胶束化色胺酮的制备及表征

目的通过制备胶束化色胺酮,增加色胺酮的溶解度,并进一步提高其生物利用度。方法:以酸敏感的腙键连接聚乙二醇和色胺酮,并通过透析法,将聚乙二醇化色胺酮进一步制备成胶束。用动态光散射法测定胶束的粒径分布用透射电镜观察胶束的形貌。通过芘荧光探针法测定胶束的临界胶束浓度。测定胶束在不同pH下的药物释放情况(pH5.5和7.4)。采用薄层色谱法和高效液相色谱法研究腙键的断裂行为。通过CCK-8法比较生理pH和酸性pH下,色胺酮和聚乙二醇化色胺酮胶束(PTMs)对MCF-7细胞的体外细胞毒性。结果:与色氨酸相比,PTMs的溶解度提高了1493倍。制备的胶束粒径为228.8 nm,PDI为0.1,形貌为球形。PTMs的临界胶束浓度为3.5×10^-7mol/L,较低的CMC值表明制备的胶束稳定性高,便于进一步使用。腙键可在酸性条件下发生断裂,且在pH 5.5下,12 h内95%的色胺酮从胶束中释放,而在生理pH下(pH 7.4),药物释放缓慢。在生理条件下胶束的细胞毒性低于色胺酮,说明胶束化色胺酮可降低药物毒性及胶束在生理条件下有一定的稳定性。而在pH 5.5时,色胺酮胶束与色胺酮的细胞毒性相近表明胶束可响应肿瘤细胞内的低pH值成功实现药物释放。结论:胶束化色胺酮不仅能有效改善色胺酮的溶解度,有利于进一步提高其生物利用度,而且是一种很有应用前景的肿瘤靶向前药。     

青年人进驻不同海拔高度时血浆NO和NOS的变化

一氧化氮(ＮＯ)是在自然界中发现的10个最小分子中的1个,并在中枢神经、周围神经和心血管系统中充当化学信使的重要角色。ＮＯ和一氧化氮合酶(ＮＯＳ)参与高原缺氧引起机体的生理病理性变化已受到人们的重视,研究高原居住人群ＮＯ和ＮＯＳ的变化对高原疾病防治和提高高原人群的整体健康水平具有重要意义。1　方法1998年4月我们对进驻海拔3700ｍ(10人)、5070ｍ(10人)和5380ｍ(10人)不同海拔高度8～10ｄ的健康士兵检测了血浆ＮＯ和ＮＯＳ的含量,并以平原(1400ｍ)10名健康士兵为对照。(1)检测对象　汉族男性,平原出生,年龄18～24岁,平均19.8岁,…     

川芎当归不同配伍对偏头痛大鼠体内5-HT, NO和NOS的影响研究

目的:研究不同比例的川芎当归配伍对偏头痛大鼠体内内源性物质影响。方法:采用雄性SD大鼠,皮下注射硝酸甘油混悬液制作偏头痛大鼠模型,以偏头痛模型大鼠体内三种内源性物质5-HT、NO和NOS含量为考察指标,观察给予川芎-当归不同配比(1:9,2:8~9:1)提取液后,模型大鼠体内5-HT、NO和NOS含量变化。结果:在不同配伍比例川芎当归给药后,模型大鼠脑内5-HT含量随川芎比例增加而增加,当川芎比例超过5:5是变化显著,达到8:2时变化最显著;NO而NOS含量变化差异不显著。结论:川芎当归药预防给药对能够减少脑内5-HT在偏头痛发作时的消耗,发挥收缩血管作用,且随着川芎比例增大,作用逐渐增强;阿魏酸对偏头痛的预防有效,但不是唯一有效成分,不能作为药对作用的决定因素。     

色胺酮对乳腺癌MCF-7细胞凋亡的诱导作用

目的:探讨色胺酮(Tryptanthrin,Try)对人乳腺癌MCF-7细胞增殖和凋亡的影响。方法:利用MTT方法检测Try(1.56-100μmol/L)对细胞增殖的影响;透射电镜观察细胞的形态学改变;流式细胞术检测细胞周期、凋亡情况及线粒体跨膜电位等指标。结果:MTT结果显示,Try在12.5-100μmol/L浓度范围内能明显抑制MCF-7细胞的增殖,并具有时间和浓度依赖性;透射电镜下可见Try作用48h后,MCF-7细胞有典型的凋亡样改变。Annexin V-FITC与PI双染,流式细胞仪检测结果显示:50、100μmol/LTry作用后,细胞的凋亡情况明显,与对照组相比差异显著;且影响了MCF-7的细胞周期分布,将细胞阻滞于G1期,抑制其DNA的合成;并导致细胞线粒体跨膜电位下降。结论:色胺酮能明显抑制MCF-7细胞增殖并具有诱导细胞发生凋亡的作用。     

色胺酮对心肌缺血大鼠心功能和心肌组织COX-2蛋白表达等影响

目的:观察色胺酮对结扎冠状动脉所致心肌缺血大鼠心功能、心肌病理和心肌中COX-2的影响.方法采用结扎冠状动脉左前降支致心肌缺血模型,色胺酮0.10、0.20、0.30g/kg灌胃给药21天后,观察色胺酮对心肌缺血大鼠心功能、心肌病理和心肌组织COX-2蛋白的影响.结果①色胺酮0.10g/kg对心肌缺血大鼠的心脏功能有明显的改善作用,能增强心肌收缩力,改善心肌舒张的顺应性,表现为左心室内收缩压、左心室内压最大变化速率±dp/dtmax明显升高,与模型组比较有明显差异(P<0.05);②0.10g/kg色胺酮对结扎冠状动脉所致心肌缺血大鼠能明显减轻心肌变性,心肌纤维增生和淋巴细胞浸润;③色胺酮明显抑制缺血心肌组织中COX-2蛋白,其抑制作用随剂量的增大而逐渐增强.结论0.10g/kg色胺酮对大鼠结扎冠状动脉所致心肌缺血损伤具有明显的保护作用.     

基于高通量测序技术研究色胺酮对溃疡性结肠炎小鼠肠道菌群的影响

目的 研究色胺酮对葡聚糖硫酸钠(DSS)诱导溃疡性结肠炎(UC)模型小鼠肠道菌群的影响。 方法 用3% DSS自由饮用7 d诱导昆明种小鼠UC模型,灌胃125 mg/kg柳氮磺胺吡啶作为阳性对照,色胺酮组以78 mg/kg水溶液灌胃给药,观察小鼠给药前后的体征,进行疾病活动指数(DAI)评分,观察结肠组织形态变化;ELISA法检测小鼠血清中IL 6、IL 8、IL 10、IL 1β和TNF α水平变化;每组收集5个粪便样本,利用Illumina MiSeq测序平台采用细菌的16S rRNA进行V4-V5区高通量测序,对测序结果进行生物信息学分析。 结果 色胺酮和阳性药干预后,DAI评分表明小鼠体征有所改善,HE染色显示UC小鼠结肠组织损伤有一定程度修复,炎性细胞减少;ELISA检测显示小鼠血清中促炎性细胞因子IL 6、IL 8、IL 1β和TNF α水平下调,抑炎性细胞因子IL 10水平上升;测序后菌群物种注释及生物多样性分析显示色胺酮干预后UC小鼠失调的厚壁菌门/拟杆菌门比例得到一定程度恢复,菌群多样性有一定程度回调,菌群结构向正常状态恢复;肠道菌群LEfSe组间差异显著性分析结果表明4个菌属Erysipelotrichaceae_UCG_003、Sellimonas、Rikenellaceae_RC9_gut_group和Tyzzerella丰度可能与色胺酮对UC治疗作用相关。 结论 色胺酮治疗UC小鼠后,小鼠体征、结肠组织病理结构、细胞因子水平及肠道菌群变化等结果均表明色胺酮对UC小鼠肠道微生态具有调节作用,为色胺酮抗UC作用机制研究提供了新思路。     

血立停胶囊对早孕大鼠RU486药流后NO、NOS表达的影响

目的:研究血立停胶囊对早孕大鼠RU486药物流产后的子宫平滑肌一氧化氮(NO)及一氧化氮合酶(NOS)水平变化的影响。方法:选择妊娠Wistar大鼠,随机分为5组,即对照组,米非司酮组,大剂量血立停组,小剂量血立停组,催产素组。于妊娠第7天,开始相应处理,妊娠第14天分别监测早孕大鼠RU486药物流产后的子宫平滑肌一氧化氮(NO)及一氧化氮合酶(NOS)水平后处死。结果:大剂量血立停可明显降低大鼠子宫肌组织匀浆中NO、NOS含量,与对照组比较差异有显著性(P<0.05)。结论:血立停胶囊可降低子宫肌组织匀浆中NO、NOS水平,从而起到对药物流产后阴道出血的治疗作用。     

豚鼠肺内NOS之分布及实验性哮喘对其活性的影响

用组织化学方法对豚鼠肺内一氧化氮合酶（NOS）进行定位研究．并观察正常对照组和支气管哮喘组豚鼠肺内NOS分布及活性变化。结果显示;（1）正常豚鼠各级支气管、肺泡管、肺泡囊上皮细胞均成NOS阳性反应,气道上皮下平滑肌细胞呈NOS阴性反应。（2）哮喘豚鼠各级支气管、肺泡管、肺泡囊上皮细胞呈NOS强阳性反应,气道上皮下平滑肌细胞里NOS阳性反应。（3）两组肺泡上皮细胞均呈NOS阴性反应。（4）两组肺内动,静脉血管内膜均呈NOS阳性反应。结果提示一氧化氮不仅对肺具有一定的生理作用,而且可能参与哮喘的病理生理过程。     

小鼠血浆TNF—α、NO、NOS水平与脑不对策的关系

研究脑不对称对单核巨噬细胞 (Mo/MФ)的影响。选用雌性 4周龄Balb/c健康小鼠 ,用伸爪取食法根据右利分将小鼠区分为左利、右利和双利三组。细菌脂多糖 (lipopolysaccharide ,LPS)腹腔注射两小时后取血浆检测肿瘤坏死因子 α(TNF α)、一氧化氮 (NO)、一氧化氮合酶 (NOS)水平。结果显示 :( 1)各组间血浆TNF α和NO、NOS水平的变化 :正常生理组NO水平为双利组高于左利组 (P <0 0 5 ) ;LPS刺激后双利组和右利组血浆TNF α水平高于左利组 (P <0 0 5 ) ,NO水平为双利组高于右利组 (P <0 0 1)和左利组 (P <0 0 5 ) ,NOS水平在三组小鼠间无明显差别。 ( 2 )血浆TNF α、NO、NOS与右利分有一定的相关关系 ,各项指标随右利分的变化趋势多为双利>右利和左利 ,形成一以双利分段 (右利分为 2 1～ 2 9)为高峰的弓形向上的曲线 ,随右利分的增加或减少曲线向两侧延伸和下降。上述结果提示 ,脑不对称小鼠单核 /巨噬细胞功能存在差异。脑不对称对免疫功能的影响程度与右利分有关 ,各免疫指标随右利分的变化趋势多为一以双利分段为高峰的弓形向上的曲线     

降气化痰推拿法对慢性哮喘幼鼠气道重塑及血清NO水平的影响

摘要 目的：探究降气化痰推拿法对慢性哮喘幼鼠的治疗效果,为优化临床中小儿慢性哮喘治疗方案积累理论基础。方法：选取40只SPF级3周SD雌性小鼠,随机数字表法为对照组、模型组、药物组和推拿组,每组各10只。模型组、药物组和推拿制备慢性哮喘模型后,药物组使用地塞米松治疗,对照组注射等量生理盐水,推拿组在药物组基础上加入降气化痰推拿手法治疗,模型组不进行干预。比较各组幼鼠肺组织HE染色结果、支气管肺泡灌洗液（BALF）中炎性细胞水平和Th17/Treg细胞比例,并检测肺组织中Treg与Th17相关基因表达量。结果：与模型组比较,药物组和推拿组的支气管周围组织的炎性浸润较轻,肺泡壁及支气管壁的结构较为完整,气道可见少量杯状细胞增生,气道壁总厚度和气道平滑肌厚度增加较少,且推拿组大鼠的组织炎性浸润程度、气道壁总厚度和气道平滑肌厚度均低于药物组（P<0.05）;炎性细胞水平方面,对照组幼鼠的BALF细胞总数较低,白细胞分类中单核细胞数最多。模型组幼鼠的BALF中细胞总数明显增高,且白细胞分类中以中性粒细胞、嗜酸性粒细胞为主,白细胞总数和嗜酸性细胞总数明显高于对照组（P<0.05）;与模型组相比,药物组和推拿组的细胞总数及中性粒细胞和嗜酸性粒细胞的比例明显较低,但仍明显高于对照组,且药物治疗组高于推拿组（P<0.05）;与对照组相比,其余各组幼鼠的BALF中Th17比例明显增高,Treg比例降低,Th17/Treg比值增高（P<0.05）;与模型组相比,药物组和推拿组的Th17比例、Th17/Treg降低,且推拿组低于药物组,Treg比例升高,且推拿组高于药物组（P<0.05）;与对照组对比,其余各组幼鼠的IL-6、IL-10、IL-23、TGF-β mRNA表达水平均升高,与模型组相比,药物组与推拿组幼鼠表达水平均有所降低,且推拿组低于药物组（P<0.05）;与对照组相比,其余各组幼鼠的NO、IgE水平升高,与模型组相比,药物组和推拿组的NO、IgE水平降低,且推拿组低于药物组（P＜0.05）。结论：降气化痰推拿法可通过调节Th17/Treg比例及相关炎性因子基因表达和白细胞类型,调控血清NO、IgE水平和炎性反应水平,进而改善气道重塑,发挥治疗作用。     

精氨酸甲基转移酶1和核因子кB在哮喘豚鼠中的表达变化

目的:探讨共激活因子相关的精氨酸甲基转移酶1和核因子-кB在豚鼠支气管哮喘模型气道和肺组织的表达变化.方法:24只白色雄性豚鼠随机分为:①正常对照组;②哮喘组.卵清蛋白致敏并激发后采用间接免疫荧光法检测气道及肺组织精氨酸甲基转移酶1和核因子NF-кB(P65)的表达水平,探讨其在哮喘中可能的作用机制.结果:CARM1和NF-кB(P65)在对照组和哮喘组均有阳性表达,主要在支气管-终末细支气管上皮细胞和肺组织成纤维细胞胞核表达.正常对照组和哮喘组CARM1和NF-кB(P65)的表达差异均有统计学意义.结论:在哮喘豚鼠气道上皮及肺组织CARM1和NF-кB(P65)在细胞胞核高表达,提示CARM1可能通过增强募集NF-кB到相关位点激活NF-кB信号转导通路,并启动了多种前炎性基因和免疫调节基因的转录激活从而诱发哮喘炎症反应.     

硬枝碱蓬多糖对免疫抑制小鼠血清中NO含量及NOS活性的影响

采用氢化可的松皮下注射小鼠建立免疫抑制模型,观察不同浓度硬枝碱蓬多糖(100、200和400mg/kg·d)灌服小鼠后(模型对照组灌服等体积生理盐水),对免疫抑制小鼠血清中NO含量及TNOS、iNOS活性的影响。结果表明:3个试验组免疫抑制小鼠血清中NO含量及TNOS、iNOS活性均明显降低。其中,硬枝碱蓬多糖低、中浓度组iNOS活性显著低于模型对照组(P<0.05),高浓度组iNOS活性极显著低于模型对照组(P<0.01);硬枝碱蓬多糖低、中、高浓度组NO含量及TNOS活性与模型对照组相比差异均极显著(P<0.01),且呈现明显的浓度依赖效应。结果提示,硬枝碱蓬多糖可通过抑制NOS并最终抑制NO的细胞毒性作用而对正常组织细胞发挥保护作用,并进一步增强小鼠的免疫力。     

Mechanism of Tuberculostasis in Mammalian Serum II. Induction of Serum Tuberculostasis in Guinea Pigs

The growth of tubercle bacilli in serum samples of untreated animals depends upon the availability of ionic iron which serves as a growth factor in supporting bacillary multiplication. The amount of available iron in serum is determined by the ratio between iron-saturated and iron-free transferrin; a low value for the ratio is associated with tuberculostasis (e.g., human serum, 0.4), whereas a high value is associated with the growth-supporting quality (e.g., guinea pig serum, 5.6). The treatment of guinea pigs with lipopolysaccharide of Escherichia coli or tuberculous cell wall material consistently and significantly reduced serum iron levels; a similar but less striking effect was observed in BCG-vaccinated animals. Pronounced differences were observed in the time of appearance and duration of serum hypoferremia; in lipopolysaccharide-treated animals, it appeared in 1 day and lasted for several days, whereas in BCG-vaccinated animals it appeared in about 2 weeks and lasted for much longer time periods. The induced hypoferremia was always associated with the concomitant development of serum tuberculostasis which could be neutralized by the addition of iron. These results indicate, therefore, that the mechanism of induced serum tuberculostasis in lipopolysaccharide- or tuberculous cell wall-treated and BCG-vaccinated guinea pigs is the same as that present in tuberculostatic sera of untreated animals.     

Temperature Effects on Legionella pneumophila Killing by and Multiplication in Phagocytes of Guinea Pigs

We examined the effects of temperature on the interaction between Legionella pneumophila and phagocytes of guinea pigs. The body temperatures of guinea pigs infected with a sublethal dose (1.2 × 10⁴ CFU) or a lethal dose (1.0 × 10⁵ CFU) of L. pneumophila elevated from 38.4±0.15 C to 40.2±0.42 C or 40.3 ± 0.62 C, respectively. The intracellular bacterial killing by and bacterial proliferation in the phagocytes were examined at 33, 37, 40, and 42 C, using in vitro culture systems of peritoneal macrophages or polymorphonuclear leukocytes (PMN) of guinea pigs. In all the macrophages incubated at different temperatures, significant intracellular bacterial killings were observed at 4 hr after in vitro phagocytosis. After 24 hr of incubation, there was about a 100-fold increase of CFU and the number reached a maximum after 48 hr of incubation in the macrophages incubated at 42 C as well as 37 and 40 C, suggesting that macrophages support the intracellular bacterial growth in hyperthermia. In the PMN, L. pneumophila CFU 4 hr or 12 hr after the infection were significantly lower at 42 C than those at 37 C (P<0.05), indicating that the bactericidal capacity of PMN was enhanced at 42 C compared to 37 C. However, in all the PMN incubated at different temperatures, there were about 10-fold increases of CFU 24 hr after the infection, suggesting that PMN as well as macrophages support intracellular bacterial growth in hyperthermia. The extracellular bacterial growth was examined at 33, 37, 40, and 42 C in buffered yeast extract (BYE) broth or RPMI 1640 medium containing 50% guinea pig serum as a permissive or non-permissive liquid medium for the bacterial growth, respectively. Inhibition of bacterial growth in BYE broth at 42 C, and a decrease of CFU in RPMI 1640 medium containing 50% guinea pig serum at 42 C were observed. In conclusion, hyperthermia may be beneficial by restricting extracellular bacterial survival, but it exerts no beneficial effect on the restriction of intracellular bacterial growth in phagocytes, though PMN showed enhanced initial killing at 42 C. These results suggest that fever, or hyperthermia itself, may not largely contribute as a nonspecific host defense early in the course of legionellosis.     

变压器噪声暴露对豚鼠应激、肝肾及免疫功能的影响

目的:探讨短时间内变压器噪声暴露对豚鼠应激、肝肾及免疫功能的影响。方法:取32只健康成年(5-6月龄)豚鼠随机分为实验组和对照组,每组16只。实验组给予录制的变压器噪声(声压级范围40.8-55 d B SPL,频谱范围150-2000 Hz)连续暴露28天,10小时/天(晚10点到早上8点),对照组在相同条件下饲养,无噪声暴露。噪声暴露结束后,对实验豚鼠的应激、肝肾及免疫功能进行定量评估比较。结果:噪声暴露28天后实验组豚鼠的应激状态指标(ACTH、血清皮质醇)与对照组比较差异无统计学意义(P0.05),主要肝肾功能指标与对照组比较差异无统计学意义(P0.05),免疫相关指标(Ig G、Ig A、Ig E、IL-1、IL-2)比较差异无统计学意义(P0.05)。结论:声压级范围为40.8-55 d B SPL、频谱范围为150~2000 Hz的变压器噪声连续暴露28天(10小时/天)对成年豚鼠应激、肝肾及免疫功能无明显影响。     

Effects of Animal-Assisted Activities with Guinea Pigs in the Primary School Classroom

ABSTRACT

This study investigated the effects of a classroom-based animal-assisted activities (AAA) program with guinea pigs on the social functioning of primary school children. We hypothesized that participants in the experimental condition (n = 64), compared with a waitlist control group (n = 64), would demonstrate improvements in social functioning following the program. Parents and teachers used the Social Skills Rating System (SSRS) to evaluate the social skills and problem behaviors of 128 participating children (age range = 4.8 to 12.7 years) before and after an 8-week period. Teachers also rated academic competence at both time points. Children who participated in the AAA program demonstrated significantly greater improvements in social functioning than their control group peers, as defined by greater increases in social skills (teacher SSRS) and decreases in problem behaviors (parent and teacher SSRS). There were no significant differences between the groups in academic competence. AAA participants demonstrated significant increases in social skills and decreases in problem behaviors from pre- to post-program on the teacher version of the SSRS. Control group participants did not show significant changes on these measures. These findings suggest that an AAA program with guinea pigs may be a feasible addition to the primary school classroom in order to improve social functioning. Further component analysis will be necessary to determine whether the animal is the active ingredient in AAA programs of this nature.     

Different Effects of Propofol and Isoflurane on Cochlear Blood Flow and Hearing Function in Guinea Pigs

Objectives

To investigate the effects of isoflurane and propofol on mean arterial pressure (MAP), cochlear blood flow (CoBF), distortion-product otoacoustic emission (DPOAE), and the ultrastructure of outer hair cells (OHCs) in guinea pig cochleae.

Methods

Forty-eight male guinea pigs were randomly assigned to one of six treatment groups. Groups 1 to 3 were infused (i.v.) with a loading dose of propofol (5 mg/kg) for 5 min and three maintenance doses (10, 20, or 40 mg kg⁻¹·h⁻¹, respectively) for 115 min. Groups 4 to 6 were inhaled with isoflurane at concentrations of 1.15 vol%, 2.30 vol% or 3.45 vol% respectively for 120 min. CoBF and MAP were recorded prior to and at 5 min intervals during drug administration. DPOAE was measured before, immediately after, and 1 h after administration. Following the final DPOAE test, cochleae were examined using scanning electron microscopy.

Results

Propofol treatment reduced MAP in a dose-dependent manner. CoBF and DPOAE showed increases at propofol maintenance doses of 10 and 20 mg kg⁻¹·h⁻¹. Inhalation of isoflurane at concentrations of 2.30 vol% and 3.45 vol% reduced MAP and CoBF. DPOAE amplitude increased following inhalation of 1.15 vol% isoflurane, but decreased following inhalations of 2.30 vol% and 3.45 vol%. Cochlear structure was changed following inhalation of either 2.30 vol% or 3.45 vol% isoflurane.

Conclusions

Propofol could decrease MAP and increase both CoBF and DPOAE without affecting OHC structure. Inhalation of isoflurane at concentrations >2.30 vol% decreased CoBF and DPOAE, and produced injury to OHCs.