Prevalence of ophthalmic disease in blue‐eyed horses

A perception exists amongst referring veterinarians and the lay public that blue‐eyed horses have increased frequency of ocular disease. The aim of this retrospective study was to assess the prevalence of ocular disease in horses with blue or heterochromic eyes relative to those with brown eyes. The medical records of horses presenting to either the Comparative Ophthalmology services or Equine Medicine/Surgery services at 2 institutions were reviewed. Signalment, ocular and nonocular diagnoses were recorded. Ocular disease was divided into 4 categories: adnexa, cornea, intraocular/orbit and squamous cell carcinoma (SCC). Owners were contacted by telephone to confirm iris colour. Chi‐square analysis was used to compare group proportions. A total of 164 eyes of horses with ocular disease and 212 eyes of horses without ocular disease were included. Blue eyes were equally common in the ocular disease and nonocular disease groups (P = 0.265). There was no significant difference in the proportion of blue‐ and brown‐eyed horses when comparing the adnexal and corneal (P = 0.548), corneal and intraocular/orbit (P = 0.379) and adnexal and intraocular/orbit (P = 0.843) categories. A significant difference was detected in the proportion of blue‐eyed horses between the adnexal (P = 0.000), corneal (P = 0.033), intraocular/orbital (P = 0.000) and SCC categories, with a higher proportion of blue‐eyed horses in the SCC than in the other 3 groups. Horses with blue or heterochromic irides are more likely to develop ocular SCC than horses with brown irides, but are not more likely to have adnexal, corneal or intraocular/orbital disease or to be presented for evaluation of ophthalmic disease. Veterinarians should consider the results of this study when answering questions from the lay public regarding the predisposition of blue‐eyed horses to ocular disease.     

Effect of Oversulfation on the Composition,Structure, and In Vitro Anti-Lung Cancer Activity of Fucoidans Extracted from Sargassum aquifolium

Intensive efforts have been undertaken in the fields of prevention, diagnosis, and therapy of lung cancer. Fucoidans exhibit a wide range of biological activities, which are dependent on the degree of sulfation, sulfation pattern, glycosidic branches, and molecular weight of fucoidan. The determination of oversulfation of fucoidan and its effect on anti-lung cancer activity and related signaling cascades is challenging. In this investigation, we used a previously developed fucoidan (SCA), which served as a native fucoidan, to generate two oversulfated fucoidan derivatives (SCA-S1 and SCA-S2). SCA, SCA-S1, and SCA-S2 showed differences in compositions and had the characteristic structural features of fucoidan by Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) analyses. The anticancer properties of SCA, SCA-S1, and SCA-S2 against human lung carcinoma A-549 cells were analyzed in terms of cytotoxicity, cell cycle, Bcl-2 expression, mitochondrial membrane potential (MMP), expression of caspase-3, cytochrome c release, Annexin V/propidium iodide (PI) staining, DNA fragmentation, and the underlying signaling cascades. Our findings indicate that the oversulfation of fucoidan promotes apoptosis of lung cancer cells and the mechanism may involve the Akt/mTOR/S6 pathway. Further in vivo research is needed to establish the precise mechanism whereby oversulfated fucoidan mitigates the progression of lung cancer.     

p27基因在宫颈癌组织中的表达及其意义

目的：探讨肿瘤抑制基因p27在宫颈癌组织中的表达及其临床意义。方法：应用免疫组织化学方法（SP法）检测p27基因在62例官颈癌组织和20例正常官颈组织中的表达。结果：62例宫颈癌组织中,p27的阳性表达率为38．7％（24／62）;p27的阳性表达与宫颈病灶大小、临床分期、细胞分化程度、转移及预后有密切关系（P〈0．01或〈0．05）;与宫颈癌的病理类型及年龄无关（P〉0．05）。结论：p27基因的表达与宫颈癌的生长、浸润及淋巴结转移有关,其表达水平的高低可作为判断宫颈癌预后的重要指标之一。     

双歧杆菌完整肽聚糖的体外抗肿瘤作用及对大鼠大肠肿瘤的影响

目的：观察双歧杆菌完整肽聚糖（WPG）体外抗肿瘤作用及对大鼠大肠肿瘤的影响。方法：采用MTT法检测双歧杆菌WPG对人结肠癌细胞株LS-174T、人胃粘膜上皮细胞株GES-1的杀伤作用。以二甲肼（DMH）为诱发荆,观察双歧杆菌WPG对大鼠大肠肿瘤的防治作用;DMH诱癌及双歧杆菌WPG灌胃时间为18周,在第33周结束实验。结果：双歧杆菌WPG在体外对人结肠癌细胞株具有显著的选择性杀伤作用,体内给药可明显降低大鼠大肠肿瘤发生率（60％vs100％,P〈0．01）．但大肠癌平均个数及肿瘤体积指数的差异无显著性（P〉0．05）。结论：双歧杆菌的WPG对DMH诱导的大鼠大肠肿瘤具有预防作用。     

应用抑制消减杂交技术克隆鼻咽癌转移调控基因

目的：分离并鉴定高低转移表型不同的人鼻咽癌细胞株差异表达cDNA序列,了解鼻咽癌转移抑制基因或具有抑制活性的核苷酸序列。方法：以具有高低转移潜能不同的人鼻咽癌细胞株为研究对象,应用抑制消减杂交技术（suppression subtractive hybridization,SSH）和T／A克隆法构建差异表达cDNA消减文库,对差异表达的cDNA进行测序和生物信息学分析。结果：成功构建人鼻咽癌细胞克隆株差异表达cDNA消减文库,从随机测序10个克隆中获得6个在低转移鼻咽癌细胞株中高表达cDNA序列,它们都与已知的人类基因片段具有高度同源性。结论：应用抑制消减杂交技术,从一对具有高低转移表型差异的鼻咽癌细胞株中获得6条可能与肿瘤转移抑制相关的cDNA序列,它们可能在维持肿瘤细胞的稳定和抑制肿瘤转移中起重要作用。     

肝癌细胞系中Runx3基因表达及启动子区异常甲基化分析

目的：通过检测6种肝癌细胞中Runx3基因异常甲基化状态及表达情况,探讨药物5-aza-2＇-deoxycytidine（decitabine）激活Runx3基因重新表达的能力及对肝癌细胞生长的影响。方法：采用DNA甲基化特异性PCR（MSP）技术分别对6种肝细胞癌细胞系Runx3基因启动子区域甲基化状态进行检测,同时采用逆转录一聚合酶链反应（RT—PCR）检测5种肝癌细胞中Runx3的表达情况及药物5-aza-2＇-deoxycytidine处理其中3种肝癌细胞前后Runx3表达变化。结果：6种肝癌细胞系中,有3种细胞Runx3基因启动子区域存在甲基化异常,药物5-aza-2＇-deoxycytidine处理3种肝癌细胞后,Runx3基因明显表达或表达活性增强。结论：启动子区异常甲基化是导致Runx3基因失活的主要原因之一,与肝细胞癌发生早期密切相关,可作为肝细胞癌早期诊断的分子标记物及分子治疗的靶点。     

人乳头瘤病毒16L1蛋白的原核表达及免疫原性研究

采用聚合酶链反应技术从宫颈癌组织中获得人乳头瘤病毒(HPV)16L1基因,克隆到pGEM—TEasy载体中,再连接到原核表达载体pGEX-6p-1谷胱甘肽-S-转移酶基因的下游,重组质粒转化大肠杆菌BL21,经IPTG诱导、SDS—PAGE分析,用表达产物免疫小鼠,获得的血清进行间接免疫荧光检测。结果表明：获得的pGEX-6p-1-L1基因在大肠杆菌中能以谷胱甘肽-S-转移酶融合蛋白的形式得到高效表达,表达产物免疫小鼠,间接免疫荧光显示免疫血清能与peDNA—L1转染的B16细胞呈现特异反应,说明体外表达的HPV16L1蛋白保留了天然蛋白的抗原性,可作为HPV的诊断抗原、免疫学分析和疫苗研制的原料。