维药蜀葵花化学成分的分离与结构鉴定(Ⅰ)

目的扩大维药蜀葵花(Althaea rosea(Linn.)Cavan.)的药用资源。方法采用正相硅胶、反相ODS、Sephadex LH-20等柱色谱及高效液相色谱手段对蜀葵花进行分离纯化,并通过理化性质与波谱分析方法鉴定化合物的结构。结果从蜀葵花体积分数95%乙醇提取物中分离得到14个单体成分,分别鉴定为东莨菪内酯(scopoletin,1)、3,5,7-三羟基色原酮(3,5,7-trihydroxychromone,2)、反式咖啡酸(trans-caffeic acid,3)、4-hydroxy-3-methoxycinnamyl-β-D-glucopyranoside(4)、3,4-dime-thoxycinnamyl-β-D-glucopyranoside(5)、对羟基苯甲醛(p-hydroxybenzaldehyde,6)、对羟基苯甲酸(p-hydroxybenzoic acid,7)、4-O-β-D-吡喃葡萄糖氧基苯甲酸(4-O-β-D-glucopyranosylbenzoate,8)、对羟基苯甲酸-β-D-吡喃葡萄糖酯苷(p-hydroxybenzoyl-β-D-glucopyranoside,9)、4-methoxybenzyl-β-D-glucopyranoside(10)、原儿茶酸(protocatechuic acid,11)、3,5-二甲氧基-4-羟基苯甲酸(3,5-di-methoxy-4-hydroxy benzoic acid,12)、腺苷(adenosine,13)和1H-吲哚-3-羧酸(1H-indole-3-carboxylic acid,14)。结论化合物2、4～6、8～10和14为首次从蜀葵属中分离得到,3、11和13为首次从蜀葵中分离得到,并首次报道了化合物5的核磁共振波谱数据。     

刺络拔罐法治疗玫瑰糠疹疗效观察

目的观察刺络拔罐法治疗玫瑰糠疹的临床疗效。方法将130例玫瑰糠疹患者随机分为治疗组和对照组,每组65例。治疗组采用刺络拔罐法治疗,对照组采用常规西医治疗。治疗2个疗程后进行疗效总结对照。结果治疗组总有效率为95.4%,对照组总有效率为67.7%,两组比较差异有统计学意义(P0.01),结果表明治疗组总有效率优于对照组。结论刺络拔罐法治疗玫瑰糠疹总有效率优于常规西医治疗。     

针刺配合中药外洗治疗玫瑰糠疹疗效观察

目的观察针刺配合中药外洗治疗玫瑰糠疹的临床疗效。方法将140例玫瑰糠疹患者随机分为治疗组73例和对照组67例。治疗组采用针刺配合中药外洗治疗,对照组采用药物治疗。治疗2个疗程后观察两组临床疗效。结果治疗组治疗1个疗程后总有效率为75.3%,对照组为62.7%,两组比较差异无统计学意义(P0.05)。治疗组治疗2个疗程后总有效率为91.8%,对照组为79.1%,两组比较差异具有统计学意义(P0.05)。结论针刺配合中药外洗是一种治疗玫瑰糠疹的有效方法。     

红景天苷诱导骨髓间充质干细胞向神经元样细胞分化中的Ca2＋/CaM信号通路

背景：课题组前期研究表明,红景天苷能诱导骨髓间充质干细胞向神经元样细胞定向分化,Ca2＋信号是实现其生物学信号传导的重要途径之一目的：探讨Ca 2＋/CaM 信号通路在红景天苷诱导骨髓间充质干细胞向神经细胞定向分化中的作用及机制。方法：实验分为空白对照组和红景天苷诱导组,红景天苷诱导组将不同质量浓度红景天苷（5,10,20,50,100 mg/L）作用骨髓间充质干细胞24 h和100 mg/L红景天苷作用骨髓间充质干细胞12,24,48,72 h。采用Western blot方法分别检测红景天苷诱导骨髓间充质干细胞后神经标志分子MAP2和Ca 2＋/CaM信号通路中关键蛋白CaM、CaMKⅡ的表达水平。另外实验设阻断剂组,分别加Ca2＋信号通路特异性阻断剂：500μmol/L EGTA （细胞外Ca2＋螯合剂）、1 mmol/L Nifedipine （L型Ca2＋通道阻断剂）、10 mmol/L LY294002（PI3K抑制剂）分别作用细胞30 min后,再加入100 mg/L红景天苷作用细胞24 h,采用Western blot方法检测阻断Ca2＋/CaM信号通路后NSE、CaM的表达情况。结果与结论：①红景天苷诱导后,MAP2的表达上调（P〈0.01）,说明红景天苷可诱导骨髓间充质干细胞分化为神经细胞。②不同质量浓度红景天苷诱导骨髓间充质干细胞24 h后,10 mg/L红景天苷组CaM、CaMKⅡ的表达与其他诱导组比较显著上调（P〈0.01）;同一质量浓度红景天苷诱导72 h后CaM、CaMKⅡ表达明显下调（P〈0.01）。③阻断细胞外Ca 2＋和PI3K信号通路后,NSE与CaM的表达水平较红景天苷诱导组上调（P〈0.05）。结果表明,红景天苷通过抑制Ca2＋/CaM信号通路实现骨髓间充质干细胞向神经细胞定向分化。     

Effect of the regimen of "Gaoshan Hongjingtian" on mechanism of Poly (ADP-ribose) polymerase regulation of nuclear factor kappa B in the experimental diabetic retinopathy

Background Poly (ADP-ribose) polymerase (PARP) plays an important role in the death of retinal capillary cells in diabetic retinopathy (DR), partly via its regulation of nuclear factor kappa B (NF-κB). The current study investigated the effect of the regimen of "Gaoshan Hongjingtian" on the mechanism of PARP regulation of NF-κB, and demonstrated the possible impact of the regimen of "Gaoshan Hongjingtian" and rhodiola sachalinensis on diabetic retinopathy. Methods Wistar rats were made diabetic with streptozotocin, then, were assigned to three groups. After 2 months, these diabetic rats treated with rhodiola sachalinensis or the regimen of “Gaoshan Hongjingtian”, or untreated. Analyses of expression levels of PARP, NF-κB and intercellular adhesion molecule-1 (ICAM-1) on the retinas of rats in different groups were performed by Western blotting and immunohistochemical assays, and mRNA levels of NF-κB and ICAM-1were determined by real-time PCR. In addition, the basement membranes of capillaries in the rats’ retinas were observed by using electron microscopy, and diabetes-induced capillary degeneration (ghost pericytes and acellular capillaries) were quantitated. Results From the third month after the injection, the diabetic rats were given daily with the regimen of “Gaoshan Hongjingtian”, rhodiola sachalinensis or tap water separately. The diabetic rats failed to gain weight when compared with normal age-matched ones, whereas their glycated hemoglobin levels were significantly increased. After 5 months, the mRNA levels of NF-κB and ICAM-1, the protein expression of PARP, NF-κB, and ICAM-1 were significantly increased in the retinas of diabetic rats in untreated group, compared to the nondiabetic controls. After 8 months, the number of degenerated retinal capillaries (ghost pericytes and acellular capillaries) was significantly increased in the diabetic rats in untreated group, compared to normal age-matched rats. The regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis inhibited diabetes-induced up-expression of PARP, NF-κB, and ICAM-1 in the retinas of diabetic rats with diabetic duration of 5 months. Treatment using the regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis significangtly inhibited increases in the number of acellular capillaries and pericyte ghosts and suppressed basement membrane thickening in the retinas of rats with diabetes for 8 months, as compared to control diabetic rats. Conclusions These results indicate that PARP plays a role in the pathogenesis of diabetic retinopathy. Rhodiola sachalinensis and the regimen of “Gaoshan Hongjingtian” may have acted on the mechanism of PARP regulation of NF-κB, which suppressed the expression of NF-κB and ICAM-1, and led to inhibition of retinal capillary degeneration.     

发酵红景天中红景天苷的含量测定

[目的]比较红景天发酵前后红景天苷的含量变化．[方法]采用HPLC法测定红景天苷的含量,色谱条件为KroasilC18色谱柱（4．6mm×250．0mm,5μm）;流动相为乙腈一水梯度洗脱,流动速度为1．0mL／min,检测波长为276nm,柱温为25℃,进样量为20gL．[结果]发酵红景天中红景天苷的含量为2．39％,红景天提取物为1．61％,通过微生物发酵使得红景天中活性成分红景天苷含量增加48．45％．[结论]本方法简便、快速、准确．     

玫瑰糠疹皮损血管细胞黏附分子-1的检测

目的探讨血管细胞黏附分子-1(VCAM-1)在玫瑰糠疹中的发病机制。方法采用免疫组化SP法检测玫瑰糠疹皮损血管内皮细胞VCAM-1的表达。结果皮损组血管内皮细胞VCAM-1的表达均高于正常皮肤组及玫瑰糠疹皮损周围正常皮肤组,玫瑰糠疹皮损周围正常皮肤组的表达高于正常皮肤组,差异均有显著性(P<0.05)。结论VCAM-1在玫瑰糠疹皮损血管内皮细胞上的高表达与真皮浅层血管周围淋巴细胞浸润有关,参与了玫瑰糠疹的炎症反应。     

窄谱中波紫外线治疗玫瑰糠疹的疗效

目的:观察窄谱中波紫外线(NB-UVB)治疗玫瑰糠疹的疗效。方法:40例确诊为玫瑰糠疹的患者进行NB-UVB治疗,5次1个疗程,共进行2个疗程。结果:NB-UVB治疗玫瑰糠疹治愈率达77.5%,有效率达100%。结论:采用NB-UVB治疗玫瑰糠疹安全有效,值得在临床进一步推广。     

低氧状态下红景天通过上调缺氧诱导因子1α促进人脐静脉内皮细胞肾上腺髓质素及其受体的表达

 目的  探讨低氧状态下红景天对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)中肾上腺髓质素(adrenomedullin,ADM)、降钙素受体样受体(calcitonin receptor-like receptor,CRLR) 、缺氧诱导因子1α(hypoxia-inducible factor 1 alpha,HIF-1α)表达的影响以及这一过程中HIF-1α的调控作用。方法HUVECs常规培养并设立正常氧、低氧对照组和低氧红景天组。CCK8法观察低氧状态不同浓度红景天干预不同时间的促细胞增殖作用;实时定量PCR和Western blot法测定ADM、CRLR、HIF-1α基因表达的情况;通过小干扰RNA方法敲低HIF-1α基因,实时定量PCR和Western blot法检测ADM、CRLR基因表达的变化。结果 红景天促进低氧状态下HUVECs增殖,促进低氧状态下内皮细胞ADM、CRLR以及HIF-1α的表达,HIF-1α siRNA可以显著抑制红景天对ADM及CRLR表达的促进作用。结论 低氧状态下红景天通过上调HIF-1α促进HUVECs ADM及其受体CRLR的表达。     

FOXP3在玫瑰糠疹、扁平苔藓、大斑块型副银屑病及蕈样肉芽肿中的表达及意义

目的探讨叉头状转录因子P3(FOXP3)在玫瑰糠疹、扁平苔藓、大斑块型副银屑病及蕈样肉芽肿中的表达及意义。方法采用免疫组织化学EnVision法检测21例玫瑰糠疹、19例扁平苔藓、17例大斑块型副银屑病、22例蕈样肉芽肿及10例正常人皮肤中FOXP3的表达。结果玫瑰糠疹、扁平苔藓皮损中FOXP3+淋巴细胞阳性率与正常人皮肤中的表达差异无统计学意义(P>0.05);大斑块型副银屑病与蕈样肉芽肿皮损中FOXP3+淋巴细胞阳性率均高于玫瑰糠疹、扁平苔藓及正常皮肤(P<0.05),但其二者之间的表达差异无统计学意义(P>0.05)。结论 FOXP3+淋巴细胞阳性率在炎症性皮肤病、有肿瘤发展倾向的炎症性皮肤病及皮肤淋巴瘤中的表达有逐步增高的趋势。其增多可能为肿瘤细胞的免疫逃逸提供了条件,进而促进了肿瘤的发生、发展。