中国水仙水杨酸甲酯合成酶基因克隆与表达分析

为了解中国水仙(Narcissus tazetta var. chinensis)花香气形成的分子机理,以花发育形成相关基因的SSH文库获得的cDNA片段为基础,利用RACE技术从中国水仙花朵中克隆了水杨酸甲酯合成酶基因,命名为NtSAMT1 (GeneBank No. JX273470),其cDNA全长1323 bp,包含1个1131 bp完整阅读框架,编码376个氨基酸,具有Methyltransf_7 superfamily蛋白保守区,与仙女扇(Clarkia breweri) SAMT蛋白(1m6eX)的三维结构相似。系统进化树分析表明,中国水仙与粳稻的SAMT蛋白亲缘关系最近。原核表达结果表明NtSAMT1在大肠杆菌中能高效表达。半定量RT-PCR分析表明,NtSAMT1在花蕾期就有表达,第1天完全开放时各部位均有表达,以雄蕊与雌蕊的表达量最高,第8天已检测不到表达。     

大理和勐腊中缅树鼩血清及 肝差异代谢物的比较

为探究不同海拔地区中缅树鼩(Tupaia belangeri)血清和肝的差异代谢物及代谢通路变化,本研究采集大理(高海拔)和勐腊(低海拔)中缅树鼩分别12只和8只的血清和肝,采用非靶向代谢组气相色谱-质谱联用检测技术测定代谢物。结果表明,高海拔种群和低海拔种群相比,血清中一共有36种代谢物差异显著,其中,柠檬酸、葡萄糖、胆固醇等共32种代谢物的浓度上调,N-乙酰谷氨酸、葵酸、庚酸和对羟基苯甲酸这4种代谢物的浓度下调;高海拔种群肝差异代谢物和低海拔肝相比,一共有18种代谢物浓度差异显著,其中,苹果酸、核糖、葡萄糖等共10种差异代谢物浓度上调,谷氨酰胺、乙醇酸、硬脂酸等共8种差异代谢物浓度下调。高海拔种群与低海拔种群相比,血清中一共有76条代谢通路活性得分差异显著,其中,69条活性得分上调,7条活性得分下调;肝中一共有75条代谢通路活性得分差异显著,其中,43条活性得分上调,32条活性得分下调。以上结果说明,中缅树鼩在面对不同环境时,会调节不同组织中不同代谢通路(如三羧酸循环、糖酵解、脂类代谢和氨基酸代谢)的代谢物含量来适应环境,且血清比肝对环境变化更敏感。     

冷驯化条件下中缅树鼩脂肪组织代谢产物的变化

为探究冷驯化条件下中缅树鼩（Tupaia belangeri）白色脂肪组织（WAT）和褐色脂肪组织（BAT）的差异代谢物变化,本研究采集对照组和冷驯化28天组中缅树鼩的WAT和BAT,采用非靶向代谢组液相色谱—质谱联用检测技术分析其差异代谢物含量变化。结果表明,冷驯化组较对照组WAT中有7种差异代谢物显著上调;BAT中有25种差异代谢物,其中23种差异代谢物显著上调,2种差异代谢物显著下调,主要参与三羧酸循环、脂类代谢、氨基酸代谢和糖代谢等。以上结果说明中缅树鼩通过利用脂肪组织的不同差异代谢物来抵御低温环境,提高生存率。     

基于iTRAQ技术分析五倍子作用白念珠菌后的差异蛋白表达

为了探讨五倍子抗白念珠菌的作用机制,提取4 μg/mL五倍子提取液作用后的白念珠菌（SC5314）总蛋白,采用同位素标记相对和绝对定量（isobaric tags for relative and absolute quantitation,iTRAQ）蛋白质组学技术、液相色谱串联质谱（LC-MS/MS）技术分析和鉴定差异表达的蛋白质,并对差异表达蛋白进行生物信息学分析。经LC-MS/MS鉴定出3 721种蛋白质,其中,差异表达蛋白104种,包括57种表达上调蛋白和47种表达下调蛋白。通过生物信息学分析发现,上述差异蛋白参与了氧化还原反应、过氧化氢分解代谢以及能量代谢等生物学过程。研究表明,五倍子抗白念珠菌的作用机制可能是通过抑制氧化磷酸化,进而影响菌体能量代谢和物质生成,最终导致细胞结构与功能的改变。     

Ultrastructural and immunohistochemical demonstration of pancreatic polypeptide-containing F-cells in the stomach and pancreas of Tupaia belangeri

The gastro-entero-pancreatic (GEP) system in Tupaia belangeri contains a specific cell which reacts with antisera to pancreatic polypeptide (PP). The cells are scattered between exocrine pancreatic cells and also found in the pancreatic islets. Furthermore, they are also located in the fundic glands and to a very small extent in the corpus mucosa and in the glands of the upper duodenum. The cell reacting with antisera against PP is identified as the F-cell which has a specific ultrastructure especially with respect to its secretion. The present identification of the F-cell as the PP-cell in pancreas and stomach is discussed with respect to its possible functional implications.     

AR/PCC herb pair inhibits osteoblast pyroptosis to alleviate diabetes-related osteoporosis by activating Nrf2/Keap1 pathway

Osteoporosis is a prevalent complication of diabetes, characterized by systemic metabolic impairment of bone mass and microarchitecture, particularly in the spine. Anemarrhenae Rhizoma/Phellodendri Chinensis Cortex (AR/PCC) herb pair has been extensively employed in Traditional Chinese Medicine to manage diabetes; however, its potential to ameliorate diabetic osteoporosis (DOP) has remained obscure. Herein, we explored the protective efficacy of AR/PCC herb pair against DOP using a streptozotocin (STZ)-induced rat diabetic model. Our data showed that AR/PCC could effectively reduce the elevated fasting blood glucose and reverse the osteoporotic phenotype of diabetic rats, resulting in significant improvements in vertebral trabecular area percentage, trabecular thickness and trabecular number, while reducing trabecular separation. Specifically, AR/PCC herb pair improved impaired osteogenesis, nerve ingrowth and angiogenesis. More importantly, it could mitigate the aberrant activation of osteoblast pyroptosis in the vertebral bodies of diabetic rats by reducing increased expressions of Nlrp3, Asc, Caspase1, Gsdmd and IL-1β. Mechanistically, AR/PCC activated antioxidant pathway through the upregulation of the antioxidant response protein Nrf2, while concurrently decreasing its negative feedback regulator Keap1. Collectively, our in vivo findings demonstrate that AR/PCC can inhibit osteoblast pyroptosis and alleviate STZ-induced rat DOP, suggesting its potential as a therapeutic agent for mitigating DOP.     

饲料营养对饲血蚊传播HBV动物实验的影响

以含乙型肝炎病毒的人血饲喂三属蚊叮咬树，复制人类乙型肝炎病毒感染动物模型１～２个月后，由于饲料营养配比不当，树的出现消瘦、被毛蓬松、死亡率较高。经调整饲料有效营养配比，制成均衡营养颗粒饲料。提高了树乙型肝炎病毒感染模型的存活率，取得了较好的效果。     

中缅树鼩UCP2 cDNA核心片段的序列分析

解偶联蛋白2(UCP2)是近年新发现的一种解偶联蛋白,具有多种生物学活性,相关研究表明,UCP2可以抑制某些细胞(如免疫细胞等)线粒体活性氧的过量生成.本研究通过设计简并引物进行RT-PCR从中缅树鼩(Tupaia belangeri肝中获得UCP2基因cDNA核心序列.该片段长745 bp,推测氨基酸序列为248个氨基酸.结构功能分析发现,此段氨基酸序列具有2个线粒体内膜载体蛋白特征结构、5个跨膜α-螺旋结构域、1个嘌呤结合区域(PNBD)以及3个解耦联蛋白质的特征序列.中缅树鼩UCP2氨基酸序列与普氏野马(Equus caballus)、小家鼠(Mus musculus)、家犬(Canis lupus familiaris)、人(Homo sapiens)、褐家鼠(Rattus norvegicus)、豚鼠(Cavia porcellus)、苏门达腊猩猩(Pongo abelii)、加卡利 安鼠(Phodopus sungorus)和马铁菊头蝠(Rhinolophus ferrumequinum)UCP2进行比较,氨基酸同源性均在90％以上.同时,本研究通过MEGA5构建系统树,对UCP2分子进化特征进行了一定的探讨.     

树脑β－内啡肽能神经元胞体和纤维分布的研究

本文用免疫组织化学方法和免疫电镜方法对１４只树脑β－内啡肽能神经元胞体和纤维的分布及其在细胞器的定位进行了研究。结果表明，本文首次报道在Ｂｒｏｃａ斜角带观察到β－内啡肽免疫反应阳性神经元胞体，电镜观察到β－内啡肽免疫反应物质定位于大颗粒囊泡内的小颗粒上和粗面内质网上。下丘脑弓状核及其附近区域观察到β－内啡肽免疫反应阳性神经元胞体。在室周区、室旁核、第３脑室室管膜下层及室管膜上皮细胞间、内侧基底下丘脑及其外侧区、正中隆起内带和外带部可见到β－内啡肽免疫反应阳性纤维和串珠状的膨体。对β－内啡肽的释放途径及其调节因素作了探讨。