基于BLI技术的BamAβ-barrel结合化合物检测方法的建立

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。

Approaches for Improvement of Competitiveness of Bama Rural Health-caring Tourism from the Deep Perspective of Tourism,China

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.

Cloning and Sequence Analysis of Adiponectin Receptor 1 and Receptor 2 cDNA from Guangxi Bama Mini-pig

[ 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.

石油烃厌氧降解基因masD和bamA实时荧光定量PCR方法的建立及应用

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。

Genomic and transcriptomic analyses reveal selection of genes for puberty in Bama Xiang pigs

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.

Heat Stress Upregulates the Expression of TLR4 and Its Alternative Splicing Variant in Bama Miniature Pigs

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.

Nucleus Transfer Efficiency of Ear Fibroblast Cells Isolated from Bama Miniature Pigs at Various Ages

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.

石油烃厌氧降解关键基因masD和bamA的实时荧光定量PCR检测方法与应用

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.

Correlation between the Polymorphism of PPARγ-2 gene and the Susceptibility of Type 2 Diabetes Mellitus in Guangxi Bama Mini-pigs

[ 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.

副溶血性弧菌外膜蛋白BamA重组表达及其免疫原性分析

副溶血性弧菌外膜蛋白BamA是β-桶组装BAM复合物的核心成分,参与细胞外膜蛋白运送和安装过程,是潜在的新型靶标抗原,目前其在免疫检测抗原和疫苗中的潜在价值尚未有研究报道。本研究通过生物信息学软件SnapGene和Protean分析BamA的序列并筛选出多肽,采用PCR扩增出外膜蛋白BamA的基因片段,构建重组质粒pET-28a(+)-BamA;经大肠杆菌BL21诱导表达BamA重组蛋白(90ku);多肽与BSA偶联,制备的抗原免疫BALB/c小鼠制备了血清。采用酶联免疫分析(ELISA)和蛋白质印迹法(Westernblot)测定BamA蛋白及多肽免疫血清对24株弧菌的交叉反应。ELISA测定结果表明,免疫血清对BamA蛋白的效价在121K以上,对副溶血性弧菌等弧菌的效价较弱(0.5K);免疫印迹结果显示,BamA蛋白血清与副溶血性弧菌CICC21617、CICC21618、杀岩龙虾弧菌和非O1型霍乱弧菌等弧菌属细胞裂解物中90ku左右的蛋白可发生特异性反应,而对费氏另类弧菌、嗜水气单胞菌和迟缓爱德华氏菌等非弧菌属无交叉反应。弧菌外膜蛋白BamA具有弧菌属保守性并可以诱导产生弧菌特异性抗体,多肽在菌体表面的暴露性可能受到其他抗原的影响,这给后续弧菌诊断抗原和疫苗抗原研究提供了依据。

Whole-genome sequencing study to identify candidate markers indicating susceptibility to type 2 diabetes in Bama miniature pigs

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.

Bama County: A Land of Longevity

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.

Study on the interaction between polymorphisms of FOXO3A gene and environmental factors in longevity at Bama county,Guangxi province

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.

Structural insights into Deinococcus radiodurans BamA:extracellular loop diversity and its evolutionary implications

Diderm bacteria,characterized by an additional lipid membrane layer known as the outer membrane,fold their outer membrane proteins(OMPs)via theβ-barrel assembly machinery(BAM)complex.Understanding how the BAM complex,particularly its key component BamA,assists in OMP folding remains crucial in bacterial cell biology.Recent research has focused primarily on the structural and functional characteristics of BamA within the Gracilicutes clade,such as in Escherichia coli(E.coli).However,another major evolutionary branch,Terrabacteria,has received comparatively less attention.An example of a Terrabacteria is Deinococcus radiodurans(D.radiodurans),a Gram-positive bacterium that possesses a distinctive outer membrane structure.In this study,we first demonstrated that theβ-barrel domains of BamA are not interchangeable between D.radiodurans and E.coli.The structure of D.radiodurans BamA was subsequently determined at 3.8Åresolution using cryo-electron microscopy,revealing obviously distinct arrangements of extracellular loop 4(ECL4)and ECL6 after structural comparison with their counterparts in gracilicutes.Despite the overall similarity in the topology of theβ-barrel domain,our results indicate that certain ECLs have evolved into distinct structures between the Terrabacteria and Gracilicutes clades.While BamA and its function are generally conserved across diderm bacterial species,our findings underscore the evolutionary diversity of this core OMP folder among bacteria,offering new insights into bacterial physiology and evolutionary biology.

细菌外膜蛋白BamA突变体构建及位点运动性分析

目的构建细菌外膜β桶状蛋白组装机器(BAM)核心组分BamA点突变原核表达载体,获得特定位点突变BamA蛋白,并分析该蛋白多肽易位相关(POTRA)结构域上G313位点运动特性。方法利用重叠延伸PCR(SOE-PCR)将BamA蛋白第690和700位天然半胱氨酸(C)突变为丝氨酸(S),将POTRA结构域上第313位氨基酸由甘氨酸(G)突为C,构建含BamA-C690S-C700S-G313C突变的原核表达载体pET22b-Strep-BamA。异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导突变蛋白表达后,利用Strep-Tactin Sepharose柱亲和层析方法对突变体蛋白进行纯化。甲烷硫代磺酸(MTSL)标记所得纯化蛋白的半胱氨酸后,进行电子顺磁共振(EPR)波谱检测,通过EPR波谱拟合获得旋转相关时间τ_(c),分析G313C氨基酸位点的运动特性。结果构建了BamA-C690S-C700S-G313C突变体原核表达载体,实现了突变体蛋白有效表达和纯化。G313C位点EPR波谱为单成分运动状态波谱,其τc值为(2.30±0.03)ns。结论该研究建立了BamA点突变原核表达和蛋白纯化方法,并获得BamA蛋白G313C运动性特征参数,为进一步研究BamA蛋白在外膜蛋白整合过程中的结构机制提供了数据。

Establishment of a model of Mycoplasma hyopneumoniae infection using Bama miniature pigs

Mycoplasma hyopneumoniae(M.hyopneumoniae),is the primary aetiological agent of enzootic pneumonia leading to chronic respiratory disease prevalent worldwide.Conventional pigs are the only animals used for pathogenicity studies and vaccine evaluations of M.hyopneumoniae.Considering that the challenge animals have better genetic stability and a smaller body size to operate with,an alternative experimental animal model of M.hyopneumoniae infection with Bama miniature pigs was established.Nine seven-week-old snatch-farrowed,porcine colostrum-deprived(SF-pCD)Bama miniature pigs and nine conventional pigs were randomly divided into two infected groups(Bama miniatureinfected(BI)and conventional-infected groups(CI),BI and CI,n=6)and two control groups(Bama miniature control(BC)and conventional control(CC)groups,BC and CC,n=3).Every piglet was tracheally inoculated with 5×10^(8) CCU/mL containing 10%suspension of a stock of frozen lung homogenate from SF-pCD pigs infected with virulent strain JS or sterilized KM2 medium.Typical lung lesions appeared in all infected pigs after necropsy,and the mean gross lung lesions was 17.3 and 13.7 in groups of BI and CI.Serum IgG and nasal sIgA antibody titres were increased significantly.Cilia shedding and mucus staining increased greatly in JS-infected bronchi.Obvious reddish gross lesions and M.hyopneumoniae antigen were detected,especially apparently observed in group of BI.Moreover,DNA copies of M.hyopneumoniae from bronchoalveolar lavage fluid(BALF)of each JS-infected piglet reached more than 10^(8),and M.hyopneumoniae could be re-isolated from each infected BALF.These results indicate that Bama miniature pigs could be used as an alternative and more maneuverable experimental infection model for M.hyopneumoniae and display typical clinical and pathological features consistent with those in conventional pigs.

母猪饲粮添加益生菌和合生元对子代巴马香猪肌肉氨基酸组成及生长发育相关基因表达的影响

[目的]本文旨在研究母猪饲粮添加益生菌和合生元对子代巴马香猪肌肉氨基酸组成及相关基因表达的影响。[方法]选用64头妊娠巴马香猪,随机分为对照组(基础饲粮)、抗生素组(50 g·t^(-1)维吉尼亚霉素)、益生菌组(200 mL·d^(-1)益生菌发酵液)和合生元组(500 g·t^(-1)低聚木糖+200 mL·d^(-1)益生菌发酵液),每组16头母猪,单栏饲养。母猪在整个妊娠期和哺乳期饲喂相应试验饲粮;仔猪断奶后,每窝选取2头仔猪,每组32头(8栏,每栏4头),饲喂基础饲粮。分别于65、95和125日龄每组选取8头采集股二头肌和腰大肌样品,测定其水解氨基酸组成和相关基因的表达量。[结果]与对照组相比,合生元组子代巴马香猪股二头肌粗蛋白、Gly、Ser、Ala、Asp、Glu、Pro和Ile含量,腰大肌Gly、Glu、Ser和His含量均显著增加(P<0.05);益生菌组子代巴马香猪股二头肌肌肉萎缩F-box蛋白32和生肌调节因子(MyoG)、腰大肌生肌因子5(Myf5)的表达水平显著上调(P<0.05),腰大肌肌球蛋白重链Ⅱb(MyHCⅡb)、股二头肌和腰大肌肌肉生长抑制素(MSTN)的表达水平显著下调(P<0.05);合生元组子代巴马香猪股二头肌Myf6、生肌分化因子和MyoG以及腰大肌Myf5和MyoG的表达水平显著上调(P<0.05),股二头肌MSTN的表达水平显著下调(P<0.05);益生菌组和合生元组子代巴马香猪股二头肌MyHCⅡx的表达水平显著下调(P<0.05);抗生素组子代巴马香猪腰大肌MyHCⅡx的表达水平显著上调(P<0.05),而股二头肌MyHCⅠ和腰大肌MyHCⅡa的表达水平显著下调(P<0.05)。[结论]母猪饲粮添加益生菌和合生元可改变子代巴马香猪肌肉氨基酸组成,调控生肌因子相关基因表达,上调氧化型肌纤维基因的表达、下调酵解型肌纤维基因的表达,从而有利于肉品质的改善。

巴马小型猪仰卧位的新型固定方法

目的 研究广西巴马小型猪仰卧位的固定方法以便于动物实验的开展。方法 选择行胸腔器官获取的广西巴马小型猪12头,接受全身麻醉后采用专用的固定装置进行仰卧位固定。结果 12头广西巴马小型猪在手术中体位得到很好的保持,未发生侧翻、移位等现象。结论 采用专用的固定装置对广西巴马小型猪进行仰卧位固定是一种简单有效的方法。

Single-cell profiling of the pig cecum at various developmental stages

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.

Cholinesterase Activity as an Indicator of Health Risks among Kou Valley Farmers

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.