雷公藤多糖的提取和含量测定方法研究

目的建立雷公藤中多糖的测定方法,进行不同提取条件时,药物中多糖的含量比较.方法采用苯酚-硫酸比色法,于490nm处测定含量.结果该方法线性关系良好,平均加样回收率为100.5%,RSD＝1.23%.结论本法准确,可作为雷公藤中多糖的质量控制.     

0.8%黄精多糖滴眼液对干眼症的实验研究

目的观察0.8%黄精多糖滴眼液对实验性干眼症模型的治疗效果.方法将实验性干眼症日本大耳白兔随机分为空白对照组(模型组)、受试药物组(治疗组)和阳性药物对照组(对照组),分别用溶媒、0.8%黄精多糖滴眼液和泪然滴眼液治疗.观察Schirmer I试验和角膜结膜虎红染色点数,每周1次,共5周.结果各组模型动物Schirmer I试验滤纸湿长度和角结膜虎红染色点数分别在用药2周后和3周后差异有显著性,治疗组在用药2周后Schirmer I试验滤纸湿长明显增加,用药3周后虎红染色点数减少.结论 0.8%黄精多糖滴眼液对实验性干眼症有明显疗效.     

卡介菌多糖核酸的抗炎和抗过敏作用

目的　研究卡介菌多糖核酸 (BCG PSN)的抗炎、抗过敏作用。方法　观察对磷酸组胺所致豚鼠皮肤瘙痒的影响 ;采用二甲苯所致小鼠耳廓肿胀及角叉菜胶所致大鼠足跖肿胀实验观察抗炎作用 ;以 2 ,4 二硝基氟苯所致小鼠皮肤迟发型变态反应、大鼠同种被动皮肤过敏反应及小鼠异种被动皮肤过敏反应 ,探讨抗过敏作用。结果　豚鼠隔日肌内注射BCG PSN(0 1,0 2 ,0 4mg·kg-1) 3wk ,对磷酸组胺所致皮肤瘙痒无明显影响。小鼠隔日肌内注射BCG PSN(0 15 ,0 30 ,0 6 0mg·kg-1) 3wk ,可剂量依赖性地抑制二甲苯所致耳廓肿胀和耳异种被动皮肤过敏反应 ,高剂量时也可显著抑制 2 ,4 二硝基氟苯诱导的迟发型变态反应。大鼠隔日肌内注射BCG PSN(0 1,0 2 ,0 4mg·kg-1) 3wk ,可剂量依赖性地抑制角叉菜胶所致足跖肿胀和同种被动皮肤过敏反应。收稿日期 :2 0 0 3 -12 -2 4,修回日期 :2 0 0 4-0 2 -12作者简介 :刘桂珍 ( 1965 -) ,女 ,主管实验师 ,研究方向 :心血管药理学。Tel:0 73 1 2 3 5 5 0 77;胡长平 ( 1969-) ,男 ,博士 ,副教授 ,通迅作者 ,研究方向 :心血管药理学 ,Tel:0 73 1 2 3 5 5 0 77,E mail:huchangping2 0 0 1@yahoo .com .cn结论　BCG PSN对急性炎症、速发型变态反应和迟发型变态反应具有抑制作用。     

拟康氏木霉胞外多糖联合奥沙利铂可体外协同抑制结直肠癌细胞的增殖

目的 探讨拟康氏木霉胞外多糖（EPS）联合奥沙利铂（Oxa）用药对结肠癌细胞HCT116的协同抑制作用。方法 实验分为Control组（0 μg/mL）、Oxa组（8 μg/mL Oxa）、EPS组（100 μg/mL EPS）、EPS+Oxa组（8 μg/mL Oxa+100 μg/mL EPS）。CCK-8检测细胞活力，CompuSyn软件拟合Fa-CI曲线评价联用效果。流式检测凋亡和周期、划痕实验和transwell实验检测细胞迁移能力，Oxa和EPS相关基因与结直肠癌相关基因取交集进行PPI分析以及GO和KEGG富集分析。结果 Oxa单用或与EPS联用处理HCT116细胞，均可使HCT116细胞活力受到抑制，并呈现剂量与时间依耐性，两者联用有明显的协同作用（CI<1）。两药联用组的细胞总凋亡率与Oxa组以及对照组比较均显著升高（P<0.05）；联合用药组处于S期的比例高于其他各组。EPS和Oxa均具有抑制HCT116细胞迁移的作用，两者联用后抑制作用更为明显。KEGG分析显示主要涉及耐药、凋亡、血管新生等通路。结论 EPS联合奥沙利铂可协同抑制HCT116细胞增殖，促进该细胞凋亡和细胞S周期阻滞、抑制细胞迁移，铂耐药、PI3K-Akt、MAPK等信号通路发挥了关键作用。     

牛蒡多糖对K562细胞增殖的抑制及其机制的探讨

目的研究牛蒡多糖对K562细胞增殖的抑制作用并初步探索其机制。方法 MTT法检测牛蒡多糖对K562细胞增殖的抑制作用,RT-PCR检测BCL-2mRNA、Bax mRNA的表达。结果牛蒡多糖能明显抑制K562细胞的增殖;BCL-2基因表达下调,Bax的基因表达增多。结论牛蒡多糖对K562细胞增殖有抑制作用,其机制可能与BCL-2基因表达下调,Bax的表达上调有关。     

Lack of effectiveness of 13-valent pneumococcal conjugate vaccination against pneumococcal carriage density in Papua New Guinean infants

BackgroundPapua New Guinea (PNG) introduced the 13-valent pneumococcal conjugate vaccine (PCV13) in 2014, with administration at 1, 2, and 3 months of age. PCV13 has reduced or eliminated carriage of vaccine types in populations with low pneumococcal carriage prevalence, carriage density and serotype diversity. This study investigated PCV13 impact on serotype-specific pneumococcal carriage prevalence, density, and serotype diversity in PNG infants, who have some of the highest reported rates of pneumococcal carriage and disease in the world.MethodsNasopharyngeal swabs were collected at 1, 4 and 9 months of age from PCV13-vaccinated infants (n = 57) and age-/season-matched, unvaccinated infants (at approximately 1 month, n = 53; 4 months, n = 57; 9 months, n = 52). Serotype-specific pneumococcal carriage density and antimicrobial resistance genes were identified by qPCR and microarray.ResultsPneumococci were present in 89% of swabs, with 60 different serotypes and four non-encapsulated variants detected. Multiple serotype carriage was common (47% of swabs). Vaccine type carriage prevalence was similar between PCV13-vaccinated and unvaccinated infants at 4 and 9 months of age. The prevalence of non-vaccine type carriage was also similar between cohorts, with non-vaccine types present in three-quarters of samples (from both vaccinated and unvaccinated infants) by 4 months of age. The median pneumococcal carriage density was high and similar at each age group (~7.0 log₁₀ genome equivalents/mL). PCV13 had no effect on overall pneumococcal carriage density, vaccine type density, non-vaccine type density, or the prevalence of antimicrobial resistance genes.ConclusionPNG infants experience dense and diverse pneumococcal colonisation with concurrent serotypes from 1 month of age. PCV13 had no impact on pneumococcal carriage density, even for vaccine serotypes. The low prevalence of vaccine serotypes, high pneumococcal carriage density and abundance of non-vaccine serotypes likely contribute to the lack of PCV13 impact on carriage in PNG infants. Indirect effects of the infant PCV programs are likely to be limited in PNG. Alternative vaccines with broader coverage should be considered.     

Repeat pneumococcal polysaccharide vaccine in Indigenous Australian adults is associated with decreased immune responsiveness

BackgroundIndigenous adults residing in the Northern Territory of Australia experience elevated rates of invasive pneumococcal disease despite the routine use of 23-valent pneumococcal polysaccharide vaccine (23vPPV). We hypothesised that the limited protection from 23vPPV may be due to hyporesponsiveness as a result of vaccine failure from repeated vaccination. To explore this possibility, we evaluated the immune response to a first and second dose of 23vPPV in Indigenous adults and a first dose of 23vPPV in non-Indigenous adults.MethodsSerotype-specific IgG was measured by ELISA for all 23 vaccine serotypes at baseline and at one month post-vaccination. Individuals were considered to have an adequate immune response if paired sera demonstrated either: a four-fold rise in antibody concentration; a two-fold rise if the post vaccination antibody was >1.3 μg/ml but <4.0 μg/ml; or a post-vaccination antibody concentration >4.0 μg/ml for at least half of the serotypes tested (12/23). Our per-protocol analysis included the comparison of outcomes for three groups: Indigenous adults receiving a second 23vPPV dose (N = 20) and Indigenous (N = 60) and non-Indigenous adults (N = 25) receiving their first 23vPPV dose.ResultsAll non-Indigenous adults receiving a first dose of 23vPPV mounted an adequate immune response (25/25). There was no significant difference in the proportion of individuals with an adequate response using our definition (primary endpoint), with 88% of Indigenous adults mounted an adequate response following first dose 23vPPV (53/60) compared to 70% having an adequate response following a second dose of 23vPPV (14/20; p = 0.05). The risk difference between Indigenous participants receiving first dose compared to non-Indigenous participants receiving first dose was significant when comparing a response threshold of at least 70% (−27%, 95% CI: −43% to −11%; p = 0.01) and 90% (−38%, 95% CI: −60% to −16%; p = 0.006) of serotypes with a positive response.ConclusionIndigenous participants demonstrated a poorer response to a first dose 23vPPV compared to their non-Indigenous counterparts, with lower IgG following a second 23vPPV dose. These findings highlight the critical need to evaluate the efficacy of future pneumococcal vaccine programs in the Australian Indigenous populations that recommend repeated doses of 23vPPV.     

山茱萸多糖对衰老大鼠晶状体中Sirt1基因表达的影响

目的：初步探讨山茱萸多糖治疗年龄相关性白内障的可能机制，为其预防和治疗寻找新的药物和靶点。方法：选用60只昆明大鼠，按体质量随机分为3组，每组20只，分别为对照组（C组）、衰老模型组（A组）和山茱萸多糖给药组（D组）。在相同环境下，C组正常饮食，每日颈背部皮下注射0．9％氯化钠注射液，剂量计算参照其他给药组D-半乳糖给药量；A和D组正常饮食，每日颈背部皮下注射D-半乳糖100mg／（kg·d）；连续皮下注射45d后，C和A组每日按照等剂量灌药量灌服温开水；D组按照多糖0．4g／蛞灌服山莱萸多糖，连续灌服30d。试验终期颈椎离断术处死大鼠，4℃迅速取眼分离晶状体置于液氮备用。免疫组化方法检测各组间总超氧化物歧化酶（T-SOD）和总抗氧化能力（T-AOC）的变化，RT—PCR（逆转录cDNA的聚合酶链式扩增反应）检测各组Sirt1、p53和FOXO1 mRNA的表达变化。结果：A组和D组大鼠晶状体中Sirt1 mRNA的表达较C组均显著上升（P〈0．05、P〈0．01），且D组较A组显著上升（P〈0．01）；A组和D组大鼠晶状体中p53 mRNA的表达较C组显著上升（P均〈0．01），而D组较A组显著下降（P〈0．05）；A组和D组大鼠晶状体中FOXO1 mRNA的表达较C组显著上升（P〈0．05、P〈0．01），且D组较A组显著上升（P〈0．01）。另外A组和D组大鼠晶状体中T—SOD的活性较C组显著上升（P均〈0．01），且D组较A组显著上升（P〈0．01）；A纽和D组大晶状体中鼠T-AOC的活性较C组显著上升（P均〈0．01），且D较A组显著上升（P〈0．01）。结论：山茱萸多糖可能通过调节Sirt1的表达，从而调节下游基因p53和FOXO1的表达，最终抑制或延缓晶状体上皮细胞的凋亡，减缓年龄相关性白内障的进展。     

枸杞多糖对颅内动脉瘤大鼠NF-κB信号通路及血管内皮组织炎性损伤的影响

目的 探讨枸杞多糖(LBP)对颅内动脉瘤(IA)大鼠核蛋白因子-κB(NF-κB)蛋白通路及血管内皮组织炎性损伤的影响。方法 取SD雌性大鼠50只,采用切除双侧卵巢并结扎左侧颈总动脉及双侧肾动脉后支以构建IA模型,随机分为IA模型组、LBP低(5 mg/kg)、中(10 mg/kg)、高(20 mg/kg)剂量组,另取10只大鼠,只暴露卵巢、颈总动脉、双侧肾动脉后支,不进行摘除和结扎,作为对照组(Control组); 各组均于手术1周后,开始给药,Control组、IA模型组经灌胃给予生理盐水,LBP各剂量组灌胃给予相应剂量药物,1次/d,共给药30 d; 末次给药12 h后处死,取大鼠血液用酶联免疫吸附法(ELISA)检测血清炎性因子白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)及血管内皮损伤标志物血管内皮生长因子(VEGF)、血管内皮素(ET)水平; 分离取出大鼠脑动脉(Wills)环,在40倍显微镜下检查大脑动脉(Wills)环病理改变,并检测动脉瘤血管壁厚度及动脉瘤体积; 取脑动脉瘤血管组织,用苏木精-伊红染色(HE)检测动脉瘤血管组织病理变化; 蛋白免疫印迹法(Western blot)检测动脉瘤血管组织NF-κB,Toll样受体-4(TLR4)蛋白表达水平。结果 与Control组比较,IA模型组大鼠Willis环上凸起、管壁厚度和肿动脉瘤体积、动脉瘤内皮细胞空泡变性及炎性细胞浸润等病理损伤程度、血清IL-6,TNF-α,VEGF,ET水平、动脉瘤血管组织NF-κB和TLR4蛋白表达水平明显升高(P<0.05); 与IA模型组比较,LBP低、中、高剂量组大鼠Willis环上凸起、管壁厚度和动脉瘤体积、动脉瘤内皮细胞空泡变性及炎性细胞浸润等病理损伤程度、血清IL-6,TNF-α,VEGF,ET水平、动脉瘤血管组织NF-κB和TLR4蛋白表达水平明显降低(P<0.05),且LBP各剂量组上述指标水平呈剂量依赖性降低。结论 LBP可能通过抑制TLR4/NF-κB通路激活、降低炎症反应来减轻IA血管内皮组织损伤。