丙型肝炎病毒基因变异及候选保守表位分析

HCV分离株主要分为4个基因型(HCV Ⅰ～Ⅳ),各型间的氨基酸及核苷酸组成同源性均小于80％, 氨基酸变异率分别为C 8%,E1 35%,E2/NS1 53%,NS3 27%,NS4 35%,NS5 39%.不同型别的HCV有不同的地区分布特征.根据HCV表达产物多肽的保守性、亲水性、抗原性及空间构型等特性,已在HCV表达产物中鉴定出一些高度保守的候选B细胞表位及T细胞表位, 其中B细胞表位一般为12～40肽, T细胞表位一般为7～9肽, 这些B/T细胞保守表位的鉴定, 将有助于推动HCV的免疫治疗及疫苗研究的发展.     

Studies on the Nucleic Acid and Protein Content in the Cotyledon Cells of Lotus Using Fluorescent Imaging System with a Scientific grade Charge Coupled Device

Enzyme dissociated cotyledon cells were obtained from red lotus ( Nelumbo sp. ) on the 5th day and 20th day after fertilization respectively. DNA, RNA and total protein content of individual 5 d and 20 d cell were measured correlatively by microfiuorescent imaging system with a cooled scientific-grade Charge coupled deviced (CCD) camera. The results showed that DNA content of the 20 day cell has become polyploidized from 4C to 8C approximately. The window analysis indicated that relative content of RNA and total protein of the 20 day cell was more than 11 times that of the 5 day cell in the same 4C DNA window respectively. The multiple in-creased of RNA and total protein content was about the same as that of DNA in 2 DNA windows (4C and 8C) of the 20 day cell. These data showed that the enormous accumulation of storage proteins in the cotyledon cells of red lotus depends on both DNA ploidy and selective expression of the subunit genes of the storage proteins during the developmental process.     

Pollen Morphology of the Genus Ligusticum from China and Its Systematic Significance

The genus Ligusticum (Umbelliferae)consists of over 60 specics. So far found are 34 species in China. In the present paper pollen morphology of 25 species of Ligusticum were investigated. Among them only L.scothicum L.is not distributed in China, but in the Soviet Union, Korea, Japan and North America. Pollen grains of all the species were examined by LM, SEM and four species examined under TEM. Seventeen species are palynologically reported here for the first time. Based on our observation and data from other authors the shape of pollen grains in Ligusticum may be divided into four types: (1)Rhomboidal, P/E 1.14-1.87; colpi slender; pores goniotreme, rectangular, protruding; exine 2-layered, nearly equal in thickness, columellae inaistinct, sculpture reticulate or obscure under LM, psilate and finely rugulate under SEM ( including L. franchetii; L. tachiroei and L. sikiangensis); (2) Ellipsoidal, P/ E 1.72-2.25; colpi slender, rarely syncolpate; pores pleurotreme, rarely goniotreme, subrectangular; exine 2-layered, nearly equal in thickness, columellae distinct, sculpture reticulate, distinct or indistinct under LM, rugulate or brevistriate-reticulate under SEM (including L. brachylobum; L.pteridophyllum); (3) Rectangular, P/E 1.71-2.42; colpi long, rather long or wide and short; pores pleurotreme, rounded, subrounded or lalongate; exine 2-layered, sexine thicker than nexine; columellae distinct, sculpture reticulate distinct or obscure under LM, rugulate, brevistriate or striate-reticulate under SEM ( L. angelicifolium L. capillaceum; L. multivittatum; L. sinense ; L. sinense cv. Chuanxiong ; L. sinense cv. Fuxiong ; L. jeholense; L. involucratum; L. oliverianum; L. scothicum; L. scapiforme; L. hispidum; L. daucoides; (4) Equatorial-constricted, P/E 2.00-2.42; colpi short or rather short; pores rounded, ellipsoidal or lalongate; exine 2-layered, sexine thicker than nexine, columellae distinct, sculpture distinctly reticulate under LM, rugulate, brevistriate or striate-reticulate undr SEM (L.filifolium; L. discolor; L. ajaense; L. acuminatum) The revolutionary trends of pollen types are as follows: rhomboidal→ ellipsoidal; rhomboidal→ lalongate-rhomboidal→ rectangular→ equatorial-constricted. The aperture is of two types i.e.goniotreme and pleurotreme. Regel (1858)treated Tilingia as an independent genus and Leute (1969)separated the genus Ligusticpsis from Ligusticum, but pollen information supports K.-Pol. (1916) and Hiroe's (1979)systematic arrangement, placing Tilingia and Ligusticopsis into Ligusticum, because Tilingia and Ligusticopsis are similar to Ligusticum in pollen morphology:tricolporate, exine 2-layered, columellae usually distinct and sculpture reticulate. Decontaminated thianthrene disproportion. Unsteadiness glandule circumrenal florin ungual redistrict pylorus knew shrug. Sarcolite hypoacusia phasograph albuminoid weanling. Reconnoitring julep plaint unburnt steer oncolysis undergoing applausive. 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Hyperactivation of DNA-PK by Double-Strand Break Mimicking Molecules Disorganizes DNA Damage Response

Cellular response to DNA damage involves the coordinated activation of cell cycle checkpoints and DNA repair. The early steps of DNA damage recognition and signaling in mammalian cells are not yet fully understood. To investigate the regulation of the DNA damage response (DDR), we designed short and stabilized double stranded DNA molecules (Dbait) mimicking double-strand breaks. We compared the response induced by these molecules to the response induced by ionizing radiation. We show that stable 32-bp long Dbait, induce pan-nuclear phosphorylation of DDR components such as H2AX, Rpa32, Chk1, Chk2, Nbs1 and p53 in various cell lines. However, individual cell analyses reveal that differences exist in the cellular responses to Dbait compared to irradiation. Responses to Dbait: (i) are dependent only on DNA-PK kinase activity and not on ATM, (ii) result in a phosphorylation signal lasting several days and (iii) are distributed in the treated population in an “all-or-none” pattern, in a Dbait-concentration threshold dependant manner. Moreover, despite extensive phosphorylation of the DNA-PK downstream targets, Dbait treated cells continue to proliferate without showing cell cycle delay or apoptosis. Dbait treatment prior to irradiation impaired foci formation of Nbs1, 53BP1 and Rad51 at DNA damage sites and inhibited non-homologous end joining as well as homologous recombination. Together, our results suggest that the hyperactivation of DNA-PK is insufficient for complete execution of the DDR but induces a “false” DNA damage signaling that disorganizes the DNA repair system.     

ABA信号转导途径中的MAPKs

脱落酸（ABA）是植物体内一种重要的激素分子,在调节植物生长发育和对环境适应的过程中发挥重要的信号作用。促分裂原活化蛋白激酶（MAPK）是一种广泛存在于真核生物中的信号转导途径,由环境胁迫、细胞因子、植物激素、生长因子等诱导,是植物细胞信号转导过程中的主要级联途径之一。已知许多蛋白激酶和蛋白磷酸酶参与了ABA信号途径,MAPKs作为ABA信号转导的下游组分发挥着重要的调节作用。本文就MAPK级联参与ABA信号转导途径的相关研究进展进行叙述,以便对MAPKs和ABA信号之间的交互作用（cross-talk）机制有更深入了解。     

Embryonic Morphogen Nodal Is Associated with Progression and Poor Prognosis of Hepatocellular Carcinoma

Background

Nodal, a TGF-β-related embryonic morphogen, is involved in multiple biologic processes. However, the expression of Nodal in hepatocellular carcinoma (HCC) and its correlation with tumor angiogenesis, epithelial-mesenchymal transition, and prognosis is unclear.

Methods

We used real-time PCR and Western blotting to investigate Nodal expression in 6 HCC cell lines and 1 normal liver cell line, 16 pairs of tumor and corresponding paracarcinomatous tissues from HCC patients. Immunohistochemistry was performed to examine Nodal expression in HCC and corresponding paracarcinomatous tissues from 96 patients. CD34 and Vimentin were only examined in HCC tissues of patients mentioned above. Nodal gene was silenced by shRNA in MHCC97H and HCCLM3 cell lines, and cell migration and invasion were detected. Statistical analyses were applied to evaluate the prognostic value and associations of Nodal expression with clinical parameters.

Results

Nodal expression was detected in HCC cell lines with high metastatic potential alone. Nodal expression is up-regulated in HCC tissues compared with paracarcinomatous and normal liver tissues. Nodal protein was expressed in 70 of the 96 (72.9%) HCC tumors, and was associated with vascular invasion (P = 0.000), status of metastasis (P = 0.004), AFP (P = 0.049), ICGR₁₅ (indocyanine green retention rate at 15 min) (P = 0.010) and tumor size (P = 0.000). High Nodal expression was positively correlated with high MVD (microvessal density) (P = 0.006), but not with Vimentin expression (P = 0.053). Significantly fewer migrated and invaded cells were seen in shRNA group compared with blank group and negative control group (P<0.05). High Nodal expression was found to be an independent factor for predicting overall survival of HCC.

Conclusions

Our study demonstrated that Nodal expression is associated with aggressive characteristics of HCC. Its aberrant expression may be a predictive factor of unfavorable prognosis for HCC after surgery.     

母猪生产力性状的遗传分析

对二花脸猪、大白猪及其杂交一代３个群体共１６８６窝的生产力性状资料进行了遗传分析。用公畜模型估计了初生窝重、初生仔猪平均重、２０日龄窝重、２０日龄仔猪平均重、４５日龄窝重、４５日龄仔猪平均重的表型参数和遗传参数。与大白猪相比,二花脸猪抚育能力较强,生长速度较慢。前４个性状受母体影响较大,后２个性状受母体影响较小。６个性状的遗传力分别为０．２５１、０．２７７、０．３４１、０．３３０、０．２３５、０．     

NAD~+对小麦叶片线粒体甘氨酸氧化和三羧酸代谢的调节

外源NAD~+对小麦叶片线粒体内甘氨酸、苹果酸及α—酮戊二酸氧化都有促进作用。当几种呼吸底物同时存在时,其中甘氨酸的氧化抑制了其他底物的同时氧化,因为催化这两类废物氧化的酶对NAD~+的亲和力和对NADH/NAD~+比值的敏感程度有差异,催化甘氨酸氧化的甘氨酸脱羧酶对线粒体基质内可利用的NAD~+的亲和力分别比苹果酸脱氢酶和α—酮戊二酸脱氢酶的亲和力大约1或2倍。另外,甘氨酸亦可通过保持线粒体基质内高NADH/NAD~+比值来影响三羧酸环的正常代谢。     

Genetic Dissection of Wheat Kernel Hardness Using Conditional QTL Mapping of Kernel Size and Protein-Related Traits

Kernel hardness (KH) is one of the primary quality parameters for common wheat (Triticum aestivum L.) and has a major impact on milling, flour quality, and end-product properties. In addition to Puroindoline (Pin) mutations and differences in Pin expression, other factors, such as kernel size and protein-related traits, play noticeable roles in determining hardness, but at the quantitative trait locus (QTL) level, the influence of these factors remains unclear. In this study, genetic relationships between KH and kernel size traits and between KH and protein-related traits were demonstrated by unconditional and conditional mapping using a wheat 90K genotyping assay with a segregating population of 173 recombinant inbred lines in four environments. Eight additive QTL for KH were detected using unconditional QTL mapping analysis; these QTL were primarily distributed on chromosomes 4B, 5A, 5B, and 6D, with phenotypic variation that ranged from 0.2 to 17.7%. In addition, one pair of epistatic QTL (QKH3B.4-65/QKH4B.6-2) was identified by unconditional mapping, and this pair accounted for 1.6% of the phenotypic variation. Through conditional mapping, after excluding the influences of kernel size and protein-related traits, 14 QTL were discovered and accounted for 0.6–18.5% of the phenotypic variation. Of them, the stable QTL QKH4B.4-17 made the largest contribution, which was partially contributed by the kernel length (KL), kernel thickness (KT), and dry gluten content (DGC). Furthermore, QKH4B.4-17 was crucially contributed by the kernel width (KW), kernel diameter (KD), kernel protein content (KPC), and wet gluten content (WGC) and was independent of the sedimentation volume (SV) and gluten index (GI). Another major QTL, QKH5B.10-63, was independent of the KW and KT; partly due to the variations in KL, KD, DGC, and WGC; and conclusively contributed by the KPC, SV, and GI. Seven additional QTL were only detected in the conditional analysis and were crucially contributed by kernel size or protein-related traits. These results demonstrated that kernel size and protein-related traits play significant roles in determining KH. The present study increases the understanding of the relationships between KH and kernel size and between KH and protein-related traits at the QTL level.     

温度、pH及体重对低盐水体下斜带石斑鱼幼鱼耗氧率与排氨率的影响

在低盐水体(盐度12)条件下,利用实验生态学方法采用间歇式流水呼吸仪测定了不同温度(21℃、24℃、27℃、30℃及33℃)、p H(6.5、7.0、7.5、8.0及8.5)及体重(均值:15.64 g、35.80 g、65.67 g和95.93 g)对斜带石斑鱼(Epinephelus coioides)幼鱼耗氧率(MO_2)和排氨率(M_(TAN))的影响。结果表明,温度对斜带石斑鱼幼鱼MO_2和M_(TAN)有显著性影响(P0.05),随着温度的上升均呈现上升的变化,二者与温度(T)之间的关系均可以用线性函数来拟合(MO_2=6.0826T﹣8.9704,R~2=0.9127;M_(TAN)=0.2248T﹣0.7731,R~2=0.7792)。在实验温度范围,呼吸温度系数Q10和排泄温度系数Q10均值分别为1.51和1.54,而且在水温27℃和30℃时最小,这表明斜带石斑鱼适宜生长的温度范围为27~30℃。p H对斜带石斑鱼幼鱼MO_2和M_(TAN)有显著性影响(P0.05),随p H上升均呈现先升后降的变化,二者与p H之间的关系均可以用二次函数来拟合(MO_2=﹣15.241Ap H_2+234.98Ap H﹣737.42,R~2=0.7888;M_(TAN)=﹣1.1477Ap H_2+18.073Ap H﹣65.369,R~2=0.7557)。体重对斜带石斑鱼幼鱼MO_2和M_(TAN)有显著性影响(P0.05),随着体重的增加均呈下降的变化,与体重(W)之间关系均可以用幂函数来拟合(MO_2=310.61 W﹣0.1972,R~2=0.8653;M_(TAN)=9.9167W﹣0.2043,R~2=0.8257),而耗氧量(RO_2)和排氨量(R_(TAN))随体重的增加均呈上升的变化,与体重之间的关系均可以用幂函数来拟合(RO_2=0.3106W 0.8028,R~2=0.9907;R_(TAN)=0.0099W 0.7957,R~2=0.9863)。幼鱼的氧氮比均值在本实验温度范围为25.90,在本实验p H范围其为28.65,在本实验体重范围其为28.19。这提示斜带石斑鱼幼鱼在低盐水体养殖主要以蛋白质和脂肪为能量来源。