Detection of Clinically Relevant Pain Relief in Cats with Degenerative Joint Disease Associated Pain

Background

Detection of clinically relevant pain relief in cats with degenerative joint disease (DJD) is complicated by a lack of validated outcome measures and a placebo effect.

Hypothesis/Objectives

To evaluate a novel approach for detection of pain relief in cats with DJD.

Animals

Fifty‐eight client‐owned cats.

Methods

Prospective, double‐masked, placebo‐controlled, stratified, randomized, clinical study. Enrolled cats were 6–21 years of age, with owner‐observed mobility impairment, evidence of pain in at least 2 joints during orthopedic examination, and overlapping radiographic evidence of DJD, and underwent a 2‐week baseline period, 3‐week treatment period with placebo or meloxicam, and 3‐week masked washout period. Outcome measures were evaluated at days 0, 15, 36, and 57.

Results

Both groups significantly improved after the treatment period (day 36) on client‐specific outcome measures (CSOM) and feline musculoskeletal pain index (FMPI) (P < .0001 for both); there was no difference between the groups on CSOM or FMPI score improvement. After the masked washout period, more cats that received meloxicam during the treatment period had a clinically relevant decrease in CSOM score (P = .048) and FMPI score (P = .021) than cats that received placebo.

Conclusions and Clinical Importance

Using both a client‐specific and a general clinical metrology instrument, owners of cats with DJD were able to detect evident recurrence of clinical signs after withdrawal of active medication than after withdrawal of placebo, and that this study design might be a novel and useful way to circumvent the placebo effect and detect the efficacy of pain‐relieving medications.     

Effect of dietary supplementation with an ethanolic extract of propolis on broiler intestinal morphology and digestive enzyme activity

The present study aimed to evaluate the effect of different levels of an ethanolic extract of propolis (EEP) on broiler performance, carcass characteristics, weight of gastrointestinal organs, intestinal morphometry and digestive enzyme activity. 1020 male broiler chicks were assigned in a completely randomised experimental design to six treatments (EEP supplement levels of 0, 1000, 2000, 3000, 4000 and 5000 ppm) and five replications, and 34 birds per experimental unit. The experimental diets were administered from 1 to 21 days of age, and the birds were subsequently provided a ration based on corn and soybean meal. EEP supplementation from 1 to 7 days negatively affected (p < 0.05) the weight gain and feed intake. The proventriculus weight at 7 days exhibited a quadratic response (p < 0.05), which predicted a lower weight at a dose of 2865 ppm of the EEP. For the duodenum at 21 days of age, the response pattern (p < 0.05) predicted that birds that were fed 2943 and 3047 ppm of the EEP would exhibit an improved crypt depth and villus‐to‐crypt ratio respectively. The villus height, crypt depth and villus‐to‐crypt ratio in the jejunum and the ileum were not affected (p > 0.05). With increased EEP doses, the duodenal sucrase activity linearly decreased at 7 days of age and linearly increased in the jejunum at 21 days of age (p < 0.05), while pancreatic enzyme activity was unaffected (p > 0.05). Although the carcass and cut yields did not improve, the percentage of abdominal fat decreased (p < 0.05). The supplementation of the broiler pre‐starter diet with 1000–5000 ppm of the EEP impaired performance at this stage, most likely due to the decreased sucrase activity. However, the EEP supplementation from 3000 ppm improved intestinal morphophysiology at 21 days of age and did not affect the performance or carcass yield at 42 days of age.     

水牛RNA聚合酶Ⅲ启动子的克隆与鉴定

[目的]获得有效的水牛RNA聚合酶Ⅲ启动子序列,为开展水牛源细胞的基因特异沉默研究奠定基础.[方法]通过启动子上、下游保守序列对水牛源启动子7SK、U6进行克隆和启动子关键顺式作用元件识别,利用一段针对EGFP的shRNA片段（shEGFP）对水牛7SK、U6启动子进行功能性分析,然后分别在水牛源及鼠源细胞中与pEGFP-N1共转染,转染48h后用荧光显微镜检测EGFP的表达情况,并用流式细胞仪和荧光实时定量PCR检测EGFP沉默表达情况.[结果]克隆获得水牛7SK、U6启动子序列分别为430和357 bp,其OCT-1（或CACCC盒）及TATA盒高度保守.连接shEGFP后与pEGFP-N1共转染细胞,通过荧光显微镜可观察到细胞发生了明显的荧光表达沉默现象;通过流式细胞分析,发现水牛7SK和U6启动子引导的shEGFP在水牛源细胞中沉默效率高达93.82％和87.45％;荧光实时定量PCR检测结果显示,在转染bu7SK-shEGFP和buU6-shEGFP的水牛源BFF细胞中,EGFP表达水平均显著低于其他物种启动子的细胞转染组（P＜0.05）,而在鼠源PT67细胞中,水牛启动子的启动效率与其他物种启动子差异不显著（P＞0.05）.[结论]水牛7SK和U6启动子可高效启动shRNA表达,且具有一定的物种特异性.     

甘蓝型油菜WRI1基因cDNA克隆与种子特异性表达载体的构建

WRI1基因所编码的转录因子参与糖酵解和脂肪酸合成,在种子油的积累过程中起着关键作用。在种子中超表达WRI1基因,对提高油菜种子含油量具有重要意义。本研究利用RT-PCR技术,从甘蓝型油菜湘油15号cDNA中分离克隆了5个WRI1基因全长cDNA序列,选取2种存在明显差异(碱基缺失)的cDNA序列构建植物种子特异性表达载体。为后续转基因研究这种差异是否会造成WRI1基因在提高含油量作用上的差异作准备。通过双酶切和连接反应,分别将WRI1基因和napin启动子基因插入到pBI121载体的GUSA基因和CaMV35启动子基因位置。酶切及PCR检测结果显示,napin启动子和WRI1已插入相应位置,完成2种载体种子特异性表达WRI1基因重组载体pBI121+napin+WRI1的构建,命名为pBI121NW2、pBI121NW4。     

高等植物ACC氧化酶基因启动子研究进展

启动子是控制植物基因表达的重要DNA序列结构,本文从高等植物ACO基因启动子克隆、顺式作用元件分析及GUS基因表达等方面着眼,综述该领域的研究进展。     

施氏鲟幼鱼摄食和生长的最适水温

实验用施氏鲟(AcipenserschrenckiiBrandt)幼鱼为黑龙江捕获的野生亲本经人工繁殖、培育所得,8月龄,体长(32 63±1 80)cm,实验周期35d。实验设计了4个水温处理组,实验水温分别为17℃、20℃、23℃和26℃,实验观察到,生活在不同水温环境中的幼鱼表现出不同的生长特性,养殖在23℃温度环境中的施氏鲟幼鱼的生长速度显著高于其他处理组幼鱼,水温对施氏鲟的特定生长率(SGR)、日增重(DWG)、食物转化率(FCR)和摄食率(FR)有着显著的影响。这些参数与水温(T)之间的相关关系可用二次回归曲线来描述,生长率与水温的回归方程式为:SGR=-12 933+1 418T+(-0 033)T2,摄食率与水温的关系式为:FR=-5 324+0 719T+(-0 016)T2。根据回归方程,求得施氏鲟幼鱼的最适生长水温为21 53℃,最大摄食率的水温为22 33℃。结论认为,施氏鲟在水温17～26℃都可摄食生长,但在20℃时最适宜;同时证实,施氏鲟最适生长温度要低于其最适摄食温度。     

用地高辛标记的PCR－ELISA技术快速检测转基因鱼

取含有小鼠重金属螯合蛋白（mMT)基因启动子及人生长激素（hGH)基因的鱼样品，以Qiagene核酸纯化技术对鱼组织DNA进行纯化。常规PCR检测显示，mMT启动基因和hGH基因的PCR产物分别为240bp和130bp，与设计相符。PCR产物的核酸测序结果证实其扩增产物具特异性。地高辛-PCR（Dig-PCR)标记反应显示，2个基因均产生1条Dig标记的特异产物带。Dig-PCR-ELISA检测敏感性试验显示，对mMT启动子和hGH基因的检测敏感性可达10^-3，与常规PCR结合琼脂糖胶电泳检测方法相比，检测敏感性可提高至100-1000倍。试验证明，针对mMT和hGH基因所建立的Dig-PCR-ELISA技术特异可靠。