Extracellular matrix activity and caveolae events contribute to cell surface receptor activation that leads to MAP kinase activation in response to UV irradiation in cultured human keratinocytes

Activation of cell surface components has been implicated in the activation of downstream signaling cascade in response to UV irradiation, and yet the identity and the interaction of those components have been scantly documented. Accumulating evidence indicates that caveolae encapsulating caveolins is the location for those interactions. We found in cultured human keratinocytes that UV irradiation induced both caveolin-1 and EGFR phosphorylation. Filipin, a caveolae disruptive agent, inhibited UV-induced caveolin-1 activation. Na+-K+-ATPase catalyzes active transport of Na+ and K+ across plasma membrane of mammalian cells, inactivation of which has recently been shown to be involved in the activation of signal transduction pathways including MAP kinase cascade. We found in this study that UV inactivated Na+-K+-ATPase in time-dependent manner, Na+-K+-ATPase activity started to decrease 5 min post UV irradiation and reduced to 60% of its original activity within 1 h. Pretreatment with Flipin and MMP inhibitor recovered Na+-K+-ATPase activity lost by UV irradiation. ECIS analysis indicated that both EGF treatment and UV irradiation increased membrane electric activity which was inhibited by MMP inhibitor and Filipin. Further study showed that pretreatment of human keratinocytes with MMP inhibitor or Filipin inhibited UV-induced phosphorylation of p38 and JNK, which was however not observed in LnCap cells, a prostate cancer cell line lacking caveolin-1. UV irradiation also induced ectodomain shedding of HB-EGF in a time-dependent manner in keratinocytes. Collectively, we conclude that UV-induced MAP kinase activation is mediated by cell surface receptor activation due to the matrix activity and membrane caveolae function and inactivation of Na+-K+-ATPase.     

遗传算法在256阵元相控阵高强度聚焦超声治疗系统多焦点控制中的应用

阐述了遗传算法优化多点超声聚焦的方法以及球面相控阵声场计算方法。对实验室研发的256阵元相控阵聚焦超声治疗系统作了简介。利用此遗传算法和声场计算方法，对轴上和离轴单焦点以及轴对称六焦点和非轴对称四焦点进行了256阵元相控阵的声场仿真，并观察了实验室研发的256阵元相控阵聚焦超声治疗系统作用于有机玻璃和透明仿体的实验结果。用遗传算法和声场计算方法的仿真和系统实验结果表明，此方法可在实际聚焦手术中准确地控制3维单焦点和3维多焦点。     

Microsatellite alterations in hepatocellular carcinoma and intrahepatic cholangiocarcinoma

A series of 20 hepatocellular carcinomas and 8 intrahepatic cholangiocarcinomas was screened from the Korean population for microsatellite alterations, including a loss of heterozygosity and replication errors using nine microsatellite markers containing several genes. The microsatellite results and our previous comparative genomic hybridization results of two tumors were compared at each locus, and the correlations between these and clinicopathologic variables were examined. The most characteristic findings were found at 13q. Replication errors were prevalent at D13S160 (13q21.2 approximately q31) and D13S292(13q12). The incidence of loss of heterozygosity, however, was higher at D13S153 (13q14.1 approximately q14.3) and D13S265(13q31 approximately q32). In contrast, there were higher deletion frequencies observed in hepatocellular carcinoma (HCC) and higher amplification frequencies observed in intrahepatic cholangiocarcinoma at 13q in our previous comparative genomic hybridization (CGH) study. Higher frequencies of replication errors were observed at D16S408 (13q12 approximately q21) and D16S504(13q23 approximately q24) in the HCC. This study found that significant differences in the patterns of genetic instability of microsatellites were dependent on the chromosomal loci. It is believed that certain genes at altered CGH regions, which are relevant to the development and/or progression of these cancers, are activated by different mutation mechanisms.     

Gemins modulate the expression and activity of the SMN complex

Reduction in the expression of the survival of motor neurons (SMN) protein results in spinal muscular atrophy (SMA), a common motor neuron degenerative disease. SMN is part of a large macromolecular complex (the SMN complex) that includes at least six additional proteins called Gemins (Gemin2-7). The SMN complex is expressed in all cells and is present throughout the cytoplasm and in the nucleus where it is concentrated in Gems. The SMN complex plays an essential role in the production of spliceosomal small nuclear ribonucleoproteins (snRNPs) and likely other RNPs. To study the roles of the individual proteins, we systematically reduced the expression of SMN and each of the Gemins (2-6) by RNA interference. We show that the reduction of SMN leads to a decrease in snRNP assembly, the disappearance of Gems, and to a drastic reduction in the amounts of several Gemins. Moreover, reduction of Gemin2 or Gemin6 strongly decreases the activity of the SMN complex. These findings demonstrate that other components of the SMN complex, in addition to SMN, are critical for the activity of the complex and suggest that Gemin2 and Gemin6 are potentially important modifiers of SMA as well as potential disease genes for non-SMN motor neuron diseases.     

从人外周血B淋巴细胞中应用PCR扩增人抗体基因

本文用常规ＰＣＲ法和半套式ＰＣＲ法，以一组人抗体重链和轻链引物，直接从人外周血淋巴细胞中扩增出抗体重链Ｆｄ基因和轻链基因。一些常规ＰＣＲ法不能直接扩增的人抗体基因，用半套式ＰＣＲ法扩增却得到了阳性结果。扩增的抗体基因的分子量与国内外同类报道一致。本文结果提示，在建立抗体基因文库时，半套式ＰＣＲ法能进一步丰富扩增的抗体基因的多样性     

从EST克隆中筛选肝癌内高表达的基因片段

应用Ｎｏｒｔｈｅｒｎ杂交技术，从表达序列标记（Ｅｘｐｒｅｓｓｅｄｓｅｑｕｅｎｃｅｔａｇ，ＥＳＴ）克隆中筛选出一个在肝癌组织内高表达，而在相应癌旁肝及正常肝组织内低表达或不表达的基因片段。ＤＮＡ序列测定表明，该基因片段为一未知新基因的部分序列。此基因片段可作为探针，进一步筛选ｃＤＮＡ文库，以得到新的癌基因的候选基因。     

Recalling sexual partners: the accuracy of self-reports

Accuracy of recall of the number of sexual partners individuals had over a period of one month, three months, six months and one year was studied in a group of 285 young, single, heterosexual adults. Self-reports of the number of partners were obtained on a weekly basis and then compared with recall of behavior over longer time periods that overlapped the weekly measures. For individuals who claimed abstinence or who claimed to be monogamous, accuracy of recall was relatively high, especially at the shorter time frames. Level of education was related to accuracy for claimed abstainers, such that lower levels of education were associated with lower accuracy of recall. Accuracy rates for individuals who reported having multiple sexual partners tended to be lower and were found to be related to one's propensity to engage in casual sex.     

肾细胞癌中c-erbB-2表达及其意义

目的子研究c—erbB-2在肾细胞癌中的表达及其与临床分期、病理分型、病理分级和预后的关系。方法 应用免疫组化S-P法分别用鼠抗人c-erbB-2胞内段单克隆抗体(CB11)和鼠抗人c-erbB-2胞外段单克隆抗体(9G6．10)检测77例肾细胞癌及相应癌旁肾组织c—erbB-2蛋白表达。应用RT-PCR方法检测10例新鲜肾癌组织及癌旁正常组织c—erbB-2 mRNA的表达。透明细胞癌、颗粒细胞癌及乳头状肾细胞癌c—erbB-2的表达分别为68．2％、93．1％及75．0％。结果 77例肾癌组织CB11和9G6．10的阳性率分别为61%(47／77)和45．5%(35／77)；二者联合检测总阳性率为77．9％(60／77)。透明细胞癌、颗粒细胞癌及乳头状肾细胞癌c—erbB-2的表达分别为68．9％、92．9％及75%。RT-PCR检测新鲜肾癌组织c—erbB-2mRNA的阳性率为100％(10／10)。结论 肾癌中c—erbB-2蛋白及c—erbB-2mRNA均过表达。不同的肾癌病理类型c—erbB-2蛋白表达不同，颗粒细胞癌表达最高。c—erbB-2在肾癌的不同临床分期均过表达，说明可能在病变的早期c—erbB-2已经发生改变。c-erbB-2的表达与临床分期及肿瘤分级无明显相关性。     

SARS病毒S1蛋白重组C端片段免疫效果的实验研究

为获得纯化的重组SARS病毒S1蛋白C端 ,研究其刺激机体产生针对SARS病毒免疫应答的规律和机制 ,将编码SARS病毒S1蛋白C端 311个氨基酸残基的基因克隆 ,并在原核表达系统中表达 ,纯化获得了的重组蛋白。利用SARS患者恢复期的血清 ,对纯化的重组S1蛋白进行血清学分析。结果表明 ,本研究中克隆表达的重组蛋白序列与公布的SARS病毒S1蛋白C端的序列相同 ,其编码的重组蛋白相对分子质量约为 5 90 0 0Mr。SARS患者恢复期的血清均与重组蛋白反应 ,在5 9 0 0 0Mr处形成特异性的反应条带 ,而来自SARS流行前的正常人对照血清则不能与重组蛋白反应。在本研究中获得的重组蛋白可以为研究SARS病毒识别宿主细胞受体的过程及其机制提供条件。