不同浓度过氧乙酸灭活HBV效果观察

本文通过对0．2％0．3％0．4％……1．1％十种不同浓度的过氧乙酸作用不同时间，观察其对HBV灭活效果，发现有三点与文献报告有所不同：①HBsAg抗原性的灭活时间延长：0．2％180分钟、0．3％120分钟、0．4％60分钟、0．5％60分钟、0．6％30分钟，浓度在0．7％以上者5分钟内即可灭活。②HBV—DNA灭活所需过氧乙酸浓度降低及时间缩短，0．2％5分钟不能灭活HBV—DNA，超过5分钟或浓度大于0．2％者5分钟内即可灭活HBV—DNA。③HBsAg抗原性的消失与HBV—DNA活性消失存在一定距离对BSAg抵抗化学因子较HBV—DNA强，破坏HBsAg抗原性较HBV－DNA难，提示在观察和检测消毒剂灭活HBV效果时应同时观察HBsAg、HBV－DNA，在实际操作中，应加大消毒剂浓度或延长消毒时间，以快速彻底地杀灭HBV。     

研制乙型肝炎疫苗的实验:乙型肝炎表面抗原对豚鼠的免疫原性

用凝胶过滤、等比带离心及区带离心法,从人血浆提纯乙型肝炎表面抗原(HBsAg)疫苗。最后制品的活性为原有活性的60％,基本上不含乙型肝炎病毒(HBV)和血浆蛋白。所提纯的HBsAg经0.1％浓度的甲醛处理后,已使残余感染性灭活,但其体外试验的抗原性及对豚鼠的免疫原性均未有显著降低。     

3种环糊精对亚甲蓝增敏作用的考察

目的:考察3种常见环糊精(α-环糊精、β-环糊精、γ-环糊精)对亚甲蓝增敏的作用。方法:以亚甲蓝水溶液为基础,以及亚甲蓝的特征荧光强度为标准,采用荧光分光光度法考察3种环糊精在不同浓度时α-环糊精、β-环糊精和γ-环糊精对亚甲蓝的荧光增敏作用的强度。结果:β-环糊精对亚甲蓝有增敏作用,而α-环糊精、γ-环糊精无增敏作用。结论:选择β-环糊精的荧光增敏强度作为亚甲蓝残留量的检测,用于提高检测亚甲蓝残留量的检测限、灵敏度及准确度。     

大肠杆菌表达的乙型肝炎表面抗原124aa分子的纯化和免疫学鉴定

目的探索大肠杆菌表达的乙型肝炎表面抗原(HBsAg)124aa分子作为疫苗成分应用的可能性. 方法以超声破碎法获取包涵体,用HisTrapTM试剂盒亲和层析纯化目的蛋白,加入聚乙二醇(PEG4000)以提高变性蛋白的复性效率;以抗HBsAg a抗原决定簇单克隆抗体(McAb)和抗HBsAg多克隆抗体(PcAb)作为包被抗体,采用夹心 ELI SA法分析其抗原性;以纯化产物免疫BALB/c小鼠,RIA法测定小鼠血清中抗HB s抗体.结果通过HisTrapTM试剂盒亲和层析纯化的HBsAg124aa分子经反相高效液相色谱(RP-HPLC)分析,纯度达95 %以上 ;经复性后的蛋白具有抗原性和免疫原性.结论 HBsAg124aa 分子在大肠杆菌中的成功表达,为探索新的乙型肝炎疫苗成分提供了重要线索.     

“复方亚甲蓝”在肢体坏死的局部镇痛治疗

目的：总结“复方亚甲蓝”在肢体坏死的局部镇痛治疗方法和效果。方法：以“复方亚甲蓝”对32例剧烈疼痛肢体坏死创面行局部湿敷，评价镇痛效果。结果：32例患者经治疗后均有不同程度的镇痛作用。结论：“复方亚甲蓝”在肢体坏死的局部镇痛治疗方面具有经济方便的优点，可以减少毒麻类药品的应用，镇痛效果显著，故不失为一种好的方法，值得推广应用。     

211种贵州民族中草药抑制HBsAg的体外实验研究

目的筛选贵州民族中草药对HBsAg的作用。方法选取211种贵州省民族中草药提取物分别与HBsAg37℃作用1小时,通过ELISA双抗体夹心法分别检测HBsAg与药物作用前后的含量,计算HBsAg经药物作用后下降的百分率,筛选出有效的药物。结果211种药物中筛选出24种药物有抗HBsAg作用,占总数的11．4％。结论贵州民族中草药中有部分药物对HBsAg的抑制作用。     

大肠杆菌表达的乙型肝炎表达抗原124aa分子的纯化和免疫学鉴定

目的 探索大肠杆菌表达的乙型肝炎表面抗原（HBsAg）124aa分子作为疫苗成分应用的可能性。方法 以超声破碎法获取包涵体,用HisTrap^TM试剂盒亲和层析纯化目的蛋白。加入聚乙二醇（PEG4000）以提高变性蛋白的复性效率。以抗HBsAga抗原决定簇单克隆抗体（McAb)和抗HBsAg多克隆抗体（PcAb)作为包被抗体,采用夹心ELISA法分析其抗原性;以纯化产物免疫BALB/c小鼠,RIA法测定小鼠血清中抗HBs抗体。结果 通过HisTrap^TM试剂盒亲和层析纯化的HBsAg124aa分子经反相高效液相色谱（RP-HPLC）分析。纯度达95%以上,经复性后的蛋白具有抗原性和免疫原性。结论 HBsAg124aa分子在大肠杆菌中的成功表达,为探索新的乙型肝炎疫苗成分提供了重要线索。     

肝炎病毒消毒剂应用现状

长期以来,对病毒性肝炎的防治,采取以切断传播途径为主导的综合措施。为此科学家广泛研究各种新的物理或化学消毒方法,以筛选出更多价廉、高效的化学消毒剂为目的,满足临床实际的需要。一、消毒剂的考核指标甲型肝炎病毒(HAV)已能组织培养,可以用组织培养及动物感染的方法来直接测定理化因子来考核消毒剂作用后对HAV灭活情况。乙型肝炎病毒(HBV)迄今不能培养,故消毒指标的研究仍然是一个值得探讨的课题。目前常以HBsAg抗原性消失作为HBV传染性消失的指标。消毒剂对HBsAg的型态破坏可用电镜观察等方法,如     

氧化电位水消毒剂性能与杀菌效果实验研究

为了解氧化电位水消毒剂的性能及消毒效果，用加速法作稳定性试验，用称量法作金属腐蚀性试验，用悬液法作微生物杀灭效果试验，结果表明，在54℃下放置2周，A剂有效浓度下降43．66％，对不锈钢无腐蚀，对碳钢，铝和铜均为中度腐蚀；腐蚀速率R分别为：0．000、0．2374、0．2551和0.2565mm/a。100％杀灭大肠杆菌（8099）、金黄色葡萄球菌（ATCC6538）、白色念珠菌（ATCC10231）、枯草杆菌黑色变种芽胞（ATCC9372）的剂量分别为：45mg/L作用10min,64mg/L作用15min,100mg/L作用25min,250mg/L作用20min，HBsAg的抗原性被破坏所需剂量为240mg/L作用2min。结论：氧化电位水消毒剂稳定性好，对不锈钢无腐蚀，对碳钢，铝和铜中度腐蚀，对细菌杀灭作用强，对HBsAg破坏作用迅速。     

亚甲蓝长效镇痛剂对肛肠病术后镇痛的观察

目的:观察不同浓度复方亚甲蓝制剂在肛肠病术后镇痛的疗效及安全性.方法:将427例患者随机分为高浓度组和低浓度组,其中高浓度组使用亚甲蓝浓度为0.3%,低浓度组使用亚甲蓝浓度为0.08%,观察2组止痛效果及不良反应.结果:高浓度组与低浓度组在止痛效果方面无明显差异(P＞0.05),而在术后和近期不良反应方面和远期不良反应方面,低浓度组比高浓度组低(P＜0.05).结论:综合考虑疗效及不良反应情况,采用复方亚甲蓝制剂作长效镇痛低浓度优于高浓度.     

亚甲蓝对结肠癌细胞系Lovo的杀伤和抑制作用

为探讨亚甲蓝对体外培养的人大肠癌细胞的抗肿瘤作用 ,采用直接杀伤实验、细胞生长抑制实验和细胞集落形成能力实验方法研究了亚甲蓝对体外培养的人大肠癌细胞系Lovo的作用 ,同时用透射电镜技术观察了该药对癌细胞的损伤部位 .结果显示 13 2 5 μmol/L亚甲蓝作用2 4h可杀死 79 4%的大肠癌细胞 ,5 3μmol/L亚甲蓝作用 2 4h可杀死全部大肠癌细胞 ,0 2 7μmol/L亚甲蓝即可明显抑制人结肠癌细胞的增殖 .透射电镜观察显示亚甲蓝损伤细胞的部位在线粒体 .研究结果表明 ,亚甲蓝对体外培养的人大肠癌细胞Lovo具有显著的杀伤和抑制作用 ,其损伤癌细胞的部位在线粒体     

基于阴离子型环糊精超分子包合物高灵敏荧光光度分光法检测血浆中残留亚甲蓝

目的:建立检测血浆中微量亚甲蓝的高灵敏荧光分光光度法。方法:利用羧甲基-β-环糊精对亚甲蓝荧光信号的增敏作用,用荧光分光光度法直接测定血浆中的微量亚甲蓝。结果:亚甲蓝检测浓度在0.008⁰.2μmol/L范围内线性关系良好(r=0.999 0)。包合反应20 min后体系荧光强度稳定,20、40、60、120 min的RSD为0.57%。高、中、低浓度的日内RSD分别为0.36%、0.53%和0.92%,日间RSD分别为2.31%、2.62%和5.43%;回收率为97.04%0.2μmol/L范围内线性关系良好(r=0.999 0)。包合反应20 min后体系荧光强度稳定,20、40、60、120 min的RSD为0.57%。高、中、低浓度的日内RSD分别为0.36%、0.53%和0.92%,日间RSD分别为2.31%、2.62%和5.43%;回收率为97.04%¹07.70%。20个实际样品亚甲蓝残留量均在线性范围内,且低于规定最大残留量。结论:本方法准确、简便、快速、灵敏度高,可直接用于血浆中微量亚甲蓝的测定。     

亚甲蓝类似物的合成及其与牛血清白蛋白相互作用的研究

目的 合成3种亚甲蓝类似物3,7-二(二正丙胺基)-吩噻嗪-5-鎓碘化物、3,7-二(二正丁胺基)-吩噻嗪-5-鎓碘化物和3,7-二(二正戊胺基)-吩噻嗪-5-鎓碘化物,并研究模拟生理条件下这3种亚甲蓝类似物与牛血清白蛋白的相互作用.方法 通过1H-NMR及MS对这3种亚甲蓝类似物进行结构表征,应用荧光光谱法研究了模拟生理条件下这3种亚甲蓝类似物与牛血清白蛋白的相互作用.结果 3种亚甲蓝类似物和BSA相互作用形成了蛋白质-药物复合物并猝灭其固有荧光,亚甲蓝类似物与BSA相互作用过程的ΔG0<0,亚甲蓝类似物与BSA荧光残基间的距离r均小于7 nm.结论 亚甲蓝类似物和BSA相互作用的猝灭机制为静态猝灭,二者之间的反应是自发进行的.非辐射能量转移理论表明BSA与亚甲蓝类似物之间存在非辐射能量转移.同步荧光光谱和三维荧光光谱的研究结果表明,亚甲蓝类似物与BSA的相互作用导致BSA构象发生变化.     

亚甲基蓝对脑缺血再灌注大鼠血脑屏障的保护作用

目的 探讨亚甲基蓝对大鼠脑缺血再灌注损伤的保护作用及机制.方法 SD大鼠随机分为假手术组、模型组、亚甲基蓝低剂量组(1 mg/kg)、亚甲基蓝中剂量组(2 mg/kg)、亚甲基蓝高剂量组(4 mg/kg),每组12只.线栓法制备大鼠左侧颈动脉栓塞2 h再灌注24 h模型.亚甲基蓝组于术前1 d,再灌注时腹腔注射相应剂量...     

Cation exchange resins as pharmaceutical carriers for methylene blue: Binding and release

Methylene blue is a competitive inhibitor of the glutathione reductase of Plasmodium falciparum and is used in combination with other antimalarial drugs leading to a renaissance of methylene blue in malaria therapy. Its bitter flavour and tissue colouring property impair compliance, especially in children. These problems may be solved by binding the cationic methylene blue to cation exchange materials as pharmaceutical carriers in order to mask the undesirable properties. However, such carriers are only useful if the antimalarial is released under physiological conditions. The binding to seven cation exchange resins was studied. Ion exchangers on acrylic or methacrylic acid basis bound between 1.54 and 2.16 g methylene blue chloride trihydrate per gram ion exchanger. Polymers on divinylbenzene or styrene basis with sulphonic acid groups bound 306 and 384 mg of methylene blue chloride trihydrate per gram ion exchanger. In aqueous solution at pH of 1.5, nearly all bound methylene blue was released. The release of methylene blue from (meth)acrylic acid polymers in the presence of proteins and fat was not affected. From these data ion exchangers present a promising group of pharmaceutical carrier for the safe and compliant drug administration of methylene blue to children.     

伊立替康联合亚甲蓝示踪进展期胃癌淋巴结的研究

目的:探讨伊立替康联合亚甲蓝在进展期胃癌根治术中淋巴结示踪的效率及其机制。方法:90例进展期胃癌随机分为非示踪组(31例)、亚甲蓝示踪组(30例)和伊立替康联合亚甲蓝示踪组(29例,简称联合示踪组),其中亚甲蓝示踪组术中用亚甲蓝2 mL分4点于瘤周浆膜下注射,联合示踪组用40 mg伊立替康与2 mL亚甲蓝耦合后分4点注射。3组均用多功能手术解剖器进行刮吸法胃癌根治淋巴结清扫。分别统计各组手术时间、出血量、输血量、清除淋巴结数和术后随访情况。结果:非示踪组、亚甲蓝示踪组和联合示踪组平均手术时间分别为(218±67)、(192±31)和(205±36)min(P>0.05);平均出血量分别为(248±116)、(164±88)和(173±98)mL(P<0.05),输血者平均输血量分别为(457±159)(、489±176)和(467±148)mL(P>0.05);平均每例清除淋巴结数分别为(22±9)、(22±10)和(30±9)枚(P<0.05),平均每例阳性淋巴结数分别为(7±3)、(6±3)和(9±3)枚(P<0.05);3组分别有29、28和28例得到平均3年的随访,肿瘤复发率分别为31.0%(9/29)、32.1%(9/28)和21.4%(6/28)(P>0.05),3年存活率分别为62.1%(18/29)、57.1%(16/28)和78.6%(22/28)(P>0.05)。结论:瘤周浆膜下注射伊立替康与亚甲蓝耦合液可明显提高进展期胃癌患者淋巴结清除效率,且安全简便。其机制可能是耦合液显著延长淋巴结染色和褪色时间,始终存在色觉导向刺激,激发术者进行准确而彻底的淋巴结清扫。     

Pharmacokinetics and organ distribution of intravenous and oral methylene blue

OBJECTIVE: To determine the pharmacokinetics and organ distribution of i.v. and oral methylene blue, which is used to prevent ifosfamide-induced encephalopathy in oncology. METHODS: The concentration of methylene blue in whole blood was measured using high-performance liquid chromatography in seven volunteers after i.v. and oral administration of 100 mg methylene blue with and without mesna. The distribution of methylene blue in different tissues was measured in rats after intraduodenal and i.v. application. RESULTS: The time course of methylene blue in whole blood after i.v. administration showed a multiphasic time course with an estimated terminal half-life of 5.25 h. Following oral administration, the area under the concentration-time curve was much lower (9 nmol/min/ml vs 137 nmol/min/ml). Co-administration of mesna, which could influence distribution by ion-pairing, did not alter the pharmacokinetics. The urinary excretion of methylene blue and its leucoform was only moderately higher after i.v. administration (18% vs 28% dose). Intraduodenal administration to rats resulted in higher concentrations in intestinal wall and liver but lower concentrations in whole blood and brain than i.v. methylene blue. CONCLUSIONS: Differences in organ distribution of methylene blue are mainly responsible for the different pharmacokinetics after oral and i.v. administration. If methylene blue acts in the liver, where ifosfamide is primarily activated to reactive and potentially toxic metabolites, oral and i.v. methylene blue are likely to be equally effective. However, if the site of action is the central nervous system, i.v. methylene blue which results in much higher concentrations in brain seems preferable.     

增敏荧光法测定血浆中亚甲蓝含量及其残留量检测

目的 建立检测血浆中微量亚甲蓝的增敏荧光分光光度法.方法 利用β-环糊精对亚甲蓝荧光信号的增敏作用,以665nm为激发波长,680nm为发射波长,用荧光分光光度法直接测定血浆中的微量亚甲蓝.结果 用β-环糊精增敏时,亚甲蓝浓度在0.012~2.283μmol/L范围内呈现良好的线性关系（r=0.9994）,高中低浓度的日内变异系数（CV）分别为0.19%、0.41%和 1.14%,日间CV分别为1.76%、2.24%和2.97%,标准曲线法测得回收率为99.39~102.30%,对照品比较法测得回收率为96.93%~104.02%.结论 本研究体现出了简单,准确、快捷的特点,实现了无需进行分离、富集达到直接测定血浆中亚甲蓝浓度的目的.     

Comparative studies on the effects of toluidine blue and methylene blue on the reduction of ferrihaemoglobin in man and dog

Summary In humans and in dogs, respectively, about 30 and 60% of the haemoglobin was oxidized to ferrihaemoglobin by intravenous injection of 4-dimethylaminophenol hydrochloride. Toluidine blue was found to accelerate the subsequent reduction of ferrihaemoglobin in dogs 2–3 times more rapidly than methylene blue. In man toluidine blue 2 mg/kg was about twice as effective as the same dose of methylene blue. Such doses of methylene blue caused side effects, and according to the dose-effect relationship a further increase in the rate of ferrihaemoglobin reduction with a further increase in the dose of catalyst was unlikely. Toluidine blue 4 mg/kg did not produce the side effects observed with methylene blue and accelerated the reduction of ferrihaemoglobin by 60% more than doses of 2 mg/kg. Its greater maximal effect, and the fact that it is better tolerated than methylene blue, imply that toluidine blue should be preferred for accelerating ferrihaemoglobin reduction in man.     

Erythrocyte membrane alterations as the basis of chlorate toxicity

The effects of sodium chlorate and of sodium nitrite on human erythrocytes were studied in vitro. Nitrite rapidly oxidised haemoglobin and glutathione; reduction of methaemoglobin (Hbi) by methylene blue was complete during 3 h of incubation with nitrite. With chlorate, a concentration-dependent lag phase was seen before Hbi was formed. After prolonged incubation, Hbi could no longer be reduced with methylene blue. Several other effects were observed that explain the clinical picture of chlorate poisoning which involves haemolysis followed by disseminated intravascular coagulation and renal failure: increased permeability to cations, increased resistance to hypotonic haemolysis and prolonged filtration time through polycarbonate membranes with cylindrical pores of 5 micron diameter. This suggests an increased membrane rigidity due to membrane protein polymerisation, as demonstrated by SDS polyacrylamide gel electrophoresis. Simultaneously, erythrocyte enzymes were inactivated, primarily glucose-6-phosphate dehydrogenase which is necessary for the therapeutic effect of methylene blue. This explains the inefficacy of methylene blue in the treatment of a case of chlorate poisoning that we observed (Arch. Toxicol., 48 (1981) 281).