The environment of the Qinghai-Tibet Plateau is mainly manifested in low oxygen and low pressure, cold and dry, strong ultraviolet rays and food shortage. The plateau domestic animals can live in this environment after long-term selection and cultivation, and their organisms have formed unique adaptive characteristics to the plateau. Due to complicated characteristics of plateau domestic animals adaptive evolutionary system, the comprehensive and systematic analysis of adaptive molecular mechanism has not been completed. With the development of molecular biology and bioinformatics, the genome assembly and functional annotation of the plateau domestic animals have been accomplished, and then the work of genome, transcriptome and proteome has been gradually carried out. Since then, a series of key candidate genes for plateau animals environmental adaptability have been identified. These provided further support for the analysis of the molecular mechanisms in the environment adaptation of plateau domestic animals. This review takes Tibetan chicken (Gallus gallus), Tibetan pig(Sus scrofa), yak(Bos grunniens), Tibetan goat(Capra hircus), Tibetan sheep(Ovis aries), Tibetan horse(Equus caballus) and Tibetan mastiff(Canis lupus familiaris) as the classification unit, their anatomical structure of tissues and organs, blood physiological and biochemical indexes and molecular genetic mechanism are discussed, respectively, and the future research of plateau adaptive evolution are envisaged, which will lay a theoretical foundation for the new variety breeding in the plateau high-cold and hypoxic regions.

Glycopeptide antibiotics, especially vancomycin, are effective drugs for clinical treatment of multiple drug-resistant Gram-positive bacteria such as methicillin-resistant Staphylococcus aureus (MRSA). With the development of bacterial drug resistance, the problem of glycopeptide antibiotic resistance is becoming more and more serious, so it is very urgent to find and develop new compounds for clinical treatment. This paper reviews the action and resistance mechanism of glycopeptide antibiotics, newly approved glycopeptide antibiotics, chemical modification and new screening and development methods, in order to provide ideas for the development of glycopeptide antibiotics.

Amino acids are one of the most important dietary components in piglets, but the traditional classification of amino acids is defective. Functional amino acids are a redefinition of traditional amino acids and play an important role in animal immune function.This paper summerizes the metabolic effects of functional amino acids such as arginine, glutamate, glutamine, branched-chain amino acids, tryptophan, glycine, cysteine and proline in the intestinal tract of weaned piglets and their effects on intestinal health. The mechanism of functional amino acids to repair intestinal damage is also elucidated. In order to provide a basis for functional amino acids application in intestinal injury repair of weaned piglets.

In recent years, new progress has been made in the research of glycoprotein gE of alphaherpesvirus in virus intercellular transmission, nervous system infection, and immune evasion. gE is involved in syncytium formation, anterograde and retrograde neurotransmission, and is also the first reported viral protein that inhibits the activation of plasmacytoid dendritic cells to produce type I interferon. This summary reviews the relationship between gE and virulence to provide useful information for the further study of gE.

The purpose of this study was to screen candidate genes affecting pig body height traits by using genome-wide copy number variation regions (CNVRs) association analysis and genome-wide quantitative trait loci (QTLs) mapping. CNVcaller software was used for rapid detection of genome copy number variations based on resequencing data of the Large White×Min pig F2 population constructed. Mixed-linear model (MLM) with gender and parity as fixed effects was used to perform copy number variation genome-wide association study (CNVR-GWAS) of body height traits. The software R/qtl was used for QTL analysis and the permutation test (PT) was used for inspection. The results of CNVR-GWAS and QTL were jointly annotated, combined with GO enrichment and KEGG pathway analysis, the loci and genes affecting pig body height were excavated. Candidate genes were verified by real-time quantitative polymerase chain reaction (qPCR) method. The results showed that there were 3 099 CNVRs in the whole genome of the population. Among them, two CNVRs were significantly associated with body height trait in the whole genome level, and they located at 25 358 001-26 696 400 bp (CNVR1) and 54 087 201-54 090 000 bp(CNVR2) on chromosome 7, respectively. The results of mixed linear model analysis showed that the increase of copy number of CNVR1 (P<0.01) and the loss of copy number of CNVR2 (P<0.01) had significant effects on pig body height. Two significant QTLs (BH-1 and BH-2, respectively)affecting pig body height were also found at genomic level, of which BH-2 had a greater impact on body height trait. There were one olfactory receptor gene OR12D3 and 18 unannotated genes in the overlap region of CNVR1 and BH-2. And the verification results of qPCR indicated that the copy number variation of OR12D3 was consistent with the results inferred by mixed linear model statistics. The copy number variation of the OR12D3 gene was inferred to be associated with pig body height trait.

This experiment aimed to obtain the differentially expressed genes before and after the adipogenic differentiation of Jianzhou Da'er goat intramuscular preadipocytes by the RNA-seq technology, and acquire the related signaling pathways and key candidate functional genes by bioinformatics analysis. The 7-day-old healthy male Jianzhou Da'er goat was used (n=3) and their intramuscular preadipocytes were isolated by collagenase digestion. Illumina platform was used to carry out the high-throughput sequencing to two groups of cDNA samples (n=3) of Jianzhou Da'er goat intramuscular preadipocytes and intramuscular adipocytes induced for 5 day, respectively. The differentially expressed genes were obtained according to the thresholds of|log₂fold change|>0 and P<0.05. The differentially expressed genes were enriched to GO terms and KEGG pathways using the clusterProfiler R. Ultimately, the qRT-PCR technology was performed to detect the change of expression levels of candidate functional genes before and after differentiation of preadipocytes. The results showed that there were 7 916 differentially expressed genes in the RNA-seq results during the adipogenic differentiation of Jianzhou Da'er goat intramuscular preadipocytes, including 4 143 up-regulated genes that were enriched to 305 KEGG pathways, including oxidative phosphorylation, ribosome synthesis, and citrate cycle pathways, etc., and 3 773 down-regulated genes that were enriched to 303 KEGG pathways, including thyroxine signaling pathways, and so on; In the results of GO function enrichment analysis, the biological process was 52.8% of GO terms, the cellular component was 13.4%, and the molecular function was 33.8%. The qRT-PCR results showed that the trends of UCP3, ACACB, ACOT11, ACOX3, APOA1, and WISP2 expression during the adipogenic differentiation of intramuscular preadipocytes were consistent with the results of RNA-seq, which indicated that these genes were the suitable candidate functional genes for further research. This study obtained the differentially expressed genes during the adipogenic differentiation of Jianzhou Da'er goat intramuscular adipocytes and verified 6 candidate functional genes. The results of this study provided the basic data and systematic information for elucidating the molecular regulatory networks during the adipogenic differentiation of meat goat intramuscular adipocytes.

In order to understand the genetic trends of Chinese Merino sheep (Xinjiang type) in recent years and to explore the genetic relationship between wool and reproductive traits, it is necessary to further study the heritability of these traits and their relationship. The records of wool production performance of 9 428 individuals (from 1985 to 2018) and the records of reproduction performance of 5 887 individuals (from 1987 to 2018) for Chinese Merino sheep (Xinjiang type) were collected from Juxin fine wool sheep breeding professional cooperative of Emin county. Using a single trait model by BLUPF90 software combined with Gibbs sampling method estimated the variance components and heritabilities for wool (fineness, quality, total fleece grade, staple length, greasy fleece weight and body weight pre-shearing)and reproduction traits (number of inseminations, gestation length, litter size, total number of lambs born), and using two-trait model estimated the genetic correlation and phenotypic correlation between wool and reproductive traits. The results showed that estimates of heritabilities for fineness, quality, total fleece grade, staple length, greasy fleece weight and body weight pre-shearing were 0.471±0.020, 0.088±0.030, 0.114±0.018, 0.426±0.025, 0.328±0.041, and 0.317±0.046, respectively; estimates of heritabilies for number of inseminations, gestation length, litter size and total number of lambs born were 0.056±0.009, 0.022±0.010, 0.120±0.018, and 0.163±0.016, respectively; estimates of genetic correlation between wool traits and litter size, total number of lambs born varied from -0.031 to 0.286, body weight pre-shearing showed the strongest relation to litter size (0.286) followed by total number of lambs born (0.204), fineness were negatively related to litter size (-0.143) and total number of lambs born (-0.048); estimates of phenotypic correlation between wool traits and litter size, total number of lambs born varied from -0.210 to 0.216, staple length showed the strongest relation to total number of lambs born (0.216), fineness were negatively related to litter size (-0.137) and total number of lambs born (-0.210). The results of this study found that there was a certain relationship between wool and reproduction traits. It can provide a data basis for the future development of Chinese Merino sheep breeding program, and provide a theoretical basis for the selection of fine wool sheep with good quality, high yield and good reproductive performance, so as to further improve the economic benefits of fine wool sheep industry.

The aim of this study was to explore the function of the retinoic acid receptor responder 1 gene in Qianbei Ma goat. In this study, the complete sequence encoded by RARRES1 gene of Qianbei Ma goat was obtained by PCR amplification. The bioinformatics method was used to analyze the physical and chemical properties, higher protein structure, hydrophobicity, amino acid homology, subcellular localization of RARRES1 protein, and the phylogenetic tree was constructed. At the same time, qRT-PCR was used to detect the expression level of RARRES1 gene in different tissues of monotocous and polytocous of Qianbei Ma goat, and then the eukaryotic expression vector pEGFP-N3-RARRES1 and pGPH1/GFP/Neo-RARRES1 RNA interference vector were constructed and transfected into the follicular granulosa cells of Qianbei Ma goat. The effect of the eukaryotic expression vector pEGFP-N3-RARRES1 and pGPH1/GFP/Neo-RARRES1 RNA interference vector on the expression of RARRES1 and candidate genes for fecundity BMPR-IB mRNA were detected by qRT-PCR at cell level. qRT-PCR results showed that the expression of RARRES1 mRNA was the highest in the ovary, the expression level in ovarian tissue of Qianbei Ma goat in the monotocous group was extremely significantly higher than that in the polytocous group(P<0.01). Bioinformatics analysis showed that the RARRES1 gene encoded 289 amino acids, RARRES1 was a hydrophilic protein localized in the cytoplasm. The secondary structure analysis of the protein showed that it was mainly composed of α-helix and random coil. Its tertiary structure predicted was consistent with the secondary structure. The results of homology and phylogenetic tree showed that the RARRES1 gene sequence of Qianbei Ma goat had high homology with sheep and cattle, and their genetic distance was closer, the homology with chicken was the lowest and the genetic distance was the farthest. After double enzyme digestion and sequencing verification, the eukaryotic expression vector pEGFP-N3-RARRES1 and pGPH1/GFP/Neo-RARRES1 interference vector were successfully constructed. After the vectors were transfected into follicular granulosa cells of Qianbei Ma goat, compared with the blank control, recombinant plasmid pEGFP-N3-RARRES1 extremely significantly increased the expression of RARRES1 and BMPR-IB (P<0.01). Among the interfering vectors, the recombinant plasmid pGPH1/GFP/Neo-RARRES1-4 had the best interference efficiency, and it could significantly and extremely significantly down-regulate the expression of RARRES1 and BMPR-IB mRNA in follicular granulosa cells (P<0.05, P<0.01). In this study, the complete sequence of RARRES1 gene of Qianbei Ma goat was cloned and analyzed by PCR amplification and bioinformatics analysis, respectively. The recombinant plasmids pEGFP-N3-RARRES1, pGPH1/GFP/Neo-RARRES1 were successfully transfected into follicular granulosa cells of Qianbei Ma goat, and their expression changes were verified. The results showed that RARRES1 might have a promoting effect on fecundity in goats, which provided a basis for further exploring the regulation mechanism of RARRES1 gene on fecundity in goats.

The purpose of this experiment was to study the biological safety of sheep with melatonin synthase genes AANAT and ASMT (HIOMT) over-expressed in mammary gland. Based on the sheep models with AANAT and ASMT (HIOMT) over-expressed in mammary gland established in the previous study, we tracked the growth traits of positive transgenic sheep, analyzed the physiological and biochemical indexes of the blood and urine samples, and detected the intestinal microorganisms, melatonin levels in milk and milk composition. The results showed that:1) The body weight, body length, height and bust of positive transgenic sheep at 0, 6 and 12 months of age were not significantly different from those of ordinary sheep (P>0.05); 2) The total protein levels, albumin, globulin and cholesterol levels in blood of positive transgenic sheep as well as the number of various types of cells were not significantly different from those of ordinary sheep(P>0.05). Meantime, the levels of nitrite, protein, glucose and some other factors in urine were normal and not significantly different from those of ordinary sheep (P>0.05); 3) The sequencing results of microbiota showed that there was no significant difference in the composition of intestinal microbial communities and dominant bacteria between positive transgenic sheep and ordinary sheep(P>0.05); 4) The expression level of melatonin in blood of positive transgenic sheep was not significantly different from that of ordinary sheep (P>0.05), the level of melatonin at night was significantly higher than that at daytime (P<0.05); while the level of melatonin in milk of positive transgenic sheep was extremely significantly higher than that of ordinary sheep (P<0.01), the lactose content was significantly higher than that of ordinary sheep (P<0.05), in addition, the number of somatic cells of positive transgenic sheep was extremely significantly lower than that of ordinary sheep (P<0.01). In summary, the growth performance, many physiological and biochemical indexes and intestinal microbiota of positive transgenic sheep with AANAT and ASMT (HIOMT) over-expressed in the mammary gland were not significantly different compared with the ordinary sheep. Furthermore, the content of melatonin and lactose in milk of positive transgenic sheep were higher and the number of somatic cells was lower, which might be that melatonin in the mammary gland played an anti-inflammatory effect, reducing the number of somatic cells in the milk.

This study was conducted to investigate whether Tibetan goat ATP6(ATP synthase F0 subunit 6) and Cytb(cytochrome b) genes had potential adaptive mutations in high altitude environments. In this study, by comparing 261 ATP6 and Cytb sequences of Tibetan goats (n=157, from 8 populations across Tibetan plateau with altitude>3 500 m) and low-altitude goats (n=104, from 50 populations across Eurasia with altitude<1 000 m), the nucleotide composition, genetic diversity, haplotype network map, and the effect of gene mutations on protein domain function were compared and analyzed. Thirty-three and 67 single nucleotide polymorphisms (SNPs) sites were identified in ATP6 and Cytb, respectively, in which, both genes contained 6 missense mutations. ATP6 m.8102A>G (Ile → Met; P=0.000 6) and Cytb m.14794A>G (Thr → Ala; P=0.007 7) were unique missense mutations in low-altitude goat populations, which formed haplotypes H27 and Ha36 that were significantly negatively related to high altitude adaptation (H27, P=0.007 0; Ha36, P=0.012 3). Further analysis speculated that the two mutations hindered the normal function of the proton transmembrane and affected the transfer of protons or electrons in OXPHOS(oxidative phosphorylation), which led to the difference in respiration efficiency between low-altitude goats and Tibetan goats. Based on the correlation analysis of polymorphism to function of mitochondrial ATP6 and Cytb coding regions, the high-altitude adaption mechanism of Tibetan goats was preliminarily explored, which provided certain insights for related research on altitude adaptability.

The purpose of this study was to analyze the data of embryonic skeletal muscle proteomic in sheep, and provide a basis for revealing the mechanism of embryonic skeletal muscle development and growth in sheep. In previous proteomic study, the embryonic longissimus dorsi proteins were quantified by using tandem mass tag (TMT) in the adult Chinese merino ewes on the Day 85 (D85), 105 (D105) and 135 (D135) of gestation, and 3 comparable groups D85/D105, D105/D135 and D85/D135 were set up. The KEGG and parallel reaction monitoring (PRM) methods were used to analyze and verify the up-and down-regulation differential abundance proteins in sheep embryonic skeletal muscle in 3 comparable groups, and bioinformatics analysis was performed for the candidate proteins. The marker protein about muscle fibers maturation and differentiation of myosin-2 isoform X2 (MYH) was verified by PRM, the results demonstrated that the change trend of MYH was consistent with the trend of sheep embryonic muscle fiber maturation and differentiation. The up-regulation and down-regulation differential abundance proteins were analyzed by KEGG method, the results showed that the up-regulation differen-tial abundance proteins in D105/D85 comparable group were significantly enriched in insulin signaling pathway, and the down-regulation differential abundance proteins in D135/D105 and D135/D85 comparable groups were significantly enriched in DNA replication and protein digestion and absorption signaling pathways. The muscle development related proteins, cAMP-dependent protein kinase catalytic subunit alpha (PRKACA) and glucose transporter member 4 isoform X1 (GLUT4), were significantly enriched in insulin signaling pathway. The bioinformatics analysis of the two proteins revealed that the theoretical molecular weight of PRKACA and GLUT4 were 121.73 and 20.64 ku, respectively. The PRKACA contained 147 positively charged amino acid residues and 135 negatively charged amino acid residues, the theoretical isoelectric point was 8.81, the hydrophilic average coefficient was -0.408; and the GLUT4 contained 12 positively charged amino acid residues and 12 negatively charged amino acid residues, the theoretical isoelectric point was 6.54, the hydrophilic average coefficient was 0.811. There were no N-terminal glycosylation site and there was 45 phosphorylation sites in PRKACA; there was an N-terminal glycosylation site and 25 phosphorylation sites in GLUT4. The homology of PRKACA and CAMP-2 relies on the chimeric fusion crystal structure of protein kinase a was 85% in tertiary structure, the homology of the GLUT4 and human glucose transporter GLUT1 was 78% in tertiary structure. The results showed that the trend of MYH quantification verified by PRM was consistent with the trend of embryonic muscle fiber maturation and differentiation. PRKACA and GLUT4 were important candidate proteins, with abundant phosphorylation sites, involved in the regulating of insulin signaling pathway.

Since the discovery of angiotensin-converting enzyme 2 (ACE2),its pathophysiological functions,especially as functional receptors for coronaviruses such as SARS and COVID-19,have shown great potential.To clarify the gene sequence and structure of different species can provide a basis for the study of the mechanism of coronavirus infection. In this experiment,RT-PCR and Western blot were firstly used to detect the presence of ACE2 in different tissues of China Sheldrake duck.Then the homologous cloning and PCR technology were used to amplify the complete ORF sequence of the China Sheldrake duck ACE2 gene,and then TA cloned into pMD-19T.The vector was sequenced,and the obtained sequence was analyzed by bioinformatics. The expression of ACE2 gene and protein in heart,liver,lung,kidney and other tissues was confirmed.Gene cloning results showed that the full-length CDS sequence of the China Sheldrake duck ACE2 gene was 2 435 bp,encoding 805 amino acid residues,and its nucleotide sequence and amino acid sequence homology with human ACE2 were 66.2% and 66.4%,respectively,and on different branches of the evolutionary tree.Analysis of the 18 key amino acid residues related to the binding of the SARS virus S protein in humans found that except for the 330th and 353th amino acids, the rest were different from humans. Structural analysis revealed that the duck ACE2 was a type I transmembrane protein with multiple N-glycosylation sites.The study obtained the complete ORF sequence and related basic data of the ACE2 gene of China Sheldrake duck for the first time. The obtained sequence had been uploaded to GenBank and successfully included.The results provided a theoretical basis for the functional study of ACE2 on ducks.

The purpose of this study was to verify the results of SMAD1 gene up-regulation comparing single lamb group to multiple lambs group in transcriptome sequencing, and to explore the effect of SMAD1 gene on lambing traits of Qianbei Ma goats. In this experiment, healthy, 4-year-old single, multi-lamb Qianbei Ma goats ewes with 45 kg were selected as the research objects. qRT-PCR method was used to detect the expression of SMAD1 gene in uterus, fallopian tubes, pituitary, hypothalamus and ovarian tissues of Qianbei Ma goats ewes in the single and multi-lamb groups. CDS region of SMAD1 gene of Qianbei Ma goats was amplified, recombinant plasmid pEGFP-N3-SAMD1 was constructed and was transfected into ovarian granulosa cells by liposome transfection method. The SMAD1 overexpression test group and pEGFP-N3 empty vector control group were set up. By detecting the concentration of estradiol and progesterone in the culture medium, the effect of overexpressing SMAD1 gene on the proliferation and apoptosis of ovarian granulosa cells was detected by CCK-8 and Annexin V-FITC methods. qRT-PCR was used to detect the effect of overexpressing SMAD1 gene on the mRNA expressions of SAMD1, GnRHR, FSHR, BMP4 and TIMP3 genes to explore the influence of SMAD1 gene on ovarian granulosa cells. The results showed that SMAD1 gene was expressed in various tissues of Qianbei Ma goats. Comparison between the groups showed that the expression of SMAD1 gene in ovary and hypothalamus tissues were extremely significantly higher in the single lamb group than those in the multiple lambs group (P<0.01), in the fallopian tube tissues, it was significantly higher in the single lamb group than that in the multiple lambs group (P<0.05), and there were no significant difference in other tissues between the groups(P>0.05). The CDS sequence of SMAD1 gene was successfully cloned, no mutation site was found. The recombinant plasmid pEGFP-N3-SAMD1 was transfected into ovarian granulosa cells. The E2 content in the culture medium in transfection test group was significantly higher than that in the control group at 12 and 48 h (P<0.05), the content of P4 in transfection test group was significantly higher than that in the control group at 12 h (P<0.05); Overexpressing SMAD1 gene significantly promoted cell proli-feration at 24 h (P<0.05), and extremely significantly promoted cell proliferation at 48 and 72 h (P<0.01). It was found that overexpression of SMAD1 gene inhibited the apoptosis of ovarian granulosa cells, which was consistent with the result of SMAD1 gene promoting the proliferation of ovarian granulosa cells. However, the mRNA expressions of GnRHR, FSHR, BMP4, and TIMP3 were extremely significantly decreased after overexpressing SMAD1 gene(P<0.01). In summary, the overexpression of SMAD1 gene can increase the production of E2 and P4, help to maintain the estrus and pregnancy of Qianbei Ma goats, promote cell proliferation, inhibit cell apoptosis, and also significantly inhibit the expression of reproduction-related genes mRNA, and these reproduction-related genes can regulate the number, development and formation of follicles and ovulation. Therefore, the SMAD1 gene can suppress the expressions of reproduction-related genes, which results in a low conception rate in the single lamb group of Qianbei Ma goats, and further reduces the number of lambs. The result of this study provide a theoretical basis for studying the lambing traits related genes, and SMAD1 gene can be used as a candidate gene to explore goat lambing traits.

In order to guide the optimization of ovulation synchronization, the body temperature and physical activity of cows treated with ovulation synchronization and effect of different ovulation synchronization techniques were investigated. Eighteen Holstein cows (20-month-old) treated with ovulation synchronization (GnRH-PG-GnRH) and 17 Holstein cows (40-60 days postpartum) treated with ovulation pre-synchronization (PG-PG-GnRH-PG-GnRH) were selected, and their body temperature, physical activity and the estrus were monitored by automatic detection system. The results showed that the average vaginal temperature of cows treated with ovulation synchronization increased by (0.43±0.2)℃ and lasted for about (12.37±2.73) h, the average amount of physical activity increased (18.28±18.61) times and lasted for (11.00±1.68) h during estrus; the vaginal temperature decreased by (0.20±0.10)℃, and lasted for about (11.00±1.68) h during ovulation. Automatic estrus identification showed that, 7 cows treated with ovulation synchronization were in estrus and ovulation; all cows treated with ovulation pre-synchronization were in estrus and ovulation before GnRH treatment. Both ovulation synchronization techniques could change the course of cow sexual cycle and promote the synchronization of cow sexual cycle, but it was difficult to completely synchronize the sexual cycle of cows. Therefore, the scientific combination of ovulation synchronization-timed artificial insemination and estrus diagnosis technology can achieve better reproduction effects.

In this study we aimed to investigate the effects of intrauterine growth retardation (IUGR) on insulin levels and hepatic development in weaned piglets, and the repairing effect of dietary supplementation with 80 mg·kg^-1 dihydroartemisinin (DHA). Ten piglets with normal birth weight (NBW) and 20 IUGR piglets were selected and fed either a basal diet (NBW and IUGR groups, 10 piglets in each gorup) or the basal diet supplemented with 80 mg·kg^-1 DHA (IUGR-DHA group, 10 piglets) for 29 days from 21 to 49 d of age. Compared with NBW group, IUGR group significantly decreased the serum content of fasting glucose (FBG) and insulin-like growth factor-1 (IGF-1) and the hepatic weight (P<0.05), and significantly increased the content of fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR) values and hepatic index (P<0.05). At the same time, the morphology and structure of the hepatic tissue in IUGR weaned piglets were damaged, there was obvious vacuolation, and organelle structure was damaged, mitochondria and endoplasmic reticulum were significantly swollen, and the nucleus was seriously deformed. Compared with IUGR group, IUGR-DHA group significantly increased the hepatic weight and IGF-1 content in the serum (P<0.05). The serum content of FINS and HOMA-IR values were significantly decreased (P<0.05). In addition, the morphology and structure of liver tissue were significantly improved. Compared with NBW group, IUGR group significantly downregulated the relative mRNA expression of IGF-1, insulin receptor substrate 1 (IRS1), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT2) (P<0.05) in liver. Compared with IUGR group, IUGR-DHA group significantly upregulated the relative mRNA expression of IRS1 and AKT2 in the liver of IUGR weaned piglets (P<0.05). The results showed that DHA had a certain repairing effect on insulin resistance and hepatic development damage of weaned piglets caused by IUGR. This study provides a theoretical basis for the early treatment of IUGR and the promotion and application of DHA in livestock production.

This study was designed to investigate the effects of zearalenone (ZEA) on the antioxidative and inflammatory indexes and related genes expression of uterus and ovary in prepubertal gilts. A total of 48 Landrace×Yorkshire prepubertal gilts with similar birth parity, breeds and body weight ((23.20±0.68) kg)were randomly divided into 4 groups(12 replicates per group, 1 gilt per replicate). Prepubertal gilts in control group(CON group)were fed a basal diet and those in experimental groups(T1, T2 and T3 groups) were fed test diets supplemented with 200, 800 and 1 600 μg·kg^-1 ZEA in the basal diet, respectively. Pretest lasted for 7 d, and formal test lasted for 40 d. The results showed as follows:1)The serum T-AOC and SOD activity of prepubertal gilts in the T3 group were significantly lower than those in the CON group (P<0.05), and the MDA level was significantly higher than that in the CON group(P<0.05). 2)In the uterine tissues of prepubertal gilts in each group, T-AOC activity in group T2 and T3 significantly decreased than those in the CON group (P<0.05), and the SOD activity in the group T3 extremely significantly decreased than that in the CON group (P<0.01). In the ovarian tissues of prepubertal gilts in each group, the activity of T-AOC in the T3 group and the activity of GSH-Px and SOD in the T2 group were significantly lower than those in the CON group(P<0.05), and MDA level in the T3 group was significantly higher than that in the CON group(P<0.05). 3)The serum TNF-α and IL-4 levels of prepubertal gilts in the T2 group were significantly higher than those in the CON group(P<0.05)and IL-10 levels of prepubertal gilts in T1, T2 and T3 groups were significantly lower than those in the CON group(P<0.05). 4)In the uterine tissues of prepubertal gilts in each group, IL-1β level in the T2 group was significantly higher than that in the CON group(P<0.05), and IL-10 levels in T1 and T3 groups were significantly lower than those in the CON group(P<0.05). In the ovarian tissues of prepubertal gilts in each group, level of TNF-α in the T2 group was significantly higher than that in the CON group(P<0.05), and levels of IL-4 in T1, T2 and T3 groups were significantly higher than those in the CON group(P<0.05). 5)In the uterine tissues of prepubertal gilts in each group, the relative expression of Cu/Z_n-SOD mRNA in the T3 group was significantly lower than that in the CON group(P<0.05), and the relative expression of IL-1β mRNA in the T2 group was significantly higher than that in the CON group(P<0.05). 6)In the ovarian tissues of prepubertal gilts in each group, the relative expressions of GSH-Px mRNA in the T3 group was significantly lower than that in the CON group(P<0.05), and the relative expressions of IL-1β mRNA in the T2 group was significantly higher than that in the CON group(P<0.05). In conclusion, by inhibiting the expression and synthesis of SOD and GSH-Px, ZEA can reduce the antioxidant performance of uterus and ovary of prepubertalgilts and cause oxidative stress. ZEA can also cause inflammatory responses in the uterus and ovary of prepubertal gilts by promoting the expression and synthesis of TNF-α and IL-1β. Inhibition of IL-10 expression and synthesis inhibits the inflammatory response of the uterus and ovary.

To explore the effects and mechanism of high concentrate diet on hepatic and spleen tissues in dairy goats, 14 Guanzhong dairy goats at the early stage of lactation were randomly divided into control group (CG, n=6) and experimental group (EG, n=8). The CG was fed with a normal diet (concentrate:roughage=35:65), and the EG was fed with a high concentrate diet (concentrate:roughage=65:35). The trial lasted for 19 weeks. The milk from all goats was collected at 0, 1, 4, 8, 12, 15, 16, 18 and 19 weeks to analyze milk fat. Every Sunday from 6 to 18 weeks, all sheep feces were collected for pH determination. After autopsy at the end of the experiment, hepatic and spleen tissues were collected and tissue sections were made to observe the morphological changes of the hepatic and spleen tissues. The gene expression of inflammatory cytokines and key proteins of the TLR4 pathway in hepatic and spleen tissues were detected with the RT-qPCR method. The expression level of key proteins in the TLR4 pathway was analyzed using the Western blot method. The results showed that milk fat of EG decreased and was significantly different from those of CG at 16, 18 and 19 weeks (P<0.05). Average fecal pH of EG declined from the 9th week, and the difference was significant between EG and CG from 13 to 18 weeks (P<0.05). Hepatocyte was granule or vacuole degeneration and there was inflammatory cell infiltration around central vein of hepatic lobular. The spleen tissues' change was not obvious. The expression of pro-inflammatory factors IL-6 and TNF-β, as well as TLR4 and NF-κB2 genes were significantly increased in hepatic tissues (P<0.05). The expression of TNF-β, IL-10 and TLR4 genes were significantly increased, while the expression of NF-κB2 was significantly decreased in spleen tissues (P<0.05). p38, JNK, ERK and p65 proteins' expressions in hepatic tissues were not significantly changed, while the expressions of p38, ERK and p65 proteins in spleen tissues showed decreased trend. Those indicated that the decline of milk fat and fecal pH were induced by long-term feeding high-concentration diet. Upregulating of NF-κB gene expression led to the increased expression of IL-6 and TNF-β genes in the hepatic tissues of dairy goats, which caused inflammatory damage to the hepatic tissues, but had no obvious effect on the spleen tissues.

The aim of this study was to investigate the effects of diets with different nutrition levels on the growth performance and the mRNA expression of SLC7A7, SLC3A1 and SLC15A1 in intestinal tissues of 2-6 months old weaned Shaanbei white cashmere goats. Thirty-six healthy female Shaanbei white cashmere goats with age (60±1.60) d and body weight (10.73±1.03) kg were randomly divided into 4 groups and fed with 4 test diets, which digestive energy and crude protein were 85%, 100%, 115% and 130% of the standard diet, respectively. The nutrition level of standard diet formulated with reference to the NY/T816-2004. Goats were weighted at 120 and 180 d. One goat (a total of 12 goats) was slaughtered in each replicate. The upper middle part of the duodenum, the middle part of the jejunum and the ileum end tissue samples were collected. The mRNA expression levels of SLC7A7,SLC3A1 and SLC15A1 in the sample tissues were detected by real-time fluorescent quantitative PCR. The results showed that:1) In the first stage (60-120 d), the average daily gain of the 115% level group was significantly higher than those of other groups (P<0.05); In the second stage (121-180 d), the average daily gain of the 115% level group was significantly higher than 85% and 100% levels groups (P<0.05), but there was no significant difference compared to 130% level group (P>0.05). The dry matter intake of lambs in the 115% level group in the two stages were extremely significantly higher than those of other groups (P<0.01). The feed to gain ratio of 115% level group was significantly lower than those of 85% and 100% level groups (P<0.05). 2) With the same nutrition level, the mRNA expression levels of SLC7A7 and SLC15A1 were all in order of ileum>jejunum>duodenum; 3) The mRNA expression of SLC7A7, SLC3A1 and SLC15A1 in small intestines increased first and then decreased with the increase of dietary nutrient level, and the 115% level showed the highest expression. The mRNA expression of SLC7A7 and SLC15A1 in the 115% group was significantly higher than those of the the other three groups (P<0.05). The mRNA expression of SLC3A1 in the 115% group was significantly higher than those of the 85% and 130% groups (P<0.05). In this experiment, goats at age of 2-6 month-old fed the diet with 115% digestible energy and crude protein of the standard showed better growth performance, and the mRNA expression of SLC7A7, SLC3A1 and SLC15A1 in the small intestine was higher than those of the other groups.

This experiment was conducted to study the effects of diets with different neutral detergent fiber (NDF) levels on growth performance, nutrient digestion,digestive tract weight and rumen papilla development of lambs. Sixty healthy newborn female lambs ((3.23±0.20) kg initial body weight) were selected and randomly divided into 4 groups with 5 replicates each, 3 sheep each. The lambs were offered concentrates at 10 days of age, and NDF levels of concentrates were 12% (12NDF group), 16% (16NDF group), 20% (20NDF group) and 24% (24NDF group),respectively. During the trial period of 60 days, all lambs were breast-fed in the feeding test, and all lambs were not breast-fed in the digestion test. The results showed that:1) The body weight of the lambs increased extremely significantly with the increase of age (P<0.01); No significant difference in daily weight gain at all ages.With the increase of age, the dry matter intake (DMI) of the lambs increased extremely significantly (P<0.01). The DMI of the lambs among each group were not significantly different (P>0.05). 2) The dry matter (DM), organic matter (OM) intake and total energy (GE) intake of the 24NDF group were significantly higher than those of the 16NDF and 20NDF groups (P<0.05). The intake of crude protein (CP) of lambs in the 24NDF group was significantly higher than that of the 20NDF group (P<0.05); the intake of neutral detergent fiber (NDF) and acid detergent fiber (ADF) in the 24NDF group were extremely significantly higher than those of the 20NDF group (P<0.01), the 20NDF group was extremely significantly higher than those of 16NDF and 12NDF groups (P<0.01). The apparent digestibility of DM and OM in the 24NDF group were significantly lower than those of the 12NDF and 16NDF groups (P<0.05),the apparent digestibility of GE in the 24NDF group was significantly lower than that in the 16NDF group (P<0.05).The apparent digestibility of NDF and ADF in 20NDF and 24NDF groups were significantly or extremely significantly higher than those in 12NDF group (P<0.05 or P<0.01).The fecal energy of lambs in 24NDF group was significantly higher than those of 12NDF, 16NDF and 24NDF groups (P<0.05).There was no significant difference in CP apparent digestibility and digestive energy among the experimental groups (P>0.05).3) There was no significant difference in the weight of reticulum, omasum, abomasum,small intestine, jejunum and ileum in the lambs of each experimental group (P>0.05). The total weight of stomach in 12NDF group was signifcantly higher than those of 16NDF and 20NDF groups. The rumen weight of 12NDF group was significantly higher than that of 20NDF group (P<0.05). The proportion of rumen to live weight before slaughter in 12NDF group was significantly higher than those of 16NDF, 20NDF and 24NDF groups(P<0.05). The duodenum weight and the proportion of duodenum in live weight before slaughter in the 12NDF group was significantly higher than that in the 16NDF group (P<0.05). The other indicators were not significantly different(P>0.05). In conclusion, the optimal NDF level of diet for female lambs at the age of 0 to 60 days is 16%-20%.

To investigate the genetic characteristics of a porcine reproductive and respiratory syndrome virus (PRRSV) strain isolated in Henan Province in 2017, the complete genome was sequenced and further analyzed by bioinformatics. The results showed that the length of the genome is 15 054 bp, and the whole genome shared 84.5%, 93.4%, 84.2%, 83.9%, and 82.1% nucleotide similarities with VR-2332, NADC30, CH-1a, JXA1, and QYYZ, respectively. Phylogenetic analysis displayed that the isolate was clustered into the same branch with NADC30-like strains (lineage 1). For GP5 protein of the isolated strain, the N glycosylation sites were identified at 34, 44, and 51 sites. The linear epitope (37-45 aa) was conserved during NADC30-like strains while the other two linear epitopes (27-30 aa, 37-45 aa) have specific amino acid mutations. The Nsp2 of the isolate has 131 discontinuous amino acid deletion(323-433,483,504-522)which was a genetic marker of NADC30-like strains. Furthermore, the results of recombination analysis showed that the isolate was a mosaic virus between the major parent strain NADC30-like and minor parent strain JXA1-like. The recombinant region was in Nsp2 (1 340-2 082 nt). Our studies described the genetic characteristics of a NADC30-like strain and confirmed the recombinant events that happened in the Nsp2 region. It suggests that PRRSV undergoes genetic evolution through recombination and it is important to continuous monitoring of PRRSV prevalence and genetic variation.

The objective of this study was to isolate the exosomes of PK-15 cells infected with PCV2 and explore its role in lymphocyte infected with PCV2. The exosomes in the supernatant of PK-15 were extracted. We observed the morphology of the exosomes by transmission electron microscopy (TEM), measured the particle size of the exosomes by nanoparticle tracking analysis (NTA), and detected their molecular markers CD81 and TSG101 by Western blot. PKH67 labeled exosomes were used to detect the uptake of exosomes by cells. The PCV2-exosome genome was extracted and PCV2 Rep and Cap genes were detected. Indirect immunoinfluscent assay (IFA) was used to detect the localization of the PCV2 in PK-15 cells infected with PCV2-exosome. The infection rate of PCV2-exosome on lymphocyte was detected by absolute quantitative PCR. Lymphocyte proliferation were detected by using CCK-8 method. The apoptosis rate of lymphocytes was detected by Annexin V-FITC/PI. The results showed that exosomes had bilayer membrane vesicles with a diameter of 30-200 nm by using TEM and NTA. CD81 and TSG101, two exosomal protein markers were expressed in the purified exosomes. The results of the exosome uptake experiment showed that exosomes could be taken by PK-15 cells. The PCR results showed that PCV2-exosome genome contained PCV2 Rep and Cap genes. The IFA results showed that compared with PCV2 direct infection, PCV2 copies in lymphocytes infected with PCV2 exosomes were significantly higher (P<0.01). There was no significant difference between PCV2 virus and PCV2-exosome lysate on the lymphocyte infection ability. The lymphocyte proliferation was significantly inhibited by PCV2-exosome (P<0.01 or P<0.05). The lymphocyte apoptosis was significantly increased by PCV2-exosome. These findings suggest that the exosomes purified from PCV2 infected PK-15 cells contained viral genes, which can reduce cell proliferation and increase cell apoptosis in lymphocyte.

C-strain (also known as HCLV strain) of classical swine fever virus (CSFV) is a lapinized live attenuated vaccine against classical swine fever (CSF). However, C-strain lacks the capacity for the serological differentiation between infected and vaccinated animals (DIVA). The purpose of this study is to insert the gene encoding the enhanced green fluorescent protein (EGFP) into CSFV C-strain to construct a reporter virus, and to provide a virus tracing tool for basic research of C-strain, and also to provide a method for constructing C-strain marker vaccine. Here, we constructed and characterized two reporter viruses based on C-strain. One is the reporter virus rHCLV-EGFP, which expressing the enhanced green fluorescent protein (EGFP) fused in frame with the N^pro protein. The other one is the chimeric reporter virus rHCLV-N^pro(SM)-EGFP in which the N^pro gene is replaced with the counterpart of the highly virulent CSFV Shimen (SM) strain in the context of C-strain. The researchers identified the rescued reporter viruses by ELISA kit for CSFV antigen, observing the fluorescence directly and detecting the EGFP protein by Western blot, as well as evaluated the biological characteristics of these two reporter viruses in cells and rabbits. The antigens of the two reporter viruses were positive by ELISA. The reporter virus rHCLV-EGFP remained viable but not fluorescent, containing an intact EGFP gene but expressed as an N^pro-EGFP fusion protein with unexpected size. Interestingly, the EGFP fluorescence was restored in the chimeric reporter virus rHCLV-N^pro(SM)-EGFP. The experiment in cells showed that the two reporter viruses had similar growth characteristics to the parental virus and retained genetic stability. Furthermore, the animal experiment in rabbits revealed that rHCLV-N^pro(SM)-EGFP exhibited similar biological properties to the parental virus C-strain, while rHCLV-EGFP lost the ability to induce the fever response of C-strain in rabbits. In this study, the chimeric reporter virus rHCLV-N^pro (SM)-EGFP is able to be used as the reporter virus of C-strain. It can be used for virus tracing to study the replication process of the C-strain and the interaction between the virus and cells. The chimeric reporter virus also has the potential for a marker vaccine, which has the DIVA capability by detecting the anti-EGFP antibody.

This study aimed to establish a reliable indirect ELISA method for detecting PDCoV IgG antibody and to understand the prevalence of PDCoV in Sichuan. The prokaryotic expression system was used to express PDCoV membrane protein (M) as a detection antigen. By optimizing the reaction conditions, an indirect ELISA method named PDCoV-rM-ELISA based on PDCoV M protein was established, and 430 clinical samples were detected by this method. The recombinant expression E.coli strain BL21-pET-32a-M was successfully constructed, and the recombinant M protein was expressed after induction. Western blot assay showed that the recombinant protein had good reactivity. PDCoV-rM-ELISA method could specifically detect PDCoV antibody, and had no cross reaction with five common pathogen positive sera, suggesting good specificity; The sensitivity of this method was high. PDCoV-rM-ELISA was high reproducible with the coefficients of variation were less than 10%; Forty-eight clinic serums were detected using both PDCoV-rM-ELISA and PDCoV antibody detection kits. The results showed that the positive agreement rate was 88.46%, the negative agreement rate was 90.91%, and overall coincidence rate was 89.58%. The results of clinical samples detection showed that the average positive rate of PDCoV antibody was 11.86%, and the statistical analysis showed that the sows were 2.25 times more likely to be infected PDCoV than piglets and fattening pigs. An indirect ELISA was established by using the recombinant M protein of PDCoV express in E. coli BL21. PDCoV-rM-ELISA method had good specificity, reproducibility and stability, and could be used for the detection of PDCoV antibody in clinic.

Earlier in the laboratory, we screened the swine influenza virus (SIV) replication-related host factor ALG5(dolichyl-phosphate beta-glucosyltransferase) through CRISPR/Cas9 genome-wide knockout library in pig kidney cells (PK-15-GeCKO). Under this premise, the relationship between ALG5 and SIV replication was studied in depth. In this study, a plasmid LentiGuide-puro-ALG5 containing ALG5 gene guide RNA (gRNA) was constructed by CRISPR/Cas9, and the newborn pig tracheal epithelial (NPTr) cell line stably expressing Cas9 protein was infected by lentiviral packaging. ALG5 knockout monoclonal cell line was screened by limiting dilution method. The knockout level of ALG5 gene on NPTr cells was confirmed by target genome sequencing and Western blot. The cell viability of knockout cells and wild-type cells was verified by CCK-8 experiment. The effects of ALG5 knockout and overexpression on SIV replication were examined in combination with TCID₅₀ and Western blot experiments. At the same time, the expression of endogenous ALG5 protein was detected after SIV infected NPTr cells. Finally, the effect of ALG5 knockout on replication of swine flu strain F26 was tested. NPTr monoclonal cell line that knocked out ALG5 (ΔALG5) was obtained, and there was no significant difference in cell activity between ALG5 knockout cells and wild-type cells. ALG5 gene knockout significantly inhibited SIV proliferation, and overexpression promoted SIV proliferation. Under viral infection conditions, the protein expression of ALG5 was up-regulated as the virus proliferated. ALG5 gene knockout can also inhibit the replication of swine flu strain F26, without strain specificity. This study indicates that ALG5 is an important host factor affecting the replication process of swine influenza virus. This study lays the foundation for studying the replication and pathogenic mechanism of swine influenza virus.

Bovine norovirus (BNoV) is an emerging causative agent of calf diarrhea in China, the aim of the study was to establish a real-time PCR assay for detecting BNoV. One pair of primers was designed based on RNA-dependent RNA polymerase (RdRp) gene of BNoV. The EvaGreen real-time PCR assay was successfully developed after the optimization of amplification conditions. The test results showed that the Ct value showed a good linear relationship with the standard in the range of 2.24×10²-2.24×10⁸copies·μL^-1 and the correlation coefficient R²=0.997, and the amplification efficiency was 98.44%. There is no specific amplification of other common calf diarrhea pathogens, only BNoV were positive. The detection limit of the method was 22.4 copies·μL^-1 for BNoV. The inter-assay and the intra-assay coefficient of variation were both less than 2%, indicating a good repeatability. 221 clinical samples that collected from the diarrheic calves in Henan Province during September 2017 to May 2019 were detected using this real-time PCR assay, and the BNoV detection rate was 11.31% (25/221), the farms positive rate was 92.86% (13/14). These results indicated that the real-time PCR assay has good sensitivity, specificity and repeatability, which can be provide an effective means for detection and epidemiological investigation of BNoV.

The aim of this study was to determine whether Trichomonas gallinae (T. gallinae) secrete exosomes and investigate the protein composition of exosomes derived from T. gallinae. T. gallinae was isolated from upper digestive tract of infected pigeons and identified by microscopy and sequencing. Exosomes were extracted from serum absent culture medium, and further verified by transmission electron microscopy, Western blot and nanoparticle tracking analysis (NTA). Label-free was used to identify the proteins of exosomes. Results showed that the parasite had T. gallinae morphology, and sequence alignment of ITS1/5.8S/ITS2 gene showed an identity of 98%. The isolated vesicles presented a typical cup-shaped morphology; NTA results showed that particle size concentrated on 125.1 nm, percentage of peak diameter at 132.3 nm was 99.3%; proteins such as CD63 and TSG101 were conspicuously detected, suggesting that the extracts were exosomes. Mass spectrometry results showed that enolase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate mutase and elongation factor were the highly expressed proteins. GO pathways showed that proteins of exosomes mainly enriched into cellular components such as intracellular and cytoplasmic, played GTP binding and GTPase activity function, and participated in biological process of small GTPase mediated signal transduction and glycolytic process. KEGG enrichment analysis showed that proteins of exosomes were enriched in pathways as metabolic pathways, glycolysis/gluconeogenesis, biosynthesis of antibiotics and secondary metabolites. These results indicated that T. gallinae can secrete exosomes and proteins of exosomes might play roles in energy metabolism, signal transduction and biosynthesis.

Previously 57 Escherichia coli phages were isolated and identified from the intestines of healthy pigs, and 12 of them were found to lyse enterotoxigenic Escherichia coli (ETEC). To gain a deep insight of the protective effect of the 12 coliphages on the animal hosts, we determined the host range and bacteriostatic efficacy of the 12 coliphages. 89 E.coli strains (including five ETEC strains) isolated from pig feces previously were used as indicator strains for host spectrum determination. The host range of coliphages was conducted by the spot test method, double-layer agar plate method and liqud LB culture. Bacteriostatic ability of coliphages on ETEC was evaluated by turbidity measurement. The results showed that all the 12 coliphages could lyse one or more ETEC strains, however, none could produce progeny by infecting ETEC. Nevertheless, seven coliphages (S143_1, S143_2, S144_2, S35, S86_1, L86 and S172) could significantly inhibit the growth of ETEC at MOI of 10 or 100. TEM observation revealed that 8 of the 12 coliphages were T4-like coliphages and the other four belonged to Myoviridae. These results suggest that the coliphages isolated from porcine gut which could not infect ETEC but lyse ETEC still inhibit growth of ETEC.

Pseudomonas aeruginosa is the main pathogen causing rotten eggs during incubation, we attempted to control it with lytic phage. In this work, a strain of virulent phage MH12-Q was sifted from the river water sample by using Pseudomonas aeruginosa PA-MH12 isolated from the rotten eggs. The biological characteristics of the phage were studied and its prevention and control effect on Pseudomonas aeruginosa induced rotten egg were tested. The phage MH12-Q was sequenced and identified as Myoviridae. The titer of phage MH12-Q can reach 10¹³pfu·mL^-1, the complete lysis concentration (CLC) of phage MH12-Q is 10¹⁰pfu·mL^-1, and the optimal growth temperature is 37℃. The optimum pH is 6-8 for the stability of the phage and its titer can maintain a high level at pH 4-10. Its optimal multiplicity of infection (OMOI) is 0.001, the incubation period of infected hosts is 30 min, and the outbreak period is 70 min. The phage with the biological properties above can reduce the infection rate of high concentration Pseudomonas aeruginosa from 70% to 40%. The biological properties of Pseudomonas aeruginosa phage MH12-Q are suitable for production and application, which has a good prevention and control effect on rotten eggs in hatchery and can improve the survival rate of chicken embryos infected with Pseudomonas aeruginosa.

This study was conducted to investigate the distribution and expression of platelet-derived growth factor receptor-β (PDGFR-β)in yak lungs, and to explore the adaptation mechanism of yak lungs to high altitude hypoxia environment. Immunohistochemistry, Western-blot technology and qRT-PCR were used to study the exact distribution and expression of PDGFR-β in the lungs of newborn, 1-year-old, 3-year-old, and 6-year-old yak. The results showed that PDGFR-β was mainly distributed in the bronchial and epithelial cells of the lung, pulmonary vascular endothelial cells, and smooth muscle cells of the bronchial wall, and its expression was strong. A small amount PDGFR-β was distributed in the smooth muscle cells of the bronchial wall and alveolar septal cells, and its expression was weak.PDGFR-β mRNA and protein were expressed at different levels in the lungs of yak at different ages. In terms of protein level, the newborn group was extremely significantly higher than those of the other 3 groups (P<0.01); in the 1, 3, and 6 years old groups, PDGFR-β expressions were not significantly different when compared with each other (P>0.05). In the mRNA expression level, the expression in the newborn group was the strongest and extremely significantly higher than those of the other 3 groups (P<0.01); PDGFR-β expression in the 1-year-old group was extremely significantly higher than the 3-year-old group and 6-year-old group (P<0.01); the expression difference was significant between the 3-year-old group and the 6-year-old group (P<0.05). The results indicated that PDGFR-β played an important role in the development of airways and pulmonary arteries and in the formation of adaptive structures in yaks at different ages, and this was most obvious in the newborn to 1 year old period.

This study aimed to understand the epidemiological situation of swine influenza virus (SIV) and analyze its evolutionary and genomic characterization in Henan province. In April 2018, 150 nasal swabs were collected from a pig herd with suspected influenza clinical signs to isolate the virus. The isolated virus was sequenced and analyzed. The pathogenicity was evaluated by infecting BALB/c mice. A strain of H1N1 subtype SIV was isolated and named of A/swine/Henan/NY20/2018(H1N1). Phylogenetic analysis results demonstrated that HA and NA genes of the isolated virus belong to the Eurasian avian H1N1 lineage, PB2, PB1, PA, NP and M genes belonged to the pdm/09 H1N1 lineage, and NS gene belonged to the classical swine H1N1 lineage. The HA protein cleavage site of the isolated is PSIQSR↓GL, which accords with the molecular characteristics of low pathogenic avian influenza virus. It can effectively replicate in the lungs and turbinates of mice and can cause pathological changes in lung tissue. A triple reassortant H1N1 subtype virus was isolated in this study, which has a certain pathogenicity to mice, suggesting that further monitoring of SIV should be strengthened.