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.     

新生儿抚触的研究进展

新生儿抚触作为儿科护理的新理念,是一种对新生儿健康十分有益的自然健康医疗技术。开展新生儿抚触对新生儿有特别重要的意义。抚触可以促进婴儿体格和智能的发育、增进睡眠、稳定情绪;抚触可以增强免疫力、促进食物的消化和吸收;抚触可以促进早产儿生长发育、新生儿缺氧缺血性脑病患儿的康复;抚触可以降低新生儿黄疸、避免发生胆红素脑病;抚触可促进母婴之间情感交流;抚触可利于产后妈妈的恢复。总之,无论是对家庭、社会、医院、还是对新生儿自身来讲,抚触都是非常有意义、重要且必要的,作为家长及医院工作人员都应该了解并掌握新生儿抚触的方法以及注意事项,因此,在临床工作和家庭中都应该积极主动的广泛开展新生儿抚触,目前各国都在研究并开展新生儿抚触这种护理技术,未来新生儿抚触将会被越来越多的家庭所认识并在医院中和家庭中广泛开展。     

热带林下人工种植阳春砂仁的生长与果实产量动态

调查了西双版纳不同海拔热带沟谷雨林和次生林下的阳春砂仁生长和果实产量动态．结果表明,西双版纳热带林下阳春砂仁自身年龄增长、林下光照不足和旱季水分胁迫影响阳春砂仁果实产量。随种植期增加,阳春砂仁果实产量和成熟植株密度降低．当林下光照水平在全日照的35％以下时,阳春砂仁果实产量随林下日照水平变化呈线性增加(P<0．05)。沟谷下方阳春砂仁果实产量显著高于上方(P<0.05)。海拔600～1000m,由于阳春砂仁的主花期从干热季3～4月推迟到雨季5月,果实产量显著增加,沟谷雨林和次生林下阳春砂仁果实产量差异不显著。因此,在海拔800～1000m沟谷中轮歇地次生林下有计划种植阳春砂仁,代替在沟谷雨林下种植阳春砂仁,既能解决沟谷雨林下光照和当地旱季水分不足对阳春砂仁果实产量的影响,又有利于热带沟谷雨林的保护。     

The thymic stromal cell line MTSC4 induced thymocyte apoptosis in a non-MHC-restricted manner

Mouse thymic stromal cell line 4 (MTSC4) is one of the stromal cell lines established in our laboratory. While losing the characteristics of epithelial cells, they express some surface markers shared with thymic dendritic cells (TDCs). To further study the biological functions of these cells, we compared the capability of MTSC4 with TDCs in the induction of thymocyte apoptosis, using thymic reaggregation culture system. Apoptosis of thymocytes induced by MTSC4 and TDCs was measured by Annexin V and PI staining and analyzed by flow cytometry. We found that MTSC4 selectively augmented the apoptosis of CD4^ 8^ (DP) thymocytes. This effect was Fas/FasL independent and could not be blocked by antibodies to MHC class I and class II molecules. In addition, MTSC4 enhanced the apoptosis of DP thymocytes from different strains of mice, which implies that MTSC4-induced thymocyte apoptosis is not mediated by the TCR recognition of self peptide/MHC molecules. In contrast to MTSC4, thymocyte apoptosis induced by TDCs was MHC-restricted. Thus, MHC-independent fashion of stromal-DP thymocyte interaction may be one of the ways to induce thymocyte apoptosis in thymus. Our study has also shown that the interaction of MTSC4 stromal cells and thymocytes is required for the induction of thymocyte apoptosis.     

双向凝胶电泳-飞行时间质谱技术鉴定小鼠胚泡黏附时子宫内膜nm23-M2／NDPK B的表达

运用双向聚丙烯酰胺凝胶电泳(2DPAGE)分析未交配小鼠子宫内膜和妊娠第五天(D5)小鼠子宫内膜胚泡黏附时植入位点及其旁组织蛋白质组。差异蛋白质组学显示,等电点(isoelectric point,pI)约7．1、分子量(molecular weight,Mw)约18kDa的蛋白质点在D5小鼠子宫内膜特别是植入位点表达上调。对此蛋白质点用基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption／ionization time of flying mass spectrometry,MALDI—TOF—MS)测定其胶内酶解后的肽质量指纹谱(Peptide Mass Fingerprint,PMF),经Mascot：Peptide Mass Fingerprint中SWISS-PROT数据库查询后,鉴定该蛋白质为鼠源性nm23-M2／NDPKB。RT—PCR和免疫组织化学结果也显示D5小鼠子宫内膜nm23-M2／NDPK B mRNA和蛋白表达明显增加。提示nm23-M2／NDPKB参与胚泡着床这一重要生命活动过程。     

血卟啉单甲醚在A549肺癌细胞内亚细胞分布的动态变化

目的:探讨血卟啉单甲醚(Hematoporphyrin monomethyl ether,HMME)在A549肺癌细胞内亚细胞分布的动态变化。方法:传代培养A549肺癌细胞,分别与光敏剂HMME孵育2 h和12 h。应用由荧光显微镜及电感耦合器材(Charge-coupled device,CCD)组成的高分辨率荧光显微成像系统,结合荧光探针标记技术,采用细胞器-细胞荧光强度比值法研究HMME在不同时间的亚细胞分布情况。结果:在2 h和12 h两个孵育时间点高尔基体的平均荧光强度比值(J1/J2)值都最高;随着孵育时间延长,A549细胞的四种细胞器J1/J2值都升高且溶酶体幅度最大。结论:孵育时间是影响HMME亚细胞分布的一个重要因素。随着孵育时间的延长,A549肺癌细胞各细胞器吸收HMME能力逐渐增强,尤以溶酶体显著。     

Critical role of interleukin-17/interleukin-17 receptor axis in mediating Con A-induced hepatitis

Concanavalin A (Con A)-induced hepatitis is thought to be a T-cell-mediated disease with active destruction of liver cells. Interleukin (IL)-17 is a cytokine produced principally by CD4(+) T cells. However, whether IL-17/IL-17 receptor (IL-17/IL-17R)-mediated responses are involved in T-cell-mediated Con A-induced liver injury remains unclear. In this study, we found that IL-17 expression was highly elevated in liver tissues during Con A-induced hepatitis. The increased levels of IL-17 were paralleled with the severity of liver injury reflected by Alanine aminotransaminase and histological assay as well as the secretion of tumor necrosis factor (TNF)-α and IL-6. Blockage of IL-17 significantly ameliorated Con A-induced hepatitis, while overexpression of IL-17 systemically resulted in massive hepatocyte necrosis in mice. Furthermore, overexpression of an IL-17R immunoglobulin G1 fusion protein significantly attenuated liver inflammation after acute Con A treatment. High expression of IL-17R on Kupffer cells was also observed along with the production of cytokines including TNF-α and IL-6. Inhibition of Kupffer cells by gadolinium chloride completely prevented Con A-induced liver injury and cytokine release. Finally, IL-17-expressing CD4(+) T and natural killer T cells were greatly increased in Con A-injected mice compared with that in controls. Overall, our results indicate that IL-17R signaling is critically involved in the pathogenesis in Con A-induced hepatitis, and blockade of IL-17/IL-17R signaling pathway may represent a novel therapeutic intervention in human autoimmune-related hepatitis.     

酵母表面展示脂肪酶合成己二酸二异辛酯

展示酶的酵母细胞既具有固定化酶的优点,又有制备简单、成本较低的特点.采用表面展示南极假丝酵母脂肪酶B (Candida antarctica lipase B,CALB)的毕赤酵母细胞催化合成己二酸二异辛酯(Diisooctyl adipate,DIOA),对该反应体系进行优化,并实现了初步工艺放大制备.经条件优化后,在10mL反应体系中,DIOA的产率可达85.0％.该工艺放大到200mL反应体系时,DIOA产率可达97.8％.经减压蒸馏,DIOA纯度可达到98.2％.该酵母表面展示脂肪酶在合成绿色润滑油己二酸二异辛酯中具有良好应用前景.     

芦荟苷A、B以及异芦荟色苷D的同时分离纯化

首次采用反相中压制备色谱从芦荟中一次分离制备得到芦荟苷A、B以及异芦荟色苷D。以库拉索芦荟丙酮粗提物为原料,采用中压制备色谱系统:SCO色谱柱(40 cm×26 cm,30～50μm),流动相甲醇-0.5%乙酸水(33∶67,V/V),流速20 mL/min,等度洗脱方式,柱温室温,检测波长254 nm,收集波长356 nm对芦荟样品进行分离制备,得到三种化合物单体。经高效液相色谱、紫外、红外、质谱及核磁共振等方法分析表明所得到的三种化合物分别是异芦荟色苷D、芦荟苷A和芦荟苷B,其纯度分别达到了98.0%,96.0%和98.9%。该方法简便,产品质量高,一次制备可以得到多种单体,为芦荟成分的测定与药理活性的研究提供了条件。