心理应激对小鼠脂肪组织黄嘌呤氧化酶表达、活性及相关指标的影响*

目的:探讨心理应激对小鼠脂肪组织黄嘌呤氧化酶表达、活性及相关指标的作用。方法:雄性无特定病原体(SPF)级20只昆明小鼠随机分2组(每组10只),即慢性束缚应激(Stress)组和正常对照(Control)组。Stress组小鼠每天在自制式束缚器中限制活动2 h,其余时间两组小鼠在相同环境中自由饮水摄食,实验持续14 d,取血和白色脂肪组织(WAT);观察脂肪组织病理学改变,检测WAT中黄嘌呤氧化酶(XO)和烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(Nox-4)的蛋白水平,检测WAT组织中XO、Nox-4、超氧化物歧化酶(Mn SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、脂联素(ADPN)、单核细胞趋化蛋白1(MCP-1)、白介素6(IL-6)、肿瘤坏死因子α(TNF-α)、胰岛素受体底物1(IRS-1)、葡萄糖转运蛋白4(GLUT-4)、组织因子(TF)、纤溶酶原激活物抑制物1(PAI-1)的mRNA表达,检测血清和WAT组织中XO酶活性以及血清甘油三酯(TG)、总胆固醇(T-Cho)、游离脂肪酸(FFA)、尿酸(UA)的含量。结果:与control组比较,stress小鼠腹股沟WAT组织中XO免疫染色阳性着色细胞黄褐色沉淀深且丰富,WAT中出现大量的单核细胞、中性粒细胞、嗜酸性粒细胞及浆细胞浸润反应和炎症性的改变;血清XO浓度、WAT组织中XO mRNA水平和XO的酶活性显著升高(P＜0.01),血清游离脂肪酸(FFA)和尿酸(UA)的含量显著增高(P＜0.01),WAT组织中Nox-4蛋白、MCP-1、IL-6、TNF-α、TF、PAI-1mRNA的表达水平显著增高(P＜0.01),而Mn-SOD、GSH-Px、CAT、ADPN、IRS-1和GLUT-4的mRNA水平则显著降低(P＜0.01)。结论:心理应激可诱发脂肪XO过量表达及其活性增高,进而引起脂肪炎症、糖代谢及凝血酶原异常等反应。     

Diagnosis and Surgical Treatment of Renal Hydatid Disease: a Retrospective Analysis of 30 Cases

Echinococcosis (CE) is an infection which is caused by the larval stage of a tapeworm and is endemic in stockbreeding regions of developing countries. The kidney is the most commonly affected organ in the urinary tract. However, reports on renal hydatid disease are limited in the literature, and usually there are no specific clinical characteristics and promising operative methods. The purpose of this study is to assess the most appropriate surgical technique for the patient with urinary tract CE. We retrospectively analyzed thirty patients with renal hydatid cysts who received different surgical treatments in the urology department of the First Affiliated Hospital of Xinjiang Medical University from February 1985 to April 2010. Twenty patients were males and ten were females. The diagnostic accuracy was 74%, 87.5%, and 66.6% respectively by using of ultrasound, CT, and laboratory tests. Thirty patients were followed up for 1–15 years after surgery. One patient experienced a recurrence of renal CE. The ultrasound, CT, and immunological tests are an important means of diagnosis. The surgical treatment principle of renal hydatid should be based on residual renal function, hydatid cyst size, number, location, and surgical techniques to determine the surgical plan to retain the renal function.     

Plasma-free amino acid profiling of cervical cancer and cervical intraepithelial neoplasia patients and its application for early detection

In this study, plasma-free amino acid profiles were used to investigate pre-cancerous cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) metabolic signatures in plasma. Additionally, the diagnostic potential of these profiles was assessed, as well as their ability to provide novel insight into CSCC metabolism and systemic effects. Plasma samples from CIN patients (n = 26), CSCC patients (n = 22), and a control healthy group (n = 35) were analyzed by high-performance liquid chromatography, and their spectral profiles were subjected to the t test for statistical significance. Potential metabolic biomarkers were identified using database comparisons that examine the significance of metabolites. Compared with healthy controls, patients with CIN and CSCC demonstrated lower levels of plasma amino acids; plasma levels of arginine and threonine were increased in CIN patients but were decreased in cervical cancer patients. Additionally, the levels of a larger group of amino acids (aspartate, glutamate, asparagine, serine, glycine, histidine, taurine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine) were gradually reduced from CIN to invasive cancer. These findings suggest that plasma-free amino acid profiling has great potential for improving cancer screening and diagnosis and for understanding disease pathogenesis. Plasma-free amino acid profiles may have the potential be used to determine cancer diagnoses in the early stage from a single blood sample and may enhance our understanding of its mechanisms.     

杏褪绿卷叶植原体新疆分离物核糖体蛋白基因的克隆与序列分析

【目的】了解杏褪绿卷叶植原体新疆分离物的系统发育关系及遗传分化,确定其分类地位。【方法】利用植原体核糖体蛋白(rp)基因的特异性引物rpF1/rpR1对新疆轮台县托克逊县杏褪绿卷叶病植株总DNA进行PCR扩增,并对部分扩增片段克隆、测序及序列分析。【结果】获得杏褪绿卷叶植原体新疆分离物rp基因片段大小为1196 bp,该片段包含部分rpS19以及rpL22和rpS3基因的全部序列。序列相似性和系统进化分析表明,杏褪绿卷叶植原体新疆分离物与16SrⅤ-rp亚组中的各代表性植原体的rp基因核苷酸序列相似性达到95.7%~99.3%,其中与rpⅤ-C亚组的甜樱桃绿化植原体和枣疯病植原体的相似性最高,核苷酸及氨基酸相似性分别达到99.2%~99.3%和98.3%~98.4%。进一步虚拟RFLP分析,发现杏褪绿卷叶植原新疆分离物rp基因的酶切图谱与rpⅤ-C亚组成员相似性最高,但在MseⅠ、SspⅠ和TaqⅠ的酶切位点上存在差异。综上初步判断其可能属于16SrⅤ组(榆树黄化组)中的一个新rp亚组。【结论】本研究首次报道了杏褪绿卷叶植原体新疆分离物的rp基因序列,确定了其分类地位,为杏褪绿卷叶病的早期诊断和检测提供了基础。     

农杆菌介导拒食蛋白XnAFP2转化恢复系多系一号的研究

拒食蛋白是从嗜线虫致病杆菌北京变种CB6菌株分离得到的对多种昆虫具有拒食、抑制生长发育作用的毒性蛋白,通过根癌农杆菌介导转化法将极拒食蛋白基因导入XnAFP2多系一号基因组,获得了转基因水稻植株.通过PCR、RTPCR和Southern blot验证目的基因的整合和表达.     

荷斯坦奶牛中性粒细胞β-防御素cDNA的克隆与序列分析

为研发牛β-防御素生物制剂,防治奶牛乳腺炎。根据已发表的牛β-防御素基因序列设计一对引物。收集患乳腺炎奶牛乳静脉血液中性粒细胞,用试剂盒提取总RNA。经RT-PCR扩增牛β-防御素cDNA片段,回收纯化PCR产物,连接pMD18-T载体,转化感受态细胞JM109。在含X-Gal、Amp及IPTG的LB平板上筛选阳性克隆,经菌落PCR及双酶切鉴定后,对目的片段进行测序。结果表明RT-PCR扩增出的cDNA片段碱基数为189bp,含有一个完整的ORF,能编码63个氨基酸的多肽,多肽中含6个保守半胱氨酸残基。用Blast程序进行检索比较,表明扩增序列与GenBank中发表的BNBD-7编码区序列96.3%相同。结果成功克隆出奶牛β-防御素家族的成员BNBD-7基因,为重组牛β-防御素的开发奠定了基础。     

Decreased expression of EZH2 reactivates RASSF2A by reversal of promoter methylation in breast cancer cells

EZH2, the catalytic subunit of polycomb repressor complex 2, has oncogenic properties, whereas RASSF2A, a Ras association domain family protein, has a tumor suppressor role in many types of human cancer. However, the interrelationship between these two genes remains unclear. Here, we showed that the downregulation of EZH2 reduces CpG island methylation of the RASSF2A promoter, thereby leading to increased RASSF2A expression. Our findings also showed that knockdown of EZH2 increased RASSF2A expression in the human breast cancer cell line MCF‐7 in cooperation with DNMT1. This was similar to the effect of 5‐Aza‐CdR, a DNA methylation inhibitor that reactivates tumor suppressor genes and activated RASSF2A expression in our study. The EZH2 inhibitor DZNep markedly suppressed the proliferation, migration, and invasion of MCF‐7 cells treated with ADR and TAM. EZH2 inhibits the expression of tumor suppressor gene RASSF2A via promoter hypermethylation. Thus, it plays an important role in tumorigenesis and is a potential therapeutic target for the treatment of breast cancer.     

CYP1A1基因单核苷酸多态性与肺癌的相关性研究

目的:探讨代谢酶CYP1A1基因MspI位点多态性与新疆汉族人群肺癌遗传易感性之间的相关性.方法:应用聚合酶链式反应(PCR)-限制性片段长度多态性(RFLP)技术检测59例新疆汉族肺癌和84例新疆汉族健康人的CYP1A1基因MspI位点多态性分布频率,并分析了CYP1A1基因MspI位点多态性与新疆汉族人群肺癌遗传易感性和患者性别之间的相关性.结果:(1)CYP1A1基因MspI位点3种多态基因型分布频率在两组间比较差异有统计学意义(χ2=6.682,P=0.035),CC基因型在病例组的分布频率显著高于正常对照组.(2)携带突变CC基因型的个体较携带TT基因型的个体患肺癌的危险性增加(OR=3.759.95%CI=1.228-11.494,P=0.035).(3)男女肺癌患者的CYP1A1基因MspI位点基因型及等位基因频率的差异均无显著性(P>0.05).结论:(1)CC突变基因型可能是新疆汉族人群的肺癌易感因素.(2)CYP1A1基因MspI位点多态性可能与新疆汉族肺癌患者的性别无关.     

Indoxyl Sulfate-Induced Activation of (Pro)renin Receptor Promotes Cell Proliferation and Tissue Factor Expression in Vascular Smooth Muscle Cells

Chronic kidney disease (CKD) is associated with an increased risk of cardiovascular disease (CVD). (Pro)renin receptor (PRR) is activated in the kidney of CKD. The present study aimed to determine the role of indoxyl sulfate (IS), a uremic toxin, in PRR activation in rat aorta and human aortic smooth muscle cells (HASMCs). We examined the expression of PRR and renin/prorenin in rat aorta using immunohistochemistry. Both CKD rats and IS-administrated rats showed elevated expression of PRR and renin/prorenin in aorta compared with normal rats. IS upregulated the expression of PRR and prorenin in HASMCs. N-acetylcysteine, an antioxidant, and diphenyleneiodonium, an inhibitor of nicotinamide adenine dinucleotide phosphate oxidase, suppressed IS-induced expression of PRR and prorenin in HASMCs. Knock down of organic anion transporter 3 (OAT3), aryl hydrocarbon receptor (AhR) and nuclear factor-κB p65 (NF-κB p65) with small interfering RNAs inhibited IS-induced expression of PRR and prorenin in HASMCs. Knock down of PRR inhibited cell proliferation and tissue factor expression induced by not only prorenin but also IS in HASMCs.

Conclusion

IS stimulates aortic expression of PRR and renin/prorenin through OAT3-mediated uptake, production of reactive oxygen species, and activation of AhR and NF-κB p65 in vascular smooth muscle cells. IS-induced activation of PRR promotes cell proliferation and tissue factor expression in vascular smooth muscle cells.     

南疆棉区棉花品种的特性演变与育种潜力研究

南疆棉区经过5次大的品种更换,高产品种的选育起了非常重要的作用,新育成的品种比早期育成的品种产量以每年12.1kg/hm2的速度增长,品种改良对产量的贡献率为43.9%。衣分、单株结铃数和单铃重的总的演变趋势是增加的,农艺性状方面株高降底,株宽增加,自育品种的现蕾量较低,脱落率较高,南疆棉花品种的综合品质变化不大,特别是棉花生产品质未发生根本性变化。南疆棉花品种在单株结铃数、衣分、增加现蕾数、降低花蕾脱落、株宽、果枝始节、高光合速率品种的选育以及纤维比强度等方面具备较大的选择潜力。