外周血淋巴细胞替代肺癌细胞体外药敏实验的相关性研究

目的本实验通过肺癌患者的外周血淋巴细胞和肺癌细胞的体外培养,对临床上常用的四种化疗药物卡铂、长春新碱、五氟尿嘧啶、环磷酰胺进行药物敏感性检测,并检验两者的相关性。探讨用患者自身外周血淋巴细胞的药敏实验结果替代肺癌细胞药敏实验结果的可行性。方法对60例术前未经任何治疗的非小细胞肺癌患者的肺癌细胞和自身外周血淋巴细胞进行体外原代培养。培养48h后行四氮唑盐（MTT）比色法检测这些细胞对卡铂、5-氟脲嘧啶、环磷酰胺、长春新碱等临床常用化疗药物的敏感性。结果同一种化疗药物卡铂、长春新碱、五氟尿嘧啶、环磷酰胺对患者肺癌细胞和自身外周血淋巴细胞的抑制率之间具有良好的,立相关性。相关系数分别为：0．701、0．668、0．543、0．696,且有统计学意义（P〈0．05）。根据60例肺癌细胞和自身外周血淋巴细胞对化疗药物的敏感率（敏感率=敏感病例数／病历总数×100％）来分析：患者肺癌细胞和自身外周血淋巴细胞对相同化疗药物卡铂、长春新碱、五氟尿嘧啶、环磷酰胺的敏感率无显著性差异,x2值分别为0．786、1．645、0．263、0．261,且均无统计学意义（P〉0．05）。结论肺癌患者的外周血淋巴细胞和肺癌细胞对相同的化疗药物敏感性无显著性差异,两者之间具有良好的正相关性。当肺癌患者已失去手术机会、或由于各种原因实体瘤标本无法取得时可以用外周血淋巴细胞对化疗药物的敏感性替代肿瘤细胞进行药物筛选。     

肝癌外周血淋巴细胞和肿瘤细胞多药耐药基因表达的研究

目的探讨肝癌患者外周血淋巴细胞和肿瘤细胞多药耐药糖蛋白(P-gp)表达的相关性。方法选取肝癌患者38例,应用流式细胞免疫荧光技术检测肿瘤细胞和外周血淋巴细胞多药耐药糖蛋白的表达。结果38例肝癌肿瘤细胞P-gp阳性细胞率为(2.9%±0.9)%,外周血淋巴细胞P-gp阳性细胞率为(2.4%±0.8)%,肝癌肿瘤细胞与外周血淋巴细胞P-gp表达阳性率差异无统计学意义(P>0.05)。结论肝癌患者可以应用外周血淋巴细胞代替肿瘤细胞进行多药耐药基因表达的检测,以提高药敏预测的准确性。     

板蓝根乙酸乙酯部位体外抗病毒活性研究

目的对板蓝根的乙酸乙酯部位各化学组分进行体外抗单纯疱疹病毒Ⅰ型(HSV1)活性研究。方法利用溶剂法和色谱法从板蓝根中提取分离出20个乙酸乙酯组分,利用四甲基偶氮唑盐比色(MTT)法检测这20个组分体外对HSV1病毒的抑制作用,以细胞存活率、治疗指数(TI)和细胞保护率为评价指标,比较各化学组分的体外抗病毒的效果。结果板蓝根乙酸乙酯8个组分均有明显的抗HSV1病毒作用,对病毒的抑制率均远大于50%。结论板蓝根乙酸乙酯部位中存在直接抗病毒较强的活性成分,对其结构进行深入研究,将有助于阐明板蓝根抗病毒药效物质基础。     

应用MTT法对恶性肿瘤原代细胞培养进行规范化体外药敏试验的研究

目的探究应用四甲基偶氮唑盐(MTT)比色法对恶性肿瘤原代细胞培养进行规范化体外药敏试验的可行性和实用性。方法30例恶性肿瘤患者的新鲜肿瘤组织,进行原代细胞培养纯化。采用MTT比色法规范化检测12种肿瘤化疗药物对人恶性肿瘤原代细胞培养进行体外药敏试验。结果紫杉醇、多西他赛对胃癌和结直肠癌原代培养细胞的药物敏感性均为77.78%,而对乳腺癌原代培养细胞的药物敏感性为88.89%;阿霉素对结直肠癌、胃癌以及乳腺癌原代培养细胞的药物敏感性依次为66.67%、55.56%、55.56%;表阿霉素依次为77.78%、77.78%、66.67%;环磷酰胺和顺铂依次为66.67%、88.89%、77.78%;卡铂依次为55.56%、77.78%、77.78%;奥沙利铂依次为77.78%、88.89%、88.89%;氟尿嘧啶依次为77.78%、66.67%、55.56%;吉西他滨依次为66.67%、77.78%、66.67%;长春瑞滨依次为77.78%、66.67%、66.67%;依托泊苷依次为66.67%、77.78%、77.78%。结论MTT法对于临床合理用药,实现肿瘤患者的个体化化疗具有重要的价值。     

老年肺癌患者肿瘤细胞和外周血淋巴细胞体外化疗药敏试验的相关性

目的 探讨老年非小细胞肺癌患者体外肿瘤细胞和外周血淋巴细胞对化疗药物敏感性的关系.方法 选取52例老年非小细胞肺癌患者,采用MTT法体外药敏试验,检测外周血淋巴细胞和肿瘤细胞对13种化疗药物的敏感性.结果 老年非小细胞肺癌患者外周血淋巴细胞对顺铂(CDDP)、卡铂(CBP)、奥沙利铂(LO HP)、紫杉醇(PTX)、环磷酰胺(CTX)、异环磷酰胺(ICTX)、吡喃阿霉素(THP)、依托泊苷(VP-16)、吉西他滨(GEM)、长春新碱(VCR)、长春瑞滨(NVB)等11种化疗药物的敏感性与非小细胞肺癌肿瘤细胞的药敏结果有相关性.而对阿霉素、羟基喜树碱这2种化疗药物的敏感结果无相关性.结论 老年非小细胞肺癌患者可以应用外周血淋巴细胞代替肿瘤细胞进行体外化疗药物敏感试验.     

盐酸多巴酚丁胺注射液细胞毒性研究

目的：比较不同来源盐酸多巴酚丁胺注射液体外细胞毒性的差别,为遴选优质药品、保障用药安全提供科学依据.方法：将不同来源国产盐酸多巴酚丁胺注射液原液以及稀释液,与L929细胞接触培养,通过倒置相差显微镜观察其形态,采用MTT法量化细胞毒性,计算相对增值率和IC50值,进行细胞毒性评价.并将国产盐酸多巴酚丁胺注射液与进口制剂进行比较.结果：8个厂家生产的盐酸多巴酚丁胺注射液原液及进口产品的细胞毒性分级均为4级.不同企业生产的盐酸多巴酚丁胺注射液的IC50值差异较大.盐酸多巴酚丁胺注射液C和F的IC50值较小,说明这两个企业产品的细胞毒性较其他样品大.进口盐酸多巴酚丁胺注射液的IC50值为99.9 mg·L-1,较其他样品高,说明其细胞毒性较其他样品低.进口盐酸多巴酚丁胺注射液中仅添加亚硫酸氢钠一种辅料,国产盐酸多巴酚丁胺注射液中添加的辅料主要为依地酸二钠、亚硫酸氢钠、盐酸半胱氨酸等.按各辅料在国产盐酸多巴酚丁酸注射液中的浓度测其细胞毒性,依地酸二钠、盐酸半胱氨酸和亚硫酸氢钠的细胞毒性分级均为3级.将依地酸二钠的浓度稀释为0.03 mg·ml-时,其细胞毒性分级为1级,无细胞毒性.将盐酸半胱氨酸稀释到0.11 mg·ml-1时,其细胞毒性分级为1级,基本无细胞毒性.结论：不同厂家生产的盐酸多巴酚丁胺注射液由于生产条件不同、处方不同等原因,这些产品的细胞毒性存在较大的差别.与进口制剂比较,国产制剂中添加的辅料浓度较大,可适当降低其浓度,以减少药物制剂的细胞毒性,从而提高药品的安全性.     

美洲大蠊提取物对3株消化系统肿瘤细胞的细胞毒性研究

目的探讨美洲大蠊提取物的抗肿瘤作用。方法通过聚酰胺柱层析分离划段得到美洲大蠊醇提物的不同部位,采用四甲基偶氮唑盐比色法（MTT法）对所得部位进行体外肿瘤细胞毒性测试。结果多个部位样品的半数抑制浓度（IC50）值小于10μg／mL。结论美洲大蠊提取物中确实存在具有肿瘤细胞生长抑制作用的物质,值得进一步研究。     

胃腺癌细胞对5-氟尿嘧啶与奥沙利铂的敏感性研究

目的探讨不同分化程度的胃腺癌细胞对5-氟尿嘧啶和奥沙利铂的敏感性。方法以不同浓度的5-氟尿嘧啶和奥沙利铂分别处理低分化和中分化胃腺癌细胞,采用四甲基偶氮唑盐比色法（MTT法）检测药物对胃腺癌细胞增殖的影响。结果5-氟尿嘧啶和奥沙利铂分肿对2种胃腺癌细胞的增殖均具有显著的抑制作用,且随药物浓度增加而增强,中分化的胃腺癌细胞对2种化疗药物的敏感性均明显高于低分化的胃腺癌细胞。结论不同浓度的5-氟尿嘧啶和奥沙利铂对胃腺癌细胞的抑制作用不同,不同分化程度的胃腺癌细胞对5-氟尿嘧啶和奥沙利铂的敏感性也有不同。     

牛和羊胎盘肽的制备及其生物活性的研究

目的探索一种新型的免疫调节剂,充分利用动物胎盘。方法利用超滤法从牛和羊胎盘中提取胎盘肽,采用改良的加脲Tricine-SDS-PAGE法测定其相对分子质量;利用紫外光扫描其最大吸收峰;并通过建立体外抑制兔淋巴细胞模型,利用四甲基偶氮唑盐(MTT)比色法测定所提取胎盘肽的免疫调节作用。结果从牛和羊胎盘中成功提取出了胎盘肽,其相对分子质量分别为4 785和4 386;其最大吸收峰分别为210.5和225.4 nm;牛和羊胎盘肽均可使顺铂抑制的兔外周血T淋巴细胞的转化率显著提高,其中牛胎盘肽以1∶1×103、1∶1×1042组吸收度明显高于抑制组(P<0.05);羊胎盘肽以1∶1×102、1∶1×103和1∶1×1043组效果最为明显(P<0.01)。结论牛和羊胎盘中含有的胎盘肽可成功提取出来,其可明显提高淋巴细胞的转化率,是一种较为理想的免疫调节剂。     

替莫唑胺联合分子靶向药物抗人脑胶质瘤细胞的体外试验研究

<正>现今化疗药物种类繁多,但有效率均不理想,本实验应用四甲基偶氮唑盐微量酶反应比色法(MTT)法比较替莫唑胺(TMZ)联合分子靶向药物西妥昔单抗(C225)对人脑胶质瘤细胞的抑制作用,旨在探索一种有效、新型的化疗方案。     

P-糖蛋白表达在食管贲门癌中的临床意义

目的 探讨P-糖蛋白(P-gp)在食管贲门癌中的表达及临床意义.方法 采用流式细胞免疫荧光法检测P-gp 在32例食管鳞癌和32例贲门腺癌及相应癌旁组织中的表达.分析P-gp的表达与肿瘤浸润深度、淋巴结转移之间的关系.结果 P-gp在食管癌中的阳性表达率为85%,贲门癌中为65%,并且P-gp在食管贲门癌组织的表达均高于相应的癌旁组织(P＜0.01).P-gp的表达与淋巴结转移有关(P＜0.01),而与肿瘤组织浸润深度无关(P＞0.05).结论 食管贲门癌组织中P-gp有先天性高表达,这是造成食管癌化疗效果差的一个重要的原因.P-gp的表达可在一定程度上反映肿瘤淋巴结转移的生物学特征.     

盐酸埃他卡林对大鼠血压及心脏功能的影响

盐酸埃他卡林是军事医学科学院毒物药物研究所自行设计合成的脂肪仲胺类化合物 ,动物实验表明它具有选择性抗高血压作用 ,即对清醒正常血压大鼠的血压影响较轻 ,对清醒高血压大鼠的血压影响明显 ,降压强度与动物血压高低相关。传统的降压作用机制不能解释其抗高血压作用 ,进一步的研究证明盐酸埃他卡林抗高血压作用的机制是作用于ＡＴＰ敏感性Ｋ+通道 ,使细胞膜发生超极化 ,降低细胞内的钙离子浓度 ,而导致血管扩张 ,血压下降 ,属ＡＴＰ敏感性Ｋ+通道开放剂。为进一步了解盐酸埃他卡林心血管作用的特点 ,本文在自发性高血压大鼠及麻醉的Ｗｉ…     

In vitro investigation of the genotoxic and cytotoxic effects of thiacloprid in cultured human peripheral blood lymphocytes

Thiacloprid, a neonicotinoid insecticide, is widely used for controlling various species of pests on many crops. The potential genotoxic effects of thiacloprid on human peripheral blood lymphocytes (PBLs) were investigated in vitro by the chromosome aberrations (CAs), sister chromatid exchanges (SCEs), and cytokinesis‐block micronucleus (MN) assays. The human PBLs were treated with 75, 150, and 300 μg/mL thiacloprid in the absence and presence of an exogenous metabolic activator (S9 mix). Thiacloprid increased the CAs and SCEs significantly at all concentrations (75, 150, and 300 μg/mL) both in the absence and presence of the S9 mix and induced a significant increase in MN and nucleoplasmic bridge formations at all concentrations for 24 h and at 75 and 150 μg/mL for 48‐h treatment periods in the absence of the S9 mix; and at all concentrations in the presence of the S9 mix when compared with the control and solvent control. Thiacloprid was also found to significantly induce nuclear bud (NBUD) formation at 300 μg/mL for 24 h and at 150 μg/mL for 48‐h treatment times in the absence of the S9 mix and at the two highest concentrations (150 and 300 μg/mL) in the presence of the S9 mix. Thiacloprid significantly decreased the mitotic index, proliferation index, and nuclear division index for all concentrations both in the absence and presence of the S9 mix. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 631–641, 2014.     

镍,铅,汞化合物体外诱发人淋巴细胞（UDS）的检测

用人外周血淋巴细胞的非程序ＤＮＡ合成对镍、铅、汞化合物进行了测试，结果表明，这三种重金属都具有诱发ＵＤＳ的活性，并提示，受试重金属化合物达到一定浓度后，ＵＤＳ的ｃｐｍ值不再随剂量增高而增高，似乎在一定实验条件下ＵＤＳ的修复合成能力有一定限度。     

The phytoestrogens coumoestrol and genistein induce structural chromosomal aberrations in cultured human peripheral blood lymphocytes

The clastogenic potential of the phytoestrogens coumoestrol (COUM), genistein (GEN) and daidzein (DAI) has been studied in human peripheral blood lymphocytes in vitro. After exposure of the cultured lymphocytes to 50 or 75 μM COUM or 25 μM GEN for 6 h, a clear induction of structural chromosomal aberrations was observed by cytogenetic analysis. The major alterations were chromatid breaks, gaps and interchanges. In contrast, DAI did not induce chromosome aberrations even at 100 μM. These results, together with previously published reports on the induction of micronuclei and DNA strand breaks in cultured Chinese hamster V79 cells by COUM and GEN, but not DAI, suggest that some but not all phytoestrogens have the potential for genetic toxicity. Received: 13 August 1998 / Accepted: 26 October 1998     

生长抑素增强胆囊癌细胞化疗敏感性的研究

目的 :探讨在生长抑素预先作用下 ,胆囊癌细胞化疗敏感性的变化。方法 :运用生长抑素预先作用于体外培养的人胆囊癌细胞株 ,2 4h后加入梯度浓度的化疗药物阿霉素 ,观察癌细胞的生长曲线 ,并与单独使用化疗药物的癌细胞生长曲线相比较。结果 :阿霉素能够明显抑制胆囊癌细胞的生长 ,其作用呈现效应 -浓度依赖性 (P <0 .0 5)。生长抑素能够诱导离体胆囊癌细胞生长停滞于DNA合成期 (S期)。经生长抑素预处理后 ,阿霉素对癌细胞生长的抑制效果较单独用药为佳 (P <0 .0 1 )。结论 :生长抑素可增强胆囊癌细胞对化疗药物阿霉素的敏感性。     

地塞米松对溃疡性结肠炎鼠外周血及肠黏膜固有层淋巴细胞凋亡的影响

目的观察溃疡性结肠炎(UC)大鼠外周血淋巴细胞(PBL)及肠黏膜固有层淋巴细胞(LPL)凋亡状态,并探讨地塞米松(Dex)对其的影响。方法将45只大鼠随机分为正常组、模型组、实验组3组,各15只,模型组及实验组用三硝基苯磺酸(TNBS)/乙醇灌肠制作实验性溃疡性结肠炎的动物模型,正常组以生理盐水灌肠作为对照,分离各组大鼠PBL及LPL,植物血凝素(PHA)刺激培养72h,流式细胞仪检测其凋亡率,实验组的PBL及LPL再经Dex作用后检测上述指标。结果模型组大鼠LPL在PHA刺激培养72h后凋亡率(5.89±0.65)%较正常组(17.14±1.95)%明显降低,PBL凋亡率(5.28±0.43)%较正常组(6.34±0.71)%两组间无明显差异,实验组PBL(10.85±1.04)%及LPL(15.89±0.65)%凋亡率较模型组增高。结论 UC鼠LPL凋亡减少,而PBL无明显凋亡异常,LPL凋亡减少可能同溃疡性结肠炎发病机制有关,地塞米松通过促进LPL凋亡从而达到治疗作用。     

The genotoxic and antigenotoxic effects of Salvia fruticosa leaf extract in human blood lymphocytes

Abstract

The aim of this study was to investigate the genotoxic and antigenotoxic effects of Salvia fruticosa (Sf) leaf extract with the absence and presence of S9 mix using sister chromatid exchange (SCE), chromosome aberration (CA) and micronucleus (MN) formation test systems in human peripheral blood lymphocytes (HPBLs) that were treated with 1.5-, 3.0- and 6.0-µL/mL concentrations for 24- and 48-hour treatment periods. The cytotoxicity of Sf leaf extract was also investigated by calculating the mitotic index (MI), proliferation index (PI) and nuclear division index (NDI). In the absence of S9 mix, Sf leaf extract alone increased SCE frequency at the 48-hour treatment period; however, it induced the CA and MN at all concentrations and at all treatment periods. Sf plus MMC (mitomycin C) synergically induced SCE and CA, except the highest concentration of Sf leaf extract and MMC on induction of SCE. In addition, Sf leaf extract induced the effect of MMC on MN frequency for 24 hours, but it significantly decreased the effect of MMC on MN frequency for the 48-hour treatment period. Sf leaf extract showed a cytotoxic effect by decreasing the MI; however, it did not decrease the PI and NDI. In the presence of S9 mix, Sf leaf extract did not increase the SCE, when compared to solvent control, whereas it reduced the effect of cyclophosphamide (Cyp). Sf leaf extract induced the CA and MN, but could not increase the effect of Cyp on CA and MN formation. Sf leaf extract had no cytotoxic effect; however, it induced the cytotoxicity of Cyp.     

Jatropha curcas leaf and bark fractions protect against ultraviolet radiation-B induced DNA damage in human peripheral blood lymphocytes

The present study is conducted to investigate the antioxidant potential of Jatropha curcas root bark extract (RB4 fraction) and leaf extract (L1 fraction), and to study their effects on UVB-radiation-induced DNA damage in cultured human blood lymphocytes. In this study, J. curcas showed strong antioxidant property in different free radical scavenging systems. Both the fractions effectively scavenged hydroxyl (OH), superoxide anion (O₂^–), 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid radical cation (ABTS⁺) in a concentration-dependent manner. The IC₅₀ (Inhibitory Concentration 50) values of J. curcas fractions were compared to standard ascorbic acid used in this study. The antioxidant potential of a compound was directly proportional to the photoprotective effect. In this study, human peripheral blood lymphocytes (HPBL) were exposed to UVB-radiation and there was an increase in comet attributes (% tail DNA, tail length, tail movement and Olive tail moment). Jatropha curcas RB4 fraction and L1 fraction treatment before UVB-irradiation significantly decreased the % tail DNA, tail length, tail moment and Olive tail moment in irradiated HPBL. These results suggested that J. curcas exhibited strong antioxidant property and RB4 and L1 fractions protected UVB-radiation-induced DNA damage in HPBL.