Simultaneous determination of doxorubicin and its dipeptide prodrug in mice plasma by HPLC with fluorescence detection

A simple and sensitive high performance liquid chromatography with fluorescence detection(HPLC–FD)has been developed for simultaneous quantification of doxorubicin(DOX) and its dipeptide conjugate prodrug(PDOX) in mice plasma.The chromatographic separation was carried out on an Amethyst C_(18)–H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min.The excitation and emission wavelengths were set at 490 and 550 nm,respectively.The method was comprehensively validated.The limits of detection were low up to 5.0 ng/m L for DOX and 25.0 ng/m L for PDOX.And the limits of quantification were low up to 12.5 ng/m L for DOX and 50 ng/m L for PDOX,which were lower than those for most of the current methods.The calibration curves showed good linearity(R^2>0.999) over the concentration ranges.The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX.Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%.The results show that the developed HPLC–FD method is accurate,reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

Non-thermal Plasma Suppresses Bacterial Colonization on Skin Wound and Promotes Wound Healing in Mice

The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice.Two 6-mm wounds along the both sides of the spine were created on the back of each mouse(n=80) by using a punch biopsy.The mice were assigned randomly into two groups,with 40 animals in each group:a non-thermal plasma group in which the mice were treated with the non-thermal plasma;a control group in which the mice were left to heal naturally.Wound healing was evaluated on postoperative days(POD) 4,7,10 and 14(n=5 per group in each POD) by percentage of wound closure.The mice was euthanized on POD 1,4,7,10,14,21,28 and 35(n=1 in each POD).The wounds were removed,routinely fixed,paraffin-embedded,sectioned and HE-stained.A modified scoring system was used to evaluate the wounds.The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group,earlier than in control group in which acute inflammation reached a peak on POD 7,and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7(P<0.05).The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group(P<0.05).The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10(P<0.05 for all).The count of bacterial colonies was 103 CFU/mL on POD 4 and <20 CFU/mL on POD 7,significantly lower than that in control group(109 CFU/mL on POD 4 and >1012 CFU/mL on the POD 7)(P<0.05).It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colonization.

Systemic study on the safety of immuno-deficient nude mice treated by atmospheric plasma-activated water

Cold atmospheric-pressure plasma is a new technology, widely used in many fields of biomedicine,especially in cancer treatment. Cold plasma can selectively kill a variety of tumor cells, and its biological safety in clinical trials is also very important. In many cases, the patient's immune level is relatively low, so we first studied the safety assessment of plasma treatment in an immunocompromised animal model. In this study, we examined the safety of immuno-deficient nude mice by oral lavage treatment of plasma-activated water, and studied the growth status, main organs and blood biochemical indexes. Acute toxicity test results showed that the maximum dose of plasma treatment for 15 min had no lethal effect and other acute toxicity. There were no significant changes in body weight and survival status of mice after 2 min and 4 min of plasma-activated water(PAW)treatment for 2 weeks. After treatment, the major organs, including heart, liver, spleen, lung and kidney, were not significantly changed in organ coefficient and tissue structure. Blood biochemical markers showed that blood neutrophils and mononuclear cells were slightly increased, and the others remained unchanged. Liver function, renal function, electrolytes, glucose metabolism and lipid metabolism were not affected by different doses of PAW treatment. The above results indicate that PAW treatment can be used to treat immuno-deficient nude mice without significant safety problems.

Study on changes of polyamine levels in mice with the development of U14 cervical cancer

This study was performed to investigate the possible involvement of polyamines in the development of cervical cancer. The objective of the present study was therefore to find the specific polyamine indicators, which could be used as useful markers for the early determination of cervical cancer. A simple method for the simultaneous determination of plasma concentrations of five polyamines in normal and U14 model mice was developed by using HPLC-MS. The samples were derivatized by benzoyl chloride. The derived polyamines were separated on a C18 column by a gradient elution with methanol-water, and then detected with HPLC-MS. The results showed that all polyamine levels in the U14 model mice were higher than those in normal ones. The cadaverine, putrescine and 1, 3-diaminopropane levels were significantly higher in U14 model mice plasma than those in normal mice plasma, especially the putrescine and 1, 3-diaminopropane (Po0.01). The cadaverine, putrescine and 1, 3-diaminopropane levels were significantly higher in U14 mice model urine than those in normal mice urine, especially the cadaverine and 1, 3-diaminopropane (Po0.01). Putrescine, cadaverine and 1, 3-diaminopropane might be the indicators of the cervical cancer.

Effects of Zaizhang－Ⅰ，a Traditional Chinese Medicine，on Immunologically Mediated Aplastic Anemia in Mice

(周剑锋)(何美冬)(李鸣真)(杨炜)(唐锦治)(沈关心)(朱慧芬)(邵静芳)(杨敬)ＥｆｆｅｃｔｓｏｆＺａｉｚｈａｎｇ－Ⅰ，ａＴｒａｄｉｔｉｏｎａｌＣｈｉｎｅｓｅＭｅｄｉｃｉｎｅ，ｏｎＩｍｍｕｎｏｌｏｇｉｃａｌｌｙＭｅｄｉａｔｅｄＡｐｌａｓｔｉｃＡｎｅｍｉａｉｎＭｉ...

Comparative Assessment of Melatonin-Afforded Protection in Liver, Kidney and Heart of Male Mice against Doxorubicin Induced Toxicity

Melatonin (MEL) was investigated for protection against the anthracycline antibiotic doxorubicin (Dox) that is well known for its oxidative damage to various body organs. It was aimed to have a comparison of this protection to heart, liver and kidney in the treated subjects. In this study, groups of mice were treated with Dox and melatonin and their individual or combined effects were evaluated by assessing lipidperoxidation, non-protein sulfhydryls (NP-SH) and nitrate/nitrite (NO) contents in these tissues. Plasma aminotransferases, LDH and CK-MB enzyme activities were measured. Moreover, these tissues were subject to histopathological assessment. MEL co-treatment significantly prevented any rise in lipidperoxides more significantly in heart and liver as compared to kidney. In tandem, MEL prevented a decline in GSH that was observed by Dox alone in liver and kidney. Dox significantly increased total NO levels in all the tissues. Melatonin at both dose levels could not afford protection against nitrosative stress. MEL in combination treatment provided significant

2型糖尿病db/db小鼠骨强度和肠道内容物菌群的变化

目的观察db/db小鼠血糖、骨强度和肠道菌群的变化。方法20周龄雄性db/db小鼠及野生型小鼠各6只,检测空腹血糖,行三点弯曲实验检测股骨力学指标,采用16S rRNA高通量测序技术对小鼠肠道内容物基因V3-V4区进行测序及生物信息分析。结果db/db组小鼠体重(P<0.01)和血糖(P<0.05)均显著升高。db/db小鼠胫骨刚度、最大位移、最大能量吸收、最大载荷均显著低于WT小鼠(P<0.01)。db/db小鼠α多样性指标中的Chao1指数、ACE指数和observed species均显著降低(P<0.05);两者肠道细菌群落组成存在显著差异(ANOSIM:P=0.02)。db/db小鼠毛螺菌属(Lachnospira)显著降低(P<0.01);变形菌门(Proteobacteria)、δ-变形菌纲(Deltaproteobacteria)、脱硫弧菌目(Desulfovibrionales)显著升高(P<0.05),Alloprevotella属极显著升高(P<0.01)。结论本研究结果表明db/db小鼠高血糖且骨强度降低,是T2DOP的适宜动物模型。db/db小鼠与WT小鼠肠道菌群的多样性及特定菌群丰度存在差异,为研究肠道菌群与糖尿病性骨质疏松提供了理论基础。

年轻血浆抗衰老作用及其机制研究进展

随着年龄的增长,人们患有糖尿病、心血管疾病、癌症和神经退行性疾病的风险增加,年轻血浆中的有关成分对于人类寿命、疾病治疗和预防作用已成为研究的新领域。本文就年轻血液在小鼠中应用后对年老小鼠脑神经、血管、胰腺、肝脏、骨的影响及其相关作用机制的研究进行综述,以此为今后年轻血浆在人体中的抗衰老研究提供参考价值。

HPLC-MS法测定小鼠血浆中蒜氨酸的浓度

目的:建立测定小鼠血浆中蒜氨酸浓度的液相色谱-串联质谱法。方法:使用大气压化学电离源(APCI),在正离子条件下选择性离子监测(SIM)。用于定量分析的离子分别为m/z179→m/z200(蒜氨酸)和m/z195→m/z217(内标,咖啡因)。结果:蒜氨酸浓度与峰面积呈良好的线性关系,线性范围为1.35～12.15 μg/ml。其最低检测限(S/N=3)可达1.62 pg/μl。结论:HPLC-MS法灵敏、快速、准确,可用于小鼠血浆中蒜氨酸的浓度测定及临床前药动学研究。

GC-MS法同时测定小鼠血浆中冰片和丹皮酚浓度

目的:建立测定小鼠血浆中冰片和丹皮酚的气相色谱-质谱(GC-MS)定量分析方法。方法:血浆样品经环己烷提取,采用 GC-MS 选择性离子和二级质谱分别监测冰片和丹皮酚血药浓度。结果:本法可同时测定血浆中2种药物浓度,冰片在5ng·mL^(-1)～10μg·mL(-1),丹皮酚在50ng·mL^(-1)～10μg·mL(-1)范围内,峰面积与其浓度呈良好的线性关系;最低检测限分别为2.5ng·mL^(-1),25ng·mL^(-1);日内 RSD 分别为4.4%～8.4%,5.0%～5.5%;日间 RSD 分别为4.6%～10.7%,5.9%～9.9%(n=5)。结论:该方法灵敏、简便、准确,适用于冰片和丹皮酚血药浓度的同时检测和药动学研究。

中药复方液对麻黄素致损伤仔鼠血浆GOT、GPT活性和肝组织Bax蛋白表达的影响

探讨中药复方液对麻黄素致损伤仔鼠血浆谷草转氨酶（GOT）、谷丙转氨酶（GPT）活性和肝组织Bax蛋白表达的影响.选出生5~7d的昆明小鼠,随机分为3组,麻黄素组和中药组采用递增剂量连续腹腔注射0.2mL（2.0,3.0和4.0g·L-1）的麻黄素溶液（1~5d,6~10d,11~15d）,中药组在注射麻黄素1h后分别灌胃0.2 mL（20.0,30.0和40.0g·L-1）的中药复方液,对照组注射和灌胃等量的生理盐水.分别于注射和灌胃药物后第5d,10d,15d时称量仔鼠体重和肝重,用比色法测定仔鼠血浆GOT和GPT活性的变化,用免疫组织化学法检测肝组织Bax蛋白表达的变化.结果表明,麻黄素组仔鼠体重和肝重均低于对照组,仔鼠血浆GOT、GPT活性高于对照组,仔鼠肝组织Bax蛋白阳性表达量高于对照组（P〈0.05或P〈0.01）;中药组仔鼠体重和肝重均高于麻黄素组,仔鼠血浆GOT,GPT活性低于麻黄素组,仔鼠肝组织Bax蛋白阳性表达量低于麻黄素组（P〈0.05或P〈0.01）.表明中药复方液影响麻黄素致损伤仔鼠血浆转氨酶的活性,可降低因麻黄素损伤引起的肝组织Bax蛋白的表达.

抗栓中药对小白鼠体内血栓形成及血浆t-P A 及 P A I的影响

目的：研究复方丹参及川芎嗪注射液中药抗栓胶囊对小白鼠体内血栓形成及血浆组织型纤维溶酶原激活物（ｔ Ｐ Ａ）及其抑制物（ Ｐ Ａ Ｉ）的影响。方法：给不同组小鼠分别注射复方丹参及川芎嗪注射液、以及服用中药抗栓胶囊７ ｄ。用致栓药角叉菜胶诱发小鼠体内血栓形成。发色底物法测定血浆 ｔ Ｐ Ａ 及 Ｐ Ａ Ｉ活性。结果：复方丹参组及中药抗栓胶囊组小鼠尾部平均血栓形成长度百分数比血栓对照组明显缩短，复方丹参组、中药抗栓胶囊组血浆 Ｐ Ａ Ｉ低于血栓对照组，有显著性差异（ Ｐ＜ ００５）。结论：复方丹参、中药抗栓胶囊具有抗栓作用。其作用机理可能主要与可减少血浆 Ｐ Ａ Ｉ的释放有关。

反相高效液相色谱法测定小鼠血浆中β-榄香烯含量

目的：建立小鼠血浆中β-榄香烯的反相高效液相色谱法含量分析方法。方法：小鼠血浆中加入2倍体积的乙腈提取出β-榄香烯用于测定。色谱柱为Hypersil BDS C18（4．6mm×200mm，5μm），流动相为乙腈-水（体积比90：10），检测波长为205nm，流速为1mL／min。结果：血浆中其他组分不影响β-榄香烯的检测，在0．4～24．0mg／L浓度范围内，线性关系良好（Y=160．73X-24．12，r=0．9996），相对回收率〉（95．70±3．07）％。结论：本方法具有简单、快速、准确等特点，适用于测定血浆中β-榄香烯的浓度。

高效液相色谱法测定小鼠血浆中白花丹素及其蛋白结合率

目的:建立小鼠血浆白花丹素浓度的测定方法,测定白花丹素与小鼠血浆的蛋白结合率。方法:用超滤法测定血浆蛋白结合率,用高效液相—紫外检测法测定血浆中药物总浓度及游离药物浓度。结果:白花丹素在0·056～11·2mg/L范围线性良好,标准曲线方程为:y=9910·4x-179·31(r=0·9980)。浓度为0·56、1·12、5·6、11·2mg/L的白花丹素在小鼠血浆的蛋白结合率为(69·4±2·6)%、(76·2±5·3)%、(73·5±4·6)%、(80·4±7·7)%。结论:高效液相色谱法能快速、精密的测定小鼠血浆中白花丹素浓度,白花丹素与小鼠血浆蛋白结合率在70%～80%之间。

去甲基雏菊叶龙胆酮在小鼠体内的药动学研究

目的:研究川东獐牙菜中去甲基雏菊叶龙胆酮在小鼠体内的药动学过程。方法:小鼠一次性腹腔注射去甲基雏菊叶龙胆酮后,用高效液相色谱法检测不同时间间隔血药浓度,计算药动学参数。结果:小鼠一次性腹腔注射0.8g.kg-1的去甲基雏菊叶龙胆酮后其药动学参数为t1/2α为(19.8±4.8)min,t1/2β为(411.7±52.8)min,AUC为(92.2±10.9)mg.min.L-1。结论:本方法灵敏、快速、准确、选择性强,可较好地满足去甲基雏菊叶龙胆酮的药动学研究,为进一步研究药理作用提供依据。

毛细管电泳-电化学发光法测定小鼠血浆中粉防己碱含量

血浆样品用氯仿萃取,所得萃取液在60℃水浴上用吹氮蒸干,用500μL甲醇溶解残渣,所得溶液用电驱动方法进样引入毛细管中进行毛细管电泳（CE）分离。采用的进样电压为12 kV,进样时间为7 s。电泳缓冲溶液为pH 7.5的20 mmol·L^-1磷酸盐缓冲溶液,分离电压为15 kV。电化学发光法（ECL）与CE相联用作为检则方法。ECL反应系在400μL反应池中进行,反应池中盛有5 mmol·L^-1Ru（bpy）32＋溶液和pH 8.0的50 mmol·L^-1磷酸盐缓冲溶液。采用的检测电位为1.15 V（vs.Ag/AgCl）,所测得的ECL强度值与粉防己碱的质量浓度在0.05-80.0 mg·L^-1范围内呈线性关系,其检出限（3S/N）为0.02 mg·L^-1。在小鼠血浆样品的基础上加入粉防己碱标准溶液进行回收试验,测得其回收率在93.3%-95.0%之间。

胺碘酮对小鼠华法林血药浓度的影响

目的探讨胺碘酮对小鼠华法林血药浓度的影响。方法将96只小鼠分为华法林对照组,胺碘酮30、60、120 mg·kg-1组。每天上午给各组小鼠灌华法林,下午灌生理盐水或胺碘酮,7 d后取血用高效液相色谱测定华法林血药浓度。结果与对照组比较,胺碘酮30 mg·kg-1组华法林血药浓升高度差异无统计学意义(P>0.05),胺碘酮60、120 mg·kg-1组华法林血药浓度升高差异有统计学意义(P<0.05);胺碘酮各剂量组间华法林浓度比较差异均有统计学意义(P<0.05)。结论胺碘酮60 mg·kg-1可呈剂量依赖性的抑制华法林的代谢,使后者血药浓度增高。

苜蓿总黄酮对小鼠脂类代谢及氧自由基的影响

试验结果显示:灌胃给予苜蓿总黄酮(TFA)的小鼠全血中甘油三酯(TG)和低密度脂蛋白胆固醇(LDL-C)的含量明显降低(P<0.05),而高密度脂蛋白胆固醇(HDL-C)含量升高,而且其肝脏和肾脏组织匀浆中过氧化物歧化酶(SOD)的活性显著升高(P<0.05),丙二醛(MDA)的含量降低。因此得出结论:TFA具有降血脂、降低低密度脂蛋白胆固醇含量、预防和减轻动脉硬化的作用;同时也具有抑制氧自由基损伤、防止脂质过氧化的作用。

冷应激对老年小鼠体温、血糖浓度和血浆MDA水平的影响

本文研究了遭受冷应激的老年小鼠的体温、血糖浓度及血浆MDA的水平。结果表明,与对照组比较应激使老年小鼠体温发生显著下降,血糖上升不足及血浆MDA水平显著升高。提示老年小鼠应激承受力减弱,导致体温的稳定性下降,这可能与血糖上升不足有关而脂质过氧化反应增强可能会进一步削弱应激引起的升血糖反应。

硒茶与非硒茶对高胆固醇饮食小鼠血脂的影响

目的 :比较硒茶与非硒茶对脂质水平异常的动物个体血脂及脂蛋白代谢的影响。方法 :将高胆固醇饮食小鼠分为三组 ,分别常规饮水、饮用硒茶及非硒茶 ,14周后测定血浆TC、TG、LDL -C及HDL -C水平。结果 :高脂膳食 +非硒茶组TC、TG显著低于高脂膳食组 (P <0 .0 1) ,LDL -C及HDL -C与高脂膳食组无差异 ;而高脂膳食 +硒茶组TC、TG及LDL -C均显著低于高脂膳食组 (P <0 .0 1,P <0 .0 5 ) ,HDL -C高于高脂膳食组及高脂膳食 +非硒茶组 (P <0 .0 5 )。结论 :长期饮用硒茶能显著改善高胆固醇饮食小鼠血脂及脂蛋白代谢状况 ,且效果明显优于非硒茶。