A novel carbonyl reductase from Pichia stipitis for the production of ethyl (S)-4-chloro-3-hydroxybutanoate

An NADPH-dependent carbonyl reductase (PsCR) gene from Pichia stipitis was cloned. It contains an open reading frame of 849 bp encoding 283 amino acids whose sequence had less than 60% identity to known reductases that produce ethyl (S)-4-chloro-3-hydroxybutanoates (S-CHBE). When expressed in Escherichia coli, the recombinant PsCR exhibited an activity of 27 U/mg using ethyl 4-chloro-3-oxobutanoate (COBE) as a substrate. Reduction of COBE to (S)-CHBE by transformants in an aqueous mono-phase system for 18 h, gave a molar yield of 94% and an optical purity of the (S)-isomer of more than 99% enantiomeric excess.     

Repression of SIRT1 Promotes the Differentiation of Mouse Induced Pluripotent Stem Cells into Neural Stem Cells

The use of transplanting functional neural stem cells (NSCs) derived from induced pluripotent stem cells (iPSCs) has increased for the treatment of brain diseases. As such, it is important to understand the molecular mechanisms that promote NSCs differentiation of iPSCs for future NSC-based therapies. Sirtuin 1 (SIRT1), a NAD⁺-dependent protein deacetylase, has attracted significant attention over the past decade due to its prominent role in processes including organ development, longevity, and cancer. However, it remains unclear whether SIRT1 plays a role in the differentiation of mouse iPSCs toward NSCs. In this study, we produced NSCs from mouse iPSCs using serum-free medium supplemented with retinoic acid. We then assessed changes in the expression of SIRT1 and microRNA-34a, which regulates SIRT1 expression. Moreover, we used a SIRT1 inhibitor to investigate the role of SIRT1 in NSCs differentiation of iPSCs. Data revealed that the expression of SIRT1 decreased, whereas miRNAs-34a increased, during this process. In addition, the inhibition of SIRT1 enhanced the generation of NSCs and mature neurocytes. This suggests that SIRT1 negatively regulated the differentiation of mouse iPSCs into NSCs, and that this process may be regulated by miRNA-34a.     

颈内动脉支架置入术对颈动脉狭窄患者认知功能影响的研究

目的：研究颈动脉狭窄对认知功能的影响,探讨颈动脉支架置入术（CAS）患者术后不同时期认知功能的变化及其可能的机制。方法：选取2010年1月至2012年12月我院神经内科收住的急性脑梗死（前循环）患者75例,按颈内动脉狭窄程度,分成无狭窄组,轻度狭窄组（狭窄程度〈30％）,中度狭窄组（30-69％）和治疗组（70．99％）,前三组给予药物治疗,治疗组同时给予颈内动脉支架置入术,应用蒙特利尔认知评估量表（MoCA）、简易精神状态检查表（MMSE）、搭火柴测验（stickTest）,对患者进行认知功能评估,比较术前、术后不同时期认知功能的变化,同时利用SPECT／CT对治疗组20例患者术后局部脑血流改善状况进行评价。结果：治疗组和对照组在术前比较MoCA总分、MMSE评分、StickTest评分,发现治疗组评分低于对照组,且随着颈动脉狭窄程度的加重,认知功能受损越明显;治疗组在术后1周与术前比较,评分反而降低,差异有统计学意义;术后3个月、术后6个月与术前比较,评分均有所提高,差异有统计学意义;通过SPECT／CT对治疗组术前感兴趣区血流量与同侧小脑平均脑血流量比较,术后再与同侧小脑比较,结果显示术后脑灌注明显改善。结论：颈动脉狭窄与血管性认知功能障碍有关,而且认知功能障碍的程度与颈动脉狭窄程度呈正相关,颈内动脉支架置入术可最终改善认知功能,尤其表现在视空间／执行能力,延迟回忆能力,注意力等方面。     

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.     

不同硝化抑制剂对红壤氮素硝化作用及玉米产量和氮素利用率的影响

本研究分析添加不同种硝化抑制剂及其组合的高效稳定性氯化铵氮肥对红壤硝化作用、玉米产量和氮肥利用率的影响,旨在筛选出适合酸性红壤的高效稳定性氯化铵态氮肥。在氯化铵中分别添加硝化抑制剂2-氯-6-三甲基吡啶(CP)、3,4-二甲基吡唑磷酸盐(DMPP)和双氰胺(DCD)及其组合,制成6种高效稳定性氯化铵态氮肥,以不施氮肥(CK)和施氯化铵(N)为对照,进行等氮量玉米盆栽试验。结果表明: 与N处理相比,CP+DMPP和DMPP+DCD处理红壤中铵态氮含量提高56%～62%,显著高于CP、DMPP和DCD处理;土壤表观硝化率显著降低33%～34%。添加硝化抑制剂及其组合的6个处理均显著提高了玉米生物量和氮肥吸收利用率。与N处理相比,单独添加硝化抑制剂处理生物量均显著高于硝化抑制剂组合处理,平均提高1.3倍;添加DCD处理效果最显著,玉米籽粒产量、吸氮量和氮肥吸收利用率分别显著提高4.1、6.3和4.4倍。为了达到既能低成本又能提高产量和氮肥利用率的效果,在红壤上添加硝化抑制剂DCD是最佳选择。     

栀子提取物ZG对单纯疱疹病毒1型细胞吸附的影响

采用负染技术,借助高倍电子显微镜观察栀子提取物ZG作用后,病毒颗粒及其病毒吸附蛋白(virus attach-ment protein,VAP)的变化,考察药物是否直接改变或破坏病毒包膜蛋白的结构,使其失去感染性;采用异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记病毒,以肝素钠为参照,借助冷却慢扫描电荷耦合器件荧光成象技术,用Aquacomos软件进行图象分析,以探讨栀子提取物ZG不同加药方式对HSV-1吸附量的影响。结果表明栀子提取物ZG对HSV-1包膜表面的VAP无直接破坏作用,不影响病毒对Hep-2细胞的感染性;先加入肝素钠再进行病毒吸附及肝素钠病毒同时加入培养细胞这两种用药方式可明显减少细胞表面病毒的吸附量;栀子提取物ZG各种不同加药方式均能阻止HSV-1对Hep-2细胞表面的吸附,使病毒吸附量减少。     

茶园间作柑桔杨梅或吊瓜对叶蝉及蜘蛛类群数量和空间格局的影响

为评价茶园间作几种常见经济作物对重要害虫假眼小绿叶蝉及其主要天敌蜘蛛类群数量和空间格局的影响,遂选乌牛早品种纯茶园、乌牛早分别与柑桔、杨梅和吊瓜的间作茶园、以及安吉白茶与吊瓜间作茶园,2007年9月上旬—2008年12月下旬,每旬1次调查茶丛上、中、下层叶蝉和各种蜘蛛的数量。结果表明:(1)与纯茶园相比,间作茶园叶蝉种群数量和蜘蛛类群个体数量显著地增加,间作茶园蜘蛛种数显著地增加;(2)间作茶园茶丛上、中、下层叶蝉、蜘蛛个体数量分布明显区别于纯茶园茶丛上、中、下层叶蝉、蜘蛛个体数量分布;(3)茶丛上层的嫩梢是制作高档茶的原料,而纯茶园茶丛上层叶蝉虫口百分率为54.16%,间作茶园茶丛上层叶蝉虫口百分率皆减小,并且叶蝉高峰期间蜘蛛的跟随效应增强;(4)间作增加了经济收入并减少了防治次数。认为:(1)间作可在一定程度上调控叶蝉种群、蜘蛛类群的数量和空间格局;(2)间作可减轻叶蝉为害造成的产值损失,增强了茶园群落对于叶蝉的自然控制潜能。     

静脉注射41BBLGFP融合表达质粒体内表达及其生物学活性

通过RTPCR方法扩增小鼠41BBLcDNA,以pEGFPN1为载体,构建融合蛋白41BBLGFP重组表达质粒p41BBLGFP.采用基于流体力学原理建立的裸DNA体内转染技术,从小鼠尾静脉快速(15s)注射质粒p41BBLGFP进行体内转染.荧光显微镜观察组织切片,见小鼠肝、脾、肾及肺中均有报告基因GFP表达,尤以肝细胞中荧光最强.进一步用Western印迹和免疫组织化学染色法确定肝细胞表面表达41BBL,用Hsp70H22细胞抗原肽皮下免疫小鼠,同时尾静脉注射质粒p41BBLGFP,检测血清中IL2和IFNγ的分泌.结果显示,质粒注射联合免疫组小鼠血清IL2和IFNγ的浓度分别较生理盐水对照组增加了3倍和4倍;脾细胞对H22细胞的杀伤率则由单独免疫组的45.74%±3.27%增至86.74%±9.36%.结果表明,体内(主要在肝脏)转染质粒p41BBLGFP可以成功表达,表达产物具有41BBL的生物学活性,为进一步研究体内转染41BBL用于基因治疗奠定了基础.