Community-wide convergent evolution in insect adaptation to toxic cardenolides by substitutions in the Na,K-ATPase

The extent of convergent molecular evolution is largely unknown, yet is critical to understanding the genetics of adaptation. Target site insensitivity to cardenolides is a prime candidate for studying molecular convergence because herbivores in six orders of insects have specialized on these plant poisons, which gain their toxicity by blocking an essential transmembrane carrier, the sodium pump (Na,K-ATPase). We investigated gene sequences of the Na,K-ATPase α-subunit in 18 insects feeding on cardenolide-containing plants (spanning 15 genera and four orders) to screen for amino acid substitutions that might lower sensitivity to cardenolides. The replacement N122H that was previously shown to confer resistance in the monarch butterfly (Danaus plexippus) and Chrysochus leaf beetles was found in four additional species, Oncopeltus fasciatus and Lygaeus kalmii (Heteroptera, Lygaeidae), Labidomera clivicollis (Coleoptera, Chrysomelidae), and Liriomyza asclepiadis (Diptera, Agromyzidae). Thus, across 300 Myr of insect divergence, specialization on cardenolide-containing plants resulted in molecular convergence for an adaptation likely involved in coevolution. Our screen revealed a number of other substitutions connected to cardenolide binding in mammals. We confirmed that some of the particular substitutions provide resistance to cardenolides by introducing five distinct constructs of the Drosophila melanogaster gene into susceptible eucaryotic cells under an ouabain selection regime. These functional assays demonstrate that combined substitutions of Q(111) and N(122) are synergistic, with greater than twofold higher resistance than either substitution alone and >12-fold resistance over the wild type. Thus, even across deep phylogenetic branches, evolutionary degrees of freedom seem to be limited by physiological constraints, such that the same molecular substitutions confer adaptation.     

雷公藤多苷治疗强直性脊柱炎的疗效及安全性分析研究

目的：研究雷公藤多苷治疗强直性脊柱炎的有效性及安全性。方法：选取我院2009年1月-2013年12月收治的强直性脊柱炎患者50例,采取随机数字表法分为两组,每组25例。对照组给予柳氮磺胺吡啶片治疗,试验组给予雷公藤多苷片治疗,观察与比较两组患者经不同药物治疗的临床疗效及安全性。结果：试验组优良率明显高于对照组（P〈0.05）,差异具有统计学意义。两组患者分别出现肝功能异常、腹泻及月经推迟的患者数无明显差异（P〉0.05）,差异不具有统计学意义。结论：雷公藤多苷片治疗强直性脊柱炎的临床效果显著,且不良反应的发生率无明显增加,提升了用药的安全性,值得广泛推广。     

HPLC法测定复方桔梗止咳片中甘草苷和甘草酸含量

目的：建立同时测定复方桔梗止咳片中甘草苷和甘草酸含量的HPLC检测方法。方法：采用Agilent C18（4.6 mm×150 mm,5μm）色谱柱;以乙腈-0.05%磷酸为流动相进行梯度洗脱;流速1.0 mL·min-1;检测波长237 nm;进样量：10μL;柱温30℃。结果：甘草苷、甘草酸分别在10.65~42.60 mg·L-1、97.30~389.30 mg·L-1范围内呈良好的线性关系,r分别为1.0000、1.0000;平均回收率（n=6）分别为98.80%（RSD=0.73%）、99.84%（RSD=0.10%）。测得6批复方桔梗止咳片,每片分别含甘草苷135.28μg、123.16μg、113.21μg、136.08μg、160.84μg、156.32μg,甘草酸441.36μg、405.86μg、364.69μg、446.93μg、518.22μg、509.65μg。结论：该方法简便,重复性好,可用于同时测定复方桔梗止咳片中甘草苷和甘草酸的含量。     

雷公藤多苷联合常规治疗对紫癜性肾炎患儿血清学指标的影响

目的:研究雷公藤多苷联合常规治疗对紫癜性肾炎患儿血清学指标的影响.方法:选择2011年3月～2014年6月期间在我院儿科接受治疗的紫癜性肾炎患儿进行研究,随机分为常规治疗组和雷公藤组,检测两组患者的免疫标志分子、细胞因子以及蛋白酶分子含量.结果:(1)免疫标志分子:与常规治疗组血液中免疫标志分子的含量比较,雷公藤组血液中CD28、B7-2、CD54、CD11a、CD11b、CD18的含量较低;(2)细胞因子:与常规治疗组血清细胞因子含量比较,雷公藤组血清中肿瘤坏死因子-a(TNF-α)、转化生长因子-β1(TGF-β1)、胰岛素样生长因子(IGF1)、胰岛素样生长因子结合蛋白IGFBP-3、肝细胞生长因子(HGF)和结缔组织生长因子(CTGF)含量较低;(3)蛋白酶分子:与常规治疗组血清蛋白酶分子含量比较,雷公藤组血清中基质金属蛋白酶2、9(MMP2、MMP9)、胱蛋白酶C(Cys C)含量较低,TIMP1、β1,3半乳糖基转移酶(β1,3GT)、Cosmc含量较高.结论:雷公藤多苷联合常规治疗有助于抑制自身免疫反应,减少紫癜性肾炎相关细胞因子产生,调节蛋白酶平衡,是治疗紫癜性肾炎的理想方法.     

Peptide-Bound Hydroxylysine and Large Polypeptides related to Collagen Synthesis

About 10% of the urinary hydroxylysine is linked to peptides. There is no significant difference between the excretion of peptide-bound hydroxylysine and hydroxylysine measured in the non-dialysable fraction of the urine. Non-dialysable hydroxylysine is highly glycosylated and contains mainly glucosyl-galactosyl-hydroxylysine. It is concluded that the urinary peptide-bound hydroxylysine is present in the non-dialysable collagen polypeptides. Their unusual carbohydrate composition is discussed.     

高效液相色谱法测定银杏叶及相关产品中总黄酮醇苷含量

目的:为银杏叶及相关产品中总黄酮醇苷含量测定建立统一的方法。方法:取适量样品加甲醇-25%盐酸溶液(4∶1)混合液25 ml,加热回流30 min,放冷,转移至50 ml容量瓶中,并加甲醇至刻度,摇匀,过0.45μm的滤膜,滤液进高效液相色谱分析。结果:总黄酮醇苷的三种成分线性关系良好,槲皮素、山柰素最低检出限为0.3μg,异鼠李素最低检出限为0.4μg,总黄酮醇苷最低检出限为2.51μg。结论:该方法操作简单,结果准确可靠。     

正交试验优化管花肉苁蓉水提取工艺

目的:优选管花肉苁蓉水提取最佳工艺。方法:以管花肉苁蓉中的苯乙醇苷提取量为检测指标,以料液比、提取时间、提取次数为考察因素,采用正交试验法进行优选。结果:管花肉苁蓉的最佳水提工艺为料液比1∶15,回流提取3次,每次1h。结论:本工艺成本低廉、安全性高、生产周期短、提取率高,对管花肉苁蓉制剂的大生产具有指导意义。     

雷公藤多苷联合醋酸泼尼松治疗中度活动型系统性红斑狼疮的疗效观察

目的探讨雷公藤多苷治疗中度活动型系统性红斑狼疮的疗效和安全性。方法 79例中度活动型系统性红斑狼疮患者随机分为观察组40例和对照组39例,在应用醋酸泼尼松基础上,观察组给予雷公藤多苷20mg/次,3次/d,口服;对照组给予甲氨蝶呤10mg/次,1次/周,口服。治疗6个月后评定2组疗效及不良反应发生情况,并比较2组治疗前、后24h尿蛋白、补体C3、系统性红斑狼疮疾病活动度（systemic lupus erythematosus disease activity index,SLEDAI）评分。结果观察组总有效率（87.5%）高于对照组（66.7%）（P〈0.05）;观察组治疗后24h尿蛋白、补体C3、SLEDAI评分分别为（0.65±0.12）g、（0.81±0.55）g/L、4.87±1.92,对照组分别为（0.97±0.39）g、（0.79±0.22）g/L、5.29±2.10,与治疗前（（2.16±0.36）g,（0.48±0.37）g/L,12.8±4.15;（2.08±0.21）g,（0.49±0.26）g/L,12.6±5.01）比较差异均有统计学意义（P〈0.05）;观察组治疗后24h尿蛋白水平低于对照组（P〈0.05）;观察组月经紊乱发生率（20.00%）高于对照组（2.56%）（P〈0.05）,恶心呕吐、上呼吸道感染等不良反应发生率比较差异均无统计学意义（P〉0.05）。结论醋酸泼尼松联合雷公藤多苷治疗中度活动型系统性红斑狼疮疗效优于联合甲氨蝶呤,且不良反应轻。     

白首乌C21甾体苷对Heps小鼠抑瘤作用和对造血功能影响

目的探讨自首乌C21甾体苷对肝癌Heps荷瘤小鼠的肿瘤抑制作用和对造血功能的影响。方法建立小鼠Heps肝癌移植肿瘤模型,随机分为模型组、白首乌C21甾体苷组、环磷酰胺组和联合组,进行抑瘤率计算;通过检测外周血象和骨髓细胞凋亡,观察其对荷瘤小鼠造血功能的影响。结果白首乌C21甾体昔有Heps肝癌抑制作用,且荷瘤小鼠体质状态不受影响,可促进荷瘤小鼠骨髓增殖,与化疗药合用可减轻化疗的骨髓抑制作用。结论白首乌C21甾体苷具有抗癌、刺激造血的双重作用,与化疗药合用可减轻其骨髓抑制作用。