口腔黏膜白斑上皮组织中凋亡相关蛋白Bax的表达研究

目的探讨凋亡相关蛋白Bax在上皮增生性病变改变中的生物学行为及癌变与Bax蛋白的关系。方法采用免疫组织化学法检测法,观察分析29例口腔白斑、25例正常口腔黏膜上皮中细胞凋亡状况及凋亡相关蛋白Bax的表达。结果 Bax蛋白在白斑上皮组织中显过度表达。结论 Bax蛋白可能参与了增生性病变向恶性转化时的早期事件,起到一定的负调控作用。     

核因子-Bp65在口腔白斑及鳞癌中的表达

目的探讨核因子-Bp65在口腔白斑及鳞癌中的表达。方法应用免疫组织化学技术,检测正常口腔黏膜上皮(n=10),单纯增生型OLK(n=11),异常增生型OLK(n=35),OSCC(n=36)中的NF-Bp65表达情况。结果正常口腔黏膜上皮中p65不表达,由单纯增生型OLK转变到异常增生型OLK最后到OSCC的发生发展过程中p65的表达明显上调。结论p65可能是检测OLK的恶变潜能及OSCC早期诊断的敏感指标。     

坏死增生性淋巴结病细胞凋亡基因Bcl-2、Bcl-x及Bax的表达

目的 探讨坏死增生性淋巴结病,淋巴结细胞凋亡及Bcl-2、Bcl-x、Bax基因表达。方法颈部淋巴结活检,病理切片用光镜,电镜观察凋亡淋巴结细胞形态学变化,Western Blotting检测Bcl-2、Bcl-X、Bax基因蛋白表达变化。结果光镜、电镜可见坏死增生淋巴结病。有大量细胞凋亡,Westem Blotting检测结果显示Bcl-2、Bcl-x、Bax蛋白表达不同,与对照组比较,坏死增生性淋巴结病,抑凋亡基因Bcl-2蛋白表达水平明显下降（P〈0．05）,促凋亡基因Bax蛋白表达水平明显增加,Bcl—X蛋白表达改变不明显。结论坏死增生性淋巴结病可致调控凋亡基因的表达。导致细胞大量凋亡,淋巴结坏死。     

鼠急性全脑缺血再灌注后葛根素对海马CA1区Bcl-2、Bax表达的影响

目的　观察全脑缺血再灌注后葛根素对神经细胞凋亡相关基因Bcl 2、Bax表达的影响。方法　采用免疫组织化学法、原位末端标记法检测大鼠全脑缺血再灌注不同时间内海马CA1区Bcl 2和Bax蛋白表达水平及凋亡细胞数的变化。结果　①脑缺血再灌注后 ,海马CA1区Bcl 2蛋白的表达随再灌注时间不同而变化 ,缺血 10min后再灌注 6h达高峰 ;葛根素治疗组Bcl 2蛋白的表达于相应的时间点明显增加 ;②Bax蛋白表达在再灌注 2 4h达高峰 ,葛根素治疗组Bax蛋白表达在相应时间点则下降 ;③神经细胞凋亡数随再灌注时间延长而增加 ,葛根素可减少神经细胞凋亡数。结论　在全脑缺血再灌注后细胞凋亡中 ,Bcl 2、Bax发挥重要作用 ;葛根素通过上调Bcl 2蛋白的表达 ,下调Bax蛋白的表达抑制细胞凋亡发挥神经保护作用     

桥本甲状腺炎甲状腺组织Bcl-2、Bax蛋白表达

目的　探讨Bcl 2、Bax蛋白在桥本甲状腺炎 (HT)甲状腺组织的表达及其与HT发病机制的关系。方法　采用免疫组化S P法检测 4 1例HT及 10例正常甲状腺组织Bcl 2、Bax表达及分布 ,应用Mias99图像分析系统进行定量分析。结果　 (1)HT组甲状腺组织Bcl 2、Bax阳性颗粒面积、平均光密度和积分光密度均显著高于正常组 (P <0 0 1) ,HT组中组织结构破坏较重区域中小滤泡及失去正常滤泡结构的腺上皮Bax表达高于其它腺上皮 ,滤泡结构较完整的区域Bcl 2表达较高 ;(2 )HT甲状腺组织滤泡区Bcl 2阳性颗粒面积、积分光密度与Bax的比值低于正常对照组 (P <0 0 0 1)。结论　HT甲状腺组织Bcl 2、Bax表达失衡导致的细胞凋亡增高可能是HT甲状腺细胞破坏的重要机制之一 ;Bcl 2在维护HT甲状腺滤泡结构的完整性中起着重要作用     

脑缺血再灌注损伤后神经细胞凋亡及Bcl-2、Bax蛋白表达与罗非昔布的影响

目的观察大鼠局灶性脑缺血再灌注损伤后脑梗死面积的变化、脑组织神经细胞凋亡、Bcl2与Bax蛋白表达以及罗非昔布的保护作用。方法线栓法制备大鼠大脑中动脉闭塞2h/再灌注24h模型,于再灌注开始时灌胃给予罗非昔布。TTC染色观察脑梗死面积,TUNEL法检测神经细胞凋亡,免疫组化法检测脑组织Bcl2和Bax蛋白表达。结果损伤侧脑组织出现明显梗死灶,神经细胞凋亡率与Bax蛋白表达均明显升高,Bcl2/Bax比值明显下降。罗非昔布1.12、2.24mg·kg-1均可明显减少脑梗死面积,2.24mg·kg-1剂量还可显著降低神经细胞凋亡率与Bax蛋白表达,增高Bcl2蛋白表达与Bcl2/Bax比值。结论选择性COX2抑制剂罗非昔布可促进Bcl2蛋白表达并减少Bax蛋白表达,上调Bcl2/Bax比值而抑制神经细胞凋亡,从而明显改善缺血再灌注引起的脑损伤。     

三七总苷对人胃癌BGC-823细胞Bcl2和Bax表达的影响

目的:探讨三七总苷对胃癌BGC-823细胞Bcl2基因和Bax基因表达的影响。方法:不同浓度三七总苷诱导胃癌细胞BGC-823,实时荧光定量聚合酶链反应检测Bcl2基因和Bax基因的表达情况,免疫印迹检测Bcl2蛋白和Bax蛋白的表达情况,同时采用流式细胞仪检测细胞凋亡的改变。结果:三七总苷作用后,胃癌细胞Bcl2基因的表达受到明显抑制,而Bax基因的表达则明显上调,Bcl-2/Bax呈上升趋势;流式检测细胞增殖受到抑制。结论:三七总苷可促进胃癌BGC-823细胞凋亡,具有抗胃癌细胞恶性增殖活性。     

4种归肝经明目中药对晶状体上皮细胞凋亡相关基因Bcl-2和Bax的调控

目的 :探讨 4种归肝经明目中药车前子、青葙子、菊花和熟地对实验性氧化损伤大鼠晶状体上皮细胞 (lensepithelialcell,LEC)凋亡相关基因Bcl 2和Bax的调控。方法 :将SD大鼠双眼随机分成 7组 :空白对照组、过氧化氢 (H2 O2 )组、吡诺克辛 (PS)组、车前子组、青葙子组、菊花组和熟地组。无菌操作摘除眼球并在手术显微镜下分离晶状体 ,使晶状体孵育在 30 0 μmol·L-1H2 O2 培养液中复制LEC凋亡模型 ,同时采用 4种归肝经明目中药干预 ,置二氧化碳培养箱共同孵育 2 4h。取晶状体前囊膜采用免疫组化法检测凋亡相关基因Bcl 2和Bax的蛋白表达并进行比较。结果 :正常SD大鼠LEC中Bcl 2和Bax均有表达 ,Bcl 2表达较Bax表达强 ;H2 O2组Bcl 2表达显著下调 ,Bax表达显著上调 (P <0 .0 1) ,Bcl 2 Bax比率下降 ;与H2 O2 组比较 ,4种归肝经明目中药可明显上调Bcl 2表达 ,下调Bax表达(P <0 .0 1) ,Bcl 2 Bax比率上升 ;PS与 4种归肝经明目中药的调节作用相似 ,但弱于 4种归肝经明目中药的作用 (P <0 .0 5 )。结论 :4种归肝经明目中药调控凋亡相关基因Bcl 2和Bax的表达可能是其抑制LEC凋亡的分子机制。     

Bcl－2与卵巢颗粒细胞凋亡相关性研究

目的：比较卵巢功能减退患者和卵巢功能正常女性颗粒细胞凋亡及 Bcl－2表达情况,探讨卵巢功能减退的发病机制。方法选择接受体外受精－胚胎移植（IVF－ET）治疗的不孕患者48例,其中卵巢功能减退患者23例,卵巢功能正常对照患者25例。于取卵日收集卵泡液,离心收集颗粒细胞培养,用 TUNEL 检测颗粒细胞凋亡情况。免疫组化分析颗粒细胞 Bcl－2蛋白的表达情况。结果卵巢功能减退组和卵巢功能正常组颗粒细胞凋亡率分别为（27±13）％、（18±8）％,卵巢功能减退组颗粒细胞凋亡率较对照组增高（ P ＜0．05）;免疫组化分析显示 Bcl－2主要在颗粒细胞胞浆中表达,卵巢功能减退组 Bcl－2蛋白表达较正常对照组下调（ P ＜0．05）。结论 Bcl－2可能通过调节颗粒细胞凋亡情况,影响卵泡发育及卵巢的功能。     

澳洲茄边碱对肝癌 HepG2细胞增殖及凋亡的影响

目的：探讨澳洲茄边碱（SM ）对 HepG2细胞增殖、凋亡的影响及其可能作用机制。方法用不同浓度SM 5、10、15和20μg／ml分别处理 HepG2细胞3、6、12和24 h ,并设不加药的对照组。采用MTT法检测 HepG2细胞增殖,DAPI染色法观察细胞核形态的变化,流式细胞术检测细胞凋亡和周期,Western blot法检测B细胞淋巴瘤‐白血病2（Bcl‐2）、Bcl相关X蛋白（Bax）、半胱天冬氨酸蛋白酶3（Caspase‐3）及Ki67的蛋白表达。结果与对照组相比,SM 呈剂量依赖性地抑制HepG2细胞增殖,促进HepG2细胞凋亡,将细胞周期阻滞于G2／M期,并且上调Bax和Caspase‐3蛋白表达,下调Bcl‐2和Ki67蛋白表达（P＜0．05或P＜0．01）。结论 SM 能有效抑制 HepG2细胞的增殖,促进细胞凋亡的发生;SM上调Bax和Caspase‐3表达,下调Bcl‐2和Ki67表达,细胞周期阻滞于G2／M期可能是其发挥上述作用的机制。     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.