Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity

In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77 EU/mg and a yield of 35.5% using Sepharose-4B-l-tyrosine- sulphanilamide affinity column chromatography. For determining the enzyme purity and subunit molecular mass, sodiumdodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The molecular mass of subunit was found approximately 29.71 kDa. The optimum temperature, activation energy (E_a), activation enthalpy (ΔH) and Q₁₀ values were obtained from Arrhenius plot. K_m and V_max values for p-nitrophenyl acetate of the purified enzyme were calculated from Lineweaver-Burk graphs. In addition, the inhibitory effects of different heavy metal ions (Fe²⁺, Pb²⁺, Co²⁺, Ag⁺ and Cu²⁺) on Black Sea trout kidney tissue CA enzyme activities were investigated by using esterase method under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC₅₀) were obtained. Finally K_i values and inhibition types were calculated from Lineweaver-Burk graphs.     

乌司他丁与连续性血液净化联合治疗热射病的效果及对机体炎性反应影响研究

目的 分析乌司他丁与连续性血液净化联用对热射病患者所产生的影响。方法 选取2017年5月—2019年2月在海南医学院第二附属医院进行治疗的88例热射病患者作为研究对象,将其分为研究组和常规组,常规组患者接受常规治疗,研究组患者在常规治疗基础上,将乌司他丁与连续性血液净化联用。分析两种治疗措施的效果。结果 治疗前两组患者的IL-17、TNF-α、cTnⅠ、β₂-MG、PT、D-D、ET和TM水平,差异无统计学意义（P>0.05）。经治疗两组患者的上述指标水平都表现出下降的趋势,相对于常规组,研究组患者下降水平更明显,差异具有统计学意义（P<0.05）。研究组治疗的总有效率为95.5%,高于常规组的72.7%,差异具有统计学意义（P<0.05）。结论 运用乌司他丁与连续性血液净化联用的疗法对热射病患者进行治疗,可获得十分理想的治疗效果,可在临床工作中推广。     

The Relationship Between Air Pollution and Lung Cancer in Nonsmokers in Taiwan

Introduction

For never-smokers (smoked <100 lifetime cigarettes), lung cancer (LC) has emerged as an important issue. We aimed to investigate the effects of prevalence changes in tobacco smoking and particulate matter (PM) 2.5 (PM_2.5) levels on LC in Taiwan, in relation to contrasting PM_2.5 levels, between Northern Taiwan (NT) and Southern Taiwan (ST).

Extracorporeal blood purification treatment options for COVID-19: The role of immunoadsorption

The activation of the innate and adaptive immune systems by SARS-CoV-2 causes the release of several inflammatory cytokines, including IL-6. The inflammatory hypercytokinemia causes immunopathological changes in the lungs including vascular leakage, and alveolar edema. As a result of these changes in the lungs, hypoxia and acute respiratory distress syndrome occur in patients with COVID-19. Even though there are clinical trials on the development of therapeutics and vaccines, there are currently no licensed vaccines or therapeutics for COVID-19. Pharmacological approaches have shown poor results in sepsis-like syndromes caused by the hypercytokinemia. Suppressing the cytokine storm is an important way to prevent the organ damage in patients with COVID-19. Extracorporeal blood purification could be proposed as an adjunctive therapy for sepsis, aiming to control the associated dysregulation of the immune system, which is known to protect organ functions. Several extracorporeal blood purification therapies are now available, and most of them target endotoxins and/or the cytokines and aim improving the immune response. For this purpose, plasmapheresis and immunoadsorption may be an important adjunctive treatment option to manage the complications caused by cytokine storm in critically ill patients with COVID-19.     

生物安全柜的控制

生物安全柜是重要的一级屏障设备，适当的控制方式对生物安全柜的防护能力是很重要的。文中主要介绍了生物安全柜的控制要点和几种控制方式及其特点。     

从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位

目的：建立一种有效从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位HspA的方法。方法：重组基因工程大肠杆菌发酵后，表达的Hp r HspA包涵体经洗涤，变性，复性，采用Q Sepharose High Performance阴离子交换层析和Superdex75凝胶过滤层析分离纯化，使用SDS－PAGE和HPLC检测纯度，选用ELISA和动物实验对纯化蛋白的免疫学活性和生物学活性进行鉴定。结果：Hp的HspA包涵经洗涤和溶解后，Hp的HspA的纯度＞60％，包涵体溶解液经阴离子交换层析和凝胶过滤层析后纯度超过95％，Hp的HspA纯品经检测具有良好的免疫学活性和生物学活性。结论：本研究建立的分离纯化方法可从包涵体中获得高纯度的重组HspA蛋白，为进一步的动物实验研究奠定了基础。     

港九海底铁路建设中的减压病——工作条件与发病关系探讨

本文介绍香港九龙海底铁路建设中高气压作业与减压病发病情况。该工程有11条坑道,共计259 448个高压工作日,减压288 140人次,发生减压病1 534例。根据不同的工作压力,以暴露次数、高压工作日数及暴露4小时以上次数3种方法分别计算减压病发病率。结果表明:各种方法计算出的发病率都有随工作压力增高而上升的趋势,说明现用的减压表对不同的工作压力达不到同样的预防效果。本文还对3大主要工种和14个小工种的减压病作了分析。     

胞嘧啶脱氨酶的原核表达和多克隆抗体的制备

目的：在原核系统中表达并纯化大肠杆菌胞嘧啶脱氨酶（cytosine deaminase,CD)，制备鼠抗大肠杆菌CD多克隆抗体，方法：亚克隆CD基因到原核表达载体pMAL-c2和pBV222中，并转化入大肠杆菌DH5α内，诱导表达并纯化MBP-CD和6his-CD融合蛋白，用纯化的MBP-CD融合蛋白免疫小鼠制备多克隆抗体。结果：通过重组质粒的酶切筛选出重组阳性克隆，成功地表达和纯化出MBP-CD和6his-CD融合蛋白，用纯化的MBP-CD成功制备了鼠抗CD多克隆抗体，并用6his-CD和GST-CD重组蛋白进行Western印迹分析，证实了抗体的正确性，结论：应用多克隆抗体可以检测体内外CD基因的表达，为临床前和临床上深入开展CD基因的生物治疗研究提供重要的实验材料。