马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

为了解新疆马疱疹病毒1型（EHV-1）主要毒力基因遗传进化情况并构建TK基因缺失株，本研究以EHV-1 XJ2015株DNA为模板，对其主要毒力基因TK、gI和gE全长进行克隆、测序及生物信息学分析，并扩增TK基因左右重组臂TKL和TKR，构建质粒pUC-TKLR，将扩增后的增强绿色荧光蛋白（EGFP，含有CMV+polyA）插入pUC-TKLR质粒，构建TK基因缺失打靶质粒。TK、gI和gE基因同源性分析结果显示，XJ2015株与国外EHV-1分离株TK、gI和gE基因同源性均较高，分别为99.8%~100.0%、99.6%~100.0%和99.9%~100.0%；与EHV-3分离株同源性均最低，分别为72.9%、59.4%和62.1%；遗传进化分析显示，3个基因均与国外EHV-1同属于一个遗传进化分支，与EHV-9和EHV-4进化关系较近，但与EHV-3进化关系较远，表明XJ2015毒株与国外EHV-1毒株TK、gI、gE基因核苷酸上差异不明显，没有明显的地域性特征，功能基因保守且进化缓慢，同源基因功能相同或相近；经PCR扩增、酶切、测序及转染鉴定，本试验成功构建了用于TK基因缺失的打靶质粒pUC-TKLR-EGFP。通过对EHV-1主要毒力基因的分析及TK基因缺失打靶载体的构建，为新疆地区马鼻肺炎流行病学调查分析、TK基因缺失株的构建提供理论依据。     

猎豹与虎猫杯状病毒的分离及其超变区基因比较研究

用F81细胞从上海某动物园患口腔溃疡的猎豹和虎的唾液病料中分离获得两株杯状病毒，经形态学、理化学、生物学鉴定和病毒核酸超变区基因RT-PCR扩增与序列测定证明两株病毒均为猫杯状病毒(FCV)，分别命名为FCV/cheetah/Shanghai/02/2002与FCV／tiger/Shanghai／03/2002。人工感染猫可引起体温升高，口腔溃疡，食欲下降等症状。两株病毒的超变区序列520bp长片段问的同源性为99.2％，与国内桂林虎分离株(TFCV9710)的同源性为74．0％，与国外分离株的总体同源性为58．1％，说明不同宿主或同种不同个体间杯状病毒分离株超变区核酸差异十分显著，符合猫杯状病毒的分子生物学特征。     

ULTRASONOGRAPHY OF ALIMENTARY LYMPHOSARCOMA IN THE CAT

Clinical, radiological and ultrasonographic findings from 22 cats with gastrointestinal lymphosarcoma were reviewed. The most common clinical findings were anorexia, weight loss, vomiting, lethargy, depression, fever, anemia and a palpable abdominal mass. Abdominal radiographs of 12 cats revealed a mass associated with the gastrointestinal tract and/or mesentery, and decreased serosal detail, suggestive of peritoneal effusion. The most common ultrasonographic finding was transmural gastric or intestinal thickening associated with loss of normal wall layering, reduced wall echogenicity and localized hypomotility. As is noted in people, other patterns such as transmural-segmental, transmural-nodular, transmural-bulky and mucosal infiltration were represented. In 3 cats, mesenteric lymph-adenopathy was responsible for most of the mass-effect. Fine-needle aspiration biopsy, automated microcore biopsy, endoscopic biopsy or necropsy confirmed the diagnosis of lymphosarcoma in all cats. Chemotherapy was instituted in 19 cats. Only 6 cats had a complete response to treatment. The results of this study indicate that ultrasonography is a valuable tool for the diagnosis of feline alimentary lymphosarcoma and that biopsies can be obtained under ultrasonographic guidance.     

Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

牛磺酸在猫营养中的应用

主要综述近年猫的牛磺酸营养,从研究方法来看衡量牛磺酸利用的方法与指标已较完善,对牛磺酸的代谢有了深入的了解,并进一步关注猫肠道菌群影响牛磺酸代谢的机理的研究;初步揭示大豆蛋白和脂肪等营养物质降低牛磺酸利用率的原因;在猫粮加工方面指出不良热处理导致的猫粮中牛磺酸利用率降低的机制,并分析了一些不适合配制猫粮的原料如米糠、大豆蛋白对牛磺酸代谢的影响;以及在猫粮中直接添加晶体牛磺酸的重要性。从文中可以看到,作为必需氨基酸的牛磺酸对于猫的生长繁殖来说十分重要。     

A review of feline serum protein electrophoresis

A review of the current literature available on feline serum proteins is presented. Early studies concentrated on comparative aspects of species variations in the electrophoretic pattern. The feline electrophoretogram was divided into five basic regions: albumin, alpha-1, alpha-2, beta and gamma. Different subdivisions of these areas were recognized depending on the support medium used. Current papers have compared the relative migration distances of each globulin peak to the migration of albumin. This "Rf" value enables reliable peak identification. To date, no data exists identifying the individual proteins responsible for the peaks in the alpha and beta regions. The only feline globulin to be studied is haptolobin; however its precise location on the electrophoretic strip was not identified.     

Chylothorax associated with cryptococcal mediastinal granuloma in a cat

A 10-year-old neutered female cat had chylothorax, precaval syndrome, and a mediastinal granuloma resulting from infection with Cryptococcus neoformans. Diagnosis of a chylous effusion was made by cytologic examination of pleural fluid and by finding higher triglyceride levels in the effusion than in serum (825 vs. 64 mg/dl, respectively). Postmortem examination revealed cryptococcal organisms in the mediastinal granuloma, lungs, cerebral meninges, and connective tissues adjacent to the thyroid gland. Chylous effusion in a cat associated with cryptococcosis has not been reported previously. Cryptococcosis should be included in the differential diagnosis in chylous effusions in cats.     

Lipoprotein electrophoresis differentiation of chylous and nonchylous pleural effusions in dogs and cats and its correlation with pleural effusion triglyceride concentration

Pleural effusions from 23 dogs and 25 cats hospitalized between November 1986 and April 1987 at the Veterinary Hospital of the University of Pennsylvania (VHUP) were classified as chylous (chylomicrons present) or nonchylous (chylomicrons absent) by the presence or absence, respectively, of a chylomicron band on lipoprotein electrophoresis gels of the effusions. Triglyceride concentrations, cholesterol concentrations, and cholesterol/triglyceride (C/T) ratios were compared between the chylous and nonchylous groups in each species. Cholesterol concentrations were not significantly different between chylous and nonchylous effusions in both dogs and cats. Cholesterol/triglyceride ratios of less than 1 were present in all dogs and cats with chylous effusions; however, 12% of dogs with nonchylous effusions and 50% of cats with nonchylous effusions also had a C/T ratio less than 1. Triglyceride concentrations accurately classified all effusions as chylous or nonchylous in both dogs and cats. Pleural effusions with triglyceride concentrations greater than l00 mg/dl were chylous in all cases, whereas, effusions with triglyceride concentrations less than l00 mg/dl were nonchylous in all cases.     

Iatrogenic magnesium overdose: 2 case reports

Objective: To describe the clinical manifestations and treatment of hypermagnesemia and the potential drug errors that can lead to iatrogenic electrolyte toxicities. Summary: We report 2 cases of iatrogenic intravenous (IV) magnesium (Mg) overdose. Both cases developed extreme cardiovascular and neurologic symptoms consisting of vomiting, hypotension, bradycardia, flaccid paralysis, and severe mental depression. Diagnosis was made based upon serum ionized Mg levels (3.47 mmol/L; reference range: 0.43–0.58 mmol/L for Case #1; and 4.64 mmol/L; reference range: 0.42–0.55 mmol/L for Case #2). Each animal was treated with 0.9% NaCl for diuresis and IV calcium gluconate. Within 24 hours, the cardiovascular and neurologic status of both animals, as well as the serum Mg concentration, had normalized. Each animal was discharged with no complications. Both animals had been hospitalized for critical illness and had developed hypomagnesemia that was being treated with Mg sulfate infusions. The cause for the hypermagnesemia was due to miscalculations in treatment orders that led to erroneously administered Mg‐containing solutions. Confusing drug labels and varying units of measurement can lead to erroneous miscalculations, especially in critically ill patients that receive multiple IV infusions. New information provided: This is the first case report of iatrogenic Mg overdose in veterinary medicine. These 2 cases had a good clinical outcome with prompt recognition and supportive care.     

Effects of oral orbifloxacin on fecal coliforms in healthy cats: a pilot study

The study objective was to determine the effect of oral orbifloxacin (ORB) on antimicrobial susceptibility and composition of fecal coliforms in cats. Nine cats were randomized to two groups administered a daily oral dose of 2.5 and 5.0 mg ORB/kg for 7 days and a control group (three cats per group). Coliforms were isolated from stool samples and were tested for susceptibilities to ORB and 5 other drugs. ORB concentration in feces was measured using high-performance liquid chromatography (HPLC). The coliforms were undetectable after 2 days of ORB administration, and their number increased in most cats after termination of the administration. Furthermore, only isolates of Escherichia coli were detected in all cats before administration, and those of Citrobacter freundii were detected after termination of the administration. E. coli isolates exhibited high ORB susceptibility [Minimum inhibitory concentration (MIC), ≤0.125 µg/ml] or relatively low susceptibility (MIC, 1−2 µg/ml) with a single gyrA mutation. C. freundii isolates largely exhibited intermediate ORB susceptibility (MIC, 4 µg/ml), in addition to resistance to ampicillin and cefazolin, and harbored qnrB, but not a gyrA mutation. HPLC revealed that the peaks of mean concentration were 61.3 and 141.0 µg/g in groups receiving 2.5 and 5.0 mg/kg, respectively. Our findings suggest that oral ORB may alter the total counts and composition of fecal coliform, but is unlikely to yield highly fluoroquinolone-resistant mutants of E. coli and C. freundii in cats, possibly because of the high drug concentration in feces.