Production of mouse embryoid bodies with hepatic differentiation potential by stirred tank bioreactor

Embryonic stem (ES) cells can differentiate into functional hepatic lineage cells, which can potentially be used in biomedicine. To obtain hepatic lineage cells from ES cells, embryoid bodies (EBs) must be formed. In this study, we developed an EB formation system using a spinner flask for mass production of EBs. ES cells were inoculated into the spinner flask, where they formed EBs within 4 d. The EBs were then transferred into an attached culture for hepatic differentiation. To verify the hepatic lineage cells, albumin secretion and hepatic-specific gene expression were examined. We found that EBs formed by either the spinner flask or hanging drops exhibited similar albumin secretion potential and hepatic-specific gene expression. We conclude that the spinner flask method can be used to produce mouse EBs that can be used to mass produce hepatic lineage cells for use in biomedicine.     

三江平原湿地小叶章生产力模拟模型

利用气象台站的常规观测资料,依据植物生长模拟理论,以d为步长,建立了湿地小叶章(Deyeuxia angustifolia)植被生产力动态模拟模型。该模型包括3个子模块:1)光合作用与呼吸作用;2)干物质积累;3)同化物分配,主要考虑了温度和积水因子对植物生长的影响。并利用实测资料对该模型进行了检验,结果表明:小叶章地上活体、枯落物、茎、叶各器官枯落物的模拟值与实测值之间均呈极显著的线性相关(R²分别为0.98、0.99、0.99和0.92)。在相邻区域的检验结果也表明,季节性积水沼泽化草甸小叶章的地上生物量明显高于常年积水沼泽。两类湿地小叶章地上生物量的模拟值与实测值之间均呈极显著线性相关(R²分别为0.66和0.79)。相近区域长期定位观测点连续2年的模拟结果与实测值之间也具有极显著的线性相关(R²分别为0.97和0.76)。     

An efficient conversion of N-acetyl-D-glucosamine to N-acetyl-D-galactosamine and derivatives

2-Acetamido-2-deoxy-d-galactose (d-GalNAc) is an important monosaccharide widely distributed in nature. However, unlike its 4-epimer, the 2-acetamido-2-deoxy-d-glucose (d-GlcNAc), d-GalNAc is very expensive to obtain from commercial sources. Herein we report an efficient transformation that allows for the conversion of d-GlcNAc to a d-GalNAc derivative 7 in three steps and in 58.4-75% overall yields. The process was carried out on a greater than 20-g scale without the need of chromatography. The versatility of compound 7 was demonstrated by the synthesis of several useful monosaccharides and thiodisaccharides containing a d-GalNAc residue.     

Membrane-thinning effect of curcumin

Interaction of curcumin with lipid bilayers is not well understood. A recent experiment showed that curcumin significantly affected the single-channel lifetime of gramicidin in a 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayer without affecting its single-channel conductance. We performed two experiments to understand this result. By isothermal titration calorimetry, we measured the partition coefficient of curcumin binding to DOPC bilayers. By x-ray lamellar diffraction, we measured the thickness change of DOPC bilayers as a function of the curcumin/lipid ratio. A nonlinear membrane-thinning effect by curcumin was discovered. The gramicidin data were qualitatively interpreted by the combination of isothermal titration calorimetry and x-ray results. We show that not only does curcumin thin the lipid bilayer, it might also weaken its elasticity moduli. The result implies that curcumin may affect the function of membrane proteins by modifying the properties of the host membrane.     

Differences in cold tolerance and expression of two fatty acid desaturase genes in the leaves between fingered citron and its dwarf mutant

Fingered citron (Citrus medica L. var. sarcodactylis Swingle), a precious fruit ornamental plant, is sensitive to low temperature. Cold tolerance, evaluated by semi-lethal temperature, was lower in wild-type ‘Qingpi’ than in its mutant ‘Aihua’ trees obtained by γ-radiation. The full-length cDNAs of two genes encoding fatty acid desaturases involved in unsaturated fatty acid biosynthesis were isolated from the fingered citron leaves. The CmsFAD2 open reading frame (ORF) had 1,152?bp and was uninterrupted, encoding a polypeptide of 384 amino acids that showing 82% homology with the microsomal ω-6 desaturase CiFAD2 in Davidia involucrate. The CmsFAD8 ORF contained 1,373?bp and 7 introns, encoding a polypeptide of 458 amino acids showing 76% homology with the plastidial ω-3 desaturase BpFAD8 in Betula pendula. CmsFAD2 was expressed highly in leaves but low in roots and flowers, while CmsFAD8 was obviously expressed in three tissues. Compared with control group (28°C), the expression of CmsFAD2 and CmsFAD8 in leaves of two genotypes was significantly induced at 6°C. The increase of CmsFAD2 and CmsFAD8 was earlier and larger in cold-tolerant ‘Aihua’ than in cold-sensitive ‘Qingpi’. The linolenic acid content increased significantly in leaves of mutant ‘Aihua’ plants exposed to low temperature of 6°C. The results showed that a positive relationship between CmsFAD expression and genotype tolerance to cold may exist.     

猪瘟病毒NS3基因克隆、原核表达及间接ELISA方法初步建立

采用PCR方法从携带猪瘟病毒兔化弱毒(Hog cholera lapinized virus,HCLV)全长基因组cDNA的质粒pPOHCLV中扩增到长度为2000bp左右NS3基因序列,并将其克隆至原核表达载体pET-32a(+),构建成重组原核表达载体pETNS3。将pETNS3在大肠杆菌Rosetta(DE3)中进行优化表达,SDS-PAGE分析重组蛋白NS3主要以包涵体形式表达,分子大小约95kD。Western Blotting分析表明重组蛋白NS3具有免疫原性。采用Ni+亲和层析方法纯化得到重组蛋白NS3(90%)。以纯化的重组蛋白NS3为抗原初步建立了检测CSFVNS3抗体的间接ELISA方法,检测221份不同猪群和年龄猪的血清样品。检测结果与IDEXX公司CSFV-Ab检测试剂盒检测结果进行对比,阳性符合率为83.33%,阴性符合率为89.38%,总符合率为86.43%。30份存在差异的血清样品用间接免疫荧光法(Indirect immunofluorescence assay,IFA)进行检测,结果显示IFA检测结果与NS3间接ELISA和IDEXX公司CSFV-Ab检测试剂盒符合率分别为56.67%和43.33%。     

冷藏温度及时间对稻螟赤眼蜂品质的影响

稻螟赤眼蜂Trichogramma japonicum Ashmead是水稻螟虫的重要卵寄生性天敌,对水稻螟虫的自然种群有着一定的控制作用。本文以稻螟赤眼蜂预蛹期作为冷藏时期,在0、3、6、10℃4个温度和0、5、10、15、20、25、30 d 7个冷藏时间条件下,以羽化率、雌蜂百分率、寄生米蛾卵数量、雌蜂寿命、种群趋势指数作为指标,调查了冷藏对稻螟赤眼蜂品质的影响。研究表明,在4个温度下,稻螟赤眼蜂各指标均随着冷藏时间的增加而降低。各冷藏温度中,以0℃最为敏感,冷藏至5 d时,羽化率(38.11%)和寄生米蛾卵数量(19.4粒)显著低于未冷藏处理;冷藏至15 d时,雌蜂寿命由3.1 d降至1.9 d,种群趋势指数由23.36下降至1.07。10℃下,稻螟赤眼蜂各指标下降最为缓慢,冷藏至10 d时,羽化率(85.35%)和寄生米蛾卵数量(24.3个)与冷藏0 d的差异不显著;冷藏至15 d时,雌蜂寿命(2.7 d)与冷藏0 d的差异也不显著,种群趋势指数为12.52。综合各指标,工厂化繁殖稻螟赤眼蜂时,其适宜冷藏温度为10℃,冷藏时间为10 d。     

Mitochondrial DNA evidence of southward migration of Manchus in China

The Northeast area of China is a cross region between East Asia and Siberia. Although five populations from this area have been studied in maternal lineage, little is known about the genetics of other populations. In this study, forty-seven Manchu individuals were analyzed using a mitochondrial DNA marker, and fourteen mitochondrial DNA haplogroups, the representative haplogroups of east Eurasian, were identified. All analyses showed that Manchu were close to the neighboring populations such as Mongolian, Korean and northern Han Chinese, and were far from the other populations who lived in the cradle of Manchu, suggesting that the Manchu integrated gradually with natives following its southward migration.     

Dysfunction of pulmonary surfactant mediated by phospholipid oxidation is cholesterol-dependent

Pulmonary surfactant forms a cohesive film at the alveolar air-lung interface, lowering surface tension, and thus reducing the work of breathing and preventing atelectasis. Surfactant function becomes impaired during inflammation due to degradation of the surfactant lipids and proteins by free radicals. In this study, we examine the role of reactive nitrogen (RNS) and oxygen (ROS) species on surfactant function with and without physiological cholesterol levels (5–10%). Surface activity was assessed in vitro in a captive bubble surfactometer (CBS). Surfactant chemistry, monolayer fluidity and thermodynamic behavior were also recorded before and after oxidation. We report that physiologic amounts of cholesterol combined with oxidation results in severe impairment of surfactant function. We also show that surfactant polyunsaturated phospholipids are the most susceptible to oxidative alteration. Membrane thermodynamic experiments showed significant surfactant film stiffening after free radical exposure in the presence of cholesterol. These results point to a previously unappreciated role for cholesterol in amplifying defects in surface activity caused by oxidation of pulmonary surfactant, a finding that may have implications for treating several lung diseases.