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。

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.

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.

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。

含血必净注射液的供心保存灌注液对ECMO支持下离体空跳猪心心肌保护作用观察

目的观察含血必净注射液的供心保存灌注液对体外膜肺氧合(ECMO)支持下离体空跳猪心的心肌保护作用,并探讨其可能的机制。方法选取健康成年广西巴马小型猪12只,随机分成血必净组和对照组,每组6只。血必净组和对照组均采用ECMO支持猪离体空跳心脏,分别用含50 mL血必净注射液、生理盐水的去白细胞氧合血连续灌注,维持心脏空跳8 h。灌注0 h(T1)、2 h(T2)、4 h(T3)、6 h(T4)、8 h(T5),记录两组心率、灌注压、灌注流量,采用ELISA法检测各时间点灌注液炎症相关指标白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平。使用全自动生化分析仪检测两组全身肝素化后且灌注前(T0)及T1~T5时间点灌注液心肌损伤标志物cTnT(cTnT)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)水平。透射电镜下观察两组T5时间点的心肌组织超微结构。结果两组T1~T5时间点心率、灌注压、灌注流量比较差异均无统计学意义(P均>0.05)。与同组T1时间点比较,对照组T3~T5时间点灌注液IL-1β、IL-6、TNF-α、cTnT、CK、CK-MB、LDH水平均升高,血必净组T4~T5时间点灌注液cTnT、CK、CK-MB、LDH水平均升高(P均<0.05);血必净组T3~T5时间点灌注液IL-1β、IL-6、TNF-α、cTnT水平均低于对照组,T2~T5时间点灌注液CK、CK-MB、LDH水平均低于对照组(P均<0.05)。心肌组织超微结构:对照组心肌组织中重度退行性变、脂肪变,细胞内基质大面积溶解,肌纤维结构排列紊乱、大量断裂,线粒体大多肿胀严重、大小不均,细胞膜溶解、破损;与对照组比较,血必净组细胞结构损伤相对较轻,心肌细胞轻度肥大,线粒体轻度肿胀、大小均一,肌纤维粗细均匀、排列整齐。结论含血必净注射液的供心保存灌注液对ECMO支持下离体空跳猪心具有心肌保护作用,其机制可能与抑制炎症反应、减轻心肌细胞线粒体损伤有关。

基于转录组测序的巴马香猪和杜洛克猪骨骼肌差异lncRNA分析

【目的】通过对巴马香猪和杜洛克猪背最长肌组织进行转录组测序鉴定差异表达lncRNA,筛选与骨骼肌发育形成相关的候选基因,为明确lncRNA对骨骼肌生长发育的调控机制提供理论基础。【方法】采集12月龄的巴马香猪和杜洛克猪背最长肌组织进行转录组测序,以错误发现率(FDR)<0.05且|log2Fold Change|>1为标准筛选差异表达基因(DEGs)和差异表达lncRNA,并进行实时荧光定量PCR验证,预测差异表达lncRNA靶基因并进行GO功能注释和KEGG信号通路富集分析,选取表达差异最大的lncRNA构建其与靶基因互作网络。【结果】在巴马香猪和杜洛克猪背肌间共鉴定出6316个DEGs和675个差异表达lncRNA;GO功能注释分析结果表明,差异表达lncRNA靶基因主要涉及代谢过程、肌肉组织发育及骨骼肌细胞增殖和分化等生物过程;KEGG信号通路富集分析结果表明,差异表达lncRNA靶基因在Hippo和Wnt信号通路显著富集(P<0.05),说明差异表达lncRNA与骨骼肌细胞的增殖、自噬和分化相关;对表达差异最大的lncRNA-MSTRG.16703进行实时荧光定量PCR验证,发现其在巴马香猪中的相对表达量极显著高于杜洛克猪(P<0.01),与转录组测序结果一致。lncRNA-MSTRG.16703与靶基因互作网络分析结果表明,上调靶基因包括QKI、MBNL1和YBX2,QKI和MBNL1在均与可变剪接相关。【结论】在巴马香猪和杜洛克猪间发现的差异表达lncRNA是调控骨骼肌发育的候选基因,其靶基因主要富集在Hippo和Wnt等骨骼肌发育相关信号通路。lncRNA-MSTRG.16703的表达差异最大且在巴马香猪中上调,其靶基因在骨骼肌细胞增殖分化和肌纤维形成中有重要调控作用。

“酶、植物提取物和益生菌”复方制剂对巴马香猪生长性能、血清生化指标和抗氧化指标的影响

探讨“酶、植物提取物和益生菌”复方制剂对巴马香猪生长性能、血清生化指标和抗氧化指标的影响,为“酶、植物提取物和益生菌”复方制剂的科学应用提供参考。试验选取健康状况相近的3月龄巴马香猪60头,平均体重15 kg左右,公母随机,分为对照组、试验1组和试验2组,每组20头巴马香猪,每组4个重复,每个重复5头。其中对照组饲喂基础日粮,试验1组饲喂基础日粮+100 g/kg植物提取物,试验2组饲喂基础日粮+100 g/kg“酶、植物提取物和益生菌”复方制剂,试验为期28 d。测定各组猪只的生长性能、血清生化指标和抗氧化指标。结果表明,“酶、植物提取物和益生菌”复方制剂能提高巴马香猪的生长性能、机体免疫力和抗氧化能力。

舒泰复合右美托咪定两种给药方案对巴马小型猪麻醉效果观察

【目的】探究不同舒泰复合右美托咪定给药方案对巴马小型猪麻醉效果的影响。【方法】将8头巴马小型猪随机均分为静脉注射(Ⅳ)组和肌内注射(IM)组,麻醉间隔期为14 d。Ⅳ组耳缘静脉注射舒泰3.3 mg/kg+右美托咪定8μg/kg;IM组耳后颈部肌内注射舒泰4.6 mg/kg+右美托咪定11.2μg/kg。在麻醉过程中对巴马小型猪的麻醉时期、麻醉效果、生理指标及生物反射进行监测。【结果】Ⅳ组麻醉诱导时间、维持时间、苏醒时间分别为3.75、85.75和30.25 min。IM组麻醉诱导时间、维持时间、苏醒时间分别为7.13、107.00和41.00 min。Ⅳ和IM组分别可维持50和65 min外科麻醉期。IM组80~110 min时麻醉效果评分显著高于Ⅳ组(P<0.05)。麻醉过程中两种方案的体温(T)下降,心率(HR)和血氧饱和度(SpO_(2))先降低后升高,呼吸频率(RR)、收缩压(SBP)、舒展压(DBP)和平均动脉压(MAP)先升高后降低。IM组50~80 min时RR显著高于Ⅳ组(P<0.05)。IM组10~15 min时SBP显著低于Ⅳ组(P<0.05)。Ⅳ和IM组生物反射完全抑制时间分别为45和50 min。【结论】舒泰复合右美托咪定对巴马小型猪麻醉效果良好,诱导迅速,维持时间长,苏醒平稳,对巴马小型猪生理指标和生物反射影响较小,其中IM组麻醉方案比Ⅳ组更适合临床外科手术和科研教学使用。

乡村振兴背景下事业单位类型新型研发机构发展现状案例分析——以巴马瑶族自治县乡村振兴研究院为例

为了探究事业单位类型新型研发机构在乡村振兴背景下利用科技创新助力乡村振兴的积极作用,文章以中国农业科学院深圳农业基因组研究所和巴马瑶族自治县人民政府共同创建的新型研发机构——巴马瑶族自治县乡村振兴研究院为例,详细介绍了该机构的建设背景、科技支持案例,分析了该机构目前的发展状况和面临的挑战,并从增强科技成果转化意识、健全科技人才激励机制、促进自身供血体系建设等方面提出了建议,旨在为事业单位类型新型研发机构的发展提供参考和借鉴,推动其更好地服务于乡村振兴事业。

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.

丘陵地区重要集镇预警雨量值计算技术研究——以广西巴马县燕洞镇为例

预警雨量值计算是山洪灾害防治非工程措施中的重要环节。为研究丘陵地区重要集镇预警雨量值计算方法,选取广西巴马县燕洞镇作为典型区域,通过设计暴雨、设计洪水、水位流量关系转换等方法,确定了预警时段、预警阈值等关键结论。

乡村振兴视域下基于AHP与模糊综合评价法的特色小镇建设绩效评价研究

全面推进乡村振兴战略背景下,特色小镇成为促进经济增长的重要驱动力。当前,特色小镇建设面临集聚功能较差、区域治理效能难凸显、生态效益潜力待发掘和组织协调能力需提升等复合生态发展结构化困境。该文在梳理和借鉴已有研究成果上,以乡村振兴为视角,构建乡村振兴、经济状况、产业发展、旅游资源、交通服务和政策支持6项一级指标,农业发展、农村建设、农民增收等24项二级指标的特色小镇建设绩效评价模型,并以河池市巴马县甲篆康养特色小镇建设绩效开展实证研究,采用AHP计算各级指标权重,运用模糊综合评价法作出总体评价。模糊综合评价结果表明,该特色小镇建设绩效评价总体“良”,由结果分析还提出落实乡村政策、完善交通服务、加强组织协调等建议。

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.

智能针灸机器人针刺可行性及安全性评价实验动物平台建立的探索

目的本研究旨在探索智能针灸机器人针刺可行性及安全性评价的实验动物平台,为后续进一步开展研究奠定基础。方法将6只2月龄广西巴马小型猪作为实验对象,进行麻醉后针刺验证。首先进行人工针刺验证,每只实验动物选取6个穴位,行提插捻转手法后留针20 min,并进行自身前后对照。实验共进行28 d,每2 d进行1次实验,共10次。人工针刺验证后在已选6个穴位的基础上每个穴旁10 mm处取一点,共12点,使用针灸机器人智能针刺模块对实验动物进行不同频率及角度的针刺操作进一步验证动物平台的稳定性及可行性。结果血常规中安全性相关指标均无异常,较自身前后对比稳定。血生化指标无异常,较自身前后对比稳定。在实验全程监测中动物的平均心率为每分钟124次。动物的平均血压为87/36 mmHg。巴马小型猪在室温25℃下的平均体温为36℃。体温在实验过程中及实验前后均无明显的变化。在搭建的实验平台基础上,针灸机器人智能针刺模块针刺手法操作预实验均顺利完成,实验过程中未出现针刺相关异常情况如弯针、断针、滞针等,实验动物未出现明显异常。结论本研究借鉴了已有的小型猪实验方法,初步建立起了稳定的智能针灸机器人针刺可行性及安全性评价实验动物平台,为进一步智能针灸机器人的相关研究开展奠定基础。

火麻的经济与文化:基于巴马火麻产业的调研

本文基于巴马火麻产业的调研,从火麻种植、加工、销售产业链的发展现状,分析火麻产业发展存在的困难,并针对这些困难,提出促进火麻“长寿价值”与民族文化融合发展,养生需求推动火麻产业发展等繁荣火麻产业发展的路径。

巴马火麻油对大鼠抗氧化功能的影响

为研究巴马火麻油对大鼠抗氧化功能的影响,依据《关于印发抗氧化功能评价方法等9个保健功能评价方法的通知》(国食药监保化[2012]107号)附件1抗氧化功能评价方法,采用巴马火麻油对成年大鼠进行实验,设溶媒对照组、空白对照组及火麻油高、中、低3个剂量组,连续灌胃30 d。结果表明,受试样品组肝组织匀浆和血清中的丙二醛低于溶媒对照组,差异显著(P<0.05),肝组织匀浆中的超氧化物歧化酶活性极显著高于溶媒对照组(P<0.01);受试样品组肝组织匀浆和血清中的还原型谷胱甘肽含量高于溶媒对照组,差异极显著(P<0.01)。这表明在本实验条件下,动物实验的脂质过氧化产物、抗氧化酶、抗氧化物质3项指标阳性,表明火麻油具有抗氧化功能。

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.