基于细菌群感效应人工构建分子开关

对生物体内已有的或者人工组装的生物合成途径进行优化操作涉及两个重要问题:代谢途径中关键酶的活性及蛋白表达水平。对于酶表达水平的研究,传统的做法是采用强启动子控制下的靶蛋白过量表达策略。靶蛋白的过量表达通常会导致细胞内积累大量的无活性包涵体,从而严重影响细胞的生理状态和相关生物途径的有效运转。针对这一问题,设计一种分子开关来精确调控生物合成过程中关键酶的表达水平,对于研究生物合成途径的代谢节律以及促进生物合成途径高效运转都具有重要的实用价值。基于细菌群落中普遍存在群感效应的基本原理并结合酶促催化的动力学特征,首先在大肠杆菌群落中建立信号分子高丝氨酸内酯(AHL)介导的细胞–细胞交流机制,将靶基因egfp置入到启动子PluxI的控制之下。在细胞生长过程中,产生的AHL累积到一定浓度启动靶基因表达。通过在细胞生长的不同阶段启动AHL降解酶AiiA的表达控制环境中信号分子AHL的浓度水平,从而控制靶基因egfp的转录效率,最终实现对靶蛋白EGFP表达水平的精确控制。通过检测细胞的生长状态、靶基因在mRNA水平、蛋白质水平的表达情况证明人工设计的分子开关可以便捷高效地控制靶基因表达水平,具有时空调节的严谨性。该分子开关有望广泛应用于代谢工程和合成生物学等研究领域中。     

云雾龙胆内生菌的分离鉴定及抗菌活性分析

从云雾龙胆(Gentiana nubigena)中分离到一株具有抗菌活性的内生细菌LD5,该菌株的发酵上清液对金黄色葡萄球菌和大肠杆菌的生长有显著的抑制作用。通过形态学、生理生化和16SrDNA系统进化分析,鉴定LD5为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。菌株LD5能合成黄酮类化合物、龙胆苦苷,以及一种与云雾龙胆醇提取物相似的代谢产物。     

CMTM6, the newly identified PD-L1 regulator,correlates with PD-L1 expression in lung cancers

Modulation of Immune check point regulators, especially the PD-1/PD-L1 axis, plays a critical role in successful management of a small proportion of lung cancer patients, but not so effective in the rest of lung cancer patients. A better understanding of immunotherapy non-responsive or resistant patients therefore warranted for future development of novel therapeutics. The newly identified regulator CMTM6 (CKLF-like MARVEL transmembrane domain containing 6) has been reported to serves as the stabilizer of PD-L1 and enhances the inhibitory effect of PD-L1 on immune system in both cell line and animal models, but its clinical relevance associated with PD-L1 is unknown and the current study is designed to address this question. The study using immunohistochemistry demonstrated that CMTM6 positivity from 15 out of 19 types of cancers with our in-house tissue microarray, and PD-L1 expression is always found only in CMTM6 positive cancers. CMTM6 and PD-L1 expression were analyzed in 81 lung cancer patient sample, and we observed that CMTM6 expression correlated with cancer histotypes and inversely correlated with cancer metastases, but not with patients’ age and gender. No PD-L1 expression was observed in negative CMTM6 samples. Higher expression PD-L1 is also associated with higher CMTM6 expression. In summary, CMTM6 expression is associated with PD-L1 expression, as well as lung cancer histotypes and metastasis. The results thus for the first time confirmed earlier reports on CMTM6/PD-L1 connection, from a clinical aspect of analysis.     

阿片类物质在中枢神经系统的免疫调控作用

内源及外源性阿片具有调节神经元与胶质细胞的功能,这些调节具有保护或损伤脑功能的双重作用。吗啡具有促进受病毒复制及继发感染的作用。另一方面,阿片受体中的ｋａｐｐａ受体可能具有保护神经元的作用。更深层次的研究应是了解阿片通过什么机制作用在胶质细胞和神经元上,藉此以促进研制出具有明显疗效的新药。     

关于几种避孕植物药的药理初筛及成份预试

本文对云南滇西地区民间用于避孕和绝育秘方中的槌果藤,野花椒寄生、花椒寄生、螃蟹树寄生、鸡矢藤及野花椒的醇提取物进行了小鼠最大耐受量测定及小鼠抗生育实验,结果表明,花椒寄生、螃蟹树寄生的醇提取物具有明显抗生育活性,槌果藤也具有一定的抗生育作用。此外,还对槌果藤、野花椒寄生进行了化学成份预试。     

三种草坪草的茎、叶解剖结构及其坪用性状

利用石蜡片法,对台湾草（Zoysia tenuifoia Willd.ex Trin.)、海雀稗（Paspalum vaginatum Sw.）、狗牙根[Cnodon dactylon(L.)Pers．]三种细叶暖季型草坪草的叶片有茎的解剖结构进行了研究。结果表明：叶片及茎的解剖结构与植物的耐旱性、耐践踏性和弹性等坪用特性有着密切联系。三种植物中,台湾草因叶片表皮细胞、泡状细胞、维管束鞘、机械组织有茎中纤维带的特征,而在耐旱性、耐踏踏性和弹性等坪用特性上表现出优于其他二种草坪草;狗牙根的耐旱性较强,耐踏踏性和弹性一般,海雀稗则在这几项坪用特征上均较弱。     

Improvement of xylose utilization in Clostridium acetobutylicum via expression of the talA gene encoding transaldolase from Escherichia coli

Clostridium acetobutylicum ATCC 824 was metabolically engineered for improved xylose utilization. The gene talA, which encodes transaldolase from Escherichia coli K-12, was cloned and overexpressed in C. acetobutylicum ATCC 824. Compared with C. acetobutylicum ATCC 824 (824-WT), the transformant bearing the E. coli talA gene (824-TAL) showed improved ability on xylose utilization and solvents production using xylose as the sole carbon source. During the fermentation of xylose and glucose mixtures with three xylose/glucose ratios (approximately 1:2, 1:1 and 2:1), the rate of xylose consumption and final solvents titers of 824-TAL were all higher than those of 824-WT, despite glucose repression on xylose uptake still existing. These results suggest that the insufficiency of transaldolase in the pentose phosphate pathway (PPP) of C. acetobutylicum is one of the bottlenecks for xylose metabolism and therefore, overexpressing the gene encoding transaldolase is able to improve xylose utilization and solvent production.     

Precise Reconstruction of Veins and Bile Ducts in Rat Liver Transplantation

Rat orthotopic liver transplantation (ROLT) remains a technically demanding procedure, especially regarding the reconstruction of the suprahepatic vena cava (SHVC). In this study, a new microsuture technique was developed for anastomosis of the SHVC, and a special single-groove cuff and blade-cut stent were introduced. With these modified techniques, we aimed to make a precise anastomosis of the SHVC and to provide optimal cuffs and stents for the reconstruction of the veins and bile ducts. According to different microsuture techniques for the SHVC and different types of cuffs and stents, three ROLT groups were created to compare the operation times and prognoses. Sham operations were performed as controls in the fourth group. The time expenditures with each step were compared among the transplantation groups. Biochemical parameters were tested at the end of a 1-month observation period. The short- and long-term survival rates of the transplantation groups were recorded and compared. Our new microsuture technique was faster than the conventional continuous suture technique for SHVC anastomosis (P < 0.05). The use of a single-groove cuff for reconstruction of the portal vein and the infrahepatic vena cava shortened the anastomotic time (P < 0.05). The use of blade-cut stents resulted in fewer biliary complications and better survival over the short and long terms (P < 0.05). Our new microsuture technique and the single-groove cuffs proved to be a precise method for venous reconstruction which shortened the anhepatic time and the anastomotic time significantly. The blade-cut stents apparently reduced the incidence of biliary complications. In summary, with this precise microsuture technique and delicate cuffs and stents, excellent long-term survival can be achieved easily and stably for ROLT.     

贵州赤水桫椤国家级自然保护区野猪危害特征与居民态度分析

野猪对农作物的危害是当前人与野生动物冲突的典型案例,分析其危害特征,掌握当地居民对野猪危害与保护管理的态度,对于自然保护区社区共管与野生动物保护管理决策尤为重要。对贵州赤水桫椤国家级自然保护区的调查研究发现,野猪一年四季均危害农作物,危害高峰期在 2-4 月与10-12 月,主要受作物收获期影响;农耕地在保护区的位置、距道路距离及距水源距离显著影响野猪对农耕地的危害,农耕地距道路(< 55 m)和水源越近(< 270 m),遭受危害程度越高。保护区的实验区和周边区域人居活动强烈,但野猪危害较其他区域严重,野猪危害农地时对水源因素较距居民点距离、距林缘距离、隐蔽因素更为敏感。70% 的当地居民认为野猪危害是由于种群数量过多引起。55% 的当地居民不赞同国家保护野猪的政策,62% 的赞同猎杀野猪控制其种群数量增长。受访者年龄与野猪危害程度两因素显著影响居民对野猪的容忍度,50 - 70 岁是当地社区居民的主要人群,也是态度中立及容忍度较高的人群。为有效减少野猪危害损失,调和人与野生动物冲突矛盾,建议保护区管理者根据野猪危害特征,帮助当地居民建立有效防控措施和有效快速的危害评估与损失补偿机制,同时根据当地居民的态度特征,做好保护政策的宣教工作。     

树载脂蛋白CIcDNA序列及其组织表达

分离提取树（ｔｒｅｅｓｈｒｅｗ,ＴＳ）肝组织ｍＲＮＡ,以此为模板,反转录构建了ＴＳ肝组织ｃＤＮＡ文库。利用制备的兔抗ＴＳ载脂蛋白ＣＩ多抗血清为探针筛选该文库,获得两个阳性克隆。测序及序列分析表明：两个克隆均为ＴＳａｐｏＣＩｃＤＮＡ基因。大的克隆由３８０个核苷酸构成,包括２１ｂｐ、９５ｂｐ组成的５′和３′非编码区,２６４ｂｐ组成的一个完整开放阅读框架,编码８８个氨基酸的ａｐｏＣＩ前体,含２６个氨基酸构成的信号肽和６２肽的成熟蛋白。其成熟肽长度与大鼠、小鼠和狗的相应结构一致,但较人和狒狒ａｐｏＣＩ的成熟肽多５个氨基酸。并对其功能域进行了初步预测。ＲＮＡ印迹显示ＴＳａｐｏＣＩｍＲＮＡ不仅主要在肝组织表达;而且不同于人和其他哺乳类动物,亦可少量在小肠表达。上述结果为进一步研究ＴＳａｐｏＣＩ基因组结构、功能和体外表达奠定了基础。