Characterization of antimicrobial resistance among Escherichia coli O111 isolates of animal and human origin

Fifty isolates of Escherichia coli serogroup O111 recovered from humans and various animal species over a 24-year period (1976-1999) were examined for typical virulence-associated factors and susceptibilities to antimicrobials of human and veterinary significance. Nine H (flagellar) types were identified including nonmotile (n = 24), 32 (n = 12), negative (n = 5), and 56 (n = 3). Thirty-five (70%) isolates possessed at least one Shiga-toxin-producing E. coli (STEC)-associated virulence determinants (eae, stxl, stx2, hlyA) via PCR analysis. Of these 35 isolates, 20 possessed eae, stxl, and hlyA genes, whereas three isolates possessed eae, stxl, stx2, and hylA genes. Multiple antibiotic resistance was observed in 70% of the 50 E. coli O111 isolates. The majority of isolates displayed resistance to streptomycin, sulfamethoxazole, tetracycline, and kanamycin. Bacterial resistance to ampicillin, gentamicin, chloramphenicol, trimethoprim and apramycin was also observed. Integrons were identified in 23 (46%) of the E. coli isolates assayed, with a 1-kb amplicon being most frequently observed. DNA sequencing of these integrons revealed the presence of the aadA gene, encoding resistance to streptomycin. Two integrons of 1.5 and 2 kb contained the aadA2 and either dfrI or dfrXII genes, encoding resistance to streptomycin and trimethoprim, respectively. Integrons were also identified from isolates dating back to 1982. Isolates were further genetically characterized via ribotyping, which identified 15 distinct ribogroups, with 62% of isolates clustering into four major ribogroups. Certain riboprint patterns from different animal species, including humans, were observed in isolates spanning the 24-year collection period, suggesting the dissemination of specialized pathogenic O111 clones.     

Comparison of ethanol versus formalin fixation on preservation of histology and RNA in laser capture microdissected brain tissues

Although RNA can be retrieved from formalin-fixed, paraffin-embedded (FFPE) tissues, the yield is low, and the RNA is fragmented. Recent advances in gene expression profiling underscore the importance of identifying a fixative that preserves histology and mRNA. We demonstrated that, for immersion fixation of brains, 70% ethanol is superior to formalin for mRNA preservation. RNA yield from ethanol-fixed tissues was 70% of the yield from fresh frozen specimens, but only a negligible quantity was recovered from formalin-fixed tissues. RNA from ethanol-fixed brains showed integrity comparable to RNA from fresh frozen tissues, and RT-PCR using RNA from ethanol-fixed tissues was consistently successful. RNA from FFPE tissues composed of low-molecular weight fragments, and their use in RT-PCR failed repeatedly. The yield and quality of RNA from ethanol-fixed brains were unaffected after immersion at 4 degrees C for 2 weeks. In a blinded comparison to FFPE tissues, ethanol-fixed specimens were judged to show comparable histology and superior immunostaining. After laser capture microdissection (LCM), we failed to recover mRNA from FFPE tissues but retrieved mRNA from ethanol-fixed tissues for RT-PCR and cDNA microarray analysis. We conclude that 70% ethanol preserves RNA integrity and is suitable for expression profiling of brain tissues by LCM and cDNA microarray.     

GDNF-induced seminomatous tumours in mouse--an experimental model for human seminomas?

Glial-cell-line-derived neurotrophic factor (GDNF) is a distant member of the transforming growth factor superfamily. It binds to and activates a receptor complex consisting of GFR-alpha1 and Ret receptor tyrosine kinase. In testis, GDNF is expressed by Sertoli cells. We have shown by transgenic loss- and gain-of-function mouse models that GDNF regulates the cell fate decision of undifferentiated spermatogonia. In the GDNF +/- mice, the spermatogonia differentiate in excess leading to the depletion of germ cells. In the mice overexpressing GDNF in testes, undifferentiated spermatogonia accumulate in the tubules, no sperm is produced, and the mice are infertile. After a year, the GDNF overexpressing mice frequently (89%) develop testicular tumours, and most of them are bilateral (56%). All these tumours show the same histological pattern. They are composed of round spermatogonial/gonocytic cells with only a scant cytoplasm. The tumours are locally invasive but do not metastasise. They express germ line markers, are positive for alkaline phosphatase, and aneuploid with a triploid peak. Thus, by several histological, molecular, and histochemical characteristics, the GDNF-induced tumours mimic classical seminomas in men, but the precursor lesions are apparently different in mouse and man.     

用非同位素原位杂交组化在光镜和电镜水平观察前阿黑皮素mRNA在垂体的分布

用地高辛标记反意前阿黑皮素(pro-opiomelanocortin,POMC)cRNA探针原位杂交组化在光镜和电镜水平观察了POMC mRNA在大鼠垂体的分布,并比较了碱性磷酸酶(AKP)显色系统和辣根过氧化酶(HRP)显色系统在光镜水平上的敏感性.结果:POMC mRNA广泛地分布于垂体的中间叶和前叶.中间叶全部细胞均为POMC mRNA阳性.前叶中除前叶的腹侧缘和前叶与中间叶交界处阳性细胞较少外,都有较多的POMC mRNA阳性细胞分布.AKP显色系统比HRP显色系统敏感.在电镜水平,POMC mRNA主要分布于粗面内质网,少数分泌颗粒可能呈阳性反应,胞核未见阳性反应沉淀.文内还就阳性分泌颗粒在调节细胞合成功能方面可能起的作用进行了讨论.     

Adenovirus-mediated p53 gene therapy in nasopharyngeal cancer

EBV-associated nasopharyngeal cancer (NPC) occurs with high frequency in China and is a major cause of morbidity and mortality. To explore the potential use of adenovirus-mediated tumor suppressor p53 gene therapy In NPC, we first examined the in vitro effects of p53 introduced into the NPC cell lines RPMI 2650, Fadu and Detroit 562. p21(WAF1/CIP1) induction by chemotherapy was used as a functional assay which revealed that RPMI 2650 expresses wild-type p53 whereas Fadu and Detroit 562 encode mutant p53. Infection with p53-expressing adenovirus (Ad-p53) induced apoptosis and inhibited cell growth in all three NPC cell lines, regardless of the endogenous p53 status. Adenovirus infectivity was greatest in RPMI 2650 cells, with 100% of the cells expressing beta-galactosidase following Ad-LacZ infection using an MOI of 100, as compared to 20-30% infectivity with the other NPC lines. Using RPMI 2650 cells injected into nude mice, we developed an animal model for nasopharyngeal cancer. Established tumors (0.6-0.8 cm) were injected with 5x10(9) PFU Ad-LacZ, Ad-p53 or PBS in a 100 mu l volume. We found evidence for in vivo expression of beta-galactosidase or p53 and p21 up to two weeks following Ad-LacZ or Ad-p53 virus injection respectively. Objective regression of tumor size was observed at two weeks in 4/6 Ad-p53-treated tumors, but not in Ad-LacZ or PBS-treated tumors. The results provide an animal model for human nasopharyngeal cancer, and indicate a potential use of p53 in its therapy in vivo.     

Predicting the viability of grafted livers in rats through a rapid and sensitive metabolic indicator assessed by 31P-NMR spectroscopy

The present study was undertaken to clarify whether a correlation exists between the hepatic ratio of the -phosphorous moiety of ATP (-ATP) to inorganic phosphate (Pi), measured by ³¹P nuclear magnetic resonance spectroscopy 1 h after the reestablishment of portal blood flow, and the survival rate of rats following liver transplantation. This ratio was compared with the arterial ketone body ratio [AKBR (acetoacetate/3-hydroxybutyrate)], which is accepted as a reliable indicator of liver viability. After the transplantation of fresh livers, the 1-week survival rate was 92% and the -ATP/Pi ratio was 64% of the normal level. When the liver grafts were subjected to warm ischemia for 25 min or 45 min prior to harvesting, the 1-week survival rate decreased to 43% and 0%, respectively, and the -ATP/Pi ratio dropped to 31% and 18% of the normal level, respectively. On the other hand, the AKBR was about 25% of the normal level after transplantation of fresh livers, while it was 37% and 48% after transplantation with 25 min and 45 min of warm ischemia, respectively. However, 4h after the reestablishment of portal blood flow, the AKBR correlated with the -ATP/Pi ratio in both the fresh graft group and the 45-min warm ischemic damage group. These results show that the -ATP/Pi ratio provides an accurate evaluation of a graft viability even at an extremely early stage following liver transplantation, and should prove useful for the early diagnosis of primary graft nonfunction after liver transplantation.     

侧脑室—腹腔分流治疗脑积水62例报告

目的　探讨脑积水的治疗方法和疗效。方法　62例均行侧脑室-腹腔分流术。脑室端从枕角穿入10cm接引流泵,再从皮下隧道引至上腹或左下腹放入腹腔。结果　1～4周后复查,症状完全消失56例,部分改善5例,无变化1例。57例复查CT示:52例脑室恢复正常,4例脑室较前缩小,1例无变化。堵管9例。低颅内压2例,硬膜下血肿2例,经保守治疗痊愈。49例随访1～8年,41例能正常工作学习,5例生活自理,1例植物生存,2例死亡。结论　明确诊断,掌握手术适应症及手术技巧是成功的关键。     

3种低温保存板层角膜移植材料的电镜观察

评价低温保存板层角膜移植材料的效果。方法将１５只人尸角膜分三组保存２年：４℃甘油脱水法,角膜液氮冷冻法,全眼球液氮冷冻法。并做透射电镜观察。结果第一组中,角膜胶原纤维排列紊乱,疏松,缺乏连续性,可见广纤维变性,纤维间可见极低电子密度灶。     

应用Opalescence脱色方法治疗色素牙的疗效观察(附112例临床分析)

目的 :研究 Opalescence脱色方法对色素牙脱色的疗效。方法 :将 Opalescence脱色剂涂于个别托模唇、颊侧内 ,病人每晚睡前刷牙后擦干牙面 ,戴上托模 ,晨起摘下冲洗干净 ,每天 1次 ,疗程一般为 2周 ,视疗效及患者反应增减用药时间。结果 :1 1 2例患者脱色治疗后氟斑牙的显效率( 80 .8%)高于四环素牙的显效率 ( 66.7%)。有 2 3例病人疗效不甚满意。结论 :应用 Opalescence脱色治疗活髓色素牙效果明显 ,脱色后牙体呈自然色泽 ,透明感好 ,特别适用于年龄较轻、无牙体缺损的氟斑牙及四环素牙的脱色。