结直肠癌患者肿瘤组织与外周血中MTHFR C677T的多态性及相关性研究

目的 探索外周血代替肿瘤组织进行MTHFR C677T基因多态性检测的可行性.方法 选取首诊为结直肠癌且行根治性手术的患者26例,术前未接受任何化学治疗,应用聚合酶链反应-限制性内切片段长度多态性（PCR-RFLP）方法检测MTHFR C677T的基因多态性,比较外周血及肿瘤组织中相关基因型的一致性.结果 26例结直肠癌患者的肿瘤组织及外周血中MTHFR 677CT基因型、TT基因型、CC基因型一致的病例数分别为7、7、4例.肿瘤组织和外周血中MTHFR C677T的基因型一致率为69.2%.结论 外周血中MTHFR C677T的基因多态性可以反映肿瘤组织中相关基因型的突变.     

DNA arrays to monitor gene expression in rat blood and uterus following 17beta-estradiol exposure: biomonitoring environmental effects using surrogate tissues.

We propose that gene expression changes in accessible tissues such as blood often reflect those in inaccessible tissues, thus offering a convenient biomonitoring method to provide insight into the effects of environmental toxicants on such tissues. In this pilot study, gene expression changes in peripheral blood leukocytes (PBL) were compared to those in the uteri of adult rats to identify genes that were altered in both tissues following estradiol treatment. Ovariectomized rats were treated with either 17beta-estradiol or vehicle control (corn oil) for 3 days. PBL and uterine RNAs were hybridized to arrays containing 1185 genes. One hundred and ninety three genes were expressed in common between the PBL and uterus. Eighteen were changed significantly in both tissues, 9 of which were treatment- but not tissue-specific (e.g., jun-D, phospholipase A2, thymidine kinase). These results demonstrate that many genes are coexpressed between PBL and uterus, and that some are coregulated by estradiol. Given the limited number of genes examined in this study and the estimated size of other mammalian genomes, we conclude that many more genes will also be coregulated and suggest that accessible tissues such as PBL can serve as surrogate tissues for observing gene expression changes in inaccessible target tissues.     

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。     

不同采血方法在血常规相关检验中的应用价值对比分析

目的探讨不同采血方法在血常规相关检验中的应用价值。方法笔者所在医院进行体检的健康人员90例,随机分为静脉组和末梢组各45例。分别采集静脉血和末梢血进行血常规检查,比较两组人员血常规检查中的白细胞计数、红细胞计数、红细胞比容、血红蛋白、平均红细胞血红蛋白浓度以及血小板等检查项目。结果静脉组45例患者的血常规检查结果为,WBC:(5.24±3.16)×109/L;RBC:(3.34±0.98)×1012/L;HCT:(42.35±20.46)%;HGB:(115.75±15.76)g/L;MCHC:(319.78±28.79)g/L;PLT:(228.75±84.78)×109/L。末梢组45例患者的血常规检查结果为,WBC:(5.86±3.18)×109/L;RBC:(3.86±0.87)×1012/L;HCT:(31.46±21.48)%;HGB:(101.56±16.48)g/L;MCHC:(322.43±28.94)g/L;PLT:(213.48±81.46)×109/L。两组患者的MCHC检查结果相比较,差异无统计学意义(P>0.05),而WBC、RBC、HCT、HGB、PLT的检查结果相比较,差异均有统计学意义(P<0.05)。结论不同的采血方法对血常规的检验结果具有很大的影响,应用静脉血代替末梢血可以有效的提高检验结果的准确性、提高仪器设备的使用寿命,值得在临床上推广使用。     

Transdermal toxicity of topically applied anticoagulant rodenticide warfarin in rats

Occupational/accidental exposure data have showed hemorrhage as a result of transdermal exposure to warfarin, however, other effects are not known. In the present study, the impact of epicutaneous application of 10 μg or 100 μg of warfarin (three times, once a day) on peripheral blood polymorphonuclear (PMN) and mononuclear cells (PBMC) was examined in rats. Both doses resulted in prolongation of prothrombin time and changes in hematologic parameters. Increases in PMN intracellular myeloperoxidase (MPO) activity were seen at higher warfarin dose and both doses resulted in higher percentages of granular CD11b⁺ cells. In contrast, a decrease in PMN TNF and IL-6 production (ELISA) and gene expression (RT-PCR) was observed. Epicutaneous application of warfarin resulted in decreased numbers of PBMC, higher numbers of mononuclear CD11b⁺ cells, but without effect on PMBC cytokine production. The data obtained showed differential effects of transdermal exposure to warfarin depending on leukocyte type and activity.     

胃癌患者外周血中端粒酶活性表达及其临床意义

目的 检测胃癌患外周血中癌细胞端粒酶活性，以探讨端粒酶活性的表达及其临床意义。方法 通过用PCR-TRAP-ELISA法检测52例胃癌患外周血端粒酶的活性，按照肿瘤大小、分化程度、浸润深度、淋巴结有无转移来分析端粒酶与胃癌的关系。结果 外周血中检测端粒酶活性阳性率为53．8％，低于相应的胃癌组织阳性率(81．3％)，胃癌患外周血中肿瘤细胞端粒酶相对活性的升高与患的淋巴结转移和肿瘤的分化程度呈正相关。结论 对患判断肿瘤是否转移复发，检测患外周血中端粒酶活性要优于检测肿瘤组织的端粒酶活性。     

末梢血与静脉血检测糖化血红蛋白结果的对比分析

目的 探讨用末梢血与静脉血检测糖化血红蛋白(HbA1c)结果的可比性.方法 313例研究对象同时取末梢血与静脉血用Primus PDQ PLUS糖化血红蛋白分析仪检测HbA1c,对检测结果进行相关性分析和配对t检验.结果 用末梢血与静脉血检测HbA1c的结果数据经统计学分析有显著相关性(健康组、糖尿病组、全体分别r=0.97、0.98、0.99,均P＜0.01),差异无统计学意义(健康组、糖尿病组、全体分别P=-0.77,P=0.72,P=0.75).结论 可以用末梢血检测HbA1c,其结果与静脉血检测HbA1c的结果无明显差异,可作为糖尿病患者的血糖检测指标.     

Hematological cell findings in bone marrow and peripheral blood of rabbits after experimental acute exposure to Loxosceles intermedia (brown spider) venom.

The purpose of this work was to find out the cellular changes occurring in bone marrow and peripheral blood after acute exposure to the venom of Loxosceles intermedia. Doses of 40 microg of venom were injected intradermally into five rabbits, and five rabbits receiving only phosphate-buffered saline (PBS) were used as controls. Bone marrow and peripheral blood samples were obtained before the envenomation and 4, 8, 12, 24 and 48 h, and 5, 10, 15, 20 and 30 days after envenomation. In bone marrow samples we assessed cellularity, nucleated red cells, megakaryocytes and neutrophils, and in peripheral blood we assessed red cells (red cell concentration, hemoglobin and hematocrit), leukocytes, neutrophils and platelets. Our objective was to find out if the venom has a direct effect on bone marrow and peripheral blood or if changes in both of them are secondary to the needs of tissues, and if there is a good correlation between histopathological and hematological findings. We found that the red cell parameters were not affected by the venom, except for nucleated red cells which decreased after venom exposure. The depression of megakaryocyte numbers and thrombocytopenia showed a strong correlation with the histopathologic changes observed in skin biopsies obtained from the rabbits. The changes in cellularity and neutrophils of bone marrow were strongly correlated with those in peripheral blood and skin. The thrombocytopenia and neutropenia in peripheral blood are due to marrow depression, which may be a consequence of an extensive migration of platelets and neutrophils to the necrotic lesion or the marrow depression may be a transitory effect of evenoming by L. intermedia.     

Analysis of cytogenetic damage in rat lymphocytes following in vivo exposure to nitrobenzene

Male F-344 rats were exposed to target concentrations of 0, 5, 16, or 50 ppm nitrobenzene for 6 h/day, 5 days a week, for 21 days during a 29-day period. Isolated spleen (ISLs) and peripheral blood lymphocytes (PBLs) were cultured in the presence of 2 microM 5-bromodeoxyuridine and scored for sister-chromatid exchange (SCE) and PBLs were also scored for chromosome aberrations. No significant increase in SCE frequency or chromosome aberrations was found in the PBLs, and no significant increase in SCEs was observed in the ISLs at any of the concentrations. Thus, cytogenetic analysis of ISLs and PBLs provide no evidence of a genotoxic potential for nitrobenzene.     

Genomic and genetic biomarkers of toxicity

Biomarkers in general use today are diagnostic in that they reflect an organ's ongoing dysfunction or damage. Unfortunately, in some cases the change in the biomarker occurs too late for effective medical intervention such as seen with acute renal failure. New biomarkers of toxicity are needed to (a) alert physicians of subtle changes prior to organ dysfunction or damage to enable preventive measures and (b) predict, prior to human exposure, if a drug is likely to induce toxicity in many patients or in specific individuals. Microarray technologies can move biomarker discovery forward at an unforeseen speed as tens of thousands of genes and genetic variants can be monitored simultaneously in one biological specimen. Pharmacogenomics, herein defined as the study of a drug's effect on gene expression, can be used to discover biomarkers in solid tissues or peripheral blood cells that are altered in animals or individuals following drug exposure. Pharmacogenetics, herein defined as the study of genetic factors that affect drug response, can be employed to identify individuals whose genetic make-up suggests they would respond poorly to a particular drug. Biomarkers discovered with these approaches can result in genomic/genetic tests, protein assays or other analytical tests. Examples of such are provided with a discussion of the unresolved issues that inhibit the use of toxicity biomarkers such as biomarker validation, reimbursement of clinical tests and patient privacy.     

Blood content in guinea-pig tissues: Correction for the study of drug tissue distribution

The blood content in different organs or tissues of guinea-pigs was determined by means of [125I]bovine serum albumin. The blood distribution expressed as percentage of total body blood for various organs or tissues ranged from 0.35 (adipose) to 17.5 (muscle)%. The blood content in tissue preparations expressed as the blood background correction factor, F (ml blood/g wet tissue) depended on the extent of bleeding during experiment; that factor can cause a considerable difference in the results of a tissue distribution study of a drug. Blood content was high in the lung (0.36 ml/g), heart, liver, kidney and spleen, but low in adipose and brain tissues (0.021 ml/g). The distribution of valproic acid in various tissues of guinea-pigs after intravenous injection was determined from the homogenates of isolated organs. The results showed that blood contamination can greatly alter the data of tissue distribution of a drug to as much as 25%. This study proposed a maximum (non-bled) and a minimum (extreme-bled) blood background correction factor for respective tissues of guinea-pigs, as well as equations for correcting the blood contamination in tissue during the drug distribution study. The results suggest that to obtain an exact analysis of drug concentrations in specific tissues, a correction for the blood of that specific tissue is necessary.     

糖皮质激素对哮喘大鼠血单个核细胞及肺组织ICAM-1表达的影响

目的　探讨细胞间粘附分子 1(ICAM 1)在哮喘气道炎症粘附机制中的作用及糖皮质激素对哮喘大鼠血单个核细胞及肺组织ICAM 1表达的影响。方法　将SD大鼠随机分成哮喘组、地塞米松治疗组和正常对照组 ,采用免疫细胞化学和免疫组织化学研究各组大鼠血单个核细胞和肺组织ICAM 1表达情况。结果　 (1)ICAM 1主要表达在肺小血管、毛细血管内皮细胞、支气管、肺泡上皮细胞、粘膜下基底膜及浸润的炎细胞上 ,哮喘组大鼠肺组织ICAM 1表达显著高于地塞米松治疗组和正常对照组 (P <0 0 5 )。 (2 )地塞米松组大鼠单个核细胞ICAM 1表达较哮喘组减轻 (P <0 0 5 )。 (3)治疗组大鼠肺组织和气管炎症反应以及气道反应性较哮喘组减轻。结论　证实了I CAM 1在哮喘炎性反应中的作用 ,地塞米松抑制ICAM 1表达。     

Topoisomerase II beta expression level correlates with doxorubicin-induced apoptosis in peripheral blood cells

Anthracyclines are widely used in oncology. Both the response and side-effects of anthracyclines are individually variable, but determinants or predictive markers of this variability are not available. We investigated the variability in the expression of the anthracycline targets topoisomerases II (topo II) alpha and beta and its significance for the apoptotic response following exposure to the anthracycline doxorubicin. Only topo II beta protein expression was detected in peripheral blood cells. Usually considered a constitutively expressed protein, topo II beta varied 3-, 18-, and 16-fold on the mRNA, protein and activity levels, respectively, among the volunteers tested. In addition, the expression of topo II beta was modified by several mitogens, suggesting a role in the regulation of cell cycle. Strikingly, topo II beta activity correlated statistically significantly with the apoptotic response in peripheral blood leukocytes exposed to 1 μM doxorubicin. A longitudinal study in a subset of study subjects demonstrated that 30% of the topo II expression variability may be inherited. However, resequencing of the TOP2B gene in 48 unrelated individuals revealed only 8 gene variants, none of them with obvious effects on the expression or protein sequence of topo II beta. Taken together, the apoptotic response to doxorubicin in peripheral blood cells may be mediated by topo II beta. The expression level of topo II beta is intra- and inter-individually variable, and may in part determine the apoptotic response to doxorubicin and other anthracyclines.     

黄芩苷对牙周炎大鼠牙龈组织及外周血中 IL-8表达的影响

目的：采用大肠杆菌脂多糖诱导大鼠牙周炎模型,观察黄芩苷对牙周炎大鼠牙龈组织及外周血中白介素-8（IL-8）表达水平的影响。方法选用50只8周龄SD 大鼠,随机分成5组,每组10只,A组为正常对照组,B组为牙周炎组,C1、C2、C3组为黄芩苷治疗组（分别以0．01、0．1、1．0μg／ml黄芩苷治疗）,治疗1周后处死大鼠采取其外周血及牙龈组织,采用微量样本多重蛋白定量技术（ CBA）检测外周血中IL-8表达含量,采用酶联免疫法（ ELISA）检测牙龈组织中IL-8的表达含量。结果牙周炎组大鼠牙龈组织及外周血中IL-8表达含量均明显高于正常对照组（ P ＜0．05）,黄芩苷治疗组大鼠牙龈组织中IL-8和外周血中IL-8表达含量均明显低于牙周炎组（ P ＜0．05）,且C1、C2、C3组IL-8水平逐渐下降（ P ＜0．05）。结论黄芩苷可以降低牙周炎大鼠牙龈组织和外周血中IL-8的表达含量,抑制脂多糖对大鼠牙周组织的损伤作用。     

Sperm evaluation in human genetic monitoring

Health hazards, including reproductive hazards, resulting from exposure to mutagenic chemical or physical agents are a major concern. Human genetic monitoring is a relatively new approach to the evaluation of human population exposure to mutagens that is based on the use of short-term tests with human cells (including sperm), tissues, and body fluids for genetic damage. Evaluation of sperm for count, percentage of morphologically abnormal forms, and frequency of heads containing zero, one, or two fluorescent bodies have been incorporated in genetic monitoring studies. In this paper, the application of human genetic monitoring to the evaluation of germ cell mutagen exposure is discussed. The purposes and limitations of human genetic monitoring studies are presented. Some of the practical issues involved in the use of sperm studies in genetic monitoring are described through the examples of two studies in different settings that were conducted by the author's laboratory. The first was a study of Autopsy Service workers within the resident institution of the laboratory. The second was a study of workers exposed to coal tar pitch volatiles at an industrial plant approximately 1,000 miles from the laboratory.     

Measurement of micronuclei by cytokinesis-block method in human, cattle, goat, pig, rabbit, chicken and fish peripheral blood lymphocytes irradiated in vitro with gamma radiation

The frequencies of gamma-ray-induced micronuclei (MN) in cytokinesis-blocked (CB) lymphocytes at several doses were measured in three donors of seven species (human, cattle, goat, pig, rabbit, chicken, fish). Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors of human, cattle, goat, pig and rabbit. The relative sensitivity of cattle, goat, pig and rabbit in peripheral blood lymphocytes (PBLs) compared with human PBLs was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 cGy to 400 cGy. In the case of MN frequency with 0.2, the relative sensitivities of cattle, goat, pig and rabbit PBLs were 0.86, 0.98, 0.41 and 0.39, respectively. These data indicate that the induction of MN in CB cells following irradiation is similar in human, cattle and goat PBLs, while PBLs from pig and rabbit were much less sensitive to the MN induction effects of gamma-radiation than those from human. The micronucleus counts failed to show any evidence of radiation damage in the cells from chicken and fish. Measurements performed after irradiation showed a dose-related decrease in the formation of binucleated cells. We concluded that the use of CB cell from fish and chicken for detecting the results of radiation exposure was highly questionable. Our in vitro radiobiological study confirmed that the cytogenetic response obtained in blood from selected breeds of mammalian species can be utilized for application in environmental studies.     

痛风性关节炎患者外周血白细胞中Toll样受体4表达与基因多态性研究

目的 对痛风性关节炎患者外周血白细胞中Toll样受体4(TLR4)表达与基因多态性予以分析,为痛风性关节炎的早期诊断和治疗提供参考.方法 选择2015年1月—2016年6月右江民族医学院附属医院确诊为痛风性关节炎患者及同期体检健康者为观察组和对照组,每组各80例.均行外周血白细胞中TLR4表达测定和基因型鉴定并进行比较和评价.结果 观察组外周血白细胞中TLR4 mRNA、核因子-κBD65、血浆IL-1β及血尿酸水平均显著高于对照组(P<0.05);观察组GGb、GT及G等位基因b与对照组比较差异无统计学意义(P>0.05),观察组TT和T等位基因均显著高于对照组(P<0.05).结论 痛风性关节炎患者外周血白细胞中TLR4表达与TT和T等位基因均表现出较强的表达及较高的的比率,对临床诊断及治疗具有一定的指导价值.     

基因芯片筛查外周血中胃癌转移相关基因

目的应用基因芯片筛查外周血中胃癌转移相关基因,为探究外周血中胃癌转移分子标志物提供线索。方法采用phalanx人全基因组表达谱芯片检测胃癌发生转移患者与未发生转移患者外周血中基因表达谱差异,利用生物信息学技术分析检测结果。结果从外周血中筛选出胃癌转移相关差异表达基因157个,表达上调基因92个,表达下调基因65个,涉及细胞粘附和运动基因、蛋白水解酶基因等。结论胃癌转移过程涉及多个不同功能基因表达的改变,运用基因芯片技术可初步从外周血中筛选出胃癌转移相关基因。