肿瘤坏死因子联合环磷酰胺对H22肝癌小鼠的抗肿瘤作用

肿瘤坏死因子（ＴｕｍｏｒＮｅｃｒｏｓｉｓＦａｃｔｏｒ，ＴＮＦ）和环磷酰胺（Ｃｙｃｌｏ－ｐｈｏｓｐｈａｍｉｄｅ，ＣＹ）联合应用对Ｈ２２肝癌小鼠具有显著协同抗肿瘤作用，这种协同作用与二者用药的先后次序有显著关系。ＴＮＦ、ＣＹ单独应用及ＣＹ先于ＴＮＦ应用的抗瘤作用差（抑瘤率２５．６０％，ＩＬＳ３２．６４％），ＴＮＦ与ＣＹ同时联合应用比单独应用的抗瘤作用略增强（抑瘤率３４．７２％，ＩＬＳ５４．７４％），但如先用ＴＮＦ后用ＣＹ抗瘤作用则显著增强，可以明显改善Ｈ２２肝癌小鼠的免疫功能（ＮＫ活性增长率４９．７０％，ＬＡＫ活性增长率４６．３０％），明显抑制实体瘤生长（抑瘤率６０．１２％），显著延长Ｈ２２肝癌小鼠（腹水型）生存期（ＩＬＳ１０４．７８％），并且有２５％Ｈ２２肝癌小鼠完全治愈。提示：联合ＴＮＦ与ＣＹ治疗肿瘤，ＴＮＦ、ＣＹ序贯用药方法为最佳方案。     

喉癌患者血清可溶性白细胞介素2受体和肿瘤坏死因子的检测及意义

采用双抗体夹心ＥＬＩＳＡ方法，对正常人、声带息肉病人及喉鳞状细胞癌病人的血清可溶性白细胞介素２受体（ｓＩＬ—２Ｒ）和肿瘤坏死因子（ＴＮＦ）水平进行检测。结果表明，喉癌患者血清ＳＩＬ－２Ｒ水平和ＴＮＦ活性均明显高于正常人及声带息肉病人，差异非常显著（Ｐ＜０．０１，Ｐ＜０．００１）。且血清ｓＩＬ－２Ｒ水平与喉癌的病理分化程度、临床分型、分期及手术前后具有明显的相关趋势，ＴＮＦ则不明显。提示：动态观察血清ｓＩＬ－２Ｒ及ＴＮＦ的变化，可作为分析体内抗肿瘤效应、评价疗效及监测预后的参考指标。     

肺癌患者血清sIL—2R和TNF—α的检测及其临床意义

分析了７３例原发性肺癌患者治疗前后血清ｓＩＬ－２Ｒ和ＴＮＦ－α的变化。结果表明：９１）ｓＩＬ－２Ｒ和ＴＮＦ－α均显著高于正常对照组，肿瘤越晚期升高越明显；（２）治疗后ｓＩＬ－２Ｒ和ＴＮＦ－α都明显下降，但各治疗下降程序不同，放、化疗组比手术切除下降更显著；（３）ｓＩＬ－２Ｒ和ＴＮＦ－α在肺癌病人呈正相关机理不明。     

肺癌患者血清sIL－2R和TNF－α的检测及其临床意义

分析了７３例原发性肺癌患者治疗前后（手术切除１６例，放疗２８例，化疗２９例）血清ｓＩＬ－２Ｒ和ＴＮＦ－α的变化。结果表明：（１）ｓＩＬ－２Ｒ和ＴＮＦ－α均显著高于正常对照组（Ｐ＜０．０１），肿瘤越晚期升高越明显（Ｐ＜０．００１）；（２）治疗后ｓＩＬ－２Ｒ和ＴＮＦ－α都明显下降（Ｐ＜０．０１）．但各治疗组下降程度不同，放、化疗组比手术切除组下降更显著；（３）ｓＩＬ－２Ｒ和ＴＮＦ－α在肺癌病人呈正相关（ｒ＝０．３４），机理不明。结果提示，血清ｓＩＬ－２Ｒ和ＴＮＦ－α可作为肺癌患者免疫功能的监测指标之一，并可作为疗效判断的参考。     

人原发性肝癌基因的实验研究───肿瘤坏死因子基因的初步应用

将含人肿瘤坏死因子（ＴＮＦα）ｃＤＮＡ的不同逆转录病毒载体（ＬＮＳ－ｔｎｆα、Ｌ－ｔｎｆαＳＮ及ＬＮＣ－ｔｎｆα）和质粒型真核细胞载体（ｐＳＶＫ３－ｔｎｆα），分别用磷酸钙法导入肝癌细胞ＳＭＭＣ－７７２１中，观察ＴＮＦ的表达水平。结果：重组质粒型表达载体ｐＳＶＫ３－ｔｎｆα表达好，２４小时开始分泌ＴＮＦα（５０Ｕ／ｍｌ），４８～７２小时达最高值（９０Ｕ／ｍｌ），９６小时后下降（４０Ｕ／ｍｌ）。将ｐＳＶＫ３－ｔｎｆα重组ＤＮＡ用磷酸钙包裹后直接进行瘤内注射，发现裸鼠外周血ＴＮＦα水平有一过性增高，３周后测量瘤体大小，发现实验组的肿瘤体积明显小于对照组（２．５±１．６ｃｍ比３．２±１．８ｃｍ，ｎ＝６，Ｐ＜０．０５），实验组荷瘤裸鼠的生存时间亦明显延长（４４．０±３．３ｄ比２８．２±２．０ｄ，ｎ＝６，Ｐ＜０．０１）。结果表明，应用ＴＮＦα基因转移进行肝癌的基因治疗是一条值得探索的途径。     

肿瘤坏死因子与白细胞介素2协同抑制肺腺癌细胞的研究

目的：检测肿瘤坏死因子α（ＴＮＦα）、白细胞介素２（ＩＬ－２）单用或合用对肺腺癌Ａ５４９细胞系（Ａ５４９）和耐顺铂Ａ５４９细胞系（Ａ５４９／ＤＤＰ）的细胞毒性作用。方法：采用四氮唑蓝快速比色法（ＭＴＴ法）进行体外细胞毒性实验。结果：ＴＮＦα对两株细胞的毒性作用无显著差异;与ＩＬ－２合用后,ＴＮＦα对两株细胞的毒性作用分别较单用时均明显增强（Ｐ〈０．０１）。结论：ＴＮＦα与ＩＬ－２对两株细胞具有协同     

肿瘤病人若干免疫功能的观察

用酶联法、同位素掺入法、同位素释放法等检测肿瘤病人血清ｓＩＬ－２Ｒ、ＴＮＦ、ＮＫ细胞活性、ＬＡＫ细胞活性、淋巴细胞转化及淋巴细胞对ＩＬ－２的反应性等免疫功能。结果表明：肿瘤病人血清ｓＩＬ－２Ｒ（２４１．９±１７２．５ｐｍｏｌ／Ｌ）及ＴＮＦ（１．８６±１．５２ｎｇ／ｍｌ）含量均显著高于健康对照组（分别为：１３４．２±７６．５ｐｍｏｌ／Ｌ，Ｐ＜０．０５～０．００５，０．６３±０．２０ｎｇ／ｍｌ）；而肿瘤病人ＮＫ细胞活性（１１．１８±６．９８）、ＬＡＫ细胞活性（１７．８６±９．５７）、淋巴细胞转化（３２．８３±５２．５９）及淋巴细胞对ＩＬ－２的反应性（５．９４±９．３１）等均显著低于健康对照组（分别为：２０．３３±１０．９７，３８．４９±８．１０，７５．７０±５２．６５和２４．５９±２８．２５，Ｐ＜０．０４～０．００１）。结果提示：肿瘤病人存在着多种免疫功能的改变，故应给予合理的过继免疫治疗。     

急性白血病患者外周血几种细胞因子水平的测定及其意义

目的 了解ＩＬ－２等细胞因子在白血病发病机制中的作用。方法 测定了５１例急性白血病患者ＰＢＭＮＣ培养上清中ＩＬ－２,ＩＬ－２,ＩＬ－６水平及血清ｓＩＬ－２Ｒ,ＴＮＦα水平。结果 （１）急性白血病患者ＰＢＭＮＣ培养上清的ＩＬ－２水平显著低于正常（Ｐ〈０．０１）及ＣＲ组（Ｐ〈０．０５）;而ＩＬ－６水平显著高于正常及ＣＲ组（Ｐ〈０．０１）。（２）急性白血病患者血清ｓＩＬ－２Ｒ及ＴＮＦα水平显著高于正常及     

胃癌患者治疗前后血清可溶性肿瘤坏死因子受体Ⅰ的变化及其意义

目的探讨胃癌患者治疗前后血清可溶性坏死因于受体（ｓＴＮＦ－Ⅰ）水平的临床意义。方法应用酶联免疫吸附测定（ＥＬＩＳＡ）的方法测定了３４例胃癌患者ｓＴＮＦＲ－Ⅰ水平。结果胃癌患者血清ｓＴＮＦＲ－Ⅰ水平明显高于正常人（Ｐ＜０．０１），且与临床分期有关，病情越晚，ｓＴＮＦＲ－Ⅰ水平越高，ｓＴＮＦＲ－Ⅰ水平超过３．００ｎｇ／ｍｌ者预后不良；治疗有效者ｓＴＮＦＲ－Ⅰ均显著下降（Ｐ＜０．０１），２４例治疗后４～８个月再次采血检验，８例无瘤生存者ｓＴＮＦＲ－Ⅰ水平降至正常，其余１６例肿瘤未控制或复发转移者血清ｓＴＮＦＲ－Ⅰ水平升高。结论血清ｓＴＮＦＲ－Ⅰ测定对胃癌疗效观察、病情监测及预后判断有重要意义。     

急性白血病患者外周血几种细胞因子水平的测定及其意义

目的了解ＩＬ－２等细胞因子在白血病发病机制中的作用。方法测定了５１例急性白血病患者ＰＢＭＮＣ培养上清中ＩＬ－２、ＩＬ－６水平及血清ｓＩＬ－２Ｒ、ＴＮＦα水平。结果（１）急性白血病患者ＰＢＭＮＣ培养上清的ＩＬ－２水平显著低于正常（Ｐ＜０．０１）及ＣＲ组（Ｐ＜０．０５）;而ＩＬ－６水平显著高于正常及ＣＲ组（Ｐ＜０．０１）。（２）急性白血病患者血清ｓＩＬ－２Ｒ及ＴＮＦα水平显著高于正常及ＣＲ组（Ｐ＜０．０１）。（３）急性白血病患者血清ｓＩＬ－２Ｒ水平与其外周血幼稚细胞比例呈正相关（ｒ＝０．４１０,Ｐ＜０．０５）;在Ｍ５患者中血清ｓＩＬ－２Ｒ与ＴＮＦα水平正相关（ｒ＝０．８７４,Ｐ＜０．０１）。结论急性白血病患者存在着细胞因子网络的失调,并在急性白血病的发生、发展中起一定的作用。     

Comparison of methods for DNA extraction from paraffin-embedded tissues and buccal cells

Both paraffin-embedded tissue specimens and buccal cells are excellent resources for large-scale molecular epidemiological studies. In order to identify the optimal method for DNA extraction, we compared three methods: (1) modified phenol-chloroform protocol; (2) simple boiling method; and (3) DNA Extraction Mini Kit. For paraffin-embedded tissue specimens, amplification of the beta-globin gene sequence was successful in 30 of 34 (88.2%) by the simple boiling method, 29 of 34 (85.3%) samples using DNA extracted by the phenol-chloroform method, and 18 of 34 (52.9%) by the DNA Mini Kit. For buccal cells, amplification of the beta-globin gene sequence was successful in 16 of 17 (94.1%) DNA samples extracted by the phenol-chloroform method, 2 of 16 (12.5%) by the simple boiling method, and 12 of 16 (75%) by the DNA Mini Kit. Both the simple boiling method and the phenol-chloroform method are better methods for DNA isolation from paraffin-embedded tissue specimens, and the phenol-chloroform method is the best method for DNA extraction from buccal cells.     

三氧化二砷对人大肠癌细胞生长及其端粒酶活性的影响

目的探讨三氧化二砷(As2O3)注射液对人类大肠癌细胞生长及其端粒酶活性的影响。方法采用活细胞观察法、台盘兰拒染法和四甲基偶氮唑蓝比色(MTT)法,严密观察As2O3注射液对人大肠癌细胞株LoVo和CoLo-320生长的影响;应用端粒重复序列扩增法(TRAP)—聚丙烯酰胺凝胶电泳(PAGE)银染法检测As2O3注射液处理前、后大肠癌细胞端粒酶活性的变化。结果As2O3注射液能够显著地抑制LoVo和CoLo-320细胞株的生长,并对癌细胞的端粒酶活性具有明显的抑制作用,该作用呈现出一定的浓度和时间依赖性。结论As2O3注射液的确对于人大肠癌细胞株的生长具有显著的抑制作用,该作用可能与As2O3注射液能够抑制癌细胞的端粒酶活性密切相关。     

吉非替尼不同加药时间肺癌A549细胞放射增敏研究

目的 探讨吉非替尼不同加药时间对离体培养肺癌细胞系A549的放射增敏效应的影响.方法 实验分吉非替尼照前24 h加药组、照后即刻加药组、照后24 h加药组.用克隆形成法检测细胞存活分数,流式细胞仪检测细胞凋亡和细胞周期分布,蛋白免疫印迹检测p21、Cdc25e、Bcl-2、Bax、Rad51和phospho-DNA-PK蛋白表达.结果 吉非替尼照前组、照射即刻加药和照后加药组的放射增敏比(D_0值比)分别为2.23、1.51和1.30.照前加药组观察到更高的凋亡率和G_2+M期阻滞.p21、Cdc25c、Bcl-2、Bax、Rad51和phospho-DNA-PK蛋白表达的改变也较其他组明显.结论 吉非替尼照前加药、照射即刻加药和照后加药对肺癌细胞系A549均有放射增敏效应,但照前加药的效应最强.     

FCLA chemiluminescence from sonodynamic action in vitro and in vivo

In this work, the chemiluminescence method was engaged for the first time to detect the active oxygen species during sonodynamic action in vitro and in vivo. We used FCLA (3,7-dihydro-6-[4-[2-[N'-(5-fluoresceinyl)thioureido]-ethoxy]phenyl]-2-methylimidazo[1,2-a]pyrazin-3-one sodium salt), which can selectively react with singlet oxygen ((1)O(2)) or superoxide anion (O(2)(-)) to emit photons, to detect in real time oxygen free radical formation in the sonosensitization of hematoporphyrin derivative. The results show that (1)O(2) is involved in the sonosensitization. In in vivo experiments, a tumor-imaging method by sonodynamic chemiluminescence detection was established. This method could have potential applications in clinics for tumor diagnosis.     

Potentialities of passive thermoacoustic tomography in hyperthermia.

The potentiality of passive thermoacoustic tomography (PTT) is considered theoretically. The problem is formulated to reconstruct the distribution of the thermodynamic temperature at a depth from the acoustic radiation generated by thermal noise and measured by a set of piezotransducers on the body surface. The 2-D inverse problem has been studied. Three mathematical methods of reconstruction were investigated: (1) the least squares method; (2) Tikhonov's regularization method; and (3) the method of elimination of non-physical solutions. The reconstruction in square as well as in rectangular areas was studied. If one uses the least squares method the square area can be separated into 5 x 5 subareas, the typical dimension of every one is 0.5-2 cm for measurements conducted in the frequency range 2-0.5 MHz. The precision deltaT in temperature reconstruction in different layers is about 0.1-0.2 K for smooth distributions of the thermodynamic temperature and 0.1-0.4 K for abrupt distributions, if the number of different scans is about 10(2) and the time of data collecting is about 1 min.     

纤维支气管镜抽吸物中检测HnRNP A2/B1在肺癌诊断中的价值

目的:研究纤维支气管镜(简称纤支镜)抽吸物中异质性细胞核核糖蛋白A2/B1(HnRNP A2/B1)的表达与肺癌的相关性及其用于肺癌诊断的可行性。方法:利用免疫细胞化学的方法检测经病理学或(和)细胞学确诊的肺癌患者和非肺癌患者的纤支镜抽吸物中HnRNP A2/B1的表达情况,并分析其表达水平的差异性。结果:1) HnRNP A2/B1 在肺癌组和非肺癌组患者纤支镜抽吸物中表达阳性率分别为68. 6%(83/121)和13. 6%(3/22),两组比较差异有统计学意义,P<0. 05。2) HnRNPA2/B1在不同病理类型中表达阳性率也不同,鳞癌为67. 6% ( 50/74 ), 腺癌为60. 0%(18/30),小细胞癌为88 2%(15/17),差异有统计学意义,P< 0 .05。结论: HnRNP A2/B1 在肺癌患者纤支镜抽吸物中表达的敏感性为68 .6%(83/121),特异性为86. 4%(19/22),可用于肺癌的诊断。     

Bhas 42细胞转化试验高通量检测方法的建立及应用

目的: 建立Bhas 42细胞转化试验高通量检测方法,并应用于染料木黄酮(GEN)潜在致癌性的检测中。方法:建立Bhas 42细胞转化试验方法,比较细胞灶法和H2O2法对结果判定的影响。细胞灶法试验组在第21天直接固定染色。H2O2法试验组在细胞接种后第19天采用H2O2处理,染色并测定D(450)以确定细胞转化率。计算转化率以验证两种方法的相关性。并通过细胞生长试验选择适宜的GEN浓度进行细胞转化试验,H2O2法判定转化试验结果。结果:在细胞灶法中,启动试验3-甲基胆蒽(3-MCA)组、促癌试验佛波醇酯(TPA)组的转化灶个数分别与启动、促癌试验DMSO组相比明显升高(P2O2法中,启动试验3-MCA组、促癌试验TPA组的转化灶个数、D(450)分别与启动、促癌试验DMSO组相比均明显升高(PD(450)相对于阴性组有显著差异(P结论:成功建立了Bhas 42细胞转化试验的H2O2法并实现了高通量检测,该法进一步缩短了实验周期并提高了结果的客观性,适用于非遗传毒性致癌物的体外早期筛选。GEN在促癌试验中呈阳性,但在启动试验中呈阴性,提示其可能是非遗传毒性致癌物。     

雄黄诱导白血病细胞凋亡的研究

目的探讨雄黄对白血病细胞凋亡及Bc l-2表达水平的影响。方法以阿霉素作用组为阳性对照组,通过四甲基偶氮唑盐法(MTT)测定药物作用下细胞株的细胞活力(OD值),采用原位细胞凋亡检测法(TUNEL)观察凋亡细胞的形态学改变,用流式细胞仪法(FCM)测定药物作用下细胞株Bc l-2蛋白阳性表达率的变化。结果与ADR相比,雄黄作用组OD值和Bc l-2阳性表达率更低(P<0.05),凋亡率更高(P<0.05)。结论雄黄诱导白血病细胞凋亡的效果较好,且Bc l-2蛋白表达水平的变化能显著改变药物诱导细胞凋亡的能力。     

Effectiveness of 3 ways method in intra-arterial infusion chemotherapy for advanced or recurrent cervical cancer with or without radiotherapy

We analyzed 58 cases of advanced or recurrent cervical cancer treated with intra-arterial infusion chemotherapy (IAIC) with or without radiotherapy. Two separate IAIC regimens were administered since 1985: group I consisted of 5-FU + MMC +/- ADR (30 patients) and group II consisted of CDDP + MMC +/- 5-FU (28 patients). The tip of a catheter was placed in the bifurcation of the abdominal aorta (1 way method: 45 patients regimen II: 15) between 1977-1984. We have used selective catheterization (2 ways method: 9) since 1995 in order to get good drug distribution. However we experienced grade 4 toxic effect of cutaneous and pain with this method, so we have used a 3 ways method (4 patients) since 1998. The two-year survival rete was 60% with the 1 way method, and 67% with the 2 ways method and regimen II. Severe adverse effects (grade 3 + 4) were found in 53, 56, 0%, respectively, by each of the three methods (1, 2, 3 ways) hematologically, 13, 22, 0% in gastrointestinally, 0, 44, 0% in cutaneously and 0, 56, 0% in pain or neurotoxicity. These data suggest that IAIC by the 3 ways method is a useful treatment for advanced or recurrent cervical cancer. However, one should check the blood flow distribution periodically, and control concentration of drugs.     

Localization of tumor by chemiluminescence probe during photosensitization action

As a chemiluminescence (CL) probe, 3,7-dihydro-6-[4-[2-(N'-(5-fluoresceinyl) thioureido)ethoxy]phenyl]-2-methylimidazo[1,2-a]pyrazin-3-one dosium salt (FCLA) can sensitively and specifically react with singlet oxygen ((1)O(2)) and superoxide (O(2)(-)). Based on the fact that photosensitization action involves (1)O(2) generation, this report presents a novel method for localization of tumor mediated by FCLA. In experiments, FCLA was used as an optical reporter of (1)O(2) produced from photosensitization reaction of hematoporphyrin derivative. The effects of azide inhibition and deuterium enhancement on photosensitized CL indicated that (1)O(2) molecules generated during photosensitization action could be measured and imaged. In vivo experiments in vivo with tumor-bearing nude mice showed a clear CL image of tumor. The study suggests that CL imaging method may be applicable to the detection of (1)O(2) and the diagnosis of superficial tumors.