峨眉杠柳的化学成分研究

目的研究峨眉杠柳的化学成分。方法利用色谱技术进行分离纯化,以波谱技术进行结构鉴定。结果从峨眉杠柳中分离得到7个化舍物,鉴定为23-羟基齐墩果酸（1）、β谷甾醇（2）、27-羟基-α-香树脂醇（3）、熊果酸（4）、（＋）-1-羟基松脂酚（5）、胡萝卜苷（6）、杠柳苷（7）。结论该植物有较好的研究价值。     

杠柳苷元抑制人乳腺癌细胞增殖作用的研究

目的 观察杠柳苷元(periplogenin)对人乳腺癌细胞MCF7的抑制作用并考察其抑制作用的机理.方法 使用四氮唑盐(MTT)法测定杠柳苷元对MCF7细胞的增殖抑制效果,并使用蛋白免疫印迹法测定其中凋亡蛋白的表达.结果 杠柳苷元对MCF7细胞起到了明显的抑制作用,其抑制作用呈剂量依赖性,蛋白免疫印迹法也观察到杠柳苷元促进MCF7细胞内凋亡相关蛋白的表达,其表达呈时间依赖性.结论 杠柳苷元具有显著的体外抑制MCF7细胞增殖的作用,其抑制作用是通过促进MCF7细胞凋亡而发挥的.     

杠柳毒苷在大鼠体肠吸收动力学研究

目的考察杠柳毒苷在大鼠小肠内的吸收情况。方法采用大鼠在体单向肠灌流模型,用高效液相色谱法对灌流液中的杠柳毒苷进行分析。结果杠柳毒苷在不同浓度下未发现饱和现象,其吸收速率常数(Ka)和表观吸收系数(Papp)基本保持不变。结论杠柳毒苷在小肠的吸收机制为被动扩散,在临床应用及制剂研究中应考虑其吸收机制。     

基于斑马鱼模型的杠柳毒苷肝毒性评价

目的 基于斑马鱼模型初步探讨杠柳毒苷的毒性靶器官和潜在作用机制。方法 将斑马鱼(4 days post fertilization,4 dpf)暴露于不同浓度(低浓度1μg·mL^-1,高浓度1.5μg·mL^-1)的杠柳毒苷溶液中24 h,统计杠柳毒苷对斑马鱼的致死率;10%致死剂量下(10)暴露24 h后,基于肝脏面积变化,吖啶橙染色,肝脏组织病理切片,谷丙转氨酶(ALT)与谷草转氨酶(AST)活性变化等指标评价杠柳毒苷的肝毒性;通过网络药理学与分子对接技术预测杠柳毒苷肝毒性潜在作用机制,并通过实时荧光定量PCR(real-time PCR,RT-PCR)对预测靶点进行验证。结果 杠柳毒苷对斑马鱼的亚致死浓度(LC₁₀)为1.612 6μg·mL^-1,亚致死浓度暴露条件下,与空白组相比,杠柳毒苷能够引起斑马鱼肝脏面积减小和明显的肝脏细胞凋亡,组织病理切片结果显示杠柳毒苷能够诱导肝细胞出现结构排列松散紊乱以及明显空泡化等病理特征。此外,杠柳毒苷能够显著升高斑马鱼ALT...     

杠柳毒苷在大鼠体内的组织分布

目的 考察杠柳毒苷在大鼠体内的组织分布.方法 采用高效液相色谱法测定大鼠静脉注射杠柳毒苷后,组织样品中杠柳毒苷的质量浓度.结果 大鼠iv杠柳毒苷0.74 mg/kg,2、10、20 min后肝中杠柳毒苷的质量浓度分别为(2.447±0.687)、(0.6310±0.272)、(0.2924±0.228)μg/g,在肝中...     

杠柳毒苷在大鼠体内的药动学研究

目的考察杠柳毒苷iv给药在大鼠体内的药动学过程。方法大鼠iv给予低、中、高剂量(0.37、0.74、1.48mg/kg)杠柳毒苷后于不同时间点采血,血浆样品经甲醇沉淀蛋白-C18固相萃取处理,HPLC-UV法测定不同时间点大鼠血浆中杠柳毒苷药物浓度。用DAS药动学数据软件计算药动学参数。结果低、中、高剂量组杠柳毒苷的分布相半衰期t1/2α分别为1.49、2.32、3.48min,消除相半衰期t1/2β分别为14.00、12.37、15.44min,无显著性差异。AUC0-60分别为8.581、19.782、50.615mg/L·min,与剂量呈线性相关。结论杠柳毒苷iv给药在大鼠体内分布及消除迅速,其体内过程符合二房室模型,在0.37～1.48mg/kg符合线性动力学过程。     

杠柳苷元对肥大细胞脱颗粒及释放组胺影响的研究

目的:研究香加皮提取单体化合物杠柳苷元对大鼠和小鼠肥大细胞脱颗粒及组胺释放的影响。方法:大鼠腹腔注射百日咳疫苗、后腿注射卵白蛋白致敏,用于测定肥大细胞脱颗粒反应及制备抗血清;取致敏大鼠的血清稀释后对小鼠进行腹腔注射,测定肥大细胞脱颗粒反应;以荧光分光光度法测定组胺浓度。结果:杠柳苷元对体外培养肥大细胞的组胺释放有显著的抑制作用,实验剂量即可使组胺释放浓度降低(69.4±8.6)%,其抑制作用呈显著的剂量依赖关系;杠柳苷元对抗原致敏大鼠肥大细胞的组胺释放也有显著的抑制作用,在20μg·mL-1浓度时即可使组胺释放浓度减少73.55%;杠柳苷元口服给予致敏小鼠后,在50mg·kg-1剂量时即可使小鼠组胺释放浓度减少80%以上,并呈显著的剂量依赖关系。结论:杠柳苷元对体外培养的肥大细胞、致敏大鼠肥大细胞的组胺释放有显著的抑制作用;口服给予杠柳苷元可使小鼠显著减少肥大细胞的组胺释放。鉴于肥大细胞脱颗粒及组胺释放在炎症反应中的作用,可认为杠柳苷元是香加皮具有抗炎作用的有效成分之一。     

杠柳毒苷体外抑制肝癌细胞和乳腺癌细胞增殖的实验研究

目的探讨杠柳毒苷在体外对人乳腺癌MDA—MB．468细胞和人肝癌HepG2细胞增殖的影响。方法MTT法观察杠柳毒苷对人乳腺癌MDA-MB-468细胞和人肝癌HepG2细胞增殖的抑制作用,流式细胞术观察杠柳毒苷对两种肿瘤细胞的细胞增殖周期作用。结果与对照组比较,杠柳毒苷能明显抑制两种肿瘤细胞的增殖,其抑制率与药物浓度和作用时间呈正相关。流式细胞仪检测发现,杠柳毒苷对乳腺癌MDA-MB-468细胞和肝癌HepG2细胞持续作用24h后,可以使GdG1期细胞增多,G2／M期细胞减少。结论杠柳毒苷具有抑制乳腺癌MDA-MB-468细胞和肝癌HepG2细胞增殖的作用,并可将乳腺癌MDA-MB-468细胞和肝癌HepG2细胞的细胞生长周期阻滞在G0／G1期。     

Chemical constituents from the roots of Periploca sepium with insecticidal activity

Five compounds were isolated from the root powder of Periploca sepium. By mainly HR-ESI-MS, ¹H, ¹³C, and 2D NMR spectral data, they were characterized as periplocoside X (1), oligasaccharide A (2), periplocoside A (3), periplocoside E (4), and periplocoside N (5), respectively. Compounds 1–5 were found to possess insecticidal activities against the red imported fire ant. Among the compounds, periplocoside X showed significant activity with LD₅₀ values of 748.99, 116.62, 2169.58, and 3079.33mg/l against soldiers, workers, males, and alate females of red imported fire ant, respectively.     

Fluorescence Localization and Comparative Ultrastructural Study of Periplocoside NW from Periploca sepium Bunge in the Midgut of the Oriental Amyworm,Mythimna separata Walker (Lepidoptera: Noctuidae)

Periplocoside NW (PSNW) is a novel insecticidal compound isolated from the root bark of Periploca sepium Bunge and has potent stomach toxicity against some insect pests. Previous studies showed that the Mythimna separata larva is sensitive to PSNW, but the Agrotis ispilon larva is insensitive. In this study, preliminary target localization on the midgut of M. separata larvae was conducted via a fluorescence labeling technique. A comparative ultrastructural study on the effects of PSNW on the midguts of M. separata and A. ispilon larvae was performed. Symptom observation results showed that typical stomach toxicity was induced by PSNW in M. separata larvae. Fluorescence localization results showed that PSNW binds to the midgut cells of M. separata larvae. Ultrastructure observations showed destruction of the microvilli, organelle, and cytomembrane in the midgut cells of M. separata larvae, whereas no obvious changes were observed in midgut cells of A. ispilon larvae. These results were consistent with the insecticidal activity of PSNW. Therefore, PSNW might act on the midgut tissues of the insects, and one or more binding sites of PSNW may exist in M. separata larvae midgut cell cytomembranes.     

低氧培养环境对脐带间充质干细胞增殖及生物学特性的影响

目的 研究低氧环境对人脐带间充质干细胞（hUCMSCs）增殖及生物学特性的影响。方法 分别在5% O₂和21% O₂环境下培养hUCMSCs,使用群体倍增时间（PDT）评价hUCMSCs的增殖情况;流式细胞术检测细胞表面标志CD73、CD90、CD105、CD34、CD45、HLA-DR的表达;培养基诱导分化后,茜素红-S对含钙骨细胞染色检测成骨诱导分化,油红O对脂滴染色检测成脂诱导分化,阿尔新蓝8GX对蛋白多糖检测软骨诱导分化;羧基荧光素二醋酸盐琥珀酰亚胺酯（CFSE）染色法检测hUCMSCs对植物血凝素P（PHA-P）刺激的外周血单个核细胞（peripheral blood mononuclear cell,PBMC）的增殖抑制作用,并应用流式细胞术检测对CD8+T细胞的抑制作用;实时荧光定量PCR（qRT-PCR）法检测低氧诱导因子1α（HIF-1α）、血管内皮生长因子（VEGF）、肝细胞生长因子（HGF）、胰岛素样生长因子2（IGF-2）、转化生长因子β（TGF-β）、碱性成纤维细胞生长因子（bFGF）、基质细胞衍生生长因子1（SDF-1）、神经生长因子（NGF）mRNA表达;试剂盒法检测细胞培养上清中IGF-2浓度。结果 21% O₂和5% O₂环境培养的hUCMSCs表面标志CD73、CD90、CD105的表达均为阳性（＞95%）,CD34、CD45、HLA-DR的表达均为阴性（＜2%）,均具有成骨、成脂、成软骨三系分化的能力;5% O₂组的PDT均显著小于21% O₂组（P＜0.05）;PBMC经PHA-P刺激后观察到细胞增殖聚集,与hUCMSCs共培养后几乎没有聚集出现,与PBMC＋PHA-P组比较,PBMC＋PHA-P＋hUCMSCs（21%或5% O₂）组子代细胞明显减少,PBMC＋PHA-P＋hUCMSCs（5% O₂）组PBMC抑制率为（61.44±0.92）%,与PBMC＋PHA-P＋hUCMSCs （21% O₂）抑制率（60.48±4.00）%相当,无统计学差异,且两组hUCMSCs对CD8+T细胞的抑制作用无统计学差异。与21% O₂组比较,5% O₂组hUCMSCs HIF-1α、IGF-2、SDF-1、HGF、VEGF、bFGF、NGF mRNA表达水平显著升高（P＜0.05、0.001）,TGF-β无明显变化; 5% O₂组hUCMSCs培养上清IGF-2水平显著高于21% O₂组（P＜0.001）。结论 5% O₂环境可使hUCMSCs的增殖能力和相关生长因子的表达增强,尤其是IGF-2,但依然保持着与21% O₂环境培养的hUCMSCs相似的表型、分化能力以及淋巴细胞增殖抑制能力。     

不同发育阶段儿童癫痫的用药特点及不良反应研究进展

癫痫是儿童神经系统最常见的疾病之一,目前药物治疗仍是首选方案。儿童癫痫药物治疗与成人有所不同,对不同发育阶段的儿童的癫痫治疗需特殊考虑。婴幼儿期:脏器发育不完善,抗癫痫药物的选择应考虑其安全性;学龄前期和学龄期:学习、认知和社会心理发展的关键时期,抗癫痫药物选择应充分考虑其对认知功能的影响;青春前期和青春期:抗癫痫药物治疗要考虑其长期用药对患儿生长发育各个环节的影响。因此,本文综述了其药物治疗进展及搜集了相关的循证医学证据。     

Development and Validation of Two Solid-Phase Enzyme Immunoassays (ELISA) for Quantitation of Human Epidermal Growth Factors (hEGFs)

Purpose. The purpose of the present investigation was to develop and validate two separate enzyme-linked immunosorbent assays (ELISA) for quantitation of exogenous human epidermal growth factor (hEGFl -53) and its truncated fragment (hEGFl-48) in rat plasma. Methods. The present assay systems were based on the sandwiching of the antigen between a monoclonal mouse anti-hEGFl-53 antibody, pre-coated on a 96-well polystyrene plate, and a polyclonal rabbit anti-hEGFl-48 antibody, which is then detected with a peroxidase-labeled goat anti-rabbit antibody. Results. The calibration curves for hEGFl-48 and hEGFl -53 in plasma were validated over a concentration range of 7.8–250 and 62.5–1000 pg/ml, respectively. Determined from replicate assays of hEGFl-48 quality control samples, the intra-assay precision and accuracy were 8.8% RSD and within ± 9.8%; and the inter-assay precision and accuracy were 14.8% RSD and within ± 9.7% RE, respectively. Determined from replicate assays of hEGFl-53 quality control samples, the intra-assay precision and accuracy were 10.0% RSD and within ± 8.5%; and the inter-assay precision and accuracy were 10.0% RSD and within ± 5.7% RE, respectively. The limit of quantitation of the hEGFl-48 and hEGFl-53 assay using 200 µL plasma per well is 7.8 and 62.5 pg/ml, respectively. These two ELISA methods are specific to hEGFs and do not cross-react with mouse EGF or other growth factors (TGF, TGF, PDGF, and FGF) or lymphokines (IL₁ and TNF). These validated methods have been routinely applied to assay of plasma samples from various pharmacokinetic studies in rats receiving intravenous hEGFs. Both assay methods were also adapted to assay endogenous hEGFs in biological fluids of different animal species. Conclusions. Two sensitive ELISA methods have been validated for quantitation of hEGFl–53 and hEGFl–48 in rat plasma. Their utility has been demonstrated in the application of assaying immunoreactive concentrations of exogenous and endogenous epidermal growth factors.     

合肥市科学岛社区高血压流行特征及影响因素调查

目的调查科学岛社区成人高血压流行特征及影响因素,探讨高血压社区管理的有效性。方法采取整群抽样及问卷调查的方法,对科学岛社区20周岁以上的常住人员进行高血压流行特征及影响因素的调查与分析。结果共调查2951人,资料完整者2646人参加体检,有421人患有高血压病,高血压病患病率为15.91%。结论本社区高血压知晓率、治疗率、控制达标率远高于全国平均水平,但控制达标率仍然较低,且伴有较高吸烟情况和较低的阿司匹林使用率。     

Synthesis,biological activity and conformational analysis of cyclic GRF analogs

A novel cyclic GRF analog, cyclo(Asp⁸-Lys¹²)-[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂, i.e. cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂, was synthesized by the solid phase procedure and found to retain significant biological activity. Solid phase cyclization of Asp⁸ to Lys¹² proceeded rapidly (～2h) using the BOP reagent. Substitution of Ala¹² with d -Ala² and/or NH₂-terminal replacement (desNH₂-Tyr¹ or N-MeTyr¹) in the cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ system resulted in highly potent analogs that were also active in vivo. Conformational analysis (circular dichroism and molecular dynamics calculations based on NOE-derived distance constraints) demonstrated that cyclo^8.12[Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ contains a long α-helical segment even in aqueous solution. A series of cyclo^8.12 stereoisomers containing d -Asp⁸ and/or d -Lys¹² were prepared and also found to be highly potent and to retain significant α-helical conformation. The high biological activity of cyclo^8.12[N-MeTyr¹,d -Ala²,Asp⁸,Ala¹⁵]-GRF(1-29)-NH₂ may be explained on the basis of retention of a preferred bioactive conformation.     

Synthesis and biological evaluation of mouse growth hormone-releasing factor

The recently described mouse growth hormone-releasing factor (mGRF) was synthesized by the solid phase procedure, purified by 2 stages of preparative high performance liquid chromatography and fully characterized. The biologic activity of the 42-amino acid peptide (H-His-Val-Asp-Ala-Ile-Phe-Thr-Thr-Asn-Tyr-Arg-Lys-Leu-Leu-Ser-Gln-Leu-Tyr-Ala-Arg-Lys-Val-Ile-Gln-Asp-Ile-Met-Asn-Lys-Gln-Gly-Glu-Arg-Ile-Gln-Glu-Gln-Arg-Ala-Arg-Leu-Ser-OH) was assessed in primary cultures of both mouse and rat anterior pituitary cells and compared to synthetic rat (rGRF) and human (hGRF) growth hormone-releasing factors. mGRF was equipotent to rGRF in mouse somatotrophs but slightly less potent in rat somatotrophs, while hGRF was 3-5 times less potent in both rodent species.     

Psychological characteristics and development of narcotic-addicted infants

This paper reviews neonatal and developmental behavioral characteristics of children born to heroin- and methadone-addicted women. Evidence of behavioral effects is clearer and more consistent for the neonatal period than in later infancy and childhood. It has not been shown that additional risk for psychological disturbance in childhood is contributed by passive addiction in utero. Methodologic problems in this literature are described and suggestions for strategies and directions in research are offered.