银翘天甘醇提物含药血清制备方法的研究

为探索银翘天甘醇提物含药血清的制备工艺,采用单因素试验优化采血时间、给药次数及给药剂量,于277nm检测小鼠血清中连翘苷含量,经中心组合设计与响应面分析获得银翘天甘含药血清最佳制备工艺条件。结果表明,制备银翘天甘含药血清最佳工艺条件是以0.01g/mL银翘天甘醇提物给药4次,于最后一次给药后31min采血获得的含药血清中连翘苷含量最高,达到193.788μg/mL。通过验证试验,此模型确认有效。     

银翘天甘复方制剂中绿原酸的测定

本实验旨在探索建立银翘天甘复方制剂中绿原酸含量的检测方法.采用紫外分光光度法(UV),检测波长为327nm;薄层色谱法(TLC),展开剂为乙酸丁酯-甲酸-水(7.5:2.5:2.5).结果表明:绿原酸浓度为2.5～15μg/mL时有较好的线性关系,回归方程:A=0.1897C-0.0033,r=0.9997(n=5),回收率为99.4%,RSD为0.2%.薄层色谱能将复方制剂中的绿原酸很好的分离出来,样品中的绿原酸比移值(Rf值)=0.28±0.01(n=5),与对照品之间的Rf值基本一致.银翘天甘复方制荆中舍有绿原酸,且UV、TLC可作为银翘天甘复方制剂中绿原酸的定性分析检测方法.     

响应面法优化杜仲叶提取物的制备工艺

优化杜仲叶提取物制备的工艺条件,在单因素试验基础上,以杜仲叶提取物浸膏得率和绿原酸转移率为评价指标,采用Box-Behnken中心组合设计法优化杜仲叶提取物制备的工艺条件。杜仲叶提取物制备的最佳工艺条件为乙醇浓度500mL/L,液料比10∶1(mL/g),提取温度97℃,提取4次,每次30min;在此最佳工艺条件下,杜仲叶提取物浸膏得率和绿原酸转移率综合评分均值为99.08,与预测值基本一致。响应面法(RSM)优化的杜仲叶提取物制备工艺稳定可靠。     

桃金娘水提物对小鼠经口给药急性和亚急性毒性研究

研究旨在评价桃金娘水提物经口给药的安全性,为其临床应用提供参考依据。将SPF级KM小鼠随机分成空白组和给药组,经口给药进行急性毒性试验以单次最大灌胃量(0.04 mL/g)给予桃金娘水提物(200 g/kg),连续给药3次,每次间隔4 h。连续观察给药后14 d内小鼠的生存状况和不良反应,监测其体重、摄食量,并详细记录。亚急性毒性试验将给药组分为桃金娘水提物高、中、低剂量(9.1、4.55、2.27 g/kg)组,每天给药1次,重复经口给药28 d,观察记录其生理状况、体重变化及摄食量。每次试验结束时,采集血清进行生化分析,观察心、肝、脾、肺、肾的病理变化,计算脏器指数。结果表明,在急性经口给药试验中,未观察到与治疗相关的中毒和死亡迹象;桃金娘水提物单日剂量在600 g/kg以下,对小鼠具有良好的耐受性,耐受量是人体剂量的310倍;亚急性试验表明,给药组小鼠的摄食量、每周体重增加百分比、各脏器指数、血清肝功能和肾功能指标与空白组比较均无显著差异(P0.05)。研究表明,桃金娘水提物未引起小鼠的急性和亚急性反应,安全性高,在人体规定治疗剂量下使用具有安全可靠性。     

金荞麦根提取物对鸡脾淋巴细胞活性影响的血清药理学研究

应用中药血清药理学的方法探讨了金荞麦根乙醇提取物药物血清对鸡脾T、B淋巴细胞增殖活性影响的时效及量效关系。首先通过预试验确定含药血清制备方案,包括给药剂量、给药方案和采血时间点的确定。根据预试验结果,设立4个试验组即高(40g生药/kg.d)、中(24g生药/kg.d)、低剂量组(6g生药/kg.d)和空白对照组(灌胃等体积的生理盐水),每天灌服2次,连续14d,末次灌服后40、80、120、180min分别翅下静脉采血,无菌分离血清,经56℃、30min灭活备用。同时,通过体外试验系统中的血清不同添加量试验,确定以2.5%的金荞麦根含药血清为体外细胞培养体系中含药血清的添加量,采用MTT法来探讨金荞麦根各含药血清对鸡脾淋巴细胞增殖活性的影响。结果表明,金荞麦根含药血清对鸡脾淋巴细胞的增殖活性呈明显的时-效及量-效关系,其中以中剂量组120min时相含药血清对鸡脾淋巴细胞的增殖活性为最强(P0.05)。     

桃金娘水提物抗肝脏纤维化作用的试验研究

本试验旨在研究桃金娘水提物对CCl₄复合因素造模法诱导的肝脏纤维化大鼠的保护作用,探讨其作用机制。取SD大鼠72只,随机分为模型组、秋水仙碱组（1×10^-4 mg/g）、桃金娘水提物低剂量（1.5 mg/g）、中剂量（3 mg/g）、高剂量（6 mg/g）组及正常组,每组12只。除正常组外,其余各组大鼠于第1~4周皮下注射40% CCl₄花生油溶液,注射剂量为0.3 mL/100 g体重,隔日1次,连续4周,同时第2~4周给予30%乙醇灌胃1 mL/只,隔日1次,连续3周。各组大鼠于第5周开始灌胃给药,灌胃体积量为1 mL/100 g体重,正常组、模型组给予等量生理盐水,桃金娘水提物组分别按照相应剂量灌胃给药,每日1次,连续给药4周,给药的同时仍继续造模。给药结束后,检测各组大鼠血清中丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）活性、总胆红素（TBil）、α-平滑肌肌动蛋白（α-SMA）、转化生长因子-β（TGF-β）等的水平、丙二醛（MDA）含量及超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）活性;观察肝脏组织HE及Masson染色的变化。试验结果显示,桃金娘水提物可显著或极显著降低肝脏纤维化大鼠AST、ALT活性、TBil、TGF-β及α-SMA水平（P<0.05;P<0.01）,提高大鼠血清SOD、GSH-Px活性（P<0.05）,降低MDA含量（P<0.01;P<0.05）,减轻肝脏纤维化大鼠肝脏病理损伤程度。以上结果表明,桃金娘水提物对CCl₄复合因素所致肝脏纤维化大鼠的肝脏功能具有明显的恢复作用,可增强其抗氧化能力,减轻肝细胞损伤,具有较好的抗肝脏纤维化作用。     

玉米须水提物的利尿作用及抗菌活性研究

为简化玉米须药用成分的加工提取方法,降低成本,并使其能够更加合理、有效发挥其药理作用,试验采用水煎浸提法从玉米须中提取多糖成分,并以小白鼠为研究对象,探究玉米须水提物(主要为多糖)的利尿效果;采用回流法和滤纸片法进行抑菌性研究。结果表明:与阴性对照组比较,3种给药途径差异显著(P0.05),对小白鼠均有利尿效果;在大肠杆菌和金黄色葡萄球菌的培养基中,玉米须水提物在浓度为0.1 g/mL以上时所形成的抑菌圈直径,与阴性对照组比较差异极显著(P0.01)。说明玉米须水提物腹腔注射给药有很好的利尿作用,皮下注射与灌胃给药次之;在本试验浓度范围内,0.1 g/mL浓度以上的玉米须水提物对金黄色葡萄球菌和大肠杆菌有抑菌效果,且对金黄色葡萄球菌的抑菌效果较好。     

银翘口服液制备工艺研究

目的:本研究旨在确定银翘口服液的制备工艺。方法:采用L_9(3~4)正交试验设计表,以挥发油的提取量和水提液中绿原酸含量作为指标,分别对挥发油提取和水提工艺进行优化。结果:挥发油提取工艺为加5倍水,浸泡0.5 h,提取5 h;水提工艺为加5倍水煎煮,每次煎煮1.5 h,煎煮3次。结论:本研究确定的生产工艺稳定可靠。     

葛根素对摘除卵巢小鼠血清激素含量的影响

目的:阐明葛根素对摘除卵巢小鼠血清中几种激素含量的影响。方法:40只21日龄雌性小鼠建立摘除卵巢模型,将其分为阴性对照组、试验组(葛根素组)、阳性对照组Ⅰ(己烯雌酚组)和阳性对照组Ⅱ(己烯雌酚+孕酮混合组),另外设正常对照组、每组10只,共50只小鼠。正常对照组和阴性对照组蒸馏水灌胃;试验组葛根素水溶液(1 mg/mL)灌胃;阳性对照组Ⅰ己烯雌酚悬浮液(20μg/mL)灌胃;阳性对照组Ⅱ己烯雌酚悬浮液(20μg/mL)和黄体酮注射液(20 mg/mL)混合灌胃。隔日给药一次,各组每次每只0.1 mL,连续给药3周。给药结束后采血,采用放射免疫法测定小鼠血清中激素含量。结论:摘除卵巢小鼠血清中雌二醇、孕酮含量显著降低,生长激素含量显著升高,而血清催乳素含量无明显变化。葛根素对摘除卵巢小鼠血清雌二醇、催乳素含量无明显影响,但能够影响摘除卵巢后血清中孕酮含量,使之基本恢复到摘除卵巢前水平,并能显著抑制卵巢摘除所引起的血清生长激素含量过高,并使其恢复到正常水平。     

绿原酸在家兔体内的药动学研究

建立了血浆中绿原酸的HPLC分析方法,比较了金银花三种注射液中绿原酸在家兔体内的药动学差异,旨在为研究金银花连翘药提供实验数据。将18只健康无伤的家兔随机分成三组,按0.5 mL/kg单次肌肉注射给药,HPLC同时测定不同时间点血浆中绿原酸浓度。用MCPKP程序计算动力学参数。健康家兔肌注金银花单提注射液(A),金银花、连翘混合提取注射液(B),金银花、连翘单提合并注射液(C)后,绿原酸在家兔体内的药动学过程均符合一级吸收二室模型。B、C注射液中绿原酸的Tmax、Tα/2、T1/2ka与A相比呈差异显著性,而Tβ/2差异不显著,表明金银花配伍连翘后能促进绿原酸在体内吸收分布,迅速达到血峰浓度,对消除半衰期的影响不大。B与C中绿原酸的药动学参数差异不显著,表明连翘与金银花配伍时,不管是混提还是单提后合并,对绿原酸在家兔体内的药动学特征无明显影响。     

基于转录组学技术分析绿原酸对鸡源大肠杆菌抑菌及耐药消除的作用机制

为探究绿原酸对鸡源抗生素耐药大肠杆菌抑菌及耐药消除的作用机制，以影印法分离绿原酸浓度为1.25 mg/mL（亚抑菌浓度，1/2 MIC）的LB肉汤培养的第3代禽源大肠杆菌耐药逆转菌株，以微板法测定耐药逆转菌株的左氧氟沙星最小抑菌浓度（MIC），并进一步通过转录组测序方法分析绿原酸消除大肠杆菌耐药性的分子机制。结果显示：耐药逆转菌株对左氧氟沙星的MIC由16 μg/mL降低至4~8 μg/mL，说明1.25 mg/mL的绿原酸可在一定程度上消除耐药大肠杆菌的左氧氟沙星耐药性。为进一步解析绿原酸对鸡源大肠杆菌耐药消除作用的分子机制，通过转录组测序方法对耐药性消除前后鸡源大肠杆菌基因表达水平进行对比分析发现：绿原酸作用后共有12个基因的表达量发生显著下调，通过荧光定量PCR验证结果基本一致。经过GO功能富集分析和KEGG代谢通路富集分析发现，差异表达基因在功能上集中于膜组成成分和DNA重组，在代谢通路富集中差异基因均与代谢相关，差异基因中bhsA、cysP为Coli-HB染色质转录RNA（GenBank登录号：CP020933），oqxA、oqxR、emaA、pinR、xerD、folA、repA、repC、adhP、IS26为Coli-HB质粒1转录RNA（GenBank登录号：CP020934）。从差异表达基因的分布可以看出，绿原酸可以抑制细菌DNA重组、使细菌抗逆性减弱、抑制耐药基因活性等，在一定程度上消除大肠杆菌的左氧氟沙星耐药性，起到抑菌及耐药消除作用。     

Florfenicol pharmacokinetics in lactating cows after intravenous, intramuscular and intramammary administration

Pharmacokinetics of florfenicol 30% injectable solution was determined in lactating cows after intravenous, intramammary and intramuscular administration. Serum concentration-time data generated in the present study were analysed by non-compartmental methods based on statistical moment theory. Florfenicol half-life was 176 min, mean residence time 129 min, volume of distribution at steady-state 0.35 L/kg, and total body clearance 2.7 mL/min·kg after intravenous administration at 20 mg/kg. The absorption after intramuscular administration appeared slow and the kinetic parameters and the serum concentration vs. time curve were characteristic of absorption rate-dependent elimination. The absorption after intramammary administration of florfenicol at 20 mg/kg was good (53.9%) and resulted in serum concentrations with apparent clinical significance. The intramammary administration resulted in serum florfenicol concentrations that were significantly higher than the respective serum concentrations following Intravenous administration 4 h after administration and thereafter. Florfenicol absorption was faster from the mammary gland than from the muscle. The maximum serum concentrations ( C _max) were 6.9 μg/mL at 360 min after intramammary administration and 2.3 μg/mL at 180 min after intramuscular administration. The bioavailability of florfenicol was 54% and 38% after intramammary and intramuscular administration, respectively. The C _max in milk was 5.4 μg/mL at 180 min after intravenous and 1.6 μg/mL at 600 min after intramuscular administration.     

Pharmacokinetic Disposition of Cefazolin in Serum and Tissue during Canine Total Hip Replacement

Intraoperative cefazolin concentrations were measured in serum, joint capsule, cancellous bone of the acetabulum, and proximal cancellous bone of the femur in 15 dogs undergoing total hip replacement. Cefazolin (22 mg/kg intravenously [IV]) was administered every hour for three doses. The mean peak serum concentrations (+/- SEM) were 387.79 +/- 27.56 micrograms/mL, 521.71 +/- 28.00 micrograms/mL, and 542.20 +/- 30.91 micrograms/mL, respectively. Mean serum concentrations just before administration of doses 2 and 3 were 51.77 +/- 2.39 micrograms/mL, and 64.84 +/- 3.46 micrograms/mL, respectively. The mean cefazolin concentrations in the joint capsule, cancellous bone of the acetabulum, and cancellous bone of the femur were 34.71 +/- 2.50 micrograms/g, 28.70 +/- 7.40 micrograms/g, and 36.20 +/- 3.80 micrograms/g, respectively. The minimum inhibitory concentration of cefazolin for 90% of the common contaminants (MIC90) in this clinic is less than or equal to 2 micrograms/mL or per gram of tissue. Serum concentrations never fell below 15 times the MIC90 (lowest trough, 35.93 micrograms/mL), and the lowest tissue concentration (6.57 micrograms/mL in cancellous bone from the acetabulum) was still more than 3 times the MIC90. The mean tissue concentration was 15 times the MIC90.     

芩花注射剂的制备工艺及主药含量测定

芩花注射剂主要由黄芩、金银花、百部等组成。采取单因素试验筛选药物提取次数、提取时N／次、料液比,通过正交试验以绿原酸提取量为依据最后确定提取次数为2次、每次提取1h、料液比为1：10。采用HPLC法测定该注射液中黄芩苷和绿原酸含量,以峰面积（Y）对进样浓度（X）做线性回归,黄芩苷回归方程：Y=0．04X一14．78（r。=0．999）,绿原酸回归方程：Y=0．04X一10．73（r2=0．998）,精密度、回收率、重复性、稳定性等指标均符合含量测定要求。本方法准确度好且操作简便。经测定该注射液中黄芩苷和绿原酸的含量分别为3．292、0．860mg／mL。     

Pharmacokinetics and renal clearance of oxytetracycline in piglets following intravenous and oral administration

Summary

The pharmacokinetics of oxytetracycline (OTC) in three weaned piglets was studied following three routes of administration: intravenously, orally as drench, both at a dose of 20 mg/kg, and orally as medicated (400 ppm OTC) pelleted feed administered during 3 consecutive days. Analysis of the intravenous data according to the three compartment pharmacokinetic model revealed that OTC was well distributed in the body (Vie 1.621/kg), had an overall body clearance of 0.25 litre/kg/h, and the elimination half‐lives were in the range between 11.6 and 17.2 hrs.

The mean OTC binding to plasma proteins was 75.5 ± 4%. Following the drench route of administration the maximum plasma OTC concentration was achieved between 1 and 5 h post application and ranged between 1.18 and 1.41 μg/ml. The mean maximum plasma OTC concentration during medicated feed administration was 0.20 ± 0.06 μg/ml, which was achieved approximately 30 hours after the onset of the administration. A steady state OTC plasma level (approximately 0.2 μg/ml) was maintained till the end of the trial. Within 48 hours after cessation of medicated feed administration the plasma OTC levels were beneath 0.06 μg/ml. The mean OTC bioavailabilities of the oral routes were low: after the drench route of administration 9.0 ± 0.67%, and after medicated pelleted feed administration 3.69 ± 0.8%.

The mean OTC renal clearances of each piglet ranged between 10.1 and 13.9 ml/min/kg (based on free OTC plasma fractions). The renal OTC clearance values were urine flow dependent in all piglets and significantly correlated with the renal creatinine clearance (P< 0.005), being 3–5 times higher than the latter. It is concluded that in piglets OTC is excreted mainly by glomerular filtration and partly by tubular secretion. The potential clinical efficacy of 400 ppm OTC as medicated feed with respect to treatment, e.g. atrophic rhinitis, is discussed.     

复方伊维菌素乳液在山羊体内的药代动力学研究

为了探讨复方伊维菌素乳液在动物体内的药物代谢,为兽医临床提供用药参考。试验选取7只山羊,每只山羊按0.1 mL/kg (伊维菌素0.2 mg/kg, 阿苯达唑10 mg/kg)剂量口服,给药后0.5、1、2、3、4、6、8、12、16、24、36、48、60 h颈静脉采血5 mL,分离血清,-20 ℃保存,用高效液相色谱仪检测样品血药浓度。试验结果表明,①伊维菌素在山羊体内的代谢情况为：0.5 h,0.112151 μg/mL; 第1次达峰时间为4 h, 0.302702 μg/mL;第2次达峰时间为16 h,0.258284 μg/mL;60 h,0.011118 μg/mL。②阿苯达唑在山羊体内的代谢情况为：0.5 h, 0.049285 μg/mL;第1次达峰时间为8 h,4.95283 μg/mL ;第2次达峰时间为16 h,5.694551 μg/mL;60 h,0.06434 μg/mL。复方伊维菌素中的伊维菌素和阿苯达唑在山羊体内代谢时间短,第60小时已达到很低的血药浓度。     

HPLC法同时测定金叶清瘟散中绿原酸和咖啡酸的含量

为了建立一种同时测定金叶清瘟散中绿原酸和咖啡酸的高效液相色谱法,选用Agilent C18色谱柱（250 mm×4.6 mm,5μm）,乙腈∶0.4%磷酸溶液（12∶88）等度洗脱,进样量10μL,流速1 mL/min,波长为328 nm进行检测。结果显示,被测物质能与样品中杂质有效分离,绿原酸和咖啡酸分别在5.0~500μg/mL和2.5~250μg/mL的范围内呈良好线性关系,两种药物的添加回收率均高于95%,RSD小于2%。该方法简便、快速、准确,适用于金叶清瘟散中绿原酸和咖啡酸的同时检测。