masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD...masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD和bamA的实时荧光定量PCR引物,将质粒DNA进行8次10倍梯度稀释后构建实时荧光定量PCR标准曲线.优化后的体系(20μL)为:FastStart Essential DNA Green Master 10.0μL,上下游引物各0.4μL,RNase-Free Water 4.2μL,5.0μL DNA模板.利用新设计的引物扩增masD和bamA基因的最适退火温度分别为61℃和57℃.优化后的检测方法扩增效率提高至97.5%和71.2%,比文献报道的方法提高了7.6%—44.5%,具有更高的重复性和灵敏度.利用设计的引物对陕北5个地区石油污染土壤中的masD和bamA基因进行定量检测结果表明,石油污染土壤中普遍存在着控制石油烃厌氧降解的关键基因,所测定的土壤中bamA降解基因的拷贝数远高于masD降解基因.展开更多
Background:Hundreds of single-nucleotide polymorphism(SNP)sites have been found to be potential genetic markers of type 2 diabetes mellitus(T2DM).However,SNPs related to T2DM in minipigs have been less reported.This s...Background:Hundreds of single-nucleotide polymorphism(SNP)sites have been found to be potential genetic markers of type 2 diabetes mellitus(T2DM).However,SNPs related to T2DM in minipigs have been less reported.This study aimed to screen the T2DM-susceptible candidate SNP loci in Bama minipigs so as to improve the success rate of the minipig T2DM model.Methods:The genomic DNAs of three Bama minipigs with T2DM,six sibling lowsusceptibility minipigs with T2DM,and three normal control minipigs were compared by whole-genome sequencing.The T2DM Bama minipig-specific loci were obtained,and their functions were annotated.Meanwhile,the Biomart software was used to perform homology alignment with T2DM-related loci obtained from the human genome-wide association study to screen candidate SNP markers for T2DM in Bama miniature pigs.Results:Whole-genome resequencing detected 6960 specific loci in the minipigs with T2DM,and 13 loci corresponding to 9 diabetes-related genes were selected.Further,a set of 122 specific loci in 69 orthologous genes of human T2DM candidate genes were obtained in the pigs.Collectively,a batch of T2DM-susceptible candidate SNP markers in Bama minipigs,covering 16 genes and 135 loci,was established.Conclusions:Whole-genome sequencing and comparative genomics analysis of the orthologous genes in pigs that corresponded to the human T2DM-related variant loci successfully screened out T2DM-susceptible candidate markers in Bama miniature pigs.Using these loci to predict the susceptibility of the pigs before constructing an animal model of T2DM may help to establish an ideal animal model.展开更多
After analysis of characteristics of Bama rural health-caring tourism,it had highlighted monopolisticness of longevity culture,diversity of tourist resources,and harmony between human and nature.The paper pointed out ...After analysis of characteristics of Bama rural health-caring tourism,it had highlighted monopolisticness of longevity culture,diversity of tourist resources,and harmony between human and nature.The paper pointed out that,to accelerate new countryside construction of Bama rural health-caring tourism,it should realize nearby employment and increase farmers' income;complete construction of rural basic facilities;ameliorate rural sanitary condition and transform villagers' thought;promote adjustment of rural industrial structure;and establish protection mechanism of longevous people.After survey on development of Bama rural tourism,it concluded bottlenecks restricting Bama rural tourism development,which were that environment at longevity area had suffered pollution;that the development was disordered and lacked of management;and that basic facilities waited for improvement.On this basis,approaches for improvement of competitiveness of Bama health-caring tourism had been proposed.It emphasized that it should enhance protection of longevity tourist resources,build farmhouse inn demonstration base,implement admittance system,establish community health-care system,beneficial to realizing economic,social and ecological benefits,and further manifesting its exuberant vitality.展开更多
[ Objective] To clone and analyze the sequence of Adiponectin receptor 1 （ AdipoR1 ） and receptor 2 （AdipoR2） cDNA of Guangxi Bama mini-pig. [Method] The Adiponectin receptors cDNAs were amplified by RT-PCR using ...[ Objective] To clone and analyze the sequence of Adiponectin receptor 1 （ AdipoR1 ） and receptor 2 （AdipoR2） cDNA of Guangxi Bama mini-pig. [Method] The Adiponectin receptors cDNAs were amplified by RT-PCR using skeletal muscle total RNA as template and then ligated into pMD18-T vector after purification. The recombinant pMD18-T vector was transformed into the E. coil DH5α for identification and sequencing. And the results were compared with the cDNA sequence from other species. [Result] The fragments, 1 128 bp and 1 161 bp in size, were amplified by RT-PCR and respectively consistent with the coding sequence of AdipoR1 gene and AdipoR2 gene. The homology analysis showed that the sequences of AdipoR1 gene and AdipoR2 gene were respectively 99.8% and 99.7% homologous to the sequence of domestic pig reported in GenBank with one base and three base missense mutations correspondingly. [ Conclusion] The AdipoR1 gene and AdipoR2. gene were successfully amplified from Guangxi Bama mini-pig, laying the foundation for the further study of the biological function of AdipoR genes and the design of novel drugs with AdipoR as target.展开更多
masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物ma...masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物masD-f、masD-r和bamA-f、bamA-r。经过常规PCR扩增分别得到片段大小为389和354 bp扩增产物,经测序并在NCBI数据库查询,确定为masD和bamA片段。通过实时荧光定量PCR构建测定这两种基因的标准曲线。优化后的扩增体系(25μL)为:12.5μL 2×Trans Start Top Green qPCR Super Mix,引物浓度为0. 2μmol/L,masD和bamA基因最适退火温度分别为52℃和56℃。建立的两种基因的实时荧光定量PCR检测方法具有非常好的重复性,其灵敏度比传统PCR技术高100倍。对于氧化石墨烯促进石油烃的厌氧降解体系中厌氧基因的定量检测显示,添加不同浓度的石墨烯均促进了bamA拷贝数的增加,但对masD的拷贝数无显著影响。展开更多
The Bama Xiang pig （BMX） Chinese indigenous breed is a famous early-maturing with a two-end black coat To uncover the genetic basis of the BMX phenotype, we conducted comparative genomic analyses between BMX and Eas...The Bama Xiang pig （BMX） Chinese indigenous breed is a famous early-maturing with a two-end black coat To uncover the genetic basis of the BMX phenotype, we conducted comparative genomic analyses between BMX and East Asian wild boars and Laiwu pigs, respectively. Genes under positive selection were enriched in pathways associated with gonadal hormone and melanin synthesis, consistent with the phenotypic changes observed during development in BMX pigs. We also performed differentially expressed gene analysis based on RNA-seq data from pituitary tissues of BMX and Large White pigs. The CTTNBP2NL, FRS2, KANK4, and KATNAL1 genes were under selection and exhibited expressional changes in the pituitary tissue, which may affect BMX pig puberty. Our study demonstrated the positive selection of early maturity in the development of BMX pigs and advances our knowledge on the role of regulatory elements in puberty evolution in pigs.展开更多
Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity ofgene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major ...Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity ofgene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlation between alternative splicing and heat stress in pigs. Therefore, this study aimed to clone, sequence and quantify the alternative splicing variant of toll-like receptor 4 （TLR4） in Bama miniature pigs （Sus scrofa domestica） following exposure to heat stress. The results showed that the second exon of TLR4 was spliced and 167 bp shorter in the alternative splicing variant, and the protein was putatively identified as a type of truncated membrane protein consisting of extramembrane, transmembrane and intramembrane regions lacking a signal peptide. Further, it was not a non- classical secretory protein. Five potential reference genes were screened for their potential as reliable standards to quantify the expression of TLR4 alternative spliced variants by real-time quantitative reverse-transcriptase polymerase chain reaction （qRT-PCR）. The stability of these reference genes was ranked using the geNorm and NormFinder programs, and ribosomal protein L4 （RPL4） and TATA box-binding protein （TBP） were found to be the two genes showing the most stable expression in the in vitro cultured peripheral blood mononuclear ceils （PBMCs） during heat shock. The mRNA level of the TLR4 gene （both classical and spliced） in stressed pigs increased significantly （P〈0.05）. Further, the expression levels of the alternative spliced variant of TLR4 （TLR4-ASV） showed a 2-3 folds increase in heat-stressed PBMCs as compared to control pigs. The results of the present study suggested that heat shock might modulate the host immune response by regulating the expressions of TLR4 and its alternative splicing variant.展开更多
Summary： Somatic cell nucleus transfer （SCNT） has been considered the most effective method for conserving endangered animals and expanding the quantity of adult animal models. Bama miniature pigs are genetically s...Summary： Somatic cell nucleus transfer （SCNT） has been considered the most effective method for conserving endangered animals and expanding the quantity of adult animal models. Bama miniature pigs are genetically stable and share similar biological features to humans. These pigs have been used to establish animal models for human diseases, and for many other applications. However, there is a pan- city of studies on the effect of ear fibroblasts derived from different age of adult Bama miniature pigs on nucleus transfer （NT）. The present study examined the NT efficiency of ear fibroblasts from fetal, new- born, 1-, 2-, 4-, 6-, 12-month-old miniature pigs by using trypan blue staining, flow cytometry and NT technique, etc., and the cell biological function and SCNT efficiency were compared between groups. The results showed that ear fibroblasts grew well after passage in each group. Spindle-shaped cells ini- tially predominated, and gradually declined with increase of culture time and replaced by polygonal cells. Irregular cell growth occurred in the 2-month-old group and the elder groups. The growth curves of the ear fibroblasts were ＂S-shaped＂ in different age groups. The cell proliferation of postnatal ear fi- broblasts, especially those from 2-, 4-, 6-, 12-month-old miniature pigs was significantly different from that of fetus ear fibroblasts （P〈0.05 or P〈0.01）. Two-month- and 4-month-old ear fibroblasts had a sig- nificantly higher proportion of G1 stage cells （85% to 91%） than those at 6 and 12 months （66% to 74%, P〈0.01）. The blastocyst rate of reconstructed embryos originating from newborn, 1-, 2-, 4-month-old donor pigs was 6.06% to 7.69% with no significant difference from that in fetus fibroblast group （8.06%）. It was concluded that 〈4-month-old adult Bama miniature pigs represent a better donor cell resource than elder pigs.展开更多
[ Objective] The research aimed to discuss the relationship between the polymorphism of PPARy.2 gene and the susceptibility of type 2 diabetes mellitus （T2DM） in Guangxi Bama mini-pigs. [ Method] 24 Guangxi Bama min...[ Objective] The research aimed to discuss the relationship between the polymorphism of PPARy.2 gene and the susceptibility of type 2 diabetes mellitus （T2DM） in Guangxi Bama mini-pigs. [ Method] 24 Guangxi Bama mini-pigs were fed with high-fat and high-sucrose diet, and partial sequences of exon 2 of PPARy-2 gene were amplified by using PCR method. In addition, the contents of fasting blood glucose and insulin （INS） in Guangxi Bama mini-pigs were determined, and the glucose tolerance test （GTT） was also carried out. [ Result] There was one SNP site （19813A/G） Jn partial sequence of exon 2 of the cloned PPAFly-2 gene, and AA （7 pigs） and AG （17 pigs） genotype were detected. The contents of fasting insulin and 60-min blood glucose in GTT in AG-genotype Guangxi Bama mini-pigs were significantly higher than those of AA genotype （ P 〈0.05）, while the incidence of T2DM in AG-genotype Guangxi Bama mini-pigs （71.4%） was obviously higher than that of AA gen- otype （5.9%）. [ Conclusion] The polymorphism of 19813A/G in exon 2 of PPARy-2 gene was related with the susceptibility of T2DM in Guangxi Bama mini-pigs.展开更多
Bama County。in the Guangxi Zhuang Autonomous Region in South China.is well known for a longer meanage of its population in China and the world over.It is called a 1and of longevity,and there are at present 66 centena...Bama County。in the Guangxi Zhuang Autonomous Region in South China.is well known for a longer meanage of its population in China and the world over.It is called a 1and of longevity,and there are at present 66 centenarians in the county.展开更多
Objective: To study the interaction between the polymorphisms of FOXO3A and the environmental factors in the longevity district of Bama county.Methods: Using Taqmaq probe,genotyping of FOXO3A gene was performed for 20...Objective: To study the interaction between the polymorphisms of FOXO3A and the environmental factors in the longevity district of Bama county.Methods: Using Taqmaq probe,genotyping of FOXO3A gene was performed for 200 longevous elderly(aged 90-110)and 288 healthy controls(aged 48-89).The interaction between the polymorphisms of gene and environmental factors were analyzed by using conditional logistic regression.Results: Minor allele frequencies(MAF)of five known SNPs in FOXO3A gene were higher in cases than in controls(P<0.05).There was a positive interction between minority alleles of rs2764264 in FOXO3A gene and the environment factors in the longevity district(OR=11.08,γ=1.075).Conclusion: Five SNPs of FOXO3A gene would be closely correlated with human longevity in Bama.The interaction between the polymorphisms of FOXO3A genes rs2764264 and the environment factors in the longevity district had synergy to longevity.The longevity phenomenon in Bama probably was the result of the interaction between gene and environmental factors.展开更多
The gastrointestinal tract is essential for food digestion,nutrient absorption,waste elimination,and microbial defense.Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding...The gastrointestinal tract is essential for food digestion,nutrient absorption,waste elimination,and microbial defense.Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding of cellular diversity,functional heterogeneity,and their importance in intestinal tract development and disease.Although such profiling has been extensively conducted in humans and mice,the single-cell gene expression landscape of the pig cecum remains unexplored.Here,single-cell RNA sequencing was performed on 45572 cells obtained from seven cecal samples in pigs at four different developmental stages(days(D)30,42,150,and 730).Analysis revealed 12 major cell types and 38 subtypes,as well as their distinctive genes,transcription factors,and regulons,many of which were conserved in humans.An increase in the relative proportions of CD8^(+)T and Granzyme A(low expression)natural killer T cells(GZMA^(low)NKT)cells and a decrease in the relative proportions of epithelial stem cells,Tregs,RHEX^(+)T cells,and plasmacytoid dendritic cells(pDCs)were noted across the developmental stages.Moreover,the post-weaning period exhibited an up-regulation in mitochondrial genes,COX2 and ND2,as well as genes involved in immune activation in multiple cell types.Cell-cell crosstalk analysis indicated that IBP6^(+)fibroblasts were the main signal senders at D30,whereas IBP6^(−)fibroblasts assumed this role at the other stages.NKT cells established interactions with epithelial cells and IBP6^(+)fibroblasts in the D730 cecum through mediation of GZMA-F2RL1/F2RL2 pairs.This study provides valuable insights into cellular heterogeneity and function in the pig cecum at different development stages.展开更多
Introduction: Pesticides are currently an essential component of agricultural production techniques for controlling pests and weeds. In Burkina Faso, non-compliance with good practice in the use of pesticides poses a ...Introduction: Pesticides are currently an essential component of agricultural production techniques for controlling pests and weeds. In Burkina Faso, non-compliance with good practice in the use of pesticides poses a real health problem for the population. This study examines the health risks associated with pesticide management in rice-growing areas. Material and Methods: A field survey was conducted in Bama, involving farmers, focusing on their socio-demographic characteristics, pesticide usage, and health effects. Cholinesterase levels were measured in subsample of farmers using a portable device. Data were analysed using Microsoft Excel, calculating means and percentages for various practices. Health consultations, protection methods, and pesticide management were studied. Erythrocyte acetylcholinesterase activity was compared before and after treatment. Data were categorised into classes based on inhibition levels, and correlation analyses determined relationships between variables such as age, years of experience, and cholinesterase activity. Results: The results indicate that rice cultivation is mainly carried out by a fairly young population, with nearly 63% being under the age of 50. Common poor practices in pesticide use include improper storage and reuse of leftover pesticides. Seven types of pesticides were identified, including organophosphates such as glyphosate, which was used in 26.7% of cases. This organophosphate has resulted in class B poisoning, causing a 30% - 50% reduction in erythrocyte cholinesterase activity. The health effects of pesticide use are felt by agricultural farmers through various symptoms of poisoning. Conclusion: To reduce the occurrence of pesticide poisoning, it is essential to launch information and awareness campaigns among the population and farmers to promote safe practices in pesticide use in Bama, Burkina Faso.展开更多
文摘目的以革兰阴性菌外膜蛋白折叠辅助因子关键蛋白BamA为靶蛋白,基于生物膜干涉(Biolayer interferometry,BLI)技术建立化合物与BamA蛋白β折叠结构域(BamA_(β-barrel))结合活性的评价方法,为建立靶向BamA蛋白的抗革兰阴性菌先导物奠定基础。方法应用BLI方法检测BamA_(β-barrel)与已知的阳性化合物darobactin的结合活性。原核表达并纯化带有His标签的大肠埃希菌BamA_(β-barrel)蛋白,使用表面活性剂LDAO对其进行复性和折叠;使用生物素标记折叠和未折叠蛋白,并结合到超级链霉亲和素(super streptavidin,SSA)生物传感器,然后检测蛋白与不同浓度的darobactin结合信号的变化,同时做无蛋白或darobactin稀释液对照;空白对照采用未结合生物素化的BamA_(β-barrel)蛋白的传感器,检测上述系列稀释样品。相应信号采用Steady state analysis方式拟合分析,计算平衡常数(KD)值。结果成功获得高纯度的折叠状态BamA_(β-barrel)蛋白,通过BLI技术检测到折叠状态的BamA_(β-barrel)与阳性化合物darobactin具有良好结合活性且呈现浓度依赖性,R^(2)为0.9998,KD值为(2.2E-06±8.0E-08)M。结论基于BLI技术成功建立了折叠状态的BamA_(β-barrel)-化合物结合活性的评价方法,为后续BamA蛋白靶向性抗革兰阴性菌抗生素的发现建立基础。
文摘masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD和bamA的实时荧光定量PCR引物,将质粒DNA进行8次10倍梯度稀释后构建实时荧光定量PCR标准曲线.优化后的体系(20μL)为:FastStart Essential DNA Green Master 10.0μL,上下游引物各0.4μL,RNase-Free Water 4.2μL,5.0μL DNA模板.利用新设计的引物扩增masD和bamA基因的最适退火温度分别为61℃和57℃.优化后的检测方法扩增效率提高至97.5%和71.2%,比文献报道的方法提高了7.6%—44.5%,具有更高的重复性和灵敏度.利用设计的引物对陕北5个地区石油污染土壤中的masD和bamA基因进行定量检测结果表明,石油污染土壤中普遍存在着控制石油烃厌氧降解的关键基因,所测定的土壤中bamA降解基因的拷贝数远高于masD降解基因.
基金National Natural Science Foundation of China,Grant/Award Number:3147205731802021.Specialized Research Fund for Laboratory Animal Science of PLA,Grant/Award Number:SYDW[2020]01SYDW[2020]02.
文摘Background:Hundreds of single-nucleotide polymorphism(SNP)sites have been found to be potential genetic markers of type 2 diabetes mellitus(T2DM).However,SNPs related to T2DM in minipigs have been less reported.This study aimed to screen the T2DM-susceptible candidate SNP loci in Bama minipigs so as to improve the success rate of the minipig T2DM model.Methods:The genomic DNAs of three Bama minipigs with T2DM,six sibling lowsusceptibility minipigs with T2DM,and three normal control minipigs were compared by whole-genome sequencing.The T2DM Bama minipig-specific loci were obtained,and their functions were annotated.Meanwhile,the Biomart software was used to perform homology alignment with T2DM-related loci obtained from the human genome-wide association study to screen candidate SNP markers for T2DM in Bama miniature pigs.Results:Whole-genome resequencing detected 6960 specific loci in the minipigs with T2DM,and 13 loci corresponding to 9 diabetes-related genes were selected.Further,a set of 122 specific loci in 69 orthologous genes of human T2DM candidate genes were obtained in the pigs.Collectively,a batch of T2DM-susceptible candidate SNP markers in Bama minipigs,covering 16 genes and 135 loci,was established.Conclusions:Whole-genome sequencing and comparative genomics analysis of the orthologous genes in pigs that corresponded to the human T2DM-related variant loci successfully screened out T2DM-susceptible candidate markers in Bama miniature pigs.Using these loci to predict the susceptibility of the pigs before constructing an animal model of T2DM may help to establish an ideal animal model.
文摘After analysis of characteristics of Bama rural health-caring tourism,it had highlighted monopolisticness of longevity culture,diversity of tourist resources,and harmony between human and nature.The paper pointed out that,to accelerate new countryside construction of Bama rural health-caring tourism,it should realize nearby employment and increase farmers' income;complete construction of rural basic facilities;ameliorate rural sanitary condition and transform villagers' thought;promote adjustment of rural industrial structure;and establish protection mechanism of longevous people.After survey on development of Bama rural tourism,it concluded bottlenecks restricting Bama rural tourism development,which were that environment at longevity area had suffered pollution;that the development was disordered and lacked of management;and that basic facilities waited for improvement.On this basis,approaches for improvement of competitiveness of Bama health-caring tourism had been proposed.It emphasized that it should enhance protection of longevity tourist resources,build farmhouse inn demonstration base,implement admittance system,establish community health-care system,beneficial to realizing economic,social and ecological benefits,and further manifesting its exuberant vitality.
基金Supported by Guangxi Science Foundation (0542025)~~
文摘[ Objective] To clone and analyze the sequence of Adiponectin receptor 1 （ AdipoR1 ） and receptor 2 （AdipoR2） cDNA of Guangxi Bama mini-pig. [Method] The Adiponectin receptors cDNAs were amplified by RT-PCR using skeletal muscle total RNA as template and then ligated into pMD18-T vector after purification. The recombinant pMD18-T vector was transformed into the E. coil DH5α for identification and sequencing. And the results were compared with the cDNA sequence from other species. [Result] The fragments, 1 128 bp and 1 161 bp in size, were amplified by RT-PCR and respectively consistent with the coding sequence of AdipoR1 gene and AdipoR2 gene. The homology analysis showed that the sequences of AdipoR1 gene and AdipoR2 gene were respectively 99.8% and 99.7% homologous to the sequence of domestic pig reported in GenBank with one base and three base missense mutations correspondingly. [ Conclusion] The AdipoR1 gene and AdipoR2. gene were successfully amplified from Guangxi Bama mini-pig, laying the foundation for the further study of the biological function of AdipoR genes and the design of novel drugs with AdipoR as target.
文摘masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物masD-f、masD-r和bamA-f、bamA-r。经过常规PCR扩增分别得到片段大小为389和354 bp扩增产物,经测序并在NCBI数据库查询,确定为masD和bamA片段。通过实时荧光定量PCR构建测定这两种基因的标准曲线。优化后的扩增体系(25μL)为:12.5μL 2×Trans Start Top Green qPCR Super Mix,引物浓度为0. 2μmol/L,masD和bamA基因最适退火温度分别为52℃和56℃。建立的两种基因的实时荧光定量PCR检测方法具有非常好的重复性,其灵敏度比传统PCR技术高100倍。对于氧化石墨烯促进石油烃的厌氧降解体系中厌氧基因的定量检测显示,添加不同浓度的石墨烯均促进了bamA拷贝数的增加,但对masD的拷贝数无显著影响。
基金supported by grants from the National Natural Science Foundation of China(31472000)National 973 Program of China(2013CB835203)Animal Branch of the Germplasm Bank of Wild Species,Chinese Academy of Sciences(Large Research Infrastructure Funding)
文摘The Bama Xiang pig （BMX） Chinese indigenous breed is a famous early-maturing with a two-end black coat To uncover the genetic basis of the BMX phenotype, we conducted comparative genomic analyses between BMX and East Asian wild boars and Laiwu pigs, respectively. Genes under positive selection were enriched in pathways associated with gonadal hormone and melanin synthesis, consistent with the phenotypic changes observed during development in BMX pigs. We also performed differentially expressed gene analysis based on RNA-seq data from pituitary tissues of BMX and Large White pigs. The CTTNBP2NL, FRS2, KANK4, and KATNAL1 genes were under selection and exhibited expressional changes in the pituitary tissue, which may affect BMX pig puberty. Our study demonstrated the positive selection of early maturity in the development of BMX pigs and advances our knowledge on the role of regulatory elements in puberty evolution in pigs.
基金supported by grants from the National Natural Science Foundation of China (31101862)the China Postdoctor Science Foundation and Guangdong Ocean University Doctor Seed Grant, China (0712107)
文摘Alternative splicing is a cellular mechanism in eukaryotes that results in considerable diversity ofgene products. It plays an important role in several diseases and cellular signal regulation. Heat stress is a major factor that induces immunosuppression in pigs. Little is known about the correlation between alternative splicing and heat stress in pigs. Therefore, this study aimed to clone, sequence and quantify the alternative splicing variant of toll-like receptor 4 （TLR4） in Bama miniature pigs （Sus scrofa domestica） following exposure to heat stress. The results showed that the second exon of TLR4 was spliced and 167 bp shorter in the alternative splicing variant, and the protein was putatively identified as a type of truncated membrane protein consisting of extramembrane, transmembrane and intramembrane regions lacking a signal peptide. Further, it was not a non- classical secretory protein. Five potential reference genes were screened for their potential as reliable standards to quantify the expression of TLR4 alternative spliced variants by real-time quantitative reverse-transcriptase polymerase chain reaction （qRT-PCR）. The stability of these reference genes was ranked using the geNorm and NormFinder programs, and ribosomal protein L4 （RPL4） and TATA box-binding protein （TBP） were found to be the two genes showing the most stable expression in the in vitro cultured peripheral blood mononuclear ceils （PBMCs） during heat shock. The mRNA level of the TLR4 gene （both classical and spliced） in stressed pigs increased significantly （P〈0.05）. Further, the expression levels of the alternative spliced variant of TLR4 （TLR4-ASV） showed a 2-3 folds increase in heat-stressed PBMCs as compared to control pigs. The results of the present study suggested that heat shock might modulate the host immune response by regulating the expressions of TLR4 and its alternative splicing variant.
基金supported by the grants from the National Natural Science Foundation of China(No.31000546)National High-tech Research & Development Program of China(863 Program)(No.2012AA020603)+1 种基金National Science and Technology Major Project of China(No.2014zx08009-003-006)Rongchang Youth Foundation and Fundamental Research Funds of Southwest University(No.XDJK2012C097)
文摘Summary： Somatic cell nucleus transfer （SCNT） has been considered the most effective method for conserving endangered animals and expanding the quantity of adult animal models. Bama miniature pigs are genetically stable and share similar biological features to humans. These pigs have been used to establish animal models for human diseases, and for many other applications. However, there is a pan- city of studies on the effect of ear fibroblasts derived from different age of adult Bama miniature pigs on nucleus transfer （NT）. The present study examined the NT efficiency of ear fibroblasts from fetal, new- born, 1-, 2-, 4-, 6-, 12-month-old miniature pigs by using trypan blue staining, flow cytometry and NT technique, etc., and the cell biological function and SCNT efficiency were compared between groups. The results showed that ear fibroblasts grew well after passage in each group. Spindle-shaped cells ini- tially predominated, and gradually declined with increase of culture time and replaced by polygonal cells. Irregular cell growth occurred in the 2-month-old group and the elder groups. The growth curves of the ear fibroblasts were ＂S-shaped＂ in different age groups. The cell proliferation of postnatal ear fi- broblasts, especially those from 2-, 4-, 6-, 12-month-old miniature pigs was significantly different from that of fetus ear fibroblasts （P〈0.05 or P〈0.01）. Two-month- and 4-month-old ear fibroblasts had a sig- nificantly higher proportion of G1 stage cells （85% to 91%） than those at 6 and 12 months （66% to 74%, P〈0.01）. The blastocyst rate of reconstructed embryos originating from newborn, 1-, 2-, 4-month-old donor pigs was 6.06% to 7.69% with no significant difference from that in fetus fibroblast group （8.06%）. It was concluded that 〈4-month-old adult Bama miniature pigs represent a better donor cell resource than elder pigs.
基金funded by the Project of Guangxi Science and Technology Basic Condition Platform Construction (11-31-09)
文摘[ Objective] The research aimed to discuss the relationship between the polymorphism of PPARy.2 gene and the susceptibility of type 2 diabetes mellitus （T2DM） in Guangxi Bama mini-pigs. [ Method] 24 Guangxi Bama mini-pigs were fed with high-fat and high-sucrose diet, and partial sequences of exon 2 of PPARy-2 gene were amplified by using PCR method. In addition, the contents of fasting blood glucose and insulin （INS） in Guangxi Bama mini-pigs were determined, and the glucose tolerance test （GTT） was also carried out. [ Result] There was one SNP site （19813A/G） Jn partial sequence of exon 2 of the cloned PPAFly-2 gene, and AA （7 pigs） and AG （17 pigs） genotype were detected. The contents of fasting insulin and 60-min blood glucose in GTT in AG-genotype Guangxi Bama mini-pigs were significantly higher than those of AA genotype （ P 〈0.05）, while the incidence of T2DM in AG-genotype Guangxi Bama mini-pigs （71.4%） was obviously higher than that of AA gen- otype （5.9%）. [ Conclusion] The polymorphism of 19813A/G in exon 2 of PPARy-2 gene was related with the susceptibility of T2DM in Guangxi Bama mini-pigs.
文摘Bama County。in the Guangxi Zhuang Autonomous Region in South China.is well known for a longer meanage of its population in China and the world over.It is called a 1and of longevity,and there are at present 66 centenarians in the county.
基金supported by grants from National Natural Science Foundation of China(No.81260423,No.81560523)
文摘Objective: To study the interaction between the polymorphisms of FOXO3A and the environmental factors in the longevity district of Bama county.Methods: Using Taqmaq probe,genotyping of FOXO3A gene was performed for 200 longevous elderly(aged 90-110)and 288 healthy controls(aged 48-89).The interaction between the polymorphisms of gene and environmental factors were analyzed by using conditional logistic regression.Results: Minor allele frequencies(MAF)of five known SNPs in FOXO3A gene were higher in cases than in controls(P<0.05).There was a positive interction between minority alleles of rs2764264 in FOXO3A gene and the environment factors in the longevity district(OR=11.08,γ=1.075).Conclusion: Five SNPs of FOXO3A gene would be closely correlated with human longevity in Bama.The interaction between the polymorphisms of FOXO3A genes rs2764264 and the environment factors in the longevity district had synergy to longevity.The longevity phenomenon in Bama probably was the result of the interaction between gene and environmental factors.
基金supported by the National Natural Science Foundation of China(31790410,32160781)。
文摘The gastrointestinal tract is essential for food digestion,nutrient absorption,waste elimination,and microbial defense.Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding of cellular diversity,functional heterogeneity,and their importance in intestinal tract development and disease.Although such profiling has been extensively conducted in humans and mice,the single-cell gene expression landscape of the pig cecum remains unexplored.Here,single-cell RNA sequencing was performed on 45572 cells obtained from seven cecal samples in pigs at four different developmental stages(days(D)30,42,150,and 730).Analysis revealed 12 major cell types and 38 subtypes,as well as their distinctive genes,transcription factors,and regulons,many of which were conserved in humans.An increase in the relative proportions of CD8^(+)T and Granzyme A(low expression)natural killer T cells(GZMA^(low)NKT)cells and a decrease in the relative proportions of epithelial stem cells,Tregs,RHEX^(+)T cells,and plasmacytoid dendritic cells(pDCs)were noted across the developmental stages.Moreover,the post-weaning period exhibited an up-regulation in mitochondrial genes,COX2 and ND2,as well as genes involved in immune activation in multiple cell types.Cell-cell crosstalk analysis indicated that IBP6^(+)fibroblasts were the main signal senders at D30,whereas IBP6^(−)fibroblasts assumed this role at the other stages.NKT cells established interactions with epithelial cells and IBP6^(+)fibroblasts in the D730 cecum through mediation of GZMA-F2RL1/F2RL2 pairs.This study provides valuable insights into cellular heterogeneity and function in the pig cecum at different development stages.
文摘Introduction: Pesticides are currently an essential component of agricultural production techniques for controlling pests and weeds. In Burkina Faso, non-compliance with good practice in the use of pesticides poses a real health problem for the population. This study examines the health risks associated with pesticide management in rice-growing areas. Material and Methods: A field survey was conducted in Bama, involving farmers, focusing on their socio-demographic characteristics, pesticide usage, and health effects. Cholinesterase levels were measured in subsample of farmers using a portable device. Data were analysed using Microsoft Excel, calculating means and percentages for various practices. Health consultations, protection methods, and pesticide management were studied. Erythrocyte acetylcholinesterase activity was compared before and after treatment. Data were categorised into classes based on inhibition levels, and correlation analyses determined relationships between variables such as age, years of experience, and cholinesterase activity. Results: The results indicate that rice cultivation is mainly carried out by a fairly young population, with nearly 63% being under the age of 50. Common poor practices in pesticide use include improper storage and reuse of leftover pesticides. Seven types of pesticides were identified, including organophosphates such as glyphosate, which was used in 26.7% of cases. This organophosphate has resulted in class B poisoning, causing a 30% - 50% reduction in erythrocyte cholinesterase activity. The health effects of pesticide use are felt by agricultural farmers through various symptoms of poisoning. Conclusion: To reduce the occurrence of pesticide poisoning, it is essential to launch information and awareness campaigns among the population and farmers to promote safe practices in pesticide use in Bama, Burkina Faso.
