异丙酚对缺氧复氧鼠脑神经元NOS活性和HSP70家族表达的影响

目的：通过观察不同浓度异丙酚对原代培养胎鼠大脑神经元缺氧复氧过程的影响，探讨异丙酚的部分脑保护机制。方法：培养12天的胎鼠大脑神经元，随机分为四组：Ⅰ组正常对照组；Ⅱ组缺氧复氧组；Ⅲ组14μmol／L异丙酚组；Ⅳ组56μmol／L异丙酚组。Ⅲ组和Ⅳ组于缺氧前分别换入含有14μmol/L和56μmol／L异丙酚的培养液，随后缺氧30min。四组于缺氧后1h、2h、4h、6h和24h用分光光度法分别比较各组神经元NOS(一氧化氮合成酶)活性，同时四组于缺氧后1h、3h、6h、8h、24h、48h和72h分别用原位杂交法和免疫组化法观察Hsp70(热休克蛋白70)mRNA、Hsc70(热休克同源蛋白70)mRNA及Hsp70的表达。结果：①本研究的缺氧复氧过程可使神经元NOS活性增强，并可诱导Hsp70 mRNA、Hsc70mRNA和Hsp70表达，且表达高峰分别为24h、24h和48h；②在14μmol／L和56μmol／L异丙酚组，缺氧复氧鼠脑神经元NOS活性在复氧后4h内降低；③14μmol／L和56μmol／L异丙酚组的缺氧复氧鼠脑神经元Hsp70mRNA和Hsc70mRNA的表达高峰分别提前至8h和6h，而只有56μmol／L异丙酚组的缺氧复氧鼠脑神经元Hsp70的表达高峰才提前至24h。结论：异丙酚可抑制缺氧复氧引发的神经元NOS活性增强，同时异丙酚可从转录和翻译两个水平上促进鼠脑神经元热休克蛋白70家族的表达。

^(60)Co照射诱导急性髓系白血病细胞bcl-2基因家族表达变化分析

Ｂｃｌ－２是最重要的抗凋亡基因之一，既往对ｂｃｌ－２家族基因表达特点与肿瘤临床特征、疗效关系的认识，主要源于肿瘤细胞系，而以原代肿瘤细胞为研究对象，结合临床资料分析ｂｃｌ－２基因家族在肿瘤形成、治疗反应性中的意义探讨少见。为此，我们用６０Ｃｏ照射处理...

Opaque2转录因子对玉米α-醇溶蛋白基因家族成员表达的影响

醇溶蛋白是玉米主要的储存蛋白,其中α-醇溶蛋白含量最为丰富,对玉米籽粒品质有重要影响。本研究利用全基因组数据鉴定α-醇溶蛋白基因家族,并从基因保守motif、玉米籽粒不同发育时期基因的表达谱以及转录因子Opaque2对该家族成员表达的影响3个方面分析玉米α-醇溶蛋白基因家族。结果显示,玉米参考基因组中存在39个α-醇溶蛋白基因,其中z1A、z1B、z1C和z1D 4个亚族成员分别为12个、8个、14个和5个。基因保守motif分析发现该家族基因序列高度保守。表达谱分析结果显示该家族表达模式差异较大,其中z1A亚族在籽粒发育时期表达量最高。利用RNA-seq数据剖析在种子发育阶段Opaque 2(O2)野生型和突变体编码该家族成员的表达差异,结果显示O2突变体中z1C亚族成员表达显著下调,z1A亚族表达略有下调,z1B和z1D亚族成员表达量没有显著的变化。本研究在重新注释α-醇溶蛋白家族成员信息的基础上,对该家族的表达谱进行了分析,为今后玉米籽粒品质育种提供了必要遗传信息。

脑表达X连锁家族基因在泛癌诊断和预后预测中的价值

目的:探讨脑表达X连锁(BEX)家族基因在泛癌中的差异性表达情况,及其在泛癌诊断和预后中的价值。方法:于2022年4月10日自加州大学圣克鲁斯分校(UCSC Xena)在线数据库下载来自癌症基因组图谱(TCGA)数据库中33种不同肿瘤的转录组测序(RNA-seq)数据、生存数据、免疫亚型、基于RNA和DNA甲基化的干细胞评分。利用R软件(V.4.2.0)limma包分析TCGA数据库中BEX家族基因的表达情况,通过Wilcox检验分析比较BEX家族基因在泛癌组织及正常组织中的差异性表达;根据BEX家族基因表达水平的中位值将泛癌患者分为高表达组和低表达组,采用Kaplan-Meier生存分析评估BEX家族基因表达与患者总生存(OS)的关系,采用Cox比例风险模型分析BEX家族基因表达对泛癌患者OS的影响并绘制森林图。根据相关指数Cor值分析BEX家族基因的表达与泛癌患者肿瘤微环境、肿瘤干细胞相关性,Spearman相关法分析胃癌组织的肿瘤微环境、肿瘤干细胞指数及BEX家族基因表达的相关性。从CellMiner数据库下载不同肿瘤细胞株RNA-seq及药物敏感性数据,分析BEX家族基因的表达与药物敏感性之间的相关性。使用Kruskal检验分析泛癌及胃癌免疫亚型与BEX家族基因表达间的相关性。结果:TCGA数据库中,BEX3在泛癌组织高表达,BEX2和BEX4在泛癌组织中度表达,BEX1和BEX5在泛癌组织相对低表达;BEX2、BEX3与BEX4在胆管癌中表达最高,BEX5在子宫内膜癌中表达最高,BEX1在浸润性乳腺癌中表达最高;相较于正常组织样本,BEX家族基因在多种肿瘤组织中的表达上调或下调(均 P<0.05);生存分析表明BEX家族基因的表达与多种癌症OS相关,BEX1、BEX3、BEX4、BEX5高表达的部分肿瘤患者OS较低表达患者好,差异均有统计学意义(均 P<0.05),其余BEX家族基因高表达患者OS较低表达患者差,差异均具有统计学意义(均 P<0.05);Cox回归分析显示,BEX1高表达对于胃癌;BEX2高表达对于急性髓系白血病、胸腺癌、子宫内膜癌;BEX3高表达对于头颈鳞状细胞癌、肉瘤、胃癌、子宫内膜癌;BEX4高表达对于直肠腺癌、胃癌、子宫内膜癌;BEX5高表达对于肾嫌色细胞癌、胸腺癌,均为OS的危险因素(均 P<0.05)。BEX家族基因表达与多数肿瘤的基质评分呈负相关(均 P<0.05),与肿瘤干细胞评分呈正相关(均 P<0.05)。BEX家族基因表达与胃癌肿瘤干细胞评分呈负相关( P<0.05),与基质评分、总评分呈正相关(均 P<0.05)。CellMiner数据库不同肿瘤细胞株中,BEX家族基因可能与威罗菲尼(Cor=-0.368, P=0.004)、Kahalide f(Cor=-0.391, P=0.002)、O-6-苄基鸟嘌呤(Cor=-0.375, P=0.003)等药物的耐药性相关。BEX家族中各基因均与泛癌的免疫亚型有关,均在C5亚型中呈现高表达(均 P<0.05);对于胃癌,除BEX5基因外( P=0.24),其余基因均在C3亚型中高表达(均 P<0.05)。 结论:BEX家族基因的表达与泛癌患者的预后密切相关,且对于肿瘤微环境、肿瘤干细胞以及药物敏感性存在影响。BEX家族基因可能是泛癌诊断及预后的潜在生物标志物。

Cloning and Expression Analysis of the High Mobility B Group Genes in Arabidopsis thaliana

[Objective] The aim was to better research the function and action mode of High Mobility Group B （HMGB） proteins in higher plants. [Method] At2G33450,At5G23405 and At5G23420 genes of HMGB protein family in Arabidopsis thaliana were cloned by the use of RT-PCR method,and the expression of these three proteins in E.coli and Arabidopsis thaliana were detected by using SDS-PAGE,Northern blot and subcellular localization methods. [Result] The results showed that the molecular weights of the three proteins were 17.5,17.0 and 27.0 kD respectively,and the expression levels of the proteins in Arabidopsis thaliana were At5G23420At5G23405At2G33450. In addition,all the three proteins were located in nucleus. [Conclusion] The study will provide a basis for the further research on the biological function of HMGB proteins in higher plants.

Regulation of Bcl-2 Family in TIP30-Induced Apoptosis in Hepatoblastoma Cells

Objective: To investigate the role of TIP30 in the apoptotic signal pathwayin HepG2, and Hep3B and Hu-7 hepatoblastoma cell lines. Methods: In order to confirm whether TIP30conducted Bcl-2 family was involved in apoptosis signal pathway, MTT assay, in situ 3' end labellingof DNA assay and Western blot were carried out to detect the diverse apoptotic function of TIP30and the regulation of Bcl-2 family. Results: TIP30 induced apoptosis as evidenced by morphologicalchanges in hepatoblastoma cells, which was accompanied by up-regulating Bax and Bad proteins andstimulating them from cytoplasm to mitochondria, and down-regulating Bcl-xl, while it had no effecton the level of Bak protein. Conclusion: TIP30 induced apoptosis partly by modulating the proteinlevels of members of Bcl-2 family in hepatoblastoma cells. Elucidating the mechanism by which TIP30induces cell death might establish it as an anticancer factor.

Expansion of the Actin Gene Family in Amphioxus

Actins are a small family of ubiquitous proteins that are essential cytoskeletal components and are highly conserved during evolution. Actins are usually divided into two classes, the cytoplasmic and muscle actins, which have different functional roles. Here we systematically analyzed the actin genes in the genome of the primitive chordate amphioxus （Branchiostoma floridae）. We found that amphioxus contains more than 30 actin genes, many of which are linked. Phylogenetic analysis suggests the amphioxus actin genes have clearly undergone extensive expansion through tandem duplications. The actin genes＇ structure also varies a lot, containing 2 to 7 exons. We also cloned two muscle type of actin genes from the amphioxus （B. belcheri） and compared their expression patterns during early development. The slight difference in their expression suggests functional diversification of these actin genes. Our results shed light on the evolution both of actin genes themselves and their functional roles in chordate development.

Identification of Light-Harvesting Chlorophyll a/b-Binding Protein Genes of Zostera marina L.and Their Expression Under Different Environmental Conditions

Photosynthesis includes the collection of light and a/b-binding （LHC） proteins. In high plants, the LHC gene family constituting the light-harvesting complex ofphotosystems I and II. the transfer of solar energy using light-harvesting chlorophyll includes LHCA and LHCB sub-families, which encode proteins Zostera marina L. is a monocotyledonous angiosperm and inhab- its submerged marine environments rather than land environments. We characterized the Lhca and Lhcb gene families of Z. marina from the expressed sequence tags （EST） database. In total, 13 unigenes were annotated as ZmLhc, 6 in Lhca family and 7 in ZmLhcb family. ZmLHCA and ZmLHCB contained the conservative LHC motifs and amino acid residues binding chlorophyll. The average similarity among mature ZmLHCA and ZmLHCB was 48.91% and 48.66%, respectively, which indicated a high degree of diver- gence within ZmLHChc gene family. The reconstructed phylogenetic tree showed that the tree topology and phylogenetic relation- ship were similar to those reported in other high plants, suggesting that the Lhc genes were highly conservative and the classification of ZmLhc genes was consistent with the evolutionary position of Z. marina. Real-time reverse transcription （RT） PCR analysis showed that different members of ZmLhca and ZmLhcb responded to a stress in different expression patterns. Salinity, temperature, light intensity and light quality may affect the expression of most ZmLhca and ZmLhcb genes. Inorganic carbon concentration and acidity had no obvious effect on ZmLhca and ZmLhcb gene expression, except for ZmLhca6.

Identification and expression analysis of bovine ANGPTL gene family

Encoded by seven genes, angiopoietin-like （ANGPTL） family members structurally similar to the angiogenic regulating factor angiopoietin are known to possess biological activities in angiogenesis and metabolism. Here we reports for the first time the identification and expression analysis of all the seven members of bovine ANGPTL gene family, which were designated bANGPTL1 to bANGPTL7 in order. The seven bANGPTL genes consist of 4-9 exons, span 3800M-3000 bp and are located on different chromosomes. The deduced amino acid sequences of the members all possess an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain, both characteristics of angiopoietins. Phylogenetic analysis showed that the 32 identified ANGPTL homologs from 9 species could be classified into two major groups. Real-time quantitative PCR （Q-PCR） analysis revealed that the bANGPTL family members have different expression patterns. This study will be helpful for investigation on the biological role of the bANGPTL family in this economically important species. Furthermore, it provides an insight into the molecular evolution of the emerging ANGPTL family

The Dicistroviridae: An Emerging Family of Invertebrate Viruses

Dicistroviruses comprise a newly characterized and rapidly expanding family of small RNA viruses of invertebrates. Several features of this virus group have attracted considerable research interest in recent years. In this review I provide an overview of the Dicistroviridae and describe progress made toward the understanding and practical application of dicistroviruses, including (i) construction of the first infectious clone of a dicistrovirus, (ii) use of the baculovirus expression system for production of an infectious dicistrovirus, (iii) the use of Drosophila C virus for analysis of host response to virus infection, and (iv) correlation of the presence of Israeli acute paralysis virus with honey bee colony collapse disorder. The potential use of dicistroviruses for insect pest management is also discussed. The structure, mechanism and practical use of the internal ribosome entry site (IRES) elements has recently been reviewed elsewhere.

Identification and Expression Analysis of ARF Gene Family in Pepper

In order to understand the functions of ARF（Auxin Response Factor） gene family during the growth and development and under stress in pepper, 20 ARF genes were identified from the capsicum genome by bioinformatics and these genes were not well-distributed on 12 chromosomes of pepper. The results of the phylogeny relationship showed that ARF genes of pepper could be divided into Class I and II and could be further divided into seven sub-groups： Ia, Ib, Ic, Id, Ie, IIa and IIb. The results of gene structure showed that ARF gene family members were composed of 1-15 exons, and for each gene expression level of different tissues at different developmental stages revealed that the gene family had certain expression specificity. The qRT-PCR uncovered that high salt stress could significantly activate and inhibit the expression ofARF gene family.

A Pin gene families encoding components of auxin efflux carriers in Brassica juncea

Based on the sequence information of Arabidopsis PIN1, two cDNAs encoding PIN homologues from Brassica juncea, Bjpin2 and Bjpin3, were isolated through cDNA library screening. Bjpin2 and Bjpin3 encoded proteins containing 640 and 635 amino acid residues, respectively, which shared 97.5% identities with each other and were highly homologous to Arabidopsis PIN1, PIN2 and other putative PIN proteins. BJPIN2 and BjPIN3 had similar structures as AtPIN proteins. Northern blot analysis indicated that Bjpin2 was expressed in stem, leaf and floral tissues, while Bjpin3 was expressed predominantly in stem and hypocotyls. Two promoter fragments of pin genes, Bjpin-X and Bjpin-Z, were isolated by 'genome walking' technique using primers at 5'-end of pin cDNA. Promoter-gus fusion studies revealed the GUS activities driven by Bjpin-X were at internal side of xylem and petal; while those driven by Bjpin-Z were detected at leaf vein, epidermal cell and cortex of stem, vascular tissues and anther. Results of the pin genes with different expression patterns in B. juncea suggested the presence of a gene family.

Involvement of two glycoside hydrolase family 19 members in colony morphotype and virulence in Flavobacterium columnare

Flavobacterium columnare is the pathogenic agent of columnaris disease in aquaculture. Using a recently developed gene deletion strategy, two genes that encode the Glyco hydro_19 domain （GH19 domain） containing proteins, ghd-1 and ghd-2, were deleted separately and together from the F. columnare G4 wild type strain. Surprisingly, the single-, Aghd-1 and Aghd-2, and double-gene mutants, Aghd-1 Aghd-2, all had rhizoid and non-rhizoid colony morphotypes, which we named Aghd-1, Aghd-2, Aghd-1 Aghd-2, and NAghd-1, NAghd-2, and NAghd-1 Aghd-2. However, chitin utilization was not detected in either these mutants or in the wild type. Instead, skimmed milk degradation was observed for the mutants and the wild type; the non-rhizoid strain NAghd-2 exhibited higher degradation activity as revealed by the larger transparent circle on the skimmed milk plate. Using zebrafish as the model organism, we found that non-rhizoid mutants had higher LDs0 values and were less virulent because zebrafish infected with these survived longer. Transcriptome analysis between the non-rhizoid and rhizoid colony morphotypes of each mutant, i.e., NAghd-1 versus （vs） Aghd-1, NAghd-2 vs Aghd-2, and NAghd-1 Aghd-2 vs Aghd-1 Aghd-2, revealed a large number of differentially expressed genes, among which 39 genes were common in three of the pairs compared. Although most of these genes encode hypothetical proteins, a few molecules such as phage tail protein, rhs element Vgr protein, thiol-activated cytolysin, and TonB-dependent outer membrane receptor precursor, expression of which was down-regulated in non-rhizoid mutants but up-regulated in rhizoid mutants, may play a role F. columnare virulence.

EXPRESSION OF T CELL RECEPTOR V _α GENE FAMILIES IN INTRATHYROIDAL T CELLS OF CHINESE PATIENTS WITH GRAVES’ DISEASE

Patients with Graves’ disease (GD) have marked lymphocytic infiltration in their thyroid glands We examined the gene for the variable regions of the α chain of the Chinese T cell receptor(V α gene) in intrathyroidal T cells to determine the role of T cells in the pathogenesis of GD and offer potential for the development of immunotherapeutic remedies for GD Methods. We used the reverse transcription and polymerase chain reaction(RT PCR) to amplify complementary DNA(cDNA) for the 18 known families of the V α gene in intrathyroidal T cells from 5 patients with Graves’ disease The findings were compared with the results of peripheral blood T cells in the same patients as well as those in normal subjects Results. We found that marked restriction in the expression of T cell receptor V α genes by T cells from the thyroid tissue of Chinese patients with GD(P<0 001) An average of only 4 6±1 52 of the 18 V α genes were expressed in such samples, as compared with 10 4±2 30V α genes expressed in peripheral blood T cells from the same patients The pattern of expressed V α genes differed from patient to patient with no clear predominance Conclusions. Expression of intrathyroidal T cell receptor V α genes in GD is highly restricted suggesting the primacy of T cells in causing the disorders

增龄对大鼠腹主动脉壁连接蛋白mRNA表达的影响

连接蛋白（connexin，Cx）是一类多基因家族表达的保守蛋白。6个Cx单体在细胞膜上环绕排列形成具有6角形中空结构的寡聚体蛋白，称连接子。相邻细胞膜上的两个连接子呈30度角倾斜排列组成缝隙连接（gap junction）。缝隙连接是广泛存在于各种生物体内各个组织器官中的结构。目前在血管壁已确定的连接蛋白有：Cx37、40、43及45。大量临床资料表明，动脉粥样硬化、肿瘤及高血压等的发病率均随年龄增长而增高，其发生机制均涉及缝隙连接的作用。因此，了解Cx在不同阶段的变化具有重要的生理及病理意义。

肿瘤睾丸抗原PRAMEF12在肾细胞癌的表达研究

目的肿瘤睾丸抗原(cancer-testis antigens,CTAs)可限制性表达于睾丸生殖细胞及肿瘤细胞,并具有高免疫原性等特点,可激发肿瘤患者机体免疫应答,被认为是抗原特异性免疫治疗的潜在靶点。本实验分析和鉴定黑色素瘤优先表达抗原家族成员12(preferentially expressed antigen ofmelanoma family member 12,PRAMEF12)是否为一个新的肿瘤睾丸抗原,并探究其在肾癌组织中的表达。方法采用逆转录-聚合酶链反应(RT-PCR)分析PRAMEF12 mRNA在成人多器官正常组织及肾癌组织中的表达特点。蛋白印迹法(Western Blot)及免疫组织化学分析PRAMEF12蛋白在肾癌组织中的表达特点。运用统计学方法分析PRAMEF12蛋白的表达量和病例临床特点之间的关系。结果PRAMEF12显著表达于正常成人睾丸组织及肾癌组织中,但在其他正常组织中未见表达。PRAMEF12在70例肾癌组织中阳性表达率为60%(42/70),并与患者的年龄、性别及病理分期之间无显著统计学意义(P=0.758,P=0.837,P=0.457)。结论 PRAMEF12是一个新的肿瘤睾丸抗原,异常表达于部分肾癌患者,可能促进肾癌的发生与发展。

A genome-wide analysis of the ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES(AS2/LOB) gene family in barley(Hordeum vulgare L.)

ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES（AS2/LOB） genes are a family of plant specific transcription factors, which play an important role in the regulation of plant lateral organ development and metabolism. However, a genome-wide analysis of the AS2/LOB gene family is still not available for barley. In the present study, 24 AS2-like（ASL）/LOB domain（LBD） genes were identified based on the barley（Hordeum vulgare L.） genome sequence. A phylogenetic tree of ASL/LBD proteins from barley, Arabidopsis, maize, and rice was constructed. The ASL/LBD genes were classified into two classes, class I and class II, which were divided into five and two subgroups, respectively. Genes homologous in barley and Arabidopsis were analyzed. In addition, the structure and chromosomal locations of the genes were analyzed. Expression profiles indicated that barley HvA SL/LBD genes exhibit a variety of expression patterns, suggesting that they are involved in various aspects of physiological and developmental processes. This genome-wide analysis of the barley AS2/LOB gene family contributes to our understanding of the functions of the AS2/LOB gene family.

Genome-wide analysis and expression profiling of the phospholipase D gene family in Gossypium arboreum

The plant phospholipase D （PLD） plays versatile functions in multiple aspects of plant growth, development, and stress re- sponses. However, until now, our knowledge concerning the PLD gene family members and their expression patterns in cotton has been limited. In this study, we performed for the first time the genome-wide analysis and expression profiling of PLD gene family in Gossypium arboretum, and finally, a total of 19 non-redundant PLD genes （GaPLDs） were identified. Based on the phylogenetic analysis, they were divided into six well-supported clades （tx, 13/？, 8, ~, ~ and q~）. Most of the GaPLD genes with- in the same clade showed the similar exon-intron organization and highly conserved motif structures. Additionally, the chro- mosomal distribution pattern revealed that GaPLD genes were unevenly distributed across 10 of the 13 cotton chromosomes. Segmental duplication is the major contributor to the expansion of GaPLD gene family and estimated to have occurred from 19.61 to 20.44 million years ago when a recent large-scale genome duplication occurred in cotton. Moreover, the expression profiling provides the functional divergence of GaPLD genes in cotton and provides some new light on the molecular mecha- nisms of GaPLDcd and GaPLD62 in fiber development.

Temporal and Tissue-Specific Expression of Tomato 14-3-3 Gene Family in Response to Phosphorus Deficiency

Plants adapt to phosphorus （P） deficiency through a complex of biological processes and many genes are involved. Tomato （Solanum lycopersicum L. ＇Hezuo906＇） plants were selected to grown hydroponically to study the temporal and spatial gene expression patterns of the 14-3-3 gene family and their roles in response to P deficiency in tomato plants. Using real-time reverse-transcriptase polymerase chain reaction （RT-PCR）, we investigated the expression profiles in different tissues （root, stem and leaf） at short-term and long-term P-deficient stress phases. Results revealed that i） four members of 14-3-3 gene family （TFT1, TFT4, TFT6 and TFTT） were involved in the adaptation of tomato plants to P deficiency, ii） TFT7 responded quickly to P deficiency in the root, while TFT6 responded slowly to P deficiency in the leaf, iii） expression response of TFT4 to P-deficient stress was widely distributed in different tissues （root, stem and leaf） while TFT8 only displayed stem-specific expression, and iv） temporal and tissues-specific expression patterns to P deficiency suggested that isoform specificity existed in tomato 14-3-3 gene family. We propose that TFT7 （one member of e-like group in tomato 14-3-3 family） is the early responsive gene and may play a role in the adaptation of tomato plants to shortterm P deficiency, while TFT6 （one member of non-e group in tomato 14-3-3 family） is the later responsive gene and may play a role in the adaptation of tomato plants to long-term P deficiency.