Metabolic activation of methyl-hydroxylated derivatives of 7,12- dimethylbenz[a]anthracene by human liver dehydroepiandrosterone-steroid sulfotransferase

Methyl-hydroxylated metabolites of the potent carcinogen, 7,12- dimethylbenz[a]anthracene (DMBA), namely, 7-hydroxymethyl-12- methylbenz[a]anthracene (7-OH-DMBA), 7-methyl-12- hydroxymethylbenz[a]anthracene (12-OH-DMBA) and 7,12- dihydroxymethylbenz[a]anthracene (7,12-diOH-DMBA), were examined as substrates for sulfotransferase bioactivation in different human tissue cytosols. Hepatic cytosols, which were able to catalyze the 3'- phosphoadenosine 5'-phosphosulfate (PAPS)-dependent DNA binding of 7-OH- DMBA, 12-OH-DMBA and 7,12-diOH-DMBA, were highly sensitive to inhibition by dehydroepiandrosterone (DHEA), a specific substrate for human DHEA-steroid sulfotransferase (IC50 = 5 microM). By comparison, 2,6-dichloro-4-nitrophenol, a potent inhibitor of the thermostable (TS)- phenol and estrogen sulfotransferases, did not have an appreciable inhibitory effect. Neither p-nitrophenol, a high affinity substrate for human TS-phenol and estrogen sulfotransferases, nor dopamine, a specific substrate for the thermolabile (TL)-phenol sulfotransferase, significantly inhibited the DNA binding of 12-OH-DMBA catalyzed by hepatic cytosols. Inter-subject variation (n = 12) of the PAPS- dependent DNA binding of 12-OH- and 7,12-diOH-DMBAs also correlated well with DHEA-sulfotransferase activity (r = 0.90; P < 0.00001 and r = 0.92; P < 0.00001, respectively). This sulfation-dependent metabolic activation was not detected in cytosols from human colon, pancreas, larynx or mammary gland. Both TS- and TL-phenol sulfotransferases were active in human liver and colon but only liver contained DHEA- sulfotransferase activity. These results indicate that the sulfotransferase-mediated activation of the methyl-hydroxylated DMBAs is predominantly catalyzed by DHEA-steroid sulfotransferase in human liver and that TS- and TL-phenol sulfotransferases and estrogen sulfotransferase are not involved in the catalysis.      

Monoclonal antibodies to mammalian heat shock proteins impair mouse embryo development in vitro

Two-cell mouse embryos (B6D2F1) were cultured in the presence or absence of 100 microg/ml monoclonal antibodies specific for the mammalian 60 kDa (HSP60), 70 kDa (HSP70) and 90 kDa (HSP90) heat shock proteins. Embryo development was evaluated after 3, 5 and 7 days in culture by determining the number of blastocysts, hatched blastocysts and outgrown trophoblasts at the successive time points. At day 3, only 29% (22/75) of the embryos cultured with anti-HSP60 antibody developed to the blastocyst stage (P < 0.0001) as compared to 67% (31/46) of the embryos cultured with anti-HSP70, 72% (43/60) cultured with anti-HSP90, and 79% (49/62) in medium plus mouse IgG1. By day 5, hatched embryos were present in 28% (13/ 46) of the cultures containing anti-HSP70 (P < 0.0001), as opposed to 57% (34/60) containing anti-HSP90 and 73% (45/62) containing IgG1. At day 7, outgrown trophoblasts were observed in 9% (4/46) of cultures containing anti-HSP70 (P < 0.0001), 45% (27/60) containing anti-HSP90 (P < 0.01) and 66% (41/62) cultured in medium plus IgG1. Antibodies to different heat shock proteins exerted a detrimental effect on mouse embryo development at unique development stages. Immune sensitization to heat shock proteins may be a cause of reproductive failure.      

First trimester development of human chorionic villous vascularization studied with CD34 immunohistochemistry

Normal chorionic villous vascularization is essential for the undisturbed development of pregnancy. Defective vasculogenesis may play a role in pathological pregnancy. To assess pathological chorionic villous vascularization, normal vascularization has to be defined first. Few data are available on this topic. The aim of this study was therefore to investigate normal chorionic villous vascularization in ultrasound-dated first trimester pregnancies from week 5 menstrual age to week 12 (n = 41), using quantitative CD34 immunohistochemistry. Two important processes in chorionic villous vascularization were quantitatively illustrated: (i) maturation, reflected by an increase of the total number of luminized vessels as opposed to non-luminized haemangioblastic cords and (ii) margination, due to a decrease of villous stromal area and an increase of total villous vascular area. The percentage of villous stromal area occupied by vascular elements (area difference %) increased from 0.7% in week 5-2.5% in week 10. Therefore, the area of the villous stroma occupied by vascular elements increases and the vessels are situated closer to the trophoblastic layer suitable for fetal-maternal exchange. There was also a trend in increased number of peripheral vessels (2.0 in week 5 to 4.6 in week 10), supporting both developmental mechanisms. In conclusion, in exactly dated normal human first trimester pregnancies, development of the chorionic villous vascular system seems to be mostly characterized by maturation of luminized vessels from primitive haemangioblastic cords, and margination to a situation of peripherally located vessels.      

血清反复冻融对HBsAg和抗HBs抗体检测的影响

0　引言　 EL ISA技术已在乙肝标志物的检测中已普遍应用 ,一般认为收集的血清应及时检测 .而在基层单位或在流行病学调查研究中 ,由于血标本收集不集中 ,或者有些标本常需检测多个指标而持续时间较长 ,因此 ,血清必须低温贮存一定时间或反复冻融后检测 ,而血清冻融对 EL ISA检测结果是否有影响尚不清楚 .为此 ,我们对 183份血清标本冻融前、冻融 3次、冻融 6次后的 HBs Ag和抗 - HBs进行了检测和分析比较 .1　材料和方法1.1　材料　 1999- 0 5收集西京医院门诊乙肝 5项检测血清183份 ,年龄、性别、诊断不限 .1.2　方法　于初次检测 …     

Close mapping of the focal non-epidermolytic palmoplantar keratoderma (PPK) locus associated with oesophageal cancer (TOC)

Focal non-epidermolytic palmoplantar keratoderma (PPK or palmoplantar ectodermal dysplasia type III) is associated with oesophageal cancer in three families: two large pedigrees located in Liverpool, UK and in the midwestern American states and one smaller family from Germany. In these families, the PPK is inherited as autosomal dominant and has a late onset, usually manifesting between 7 and 8 years of age. The disease is characterised by thickening of the pressure areas of the soles, but is not restricted to the feet and also presents with oral leukokeratosis and follicular hyperkeratosis. The disease locus [previously termed the "tylosis oesophageal cancer gene' (TOC) locus] has been mapped to 17q23-qter by linkage analysis. This region is located telomeric to the keratin 16 gene, in which mutations have been identified in focal PPK families who show no increased cancer risk. We describe the close mapping of this locus to the interval between AFMb054zf9 and D17S1603 using haplotype analysis of additional Genethon markers in the region and show that although the American family is unlikely to be related to either of the other two, the UK and German pedigrees may share a common descent. This work provides a basis for positional cloning and candidate gene analysis in order to identify a gene that may be involved in familial oesophageal cancer.      

成纤维细胞的力学生物学(上)

由于重力、血液的流动及运动的原因，人体会不断受到力的作用。众所周知，结缔组织是承受与传导力的器官，其内的细胞可以通过多种机制将力转化为生物化学信号，但是有些机制尚未完全了解。结缔组织的成纤维细胞就是对力产生反应的细胞，在力的作用下，会通过改变自身细胞外基质（extracellular matrix，ECM）的基因及蛋白质表达的方法维持组织的结构和功能（如创伤修复）。当结缔组织承受到较大的压力时，可以使结缔组织保持正常的功能和组织的动态平衡。结缔组织的修复及维持主要由间充质细胞或成纤维细胞来完成。力可以调节细胞的多种功能，如细胞增殖、基因表达及蛋白质分泌。