羟胺三氯化铁法测定人血清胆碱酯酶方法的优化

目的： 鉴于羟胺三氯化铁法（WS/T66-1996）测定人血清胆碱酯酶（ChEs）时效率较低以及容易产生沉淀等缺陷，本研究旨在对该方法进行优化，以使测定更加高效。方法： 首先验证羟胺三氯化铁法测定AChE及BChE的适用性。在此基础上，进一步研究微孔板羟胺三氯化铁法测定BChE的适用性。最后，以改良Ellman法为参考，对优化前后的准确性及可靠性进行验证。结果： 羟胺三氯化铁法测定人血清中AChE酶活性的灵敏度较差，但是对人血清中BChE酶活性的测定结果较为理想。羟胺三氯化铁法与改良Ellman法测定BChE酶活性测定结果之间呈正相关关系（r=0.652，P < 0.01）；微孔板羟胺三氯化铁法与羟胺三氯化铁法测定BChE酶活性测定结果之间也呈正相关关系（r_s=0.653，P < 0.01）。与羟胺三氯化铁法方法相比，微孔板羟胺三氯化铁法的准确性及可靠性略佳，主要体现在最低检测浓度及变异系数上。结论： 微孔板羟胺三氯化铁法是一种通量较高、经济性较好的测定人血清BChE酶活性的方法，有一定的推广应用前景。     

协同刺激分子B7-H4在小鼠食管癌前病变中的表达

目的：研究协同刺激分子B7同系物4（B7-H4）在小鼠食管癌前病变中的表达变化,探讨其在食管鳞状细胞癌发生中的作用。方法：133只C57BL/6小鼠,分为对照组42只和诱癌组91只。诱癌组小鼠采用饮水法给予50 μg/mL 4-硝基喹啉-1-氧化物（4NQO）15周,诱导小鼠食管癌前病变。通过苏木素-伊红（HE）染色法评价食管组织病理变化;免疫组化和Western blot检测B7-H4蛋白在食管组织中的表达,Western blot检测B7-H4蛋白在小鼠血清中的表达。结果：从16~28周,4NQO诱癌组小鼠食管组织出现肉眼可见的增厚、凸起、肿块等病理改变。HE染色结果显示,小鼠食管组织癌前病变级别与诱癌时间呈正相关（r=0.55,P < 0.01）。另外,B7-H4在食管组织的表达水平与食管癌前病变级别呈正相关（r=0.57,P < 0.01）。与对照组相比,第26周和第28周诱癌组小鼠食管组织B7-H4蛋白表达显著升高（P < 0.05或P < 0.01）。与正常小鼠相比,高级别上皮内瘤变小鼠血清B7-H4蛋白浓度显著升高（P < 0.05）。结论：小鼠食管癌前病变进程中,B7-H4蛋白在血清和食管组织中表达上调,B7-H4可能参与了食管癌前病变的进程。     

基于生物信息学数据库探讨FOXO3基因在膀胱癌中的表达及临床意义

目的：探讨叉头转录蛋白O亚族3（FOXO3）基因在膀胱癌中的表达及临床意义。方法：利用GEPIA数据库分析FOXO3基因在膀胱癌与其癌旁正常组织中的表达情况,及其与患者预后的关系,利用UALCAN数据库分析膀胱癌中FOXO3表达与淋巴结转移的关系;通过TIMER数据库分析FOXO3基因表达与膀胱癌免疫浸润水平的相关性;通过String数据库分析与FOXO3相互作用的蛋白。结果：GEPIA数据库分析结果显示,FOXO3基因在膀胱癌组织中的表达显著低于癌旁正常组织（P < 0.01）;低表达FOXO3基因的患者总生存率（OS）和无疾病生存率（DFS）均显著高于高表达者（P < 0.05）。UALCAN数据库分析结果显示膀胱癌中FOXO3的表达与淋巴结转移无明显相关。TIMER数据库分析显示FOXO3基因表达与B细胞（r=0.133,P < 0.05）、CD8⁺T细胞（r=0.262,P < 0.01）、CD4⁺T细胞（r=0.12,P < 0.05）、巨噬细胞（r=0.252,P < 0.01）、中性粒细胞（r=0.242,P < 0.01）和树突状细胞（r=0.162,P < 0.01）的免疫浸润水平呈正相关。蛋白相互作用网络分析发现,AKT1、SIRT1、EP300、BCL2L11、SOD2、SGK1、AKT2、CREBBP、CDKN1B、SMAD4等蛋白与FOXO3具有明显相互作用。结论：基于肿瘤基因数据库分析发现,FOXO3基因在膀胱癌组织中低表达,其高表达与患者预后不良相关;FOXO3表达与膀胱癌免疫细胞的浸润水平有关。     

基于生物信息学数据库探讨FOXO3基因在膀胱癌中的表达及临床意义

目的：探讨叉头转录蛋白O亚族3（FOXO3）基因在膀胱癌中的表达及临床意义。方法：利用GEPIA数据库分析FOXO3基因在膀胱癌与其癌旁正常组织中的表达情况，及其与患者预后的关系，利用UALCAN数据库分析膀胱癌中FOXO3表达与淋巴结转移的关系；通过TIMER数据库分析FOXO3基因表达与膀胱癌免疫浸润水平的相关性；通过String数据库分析与FOXO3相互作用的蛋白。结果：GEPIA数据库分析结果显示，FOXO3基因在膀胱癌组织中的表达显著低于癌旁正常组织（P < 0.01）；低表达FOXO3基因的患者总生存率（OS）和无疾病生存率（DFS）均显著高于高表达者（P < 0.05）。UALCAN数据库分析结果显示膀胱癌中FOXO3的表达与淋巴结转移无明显相关。TIMER数据库分析显示FOXO3基因表达与B细胞（r=0.133，P < 0.05）、CD8⁺T细胞（r=0.262，P < 0.01）、CD4⁺T细胞（r=0.12，P < 0.05）、巨噬细胞（r=0.252，P < 0.01）、中性粒细胞（r=0.242，P < 0.01）和树突状细胞（r=0.162，P < 0.01）的免疫浸润水平呈正相关。蛋白相互作用网络分析发现，AKT1、SIRT1、EP300、BCL2L11、SOD2、SGK1、AKT2、CREBBP、CDKN1B、SMAD4等蛋白与FOXO3具有明显相互作用。结论：基于肿瘤基因数据库分析发现，FOXO3基因在膀胱癌组织中低表达，其高表达与患者预后不良相关；FOXO3表达与膀胱癌免疫细胞的浸润水平有关。     

血清谷胱甘肽过氧化物酶与晚期非小细胞肺癌患者临床病理指标及预后的相关性

目的：探讨血清谷胱甘肽过氧化物酶（GSH-PX）活性与晚期非小细胞肺癌（NSCLC）患者临床病理指标、放化疗联合治疗的疗效及预后的相关性。方法：收集174例2018年1—12月于河北工程大学附属医院肿瘤科初次确诊、未经抗癌治疗的晚期NSCLC患者作为研究对象,以80例健康体检者为对照,患者于放化疗联合治疗前采集外周血,分离血清。采用基因表达谱数据动态分析（GEPIA）数据库分析GSH-PX的编码基因GPX3在不同肿瘤组织中的表达情况;5',5-二硫硝基苯甲酸（DTNB）法检测GSH-PX活性,比较放化疗联合治疗前晚期NSCLC患者和对照组血清GSH-PX活性,并分析血清GSH-PX活性与晚期NSCLC临床病理指标、放化疗联合治疗疗效及预后的相关性。结果：GEPIA数据库分析显示33类肿瘤组织中,21类肿瘤组织的GPX3 mRNA表达低于正常组织,其中包括肺鳞癌和肺腺癌（P<0.05）;DTNB法检测结果显示晚期NSCLC患者血清GSH-PX活性显著低于健康对照组（P<0.05）;与年龄、性别、病理类型无明显相关（均为P>0.05）,与吸烟史和肿瘤大小呈负相关（r=-0.165、-0.267,均为P<0.05）,与卡氏（KPS）评分和病理分级呈正相关（r=0.295、0.228,均为P<0.05）;治疗有效患者和无效患者血清GSH-PX活性的差异无统计学意义（P>0.05）,但GSH-PX高活性组患者总生存期显著高于低活性组（P<0.05）,且呈正相关（r=0.238,P=0.000）。结论：晚期NSCLC患者血清GSH-PX活性降低,与患者吸烟史、KPS评分、肿瘤大小、病理分级和总生存期显著相关。GSH-PX可能参与了NSCLC的发生发展,可作为评估预后的指标。     

晚期非小细胞肺癌病理特征及放化疗联合治疗疗效与患者血清超氧化物歧化酶的关系

目的：探讨晚期非小细胞肺癌（NSCLC）病理特征、放化疗联合治疗疗效、总生存期与患者血清超氧化物歧化酶（SOD）活力的关系。方法：采用基因表达谱数据动态分析数据库（GEPIA）分析不同肿瘤组织SOD的表达情况。收集2018年1—12月河北工程大学附属医院肿瘤科初次确诊的174例晚期NSCLC患者作为研究对象，80例健康者作为对照，放化疗联合治疗前采集患者外周血，分离血清，WST-1法检测血清中SOD活性，根据实体瘤疗效评价标准（RECIST 1.1）评估疗效，追踪随访，分析晚期NSCLC临床病理特征、放化疗联合治疗疗效、总生存期与SOD活力的关系。结果：GEPIA分析显示33类肿瘤中，19类肿瘤组织SOD表达异常，其中肺鳞状细胞癌和肺腺癌组织SOD表达较对照组显著降低（P < 0.05）；WST-1检测结果显示晚期NSCLC患者血清SOD活力显著低于对照组（P < 0.05），不同年龄、性别、病理类型患者血清SOD活力无显著性差异（P > 0.05）；不同吸烟史及病理分级患者血清SOD有显著性差异（P < 0.05），吸烟者血清SOD活力显著低于无吸烟者（P < 0.05），低分化患者血清SOD活力显著低于中、高分化者（P < 0.05）；患者血清SOD活力与放化疗联合治疗疗效呈正相关（r=0.554，P < 0.05），治疗有效患者血清SOD活力显著高于无效患者（P < 0.05），且SOD高活力组患者总生存期显著高于低活力组（P < 0.05）。结论：NSCLC患者血清SOD活性显著降低，且受患者吸烟史、病理分级的影响，可能与放化疗联合治疗疗效和总生存期有关。血清SOD或可作为评估NSCLC临床疗效和判断预后的潜在指标，辅助NSCLC放化疗的疗效判定及预后评估。     

晚期非小细胞肺癌病理特征及放化疗联合治疗疗效与患者血清超氧化物歧化酶的关系

目的：探讨晚期非小细胞肺癌（NSCLC）病理特征、放化疗联合治疗疗效、总生存期与患者血清超氧化物歧化酶（SOD）活力的关系。方法：采用基因表达谱数据动态分析数据库（GEPIA）分析不同肿瘤组织SOD的表达情况。收集2018年1—12月河北工程大学附属医院肿瘤科初次确诊的174例晚期NSCLC患者作为研究对象，80例健康者作为对照，放化疗联合治疗前采集患者外周血，分离血清，WST-1法检测血清中SOD活性，根据实体瘤疗效评价标准（RECIST 1.1）评估疗效，追踪随访，分析晚期NSCLC临床病理特征、放化疗联合治疗疗效、总生存期与SOD活力的关系。结果：GEPIA分析显示33类肿瘤中，19类肿瘤组织SOD表达异常，其中肺鳞状细胞癌和肺腺癌组织SOD表达较对照组显著降低（P < 0.05）；WST-1检测结果显示晚期NSCLC患者血清SOD活力显著低于对照组（P < 0.05），不同年龄、性别、病理类型患者血清SOD活力无显著性差异（P > 0.05）；不同吸烟史及病理分级患者血清SOD有显著性差异（P < 0.05），吸烟者血清SOD活力显著低于无吸烟者（P < 0.05），低分化患者血清SOD活力显著低于中、高分化者（P < 0.05）；患者血清SOD活力与放化疗联合治疗疗效呈正相关（r=0.554，P < 0.05），治疗有效患者血清SOD活力显著高于无效患者（P < 0.05），且SOD高活力组患者总生存期显著高于低活力组（P < 0.05）。结论：NSCLC患者血清SOD活性显著降低，且受患者吸烟史、病理分级的影响，可能与放化疗联合治疗疗效和总生存期有关。血清SOD或可作为评估NSCLC临床疗效和判断预后的潜在指标，辅助NSCLC放化疗的疗效判定及预后评估。     

舌鳞状细胞癌组织中硫氧还蛋白和硫氧还蛋白还原酶-1的表达及其意义

目的：研究硫氧还蛋白（Trx）和硫氧还蛋白还原酶-1（TrxR1）在舌鳞癌组织中的表达及其临床意义。方法：收集28例舌鳞癌组织和10例癌旁上皮组织,采用免疫组织化学SP三步法检测Trx和TrxR1蛋白在两种组织中的表达水平,并分析其表达与临床病理特征的关系。结果：舌鳞状细胞癌组织中的Trx和TrxR1蛋白表达水平明显高于癌旁正常上皮组织,癌组织和癌旁正常组织的Trx IOD值分别为（2.84±0.34）×10⁸和（3.91±3.00）×10⁷（P < 0.01）,TrxR1 IOD分别为（1.88±0.29）×10⁸和（0.69±0.32）×10⁷（P < 0.05）;Trx和TrxR1蛋白表达水平在不同肿瘤大小、分化程度组间表达的差异均具有统计学意义（P均 < 0.05）;Trx和TrxR1表达水平之间呈正相关关系（r=0.504,P < 0.05）,且Trx和TrxR1高表达组与低表达组的生存率差异具有统计学意义（P < 0.05）。结论：硫氧还蛋白及其还原酶-1可能为舌鳞状细胞癌的药物治疗靶点和早期诊断、肿瘤筛查的重要临床指标。     

伴病理性乳头溢液的乳腺癌患者癌组织整合素β1的表达及意义

目的：探讨整合素β1在伴病理性乳头溢液乳腺癌患者癌组织中的表达特征,及其对患者生存和预后的影响。方法：选取2006年1月1日-2013年12月31日期间,甘肃省人民医院收治的112例以乳头溢液就诊的乳腺癌患者作为研究对象,选取同期以乳房肿块或体检发现异常而就诊的112例乳腺癌患者作为对照,采用免疫组织化学方法检测整合素β1与Her-2在癌组织中的表达的差异,并分析整合素β1对乳腺癌预后的影响。结果：伴病理性乳头溢液组整合素β1阳性表达率为14.29%（16/112）,低于对照组的25%（28/112）,差异有统计学意义（χ²=4.073,P=0.044）。生存分析发现,对照组总生存期明显低于伴病理性乳头溢液组（HR=1.773,P=0.012）;病理性乳头溢液组中整合素β1高表达患者的总生存期明显低于低表达者（HR=2.573,P=0.023）;伴病理性乳头溢液组整合素β1的表达与Her-2水平呈正相关关系（r=0.281,P=0.001）。Cox风险比例模型检验发现,淋巴结转移、组织学分级与整合素β1高表达是影响伴病理性乳头溢液组患者总生存期的3个独立危险因素（P < 0.01）。结论：整合素β1在乳腺癌组织中高表达可显著降低乳腺癌患者总生存期,且与乳腺癌的不良病理特点呈正相关。     

镉暴露致大鼠肾损伤的量效关系及其机制

目的：探讨不同剂量氯化镉（CdCl₂）染毒对大鼠肾损伤的量效关系及其机制。方法：雄性SD大鼠40只,随机分为0、1、3和5mg/kg CdCl₂染毒组,每天灌胃1次。连续染毒4周后检测血清尿素氮（BUN）和肌酐（SCr）水平评价肾功能;HE染色观察肾脏病理损伤;透射电镜观察肾组织超微结构变化;DHE及MitoSOX荧光探针检测肾组织中活性氧（ROS）含量变化;免疫印迹检测SOD1、SOD2、GPx-1、CAT、Bax、Bcl-2以及GRP78蛋白的表达。结果：与对照组比较,镉暴露后大鼠体质量及肾体比差异无统计学意义（P>0.05）;随CdCl₂剂量增加,血清BUN含量逐渐增加（r=0.463,P<0.05）;肾脏病理损伤进行性加重,出现肾小球肾小管肿胀,肾小管管腔狭窄、上皮细胞坏死脱落堆积于管腔、管腔内可见管型,肾间质充血,炎细胞浸润;超微结构观察显示,肾小管上皮细胞线粒体损伤,肿胀、变形、空泡样变程度逐渐加重;肾组织中ROS含量逐渐增加;抗氧化酶SOD2、GPx-1和CAT表达逐渐增加（r依次为0.854、0.975、0.918,P均<0.05）,SOD1表达逐渐减少（r=-0.987,P<0.05）;肾脏组织促凋亡蛋白Bax表达在0~3 mg/kg CdCl₂处理组逐渐增加（r=0.226,P<0.05）,抑凋亡蛋白Bcl-2表达逐渐减少（r=-0.85,P<0.05）,Bax/Bcl-2呈剂量依赖性升高（r=0.83,P<0.05）;同时,内质网应激标志蛋白GRP78的表达也随CdCl₂剂量增加而升高（r=0.913,P<0.05）。结论：不同剂量镉暴露致雄性SD大鼠肾脏损伤存在剂量效应关系,其作用机制可能是镉暴露导致氧化还原稳态失衡,从而引起氧化应激,进一步引起凋亡和组织损伤。     

Cholinesterase activity of human lung tumours varies according to their histological classification

The probable involvement of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in cancer and the relevance of cholinergic responses for lung cancer growth prompted us to study whether cholinesterase activity of human lung is altered by malignancy. Surgical pieces of non-small lung carcinomas (NSLC) and their adjacent non-cancerous tissues (ANCT) were analysed for AChE and BChE activities. AChE activity in adenocarcinoma (AC) was 7.80 +/- 5.59 nmol of substrate hydrolysed per min and per mg of protein (mU/mg), the same as in their ANCT (8.83 +/- 4.72 mU/mg; P = 0.823); in large cell carcinoma (LCC), 7.52 +/- 3.32 mU/mg, approximately 50% less than in their ANCT (15.39 +/- 5.66 mU/mg; P = 0.043); and in squamous cell carcinoma (SCC), 1.39 +/- 0.58 mU/mg, 80% less than in ANCT (6.08 +/- 2.88 mU/mg; P = 0.003). BChE activity was 5.85 +/- 3.20 mU/mg in AC and 9.56 +/- 3.38 mU/mg in ANCT (P = 0.022); 2.94 +/- 2.01 mU/mg in LCC and 6.50 +/- 6.63 mU/mg in ANCT (P = 0.068); and 4.49 +/- 2.30 mU/mg in SCC and ANCT 6.56 +/- 4.09 mU/mg (P = 0.026). Abundant AChE dimers and fewer monomers were identified in lung and, although their distribution was unaffected by cancer, the binding with concanavalin A revealed changes in AChE glycosylation between SCC and their ANCT. The fall in BChE activity affected all molecules, with a strong decrease of the amphiphilic tetramers. Western blotting revealed protein bands with the expected mass of the principal AChE subunits, and the deeper intensity of the protein signal in SCC than in healthy lung, in lanes loaded with the same units of AChE activity, supported an augment in the amount of AChE protein/unit of AChE activity in SCC. The increased availability of acetylcholine in neoplastic lung, resulting from the fall of cholinesterase activity, may enhance cholinergic signalling and contribute to tumour progression.     

应用PBTK/TD模型研究毒死蜱经口重复染毒幼年大鼠的毒代动力学及毒效应学

目的:应用PBTK/TD模型研究幼年大鼠经口重复暴露毒死蜱(CPF)的毒代动力学及毒效应学。方法:21日龄雌性SD大鼠按体质量随机分成对照组、1.0、2.5、5.0、10.0和15.0 mg/kg毒死蜱剂量组,每天1次,连续灌胃10 d,在第3、6、10和11天收集血清和大脑皮质,测定血清CPF和3,5,6-三氯-2-吡啶(TCP)浓度、血清和皮质的乙酰胆碱酯酶活力(AChE)活力;收集第3、6和11天的尿液,测定尿液TCP累积量。在幼年大鼠毒死蜱经口暴露PBTK/TD模型中输入暴露条件,模拟各个指标的时量变化曲线。结果:以24 h为1个周期,每次染毒后血清CPF和TCP浓度随时间的增加先上升后下降,与对照组间的差异具有统计学意义(P<0.05)。血清和皮质AChE活力随时间亦呈周期性变化,均随着染毒剂量的增加而减小,具有统计学意义(P<0.05);而在停止染毒后第1天,血清AChE活力显著恢复,具有统计学意义(P<0.05)。结论:毒死蜱在重复染毒时,毒代谢动力学与毒效应学指标均呈周期性变化,其持续暴露会使血清和大脑皮质AChE维持在一定的抑制水平,停止暴露后可显著恢复。     

Biochemical markers for Fischer rat leukemia in a cell transplant model

Cellular glucose-metabolizing enzymes and acetylcholinesterase (AChE) have been utilized as biochemical markers of mononuclear cell (MNC) leukemia maintained by serial cell transplantation in F344 rats. We have evaluated the sensitivity and reproducibility of these tumor markers in comparison to other diagnostic criteria of leukemia. Weanling rats were injected with 2 X 10(7) leukemic spleen MNC and sampled at 6, 35, 63, and 83 days. At 6 days, the glycolytic enzyme activities from spleen that decreased were believed to be residual activity from injected leukemic MNC. Glycolytic enzyme activities in spleen MNC were normal at 35 days and no changes in blood MNC enzyme activity occurred at 6 days or 35 days. At 63 days, prior to clinical evidence of leukemia, glucose-metabolizing enzymes from spleen MNC changed, and there were decreases in AChE from both blood and spleen MNC that progressively decreased at 83 days, when there was depressed body weight, splenomegaly, elevated WBC, depressed RBC, hypoglycemia, hyperbilirubinemia, and elevated serum enzyme levels. Separation of leukemic MNC from blood and spleen enhances sensitivity of cellular enzyme responses and provides a reproducible model to study biochemical markers correlated with severity of leukemia.     

Unusually high expression of N-acetylglucosaminyltransferase-IVa in human choriocarcinoma cell lines: a possible enzymatic basis of the formation of abnormal biantennary sugar chain.

Structural analysis of the sugar chains of human chorionic gonadotropin (hCG) has revealed that abnormal biantennary structures appear specifically on hCG in the urine of choriocarcinoma patients. However, the enzymatic and molecular mechanisms of the biosynthesis of abnormal biantennary sugar chains have not yet been elucidated. In this report, the enzyme activities and the expression levels of mRNAs of N-acetylglucosaminyltransferases (GnT)-I to -V, beta-1,4-galactosyltransferase, and alpha-mannosidase II in normal human placentae and three human choriocarcinoma cell lines were investigated. GnT-IV activities in choriocarcinoma cell lines were increased from 16- to 66-fold and GnT-III activity was increased from 15- to 25-fold as compared with those in human placentae, whereas other enzyme activities were not increased significantly. The mRNA expression levels generally correlated with their enzyme activities. Among the two GnT-IV genes found in human tissues only GnT-IVa gene was strongly expressed in the cancer cells: from three to seven times as much as in the normal tissue, whereas that of GnT-IVb remained constant. On the basis of these results, we proposed that ectopic expression of GnT-IVa gene should occur along with the malignancy of trophoblastic tissues, and that the increased GnT-IV activity should be the main cause of the formation of abnormal biantennary sugar chains in choriocarcinoma. A possible enzymatic basis of the biosynthesis of abnormal biantennary sugar chains is discussed.     

Acetylcholinesterase and butyrylcholinesterase activities in human thyroid cancer cells

The appearance and distribution of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) in 12 human thyroid cancers and three normal thyroids were examined by electron microscopic study with indirect thiocholine method. The demonstration of AChE and BuChE activities in only two of nine cases of follicular and papillary carcinoma examined and none of the three cases of medullary carcinoma shows that the cholinesterases are not specific enzymes for the thyroid tumors. In normal thyroid tissue samples examined, no activities of AChE and BuChE were detected. On ultrastructural level AChE reaction product was revealed in the perinuclear space, in the endoplasmic reticulum, and in the Golgi complex of some but not in all cells in less-differentiated regions of the tumors. In contrast to the distribution of AChE, no staining for BuChE was noted in the Golgi elements. Ultrastructural localization of AChE activity in the thyroid cancer cells corresponds exactly to the current understanding of glycoproteins synthesis and processing in normal cells. The authors postulate that the copy of AChE gene suppressed in normal thyroid epithelium cells may be expressed in some follicular thyroid carcinoma cells. Their hypothesis is logical on the basis of recent finding of a significant homology between AChE and thyroglobulin.     

Collagenolytic activity of cervix carcinoma]

We estimated collagenolytic activity of 10 invasive carcinomas of the cervix by means of a biological assay. Collagenolysis was found in 7 specimens. In order to detect the origin of the collagenolytic enzyme additional inhibition studies with ethylendiaminetetraacetate and normal human serum were performed. Normal human serum inhibited the enzymatic process by a mean percentage of 14.9%, EDTA up to 9.1%. D-penicillamine and 1.10-o-phenanthroline which inhibit collagenase specifically stopped the enzymatic activity of the tumor completely. According to our results it can be suggested that the collagenolytic activity of carcinoma of the cervix uteri is not derived from serum or from polymorphonuclear cells but is created by the tumor itself.     

大鼠一次性经皮染毒毒死蜱的毒代动力学和毒效应学研究

目的:研究大鼠一次性经皮染毒毒死蜱(CPF)的毒代动力学和毒效应学特征。方法:成年雌性SD大鼠按体质量随机分成3个剂量组(69.75、139.5、279 mg/kg)和1个对照组(0 mg/kg),皮下注射染毒,各组在给药后3、6、12、24、48、72 h共6个时间点收集大鼠血液和大脑皮质样本,在0～24、0～48和0～72 h连续收集尿液样本。测定指标包括血清CPF浓度,血清和尿液3,5,6-三氯-2-吡啶(TCP)浓度以及血清和大脑皮质乙酰胆碱酯酶(AChE)活性。结果:经皮染毒CPF剂量越大,大鼠血清CPF和TCP浓度越高,尿TCP排出量越大,血和皮质AChE活性抑制越大;血清CPF浓度在6 h达峰值,血清TCP浓度在12～24 h达峰值,血AChE活性在24 h最低,大脑皮质AChE活性在24～48 h最低。各剂量组血清CPF的半衰期为29～48 h,0～72 h曲线下面积(AUC)为31～110 h·μmol/L;血清TCP的半衰期为28～54 h,0～72 h AUC值为897～3 450 h·μmol/L。血AChE活性和大脑皮质AChE活性呈正相关(P<0.01),且血中的抑制率大于大脑皮质(P<0.01)。用DoseResp函数拟合血清CPF和AChE活性,呈现出剂量反应关系。结论:CPF外暴露剂量与大鼠体内CPF、TCP和AChE活性相关,内暴露水平和血AChE活性存在剂量效应关系;尿TCP排出量可作为机体暴露水平的生物标志;血AChE活性是大脑皮质AChE活性的有效生物标志。     

Proteinase activity in malignant human breast cancers and NMU mammary tumours of the rat

In subcellular fractions of human mammary tumours and NMU tumours of rats proteinase activity was studied by means of the synthetic substrates Bz-dl-arginin-4-nitroanilid (BAPNA) and Bz-dl-arginin-2-naphthyl-amid (BANA). Using the substrate BAPNA enzymatic activity was found to be highest in low speed particulate fractions, whereas in NMU tumours of rats the bulk of the activities could be observed in the high speed supernatant. The substrates BAPNA and BANA were cleavaged enzymatically in human mammary tumours at pH 7 and pH 6, respectively, while in rats the maximum turnover of both substrates changed at value of pH 6.5. Enzyme activity with BAPNA was proved to be resistant to alkaline preincubation in human breast cancer tissue only. On the other hand, the enzymatic cleavage of BANA was completely lost in human as well as in rat tumour specimens under these experimental conditions. It can be concluded from these results that both enzyme activities measured in human malignant mammary tumours, which are known for their invading activity, represent two different proteolytic enzymes with their maximum activity at neutral and acidic pH. Similar enzyme activities are quite different in NMU tumours, which are not invasive.     

Urinary 6 beta-hydroxycortisol/cortisol ratios measured by high-performance liquid chromatography for use as a biomarker for the human cytochrome P-450 3A4.

Cortisol is metabolized to 6 beta-hydroxycortisol by the human cytochrome P-450 3A4, an enzyme implicated in the critical epoxidation reactions of aflatoxin B1 and certain polycyclic aromatic hydrocarbons. A method has been developed to measure the 6 beta-hydroxycortisol:cortisol ratio in human urine by reverse-phase high-performance liquid chromatography using diode-array detection. This method permits measurement of both cortisols in a single chromatographic run and has the potential to measure other metabolites useful for the determination of other human cytochrome P-450 activities. Using the 6 beta-hydroxycortisol:cortisol ratio as a marker of cytochrome P-450 3A4 activity adjusts for the circadian fluctuation in cortisol levels, providing a determination that is consistent over a 24-h period, obviating the need for 24-h urine collections. This high-performance liquid chromatography technique was used for the analysis of 14 different human urines and a range of ratios of 1.6 to 21.7 (mean +/- SE, 6.2 +/- 1.6) was found. These values are consistent with those reported in the literature. This method should be useful in molecular epidemiological studies investigating relationships between the susceptibility from environmental carcinogen exposure and cancer risk.