大鼠肺微血管内皮细胞培养及其粘弹性研究

为了建立肺微血管内皮细胞培养方法 ,研究肺微血管内皮细胞粘弹性。我们取大鼠肺周边组织 (宽度不应大于 1.5 mm) ,将组织剪成 1.5 mm× 1mm× 1mm的组织块 ,贴入无菌的 2 5 cm3培养瓶 ,每瓶 10～ 15块 ,同时加入含 2 0胎牛血清、肝素 90 U/ml、L-谷氨酰胺 4mmol、青霉素 10 0 U/ml和链霉素 10 0 μg/ml的 DMEM培养基 3ml,放入 37℃二氧化碳培养箱中静置培养 ;8h后翻转培养瓶 ,6 0 h后取出肺组织块 ,接着继续培养 2～ 4d后进行传代。最后消化分离肺微血管内皮细胞 ,用微管吸吮系统研究肺微血管内皮细胞粘弹性。结果显示 :肺微血管内皮细胞通过倒置相差显微镜观察 ,细胞呈鹅卵石镶嵌状排列 ,状如梭形或多角形 ,大小均匀 ,胞核清晰 ,呈卵圆形 ,胞浆丰富 ; 因子相关抗原免疫荧光染色呈阳性 ;肺微血管内皮细胞弹性模量 K1 =49.3± 9.2 Pa、K2 =73.2±2 4.8Pa、粘性系数 μ=19.2± 7.2 Pa.s。这些结果表明用组织块法培养肺微血管内皮细胞是可行的 ,肺微血管内皮细胞表现出较大的刚性     

Polymorphism of human leukocyte antigen-DRB1, -DQB1, and -DPB1 genes of Shandong Han population in China

In the present study, polymerase chain reaction-sequence-based typing (PCR-SBT) was used to analyze human leukocyte antigen (HLA)-DRB1, -DQB1, and -DPB1 alleles of 98 unrelated healthy Shandong Han individuals. A total of 60 alleles, in which 28 in DRB1, 15 in DQB1 and 17 in DPB1 were found. Among the 28 detected DRB1 alleles, DRB1*150101, DRB1*070101, DRB1*090102, DRB1*120201, and DRB1*080302 were commonly observed, with frequencies of 16.3%, 11.2%, 10.2%, 8.2%, and 5.6%, respectively. The most predominant DQB1 allele was DQB1*030101/0309 with the frequency of 20.4%, followed by DQB1*0201/0202 (14.8%), DQB1*0602 (14.3%), DQB1*030302 (12.2%), and DQB1*060101/060103 (10.7%). Of the 17 detected DPB1 alleles, DPB1*0501 was the most frequent allele with the frequency of 37.2%. DPB1*020102 (18.4%), DPB1*040101 (11.2%), DPB1*0402 (7.1%), and DPB1*1701 (6.6%) were also very frequent alleles. A total of 53 estimated DRB1-DQB1 two-locus haplotypes were observed in Shandong Han population, of which DRB1*150101-DQB1*0602 was the most predominant, followed by DRB1*090102-DQB1*030302, DRB1*070101-DQB1*0201/0202 DRB1*120201-DQB1*030101/0309, and DRB1*080302- DQB1*060101/060103. The distribution of the HLA class II alleles and haplotypes frequencies as well as the dendrogram showed that the Shandong Han population belongs to the northern group of Chinese. The data have implications for anthropological studies and disease associations.     

恶性上皮性肿瘤中CK20的表达及其临床意义

目的　研究单克隆抗体CK2 0在恶性上皮性肿瘤和卵巢转移性腺癌组织中的表达及其意义。方法　应用S P法对鼻咽非角化性癌、乳腺浸润性导管癌、肺的鳞癌和腺癌、卵巢黏液性囊腺癌、胃腺癌和结肠直肠腺癌各组总计 6 7例和 4 1例分别进行了CK2 0和CK19检测。结果　CK2 0阳性率 :肺腺癌 1/ 7(14 3% ) ,卵巢浆液性和黏液性腺癌 3/ 12 (33 3% ) ,胃腺癌 3/ 9(33 3% ) ,结肠直肠腺癌组 2 1/ 2 2 (95 5 % ) ,其他癌组织均呈阴性。结肠直肠腺癌组组与其他各组间比较差异有显著性 (P <0 0 1)。CK19在上述 4 1例癌组织中均呈强阳性表达。结论　CK2 0表达对鉴别结肠腺癌和直肠腺癌与肺腺癌和乳腺浸润性导管癌具有高度特异性和较高的敏感性 ;CK2 0高表达对鉴别卵巢原发性腺癌与卵巢的结肠腺癌或直肠腺癌转移具有一定的意义     

Characterization of a novel HLA-DQB1 allele, DQB1*020102

A novel human leukocyte antigen (HLA)-DQB1 allele, DQB1*020102, was detected in a 28-year-old woman of Han ethnic in Guangzhou, China. Compared with HLA-DQB1*020101 and HLA-DQB1*0202, they differed in only one nucleotide at the position 167 (C to T) of exon 2, which was a highly conserved position. This is a synonymous mutation, which does not cause any change in the amino acid sequence of mature protein.     

高校学生朋友心理辅导员培训内容与方法研究

目的 学生朋友心理辅导员是高校学生心理健康教育网络系统的重要组成部分。如何在较短的时间里，经过比较系统的培训，让普通的大学生成长为合格的学生朋友心理辅导员并承担起为同学进行心理护理的义务，是构建高校学生心理健康教育网络系统的关健。方法 明确培训目标和担任工作职能，培训内容科学，知识构成合理，培训方法灵活，进行培训前测与后测和培训效果评估，注重实践锻炼和心理督导。结果 培训结业的学员在各方面有比较真显著变化，心态明显改变，知识明显提高、助人技巧明显增强、人际交往明显改善，总体上呈现出积极、友爱、乐观、向上的精神面貌。结论 在高校能够培训出合格的学生朋友心理辅导员，并成为高校学生心理健康教育网络的重要力量。     

细胞性一氧化氮供体的建立及其对肿瘤细胞凋亡的诱导效应

目的一氧化氮（ＮＯ）供体细胞系的建立及其对肿瘤细胞凋亡的诱导效应研究。方法采用３Ｈ－胍氨酸转换法进行ＮＯ合成酶（ＮＯＳ）活性测定；亚硝酸盐检测；ＤＮＡ片段的提取及凝胶电泳分析；凋亡细胞的ＤＡＰＩ染色观察；Ｗｅｓｔｅｒｎｂｌｏｔ分析。结果将ｉＮＯＳ真核表达质粒、ｐＣＭＶ／ｉＮＯＳ转染至Ｓｐ２／０骨髓瘤的变异株中，并获得能稳定表达ｉＮＯＳ和合成ＮＯ的重组细胞系（ＳＰｍｔ／ｉＮＯＳ），表达ｉＮＯＳ的效率为每升培养物含４００μｇ蛋白。利用该细胞作ＮＯ细胞性供体，成功地证实ＮＯ能诱导Ｓｐ２／０骨髓瘤细胞发生凋亡。结论所建细胞性ＮＯ供体应用于ＮＯ的生物学功能研究具有ＮＯ合成稳定；更能模拟体内细胞间ＮＯ的信息传递过程；不需任何外源刺激即可合成ＮＯ等独特优点。所建ＮＯ供体细胞可以用于肿瘤细胞凋亡的研究。     

Morphometric analysis of the immunohistochemical expression of Clara cell 10-kDa protein and surfactant apoproteins A and B in the developing bronchi and bronchioles of human fetuses and neonates

 Morphometric analyses of the immunohistochemical expression of the Clara cell secretory 10-kDa protein (CC10) and surfactant apoproteins A and B (SP-A and -B) were carried out on the developing bronchi and bronchioles of human fetuses and neonates. We analysed the ratio of the number of CC10-positive cells per subepithelial length of the bronchial or bronchiolar basement membrane and found that both the bronchial and the bronchiolar population of CC10-positive cells was significantly higher than that of either SP-A or SP-B. In addition, CC10 was found to be distributed mainly in the bronchiole. CC10-positive cells began to be recognized in the late pseudoglandular phase (15 weeks of gestation) and thereafter gradually increased in the canalicular and terminal sac phases, which correspond to the active development period of the acini or peripheral airways. The earliest expression of SP-A was also noted at 15 weeks of gestation, but its positive epithelial cells were present mainly in the larger bronchi. Double immunohistochemical staining for CC10 and SP-A revealed that the CC10-positive cells lining both the bronchi and bronchioles were different from the SP-A-positive cells. This finding suggests that CC10-positive cells are functionally and developmentally heterogeneous in both fetal and neonatal lungs in humans Received: 22 May 1997 / Accepted: 21 July 1997     

Pds1 phosphorylation in response to DNA damage is essential for its DNA damage checkpoint function

In Saccharomyces cerevisiae, Pds1 is an anaphase inhibitor and plays an essential role in DNA damage and spindle checkpoint pathways. Pds1 is phosphorylated in response to DNA damage but not spindle disruption, indicating distinct mechanisms delaying anaphase entry. Phosphorylation of Pds1 is Mec1 and Chk1 dependent in vivo. Here, we show that Pds1 is phosphorylated at multiple sites in vivo in response to DNA damage by Chk1. Mutation of the Chk1 phosphorylation sites on Pds1 abolished most of its DNA damage-inducible phosphorylation and its checkpoint function, whereas its anaphase inhibitor functions and spindle checkpoint functions remain intact. Loss of Pds1 phosphorylation correlates with APC-dependent Pds1 destruction in response to DNA damage. We also show that APC(Cdc20) is active in preanaphase arrested cells after DNA damage. This suggests that Pds1 is stabilized by phosphorylation in response to DNA damage, but APC(Cdc20) activity is not altered. Our results indicate that phosphorylation of Pds1 by Chk1 is the key function of Chk1 required to prevent anaphase entry.     

Role of glutathione metabolism of Treponema denticola in bacterial growth and virulence expression

Hydrogen sulfide (H(2)S) is a major metabolic end product detected in deep periodontal pockets that is produced by resident periodontopathic microbiota associated with the progression of periodontitis. Treponema denticola, a member of the subgingival biofilm at disease sites, produces cystalysin, an enzyme that catabolizes cysteine, releasing H(2)S. The metabolic pathway leading to H(2)S formation in periodontal pockets has not been determined. We used a variety of thiol compounds as substrates for T. denticola to produce H(2)S. Our results indicate that glutathione, a readily available thiol source in periodontal pockets, is a suitable substrate for H(2)S production by this microorganism. In addition to H(2)S, glutamate, glycine, ammonia, and pyruvate were metabolic end products of metabolism of glutathione. Cysteinyl glycine (Cys-Gly) was also catabolized by the bacteria, yielding glycine, H(2)S, ammonia, and pyruvate. However, purified cystalysin could not catalyze glutathione and Cys-Gly degradation in vitro. Moreover, the enzymatic activity(ies) in T. denticola responsible for glutathione breakdown was inactivated by trypsin or proteinase K, by heating (56 degrees C) and freezing (-20 degrees C), by sonication, and by exposure to N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK). These treatments had no effect on degradation of cysteine by the purified enzyme. In this study we delineated an enzymatic pathway for glutathione metabolism in the oral spirochete T. denticola; our results suggest that glutathione metabolism plays a role in bacterial nutrition and potential virulence expression.