Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers

Human embryonic stem (hES) cells are typically maintained on mouse embryonic fibroblast (MEF) feeders or with MEF-conditioned medium. However, these xenosupport systems greatly limit the therapeutic applications of hES cells because of the risk of cross-transfer of animal pathogens. Here we showed that the bone morphogenetic protein antagonist noggin is critical in preventing differentiation of hES cells in culture. Furthermore, we found that the combination of noggin and basic fibroblast growth factor (bFGF) was sufficient to maintain the prolonged growth of hES cells while retaining all hES cell features. Since both noggin and bFGF are expressed in MEF, our findings suggest that they may be important factors secreted by MEF for maintaining undifferentiated pluripotent hES cells. Our data provide new insight into the mechanism how hES cell self-renewal is regulated. The newly developed feeder-free culture system will provide a more reliable alternative for future therapeutic applications of hES cells.     

Determination of cefazedone in human plasma by high performance liquid chromatography–tandem mass spectrometry: Application to a pharmacokinetic study on Chinese volunteers

A rapid, sensitive and simple high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method was developed for determination of cefazedone in human plasma using metronidazole as internal standard (IS). The chromatographic separation was achieved on an Ultimate XB-CN column (2.1 mm × 150 mm, 5 μm) with an isocratic mobile phase of acetonitrile and 20 mM ammonium acetate in 0.1% formic acid in water (15:85, v/v). Detection was performed using electrospray ionization in positive ion multiple reaction-monitoring mode (SRM), monitoring the transitions m/z 548.2 → 344.1 for cefazedone and m/z 172.2 → 128.1 for IS. Calibration curves were linear over a wide range of 0.20–401.12 μg/mL for cefazedone in plasma. The lower limit of quantification (LLOQ) was 0.20 μg/mL. The intra- and inter-day precisions were less than 7.2%. The average recovery of cefazedone was 90.8–91.0%. The validated method was successfully applied to the pharmacokinetic study of cefazedone in Chinese healthy volunteers following intravenous (IV) administration of 500, 1000 and 2000 mg cefazedone injection.     

干扰RNA的非靶免疫副作用及其siRNA序列设计与化学修饰

siRNA介导的RNA干扰技术已经成为基因功能研究和开展疾病治疗的有用工具.近年发现,siRNA在哺乳动物体内可激活天然免疫系统,诱导干扰素等炎症因子的分泌,并且可非特异性抑制某些非靶基因的表达,有可能极大限制RNA干扰技术的应用.进行高效特异性siRNA的设计和修饰,以保持或者增强siRNA的特异性靶基因沉默作用,又消除siRNA对机体的非靶免疫副作用,成为使siRNA安全有效应用于临床治疗的关键.     

基因组规模代谢网络模型构建及其应用

微生物制造产业的发展迫切需要进一步提高认识、设计和改造微生物细胞代谢的能力,以推动工业生物技术快速发展。随着微生物全基因组序列等高通量数据的不断积聚和生物信息学策略的持续涌现,使全局性、系统化地解析、设计、调控微生物生理代谢功能成为可能。而基于基因组序列注释和详细生化信息整合的基因组规模代谢网络模型(GSMM)构建为全局理解和理性调控微生物生理代谢功能提供了最佳平台。以下在详述GSMM的应用基础上,描述了如何构建一个高精确度的GSMM,并展望了未来的发展方向。     

聚乙烯醇生物降解研究进展

聚乙烯醇(PVA)是一种在纺织和化工行业中广泛使用的难降解的高分子聚合物。随着人们对纺织工业清洁生产的关注,如何在退浆工艺中就实现对PVA的生物降解、减少PVA废水的排放,并避免化学退浆过程中高温和氧化造成的棉纤维损伤,是近年来纺织生物技术领域的研究热点。由于PVA降解菌种类不多、培养周期长,PVA降解酶酶活不高、提取不容易等原因,使PVA的生化降解研究还局限在PVA降解菌的筛选、PVA降解酶的酶学性质研究等方面,PVA降解酶还未在纺织工业上得到应用。本文综述了近年来国内外在PVA降解菌筛选、PVA降解酶提取及酶学性质、PVA生化降解机理等方面的研究进展,并讨论了PVA生化降解研究中存在的问题及发展方向。     

Effects of size,neighbors, and site condition on tree growth in a subtropical evergreen and deciduous broad‐leaved mixed forest,China

Successful growth of a tree is the result of combined effects of biotic and abiotic factors. It is important to understand how biotic and abiotic factors affect changes in forest structure and dynamics under environmental fluctuations. In this study, we explored the effects of initial size [diameter at breast height (DBH)], neighborhood competition, and site condition on tree growth, based on a 3‐year monitoring of tree growth rate in a permanent plot (120 × 80 m) of montane Fagus engleriana–Cyclobalanopsis multiervis mixed forest on Mt. Shennongjia, China. We measured DBH increments every 6 months from October 2011 to October 2014 by field‐made dendrometers and calculated the mean annual growth rate over the 3 years for each individual tree. We also measured and calculated twelve soil properties and five topographic variables for 384 grids of 5 × 5 m. We defined two distance‐dependent neighborhood competition indices with and without considerations of phylogenetic relatedness between trees and tested for significant differences in growth rates among functional groups. On average, trees in this mixed montane forest grew 0.07 cm year^?1 in DBH. Deciduous, canopy, and early‐successional species grew faster than evergreen, small‐statured, and late‐successional species, respectively. Growth rates increased with initial DBH, but were not significantly related to neighborhood competition and site condition for overall trees. Phylogenetic relatedness between trees did not influence the neighborhood competition. Different factors were found to influence tree growth rates of different functional groups: Initial DBH was the dominant factor for all tree groups; neighborhood competition within 5 m radius decreased growth rates of evergreen trees; and site condition tended to be more related to growth rates of fast‐growing trees (deciduous, canopy, pioneer, and early‐successional species) than the slow‐growing trees (evergreen, understory, and late‐successional species).     

以胸痛为主要表现的胃食管反流病36例临床分析

目的：探讨以胸痛为主要表现的胃食管反流病的病因、临床特点及诊治方法。方法：选择本院近年来诊治的36例已排除心源性及食管器质性病变,诊断为胃食管反流病的胸痛患者,予以埃索美拉唑20mg,每日两次;吗叮林10mg,每日三次治疗8周,观察其疗效并分别于2周,4周,8周记录症状缓解情况。结果：经抗反流治疗8周后,30例（83.33%）胸痛完全消失,4例（11.11%）明显缓解,2例（5.56%）无效,总有效率达94.44%。胸痛症状在治疗2周后积分下降不明显,而治疗4周,8周后显著改善,与治疗前比较有显著性差异（P〈0.01）。结论：胃食管反流病病因复杂、临床表现多样易误诊漏诊,胃镜检查结合24小时食管动态ph监测,必要时给予质子泵抑制剂试验性治疗可提高本病的确诊率。     

不同预处理对大麦花药3种内源激素含量和过氧化物酶活性的影响

研究了大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅ Ｌ．）花药－花粉培养中不同预处理对花药内源激素（ＡＢＡ,ＩＡＡ,ＩＰＡ）含量和过氧化物酶活性的影响。结果表明：１． 经甘露醇预处理不同天数,同一种基因型的３种内源激素的变化和不同基因型的同一种内源激素的变化规律十分相似,均是在预处理初期含量急剧增加,最高值在０．５或１ ｄ 处。以后开始逐渐下降;最后,保持在一定的水平上。２． 在甘露醇预处理过程中,两种基因型花药过氧化物酶活性的变化规律也十分相似。在预处理前期（从开始到第３天）活性呈直线上升,第３天达到最大值。从第３天到第５天活性减弱。最后,活性又开始增强。３． 在低温预处理过程的初期（２～５ ｄ） 两种基因型花药过氧化物酶活性都出现第一个小峰。在第１４天（“Ｈａｒｒｉｎｇｔｏｎ”）或第２１天（“Ｉｇｒｉ”）又出现第二个峰值,但后者较高。在预处理的后期（从第２８天到第３５天）两种基因型花药过氧化物酶的活性又呈现上升的趋势。同甘露醇预处理后期变化一致     

Quality,bioactivity study,and preclinical acute toxicity,safety pharmacology evaluation of PEGylated recombinant human endostatin (M2ES)

Endostar, a potent endogenous antiangiogenic factor, is wildly used in clinics. However, it was easily degraded by enzymes and rapidly cleared by the kidneys. To overcome these shortcomings, PEGylated recombinant human endostatin was developed. In this study, the purity of M₂ES was evaluated by silver stain and reversed‐phase high‐performance liquid chromatography. Ultraviolet spectrum was used to examine the structural of M₂ES and endostar. The bioactivity and antitumor efficacy of M₂ES were evaluated using an in vitro endothelial cell migration model and athymic nude mouse xenograft model of a heterogeneous lung adenocarcinoma, respectively. A preclinical study was performed to evaluate the acute toxicity and safety pharmacology in rhesus monkeys. The purity of M₂ES was more than 98%; PEG modification has no effect on endostatin structure. Compared with the control group, M₂ES dramatically retards endothelial cell migration and tumor growth. After intravenous (IV) infusions of M₂ES at a dose level of three and 75 mg/kg in rhesus monkeys, there was no observable serious adverse event in both acute toxicity and safety pharmacology study. On the basis of the quality and bioactivity study data of M₂ES and the absence of serious side effect in rhesus monkeys, M₂ES was authorized to initiate a phase I clinical trial.