脂蛋白残粒RLP-c检测的临床意义

目的 探讨脂蛋白残粒RLP-c作为新的脂质指标的临床意义。方法 采用免疫分离法测定正常对照组(NC)、冠心病组(CHD)和2型糖尿病组(T2DM)的RLP-c水平,并比较RLP-c与其他脂质指标的相关性。结果 CHD组和T2DM组RLP-c水平明显高于NC组(P<0.01),其水平与甘油三脂(TG)和极低密度脂蛋白胆固醇(VLDL-c)高度相关(P<0.01)。结论 RLP-c可用于对动脉粥样硬化的危险性评估。     

马齿苋活性成分体内外抗癌作用的初步筛选

目的 观察马齿苋活性成分对人肺腺癌细胞系(A-549 cell)、人喉表皮样癌细胞系(Hep-2 cell)、人宫颈癌细胞系(Hela cell)和人恶性胚胎横纹肌瘤细胞系(RD cell)生长的影响.方法 体外培养条件下用不同浓度的马齿苋活性成分处理4种癌细胞,通过溴化二甲噻唑二苯四氮唑(MTT)试验法测定癌细胞的增殖;同时处理S180荷瘤小鼠,观察荷瘤小鼠的体重、瘤重、死亡率和抑瘤率.结果 马齿苋生物碱对离体培养的A-549肺癌细胞、Hela细胞和Hep-2细胞的增殖均具有明显抑制作用,马齿苋多糖对Hela细胞有较强的抑制作用, 马齿苋脂肪酸对Hep-2细胞有一定的抑制作用,马齿苋黄酮对RD细胞有很强的抑制作用,并存在浓度剂量效应,高浓度抑制作用强.结论 马齿苋活性成分能选择性地杀伤癌细胞,有进一步研究的潜在价值.     

正常人外周血淋巴细胞 IL—2受体免疫放射测定

采用~(125)I 标记小鼠抗人IL—2受体(P55)的抗Tac(CD_(25))单克隆抗体,成功地建立了人IL—2受体的免疫放射分析法,动态观察了人外周血淋巴细胞经PHA 刺激24、48和72h 后IL—2受体表达的时间曲线,通过Scatchard 作图分析表明~(125)I—抗Tac 单克隆抗体与PHA 活化的上述三个时间点的T 淋巴细胞的最大结合容量(B_(max))分别为43000位点/细胞、54000位点/细胞和61000位点/细胞。本研究为临床IL—2受体检测提供一种简便的免疫放射测定法。     

ReProComet: a new in vitro method to assess DNA damage in mammalian sperm.

The increasing request of chemical safety assessment demands for the validation of alternative methods to reduce the resort to animal experimentation. Methods that evaluate reproductive toxicity are among those requiring the largest use of animals. Presently, no validated in vitro alternative exists for the assessment of reproductive toxicity. Mammalian sperm are sensitive targets of DNA-reactive chemicals, which form premutagenic adducts. Here, we propose a new method based on comet assay to detect DNA damage induced by potential germ cell mutagens in bull sperm available from assisted reproduction practices. In somatic cells, chemical-induced adducts can be revealed by comet assay that detects DNA breaks produced during adduct repair. Mature sperm, however, are devoid of repair enzymes, and adducts are processed only after fertilization. For this reason, comet assay is not sensitive to detect DNA lesions induced in sperm by most chemicals. To overcome such limitation, we developed a modified comet assay based on the addition of a protein extract from HeLa cells to agarose-embedded sperm on microscopic slides. To test the method, sperm were treated in vitro with methyl methanesulfonate (MMS) or melphalan (MLP) and comet assay was conducted both with and without protein supplementation. No effect of MMS or MLP was detected without protein supplementation; on the contrary, a clear-cut dose-dependent effect was measured after addition of the cell extract. These results represent a proof of concept of a novel in vitro mutagenicity test on sperm that could offer a promising approach to complement previously validated in vivo germ cell genotoxicity assays.     

Development of HPLC plasma assays for CAM 4515 and CAM 4750, two new nonpeptide tachykinin antagonists, and application to bioavailability studies

CAM 4515 and CAM 4750 are new nonpeptide tachykinin NK₁ receptor antagonists with different lipophilicities. Two separate, simple, and sensitive HPLC methods for the quantitation of these two compounds in plasma and the evaluation of their oral bioavailability in rats were developed and validated. Extraction of CAM 4515 from plasma involved protein precipitation with acetonitrile, while that for CAM 4750 involved a one-step liquid-liquid extraction with methylene chloride. The analytes in extracts were chromatographed on a C18 column using two different separation buffers, 47% 0.02 M sodium citrate (pH 3.5)-53% acetonitrile for CAM 4515 and 59% 0.02 M potassium phosphate dibasic (pH 7.0)-41% acetonitrile for CAM 4750, and both compounds were detected by fluorescence (excitation 278 nm; emission 342 nm). Stability profiles of both drugs at −20°C or room temperature in plasma and in reconstituted buffers were good. The limit of quantitation for both drugs was 5 ng ml⁻¹ with good linearity from 5 to 1000 ng ml⁻¹ using 100–200 μl of plasma. Excellent precision (relative standard deviation < 8.3%) and accuracy (relative error ± 9.2%) were observed for both CAM 4515 and CAM 4750. Oral bioavailability studies were conducted for each compound in rats receiving a p.o. dose of 20 mg kg⁻¹ and an i.v. dose of 5 mg kg⁻¹. The absolute oral bioavailability of CAM 4750 (80%) was estimated to be 40-fold greater than that of CAM 4515 (2%). The experimental results suggest that incorporation of a pyridine group into the structural backbone may greatly improve bioavailability.     

白细胞介素1α对鼠甲状腺细胞增殖的影响

采用四氮唑蓝（MTT）比色法检测TSH存在和缺乏时白细胞介素1α（IL－1α）对鼠FRTL-5细胞增殖的影响。结果提示，在无TSH条件下IL-1α20～2000kU／L对FRTL－5细胞无明显促增殖作用；在TSH存在时IL－1α20和200kU／L对FRTL－5细胞增殖亦无明显抑制作用，IL－1α2000kU／L则可显著抑制TSH介导的FRTL－5细胞增殖。     

人IL_2活性测定及其影响因素的探讨

作者建立了小鼠胸腺细胞MTT比色分析法并检测了33份正常人的IL_2活性。探讨了影响人IL_2活性测定的因素并与~3H-TdR掺入法进行比较。结果表明,该法简便,稳定,可行,与~3H-TdR掺入法比较二者有较好的一致性,认为本法可用于临床IL_2样本的检测。     

胃癌病人天然杀伤细胞活性监测

报道微量细胞毒试验——LDH 释放法用于对胃癌病人临床免疫的监测.测定100例正常人和50例胃癌病人外周血 NK 细胞毒.结果:正常人值为36.10±5.68,胃癌组中,10例早期胃癌为28.76±4.70,25例进展期胃癌为14.51±5.80,15例晚期胃癌为11.86±4.43,与正常人值比较,P 值分别>0.05,<0.01和<0.01.NK 细胞毒活性随胃癌病变的进展和肿瘤体积的增大而逐渐降低,手术切除肿瘤后可有不同程度回升,甚至恢复到正常水平。长期化疗能损伤 NK 活性.     

Lectin-induced increase in clonogenic growth of haematopoietic progenitor cells

Abstract: The galactoside-specific plant lectin, Viscum album agglutinin (VAA-I) increases cellular parameters of natural host defence. It also binds to a variety of haematopoietic cells, including progenitors. We investigated whether VAA-I has a stimulatory effect on haematopoietic progenitor cells. Peripheral blood progenitor cells from 7 healthy volunteers were cultured in a colony assay with VAA-I plus erythropoietin (EPO) and stem cell factor (SCF). At 50 pg/ml VAA-I induced a significant increase in the cytokine-dependent clonogenic growth (52% in median, p<0.05). In another set of experiments purified CD34+ cells were isolated from the bone marrow aspirate of 4 patients with non-metastatic breast cancer using fluorescence-activated cell sorting. Binding to CD34+ cells was demonstrated by using directly fluorescence-conjugated VAA-I. Co-incubation with d -galactose significantly abrogated this effect. CD34+ cells were cultured in the presence of EPO, SCF, interleukin-3, granulocyte/monocyte colony-stimulating factor and granulocyte colony-stimulating factor. VAA-I alone had no measurable effect on the clonogenic growth of the isolated cells. However, at concentrations of 100 and 250 pg/ml VAA-I increased the cytokine-dependent proliferation and differentiation of CD34+ cells by a median of 75 and 85%, respectively. The results show that VAA-I binds to haematopoietic progenitor cells and has a co-stimulatory effect on their proliferation.     

黄曲霉毒素B_1及其代谢物与原发性肝癌关系的流行病学调查

本文应用酶联免疫测定法对广西壮族自治区原发性肝癌高发的扶绥县和低发的阳朔县,进行了黄曲霉毒素B_1及代谢物黄曲霉毒素M_1和原发性肝癌关系的流行病调查。结果发现,原发性肝癌的死亡率与黄曲霉毒素B_1摄入量,晨尿黄曲霉毒素M_1排出量呈明显的正相关。展尿黄曲霉毒素M_1排出量可作为人群接触黄曲霉毒素B_1水平的指标。原发性肝癌死亡率与玉米、花生和花生油的摄入量有关。