mdr1启动子调控CD∷UPP基因对紫杉醇耐药卵巢癌细胞的杀伤作用

目的:探讨腺病毒介导的mdr1启动子调控胞嘧啶脱氨酶∷尿嘧啶磷酸核糖转移酶(CD∷UPP)融合基因联合5-氟胞嘧啶(5-FC)对紫杉醇耐药卵巢癌细胞的特异性杀伤作用。方法:扩增、纯化含有mdr1-CD∷UPP基因的重组腺病毒,转染人卵巢癌紫杉醇耐药细胞株A2780/Taxol和亲本细胞株A2780,RT-PCR检测mdr1和CD∷UPP基因的表达水平;之后加入5-FC,MTT法检测细胞抑制情况及旁观者效应,并观察腺病毒转染后裸鼠移植瘤的生长情况。结果:mdr1和CD∷UPP基因在A2780/Taxol细胞中可稳定表达,转染后A2780/Taxol组的细胞生长明显低于A2780组;转基因的A2780/Taxol细胞联合5-FC后可通过旁观者效应杀伤周围未转基因的耐药细胞;耐药组移植瘤生长明显受到抑制,肿瘤体积为(569.10±187.93)mm3,对照组肿瘤体积为(2111.98±230.82)mm3,差异有统计学意义(P<0.01)。结论:mdr1启动子可调控CD∷UPP基因特异性表达并特异性杀伤紫杉醇耐药卵巢癌细胞。     

TB-Ab、ADA、和CEA对良恶性胸水鉴别诊断价值

[目的]探讨胸水结核抗体(TB-Ab-IgG)、腺苷脱氨酶(ADA)、癌胚抗原(CEA)联合检测对良恶性胸腔积液的鉴别诊断价值。[方法]采用斑点金免疫渗滤试验(DIGFA)、酶连续监测法和酶联免疫(ELISA)双抗体夹心法对119例胸腔积液患者行胸水/血清TB-Ab-IgG、ADA和CEA检测分析。[结果]97例结核性胸膜炎患者胸水、血清中TB-Ab-IgG的阳性率分别为61.9%(60/97)和70.1%(68/97),特异性分别为90.1%(20/22)和86.4%(19/22)。ADA活性在结核性和癌性胸腔积液中分别为(59.58±29.85)U/L和(15.31±7.36)U/L(P﹤0.01)。以P-ADA﹥40 U/L做为诊断结核的临界值,其敏感性为79.3%,特异性为86.4%;以P-ADA/S-ADA﹥1为临界值,其敏感性为97.7%,特异性为95.5%。CEA在结核性和癌性胸腔积液中的阳性率分别为8.20%和63.6%,特异性91.8%(89/97)。[结论]胸水和血清TB-Ab-IgG、ADA、CEA联合检测对良恶性胸腔积液具有诊断与鉴别诊断价值。     

多种雷公藤单体抑制HL-60细胞腺苷脱氨酶活性

目的研究多种雷公藤单体对HL-60细胞腺苷脱氨酶（ADA）的影响和诱导细胞凋亡作用。方法分别用雷公藤春碱、雷公藤内酯甲、雷公藤乙素、雷公藤黄素、雷公藤呋喃南蛇碱、雷公藤定碱（10mg／L、1mg／L、0．1mg／L）与HL-60细胞共孵育24h后测定ADA活性，选出抑制作用最强的雷公藤内酯甲。将HL-60细胞与雷公藤内酯甲10mg／L、1mg／L、0．1mg／L以及0．1mg／L雷公藤内酯甲＋腺苷0．5mmol／L、腺苷0．5mmol／L共孵育24h后测定细胞凋亡率。结果①雷公藤内酯甲和雷公藤呋喃南蛇碱均能明显抑制HL-60细胞ADA活性（P〈0．01），抑制作用与浓度成正比；雷公藤春碱仅在10mg／L浓度时有明显的ADA抑制作用（P〈0．01）；雷公藤黄素、雷公藤定碱与雷公藤乙素无明显ADA抑制作用（P〉0．05）。②HL-60细胞与雷公藤内酯甲10mg／L、1mg／L作用均可见明显的凋亡，单加内酯甲0．1mg／L时或单加腺苷时与对照组比较凋亡率无显著差异，但同时再加入腺苷有很高的凋亡率，两者有协同作用。结论雷公藤内酯甲、雷公藤呋喃南蛇碱和雷公藤春碱等雷公藤单体在一定浓度下对HL-60细胞ADA有抑制作用，可能是ADA一种抑制剂。雷公藤内酯甲在抑制细胞ADA活性的同时也造成明显的细胞凋亡。     

腺苷脱氨酶在结核性胸膜炎中的临床意义

目的探讨腺苷脱氨酶（ADA）在结核性胸膜炎中的临床意义。方法测定32例结核性胸膜炎患者与28例恶性胸腔积液患者的胸水、血液、支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）的水平。结果结核性胸膜炎患者的胸水腺苷脱氨酶（ADA）平均值为（46．68&#177;1．64）u／L，恶性胸腔积液患者的胸水腺苷脱氨酶（ADA）平均值为（24．76&#177;2．34）u／L，两者相比差异有显著性P〈0．01，结核性胸膜炎患者的血液腺苷脱氨酶（A-DA）平均值为（23．46&#177;1．28）u／L，恶性胸腔积液患者的血液腺苷脱氨酶（ADA）平均值为（22．76&#177;1．34）u／L，两者相比差异无显著性（P〉0．05），结核性胸膜炎患者的支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）平均值为（22．56&#177;1．17）u／L。恶性胸腔积液患者的支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）平均值为（21．96&#177;1．85）u／L，两者相比差异无显著性（P〉0．05），结核性胸膜炎患者的胸水腺苷脱氨酶（ADA）／血液腺苷脱氨酶（ADA）平均值（A1）为（1．82&#177;0．26），结核性胸膜炎患者的胸水腺苷脱氨酶（ADA）／支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）平均值（A2）为（1．78&#177;0．29），两者相比差异无显著性（P〉0．05），恶性胸水患者的胸水腺苷脱氨酶（ADA）／血液腺苷脱氨酶（ADA）平均值（B1）为（1．02&#177;0.16），恶性胸膜炎患者的胸水腺苷脱氨酶（ADA）／支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）平均值（B2）为（0．98&#177;0．26），两者相比差异无显著性（P〉0．05），但A1与B1，A2与B2相比差异有显著性（P〈0．01）。结论腺苷脱氨酶对诊断结核性胸膜炎有特异性，胸水腺苷脱氨酶（ADA）／血液腺苷脱氨酶（ADA）平均值（A1），胸水腺苷脱氨酶（ADA）／支气管肺泡灌洗液（BALF）中腺苷脱氨酶（ADA）的平均值（A2）在1．8左右有一定的临床价值。     

hTERT启动子调控腺病毒介导的CD基因治疗卵巢癌的实验研究

目的:构建人端粒酶逆转录酶(hTERT)启动子驱动的含胞嘧啶脱氨酶(CD)基因的复制缺陷型腺病毒载体,使CD基因只能在端粒酶阳性的细胞表达.方法:采用细胞内同源重组法构建出hTERT启动子调控的携带CD基因的复制缺陷型腺病毒Ad-hTERTp-CD,经PCR鉴定正确后进行扩增、纯化和滴度测定.分别将腺病毒感染人卵巢癌细胞株SKOV3及人胚肺成纤维细胞,MTT法检测受染细胞的存活率.结果:构建的重组腺病毒中带有hTERT启动子及CD基因.用不同滴度的重组腺病毒以及不同浓度的5-FC作用于SKOV3细胞后,MTT法检测到细胞的存活率随着病毒滴度和5-FC浓度的增加而不断降低;而同样的处理对人胚肺成纤维细胞的也有部分杀伤作用,但远较SKOV3细胞弱,差异有统计学意义(P＜0.01).结论:本实验构建的由hTERT启动子调控的携带CD基因的复制缺陷型腺病毒Ad-hTERTp-CD对SKOV3细胞具有靶向杀伤的能力.     

Effects of garlic and black grape extracts on the activity of adenosine deaminase from cancerous and noncancerous human urinary bladder tissues

Aim Possible effects of garlic (Allium sativum) and black grape (Fructus vitis minuta) with known antioxidant potential on adenosine deaminase (ADA) activities were investigated in cancerous and noncancerous human bladder tissues. Methods The effects of garlic and black grape extracts on adenosine deaminase (ADA) activities were measured in 20 pairs of cancer and adjacent normal human bladder tissues with and without pre-incubation with garlic and black grape extracts at different concentrations. Results No significant difference was observed between ADA activities in cancerous and non cancerous bladder tissues without plant extract (5.85 ± 3.78 versus 7.63 ± 2.88, respectively). At the 1/3 and 1/1 plant extract/bladder tissue homogenate ratios, the garlic extract completely abolished the ADA activity in both cancerous and noncancerous tissues. At the 1/3 plant extract/bladder tissue homogenate ratio, the black grape extract decreased the activity significantly as compared to without extract (2.01 ± 1.30 versus 5.85 ± 3.78; p < 0.05 for cancerous tissue and 2.10 ± 1.66 versus 7.63 ± 2.88; p < 0.05 for noncancerous tissue) and, at the 1/1 plant extract/bladder tissue homogenate ratio, completely abolished the activity. Conclusion Our results show that garlic and black grape have potential to inhibit ADA activity in both cancerous and adjacent normal human bladder tissues.We suggest that this might be rational basis for the uses of garlic and black grape in the complementary therapy of urinary bladder cancer.     

联检TB-DNA、LAM-IgG和ADA诊断小儿结核性脑膜炎

目的评价联检脑脊液结核杆菌DNA(TB-DNA)、抗阿拉伯糖甘露糖脂IgG抗体(LAM-IgG)和腺苷脱氨酶(ADA)指标诊断小儿结核性脑膜炎(结脑)及评价病情的价值。方法结脑组67例、非结核组38例,脑脊液标本应用聚合酶链反应法检测AFB、斑点免疫金渗滤试验法检测LAM-IgG、酶动力学终点法检测ADA。结果TB-DNA LAM-IgG诊断结脑的特异性为100%、阳性率为86.1%。结脑病情进展,ADA呈升高趋势;病情好转,ADA呈下降趋势。结论在诊断结脑方面,TB-DNA LAM-IgG联合试验的应用价值明显高于经典方法;在动态了解病情方面,ADA有较高的应用价值。三者联合应用可提高对小儿结脑诊断和评价的价值。     

Modulation of the Effect of l-β-D-Arabinofuranosylcytosine by 6-Mercaptopurine in L1210 Cells

In L1210 cells incubated with l- β -D-arabinofuranosylcytosine (ara-C), 6-mercaptopurine (6-MP) significantly potentiated 1- β -D-arabinofuranosylcytosme 5'-triphosphate (ara-CTP) accumulation and ara-C incorporation into DNA (ara-C/DNA). The cytotoxicity of these two drugs was assessed to be at least additive by clonogenic assay. l- β -D-Arabinofuranosylnracil (ara-U) level in a cell suspension was suppressed by 6-MP in a concentration-dependent fashion, though intracellular cytidine deaminase (CDD) activity was not affected by 6-MP. In addition, extracted CDD activity was not directly inhibited by 6-MP or by its intracellular metabolites in vitro. After preincubation in the presence or absence of 6-MP, the cell suspension was fractionated to obtain the spent medium and cell pellet. Then, each fraction was incubated with ara-C. Ara-U formation in the spent medium was found to increase conspicuously in relation to the time of preincubation in the control and it was suppressed by 6-MP pretreatment. Ara-U formation in the cell compartment increased slightly in relation to the time of preincubation in the control and substantially no suppression of ara-U formation was observed in spite of 6-MP pretreatment. In conclusion, intracellularly synthesized CDD was thought to be rapidly shed into the medium and the released CDD could play an important role in ara-C inactivation. 6-MP interrupted some step between synthesis and shedding of CDD, resulting in a decrease of the ara-C deamination in the medium and enhancement of its antileukemic effect.     

ADA和CEA联合检测在鉴别良恶性胸水中的价值

本文对癌性和结核性胸膜炎各30例进行了ADA、CEA测定.肺癌组ADA值(13.3±4.0U/L)明显低于结核组(59.6±33.6U/L)(P<0.01).CEA测定的阳性率分别为100.0％和3.3％,二组差异有显著性(P<0.01).     

Insulin-dependent diabetes mellitus and severe atopic dermatitis in a child with adenosine deaminase deficiency

We report a 2.3-year-old girl with complete lack of adenosine deaminase (ADA) activity who presented with severe atopic dermatitis and insulin-dependent diabetes mellitus but only mild recurrent infections. Abnormalities of immune function included profound depletion of CD8+ lymphocytes, hyperimmunoglobulinaemia E, and very low in vitro proliferative response to mitogens. Treatment with polyethylene glycol-conjugated ADA was followed by rapid amelioration of clinical and immunological conditions. The immunological and clinical features of this child suggest that the clinical spectrum of ADA deficiency may be broader than originally supposed.