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[Objectives] Porcine CD205, as a receptor specific to porcine dendritic cells (DC), played a key role in the antigen presentation. In this study, nanobodies against porcine CD205 CysR-FN Ⅱ protein were screened by phage display technology, which laid a foundation for the research of porcine CD205-targeted antigen presentation. [Methods] CysR-FN Ⅱ protein of porcine CD205 molecule prepared in the previous research was used for the immunization of alpacas. After subcutaneous immunization for 6 times, peripheral blood lymphocytes of alpaca were collected, their total RNA was extracted and cDNA was obtained by reverse transcription. The gene fragments encoding nanobodies were obtained by PCR amplification, and the porcine CD205 phage display nanobody library was constructed. Bio-spannings were performed by phage display technology and monoclonal colonies were randomly selected to screen specific nanobodies against porcine CD205 CysR-FN Ⅱ protein by Phage-ELISA. [Results] After 4 rounds of bio-spanning, a total of 12 porcine CD205 CysR-FN Ⅱ protein specific nanobodies were selected by Phage-ELISA and sequence analysis with their molecular weight about 17×10³. [Conclusions] This study successfully selected and obtained porcine CD205 CysR-FN Ⅱ protein specific nanobodies. [ABSTRACT FROM AUTHOR]

[Objectives] The aim of this study was to analyze the expression stability of reference genes in porcine granulosa cells (GC) during follicular development, and to further screen effective reference genes (RG)for the identification of key regulatory genes and their expression patterns. [Methods] Porcine preantral, antral, mature and atretic follicles were isolated and classified based on morphological observation with steroid hormone detection, and corresponding GC were collected. RT-qPCR was performed to detect the expression levels of nine candidate RG, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin beta (ACTB), tubulin alpha-1b (TUBA1B), ATP synthase f1 (ATP5F1), beta-2-microglobulin (B2M), ubiquitin C (UBC), hypoxanthine phosphoribosyltransferase 1 (HPRT1), ribosomal protein S3 (RPS3), and eukaryotic translation elongation factor 1 alpha (EEF1A1); ΔCt, geNorm, Norm Finder, Best Keeper and Ref Finder algorithms were used to analyze the expression stability of candidate RG in porcine GC during follicle development. [Results] The expression levels of candidate RG in porcine GC were significantly different, among which EEF1A1 and TUBA1B had higher expression levels, while HPRT1 and UBC had lower expression levels. ΔCt, NormFinder and Best Keeper analysis showed that RPS3 and ATP5F1 were the most stable RG, while GAPDH and HPRT1 were found to be more stable by geNorm analysis. Finally, the comprehensive weighted analysis by RefFinder showed that the stability of the candidate RG was ranked as follows:RPS3, ATP5F1, HPRT1, EEF1A1, GAPDH, ACTB, TUBA1B, B2M and UBC. Furthermore, the expression patterns of fertility-associated candidate genes (FSHR and NORFA) during follicular development were analyzed with different RG as controls, and results showed that the stability of RG was important for accurate identification of the expression and patterns of target genes. [Conclusions] RPS3 and ATP5F1 with higher expression stability could be used as effective RG for subsequent studies on sow follicular development, which also provide a theoretical basis for ideal RG selection to identify the key regulatory genes and their expression patterns during sow follicular development. [ABSTRACT FROM AUTHOR]

[Objective] The study aimed to select the optimal concentration of Mito-TEMPO. Meanwhile, the effect of Mito-TEMPO on the quality, antioxidation capacity, and mitochondrial function of frozen-thawed porcine spermatozoa were explored to provide a reference basis for the improvement of porcine spermatozoa cryopreservation. [Method] 0.5, 5.0, 50.0 and 500.0 µmol/L Mito-TEMPO were added to the frozen solution I in the experimental group. The concentration of the Mito-TEMPO suitable for the cryopreservation of porcine spermatozoa was screened by evaluating the kinetic parameters and viability of spermatozoa in each group. The plasma membrane intactness, acrosome intactness, DNA fragmentation index (DFI), reactive oxygen species (ROS) level, total antioxidant capacity (T-AOC), malondialdehyde (MDA) content, catalase (CAT) and total superoxide dismutase (T-SOD) activities, as well as ATP and mitochondrial respiratory chain complex IV content of freezing-thawing porcine spermatozoa were detected. [Result] Compared to the fresh group (Fgroup), the kinetic parameters and viability of spermatozoa were significantly reduced after freezing-thawing (P<0.05, the same as below). While the addition of Mito-TEMPO improved the kinetic parameters and viability of frozen-thawed spermatozoa to various degrees, and the effect of 50.0 µmol/L Mito TEMPO was the optimal. Compared to the F group, the plasma membrane and acrosome integrity, T-AOC, ATP and mitochondrial respiratory chain complex IV contents of frozen-thawed spermatozoa were significantly reduced, as well as CAT and T-SOD activities. DFI, ROS levels and MDA contents were significantly increased. Compared to the frozen control group (Cgroup), the plasma membrane integrity, асrosome integrity, T-AOC, as well as ATP and mitochondrial respiratory chain complex I, II and V contents of spermatozoa were significantly increased in the 50.0 µmol/L Mito-TEMPO group (M group). Moreover, the DFI, ROS levels and MDA contents were significantly reduced, and the CAT and T-SOD activities were significantly enhanced by the 50.0 µmol/L Mito-TEMPO. [Conclusion] Mito-TEMPO has a cryoprotective effect on porcine spermatozoa, and the optimal concentration is 50.0 µmol/L. In addition, 50.0 µmol/L Mito-TEMPO increases the quality of spermatozoa by improving the antioxidation capacity and mitochondrial function of porcine spermatozoa after freezing-thawing. [ABSTRACT FROM AUTHOR]

【Objective】Autoencoder, as one of the deep learning algorithms, offers unique advantages in reducing dimensionality and conducting interpolation analysis on single-cell transcriptome data. The study aims to assess the feasibility of applying the autoencoder AutoClass to early porcine embryo single-cell transcriptome data, and to investigate the impacts of different embryonic activation methods on key genes and signaling pathways.【Method】Single-cell transcriptome data were collected from early porcine embryos with 3 different activation modes (in vivo fertilization, in vitro fertilization, and orphaned females), and AutoClass was used to perform data quality control and interpolation analyses and evaluate AutoClass performance in conjunction with downstream analyses. In addition, an in-depth comparison of key genes and signaling pathways in the three types of embryos was performed by differential expression genes (DEGs) and functional enrichment analysis.【Result】After quality control and autoencoder data interpolation, the accuracy of clustering analysis was enhanced, leading to clear clustering among early embryos with different activation methods. QC alone only screened out 1 287 DEGs, while the number of DEGs increased to 11 523 after autocoder interpolation. Functional enrichment analysis further unearthed key biological processes and signaling pathways that were significantly different among the three different types of embryos, such as basic biological processes and cellular metabolism in porcine in vivo fertilized embryos, gonadal development and sex determination in porcine in vitro fertilized embryos and immune defenses and regulation of secretory pathways in orphaned embryos.【Conclusion】The autoencoder was adopted to improve the analytical accuracy of single-cell transcriptome data, whereby the key genes and signaling pathways affecting early embryonic development in three embryonic activation modes were revealed, which provided new ideas for further in-depth study of the molecular mechanisms underlying early embryonic development in porcine.

Copyright of Journal of South China Agricultural University is the property of Gai Kan Bian Wei Hui and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

【Objective】 Autoencoder, as one of the deep learning algorithms, offers unique advantages in reducing dimensionality and conducting interpolation analysis on single-cell transcriptome data. The study aims to assess the feasibility of applying the autoencoder AutoClass to early porcine embryo single-cell transcriptome data, and to investigate the impacts of different embryonic activation methods on key genes and signaling pathways. 【Method】 Single-cell transcriptome data were collected from early porcine embryos with 3 different activation modes (in vivo fertilization, in vitro fertilization, and orphaned females), and AutoClass was used to perform data quality control and interpolation analyses and evaluate AutoClass performance in conjunction with downstream analyses. In addition, an in-depth comparison of key genes and signaling pathways in the three types of embryos was performed by differential expression genes (DEGs) and functional enrichment analysis. 【Result】 After quality control and autoencoder data interpolation, the accuracy of clustering analysis was enhanced, leading to clear clustering among early embryos with different activation methods. QC alone only screened out 1 287 DEGs, while the number of DEGs increased to 11 523 after autocoder interpolation. Functional enrichment analysis further unearthed key biological processes and signaling pathways that were significantly different among the three different types of embryos, such as basic biological processes and cellular metabolism in porcine in vivo fertilized embryos, gonadal development and sex determination in porcine in vitro fertilized embryos and immune defenses and regulation of secretory pathways in orphaned embryos. 【Conclusion】 The autoencoder was adopted to improve the analytical accuracy of single-cell transcriptome data, whereby the key genes and signaling pathways affecting early embryonic development in three embryonic activation modes were revealed, which provided new ideas for further in-depth study of the molecular mechanisms underlying early embryonic development in porcine. [ABSTRACT FROM AUTHOR]

[Objectives]The aim of our study was to improve the efficiency of genome-editing in porcine embryonic fibroblasts (PEF) by Cas9/sgRNA system. [Methods]Two sgRNA were designed for the promoter region (first exon included) of porcine ROSA26 locus, and then they were constructed into PX459-cas9-puro plasmid to form PX459-cas9-sgRNA plasmids. The target sequence of sgRNA and their protospacer-adjacent motif (PAM) was constructed into RGS-CR to form RGS-sgRNA plasmid. The PX459-cas9-sgRNA and RGS-sgRNA plasmid system were combined to screen the sgRNA which had higher genome-editing efficiency, and this sgRNA was used to conduct subsequent transfection experiments on PEF. PEF cells were transfected with PX459-cas9-sgRNA2 plasmid and Cas9/sgRNA2,respectively. Genomic DNA was extracted from the transfected cells, and the targeting efficiency was measured by Sanger sequencing. [Results]PX459-cas9-sgRNA targeting ROSA26 sequence and RGS-sgRNA2 plasmids containing PAM were successfully constructed, and Sanger sequencing showed that the sequence of sgRNA1 and sgRNA2 was inserted correctly. RGS-sgRNA system showed that the editing efficiency of sgRNA2 was significantly higher than that of sgRNA1 (P＜0.01), and sgRNA2 showed no off-target in porcine genome, therefore, PX459-cas9-sgRNA2 and Cas9/sgRNA2 were used to compare the editing efficiency in PEF cells. The editing efficiency of PX459-cas9-sgRNA2 plasmid in PEF was 15.67%,and the editing efficiency of Cas9/sgRNA2 system was 33.33%. [Conclusions]Compared with PX459-cas9-sgRNA2 plasmid, Cas9/sgRNA2 can improve the targeting efficiency by about 1 fold, which improves genome-editing efficiency of pig somatic cell and is of great significance for promoting application of gene-editing in pigs. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

ObjectiveA sensitive and rapid method was developed for the determination of 17 β-adrenergic receptor blockers(β-blockers) by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS).MethodsThe β-blockers were extracted from porcine sample with acetonitrile, purified by solid phase extraction column Oasis Prime HLB. The analytes were then separated by the Endeavorsil C18 chromarographic column with methanol and 0.1% formic acid as gradient mobile phase. The ion fragment information analytes were obtained using ultra performance liquid chromatography-tandem mass spectrometry under multiple reaction monitoring mode and the isotope internal standards were used to quantify by calibration curves.ResultsThe linear ranges of 17 β-blockers were from 0.2 to 20 μg/kg, with correlation coefficient (R)>0.999. The limits of detection (LOD) were 0.05-0.07 μg/kg, and the limits of quantification (LOQ) were 0.15-0.2 μg/kg. The average recoveries of β-blockers(added into blank porcine samples at three concentration levels) ranged from 84.97% to 123.40%, and the relative standard deviation (RSD) was 0.41% to 13.94%.ConclusionThe method is simple, sensitive, accurate and is suitable for the confirmation and quantification of β-blockers in porcine samples.

[Objectives] This study aimed to analyze the molecular mechanism of porcine long non-coding RNA lincRNA-CXCL6 in the regulation of inflammatory response induced by lipopolysaccharide(LPS) and provide the new strategy for the defense against inflammatory damage caused by bacterial diseases in pigs. [Methods] The full-length sequence of lincRNA-CXCL6 was obtained by rapid amplification of complementary DNA(cDNA) ends, and the expression pattern of lincRNA-CXCL6 was detected by quantitative PCR(qPCR). Transcriptome sequencing was used to analyze the effect of lincRNA-CXCL6 on the cell signaling pathway after LPS stimulation based on the RAW264.7 cells stably expressing lincRNA-CXCL6. Western blot and qPCR were used to verify that lincRNA-CXCL6 regulated LPS-induced expression of inflammatory factors through the extracellular regulated protein kinases1/2(ERK1/2) pathway. [Results] The full length of lincRNA-CXCL6 was 985 bp, including two exons. The expression level of lincRNA-CXCL6 increased under the stimulation of LPS and Pam2CSK4, and its expression level gradually increased with the increase of LPS stimulation time, and it was only weakly expressed in the lung and small intestine in adult tissues of pigs. lincRNA-CXCL6 was mainly expressed in the cytoplasmic. The overexpression of lincRNA-CXCL6 inhibited the activation of interleukin 1β(IL-1β) and interleukin 6(IL-6) induced by LPS. Transcriptome data showed that compared with the control, lincRNA-CXCL6 overexpressing group showed 104 genes up-regulated and 165 genes down-regulated after LPS stimulated. Gene ontology(GO) analysis showed that 165 down-regulated genes were mainly involved in immune-related biological processes. lincRNA-CXCL6 negatively regulated the transcriptional expression of IL-1β and chemokines 17(CCL17) through the ERK1/2 pathway. [Conclusions] Porcine lincRNA-CXCL6 attenuated the expression of inflammatory factors mediated by LPS through the ERK1/2 signal pathway. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

BACKGROUND: Induced pluripotent stem cells are a cell source with multiple differentiation potentials. Since pig is a suitable model for human cardiovascular and metabolic diseases, induction of porcine pluripotent stem cells into endothelial cells will provide a support for creating a porcine cardiovascular disease model. OBJECTIVE: To establish a serum-free single layer culture method to induce porcine induced pluripotent stem cells into CD31 positive endothelial cells and to identify the obtained endothelial cells. METHODS: We used the serum-free single layer culture method to induce porcine pluripotent stem cells and human embryonic stem cells into endothelial cells under different combinations of CHIR99021, bone morphogenetic protein 4, basic fibroblast growth factor and vascular endothelial growth factor through three stages. The expressions of germ layer markers of the two kinds of stem cells during their differentiation were detected and compared. The endothelial cells obtained after the flow sorting were identified. RESULTS AND CONCLUSION: The endothelial cells differentiated from porcine induced pluripotent stem cells by a single layer culture method showed comparable morphological and functional properties to the immortalized porcine aortic endothelial cells, capable of taking up low-density lipoprotein and forming microvascular-like structure on matrigel. We found that the differentiation efficiency of porcine pluripotent stem cells was higher than that of human embryonic stem cells using the same method, which may be associated with the different expression patterns of lineage-specific markers in the two kinds of cells during the early differentiation. [ABSTRACT FROM AUTHOR]

Copyright of Journal of South China Agricultural University is the property of Gai Kan Bian Wei Hui and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Southern Agriculture is the property of Journal of Southern Agriculture and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

[Ｏｂｊｅｃｔｉｖｅｓ] Ｔｈｅ ｐａｐｅｒ ｗａｓ ａｉｍｅｄ ｔｏ ｃｒｅａｔｅ ａ ｍｅｔｈｏｄ ｆｏｒ ｔｈｅ ｉｓｏｌａｔｉｏｎ ｏｆ ｄｅｎｄｒｉｔｉｃ ｃｅｌｌｓ( ＤＣ) ｆｒｏｍ ｐｏｒｃｉｎｅ ｉｎｔｅｓｔｉｎａｌ ｍｕｃｏｓａｌ Ｐｅｙｅｒ’ｓ ｐａｔｃｈ(ＰＰ)ｉｎ ｖｉｔｒｏ. [Ｍｅｔｈｏｄｓ]Ｔｈｅ ｍｏｎｏｎｕｃｌｅａｒ ｃｅｌｌｓ ｗｅｒｅ ｏｂｔａｉｎｅｄ ｂｙ ｄｅｎｓｉｔｙ ｇｒａｄｉｅｎｔ ｃｅｎｔｒｉｆｕｇａｔｉｏｎ ａｎｄ ｔｈｅｎ ＤＣ ｗａｓ ｓｏｒｔｅｄ ｆｒｏｍ ｐｏｒｃｉｎｅ ｉｎｔｅｓｔｉｎａｌ ｍｕｃｏｓａｌ ＰＰ ｂｙ ｍａｇｎｅｔｉｃ ａｃｔｉｖａｔｅｄ ｃｅｌｌ ｓｏｒｔｉｎｇ( ＭＡＣＳ) ａｎｄ ｆｌｕｏｒｅｓｃｅｎｃｅ ａｃｔｉｖａｔｅｄ ｃｅｌｌ ｓｏｒｔｉｎｇ (ＦＡＣＳ)ꎬ ａｎｄ ｔｈｅｉｒ ｍｏｒｐｈｏｌｏｇｉｃａｌ ａｎｄ ｉｍｍｕｎｏｌｏｇｉｃａｌ ｃｈａｒａｃｔｅｒｉｓｔｉｃｓ ｗｅｒｅ ｉｎｖｅｓｔｉｇａｔｅｄ. [ Ｒｅｓｕｌｔｓ] Ｔｈｅ ｐｕｒｉｔｙ ｏｆ ＳＷＣ３ａ＋ ｃｅｌｌｓ ｅｎｒｉｃｈｅｄ ｂｙ ＭＡＣＳ ｗａｓ ９０.２％ꎬｗｈｉｌｅ ｔｈｅ ｐｕｒｉｔｙ ｏｆ ＳＷＣ３ａ＋ / ＭＨＣ￣Ⅱ＋ ｃｅｌｌｓ ｗａｓ ５０.５％. Ｔｈｅ ｐｕｒｉｔｙ ｏｆ ＳＷＣ３ａ＋ / ＭＨＣ￣Ⅱ＋ ＤＣ ｉｎ ｐｏｒｃｉｎｅ ｉｎｔｅｓｔｉｎａｌ ｍｕｃｏｓａｌ ＰＰ ｗａｓ ９５.６％ ｗｈｅｎ ｉｔ ｗａｓ ｓｏｒｔｅｄ ｂｙ ｆｌｏｗ ｃｙｔｏｍｅｔｒｙ. Ａｆｔｅｒ ２ ｄａｙｓ ｏｆ ｃｕｌｔｕｒｅꎬｔｈｅ ｃｅｌｌｓ ｈａｄ ａｇｇｒｅｇａｔｉｏｎ. Ｔｈｅ ｓｕｒｆａｃｅ ｏｆ ｔｈｅ ｃｅｌｌｓ ｗａｓ ｉｒｒｅｇｕｌａｒ ａｎｄ ｓｌｉｇｈｔｌｙ ｐｒｏｔｕｂｅｒａｎｔꎬａｎｄ ｈａｄ ｔｈｅ ａｂｉｌｉｔｙ ｔｏ ｓｔｉｍｕｌａｔｅ ｔｈｅ ｐｒｏｌｉｆｅｒａｔｉｏｎ ｏｆ ａｌｌｏｇｅｎｅｉｃ ｌｙｍｐｈｏｃｙｔｅｓ. [ Ｃｏｎｃｌｕｓｉｏｎｓ]Ｔｈｅ ｍｅｔｈｏｄ ｔｏ ｉｓｏｌａｔｅ ＤＣ ｆｒｏｍ ｐｏｒｃｉｎｅ ｉｎｔｅｓｔｉｎａｌ ｍｕｃｏｓａｌ ＰＰ ｉｓ ｅｓｔａｂｌｉｓｈｅｄꎬｗｈｉｃｈ ｍｉｇｈｔ ｃｏｎｔｒｉｂｕｔｅ ｔｏ ｆｕｒｔｈｅｒ ｓｔｕｄｙ ｏｎ ｔｈｅ ｒｏｌｅ ｏｆ ＤＣ ｉｎ ｐｏｒｃｉｎｅ ｉｎｔｅｓｔｉｎａｌ ｍｕｃｏｓａｌ ＰＰ. [ABSTRACT FROM AUTHOR]

Objective To explore the feasibility of the adjustable brace asymmetric tethering in concave side for establishing a porcine scoliosis model．Methods Six minority piglets （aged 8-10-week-old，weight 8-10 kg） were selected and the adjustable brace asymmetric tethering in concave side were applied during the procedure．Roentgenography was performed before and immediately after the operation，and 1，2 and 3 months after the procedure．Cobb angles were measured based in the plain radiograph．Results A piglet died for narcotic drug overdose and the other 5 pigs succeeded in modeling．The cobb angles were（9.0±1.6）°in one month， （11.8±1.3）°in two months and（21.6±2.4）°in three months after the operation . Conclusion It's is an effective way to establish the porcine model of rapidly progressive structural scoliosis by the adjustable brace asymmetric tethering in concave side．It avoids the damage to the spinal elements and could be an ideal model for further study on corrective techniques. [ABSTRACT FROM AUTHOR]

Objective: To investigate the repair method of type Ⅱc injury in the lateral meniscus popliteal tendon area based on the porcine knee joint., Methods: Eighteen commercially available fresh porcine knee joints were randomly divided into 3 groups ( n =6). After preparing a type Ⅱc injury in the lateral meniscus popliteal tendon area, and the anterior (group A), posterior (group B), or anterior and posterior (group C) of the popliteal hiatus (PH) was sutured by vertical mattress. The tension meter was used to apply gradient tensions of 2, 4, 6, 8, and 10 N along the tibial plateau horizontally, respectively, to pull the midpoint of the lateral meniscus popliteal tendon area. The displacement values before modeling, after modeling, and after suture were recorded. The reduction value of lateral meniscus displacement and reduction rate after suture were calculated and compared between groups., Results: There was no significant difference between groups ( P >0.05) in the displacement values before modeling, after modeling, and after suture under different tensions. There was no significant difference between groups A and C ( P >0.05) in the reduction value of lateral meniscus displacement and reduction rate after suture under different tensions. The reduction value of lateral meniscus displacement and reduction rate after suture in group B were lower than those in groups A and C. The reduction value of lateral meniscus displacement under tension of 2 N and the reduction rates under tensions of 2, 4, and 6 N between groups A and B showed significant differences ( P <0.05). The reduction value of lateral meniscus displacement and the reduction rate under tensions of 2, 4, and 6 N between groups B and C showed significant differences ( P <0.05)., Conclusion: Suturing the anterior area of PH is the key to repairing type Ⅱc injury of lateral meniscus popliteal tendon area.

[Objectives] The purpose of this study was to investigate the effects of Aurora-A inhibition on maturation,spindle configuration of porcine oocytes and potential roles.[Methods] Porcine GV oocytes were randomly assigned into groups to culture in vitro. Each group was added with 0, 1, 3 and 5 μmol·L-1 MLN8054,an Aurora-A inhibitor, respectively. Maturation of oocytes and spindle structural alteration were observed. Phosphorylated Aurora-A and TACC3 (transforming acidic coiled-coil proteins) expression were analyzed by Western blot.[Results] The maturation rate of porcine oocytes was significantly inhibited when treated with the Aurora-A inhibitor (P＜0.05),mainly arresting at Pro-MⅠand MⅠ-stage. Monopolar or multipolar spindles were observed at MI-stage. The percentages of abnormal spindle morphology for MLN8054-treated groups were significantly increased as compared to the control (P＜0.05). The pAurora-A and pTACC3 expressions of oocytes after Aurora-A inhibition were significantly decreased (P＜0.05).[Conclusions] Maturation of oocytes and spindle configuration were affected and pAurora-A and its downstream protein pTACC3 expressions were inhibited by MLN8054 treatment. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Veterinary Science / Zhongguo Shouyi Xuebao is the property of Chinese Society of Veterinary Science and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Veterinary Science / Zhongguo Shouyi Xuebao is the property of Chinese Society of Veterinary Science and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Chinese Journal of Veterinary Science / Zhongguo Shouyi Xuebao is the property of Chinese Society of Veterinary Science and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163 . Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.

The purpose of this article was to investigate the distribution of dendritic cells (DC) in healthy porcine tonsil. In this study, three different anti-porcine DC antibodies and fluorescence microscopy as well as confocal microscopy were used to examine the distribution of porcine tonsil DC on 10 crossed-bred(Duroc×Landrace×Yorkshine) pigs aged 2 months. The results showed that: DC in porcine tubal tonsil and soft palatine tonsil could co-express CD11b with CD16 or CD11b with MHCII; The number of double-stained DC in porcine tonsil lamina propria were three times higher than lymphoid follicles; There were only small amounts of DC between the epitheliums, and DC in soft palatine tonsil were hardly be found; The majority of CD11b+DC were observed under the epithelium of tonsil; A rare population of entending cytoplasmic processes between epithelium had been identified as a possible correlate for antigen uptake. There were cetanin number of DC in lamina propria of porcine tubal tonsil and soft palatine tonsil for antigen absorption and immune activation which provided a theoretical basis for intranasal and oral immunization. [ABSTRACT FROM AUTHOR]

The article focuses on the study which investigates the efficacy of valproic acid (VPA) on the developmental competence of porcine handmade cloning (HMC) reconstructed embryos from different donor cells. The study used histone deacetylase inhibitors VPA. The results of the study showed that the blastocyst formation rate was 48. 92% and the total blastosphere cellular rate was 80. 33.

The article presents a study which aims to clone and analyze the sequence of promoters in pig liver carboxylesterase 1 (PLE1).

The article presents a study on the intracytoplasmic injection of porcine, buffalo and cynmolgus monkey spermatozoon into porcine oocytes in China. Result shows that in the first experiment, the rate of pronuclear formation of injecting porcine spermatozoa group was higher compared to other groups while in the second experiment there was significant difference among the three groups. The study concluded that the pronuclear and blastocyt formation were observed among the three groups.

The article presents a study regarding the gene expression of a mature porcine interferon-alpha (PoIFN-α) fused with C-terminal 6 x Histidine tag using the baculovirus pFastBac™ I vector. The recombinant proteins of the gene expression were detected by Western blotting analysis and immunofluorescence assays. The results of the study indicate the therapeutic effect of PoIFN-α which inhibits the replication of porcine reproductive and respiratory syndrome virus (PRRSV).

The article focuses on a study related to examining the factors responsible for procine oocytes in vitro fertilization (IVF) efficiency. Topics discussed include the effect of cumulus-oocytes complexes on in vitro fertilization cleavage rate and blastocyst rate, the determination of the influence of sperm floating time on IVF and the effect of gametes co-incubation time on IVF.

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With the advancements of stem cells and regenerative medicine, interspecies chimera has become a hot topic and will pave a new way of providing donor sources in organ transplantation. However, the interspecies chimera is confronted with a number of scientific questions and technical obstacles, including selections of appropriate embryonic stage and appropriate culture medium; those factors will deeply influence the developmental balance between donor cells and receptor embryos. Due to its relatively rapid reproductive cycle and similar organ size to human's, porcine is a very potential donor candidate to study these questions. To compare the development and chimeric efficiency of interspecies embryos, we tested and evaluated three different culture systems, PZM-3 (Porcine zygotic medium), culture medium for iPSCs (N2B27) and 3.5 h of N2B27 before PZM-3 (N2B27(3.5 h)), and two different embryonic stages, 8-cell and blastocyst in mouse-porcine chimeric embryos using parthenogenetically activated porcine embryos and mouse induced pluripotent stem cells (miPS). The results showed that, PZM-3 was beneficial for both development of chimeric embryos and miPSCs proliferation in porcine embryos in the 8-cell injection group. After early blastocyst injection, the chimeric efficiency did not appear significantly different among the three culture systems but was lower than 8-cell injection. In summary, the results suggest that 8-cell injection and PZM-3 culture medium are more beneficial to the in vitro development and chimeric efficiency of mouse-porcine chimeric embryos.