透皮促渗剂对乳康涂膜剂抗炎消肿作用的影响

乳康涂膜剂是由蒲公英、金银花、连翘、丝瓜络、通草组成的复方中药涂膜剂.它具有清热解毒、理气活血、消肿散结、通经下乳等作用,主要用于治疗母畜乳房炎.为增强乳康涂膜剂的抗炎消肿作用,特设了不同透皮促进剂组合及配合增效剂和化学抗炎剂,进行抗炎消肿试验[3],报告如下.     

自制中草药复方透皮软膏对奶牛乳房炎治疗效果观察

为了研究中草药复方透皮软膏对奶牛乳房炎的治疗效果,试验选用超级头孢金品和速诺对奶牛最急性乳房炎、急性乳房炎、亚急性乳房炎和慢性乳房炎进行临床治疗对比试验。结果表明:自制的中草药复方透皮软膏对慢性乳房炎的疗效明显高于对照组,对亚急性乳房炎治疗效果显著。     

乳炎宁搽剂的安全性试验

乳炎宁搽剂是由土贝母、白芷、松香等中药与蓖麻油经特殊工艺制成的一种透皮吸收中药复方制剂,具有抗炎、祛瘀、消肿和止痛作用.临床用于治疗奶牛产前、产后乳房水肿和奶牛乳房炎.与传统的治疗乳房炎的方法相比,本制剂疗效确切、给药方便、无残留、无毒副作用,在临床试验中取得了明显的效果.为了探讨乳炎宁搽剂的安全性,进行了本试验.     

基于网络药理学与分子对接探究中兽药复方乳康颗粒治疗奶牛乳房炎的作用机制

【目的】探究中兽药复方乳康颗粒(RKG)治疗奶牛乳房炎的作用机制。【方法】通过中药系统药理学分析平台(TCMSP)、中医药百科全书(ETCM)平台获取中兽药复方乳康颗粒中黄芪、王不留行、益母草的有效成分及靶点,利用Cytoscape v 3.7.2构建中兽药复方乳康颗粒中药-成分-靶点网络图,从NCBI、OMIM、GeneCards、MalaCards平台获取奶牛乳房炎靶点,使用Cytoscape v 3.7.2中的Bisogenet插件对中兽药复方乳康颗粒成分靶点及奶牛乳房炎的靶点进行综合分析,构建蛋白互作(PPI)网络图,并使用DAVID进行GO功能和KEGG通路富集分析,探讨其治疗奶牛乳房炎的作用机制。采用分子对接对网络药理学预测结果进行初步验证。【结果】通过网络药理学筛选出槲皮素、山奈酚、异鼠李素等23个中兽药复方乳康颗粒相关的活性成分,文献检索得到奶牛乳房炎相关靶点有肿瘤坏死因子(TNF)、Toll样受体4(TLR4)、白细胞介素17A(IL17A)等49个,通过Bisogenet插件对其进行PPI分析后得到中兽药复方乳康颗粒作用于奶牛乳房炎的关键靶点77个,涉及到靶点肿瘤蛋...     

不同透皮促进剂对"乳炎克"促渗作用的研究

为研究复合型透皮促进剂对自制“乳炎克”的体外渗透作用，我们以奶牛乳房皮肤为试验屏障。小檗碱为检测指标，观察了氮酮、油酸、丙二醇、薄荷4种透皮吸收促进荆的促渗效果。     

中药透皮剂对隐性乳房炎奶牛泌乳量、白细胞及乳腺局部免疫的影响

为开发安全、有效、稳定的新型中兽药制剂,有利于奶牛隐性乳房炎的防治,试验研究了中药透皮剂对隐性乳房炎奶牛泌乳量、白细胞、白蛋白（ALB）、丙氨酸氨基转移酶（ALT）、乳酸脱氢酶（LDH）的影响。方法：记录用药前后5d日泌乳量,分别采用试剂盒和CA-500血液分析仪测定用药前后乳汁中ALB、ALT、LDH和血液中白细胞。结果：患隐性乳房炎奶牛泌乳量显著降低（P〈0．05）,血液中WBC、MON、GRA、LYM％显著高于健康牛（P〈0．05）,而乳汁中的ALB、ALT和LDH含量极显著高于健康奶牛（P〈0．01）。通过5d中药透皮剂治疗后,奶牛产奶量大幅度增加,WBC、MON、GRA、LYM％、ALB、ALT和LDH与健康牛比差异不显著（P〉0．05）。结论：中药透皮剂对奶牛隐性乳房炎具有良好的治疗效果。     

复方特比萘芬纳米乳的皮肤安全性及经皮渗透性试验

考察复方特比萘芬纳米乳的透皮特性及皮肤毒性,为其临床应用提供试验依据。应用皮肤刺激性试验、皮肤急性毒性试验及皮肤过敏性试验考察复方特比萘芬纳米乳的皮肤安全性;采用改良的Franz立式扩散池,对复方特比萘芬纳米乳进行体外经皮透过性评价。结果显示,复方特比萘芬纳米乳对家兔无皮肤刺激性和皮肤急性毒性,对豚鼠皮肤无致敏性。复方特比萘芬纳米乳中特比萘芬与其混悬液的透皮速率(J)分别为0.840 3μg/cm.h和0.266 8μg/cm.h。表明复方特比萘芬纳米乳是一种安全性较好的外用制剂,且纳米乳增强了特比萘芬的经皮透过性。     

中草药复方搽剂防治奶牛乳房炎的效果观察

试验用中草药复方透皮吸收搽剂防治奶牛乳房炎,共治疗临床型奶牛乳房炎18例.治愈16例,与抗生素联合用药治愈2例,治愈率达88.9%,有效率为100%:通过体细胞(SCC)的检测判断其对隐性乳房炎的影响.结果表明,试验所用中草药搽剂对降低牛乳中体细胞数,防治奶牛隐性乳房炎有着良好的效果.     

复方中药软膏剂的透皮效果研究

为评价中药复方软膏剂透皮效果,筛选最佳组方,以中药软膏剂中绿原酸成分为标记物,应用高效液相色谱法(HPLC)测定透过液中绿原酸的含量,评价不同的中药剂型和不同的透皮促渗剂对中药软膏剂透皮效果的影响。结果表明,中药超微粉直接入药并以含45 g/L冰片+45 mL/L氮酮作为促渗剂效果最佳。     

金黄超微透皮软膏对奶牛乳房炎的治疗效果观察

为了探讨金黄超微透皮软膏对奶牛乳房炎的治疗效果,将30头隐性乳房炎患牛和25头临床型乳房炎患牛分别各自随机分为2组。其中,隐性乳房炎患牛中的15头48个乳区为试验组,另15头42个乳区为对照组;临床型乳房炎患牛中的15头25个乳区为试验组,另10头15个乳区为对照组。试验组奶牛每天涂抹受试金黄超微透皮软膏2次,连用3~5d;对照组不用药。每天采集隐性乳房炎患牛奶样,测定其体细胞数变化情况;每天分别从外观、手感、乳汁性状和CMT4项指标评判其临床型乳房炎的治疗情况。结果显示,隐性乳房炎患牛治疗3d后,有效率为100%,治愈率为89.58%,而不用药的对照组无1例痊愈;临床型乳房炎患牛连续用药5d,停药1周后观测,试验组80%的乳区4项指标全部达痊愈水平,而对照组均未达痊愈水平。试验结果表明,金黄超微透皮软膏对奶牛隐性和临床型乳房炎均有良好的治疗效果。     

Low expression of the antibacterial factor L‐amino acid oxidase in bovine mammary gland

In the mouse, L‐amino acid oxidase (LAO) produces hydrogen peroxide by utilizing free amino acids and is a proven antibacterial factor in mammary glands. Mastitis, a bacterial infection of the mammary gland, is the most frequent disease in dairy cattle. Here, we investigate whether LAO is expressed in the mammary gland of dairy cattle and is antibacterial. In dairy cattle, the expression level of LAO mRNA in the mammary gland was considerably lower than that in mice, and LAO activity was not observed in cattle milk that produced hydrogen peroxide. The expression of LAO mRNA was also low in Japanese Black cattle, the same as in Holstein cattle. A higher LAO mRNA expression was observed in the mastitis glands than in the lactating glands. Furthermore, spleen and lymph nodes expressed high levels of LAO mRNA in dairy cattle. We conclude that mammary glands in dairy cattle have lower ability to express the LAO gene compared to that in mice, which may result in a high incidence of mastitis.     

金黄色葡萄球菌表面蛋白A对奶牛乳腺上皮细胞的黏附作用

本试验拟探究金黄色葡萄球菌（金葡菌）表面蛋白A（Staphylococcus aureus surface protein A，SasA）编码基因在奶牛乳源金葡菌中是否具有普遍性及同源性，并结合蛋白结构组成研究SasA对奶牛乳腺上皮细胞的黏附作用。试验前期从中国北方5个荷斯坦牛场无菌条件下分离纯化了73株奶牛乳源金黄色葡萄球菌菌株，PCR扩增鉴定SasA基因并进行序列保守性分析。利用原核表达系统表达SasA蛋白的丝氨酸富集片段1（serine-rich repeat region 1， SRR1）及非重复区域（non-repeat region， NRR）并进行蛋白纯化。利用流式细胞仪检测NRR，牛血清白蛋白（bovine serum albumin， BSA）及SRR1对奶牛乳腺上皮细胞（Mac-T）黏附性差异。为进一步在NRR中定位发挥主要黏附作用的片段，试验采用了长度不同的NRR片段与完整NRR片段做竞争性黏附处理。PCR扩增产物鉴定及序列同源性分析结果显示，86.3%的牛源金葡菌菌株含SasA基因且序列一致性超过95%。相较于SRR1和BSA，NRR对细胞具有更强的黏附性。黏附抑制试验结果表明，NRR1-2片段（230—540 aa）对NRR抑制作用最明显。综上表明，SasA基因在奶牛乳源金葡菌中具有普遍性，该基因序列具有高度的保守性。在SasA蛋白中，对奶牛乳腺上皮细胞起主要黏附作用的为NRR1-2片段，大致定位在其结构域的230—540位氨基酸。本研究结果表明，与该区域结合的受体中可能存在SasA作为黏附素与奶牛乳腺上皮细胞互作的位点。     

干扰PTEN基因对奶山羊乳腺上皮细胞脂质合成相关基因的转录及脂肪酸组成的影响

旨在通过RNA干扰技术揭示蛋白酪氨酸磷酸酶（phosphatase and tensin homolog deleted on chromosome 10，PTEN）基因在奶山羊乳腺上皮细胞中对乳脂合成及脂肪酸组成的影响。利用RT-PCR方法扩增到西农萨能奶山羊乳腺组织中PTEN基因（GenBank登录号：MK158074.1）的CDS区，进行序列分析和不同泌乳时期差异转录分析。合成靶向该基因的siRNA，由RT-qPCR结果筛选出有效siRNA，将其转染至奶山羊的乳腺上皮细胞，进一步检测干扰该基因后对脂质合成相关基因转录水平及脂肪酸组成的影响。结果表明：本研究首次克隆到奶山羊（Capra hircus）PTEN基因的CDS区，全长为1 212 bp；经序列比对发现，山羊的PTEN基因核苷酸序列同牛（Bos taurus）、猪（Sus scrofa）和人（Homo sapiens）的相似度分别为99%、98%和97%，编码氨基酸序列相似性均在99%以上；蛋白质结构预测发现：其蛋白不存在跨膜结构域，亚细胞定位于细胞质中，N端具有保守的磷酸酶功能区；该基因在奶山羊泌乳盛期乳腺组织中的转录量较干奶期下降51.5%；合成的siRNA转染至乳腺上皮细胞后，成功筛选出理想的siRNA，干扰效率达到94%（P<0.01）；与对照组相比，干扰PTEN基因后显著上调SREBP1、FASN、ACACA、SCD1基因转录量（P<0.01或P<0.05），显著下调LPL、FABP3、ACOX1、CPT1B、GPAM、DGAT2和HSL基因转录量（P<0.01或P<0.05）。脂肪酸检测分析发现：干扰该基因能够显著上调C16：1的去饱和指数（P<0.05），但对C18：1的去饱和指数没有显著影响。综上所述，PTEN基因能够调控乳腺上皮细胞中脂质合成相关基因的转录及脂肪酸组成，在奶山羊乳脂代谢中发挥重要作用。     

生物抗氧化剂对原料奶体细胞数的影响

原料奶中体细胞是间接反应奶牛乳房是否健康的一个有力数据。同时过高的体细胞也将直接影响到原料奶的质量和价格。因此,本试验选择年龄、胎次、产奶量、泌乳天数相近的中国荷斯坦奶牛80头,探讨日粮添加34g生物抗氧化剂对原料奶中体细胞的影响。结果表明,生物抗氧化剂对奶牛产奶量及乳成分无显著影响。但与对照组相比,试验组原料奶中体细胞数呈显著低于对照组（P〈0.05）。因此,本研究表明,生物抗氧化剂能够有效降低原料奶中体细胞数,提高原料奶质量。     

Proteomic analysis to unravel the effect of heat stress on gene expression and milk synthesis in bovine mammary epithelial cells

Heat stress can play a negative effect on milk yield and composition of dairy cattle, leading to immeasurable economic loss. The basic components of the mammary gland are the alveoli; these alveolar mammary epithelial cells reflect the milk producing ability of dairy cows. In this study, we exposed bovine mammary epithelial cells to heat stress and compared them to a control group using isobaric tags for relative and absolute quantitation combined with liquid chromatography coupled with tandem mass spectrometry. Compared with a control group, 104 differentially elevated proteins (>1.3‐fold) and 167 decreased proteins (<0.77‐fold) were identified in the heat treatment group. Gene Ontology analysis identified a majority of the differentially expressed proteins are associated in cell‐substrate junction assembly, catabolic processes and metabolic processes. Some of these significantly regulated proteins were related to the synthesis and secretion of milk, such as milk protein and fat. This finding was further supported by the results obtained from the reduced β‐casein expression through the system of plasminogen activator – plasminogen – plasmin and decreased fatty acid synthase could partly explain why milk fat synthesis ability of dairy cows decreased under heat stress. Our results highlight the effects of heat stress on synthesis of milk protein and fat, thus providing additional clues for further studies of heat stress on dairy milk production.     

研究不同氨基酸模式对奶牛乳腺上皮细胞生长的影响

本试验以泌乳奶牛乳腺组织为原料,采用组织块培养技术,研究培养基内不同氨基酸模式对奶牛乳腺上皮细胞增殖的影响。结果表明,随着培养天数的增加,细胞数量逐渐增加,第10天各组细胞数量分别达到最高,乳腺上皮细胞生长曲线呈"S"形。不同氨基酸模式对乳腺细胞生长有显著影响,与氨基酸不平衡组相比,氨基酸平衡组、理想组更能促进细胞生长,各组间差异显著(P0.05)。     

Pneumothorax in dairy cattle: 30 cases (1990-2003)

OBJECTIVE: To evaluate the features, underlying causes, results of diagnostic testing, and treatment of pneumothorax in dairy cattle. DESIGN: Retrospective study. ANIMALS: 30 dairy cattle. PROCEDURE: Medical records of all cattle with a diagnosis of pneumonia were reviewed. For cattle with pneumothorax, information was obtained pertaining to signalment, anamnesis, diagnosis, treatment, and outcome. Survival data were compared between cattle with pneumonia with or without pneumothorax. RESULTS: Pneumothorax was associated with bronchopneumonia in 18 cattle, interstitial pneumonia in 7 cattle, pharyngeal or laryngeal trauma in 3 cattle, and neonatal respiratory distress in 2 calves. Bovine respiratory syncytial virus was the most commonly detected infectious agent. Eighteen of 30 (60%) cattle survived; 8 were euthanatized and 4 died. Survival rate was 81% for cattle with pneumonia without pneumothorax during the same time period. Pneumothorax was a significant risk factor for failure to survive to discharge from the hospital for cattle with underlying chronic bronchopneumonia. CONCLUSIONS AND CLINICAL RELEVANCE: Pneumothorax in dairy cattle appears to occur most commonly in association with chronic bronchopneumonia. Cattle of both sexes and all ages can be affected.     

Serum bile acid concentrations in clinically normal cattle: comparison by type, age, and stage of lactation.

Serum total bile acid concentrations were determined for various types and ages of cattle. There was extreme variability among all the cattle, but the variance was twice as large (0.50 vs 0.22 in logarithmic scale) for beef cattle than for dairy cattle. There was no significant difference in serum total bile acid concentrations between beef cattle and dairy cattle in midlactation. Values for calves < 6 weeks old and for 6-month-old heifers were significantly (P = < 0.05) lower than values for lactating dairy cows. The 5th to 95th percentile range of values (mumol/L) for beef cattle was 9 to 126; for lactating dairy cattle, 15 to 88; and for 6-month-old dairy heifers, 11 to 64.     

Association between Coxiella burnetii shedding in milk and subclinical mastitis in dairy cattle

The objective of this research was to explore the potential association between Coxiella burnetii shedding in milk and chronic subclinical mastitis in dairy cattle. In two separate studies, we identified an association between PCR-based detection of C. burnetii in milk and chronic subclinical mastitis in lactating dairy cows. These studies were conducted in a commercial dairy herd where there was ongoing intensive monitoring of subclinical mastitis by aerobic bacteriology, but no prior knowledge or management of C. burnetii infections. In a case-control study, quarter level C. burnetii status determined by real-time quantitative PCR (RT-qPCR) was strongly associated with chronic subclinical mastitis as measured by milk somatic cell counts. In a subsequent cross sectional study, 147 (45%) of 325 lactating cows were positive for C. burnetii by RT-qPCR of composite milk samples. In a generalized linear model, accounting for the effect of covariates including aerobic intramammary infection status, C. burnetii PCR status was a significant predictor of linear somatic cell count score. In agreement with a small number of previous reports, this research provides evidence that there may be mammary gland specific manifestations of C. burnetii infections in dairy cattle.     

甘肃、宁夏地区奶牛衣原体血清流行病学调查及风险因素分析

本研究旨在查明甘肃与宁夏地区奶牛衣原体的感染情况并分析影响其感染的风险因素。本试验采用间接血凝试验(IHA)方法检测了甘肃榆中(751份)、宁夏青铜峡(450份)和宁夏吴忠(456份)3个地方总计1 657份奶牛血清样品,并应用流行病学调查及统计学方法对影响奶牛衣原体感染的因素进行了分析。流行病学调查结果显示,奶牛衣原体抗体总阳性率为29.33%;应用logistic回归分析评估奶牛衣原体感染的风险因素,结果显示年龄和胎次不是显著风险因素(P>0.05),而地区因素是影响奶牛衣原体感染的风险因素(P<0.05)。奶牛衣原体抗体滴度最高达1:1 024。结果表明,甘肃和宁夏地区奶牛衣原体普遍流行。因此,应当提高对调查地区奶牛衣原体感染的重视,采取适当的综合控制方法和有效的管理措施以防控奶牛衣原体病,以保证奶牛养殖业的经济效益。