙周炎的应用效果及对口气和舌苔指标的影响

摘要 目的：探讨齿痛消炎灵颗粒联合牙周激光对牙周炎的应用效果及对口气和舌苔指标的影响。方法：选取我院2020年7月到2023年7月收治的80例牙周炎患者,分为观察组与对照组,各40例。对照组采取常规治疗与口服齿痛消炎灵颗粒,观察组以对照组为基础,增加牙周激光治疗。对比两组患者临床疗效,对比其治疗前与治疗4周后菌斑指数（PLI）,出血指数（BI）、探诊深度（PD）、临床附着丧失（CAL）相关牙周健康指标,口气感官值（OS）、口腔挥发硫化物（VSCs）相关口气指标以及舌苔厚度（Tt）、舌苔面积（Ta）相关舌苔指标变化。结果：观察组治疗总有效率较对照组高（P＜0.05）;治疗前两组患者CAL、PD、BI、PLI水平对无差异（P＞0.05）,治疗后两组均降低,且与对照组相比,观察组较低（P＜0.05）;治疗前两组患者OS、VSCs水平对比无差异（P＞0.05）,治疗后两组均降低,且与对照组相比,观察组较低（P＜0.05）;治疗前两组患者Tt、Ta评分对比无差异（P＞0.05）,治疗后两组均降低,且观察组较对照组低（P＜0.05）。结论：齿痛消炎灵颗粒联合牙周激光治疗牙周炎效果较好,改善患者牙周健康程度,减少舌苔情况,改善口气情况。     

一种目视激光显微镜

本文介绍一种目视激光显微镜。该装置采用白炽灯和激光做光源。通过调压器衬底亮度可以调整到零。由于激光的高亮度和强相干性,与普通显微镜相比,该显微镜具有景深长,分辨率高,层次丰富的特点。使用该显微镜时能实现镜象的假色彩编码,且镜象具有立体感。文中报导了该显微镜的原理和使用效果。     

Biophoton emission

The phenomenon of ultraweak photon emission from living systems was further investigated in order to elucidate the physical properties of this radiation and its possible source. We obtained evidence that the light has a high degree of coherence because of (1) its photon count statistics, (2) its spectral distribution, (3) its decay behavior after exposure to light illumination, and (4) its transparency through optically thick materials. Moroever, DNA is apparently at least an important source, since conformational changes induced with ethidium bromide in vivo are clearly reflected by changes of the photon emission of cells. The physical properties of the radiation are described, taking DNA as an exciplex laser system, where a stable state can be reached far from thermal equilibrium at threshold.