Bioinformatic identification of genes encoding C1q-domain containing proteins in zebrafish

C1q is the first subcomponent of classical pathway in the complement system and a major link between innate and acquired immunities. The globular (gC1q) domain similar with C1q was also found in many non-complement C1q-domain-containing (C1qDC) proteins which have similar crystal structure to that of the multifunctional tumor necrosis factor (TNF) ligand family, and also have diverse functions. In this study, we identified a total of 52 independent gene sequences encoding C1q-domain-containing proteins through comprehensive searches of zebrafish genome, cDNA and EST databases. In comparison to 31 orthologous genes in human and different numbers in other species, a significant selective pressure was suggested during vertebrate evolution. Domain organization of C1q-domain-containing (C1qDC) proteins mainly includes a leading signal peptide, a collagen-like region of variable length, and a C-terminal C1q domain. There are 11 highly conserved residues within the C1q domain, among which 2 are invariant within the zebrafish gene set. A more extensive database searches also revealed homologous C1qDC proteins in other vertebrates, invertebrates and even bacterium, but no homologous sequences for encoding C1qDC proteins were found in many species that have a more recent evolutionary history with zebrafish. Therefore, further studies on C1q-domain-containing genes among different species will help us understand evolutionary mechanism of innate and acquired immunities.     

Analysis of metabolic flux in <Emphasis Type="Italic">Escherichia coli</Emphasis> expressing human-like collagen in fed-batch culture

Metabolic flux distributions of recombinant Escherichia coli BL21 expressing human-like collagen were determined by means of a stoichiometric network and metabolic balancing. At the batch growth stage, the fluxes of the pentose phosphate pathway were higher than the fluxes of the fed-batch growth phase and the production stage. After the temperature was increased, there was a substantially elevated energy demand for synthesizing human-like collagen and heat-shock proteins, which resulted in changes in metabolic fluxes. The activities of the Embden-Meyerhof-Parnas pathway and the tricarboxylic acid cycle were significantly enhanced, leading to a reduction in the fluxes of the pentose phosphate pathway and other anabolic pathways. The temperature upshift also caused an increase in NADPH production by isocitrate dehydrogenase in the tricarboxylic acid cycle. The metabolic model predicted the involvement of a transhydrogenase that generates additional NADH from NADPH, thereby increasing ATP regeneration in the respiratory chain. These data indicated that the maintenance energy for cellular activity increased with the increase in biomass in fed-batch culture, and that cell growth and synthesis of human-like collagen could clearly represent the changes in metabolic fluxes. At the production stage, more NADPH was used to synthesize human-like collagen than for maintaining cellular activity, cell growth, and cell propagation. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Enantioselective lipase-catalyzed kinetic resolution of N-(2-ethyl-6-methylphenyl)alanine

The enzyme (BSL2), a highly active lipase expressed from newly constructed strain of Bacillus subtilis BSL2, is used in the kinetic resolution of N-(2-ethyl-6-methylphenyl)alanine from the corresponding racemic methyl ester. Reaction conditions are optimized to enhance the enantioselectivity. The effects of various racemic alkyl esters, substrate concentration, operating temperature, pH of the aqueous medium and organic solvents on activity and enantioselectivity of BSL2 for kinetic resolution are also studied. A high enantiomeric ratio (E = 60.7) is reached in diisopropyl ether/water (10%, v/v) and the enantioselectivity is about 22-fold higher than that in pure buffered aqueous solution. The results show that the reaction medium greatly influences BSL2 reaction and its enantioselectivity in the hydrolysis of racemic methyl ester.     

石菖蒲与马蹄莲幼苗结构的比较解剖学研究

菖蒲属的分类和系统地位近两个世纪以来一直存在较大的争议。以菖蒲属的石菖蒲及马蹄莲属的马蹄莲为代表进行幼苗的比较解剖学研究,从幼苗结构的视角为菖蒲属独特的分类及系统地位提供证据。研究发现石菖蒲幼苗的子叶节区下部为较原始的“工”字形中始式的单中柱,而马蹄莲为散生中柱;石菖蒲根的维管柱为2—8原型星状中柱,马蹄莲为2—5原型星状中柱。石菖蒲根的内皮层细胞壁为马蹄形五面加厚;而马蹄莲为凯氏带四面加厚。石菖蒲细胞内的晶体为柱状晶,而马蹄莲为针晶。此外在子叶吸器的结构和其它贮藏物等方面也存在差异。据此认为菖蒲属应从天南星科中分出并单独成科;同时支持菖蒲属位于单子叶植物基部较孤立的系统地位。     

温室希蛛的生物学特性观察

温室希蛛Achaearanea tepidariorum在鲁东南沿海地区1年发生2代,但第2代不完整.越冬蜘蛛4月下旬出蜇,5月上旬开始产卵.卵期10 d,幼蛛期320 d,成蛛期76 d,越冬代成蛛寿命更长,雌雄性比为3∶1.成蛛日均食麦蛾4.9 头,是多种害虫的重要天敌.     

Amino acid residues in transmembrane domain 10 of organic anion transporting polypeptide 1B3 are critical for cholecystokinin octapeptide transport

Human organic anion transporting polypeptides (OATP) 1B1 and 1B3 are multispecific transporters that mediate uptake of amphipathic organic compounds into hepatocytes. The two OATPs contain 12 transmembrane domains (TMs) and share 80% amino acid sequence identity. Besides common substrates with OATP1B1, OATP1B3 specifically transports cholecystokinin octapeptide (CCK-8). To determine which structural domains and/or residues are important for the substrate selectivity of OATP1B3, we constructed a series of chimeric proteins between OATP1B3 and 1B1, expressed them in HEK293 cells, and determined rates of uptake of CCK-8 along with surface expression of the proteins. Replacing TM10 in OATP1B3 with TM10 of OATP1B1 resulted in a dramatically reduced degree of CCK-8 transport, indicating that TM10 is crucial for recognition and/or translocation of CCK-8. Using site-directed mutagenesis, we identified three key residues within TM10, namely, Y537, S545, and T550. When we replaced these residues with the corresponding amino acid residues found in OATP1B1, the level of CCK-8 transport was similarly low as for the replacement of the whole TM10. Kinetic experiments showed that the K m values for CCK-8 transport in the TM10 replacement and triple mutant were only 1.3 and 1.1 microM, respectively, as compared to 16.3 microM for wild-type OATP1B3. Similarly, the V max values dropped from 495.5 pmol (normalized mg) (-1) min (-1) for wild-type OATP1B3 to 13.3 and 19.0 pmol (normalized mg) (-1) min (-1) for the TM10 replacement and triple mutant, respectively. Molecular modeling indicated that two of the three identified residues might form hydrogen bonds with CCK-8. In conclusion, we have identified three amino acid residues (Y537, S545, and T550) in TM10 of OATP1B3 that are important for CCK-8 transport.     

套式PCR检测体系扩增效率的分析及对假阳性的控制

用扩增效率等量化的指标来控制PCR检测过程中出现的假阳性,调查了受假阳性污染的试剂在特定的PCR检测系统和环境中产生假阳性的PCR循环数界限,采用模板量与扩增倍数的线性回归关系分析系统的扩增效率并以此测定计算了两个不同循环数系统（C27＋15）和C27＋20）的检测界限分别为0．78kg和0．065fg,提出以此检测界限的1／2量作为假阳性耐受量,两个不同循环数系统的假阳性耐受量分别为580个和48个拷贝。     

牛磺酸和力竭运动对大鼠血液生物化学的影响

30只雄性Wistar大鼠按体重随要分为安静组、运动组、服牛磺酸运动组。后两组进行跑台运动至力竭后即刻,取血测定血糖和血乳酸以及血清MDA、LDH、GOT和CK。结果表明：与运动组相比较,服牛磺酸运动阻力竭运动时间有延长趋势,说明补充牛磺酸提高大鼠运动能力;与安静组相比,运动组力竭运动后血糖显著降低,血清LD、MDA、CK、GOT和LDH都显著升高,提示大鼠长时间运动由于中枢疲劳和外周疲劳而导致力竭;与运动组相比较,服牛磺酸运动组血糖降低和血乳酸升高的程度有减小的趋势,而血清MDA、CK、GOT、LDH升高的程度显著减小。研究结果显示,补充牛磺酸能提高机体的运动能力其主要是通过抵抗要体外周疲劳的产生而实现的。     

西藏拉萨地区三叠系诺利阶牙形石分带及其国际对比

麦龙岗剖面位于西藏拉萨地区达孜县麦龙岗村 (N2 9°5 6′0 6″ ,E91°2 7′5 5″)。麦龙岗组中共发现牙形石 7属 :Epigondolella ,Hindeodella ,Priniodella ,Xaniognathodus,Lonchodina ,Metalonchodina和Enantiognathus属。其中Epigondolella属有 9种 ,包括 1新种和 1未定种 ,它们是E .primitia ,E .spiculata ,E .cf.spiculata ,E .tozeri,E .violinformissp .nov .,E .cf.triangularisuniformis,E .postera ,E .bidentata ,E .sp .。基于本次发现的化石材料 ,对西藏林周地区麦龙岗组进行新的牙形石带划分。自下而上划分为 :?Epigondolellaprimitia ,E .spiculata ,E .tozer i,E .postera和E .bidentata等带。与毛力等 (1987)、Orchard(1983,1991,1994 )和Krystyn(1977)的划分方案进行对比和讨论