一个新的水稻白化转绿突变体“白S”特性的研究

利用60Co-γ射线照射光温敏核不育系广占63-4S的干种子,以诱发其突变,从中选育出了一个白化转绿突变体白S。该突变体在前2叶除叶尖外完全白化,第3叶部分白化,从第4叶开始白化的叶片转绿,后期恢复成白绿相间条纹色。与亲本广占63-4S相比,白S的株高较矮,叶片叶绿素含量较低,每穗颖花数较多,但其它农艺和经济性状、育性、自交结实率等均无显著差异。白S所配6个组合在单株理论产量方面的超标优势为-17.6%~21.9%,平均为0.7%。对生产上如何利用白S的白化转绿特性有效地提高水稻杂交种的纯度进行了探讨。

中国大白猪SⅡ_1系主要繁殖性状的遗传参数估测

[目的]研究中国大白猪SⅡ1系繁殖性状的遗传规律。[方法]采用多性状动物模型DFREML方法,对中国大白猪SⅡ1系繁殖性状的遗传参数进行估测。[结果]总产仔数(TNB)、活产仔数(ANB)、初生窝重(LWB)和21日龄窝重(LW21)的遗传力估计值分别为0.11、0.09、0.14和0.05。各性状间(TNB/ANB、TNB/LWB、TNB/LW21、ANB/LWB、ANB/LW21、LWB/LW21)的遗传相关分别为0.91、0.76、0.65、0.85、0.59和0.38。[结论]中国大白猪SⅡ1系主要繁殖性状的表型参数均已达到或超过选育目标。

中国大白猪S_Ⅱ系繁殖性状的遗传参数估测

应用多性状动物模型DFREML方法估计了中国大白猪SII系繁殖性状的遗传参数。结果表明,窝均产仔数、产活仔数、初生窝重、20日龄窝重的遗传力估计值分别为0.06、0.10、0.07和0.15。20日龄窝重(LW20)的母体效应为0.20,而其他性状的母体效应均较低。各性状TNB/ANB、TNB/LWB、TNB/LW20、ANB/LWB、ANB/LW20、LWB/LW20的遗传相关分别为0.94、0.81、0.74、0.88、0.71和0.53。

中国大白猪SⅡ_1系主要生长和胴体性状的遗传参数估测

应用多性状动物模型DFREML方法,估计了中国大白猪SⅡ1系生长和胴体性状的遗传参数。结果显示,生长性状中的平均日增重(ADG)、100kg体重日龄(AGE)、达100kg体重的背膘厚(BF)、饲料转化率(FCR)的遗传力估计值分别为0.26、0.21、0.45、0.39,各性状均存在明显的窝效应,变化范围为0.09～0.35。胴体性状中的宰前活重、胴体重、屠宰率、瘦肉率、眼肌面积、肋膘厚、腿臀比、胴体直长、肋皮厚的遗传力分别为0.23、0.36、0.43、0.25、0.50、0.66、0.42、0.68、0.14。生长性状ADG/AGE、ADG/BF、ADG/FCR、AGE/BF、AGE/FCR、BF/FCR的遗传相关分别为-0.73、0.24、-0.51、-0.11、0.62、-0.05。胴体性状的屠宰率和瘦肉率与胴体重呈正相关关系,分别为0.43和0.17;屠宰率与瘦肉率的遗传相关为0.65,腿臀比与瘦肉率的遗传相关为0.61。说明在选育猪的肉用性状时,应考虑后腿的丰满程度。

中国大白猪SⅡ_1系选育的遗传再评定

[目的]计算中国大白猪SⅡ1系各世代种猪的估计育种值,在进行第5世代选种选配的同时,对品系选育过程进行再评定。[方法]依据湖北省农科院畜牧所中国大白猪SⅡ1系的育种资料,应用NETPIG猪育种管理软件及SAS软件进行育种值估计及数据分析。[结果]选育各世代留种公猪的综合育种指数呈上升趋势,4世代留种公猪的平均父系指数为107.44,ADG、AGE、BF、NBT、NOBA、CLR和EMA的遗传趋势分别为0.776、-0.704、-0.087、0.115、0.055、0.062和0.027。[结论]中国大白猪SⅡ1系的选育过程,在主选性状上所采取的育种措施是行之有效的,在每个世代末有必要用BLUP法对种公猪进行遗传再评定及遗传与表型趋势分析。

中国大白猪SⅡ_1系系谱结构及近交程度分析

通过整理中国大白猪SⅡ1系各世代的系谱资料,系统分析了其近交程度和世代组成的遗传结构。结果表明,目前中国大白猪SⅡ1系的平均近交系数为1.36%,仍处于较低的水平;近交系数的年增量为0.3342%,群体有效含量为149.61头。

中国大白猪SⅡ系新品系选育与利用前景

以湖北省农业科学院畜牧兽医研究所长期选育的成熟的SⅡ系大白猪为试验动物,采用现场测定的方法探讨该品系的性能潜力,评价其种用价值。SⅡ系大白猪测定群总产仔数达(11.24±2.52)头/窝,活仔数(10.40±2.42)头/窝;统计总产仔数14头/窝以上、产活仔数13头/窝以上的母猪在群体中占比分别达18.67%、18.63%,繁殖优势与潜力明显;且种猪未见繁殖障碍;环境适应能力强。以SⅡ系大白猪为基础可研发出新一代高繁殖力、高适应性、高抗逆性的母本品系,但受品系选育规模、环境制约,其良好的优势难以得到最大限度地发挥,有必要改变育种场体制,以效益促进选育进展。

中国大白猪S_(II)系繁殖性状的遗传参数估测

应用多性状动物模型DFREML方法估计了中国大白猪SII系繁殖性状的遗传参数。结果表明:总产仔数(TNB)、活产仔数(ANB)、初生窝重(LWB)、20日龄窝重(LW20)的遗传力估计值分别为0.06、0.10、0.07和0.15。20日龄窝重(LW20)的母体效应为0.20,而其它性状的母体效应均较低。各性状TNB/ANB、TNB/LWB、TNB/LW20、ANB/LWB、ANB/LW20、LWB/LW20的遗传相关分别为0.94、0.81、0.74、0.88、0.71和0.53。

电针对APP/PS1转基因小鼠海马低密度脂蛋白受体相关蛋白-1及其mRNA表达的影响

目的:研究电针是否是通过影响LRP1mRNA水平、提高LRP1的表达,从而促进脑内Aβ清除。方法:将4月龄APP/PS1转基因鼠,随机分为模型组、电针治疗组,以同窝同背景转基因阴性小鼠为正常对照组。电针干预"涌泉"、"百会"0.1 m A,15 min/次,隔日1次,治疗6周。治疗后,以免疫组化法观察脑组织LRP1阳性表达,以Western blotting法检测海马LRP1表达,以Real-time PCR法检测海马LRP1mRNA表达。结果:LRP1表达于脑微血管内皮细胞、胶质细胞、神经元等处。模型组海马LRP1蛋白、LRP1mRNA相对表达量低于正常对照组(P<0.05),电针治疗组海马LRP1及LRP1mRNA比模型组有上升趋势,但差异无统计学意义(P>0.05)。结论:电针干预可能影响5月龄APP/PS1转基因小鼠海马内所有细胞LRP1及LRP1mRNA表达,可能是电针干预AD发病的潜在靶点,但实验方案还需进一步完善。

关于B-值广义白噪声泛函的VOLTERRA型积分方程

讨论了关于Banach值广义白噪声泛函的量子Volterra型积分方程,其中核为时空白噪声,自由项为取值于B值广义泛函空间的抽象函数.在此基础上,证明了:方程解的存在性,唯一性和强连续性.

Characterization of a S-locus-related Receptor-like Kinase Cluster in Rice Chromosome 4

We have identified 14 S _locus glycoprotein (SLG)_related protein kinase genes in a 323 kb contig of rice (Oryza sativa L.) chromosome 4 and we detected the transcription pattern of this gene cluster by reverse transcription_polymerase reaction (RT_PCR). RT_PCR results revealed that nine putative genes were transcribed in rice and these genes had the different expression patterns: two genes are expressed predominantly in reproductive tissues while the other seven genes are expressed in both reproductive and vegetative tissues. Analysis of the predicted amino acid sequences demonstrated that the extracellular receptor domains are highly homologous to SLG of Brassica, whereas the cytoplasmic kinase domains contain conserved amino acids present in serine/threonine kinases.

Screening of hepatocyte proteins binding to complete S protein of hepatitis B virus by yeast-two hybrid system

AIM: To investigate the biological function of complete S protein and to look for proteins interacting with complete S protein in hepatocytes. METHODS: We constructed bait plasmid expressing complete S protein of HBV by cloning the gene of complete S protein into pGBKT7, then the recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-α-gal for selection and screening. After extracting and sequencing of plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics. RESULTS: Nineteen colonies were selected and sequenced. Among them, five colonies were Homo sapiens solute carrier family 25, member 23 (SLC25A23), one was Homo sapiens calrer.iculin, one was human serum albumin (ALB) gene, one was Homo sapiens metallothionein 2A, two were Homo sapiens betaine-homocysteine methyltransferase, three were Homo sapiens Na+ and H+coupled amino acid transport system N, one was Homo sapiens CD81 antigen (target of anti-proliferative antibody 1) (CD81), three were Homo sapiens diazepam binding inhibitor, two colonies were new genes with unknown function. CONCLUSION: The yeast-two hybrid system is an effective method for identifying hepatocyte proteins interacting with complete S protein of HBV. The complete S protein may bind to different proteins i.e., its multiple functions in vivo.

Transactivating effect of complete S protein of hepatitis B virus and cloning of genes transactivated by complete S protein using suppression subtractive hybridization technique

AIM: To investigate the transactivating effect of complete S protein of hepatitis B virus (HBV) and to construct a subtractive cDNA library of genes transactivated by complete S protein of HBV by suppression subtractive hybridization (SSH) technique and to clone genes associated with its transactivation activity, and to pave the way for elucidating the pathogenesis of hepatitis B virus infection. METHODS: pcDNA3.1(-)-complete S containing full-length HBV S gene was constructed by insertion of HBV complete S gene into BamH I/Kpn I sites. HepG2 cells were cotransfected with pcDNA3.1(-)-complete S and pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). Suppression subtractive hybridization and bioinformatics techniques were used. The mRNA of HepG2 cells transfected with pcDNA3.Incomplete S and pcDNA3.1(-) empty vector was isolated, and detected for the expression of complete S protein by reverse transcription polymerase chain reaction (RT-PCR) method, and cDNA was synthesized. After digestion with restriction enzyme RsaI, cDNA fragments were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptors 1 and 2, respectively. After tester cDNA had been hybridized with driver cDNA twice and underwent nested PCR twice, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out within E. coli strain DH5α. The cDNA was sequenced and analyzed in GenBank with BLAST search after polymerase chain reaction (PCR) amplification. RESULTS: The complete S mRNA could be detected by RT-PCR in HepG2 cells transfected with the pcDNA3.1(-)-complete S. The activity of β-gal in HepG2 cells transfected with the pcDNA3.1(-)-complete s was 6.9 times higher than that of control plasmid. The subtractive library of genes transactivated by HBV complete S protein was constructed successfully. The amplified library contains 86 positive clones. Colony PCR showed that 86 clones contained DNA inserts of 200-1 000 bp, respectively. Sequence analysis was performed in 35 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 33 coding sequences were obtained. These cDNA sequences might be target genes transactivated by complete S protein of HBV. Moreover, two unknown genes were discovered, full length coding sequences were obtained by bioinformatics techniques, one of them was named complete S transactivated protein 1 (CSTP1) and registered in GenBank (AY553877). CONCLUSION: The complete S gene of HBV has a transactivating effect on SV40 early promoter. A subtractive cDNA library of genes transactivated by HBV complete S protein using SSH technique has been constructed successfully. The obtained sequences may be target genes transactivated by HBV complete S protein among which some genes coding proteins are involved in cell cycle regulation, metabolism, immunity, signal transduction, cell apoptosis and formation mechanism of hepatic carcinoma.

PSD-95 regulates D1 dopamine receptor resensitization, but not receptor-mediated Gs-protein activation

The present study aims to define the role of postsynaptic density （PSD）-95 in the regulation of dopamine （DA） receptor function. We found that PSD-95 physically associates with either D1 or D2 DA receptors in co-transfected HEK-293 cells. Stimulation of DA receptors altered the association between D1 receptor and PSD-95 in a time-depen- dent manner. Functional assays indicated that PSD-95 co-expression did not affect DI receptor-stimulated cAMP pro- duction, Gs-protein activation or receptor desensitization. However, PSD-95 accelerated the recovery of internalized membrane receptors by promoting receptor recycling, thus resulting in enhanced resensitization of internalized D1 receptors. Our results provide a novel mechanism for regulating DA receptor recycling that may play an important role in postsynaptic DA functional modulation and synaptic neuroplasticity.

Relationship between serum S-100 protein level and ischemic damage degree in patients with acute cerebral infarction

Objective: To investigate the time course of serum S-100 concentrations of patients with acute cerebral infarction,and their relation with the clinical data and the prognosis. Methods: Serum S-100 levels were serially determined in 36 patients with acute cerebral infarction within 12 h, at 24 h and day 2, 3, 4, 5, 7 and 10 after acute cerebral infarction and in 20 age- and sex-matched control subjects. An S-100 content assay was performed using a two-site radioimmunoassay technique. The clinical status was assessed using NIH Stroke Scale. The functional deficit at 4 weeks after acute cerebral infarction was scored using the modified Rankin scale. A cranial computed tomography was performed initially. Results: Elevated concentrations of S-100 (>0.2 μg/L) were observed in 29 of 36 patients with acute cerebral infarction,but none of the control subjects. The S-100 peak levels were at day 2 and 3 after acute cerebral infarction and were significantly high in those patients with severe neurological deficit at admission, with extensive infarction or with space-occupying effect of ischemic edema as compared with the rest of the populations. Conclusion: Serum S-100 level assay can be used as a peripheral marker of ischemic brain damage, and may be helpful for evaluation of therapeutic effects in acute ischemic stroke.

Expression and purification of the complete PreS region of hepatitis B Virus

AIM: To express the complete PreS region of HBV in E.coli with good solubility and stability, and to establish an effective method for purification of the recombinant PreS protein. METHODS: The complete PreS region (PreS1 and PreS2) was fused into a series of tags including glutathione S-transferase (GST), dihydrofolate reductase (DHFR), maltose binding protein (MBP), 6× histidine, chitin binding domain (CBD), and thioredoxin, respectively. Expression of recombinant PreS fusion proteins was examined by SDS-PAGE analysis and confirmed by Western blot. Two fusion proteins, thio-PreS, and PreS-CBD, with desirable solubility and stability, were subjected to affinity purification and further characterization. RESULTS: Recombinant PreS fusion proteins could be synthesized with good yields in E.coli. However, most of these proteins except for thio-PreS and PreS-CBD were vulnerable to degradation or insoluble as revealed by SDS-PAGE and Western blot. Thio-PreS could be purified by affinity chromatography with nickel-chelating sepharose as the matrix. However, some impurities were also co-purified. A simple freeze-thaw treatment yielded most of the thio-PreS proteins in solution while the impurities were in the precipitate. Purified thio-PreS protein was capable of inhibiting the binding of HBV virion to a specific monoclonal antibody against an epitope within the PreS1 domain. CONCLUSION: Increased solubility and stability of the complete PreS region synthesized in E.coli can be achieved by fusion with the thioredoxin or the CBD tag. A simple yet highly effective method has been established for the purification of the thio-PreS protein. Purified thio-PreS protein likely assumes a native conformation, which makes it an ideal candidate for studying the structure of the PreS region as well as for screening antivirals.

Anti-angiogenesis effect of melittin on Mock/MHCC97-H cells by the regulation of cathepsin S in vivo

Objective： To study the anti-angiogenesis effect of melittin on human hepatoma Mock/MHCC97-H cells by regulatingthe expression of cathepsin S （CatS） in vivo. Methods： Models of in situ transplantation tumor of Mock/MHCC97-Hcells and silencing cathepsin shRNA-CatS/ MHCC97-H cells in nude mice were established. The model mice wererandomly divided into four groups. In the A1 group, the mice were inoculated with shRNA-CatS/MHCC97-H cells andtreated with melittin. In the A2 group, the mice were inoculated with shRNA-CatS/MHCC97-H cells and treated withsaline. In the B1 group, the mice were inoculated with Mock/MHCC97-H cells and treated with melittin. In the B2 group,the mice were inoculated with Mock/MHCC97-H cells and treated with saline. The A1 and B1 group were injected withmelittin （80 mg/kg） intraperitoneally every day. The A2 and B2 group were injected with 0.2 mL normal salineintraperitoneally every day. After administration for 25 days, the animals were sacrificed. The tumor size and weight innude mice in each group were recorded. The expression of CD34 protein in the xenograft tumor tissues was detected byimmunohistochemistry. The expression of Cat S, VEGF-A, p-VEGFR2, Ras, Raf, p-Raf, MEK1, p-MEK1, ERK1/2 andp-ERK1/2 proteins were detected by western blot. Results： The B1 group had significantly smaller tumor volumes andlower tumor weights than the B2 group （both P 〈 0.001）. There was no significant difference between the A1 group andA2 group in tumor volumes and weights. The number of CD34-positive microvessels in the B2 group was significantlyhigher than that in the A2 group （P 〈 0.001）. The number of CD34-positive microvessels in the B1 group wassignificantly lesser than that in the A1 group （P 〈 0.001）. Most strikingly, in the model featuring inoculation ofMock/MHCC97-H cells, CatS, VEGF-A, p-VEGFR2, Ras, Raf, p-Raf, MEK1, p-MEK1, ERK1/2 and p-ERK1/2expression were inhibited when treated with melittin. However, in the model featuring the inoculation ofshRNA-CatS/MHCC97-H cells, no such effects were observed with similar treatments. Conclusion： Melittin can inhibitthe growth of tumors and angiogenesis by blocking the CatS-VEGf-A signaling pathway.

EFFECT OF INTERLEUKIN-1β ON GROWTH HORMONE GENE EXPRESSION AND ITS POSSIBLE MOLECULAR MECHANISM IN RAT MtT/S SOMATOTROPH CELLS

Objective To elucidate the effect of interleukin-1β （IL- 1β） on human growth hormone （hGH） gene expression in a rat somatotropic pituitary cell line MtT/S. Methods Stably transfected MtT/S cells were firstly established by transfecting 484-Lucl plasmid which contained hGH gene promoter --484 to ＋30 bp and luciferase reporter gene. The effect of IL-1β on hGH gene expression was determined by assaying the luciferase activities. RT-PCR method was also used to determine whether IL-1 recepor mRNA was expressed in MtT/S cells. Results The 10^3 U/mL IL-1β stimulated secretion and synthesis of GH, and promoted the 5＇-promoter activity of GH gene in stably transfected MtT/SGL cells with the action of 1.38 times above the control. Among inhibitors of signaling transduction pathways, mitogen-activated protein kinase kinase （MAPKK/MEK） inhibitor PD98059 （40 μmol/L） and p38 mitogen-activated protein kinase （MAPK） inhibitor SB203580 （5 μmol/L） completely blocked the stimulatory effect of IL-1μ, and phosphatidylinositol-3-kinase （PI3-K） inhibitor LY294002 partly abolished the effect of IL-1μ. Western blot analysis further confirmed the activation of phosphorylated MEK and p38 MAPK in MtT/SGL cells. Neither over-expression of Pit- 1 nor inhibition of Pit- 1 expression affected induction of hGH promoter activity by IL-1μ. A series of deletion constructs of hGH promoter were created to identify the DNA sequence that mediated the effect of IL-1β, and results showed that the stimulatory effect of IL-1β was abolished following deletion of the --196 to -- 132 bp fragment. Conclusions IL-1β promotes GH secretion and synthesis in rat MtT/S somatotroph cells. The stimulatory effect of IL-1β on hGH gene promoter appears to require the activation of MEK, p38 MAPK, PI3-K, and a fragment of promoter sequence that spans the -196 to -132 bp of the gene, but it may be unlinked with Pit-1 protein.

S-1 induced secondary acute erythroid leukemia with a chromosome inv(12)(p13;q13)

Adjuvant chemotherapy by S-1 following gastrectomy is considered standard treatment in Japan.Analysis of follow-up data have proved the effi cacy of S-1 admin-istration,and that hematological adverse events were relatively rare.Pyrimidine anti-metabolites,including S-1,have shown relatively lower risks for secondary hematological malignancies in comparison to alkylat-ing agents and topoisomerase-Ⅱ inhibitors.We here report a case of therapy-related leukemia after S-1 administration.A patient who had received S-1as the sole adjuvant chemotherapy was diagnosed with acute erythroid leukemia.To the best of our knowledge,our patient represents the fi rst report of S-1 induced acute leukemia.

Primary malignant melanoma of the liver：One case report and literature review

Objective Primary malignant melanomas of the liver are exceedingly rare.Only 19 cases have been reported in the literature worldwide.In this report,we describe our pathological findings and review the literature in order to improve our understanding of the disease and prevent misdiagnosis,as well as provide evidence for its treatment and prognosis.Methods We present a case of an isolated malignant melanoma of the liver in a 61-year-old male Chinese patient.Results Comprehensive dermatological and ophthalmological examinations did not reveal any evidence of a primary cutaneous or ocular lesion.Similarly,serial physical examinations,auxiliary examinations,and bone scans did not demonstrate any other lesions in the brain,respiratory tract,and gastrointestinal tract.Microscopic examination of the resected specimen revealed malignant melanoma,which was confirmed by immunohistochemical staining for S-100 protein(+),ki67(30%+),EMA(+),CD10(+),and HMB-45(++).Conclusion Primary malignant melanoma may occur in the liver,and should be considered when the histopathological appearance is atypical of other hepatic neoplasms.The diagnostic criteria for hepatic malignant melanoma depend mainly on the clinical,radiographic,and histopathological findings.Pathomorphology and immumohistochemical staining can be utilized to confirm the diagnosis.