Tactivin疗法对外伤后骨髓病人IL-1β,TNFα的调节

采用Tactivin疗法对5例下肢外伤后骨髓炎病人治疗前后IL-1β，TNFα的变化进行了观察，对照组为健康人。于Tactivin疗法治疗前后采病人血液，分别检测：①血清IL-1β,TNFα水平；②体外培养外周血单个核细胞自发性和经LPS刺激后分泌的IL-IB，TNFα水平（用免疫酶方法检测）；③外周血单个核细胞的1IL-1β，TNFαmRNA基因表型的检测，用N．Gough方法抽提外周血单个核细胞的mRNA，与用’‘P标记的IL－lpDNA探针、TNFaDNA探针分别进行杂交，用放射自显影方法检测。实验结果表明：①来治疗的骨髓病人血清IL－16水平升高的同…     

特发性血小板减少性紫癜患者糖皮质激素受体的表达

目的探讨特发性血小板减少性紫癜(ITP)患者糖皮质激素治疗效应与外周血单个核细胞(PBMC)内两种糖皮质激素受体(GR)亚型-GRα和GRβ,及C-FOX表达水平之间的关系。方法用逆转录-聚合酶链反应(RT-PCR),检测激素敏感型、激素抵抗型ITP患者和正常对照组GR,αGRβ,C-FOX的mRNA的表达。结果ITP患者和正常对照组均有GRα、GRβ,C-FOSmRNA的表达;激素抵抗型ITP患者GRβmRNA表达明显高于激素敏感型ITP患者及正常对照组(P<0.05);GRα,C-FOXmRNA在各组无显著差别。结论ITP患者糖皮质激素抵抗可能与GRβ的表达亢进有关。     

慢性乙型肝炎患者外周血单个核细胞Toll样受体3的表达降低

目的 探讨慢性乙型肝炎患者外周血单个核细胞Toll样受体3(TLR3)的表达及其临床意义.方法 分别采集慢性乙型肝炎患者和健康志愿者外周血,荧光定量PCR法检测血清HBV DNA复制水平;使用RT-PCR、流式细胞术以及免疫印迹技术分别检测外周血单个核细胞TLR3的mRNA、蛋白的表达;使用ELISA法检测血清中肿瘤坏死因子α(TNF-α)和干扰素β(IFN-p)水平.结果 慢性乙型肝炎患者外周血单个核细胞中的TLR3表达显著低于健康志愿者,且降低水平与血清HBV DNA复制水平相关;慢性乙型肝炎患者外周血TNF-α、IFN-β浓度显著低于健康志愿者,且降低的水平与血清HBV DNA复制水平相关.结论 慢性乙型肝炎患者外周血单个核细胞TLR3的表达与乙肝病毒的复制水平相关.     

早期自然流产过程转化生长因子-β1的异常表达

目的 探讨自然流产患者外周血单个核细胞和胎盘组织转化生长因子-β1(TGF-β1)的异常表达.方法 采用逆转录-聚合酶链反应(RT-PCR)技术,半定量检测30例早期自然流产患者、25例正常妊娠妇女、25例正常未妊娠妇女外周血单个核细胞内TGF-β1mRNA表达;采用免疫组织化学技术检测绒毛和蜕膜组织内TGF-β1表达强度.结果 早期自然流产患者、正常妊娠妇女、正常未妊娠妇女3组研究对象外周血单个核细胞表达TGF-β1mRNA相对含量分别为21.5±9.4%、95.7±12.9%、87.0±13.8%,组间比较均有显著性差异(P＜0.05);自然流产组合体滋养层细胞表达TGF-β1显著低于人工流产组(P＜0.01).结论 正常早期妊娠妇女外周血单个核细胞TGF-β1mRNA表达水平较正常未妊娠妇女显著升高,提示TGF-β1对妊娠维护起重要作用;早期自然流产患者外周血单个核细胞TGF-β1 mRNA表达水平及绒毛合体滋养细胞层表达TGF-β1强度均较正常妊娠妇女显著下降,其低水平表达可能与自然流产相关.     

原发性肾病综合征中HSP90与糖皮质激素抵抗关系的研究

目的：观察原发性肾病综合征（INS）患者外周血单个核细胞（PBMC）内热休克蛋白90（HSP90）的表达、分布及其与糖皮质激素受体（GR）之间的关系，以揭示HSP90在INS发病、糖皮质激索抵抗及治疗效应中的作用本质。方法：采用RT-PCR和激光共聚焦免疫荧光技术，分别检测PBMC中HSP90、GR的mRNA表达水平及HSP90的亚细胞分布状况。结果：发现INS患者与健康对照者PBMC中的GR表达无明显差异（P〉0．05）；HSP90的表达较健康对照组明显增高，并且激素抵抗组INS患者HSP90mRNA表达显著高于激素敏感组INS患者（P〈0．05）。INS患者的HSP90／GR比值明显高于健康对照组（P〈0．05）。HSP90亚细胞分布研究发现，健康对照组的HSP90主要分布在PBMC的胞浆中，核内含量极微；INS患者增加的HSP90则明显趋向细胞核内分布（P〈0．001）。结论：HSP90的异常表达及亚细胞分布变化可能是导致内源性、外源性糖皮质激素抵抗的重要机制，其一方面通过影响糖皮质激素对免疫系统的自身反馈调节，参与INS发病；另一方面则可能干预患者对外源性激素的治疗反应，影响临床疗效。     

系统性红斑狼疮病人环孢素A受体mRNA的表达

目的 探讨静止期、活动期系统性红斑狼疮病人（SLE）外周血单个核细胞（PBMC）环孢素A受体（CyP）mRNA表达及糖皮质激素对其表达的影响,为临床应用环孢素A辅助治疗该病提供理论依据。方法 采用逆转录PCR方法,经凝胶图像半定量分析,检测患者外周血单个核细胞CyP mRNA的表达。结果 SLE病人外周血单个核细胞存在有CyPmRNA的表达,静止期较正常人差异无显著性（P＞0．05）意义,但在活动期CyPmRNA的表达较正常人和静止期病人明显降低（P＜0．01）,地塞米松处理后,活动期病人CyPmRNA的表达水平显著上升（P＜0．01）。结论 SLE病人有CyPmRNA的表达,但活动期明显下降,糖皮质激素可改善CyP mRNA的表达。     

系统性红斑狼疮患者中性激素对淋巴细胞功能影响的研究

目的：通过检测SLE患者外周血淋巴细胞ERα的表达及外周血淋巴细胞功能的变化，探讨SLE发生过程中性激素对淋巴细胞功能的影响。方法：收集33例SLE患者和31例正常对照者血样，采用ELISA法检测外周血雌二醇（Estradiol，E2）、睾酮（Testosterone，T）水平，MTT法检测ConA刺激T淋巴细胞增殖反应，RT-PCR法检测外周血单个核细胞（Peripheral blood mononuclear cells，PBMCs）、T淋巴细胞和B淋巴细胞雌激素受体α（Estragen receptor，ERα）mRNA的表达。结果：与健康对照者相比较，SLE患者外周血T淋巴细胞增殖的刺激指数（Stimulatingindex，SI）降低（P〈0．01），E2水平升高，T降低，但P〉0．05，E2／T比值明显升高（P〈0．01），淋巴细胞ERαmRNA的表达均明显升高（P〈0．01）。结论：SLE患者性激素水平的改变可通过淋巴细胞ERα，影响淋巴细胞的功能，参与SLE的发生。     

细胞因子对Graves病甲状腺细胞凋亡及相关蛋白表达的影响

目的：探讨细胞因子（TNF-α、IL-1）诱导Graves病甲状腺细胞凋亡及细胞凋亡相关蛋白表达并与Graves病（GD）发病机制的关系。方法：免疫组化检测50例Graves病甲状腺组织的Fas表达。甲状腺组织取自GD手术标本，采用原代细胞培养方法。采用ELISA法检测细胞培养液中sFas含量。采用RT-PCR方法检测Fas／sFas mRNA。细胞因子诱导Graves病甲状腺细胞凋亡为观察组，设正常人为对照组。结果：观察组和对照组比较细胞凋亡率，有显著性的差异（P〈0．01）。观察组甲状腺细胞sFas、Fas／sFas mRNA含量与对照组比较明显增高（P〈0．01）。结论：细胞因子诱导Graves病甲状腺细胞凋亡及凋亡相关蛋白sFas、Fas／sFas mRNA有一定水平的表达，这些改变可能是细胞因子破坏甲状腺细胞的重要机制之一。     

CpG寡脱氧核苷酸对外周血单个核细胞功能的影响：1型糖尿病患者与健康者对照

背景：有研究表明CpG寡脱氧核苷酸可增强外周血单个核细胞的功能,但对1型糖尿病患者的影响至今少有报道。 目的：观察CpG寡脱氧核苷酸对1型糖尿患者患者与健康志愿者外周血单个核细胞γ干扰素、白细胞介素12,10表达的影响。 方法：将1型糖尿病患者与健康志愿者的外周血单个核细胞根据刺激物不同分为空白对照组、CpG寡脱氧核苷酸组。用RT-PCR法检测外周血单个核细胞γ干扰素、白细胞介素12 mRNA和白细胞介素10 mRNA的表达。 结果与结论：1型糖尿患者γ干扰素和白细胞介素12 mRNA的表达明显低于健康志愿者(P < 0.01)。1型糖尿患者与健康志愿者外周血单个核细胞经CpG寡脱氧核苷酸组刺激后,γ干扰素和白细胞介素12 mRNA的表达增高(P < 0.01),白细胞介素10 mRNA的表达无差异(P > 0.05)。结果提示,CpG寡脱氧核苷酸可促进1型糖尿患者外周血单个核细胞表达γ干扰素和白细胞介素12。     

多发性肌炎/皮肌炎患者外周血单个核细胞中糖皮质激素受体α、βmRNA表达与糖皮质激素疗效相关性的研究

目的探讨多发性肌炎/皮肌炎（PM/DM）患者外周血单个核细胞（PBMC）中糖皮质激素受体（GR）正常表位α和突变体表位β mRNA表达与PM/DM患者对糖皮质激素（GC）疗效的相关性。方法采用逆转录-聚合酶链反应（RT-PCR）方法对18例GC治疗敏感、10例GC治疗抵抗的PM/DM患者PBMC中GRα、β mRNA的表达水平进行了检测,同时采用放射免疫分析法测定血清中的皮质醇含量,并与20例正常对照进行比较。结果血清皮质醇含量在GC治疗敏感和抵抗的PM/DM患者及正常对照间差异均无统计学意义（均P〉0.05）。GC治疗敏感和抵抗的PM/DM患者PBMC中GRα mRNA的表达水平均显著低于正常对照（均P〈0.01）,且GC治疗抵抗的PM/DM患者显著低于GC治疗敏感的PM/DM患者（P〈0.05）。GC治疗敏感和抵抗的PM/DM患者PBMC中GRβ mRNA的表达均显著高于正常对照（均P〈0.05）,但在GC治疗敏感和抵抗的PM/DM患者间则差异无统计学意义（P〉0.05）。结论GRα mRNA表达水平降低可能与PM/DM患者对GC治疗抵抗相关,GRβ mRNA表达水平在PM/DM患者的GC治疗抵抗中无显著作用。     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.     

Seasonal variations in acute toxoplasmosis in pregnant women in Slovenia

Between December 1999 and December 2004, 40 081 pregnant women were examined for toxoplasmosis with Toxo-IgG, Toxo-IgM enzyme immunoassay. Women with positive results were then retested with the Toxo-IgG avidity assay for recent toxoplasmosis. Recent acute toxoplasmosis in pregnant women was found to be significantly more frequent (p < 0.01) during winter than summer. The incidence of acute toxoplasmosis during winter-spring was also significantly more frequent (p < 0.025) than summer-autumn. This phenomenon should be taken into account when formulating preventive measures for toxoplasmosis, especially for pregnant women.     

Age-related changes in polyamines in memory-associated brain structures in rats

Polyamines putrescine, spermidine and spermine are positively charged aliphatic amines and have important roles in maintaining normal cellular function, regulating neurotransmitter receptors and modulating learning and memory. Recent evidence suggests a role of putrescine in hippocampal neurogenesis, that is significantly impaired during aging. The present study measured the polyamine levels in memory-related brain structures in 24- (aged), 12- (middle-aged) and 4- (young) month-old rats using liquid chromatography/mass spectrometry and high performance liquid chromatography. In the hippocampus, the putrescine levels were significantly decreased in the CA1 and dentate gyrus, and increased in the CA2/3 with age. Significant age-related increases in the spermidine levels were found in the CA1 and CA2/3. There was no difference between groups in spermine in any sub-regions examined. In the parahippocampal region, increased putrescine level with age was observed in the entorhinal cortex, and age did not alter the spermidine levels. The spermine level was significantly decreased in the perirhinal cortex and increased in the postrhinal cortex with age. In the prefrontal cortex, there was age-related decrease in putrescine, and the spermidine and spermine levels were significantly increased with age. This study, for the first time, demonstrates age-related region-specific changes in polyamines in memory-associated structures, suggesting that polyamine system dysfunction may potentially contribute to aged-related impairments in hippocampal neurogenesis and learning and memory.     

休克过程中肾上腺髓质素变化的研究进展

Adrenomedullin (AM) is a new peptidergic regulator of vascular function. AM serves as a hormone, which has many biological properties, plays an important role in the many pathophysiological processes, especially shock. This review will highlight the structure, biological properties of AM and the relationship between AM and shock.     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Pitfalls in TRAP assay in routine detection of malignancy in effusions

Telomerase has been found to be reactivated in a majority of cancers but is inactive in most somatic cells. Our principal goal was to determine the potential use of the telomeric repeat amplification protocol (TRAP) assay as marker for malignancy in cytological effusions. The simple selection criterion was the cytological diagnosis, and routine samples were classified into malignant (58 samples) and nonmalignant (233 samples). Of the malignant samples, 44/58 (76%) were positive by TRAP assay. Of the 14 telomerase-negative cytology-positive samples, RNA integrity was poor in 9, indicating suboptimal sample conservation for molecular analysis. In 3 of the remaining 5 samples with a negative TRAP assay, a high number of malignant cells was observed, and these cells might have been telomerase-negative. Thus, the sensitivity of TRAP assay for the presence of malignant cells was about 76%. In the cytologically nonmalignant effusions, the presence of telomerase activity was observed in 24% (55/233). Of these, 6% were highly suspicious for malignancy, 9% were doubtful, and 9% were cytologically nonmalignant effusions confirmed by a follow-up of 12 mo or more. According to these data, the specificity of the TRAP assay to detect tumor cells in effusions ranged only between 82-91%. Our results indicate that, although the TRAP assay is positive in 6-15% of putative malignant effusions, the relatively high number of TRAP false-negative and false-positive cases renders this test unsuitable for routine diagnostic purposes.