Engineering stem cell niches in bioreactors

Stem cells, including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells and amniotic fluid stem cells have the potential to be expanded and differentiated into various cell types in the body. Efficient differentiation of stem cells with the desired tissue-specific function is critical for stem cell-based cell therapy, tissue engineering, drug discovery and disease modeling. Bioreactors provide a great platform to regulate the stem cell microenvironment, known as “niches”, to impact stem cell fate decision. The niche factors include the regulatory factors such as oxygen, extracellular matrix (synthetic and decellularized), paracrine/autocrine signaling and physical forces (i.e., mechanical force, electrical force and flow shear). The use of novel bioreactors with precise control and recapitulation of niche factors through modulating reactor operation parameters can enable efficient stem cell expansion and differentiation. Recently, the development of microfluidic devices and microbioreactors also provides powerful tools to manipulate the stem cell microenvironment by adjusting flow rate and cytokine gradients. In general, bioreactor engineering can be used to better modulate stem cell niches critical for stem cell expansion, differentiation and applications as novel cell-based biomedicines. This paper reviews important factors that can be more precisely controlled in bioreactors and their effects on stem cell engineering.     

Fatal poisoning of a Griffon vulture (Gyps fulvus) with methomyl

A young Griffon vulture died with signs of convulsion, vomiting, tremor, seizures propulsions and lack of co-ordination. Toxicological analysis of the stomach content revealed the presence of methomyl (9.3 mg/kg), methomyloxime (6.7 mg/kg) and deltamethrin (6.1 mg/kg). Concentrations of methomyl and its intermediate methomyloxime, combined with results of other analysis, indicate methomyl as the cause of death of this bird.     

接种马铃薯软腐病菌引起的烟草叶片爆发

用马铃薯软腐病菌处理烟草后,以化学法测定了叶片中O-2含量,发现在接种后1～14 h内只出现一次O-2爆发,不同于以往植物悬浮细胞研究中出现的两次爆发结果.接种软腐病菌后2 h和6 h烟草叶片O-2的电子自旋共振波谱图(electron spin resonance,ESR)显示了同样的一次O-2爆发结果.接种后烟草叶片的线粒体和叶绿体O-2爆发与叶片的同步,显示此两种细胞器参与烟草叶片在软腐病菌作用下的O-2发生.无光照叶绿体O-2的ESR谱线不出现,表明叶绿体O-2可能来源于光合电子传递链.马铃薯软腐病菌作用下烟草叶片的O-2爆发是多位点的.     

高温对萝芙木细胞膜、膜脂和抗坏血酸代谢造成的损伤

通过高温胁迫下萝芙木叶片电导率、膜脂过氧化物丙二醛(MDA)、自由基清除剂抗坏血酸的变化研究,探讨高温对萝芙木组织造成的损伤及自由基清除剂代谢的影响,从而揭示高温胁迫下萝芙木生长发育迟缓的原因.结果表明,热胁迫对萝芙木细胞膜造成损伤,使膜脂更多地被过氧化,抗坏血酸含量下降,致使萝芙木受到热损伤和氧化损伤,最终可能使其生长发育放缓,开花结果推迟.研究还表明,萝芙木经适当高温(37℃)热驯,能产生较多的抗坏血酸,以清除热胁迫诱导产生的氧自由基,降低氧化损伤程度.     

水稻品种抗褐飞虱不同生物型的稳定性

采用Tai氏方法分析估测12个水稻品种抗褐飞虱不同生物型的稳定性,以期为选育抗虫稳定性好的水稻品种提供较为有效的分析和监测方法.结果表明：光照强度、苗龄、施氮量对不同水稻品种对褐飞虱不同生物型的抗性表现及抗性稳定性有明显影响.抗褐飞虱生物型Ⅱ的品种中,RHT、RP1976-18-6-4-2、Ptb33的抗性较稳定,IR56的抗性不稳定,IR36、ASD7的抗性极不稳定;感褐飞虱生物型Ⅱ的品种中,TN1的感虫性稳定,桂华占、佛山油占、IR26的感虫性较稳定,国粳4号、Mudgo的感虫性不稳定.抗褐飞虱孟加拉型的品种中,RP1976-18-6-4-2、RHT、Ptb33的抗性不稳定,IR56的抗性极不稳定;感褐飞虱孟加拉型的品种中,桂华占、佛山油占、IR26的感虫性稳定;TN1、IR36的感虫性不稳定;国粳4号、Mudgo、ASD7的感虫性极不稳定.     

混交比例对桉树-乡土树种混交林优势树种叶片资源获取性状的影响

桉树-乡土树种混交林在提高林分生产力和生态系统功能等方面具有较大潜力。该研究以南亚热带4种桉树-乡土树种混交林(桉树与乡土树种混交比例分别为5:5、6:4、7:3、8:2)和桉树纯林为研究对象,研究了3种优势乡土树种华润楠(Machilus chinensis)、阴香(Cinnamomum burmannii)、灰木莲(Manglietia glauca)和速生树种尾叶桉(Eucalyptus urophylla)的叶片生理、结构和化学性状在不同比例混交林中的差异。结果表明,4优势造林树种的叶片性状存在明显的种间差异,其中灰木莲的比叶面积(SLA)、光合磷利用效率(PPUE)、单位质量叶片最大光合速率(A_mass)和蒸腾速率(T_mass)以及叶片养分含量最高,说明灰木莲采取资源获取型的生态策略;尾叶桉的SLA、A_mass、T_mass及叶片养分含量最低,但具有最高的PPUE,说明尾叶桉兼顾了资源获取型和保守型的物种特征。灰木莲与尾叶桉在SLA、A_mass、T_mass     

冬小麦小RNA高通量测序及生物信息学分析

东农冬麦1号是我国首例可在黑龙江高寒地区种植的强抗寒冬小麦(Triticum aestivum)品种。鉴于分蘖节的耐寒程度直接影响植株的安全越冬, 且MicroRNAs是一类生物体内普遍存在的非编码内源小分子RNA, 在植物的生长发育和适应胁迫等过程中具有重要作用, 该文基于HiSeq深度测序原理, 对5°C和–10°C胁迫下的东农冬麦1号分蘖节进行测序, 并开展生物信息学分析。结果表明, 在5°C文库中有2 229 955条特有小分子RNA序列, 并检测到35条已知miRNA, 属于30个miRNA家族, 其表达丰度介于1–173 810之间; 在–10°C文库中有3 721 449条特有小分子RNA序列, 并发现29条已知miRNA, 属于24个miRNA家族, 其表达丰度为1–105 868。靶基因预测结果表明, 5°C文库的30个miRNA家族中共预测到53个靶基因。功能分析结果显示, 这些基因主要参与转录调节、新陈代谢、胁迫响应和信号转导等过程。已知小麦miRNA的类型和表达丰度在5°C和–10°C两个文库中均有较大差异。     

Antibodies to gp350/220 enhance the ability of Epstein-Barr virus to infect epithelial cells

Epstein-Barr virus (EBV) is a persistent, orally transmitted herpesvirus that replicates in B cells and epithelial cells and is associated with lymphoid and epithelial malignancies. The virus binds to CD21 on B cells via glycoprotein gp350/220 and infects efficiently. Infection of cultured epithelial cells has not typically been efficient but can occur in the absence of gp350/220 and CD21 and in vivo is thought to be important to the development of nasopharyngeal carcinoma. We report here that antibodies to gp350/220, which inhibit EBV infection of B cells, enhance infection of epithelial cells. The effect is not mediated by Fc receptor binding but is further enhanced by antibody cross-linking, which may patch gp350/220 in the virus envelope. Saliva from EBV-seropositive individuals has similar effects that can be reversed by depletion of antibody. The results are consistent with a model in which gp350/220 interferes with the access of other important players to the epithelial cell surface. The results may have implications for the development of nasopharyngeal carcinoma in high-risk populations in which elevated titers of antibody to EBV lytic cycle proteins are prognostic.     

石斑鱼性反转相关基因 ECaM的克隆及表达特征分析

用甲睾酮片饲喂 2 ～ 4 龄赤点石斑鱼 (Epinephelus akaara) , 6 周后 90% 以上的雌鱼性逆转为功能性雄鱼 . 运用抑制性差减杂交技术 (SSH) ,结合 SMART cDNA 合成和 RACE-PCR 方法,从性反转雄鱼性腺中克隆到钙调蛋白基因 (ECaM). 该基因 cDNA 全长为 582 bp ,开放阅读框长 450 bp ,编码的蛋白质由 149 个氨基酸组成, 5 ′端非编码区 74 bp , 3 ′端非编码区 58 bp. 虚拟 RNA 印迹表明, ECaM 在性反转雄鱼性腺中表达,而在正常雌鱼性腺中表达微弱 . 各种组织的半定量 RT-PCR 显示, ECaM 在脑、心、肝、脾、肾都有转录,在精巢和下丘脑中表达水平较高,而在肌肉中表达甚弱 . 性反转不同时期性腺的半定量 RT-PCR 及蛋白质印迹表明,性逆转过程中性腺里 ECaM 的表达量逐渐增加 . 上述结果提示钙调蛋白可能在赤点石斑鱼性逆转过程中发挥着作用, ECaM 可能是促使石斑鱼由雌向雄转变的重要功能基因之一 .