锥-46的耕牛血药浓度反相HPLC测定方法探索

用惠普ＨＰ１０５０四元泵、ＨＰ１０４０Ｍ二极管阵列检测器和色谱柱ＥｃｏｎｏｓｐｈｅｒｅＣ（１８）（２５０×４．６ｍｍ，５μ）探索了锥－４６的耕牛血药浓度测定条件。检测波长２５２±２ｎｍ，参比波长５５０±２ｎｍ。流动相为甲醇：乙腈：重蒸水＝２０：２５：５５（Ｖ／Ｖ），每３００ｍｌ加入二乙胺１０００μｌ，用磷酸调节至ｐＨ３．５，流速０．６ｍｌ／分。血浆中的锥－４６用磷酸酸化至ｐＨ２．４的甲醇溶液提取，平均提取效率达９９．３３±２．２８％。提取物在６０℃下减压浓缩。平均回收率为９９．４６±６．０７％。本法最低检测浓度为０．１μｇ／ｍｌ。在锥－４６浓度为０．１～１０μｇ／ｍｌ范围内线性良好（相关系数ｒ＝０．９９９９，峰面积＝０．６９２８＋１．５８１×锥－４６ｎｇ数）。锥－４６基本上能与血浆中的杂质成分完全分离。已应用于耕牛体锥－４６的药物代谢动力学的研究。     

以MTT比色法检测鸡脾淋巴细胞转化效果

本研究运用ＭＴＴ比色法以正交试验对鸡脾脏淋巴细胞转化的最佳条件（ｃｏｎＡ量、血清种类和浓度）进行了研究。结果表明，无论血清种类和浓度如何，低剂量ＣｏｎＡ（２．５～１０μｇ／ｍｌ）可以获得较好的转化效果。当ＣｏｎＡ的量确定时，低浓度鸡血清（０．５％～１．２５％）的转化效果优于高浓度（２％～３％）：５％犊牛血清优于任何浓度的鸡血清；无血清培养基优于血清培养基。其最佳组合为无血清ＲＰＭＩ１６４０培养基加ＣｏｎＡ２．５μｇ／ｍｌ。     

反相HPLC测定猪血浆中隐丹参酮及其活性代谢物的血药浓度

本试验建立了用反相ＨＰＬＣ同时进行猪血浆中隐丹参酮（ＣＴ）及其活性代谢物丹参酮ⅡＡ（Ｔｓ－ⅡＡ）的快速定量检测方法。采用ＹＷＧＣ１８键合相柱作为分析柱，甲醇－水（８５１５）为流动相，检测波长２５４ｎｍ，二苄基为内标。平均回收率分别为ＣＴ：８８．０９％，Ｔｓ－ⅡＡ：８８．５１％；原药及代谢物的检测限为１０ｎｇ，血浆中最低检出浓度为１６ｎｇ／ｍｌ。日内、日间的变异系数为１．５４％～６．２８％，血浆浓度０．０４～２．５μｇ／ｍｌ范围内呈线性关系，ｒ＝０．９９１４（ＣＴ）及０．９９８８（Ｔｓ－ⅡＡ）。用此法对５头猪进行了ｉｖＣＴ后的ＣＴ及Ｔｓ－ⅡＡ的血浆浓度测定，得出了药时曲线数据。     

克伦特罗在猪体内的生物利用度及药物动力学研究

６头体重４７．００±４．５７ｋｇ（Ｘ±Ｓ）的健康长白和大白杂交猪，拉丁方设计试验，按４ｍｇ／ｋｇ静注、肌注和内服克伦特罗，高效液相色谱法检测血浆中药物浓度，ＭＣＰＫＰ药代动力学程序处理药时数据。静注给药的药代动力学参数是：ｔ１／２α０．６２±０．１２ｈ，ｔ１／２β４．８７±１．５６ｈ，Ｖｄ（ａｒｅａ）４．４８±０．５６ｌ／ｋｇ，ＣｌＢ０．６３±０．１１ｌ／ｋｇ／ｈ，ＡＵＣ６．３９±１．２７μｇ／ｍｌ．ｈ。肌注给药的药动学参数是：ｔ１／２Ｋａ０．２２±０．１０ｈ，ｔ１／２α０．５６±０．２１ｈ，ｔ１／２β４．２５±１．１０ｈ，ｔｍａｘ０．６０±０．１３ｈ，Ｃｍａｘ１．２７±０．３５μｇ／ｍｌ，ＡＵＣ５．４８±１．２９μｇ／ｍｌ．ｈ，Ｆ８５．４０±４．６９％。内服给药的药动学参数是：ｔ１／２Ｋａ０．２８±０．１５ｈ，ｔ１／２Ｋｅ３．１５±０．３６ｈ，ｔｍａｘ１．４６±０．１９ｈ，Ｃｍａｘ０．６５±０．１３μｇ／ｍｌ，ＡＵＣ３．９３±０．９９ｍｇ／ｌ．ｈ，Ｆ６１．０２±１０．９０％。肌注给药的生物利用度与内服比较，差异极显著（Ｐ＜０．０１）。     

海南霉素的遗传毒性研究及其致癌性预测 1．海南霉素的体外致突变性研究

海南霉素是我国研制成功的第一个聚醚类抗生素，它具有良好的抗球虫作用。本文从遗传毒理学试验的生物学意义方面考虑，选择了２种体外致突变试验方法，对海南霉素的遗传毒性作了研究。１．体外哺乳动物细胞（ＣＨＬ）染色体畸变试验。根据海南霉素的细胞毒性试验结果，设立以下剂量组①６～１８ｈ方案：１．６４、８．２０和１６．４０μｇ／ｍｌ；②２４～Ｏ和４８～０ｈ方案：０．０９５、０．１９０和０．９５０μｇ／ｍｌ。同时设溶剂和阳性对照。试验结果经Ｘ２检验为阴性。２．体外哺乳动物细胞（ＣＨＬ）姊妹染色单体交换试验，根据海南霉素的细胞毒性试验结果，设立以下剂量组：０．２、２．０、２０．０和３３．０μｇ／ｍ１。同时设溶剂和阳性对照。试验结果经ｔ检验为阴性。     

猪流行性腹泻病毒适应Vero细胞培养及以传代细胞毒制备氢氧化铝灭活疫苗免疫效力试验

采用在病毒培养液中加５～１０μｇ／ｍｌ胰酶的培养方法，将猪流行性腹泻病毒（ＰＥＤＶ）ＣＶ＿（７７７）适应于Ｖｅｒｏ细胞，并传４５代．细胞病变规律．经免疫荧光检查阳性，电镜观察可见典型冠状病毒粒子，猪传染性胃肠炎（ＴＧＥ）免疾荧光检查阴性，猪流行性腹泻（ＰＥＤ）血清可抑制细胞病变（ＣＰＥ）．ＰＥＤＶＣＶ＿（７７７）毒株１１、２５、２８、４０及４４代传代毒的毒价分别为１０￣（３．５）、１０￣（５．５）、１０￣（６．５）、１０￣（７．０）和１０￣（７．０）ＴＣＩＤ＿（５０）／０．３ｍｌ。以１１、２１及２２代的毒１０ｍｌ头口服接种未吃初乳仔猪，可使之典型发病，免疫荧光及电镜观察均呈阳性．分别以２５、２８及３１代毒０．５ｍｌ／头、１ｍｌ／头、２ｍｌ／头口服接种３日龄仔猪１８头．除０．５ｍｌ组有１头反应外，均未发病，攻毒试验的总保护率为８７．５％，对照组１００％发病．以２８代毒制备氢氧化铝灭活苗，后海穴位接种，主动免疫组８５．１９％保护，被动免疫组８５％保护，对照组１００％及９２．３％发病．     

分光光度法测定盐酸噻咪唑片的含量

以０．１ｍｏｌ／Ｌ的盐酸液为溶剂,采用分光光度法测定盐酸噻咪唑片的含量,测定浓度在２～１０μｇ／ｍｌ范围内线性关系良好（ｒ＝０．９９９９）,平均回收率为９９．６％,ＲＳＤ为０．４４％,方法简便,快速、准确,重现性好。     

多杀巴氏杆菌的一种选择性分离培养基

以马丁培养基为基础，加入促生长因子牛全血和抑菌剂硫酸新霉素和盐酸洁霉素制成多杀巴氏杆菌分离培养基．当培养基中硫酸新霉素的浓度低于２．５μｇ／ｍｌ时，对多杀巴氏杆菌生长无影响；浓度为２．５μｇ／ｍｌ时．对其个别菌株的生长稍有抑制作用；当浓度增大到到５μｇ／ｍｌ时，荚膜Ａ型菌株的生长明显受到抑制，Ｂ型菌株部分受到抑制，而Ｄ型菌株基本不受影响；当浓度达到１０μｇ／ｍｌ时，各型菌株均不能生长．当培养基中含硫酸新霉素２．５μｇ／ｍｌ和盐酸洁霉素１．０μｇ／ｍｌ时，仅个别菌株的生长稍受抑制，而绝大多数菌株生长不受影响．当培养基中含硫酸新霉素２．０μｇ／ｍｌ和盐酸洁霉素１．０μｇ／ｍｌ时．对各型多杀巴氏杆菌的生长均无抑制作用，并可抑制环境中绝大多数杂菌的生长。因此，可以作为选择性培养基分离多杀巴氏杆菌。     

高浓度铁剂对哺乳仔猪生长发育的影响

采用随机单位组设计，将１６窝的９６头哺乳仔猪分为３组，每组３２头。试验１组、２组于３日龄分别注射１ｍｌ自制的高浓度铁剂“铁血素１号”（含铁１５０ｍｇ／ｍｌ）、“铁血素２号”（含铁１００ｍｇ／ｍｌ），３组为空白对照。试验结果：３５日龄断奶时，１组和２组的日增重分别比对照组高３６．７６％和３２．３５％（Ｐ＜０．０１）；３日龄时，每１００ｍｌ血液中血红蛋白含量，各组均在９～１０ｇ（Ｐ＞０．０５），到１２、２５、３５日龄时，１组和２组均在１２～１４ｇ，与对照组的６～９ｇ相比差异极显著（Ｐ＜０．０１）；１组、２组的经济效益分别比对照组高３６．１２％和３１．９０％。     

鸡静注,肌注及内服氟哌酸的药物动力学研究

选用４２只健康ＡＡ鸡，分３组（每组１４只）进行药物动力学研究：静注、肌注及内服的剂量均为１０ｍｇ／ｋｇ。用二氯甲烷提取血浆中的药物，反相高效液相色谱法测定血浆氟哌酸的浓度，测定时以吡哌酸为内标。所得药物一时间数据用ＭＣＰＫＰ计算机程序处理，静注给药拟合二室开放模型，主要动力学参数是：ｔ１／２α０．１４±０．０８ｈ，ｔ１／２β３．６５±１．０３ｈ，Ｋｅ０．８８±０．４０ｈ－１，Ｖｄ３．８１±０．９８Ｌ／ｋｇ，ＡＵＣ１４．１４±３．２９μｇ／ｍｌ．ｈ。内服和肌注给药均适合具有一级吸收二室开放模型，肌注的主要动力学参数是：ｔ１／２ｋａ０．２２±０．１０ｈ，ｔ１／２α１．０３±０．６５ｈ，ｔ１／２β６．８７±２．８６ｈ，Ｋｅ０．２８±０．１０ｈ－１，ＡＵＣ９．８６±２．８８μｇ／ｍｌ．ｈ，Ｆ６９．７８％。内服的主要动力学参数是：ｔ１／２ｋａ０．６９±０．２３ｈ，ｔ１／２α１．７６±０．８９ｈ，ｔ１／２β１０．４３±３．２２ｈ，Ｋｅ０．２１±０．０５ｈ－１，ＡＵＣ８．５６±１．３６μｇ／ｍｌ．ｈ，Ｆ６０．５２％。鸡静注、肌注、内服氟哌酸的药动学特征是吸收较快，在体内分布广泛、半衰期较长。     

Heartworm in dogs in Canada in 1985

In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).

Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada.

     

Heartworm in dogs in Canada in 1988

In late November 1988, 1581 small and mixed animal clinics and institutional veterinarians across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1988, and 46% of them responded. Veterinarians reported that 181,577 dogs were blood-tested for heartworm disease and 367 dogs were found with D. immitis microfilariae. Another 60 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1988 as 441 (0.24%).     

Heartworm in dogs in Canada in 1984

In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.

Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection.

     

Heartworm in dogs in Canada in 1989

In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.

Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively.

     

Heartworm in dogs in Canada in 1987

In late November 1987, 1246 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1987, and 50% of them responded. Veterinarians reported that 165,428 dogs were blood tested for heartworm disease and 511 dogs were found with D. immitis microfilariae. Another 78 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1987 as 589 (0.35%).     

Heartworm in dogs in Canada in 1986

In late December 1986, 1224 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1986; 46% of them responded. Veterinarians reported that 150,989 dogs were blood-tested for microfilariae and 869 dogs were found with heartworm. Another 65 dogs were amicrofilaremic but diagnosed with heartworm disease and one was found with heartworm at necropsy to give the total number diagnosed in 1986 as 935 (0.62%).

Heartworm was reported from Manitoba, New Brunswick, Ontario and Quebec, but most (810) of the cases were from Ontario. South-western Ontario continued to be the primary focus of the infection in Canada. There were 103 cases reported from Quebec, mostly from and around Montreal, and 21 cases from Manitoba, from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 33% of the cases were observed with clinical signs of heartworm disease and 81% had a history of not having left Canada.

     

Heartworm in dogs in Canada in 1983

In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.

Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously.