Hypothalamic Syndrome and Central Sleep Apnoea Associated with Toluene Exposure

We describe a young man who developed extensive hypothalamicdysfunction including diabetes insipidus, adipsia, hyperprolactinaemia,and poikiliothermia together with central sleep apnoea followingexposure to toluene.     

小鼠Nanog基因的克隆及对人宫颈癌上皮细胞的作用

目的:克隆小鼠Nanog基因并构建带绿色荧光蛋白的真核表达载体pG-Nanog,观察其对人宫颈癌上皮细胞(Hela细胞)中的表达,旨在为进一步观察其对成体细胞的表型变化及细胞增殖奠定前期实验学基础。方法:实验于2006-03/09在西北农林科技大学陕西省干细胞研究中心完成。Nanog基因的克隆参照庄淑珍的方法。Nanog基因真核表达载体的构建参照GeneBank中的小鼠Nanog基因序列,以pNA992为模板扩增Nanog基因,PCR产物以BglⅡ和SacⅡ双酶切,同时将pEGFP-C1用BglⅡ和SacⅡ鉴定,将该质粒命名为pG-Nanog。Hela细胞用含10%新生牛血清的Dulbecco’s改良培养基(Dulbecco’sModifiedEagleMedium,DMEM)培养,转染Hela细胞。转染前1d,在6孔板的每个孔中接种1.2×105个细胞,待细胞生长至60%～70%汇合时,取pG-Nanog与空载体各4μg分别加入500μL无血清无抗生素的DMEM培养液中,同时将6μL稀释于500μL无血清无抗生素的DMEM(干粉)培养液中,将两者混合,室温静置20min,将复合物加入到细胞中,置37℃,体积分数0.05的CO2培养箱中转染24h后吸出复合物加入完全培养基,48h后观察荧光。合成内源对照β-actin,收集转染4d后的细胞提取RNA,反转录为cDNA,然后分别扩增Nanog基因和β-actin基因。采用RT-PCR的方法检测Hela细胞中Nanog基因的表达。结果:①pEGFP-C1载体经双酶切后获得约1kbp的Nanog基因真核表达载体片段,同预期结果相一致,测序结果同GeneBank中的序列同源性达到99.7%。②将转染48h后的Hela细胞置于荧光显微镜下观察,可见明显的荧光,转染pG-Nanog的细胞绿色荧光蛋白集中于细胞核,将转染4d后Hela细胞的总RNA进行RT-PCR检测,产物经琼脂糖电泳分析,只有转染pG-Nanog的细胞中才能够检测到Nanog基因的相应条带。③转染48h后,对细胞进行抗增殖细胞核抗原免疫组化染色,未转染细胞和转染空质粒细胞及转染pG-Nanog细胞染色结果均呈阳性,转染了pG-Nanog的Hela细胞与正常细胞和转染空载体的细胞相比细胞形态发生了一定的改变,细胞表面形成了许多突起。结论:小鼠Nanog基因的克隆、真核表达载体的构建及在Hela细胞中的表达均获得成功,并观察到Hela细胞发生了形态的改变。     

Effects of the temperature, the duration of frozen storage, and the freezing container on in vitro measurements in human peripheral blood mononuclear cells

Background: Bone marrow, peripheral blood, and umbilical cord blood have been used to prepare autologous and allogeneic pluripotential mononuclear cells for use in the repopulation of bone marrow. Study Design and Methods: The purpose of this study was to evaluate how the temperature and duration of frozen storage of human peripheral blood mononuclear cells (PBMCs), as well as the freezing container, affected the in vitro recovery and viability of the mononuclear cells and their growth in colony-forming unit-granulocytic-erythroid-monocytic- megakaryocytic (CFU-GEMM) tissue culture assay. PBMCs were isolated from ficoll-hypaque-treated cellular residue obtained during the plateletpheresis of blood from 15 healthy donors. The PBMCs were treated with dimethyl sulfoxide (DMSO) to achieve a final DMSO concentration of 10 percent. Each unit was then separated into six aliquots: one stored in a polyvinylchloride (PVC) plastic bag, one in a polyolefin plastic bag, and four in polyethylene cryostorage vials. Each aliquot was frozen in a -80 degrees C mechanical freezer at a freezing rate of 2 to 4 degrees C per minute. The frozen PBMCs in PVC bags were stored in a -80 degrees C mechanical freezer and those in polyolefin bags in a -135 degrees C mechanical freezer. Each of the four frozen samples in a vial was stored at a different temperature: one in the -80 degrees C freezer, one in the -135 degrees C freezer, one in the vapor phase of liquid nitrogen at -150 degrees C, and one in liquid nitrogen at -197 degrees C. Some of the frozen PBMCs were stored for periods of 1 to 1.5 years and others for 2 to 2.4 years, after which they were thawed, washed, and tested. Results: The samples stored in PVC bags and those stored in polyolefin bags exhibited in vitro recoveries that were 90 percent of the recovery of fresh PBMCs and viabilities of 90 percent after 2.4 years of frozen storage. The PBMCs stored in PVC bags exhibited no loss of CFU-GEMM activity after 1 to 1.5 years, but a 40-percent loss of activity was observed after 2 to 2.4 years. PBMCs stored in polyolefin bags, however, exhibited no loss of CFU-GEMM activity, even after 2 to 2.4 years of storage. In vitro recovery was significantly lower in PBMCs stored in vials at -80 degrees C or -135 degrees C than in cells stored in PVC or polyolefin bags at these temperatures, both in the 1- to 1.5-year and the 2- to 2.4-year time frames. In vitro recovery and viability were similar in PBMCs stored in vials at -80 degrees C, -135 degrees C, -150 degrees C, and -197 degrees C. The growth patterns in the CFU-GEMM assay in PBMCs stored in vials were significantly lower after storage at -80 degrees C than after storage at -135 degrees C, -150 degrees C, or -197 degrees C. Conclusion: PBMCs isolated by leukapheresis and ficoll-hypaque treatment can be frozen with 10-percent DMSO in a -80 degrees C mechanical freezer. When a PVC bag is used for freezing and storage of PBMCs at -80 degrees C, the duration of frozen storage should not exceed 1.5 years, whereas PBMCs frozen in a polyolefin bag can be stored in a -135 degrees C freezer for as long as 2.4 years. When these guidelines were followed, in vitro recovery was 90 percent that of fresh PBMCs, viability was 90 percent, and growth in the CFU-GEMM tissue culture assay was similar to that of fresh PBMCs. The PBMCs frozen and stored in PVC or polyolefin bags exhibited satisfactory results, whereas those stored in cryostorage vials did not.     

Should Hepatic Metastatic Colorectal Cancer Patients with Extrahepatic Disease Undergo Liver Resection/Ablation?

Background  

Surgical therapy has been proven to be the mainstay of treatment for hepatic metastases from colorectal cancer (CRM) in the appropriate patient. Previous contraindications were patients with extrahepatic disease (EHD) do not benefit from liver resection or ablation. We hypothesized that the survival of patients with EHD who receive aggressive multimodality care would be the same as those without EHD.     

Epithelial NF-kappaB activation promotes urethane-induced lung carcinogenesis

Chronic inflammation is linked to carcinogenesis in several organ systems. In the lungs, NF-kappaB, a central effector of inflammatory responses, is frequently activated in non-small-cell lung cancer, but its role in tumor promotion has not been studied. Several lines of evidence indicate that ethyl carbamate (urethane)-induced lung tumor formation, a prototypical mouse model of multistage lung carcinogenesis, is potentiated by inflammation. We found that mouse strains susceptible to lung tumor formation (FVB, BALB/c) exhibited early NF-kappaB activation and inflammation in the lungs after urethane treatment. However, a resistant strain (C57B6) failed to activate NF-kappaB or induce lung inflammation. In FVB mice, we identified urethane-induced NF-kappaB activation in airway epithelium, as well as type II alveolar epithelial cells and macrophages. Using an inducible transgenic mouse model (FVB strain) to express a dominant inhibitor of NF-kappaB specifically in airway epithelial cells, we found that urethane-induced lung inflammation was blocked and tumor formation was reduced by >50%. Selective NF-kappaB inhibition resulted in increased apoptosis of airway epithelial cells at 2 weeks after urethane treatment in association with a marked reduction of Bcl-2 expression. These studies indicate that NF-kappaB signaling in airway epithelium is integral to tumorigenesis in the urethane model and identify the NF-kappaB pathway as a potential target for chemoprevention of lung cancer.     

Retroperitoneal and visceral sarcomas: issues for the general surgeon

Sarcomas represent a rare, heterogeneous group of malignant tumors that arise from the mesenchymal tissues of the body. Although infrequently encountered, these tumors generate vigorous academic interest and an ever-expanding volume of medical literature. Chemotherapy is widely regarded as ineffective because of the often-large tumor burden and lack of good therapeutic drugs. Radiation therapy is often difficult to administer because of locoregional toxicity. Fortunately, targeted immunologic therapies have shown promise in some specific gastrointestinal mesenchymal tumors. To date, sarcoma remains a malignancy best treated operatively. Given the wide heterogeneity and biology of these tumors and the amount of new data available, a review of the current literature is warranted. The first installment of this review series dealt with extremity and trunk soft tissue sarcomas; this one will focus on retroperitoneal and visceral sarcomas and the management challenges they pose.     

Kentucky hepatoma: epidemiologic variant or same problem in a different region?

BACKGROUND: Hepatitis and cirrhosis are common etiologic factors in hepatocellular carcinoma (HCC) in the United States. However, noncirrhotic, nonfibrotic HCC has been recognized more frequently in Kentucky. The aim of this study was to evaluate the epidemiologic features of this variant of HCC. HYPOTHESIS: Kentucky hepatoma, defined as a noncirrhotic, nonfibrotic, hepatitis-negative HCC, occurs in an older population with more favorable preoperative factors when compared with other patients with HCC. DESIGN: A prospective review of our 1002 hepatopancreaticobiliary patients, the Kentucky Cancer Registry, and the Surveillance, Epidemiology, and End Results database. SETTING: An academic referral center. PATIENTS: All patients with HCC treated from January 1, 1999, through September 30, 2005, were reviewed for clinicopathologic factors, recurrence, and outcome. RESULTS: In a review of the region's 703 patients with HCC, we have seen a 4-fold increase in age-specific HCC diagnosis, with the most rapid increase seen in the 60- to 69-year-old age group. In our institution's 103 patients with HCC, 62 (60.2%) were without hepatitis or cirrhosis. These noncirrhotic, hepatitis-free patients were found to be significantly older (70 vs 55 years; P = .001), to be more often female (40.3% vs 24.4%; P = .01), to have a larger tumor size (6.5 vs 3.9 cm; P = .004), to have fewer liver lesions (1 vs 3; P = .22), and to more frequently undergo surgical therapy (75.6% vs 53.8%; P = .01) than the patients with cirrhosis or hepatitis (n = 41). CONCLUSIONS: A larger percentage of the patients with HCC seen in our region are significantly different from those in other reports throughout the United States in preoperative clinical and pathologic presentation. The reason for this change is as yet unknown, but the incidence continues to rise annually.     

Review of the Lynch syndrome: history, molecular genetics, screening, differential diagnosis, and medicolegal ramifications

Platelets have a central role in the development of arterial thrombosis and subsequent cardiovascular events. An appreciation of this complex process has made antiplatelet therapy the cornerstone of cardiovascular disease management. However, numerous patients will experience a recurrent atherothrombotic vascular event despite adequate antiplatelet therapy. Individual differences in the rate of platelet activation and reactivity markedly influence normal hemostasis and the pathological outcome of thrombosis. Such an individual variability is largely determined by environmental and genetic factors. These are known to either hamper platelets' response to agonists, and thereby mimic the pharmacological modulation of platelet function or mask therapy effect and sensitize platelets. In this article, we reviewed the antiplatelet mechanisms of aspirin and clopidogrel and the possible role of different polymorphisms, which may affect the efficacy of antiplatelet therapy. Heterogeneity in the way patients respond to aspirin and clopidogrel may in part reflect variation in cyclooxygenase (COX)-1, COX-2, glycoprotein (GP) Ib alpha, GP Ia/IIa, GP IIb/IIIa, UGT1A6*2, P2Y₁, P2Y₁₂, CYP2C9, CYP3A4 and CYP3A5 genotypes.