紫锥菊通过调控TLR4-NF-κB通路增强免疫抑制鸡的免疫功能

紫锥菊是一种重要的免疫增强剂,常作为替抗药物用于饲料添加剂,但其作用机制有待进一步研究。本试验旨在研究紫锥菊对免疫抑制鸡免疫功能的影响及其机制。将120羽7日龄的雏鸡随机均分为：对照组、环磷酰胺组、紫锥菊组、紫锥菊+环磷酰胺组(联合组)。环磷酰胺组和联合组雏鸡连续3 d胸肌注射生理盐水配制的环磷酰胺溶液(80 mg·kg^-1),同时对照组和紫锥菊组连续3 d注射等量生理盐水,除紫锥菊组和联合组每天饲喂含紫锥菊全草粉末(含量为1%)的基础日粮外,其余各组雏鸡饲喂基础日粮,每天称重并记录,连续饲喂21 d。试验结束时,分析体重及脾脏指数的变化,检测炎性因子(NF-κB、IκB、IL-2、IL-4、IL-6、TNF-α、IFN-γ)和TLR家族(TLR2、TLR4、TLR7、MyD88)基因mRNA及蛋白的表达含量。结果显示,紫锥菊处理提高了雏鸡的生长性能,环磷酰胺抑制了雏鸡的生长并显著降低了脾脏系数(P<0.05),且环磷酰胺组脾组织病理变化明显,而在联合组中上述病理变化得到缓解。与对照组相比,环磷酰胺组的NF-κB、IL-2、IL-4、IL-6、IFN-γ、TLR2、TLR4、TLR7、MyD88的mRNA水平均显著下调(P<0.05、P<0.01或P<0.001),而联合组相比于环磷酰胺组,除TLR7外,上述基因mRNA表达量均显著上调(P<0.05、P<0.01或P<0.001)。此外,紫锥菊的添加显著提高了炎性因子和TLR家族相关蛋白的表达水平。结果表明：紫锥菊可通过调控TLR4/NF-κB通路增强免疫抑制鸡的免疫功能。     

蛹虫草多糖对注射环磷酰胺小鼠抗氧化功能的影响

试验旨在阐明蛹虫草多糖（CMP）对小鼠抗氧化活性的影响。选取90只雄性BALB/c小鼠,体重18~20 g,随机分为6组,3个CMP组（低、中、高CMP组）,模型对照组（CY组）、阳性对照组（PC组）及空白对照组（BC组）各1个。CMP组和CY组分别腹腔注射80 mg/kg环磷酰胺,连续3 d,然后3个CMP组分别灌胃给予17.5、35.0、70.0 mg/kg体重的CMP,BC组和CY组给予生理盐水,PC组灌胃给予70 mg/kg体重的CMP,持续18 d。于最后一次给药24 h后,采集小鼠内脏,测定心脏、肝脏及肾脏匀浆液中T-AOC、MAD、CAT、SOD和GSH-Px水平。结果显示,与CY组相比,3个CMP处理组小鼠内脏匀浆液中的SOD和T-AOC活性均极显著增加（P<0.01）,MDA水平极显著降低（P<0.01）;低剂量CMP组心脏CAT无显著变化（P>0.05）,中、高剂量CMP组心脏CAT水平分别显著（P<0.05）、极显著（P<0.01）提高;除低剂量CMP组肝脏GSH-Px活性显著增加（P<0.05）外,其余CMP组内脏GSH-Px活性均极显著增加（P<0.01）。本试验中CMP可以有效提高小鼠心脏、肝脏及肾脏的抗氧化活性,这为抗氧化新药的研发提供了借鉴。     

太子参多糖对环磷酰胺所致肠道黏膜损伤小鼠SIgA、IL-2、IL-6含量的影响

试验将36只18~22 g雄性昆明小鼠随机分为空白对照组、太子参多糖对照组(400 mg/kg体重)、环磷酰胺(CY)模型组、太子参多糖低、中、高剂量组(100、200、400 mg/kg体重),上述各组分别灌喂蒸馏水和多糖,连续灌胃19 d,第20天,除空白对照组和太子参多糖对照组小鼠腹腔注射生理盐水外,其余4组均腹腔注射CY(100 mg/kg体重),24 h处死小鼠,取十二指肠和回肠,放免法测定分泌型免疫球蛋白A(SIgA)、白细胞介素-2(IL-2)、白细胞介素-6(IL-6)的含量,研究太子参多糖对CY所致肠道黏膜损伤小鼠中SIgA、IL-2、IL-6分泌的影响。结果显示,与CY模型组相比,太子参多糖高、中剂量组十二指肠SIgA含量显著升高(P<0.05),太子参多糖高剂量组回肠SIgA含量极显著升高(P<0.01);太子参多糖高剂量组中十二指肠和回肠IL-2含量显著升高(P<0.05),十二指肠IL-6含量极显著升高(P<0.01),回肠IL-6含量显著升高(P<0.05)。上述结果表明太子参多糖在一定程度上能颉颃CY所致的肠道黏膜免疫损伤。     

共轭亚油酸对环磷酰胺免疫抑制肉仔鸡巨噬细胞溶菌酶含量的影响

本研究采用健康肉仔鸡和环磷酰胺免疫抑制肉仔鸡模型,通过体外细胞培养试验来研究2种共轭亚油酸异构体（c9,t11-CLA和t10,c12-CLA）对肉鸡巨噬细胞溶菌酶分泌水平的影响。选取24只1日龄爱拔益加(Arbor Acre )肉仔鸡随机分为免疫抑制处理组和健康肉仔鸡组,分别于14、15和16日龄进行环磷酰胺（cyclophosphamide,CY;80 mg/kg体重）腿肌注射和等体积生理盐水腿肌注射。巨噬细胞体外培养试验采用含不同浓度（0、25、50和100 μmol/L）c9,t11-CLA、t10,c12-CLA或1︰1的混合物（CLA混合物）。结果表明,t10,c12-CLA可减缓CY对巨噬细胞溶菌酶合成和分泌的抑制作用。添加高剂量的t10,c12-CLA效果好于低剂量添加。c9,t11-CLA对巨噬细胞溶菌酶的分泌无明显影响。     

Comparison of doxorubicin–cyclophosphamide with doxorubicin–dacarbazine for the adjuvant treatment of canine hemangiosarcoma

Canine hemangiosarcoma (HSA) is a neoplasm of vascular endothelial origin that has an aggressive biological behaviour, with less than 10% of dogs alive at 12‐months postdiagnosis. Treatment of choice consists of surgery followed by adjuvant doxorubicin‐based chemotherapy. We prospectively compared adjuvant doxorubicin and dacarbazine (ADTIC) to a traditional doxorubicin and cyclophosphamide (AC) treatment, aiming at determining safety and assessing whether this regimen prolongs survival and time to metastasis (TTM). Twenty‐seven dogs were enrolled; following staging work‐up, 18 were treated with AC and 9 with ADTIC. Median TTM and survival time were longer for dogs treated with ADTIC compared with those receiving AC (>550 versus 112 days, P = 0.021 and >550 versus 142 days, P = 0.011, respectively). Both protocols were well tolerated, without need for dose reduction or increased interval between treatments. A protocol consisting of combined doxorubicin and dacarbazine is safe in dogs with HSA and prolongs TTM and survival time.     

Potency and stability of compounded cyclophosphamide: a pilot study

Compounding of drugs for use in veterinary oncology is becoming increasingly common. We obtained 15 mg cyclophosphamide capsules from five different compounding pharmacies and performed potency analyses at two time points, as well as stability at 60 days. Potency results for four out of five and zero out of five (4/10) samples analysed were inadequate. Stability at 60 days was acceptable for all but one sample. This pilot study raises several important points of concern when compounding chemotherapy in dogs and cats. Further studies are necessary to solidify this data. Collaboration between pharmacists, veterinarians and regulatory bodies is needed to ensure safe and accurate delivery of compounded drugs to client‐owned animals.     

mRNA差异显示法筛选环磷酰胺致突变相关基因的初步研究

为建立在药物致突变研究中应用银染mRNA差异显示的方法,试验提取未经过和经过环磷酰胺处理的BALB／e小鼠脾组织的总RNA,并以此为模板,采用dT12CG、dT22AG、dT12GG为锚定引物,通过反转录、差异显示PCR反应,6％变性聚丙烯酰胺凝胶电泳分离差异条带,回收后将其再扩增。对扩增条带进行克隆、测序。经Blast搜索,发现2条新的基因片段,为进一步研究致突变效应奠定了基础。     

环磷酰胺诱导黄羽肉鸡免疫抑制的研究

180只墟岗黄雏鸡随机分为4组，第1组为对照组，第2至4组为处理组，10日龄开始。对照组胸肌注射0．5mL的生理盐水，第2至4组分别按40、80和160mg／kg的剂量，同部位注射用生理盐水稀释的环磷酰胺(cyclophosphamide,CY)0．5mL；1次／d，连续注射3d．21、35、56日龄时每组随机抽取10只鸡进行屠宰取样；测定各组鸡的器官指数。另取3只鸡，测定脾脏淋巴细胞增殖转化和脾脏抗体形成细胞的数量。试验结果表明：一定剂量的环磷酰胺能显著降低墟岗黄鸡早期的体质量，且效果与注射剂量呈依赖关系；降低鸡早期脾脏指数和法氏囊指数；显著减少由ConA诱导的脾脏淋巴细胞的增殖转化；减少鸡早期脾脏抗体形成细胞的数量。     

The Effect of Cordyceps Militaris Polysaccharides on the Antioxidant Function in Mice Injected With Cyclophosphamide

To investigate the antioxidant effects of the Cordyceps militaris polysaccharides(CMP),90 male BALB/c mice were randomly divided into six groups, three experimental groups (CMP groups), model control group (CY group), blank control group (BC group) and positive control group (PC group). The mice in CMP and CY groups were given cyclophosphamide at 80 mg/kg via intraperitoneal injection for 3 d. Then the CMP groups were administrated 17.5, 35.0 and 70.0 mg/(kg·BW) CMP via gavage, respectively, BC and CY groups were given physiological saline, and PC group was orally administered 70 mg/(kg·BW) CMP, lasting 18 d. At 24 h after the last administration, heart, liver and kidney were collected to measure the levels of MAD, CAT, SOD, GSH-Px and T-AOC in the homogenate.SOD and TAOC activity of three CMP groups increased significantly (P<0.01), and the level of MDA decreased significantly (P<0.01) compared with CY group. As for CAT activity of heart, the value in low-dose CMP group had no significant change (P>0.05), while in middle and high-dose groups had significant (P<0.05) and extremely significant(P<0.01) increase respectively. Except that the hepatic GSH-Px activity increased significantly (P<0.05) in low-dose CMP group, the rest values of GSH-Px activities in all CMP groups increased extremely significantly (P<0.01). The results indicated that CMP could effectively improve the antioxidant function of mice and had the potential to be a good resource of new antioxidant drugs.     

鸡血藤总黄酮对环磷酰胺所致小鼠肝损伤的保护作用

目的 研究鸡血藤总黄酮对环磷酰胺所致小鼠肝损伤的保护作用。方法 将60只昆明系小鼠随机分为6组,每组10只,试验周期为7 d。对照组小鼠试验全程给予生理盐水。环磷酰胺（CTX）组小鼠1~4 d灌胃给予生理盐水;5~7 d腹腔注射80 mg/（kg·BW）CTX,每天给药1次。鸡血藤总黄酮（TFSD）100组、CTX+TFSD25组、CTX+TFSD50组、CTX+TFSD100组小鼠1~7 d分别灌胃给予100、25、50、100 mg/（kg·BW）TFSD,每天给药1次;试验5~7 d除TFSD100组小鼠腹腔注射生理盐水外,其余试验组腹腔注射80 mg/（kg·BW）CTX,每天给药1次。试验结束后对各组小鼠进行眼球采血及剖检;检测肝组织中超氧化物歧化酶（SOD）、髓过氧化物酶（MPO）、黄嘌呤氧化酶（XOD）以及谷胱甘肽过氧化物酶（GSH-Px）活力,测定血清中谷草转氨酶（AST）、谷丙转氨酶（ALT）和碱性磷酸酶（ALP）活力,检测血清中肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）和白介素-1β（IL-1β）含量。结果 环磷酰胺可显著（P<0.05）降低小鼠肝脏中SOD、GSH-Px以及血清中ALT活力,显著（P<0.05）升高肝脏中XOD与MPO活力、血清中ALP活力以及炎症细胞因子IL-1β和IL-6释放水平。25、50、100 mg/（kg·BW）的TFSD处理均可显著（P<0.05）抑制由CTX引起的SOD活力降低,25 mg/（kg·BW）的TFSD处理可显著（P<0.05）提高CTX处理小鼠的GSH-Px活力;50 mg/（kg·BW）的TFSD处理可显著（P<0.05）抑制由CTX引起的XOD和MPO活力升高,且可显著（P<0.05）降低CTX处理小鼠血清中的AST活力;25、50、100 mg/（kg·BW）的TFSD处理可显著（P<0.05）抑制由CTX引起的血清中ALP活力升高,同时降低ALT活力。25、50、100 mg/（kg·BW）的TFSD处理均可显著（P<0.05）抑制由CTX引起的IL-1β水平升高。结论 鸡血藤总黄酮可通过调节小鼠炎症因子释放以及肝组织中氧化还原酶水平对环磷酰胺所致肝损伤提供保护,推荐剂量为25 mg/（kg·BW）。