STAT3在介导雌二醇影响卵泡功能中的作用

本研究以猪卵泡为模型,探讨雌二醇(E2)经由STAT3调控生存蛋白(SURVIVIN)转录的非基因组途径,参与调控凋亡的分子机制。根据外观形态将卵泡分类,分别测定优势化卵泡与闭锁卵泡液中E2浓度。结果发现:E2在健康卵泡液中的含量极显著高于早期闭锁和晚期闭锁阶段(P<0.01)。通过荧光定量Real time-q PCR和Western Blots检测卵泡不同发育阶段颗粒细胞中STAT3/SURVIVIN m RNA和蛋白水平,结果显示:SURVIVIN和STAT3在健康卵泡的颗粒细胞中的表达量极显著高于闭锁卵泡(P<0.01),并随着闭锁程度的加剧呈现出逐渐递减的表达趋势。通过原代培养颗粒细胞中外源添加不同浓度E2(10-5、5×10-5、10-4 mol/L)检测相关基因蛋白表达水平的变化,结果显示:随着E2浓度的增加,STAT3的磷酸化水平也相应增加,其靶基因SURVIVIN的表达量却没有受到活化的STAT3影响。由此可见,E2能够通过上调STAT3的表达参与调控卵泡闭锁的发生。

Genome-Wide Association Study for Certain Carcass Traits and Organ Weights in a Large White×Minzhu Intercross Porcine Population

Porcine carcass traits and organ weights have important economic roles in the swine industry. A total of 576 animals from a Large White×Minzhu intercross population were genotyped using the Illumina PorcineSNP60K Beadchip and were phenotyped for 10 traits, speciifcally, backfat thickness （6-7 libs）, carcass length, carcass weight, foot weight, head weight, heart weight, leaf fat weight, liver weight, lung weight and slaughter body weight. The genome-wide association study （GWAS） was assessed by Genome Wide Rapid Association using the mixed model and regression-genomic control approach. A total of 31 single nucleotide polymorphisms （SNPs） （with the most signiifcant SNP being MARC0033464, P value=6.80×10-13） were located in a 9.76-Mb （31.24-41.00 Mb） region on SSC7 and were found to be signiifcantly associated with one or more carcass traits and organ weights. High percentage of phenotypic variance explanation was observed for each trait ranging from 31.21 to 67.42%. Linkage analysis revealed one haplotype block of 495 kb, in which the most signiifcant SNP being MARC0033464 was contained, on SSC7 at complete linkage disequilibrium. Annotation of the pig reference genome suggested 6 genes （GRM4, HMGA1, NUDT3, RPS10, SPDEF and PACSIN1） in this candidate linkage disequilibrium （LD） interval. Functional analysis indicated that the HMGA1 gene presents the prime biological candidate for carcass traits and organ weights in pig, with potential application in breeding programs.

Heritabilities and genetic and phenotypic correlations of litter uniformity and litter size in Large White sows

Litter uniformity, which is usually represented by within-litter weight coefficient of variation at birth （CVB）, could influence litter performance of sows and the profitability of pig enterprises. The objective of this study was to characterize CVB and its effect on other reproductive traits in Large White sows. Genetic parameters and genetic correlation of the reproductive traits, including CVB, within-litter weight coefficient of variation at three weeks （CVT）, total number born （TNB）, number born alive （NBA）, number born dead （NBD）, gestation length （GL）, piglet mortality at birth （Mo）, piglet mortality at three weeks （M3）, total litter weight at birth （TLW0）, and total litter weight at three weeks （TLW3） were estimated for 2 032 Large White litters. The effects of parity and classified litter size on CVB, CVT, TNB, NBA, NBD, GL, M0, M3, TLW0, and TLW3 were also estimated. The heritabilities of these reproductive traits ranged from 0.06 to 0.17, with the lowest heritability for CVB and the highest heritability for TLW0. Phenotypic and genetic correlations between these reproductive traits were low to highly positive and negative （ranging from -0.03 to 0.93, and -0.53 to 0.93, respectively）. The genetic correlations between TNB and CVB, and between M0 and CVB were 0.32 and 0.29, respectively. In addition, CVB was significantly influenced by parity and litter size class （P〈0.05）. All the results suggest that piglet uniformity should be maintained in pig production practices and pig breeding programs.

Quantitative trait loci for the number of vertebrae on Sus scrofa chromosomes 1 and 7 independently influence the numbers of thoracic and lumbar vertebrae in pigs

Although quantitative trait loci（QTLs） for number of thoracic-lumbar vertebrae have been identified on Sus scrofa chromosomes（SSCs） 1 and 7, the influence of these QTLs on the thoracic and lumbar vertebrae is not clear. The aim of this study was to identify single nucleotide polymorphisms（SNPs） associated with total number of thoracic-lumbar vertebrae and for each trait（number of thoracic and lumbar vertebrae） separately. A total of 581 individuals from an F2 Large White×Minzhu population were genotyped using an SNP60 K chip. Performing a genome-wide association study（GWAS） for total number of thoracic-lumbar vertebrae, 38 significant SNPs were identified in two QTL regions located on SSC1 and SSC7. Performing a GWAS for number of thoracic vertebrae only, 72 significant SNPs were located on SSC7. While performing a GWAS for number of lumbar vertebrae only, 17 significant SNPs were identified on SSC1. Gene mining suggested that the gene encoding orphan nuclear receptor, germ cell nuclear factor（NR6A1） on SSC1 was a strong candidate affecting the number of lumbar vertebrae in pigs. Additionally, genes encoding vertnin（VRTN）, prospero homeobox 2（PROX2）, Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog（FOS）, and transforming growth factor beta 3（TGFB3） may be important candidates affecting the number of thoracic vertebrae in pigs. QTLs on SSC1 and SSC7 independently influenced the numbers of thoracic and lumbar vertebrae. These results shed light on the complex genetic background of vertebrae development in pigs.

一种新型二氢吡喃并[2,3-c]吡唑类衍生物的体外抑瘤作用及机制

目的研究一种新型二氢吡喃并[2,3-c]吡唑类衍生物(36号化合物)的体外抑瘤作用及可能的分子机制。方法 MTT比色法检测36号化合物对人乳腺癌细胞Bcap-37,MCF-7和人肺癌细胞A549体外存活的影响,反转录荧光定量PCR法检测Bcap-37细胞凋亡相关基因p53,p21,Bax和NF-κB mRNA水平,流式细胞术检测Bcap-37细胞周期和细胞凋亡,JC-1荧光探针法测定Bcap-37细胞线粒体膜电位(MMP)。结果 36号化合物对Bcap-37,MCF-7和A549细胞存活均表现出浓度依赖性抑制作用,其中对人乳腺癌细胞Bcap-37的半数抑制浓度(IC50)最低,为(22.4±0.8)mg·L^(-1)。与正常对照组相比,36号化合物在25 mg·L^(-1)浓度下使Bcap-37细胞周期阻滞于G1期(P<0.01),早期凋亡细胞显著增加(P<0.01);在50 mg·L^(-1)浓度下,使Bcap-37细胞凋亡相关基因p53,p21,Bax和NF-κB mRNA水平均显著升高(P<0.05,P<0.01);在10,25和75 mg·L^(-1)浓度下,使Bcap-37细胞MMP明显降低(P<0.05,P<0.01)。结论 36号化合物具有良好的体外抑瘤作用,可抑制Bcap-37细胞增殖,促进其凋亡,其作用机制与细胞内线粒体通路和细胞外凋亡信号通路的激活相关。

Genome-wide association study for rib eye muscle area in a Large White×Minzhu F_2 pig resource population

Rib eye muscle area（REMA） is an economically important trait and one of the main selection criteria for breeding in the swine industry. In the genome-wide association study（GWAS）, the Illumina Porcine SNP60 Bead Chip containing 62 163 single nucleotide polymorphisms（SNPs） was used to genotype 557 pigs from a porcine Large White×Minzhu intercross population. The REMA（at the 5th–6th, 10th–11th and the last ribs） was measured after slaughtered at the age of（240±7） d for each animal. Association tests between REMA trait and SNPs were performed via the Genome-Wide Rapid Association using the Mixed Model and Regression-Genomic Control（GRAMMAR-GC） approach. From the Ensembl porcine database, SNP annotation was implemented using Sus scrofa Build 10.2. Thirty-three SNPs on SSC12 and 3 SNPs on SSC2 showed significant association with REMA at the last rib at the chromosome-wide significance level. None of the SNPs of REMA at the 5th–6th rib and only a few numbers of the SNPs of REMA at the 10th–11th ribs were found in this study. The Haploview V3.31 program and the Haplo.Stats R package were used to detect and visualize haplotype blocks and to analyze the association of the detected haplotype blocks with REMA at the last rib. A linkage analysis revealed that 4 haplotype blocks contained 4, 4, 2, and 4 SNPs, respectively. Annotations from pig reference genome suggested 2 genes（NOS2, NLK） in block 1（266 kb）, one gene（TMIGD1） in block 2（348 kb）, and one gene（MAP2K4） in block 3（453 kb）. A functional analysis indicated that MYH3 and MYH13 genes are the potential genes controlling REMA at the last rib. We screened several candidate intervals and genes based on the SNPs location and the gene function, and inferred that NOS2 and NLK genes maybe the main genes of REMA at the last ribs.

聚苯胺/聚吡咯共聚物的制备及其对磷酸根吸附性能研究

通过改变苯胺和吡咯的比例,用循环伏安法制得了聚苯胺/聚吡咯(PANi/PPy)共聚物。采用拉曼光谱、扫描电镜表征了其结构特征,并研究了其对磷酸根的吸附性能。结果表明:吡咯的加入提高了苯胺聚合速率,改变了聚合物结构,团聚现象受到抑制,产物趋向于形成松散且有一定规则的片状结构。吸附研究表明:PANi/PPy对水中磷酸根具有较好的吸附效果,Langmuir方程和准二级动力学方程很好地说明了吸附过程。

Porcine LEM domain-containing 3: Molecular cloning, functional characterization, and polymorphism associated with ear size

Ear size exhibits remarkable diversity in pig breeds. LEM domain-containing 3 （LEMD3） on chromosome 5 is considered as an important candidate for porcine ear size. This is the first study on cloning and characterization of LEMD3 cDNA. The complete cDNA contains 4 843 bp, including a 2 736-bp open reading frame （ORF）, a 37-bp 5＂-untranslated region （UTR） and a 2070-bp 3＂-UTR. The complete LEMD3 gene is 126241-bp and contains 13 exons and 12 introns. The ORF encodes a deduced LEMD3 protein of 911 amino acids, which shares 82-94% nucleic acid and 51-96% amino acid identity with other species. A phylogenetic tree constructed based on the amino acid sequences revealed that the porcine LEMD3 protein was closely related with cattle LEMD3. Resequencing of the ORF and promoter of LEMD3 from Minzhu pig and Large White revealed three single nucleotide polymorphisms （SNPs）： L964C〉A in the complete coding region, L4625A〉G in the 3＂ UTR, and L-394T〉C in the promoter region. Genome-wide association study （GWAS） revealed that all of SNPs were shown significant association with ear size in Large WhitexMinzhu pig intercross population. With conditional GWAS, -Iogl0（P-value） decreased by more than 80% when each of three SNPs was included as a fixed effect. These results suggested direct involvement of LEMD3 or close linkage to the causative mutation for ear size. The findings of this study might form the basis for understanding the genetic mechanism of ear size variation in pigs and provide potential molecular markers for screening ear size diversity in pig breeds.