靶向食蟹猴NTCP基因的CRISPR/Cas9系统gRNA筛选

本研究旨在通过CRISPR/Cas9体外酶切法及细胞水平上的PCR扩增测序筛选出靶向食蟹猴NTCP基因具有高敲除活性的gRNA。首先通过比对食蟹猴与人类NTCP氨基酸序列,选择差异位点,即第84-87位和第157-165位氨基酸作为基因靶点序列区;利用gRNA软件设计针对上述基因靶点序列的gRNA,每个靶点设计3~4条候选gRNA序列;然后利用gRNA体外检测试剂盒,筛选出靶向NTCP基因的体外敲除活性较高的两条gRNA序列：gRNA1.2和gRNA2.1。将gRNA1.2和gRNA2.1分别插入pLV hUbC-Cas9-T2A-GFP载体中,转染食蟹猴原代肝细胞。提取转染后细胞基因组DNA,通过PCR扩增NTCP基因并将其克隆到T载体中进行测序分析。结果表明,gRNA1.2和gRNA2.1均可使NTCP基因产生移码突变,但gRNA2.1比gRNA1.2具有更高的敲除活性。本研究为下一步编辑食蟹猴NTCP基因及研究其在乙型肝炎病毒（HBV）感染中的功能奠定了基础。     

恒河猴、食蟹猴毛发氨基酸和微量元素测定分析

目的:了解人工饲养环境条件下恒河猴、食蟹猴氨基酸和微量元素含量的基础值,为保障恒河猴、食蟹猴的饲养品质提供基础数据。方法:分别采集24只(雌雄各半)成年的恒河猴、食蟹猴被毛,测定其氨基酸及微量元素的含量,并进行比较分析。结果:毛发中丙氨酸(ALA)和铜的含量,食蟹猴组显著高于猕猴组(P0.05);异亮氨酸(ILE)的含量,恒河猴组极显著高于食蟹猴组(P0.01);亮氨酸(LEU)的含量,恒河猴组显著高于食蟹猴组(P0.05);其余氨基酸和微量元素的含量均无显著性差异(P0.05)。不同性别恒河猴、食蟹猴19种氨基酸和5种微量元素均无显著性差异(P0.05)。     

高脂高糖联合氧化剂tBHP诱导胰岛素抵抗2型糖尿病模型的建立与稳定性观察

为了建立高效稳定的胰岛素抵抗2型糖尿病(T2DM)模型并观察该模型的稳定性,试验以高脂高糖联合氧化剂叔丁基过氧化氢(t BHP)0.005 m L/g诱导建立胰岛素抵抗T2DM模型,测定SD大鼠不同时间点的体重、空腹血糖(FBG)、三酰甘油(TG)、总胆固醇(CHO)、胰岛素(FINS)、胰岛素敏感指数(ISI),将24 h FBG≥10.0 mmol/L及ISI≤正常动物平均值的大鼠定为T2DM模型大鼠;然后连续3 d给T2DM模型大鼠灌胃盐酸二甲双胍片75 mg/kg,研究该模型的有效性。结果表明:4周后,与对照组、双高组相比,t BHP组大鼠体重、FBG、TG、CHO、FINS、ISI均差异显著(P0.05);该T2DM模型在4周内非空腹血糖(NFBG)水平较高;盐酸二甲双胍片能降低该T2DM模型的NFBG。说明建立的胰岛素抵抗T2DM模型具有易行、稳定的特点。     

食蟹猴肺炎球菌病的诊断与防治

广西某野生动物保护站下属养猴场发生一起食蟹猴其症状呈急性经过、死亡率高的疾病，经临床观察、病理剖检及实验室检验，确诊为食蟹猴肺炎球菌病。１发病情况该养猴场于１９９８年４月２３日凌晨，从中越边境长途运输引进一批野外捕获的食蟹猴共１２４只，在到场６个小时...     

食蟹猴体温、呼吸率、血压值和脉搏数的测定

为提供食蟹猴一般生理指标,本实验测定124只食蟹猴的肛温、呼吸率、血压和脉搏,比较不同性别和不同年龄组之间测定参数的差异.结果表明,1～1.8岁青幼年组、3～6岁成年组食蟹猴的体温、呼吸率、血压和脉搏均无明显性别差异.青幼年组与成年组相比较,血压无明显差异,但肛温、呼吸率明显高于成年组食蟹猴,而脉搏则明显低于成年组(P<0.01) .结论:食蟹猴上述生理参数性别之间差异不明显,随着年龄的变化血压变化不大,但肛温和呼吸率随着年龄的增长而减少,脉搏数则随着年龄的增长而增多,呈现较为复杂的年龄相关性.     

2型糖尿病食蟹猴白细胞NF-κB和IκBα表达相关性分析

比较了食蟹猴2型糖尿病(T2DM)不同发病阶段与正常对照组的空腹血糖值(FPG)、甘油三酯(TG)、胆固醇(CHOL),低密度脂蛋白胆固醇(LDL-C),高密度脂蛋白胆固醇(HDL-C)在血液中含量,及荧光定量PCR方法测定的NF-κB和IκBα在外周血白细胞中表达量.结果显示,FPG、CHOL和LDL-C随疾病进行呈...     

微生态制剂对食蟹猴生长性能的影响

目的:研究微生态制剂不同添加水平对食蟹猴生长性能的影响.方法:分别采用0.5 g/kg、1.0 g/kg、2.0 g/kg的微生态制剂诱导的食蟹猴生长性能为模型,检测微生态制剂饲用后食蟹猴月增重、月采食量、料重比.结果:1.0 g/kg、2.0 g/kg的微生态制剂添加组与对照组相比,月增重均显著提高(P<0.05),料重比均显著降低(P<0.05).结论:食蟹猴基础饲粮中添加1.0 g/kg、2.0 g/kg的微生态制剂均能显著提高食蟹猴的生长性能.     

昆明地区食蟹猴种群繁殖规律与繁殖率的分析

试验旨在探究目前昆明地区群体养殖条件下食蟹猴的繁殖规律与繁殖率,统计昆明地区2018—2022年食蟹猴的妊娠、出生的高低峰期及妊娠率、成活率、繁殖率。结果显示,2018—2022年间,每年下半年7～10月、12月为昆明地区食蟹猴妊娠高峰期,而每年3～5月为妊娠低谷期。2018—2022年间,每年均有连续生产,2～5月为食蟹猴出生的高峰期,6～10月为出生低谷期。昆明地区食蟹猴平均妊娠率、成活率和繁殖率分别为80.40%、90.18%和72.44%,与其他地域数据相比偏高。研究表明,每年7～12月应对雌猴更加重视,每年2～3月、5月和12月应加强对新生猴的重视。数据结果可为昆明地区食蟹猴的繁殖规律、繁殖性能和生物学的研究提供参考,并为食蟹猴养殖业的发展提供帮助。     

蓝刺头及蓝刺头复方水提物对实验性Ⅱ型糖尿病大鼠血糖及血脂的影响

为研究蓝刺头及蓝刺头复方水提物对Ⅱ型糖尿病(T2DM)大鼠血糖、血脂的影响。本试验以SD大鼠饲喂高脂高糖饲料加小剂量链脲佐菌素(STZ)腹腔注射的方法建立T2DM大鼠模型。将大鼠随机分为正常对照组,模型组,蓝刺头水提物高、中、低浓度组,蓝刺头复方水提物高、中、低浓度组,对各组大鼠分别连续56d灌服生理盐水及相应浓度蓝刺头及蓝刺头复方水提物,并检测大鼠相关血糖血脂生化指标。结果显示,蓝刺头及蓝刺头复方水提物高、中剂量组对Ⅱ型糖尿病大鼠空腹血糖均呈明显升高趋势(P<0.05);蓝刺头及蓝刺头复方低剂量组对Ⅱ型糖尿病大鼠空腹血糖均无明显的改善;蓝刺头及蓝刺头复方水提物高、中、低剂量组均可明显降低TC、TG和LDL-C的含量(P<0.05),HDL-C无明显差异(P>0.05)。结果表明,蓝刺头及蓝刺头复方水提物对实验性Ⅱ型糖尿病大鼠具有降糖、降脂的作用。     

两种高脂高糖乳脂配方复制高脂血症大鼠模型的比较

比较两种高脂乳脂配方复制高脂血症模型大鼠的血脂、肝重及肝病理的变化,为高血脂症的预防、治疗以及研究选择合适的动物造模方法。比较两次试验中,正常对照1组和高脂高糖乳剂1组、正常对照2组和高脂高糖乳剂2组SD大鼠,10只/组,按10mL/kg分别给予相应的高脂高糖乳剂连续造模40d,正常对照组则给予等量纯净水,分别于造模前、造模10d、造模40d时测定大鼠体重、血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDLC)、低密度脂蛋白胆固醇(LDLC)含量及HDLC/TC比值,并测定肝脏重量以及肝脏病理组织学的变化。结果显示,与相应的正常对照组比较,第一次试验中的高脂高糖乳脂1配方造模40d,可使血脂含量升高,肝脏细胞脂肪变性,第二次试验中的高脂高糖乳脂2配方造模40d,只能使血脂含量升高,未见肝细胞脂肪变性。与高脂高糖乳脂1组相比,高脂高糖乳脂2组造模40d体重下降幅度较小(P0.05),TC、TG、LDLC含量升高幅度较少(P0.05),HDLC/TC比值降低幅度较少(P0.05),肝脏重量增大幅度较大(P0.05),肝脏系数增大幅度较少(P0.05),肝脏未产生肝细胞脂肪变性。结果表明,第一次试验中的高脂高糖乳剂1配方可以制作混合型高脂血症伴脂肪肝大鼠模型,第二次试验中的高脂高糖乳脂2配方可以制作混合型高脂血症大鼠模型。     

SD大鼠2型糖尿病模型的建立及相关指标的测定

通过链脲佐菌素(STZ)结合高糖高脂饮食诱导建立2型糖尿病大鼠模型。30只健康雄性SD大鼠随机分为模型组(20只)、空白组(10只)。模型组喂饲高糖高脂饲料4周后,用STZ 30mg/kg一次性左下腹腔注射。检测试验第1周、第4周,注射STZ后第1周、第4周、第8周、第12周空腹体重、血清葡萄糖(BG)、甘油三酯(TG)、胆固醇(TC)、胰岛素(INS)、葡萄糖依赖性促胰岛素释放肽(GIP)、胰岛素样生长因子1(IGF-1)、胰高血糖素样多肽-1(GLP-1)等指标,进行统计分析。处死大鼠,取胰腺进行病理切片检查。动物成模率为75%。试验第4周,模型组TG、TC值明显升高,与空白组比较,P<0.01。注射STZ后第1、4、8、12周,模型组BG、TG、TC值均明显升高,与空白组比较,P<0.01;ISI均明显降低,与空白组比较,P<0.01。注射STZ后第1、4、8周,模型组的GLP-1、IGF-1值明显降低,与空白组比较,P<0.01。注射STZ后第12周,模型组的GLP-1、GIP、IGF-1值明显降低,与空白组比较,P<0.01。试验结果表明,STZ一次性左下腹腔注射结合高糖高脂饮食可成功诱导2型糖尿病大鼠模型。GIP、IGF-1、GLP-1等细胞因子参与了2型糖尿病的病理发生。     

SD大鼠2型糖尿病动物模型的建立及胰腺组织SUR1 mRNA的表达

[目的]链脲佐菌素（streptozotocin,STZ）结合高糖高脂饮食诱导建立2型糖尿病大鼠模型,检测体重、血糖、胰岛素、胰岛素敏感指数（ISI）、胰腺组织SUR1 mRNA表达的变化。[方法]30只健康雄性SD大鼠随机分为模型组（20只）、空白组（10只）。模型组喂饲高糖高脂饲料4周后,用STZ 30mg/（kg·bw）一次性左下腹腔注射。检测注射STZ后第1周、第4周、第8周、第12周空腹体重、血糖、胰岛素、胰岛素敏感指数等指标,进行统计分析。剖杀大鼠,取胰腺,RT-PCR方法检测胰腺组织SUR1 mRNA的表达。[结果]动物成模率为75%。注射STZ后1、4、8、12周,模型组血糖值均明显升高,与空白组比较,P〈0.01;ISI均明显降低,与空白组比较,P〈0.01。模型组SUR1 mRNA表达显著低于对照组（P〈0.05）。[结论]STZ一次性左下腹腔注射结合高糖高脂饮食可成功诱导2型糖尿病大鼠模型。SUR1 mRNA的降低可能是糖尿病发病的分子机制之一。     

裂叶荨麻提取物对2型糖尿病小鼠模型糖脂代谢影响的研究

研究裂叶荨麻提取物对高脂高糖饮食联合链脲佐菌素(STZ)诱导2型糖尿病小鼠模型糖脂代谢的影响。将造模成功的糖尿病小鼠随机分为空白组,模型组,阳性组,荨麻提取物低、中、高剂量组(0.5、1、2 g/kg)。各组小鼠灌胃给药35 d,期间测定小鼠的体重、饮食量、饮水量、空腹血糖(FBG),末次给药结束后,测定血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)含量。连续灌胃治疗35 d后,小鼠的体重下降、多饮多食症状得到改善,荨麻提取物低、中、高剂量组FBG值分别下降了4.29、7.41、7.16 mmoL/L。与模型组相比,荨麻提取物各组小鼠TC值分别下降了1.17、0.76、1.77 mmoL/L,TG值分别下降了0.53、0.60、0.71 mmoL/L,LDL-C值分别下降了0.50、0.39、0.65 mmoL/L,HDL-C值分别升高了0.66、1.13、1.24 mmoL/L,其中高剂量组效果最为显著(P<0.01)。结果表明,裂叶荨麻提取物对2型糖尿病小鼠的高血糖及高血脂症状有一定的缓解作用。     

Effect of fat supplementation during transition period on plasma leptin and non‐esterified fatty acid concentrations in Holstein cows

The objective of this experiment was to investigate the effect of fat supplementation during the transition period on pre and postpartum body weight (BW), body condition score (BCS), non‐esterified fatty acids (NEFA), glucose and leptin concentrations in Holstein cows. Holstein cows (n = 15) received a low fat diet (LF; 1.61 Mcal net energy for lactation (NEL)/kg of dry matter [DM]), moderate fat diet (MF; 1.68 Mcal NEL/kg DM) or a high fat diet (HF; 1.74 Mcal NEL/kg DM) for 4 weeks prior to calving. All cows were fed similar lactation diets ad libitum (1.74 Mcal NEL/kg DM) for 30 days after calving. Increasing diet energy density during transition period had no effect on prepartum DMI, BCS, BW, glucose and NEFA concentrations (P > 0.05); but leptin concentrations and energy balance (EB) were affected by treatments (P < 0.05). Animals fed HF had less plasma leptin prepartum. After parturition, BW, milk production, milk fat, protein, urea nitrogen and plasma glucose concentrations were affected by prepartum diets (P < 0.05). Fat supplementation prepartum did not affect postpartum NEFA. In conclusion, prepartum fat supplementation decreased leptin concentration prepartum.     

Corticosterone administration and high-energy feed results in enhanced fat accumulation and insulin resistance in broiler chickens

1. Two experiments were conducted to investigate the effects of exogenous corticosterone administration (30 mg/kg diet) and dietary energy level on feed or energy intake and fat deposition in broiler chickens of 1 and 4 weeks of age. 2. Corticosterone treatment significantly suppressed body weight (BW) gain and reduced feed and caloric efficiencies. The retarded growth may conceal the stimulatory effect of corticosterone on feed consumption or metabolisable energy (ME) intake. A high-energy diet may increase energy intake and partially alleviate the suppressing effect of corticosterone on growth of broilers. 3. Corticosterone administration promoted the conservation of energy stores as fat at both abdominal and subcutaneous sites and this process occurred regardless of dietary energy level in ad libitum feeding status. A high-energy diet increased fat accumulation and showed no significant interaction with corticosterone treatment. 4. The suppressed development of breast and thigh muscles by corticosterone treatment was observed only in 1-week-old chickens fed on the low-energy diet. In contrast, the yield of breast muscle but not thigh muscle was significantly decreased by corticosterone in 4-week-old chickens, suggesting that the tissue specificity to corticosterone challenge is age dependent. 5. Plasma concentrations of glucose, insulin, triglyceride, non-esterified fatty acids (NEFA) and very low density lipoprotein were increased by corticosterone treatment regardless of diet treatment. A high-energy diet increased plasma levels of NEFA and resulted in hyperinsulinism in 4-week-old chickens but not in 1-week-old chickens. 6. Lipoprotein lipase (LPL) activities in adipose tissues may have been up-regulated by corticosterone treatment and showed tissue specificity. The increased LPL activities at ad libitum feeding status were not necessarily linked with the increased fat accumulation in corticosterone challenged chickens. 7. Corticosterone resulted in augmented energy consumption and altered energy redistribution toward lipid deposition. The induced insulin resistance and enhanced hepatic de novo lipogenesis by corticosterone are likely to be responsible for the increased fat deposition.     

Effect of bypass fat supplementation on productive performance and blood biochemical profile in lactating Murrah (Bubalus bubalis) buffaloes

The study investigated the effect of dietary supplementation of bypass fat on productive performance and blood biochemical profile of lactating Murrah buffaloes (Bubalus bubalis). Fifteen multiparous buffaloes (2-4 lactation) of early to mid lactation were divided in three homogenous groups T(1) (control), T(2), and T(3) of five each. The animals in T(1) were fed with a basal diet consisting of a concentrate mixture, green sorghum, and wheat straw as per requirements, while the animals in group T(2) and T(3) were fed with same ration supplemented with 0.7 % (100?g/day) and 1.4 % (200?g/day) bypass fat (on dry matter intake (DMI) basis), respectively. The feed intake, milk yield, and milk composition were not influenced by supplemental bypass fat. However, fat-corrected milk (6.5 %) yield was higher (P?相似文献   

高脂饲粮和高碳水化合物饲粮对大鼠脂肪代谢的影响

本试验旨在以相同代谢能水平为基础,研究高脂饲粮和高碳水化合物饲粮对大鼠脂肪代谢的影响。选取48只8周龄雄性SD大鼠,随机分为3组,每组8个重复,每个重复2只。3个组分别饲喂高脂饲粮(HF组)、高碳水化合物饲粮(HC组)和对照饲粮(CON组),试验期9周。结果表明:1)与CON组相比,HF组、HC组大鼠的初重、末重、日增重均无显著差异(P0.05),日采食量极显著降低(P0.01),饲料转化效率极显著升高(P0.01)。2)各组大鼠的Lee’s指数、肾脏指数、胃指数、脾脏指数、肝脏指数无显著差异(P0.05)。3)与CON组和HF组相比,HC组大鼠血清甘油三酯、总胆固醇、高密度脂蛋白胆固醇含量显著或极显著升高(P0.05或P0.01);与HC组和CON组相比,HF组大鼠血清葡萄糖含量极显著增加(P0.01),血清尿素氮含量极显著降低(P0.01)。与CON组相比,HF组和HC组大鼠肝脏甘油三酯含量极显著升高(P0.01),HF组大鼠肝脏总胆固醇含量极显著升高(P0.01)。4)与CON组相比,HF组和HC组大鼠肝脏磷酸烯醇式丙酮酸激酶(PEPCK)mRNA表达量极显著增加(P0.01);与CON组和HC组相比,HF组大鼠肝脏胆固醇调节元件结合蛋白1(SREBP1)mRNA表达量极显著增加(P0.01)。综上所述,相同代谢能水平下,HF组和HC组大鼠体重无显著增加。HC组大鼠血脂升高;HF组大鼠血清葡萄糖、肝脏总胆固醇含量升高,血清尿素氮含量下降。HF组和HC组大鼠肝脏甘油三酯含量升高,且通过肝脏PEPCK和SREBP1基因表达调控途径影响脂肪代谢     

饲粮添加万寿菊提取物对肉鸡血清生化指标、抗氧化能力和免疫性能的影响

本试验旨在研究饲粮添加万寿菊提取物对肉鸡血清生化指标、抗氧化能力和免疫性能的影响。选取1日龄爱拔益加(AA)肉鸡192只,随机分为3组,每组8个重复,每个重复8只鸡。对照组饲喂基础饲粮,试验组在基础饲粮中分别添加0.15%和0.60%的万寿菊提取物,即有效成分叶黄素含量分别为30和120 mg/kg,试验期42 d。结果表明:与对照组相比,1)饲粮添加0.60%万寿菊提取物使肉鸡血清甘油三酯和低密度脂蛋白胆固醇水平显著降低(P0.05),高密度脂蛋白胆固醇水平显著升高(P0.05);2)0.15%和0.60%万寿菊提取物添加组肉鸡21和42日龄血清总抗氧化能力和超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶活性显著提高(P0.05);3)0.60%万寿菊提取物添加组肉鸡21日龄胸腺、脾脏和法氏囊指数显著升高(P0.05),饲粮添加万寿菊提取物使肉鸡血清免疫球蛋白M、免疫球蛋白G和白细胞介素2水平显著升高(P0.05),万寿菊提取物未对肉鸡42日龄免疫器官指数造成显著影响(P0.05)。综上,肉鸡饲粮中添加万寿菊提取物可以提高肉鸡机体的抗氧化能力和免疫性能,且对机体脂代谢有调节作用。     

Effects of additional starch or fat in late-gestating high nonstarch polysaccharide diets on litter performance and glucose tolerance in sows

The effects of feeding additional starch or fat from d 85 of gestation until parturition on litter performance and on glucose tolerance in sows that were fed a diet with a high level of fermentable nonstarch polysaccharides (NSP) were studied. The day after breeding, 141 multiparous sows were assigned to the experiment. At d 85 of gestation, sows were assigned to the treatments. Sows were fed 3.4 kg/d (as-fed basis) of a high-NSP diet or the same quantity of the high-NSP diet and an additional 360 g of starch (from wheat starch) daily, or the same quantity of the high-NSP diet and an additional 164 g of fat (from soybean oil) daily. During lactation, all sows were given free access to the same lactation diet. Approximately 1 wk before the expected time of parturition, an oral glucose tolerance test was performed in 38 randomly chosen sows by feeding pelleted glucose (3 g/kg BW0.75). Blood samples for glucose analyses were taken at -10, 10, 20, 30, 40, 50, 60, 70, 80, 90, 105, and 120 min after glucose was fed. The supply of additional dietary starch or fat did not increase piglet birth weight or total litter weight at birth. Sows that were fed the high-NSP diet had more (P = 0.097) live-born piglets and fewer (P = 0.084) stillborn piglets than did sows that were fed additional fat, whereas sows that were fed additional starch were intermediate for these variables. Piglet mortality after birth was not affected by dietary treatment. Body weight and backfat gains in the last month of gestation were higher for sows fed additional starch or fat than for sows fed the high-NSP diet (P < 0.001 and P = 0.017, respectively). Feed intake in lactation was greatest by sows fed the high-NSP diet, least by sows fed additional starch at the end of gestation, and intermediate by sows fed additional fat (P = 0.099). The differences in lactation feed intake did not result in differences in BW and backfat losses during lactation. Sows that were fed additional fat had the greatest glucose area under the curve (P = 0.044), indicating that these sows were less tolerant to glucose. In conclusion, feeding additional energy (starch or fat) in late-gestating sows that are fed a high-NSP diet did not increase litter weight at birth or piglet survival, but did increase maternal gain. Feeding sows additional energy from fat might induce glucose intolerance, whereas feeding sows additional energy from starch did not induce glucose intolerance.     

日粮添加镁、硒、铬对热应激肉仔鸡血清生化和血液流变学的影响

试验旨在研究热应激情况下镁、硒、铬联合添加对肉仔鸡血清生化指标和血液流变学的影响。选用240只1日龄AA肉公鸡,随机分为4个处理,每个处理6个重复。T1为常温对照组,T2、T3、T4为热应激组。T1、T2饲喂基础日粮,T3、T4在基础日粮中分别添加镁、硒、铬2000、1.5、1.5mg/kg和4000、3.0、3.0mg/kg。结果表明:T3、T4组热应激鸡只死亡率显著降低,血清TP、ALB、GLOB、HDL水平显著升高(P0.05),GLU、BUN、TC含量显著降低(P0.05)。同时,T3、T4组的GPT、CK酶活力及低切变率下的全血黏度均显著降低(P0.05)。试验表明,联合添加镁、硒、铬能有效改善热应激肉鸡的血液生化和血液流变学指标,减轻热应激对机体的损伤,从而降低鸡只死亡率。