Transportation is a major stressor in swine production, inducing endocrine imbalance, oxidative stress, and immune dysfunction. This study investigated the effects of Astragalus polysaccharides (APS) on transport-induced stress in piglets. Piglets were assigned to a control group (CON), a transport stress group (TS), and two APS-supplemented groups receiving low (APS-L) or high (APS-H) doses. Blood samples were collected on 1, 3, and 7 days post-transportation to assess stress hormones, oxidative status, and inflammatory responses. Compared with the CON group, piglets of TS group significantly elevated norepinephrine (NE) and epinephrine (EPI) levels, increased reactive oxygen species (ROS), reduced glutathione peroxidase (GSH-Px) activity, and upregulated pro-inflammatory cytokines (TNF-α and IL-6). APS supplementation attenuated these transport-induced changes. At 1 day after transportation, piglets in both APS-L and APS-H groups exhibited lower NE, EPI, ROS, TNF-α, and IL-6 levels and higher GSH-Px activity than those in the TS group. The APS-H group showed greater protective effects than the APS-L group. By 3 and 7 days, oxidative and inflammatory parameters in APS-treated piglets gradually normalized, with the APS-H group showing values more comparable to those of the CON group. These results indicate that APS alleviates transport-induced stress by modulating the endocrine response, reducing oxidative stress, and attenuating inflammatory responses. APS supplementation may serve as a nutritional strategy to improve the health and welfare of piglets during transportation.

Sow colostrum is rich in lactoferrin (LF), which can be orally administered to protect piglets from porcine epidemic diarrhea virus (PEDV) infection, thereby reducing piglet mortality. Previous study has shown that sows fed with recombinant B. subtilis expressing 4,4-diaponeurosporene (B.S-Dia) have significantly higher LF levels in their colostrum compared to sows fed with B. subtilis. This suggests that 4,4-diaponeurosporene (DNP) produced by B.S-Dia may influence LF content in sow colostrum. In this study, we first extracted DNP expressed by the recombinant probiotic using acetone-hexane extraction. Flow cytometry, RT-qPCR, and ELISA analyses demonstrated that DNP promoted dendritic cell (DCs) maturation and increased the expression of IL-1β and IL-27. We then established a method for isolating neutrophils from sow colostrum and set up a co-culture system of neutrophils and DCs to investigate factors regulating LF secretion. The results indicated that DCs secretions enhanced LF expression in neutrophils. Finally, the application of IL-27 inhibitors confirmed that IL-27 produced by DCs upregulates LF secretion in neutrophils. These findings elucidate the mechanism by which DNP promotes LF production in colostrum and provide a theoretical foundation for using B.S-Dia to prevent and control PEDV infection in piglets.

The colostrum of sows contains a large number of lymphocytes originating from breast tissue. However, the mechanism of lymphocyte migration across the mammary epithelium remains unclear. Hormonal fluctuations during pregnancy are known to influence lymphocyte homing to the breast. In this study, we established a mammary epithelial cells (MECs)/T lymphocytes co-culture system to investigate the effects of colostral hormones on lymphocyte migration. Electron microscope and fluorescence-activated cell sorting results showed that the proportion of T lymphocytes in colostrum was about 9 %. Subsequently, the hormone concentration and number of T lymphocytes in colostrum were tested. The results revealed that there is a high content of prolactin and estrogen in colostrum, and the content of prolactin and estrogen is positively correlated with the number of T lymphocytes in colostrum. Furthermore, T lymphocytes were added to the upper ventricle in Transwell insert, and the estrogen and prolactin were added to the lower ventricle. The results showed that prolactin significantly promoted the T lymphocyte migration across the mammary epithelium, while estrogen had no effect. Finally, our study found that prolactin can promote the mammary epithelial cell to secrete CCL2 by Real-time quantitative PCR (RT-qPCR) and Enzyme-linked immunosorbent assay (ELISA). In order to further explore the transepithelial migration, a neutralizing antibody was applied to block CCL2. Results revealed that the migration of T lymphocytes across the breast epithelium was significantly inhibited after blocking CCL2. The above results showed that prolactin can promote the secretion of CCL2 by breast epithelial cells, and regulate the lymphocyte migration across the mammary epithelium into colostrum, and thereby may further improve lactogenic immunity in sows.

Neonatal piglets infected with Porcine Epidemic Diarrhea Virus (PEDV) experience a mortality rate of up to 90%, resulting in significant economic losses to the swine industry in China. Current strategies using specific antibodies in sow milk to prevent Porcine Epidemic Diarrhea (PED) in these piglets through specific antibodies in sow milk is unsatisfactory. Preliminary studies have shown limited success. Emerging evidence suggests that general immune factors in sow milk provide protection to neonatal piglets, particularly, lactoferrin, play a crucial role in protecting piglets by inhibiting PEDV replication. However, the precise mechanism by which lactoferrin exerts its antiviral effects remains unclear. This study sought to clarify these mechanisms through both in vitro and in vivo approaches, proposing that higher concentrations of lactoferrin lead to greater antiviral activity. It was hypothesized that lactoferrin can impede PEDV by blocking its binding to heparan sulfate proteoglycans (HSPG) on the surface of target cells, and molecular docking experiments was conducted to identify the binding sites between lactoferrin and HSPG. Additionally, the findings indicated that lactoferrin can effectively trigger the maturation of porcine dendritic cells and boosts their antigen-presenting functions, thereby improving intestinal mucosal immunity in neonatal piglets against PEDV. Overall, these findings aid to elucidate the antiviral actions and mechanisms of lactoferrin in sow colostrum, offering new insights for the effective prevention and control of PED in neonatal piglets.

Diarrhea in piglets causes intestinal inflammation and epithelial damage. Weaned piglets fed with Bacillus subtilis (B.S) have enhanced intestinal mucosal immunity and reduces diarrhea in piglets. However, the immune system of newborn piglets is immature, and B.S cannot effectively activate the intestinal mucosal reaction when given directly. This research explored the impact of the maternal supplementation of B.S-Dia during the final 35 days of gestation on piglet intestinal development and mucosal immunity. The results demonstrated that B.S-Dia administration significantly increased the body weight, jejunal villus height, and crypt depth in the piglets. In addition, B.S-Dia also significantly increased the proliferative activity of intestinal epithelial cells, as evidenced by proliferating cell nuclear antigen (PCNA) staining and the elevated mRNA expression of the proliferation-related gene (c-Myc). Furthermore, B.S-Dia supplementation also reinforced the intestinal mucosal barrier by increasing goblet cell numbers and upregulating the mRNA expression of antimicrobial peptides, such as Muc2 and Lyz-1. Finally, elevated levels of IL-4 and IFN-γ, along with an increased abundance of CD3+ T cells, revealed that the intestinal mucosal immunity of piglets was improved after B.S-Dia administration. Our study indicates that feeding B.S-Dia to sow spromotes intestinal development and improves intestinal mucosal immunity in piglets.

Schisandra chinensis polysaccharides (SCP) are isolated from Schisandra chinensis with a mean molecular weight of 32.401 kDa. Previous studies have demonstrated that Schisandra chinensis extract effectively prevented colitis and modulated gut microbiota in dextran sulfate sodium (DSS)-induced mice. In this study, 3% DSS was used to induce acute ulcerative colitis in mice, and then the SCP with different doses (50 mg/kg, 100 mg/kg, 200 mg/kg) and 5-aminosalicylic acid (5-ASA) were used to treat mice by oral gavage for one week. Here, we report for the first time that SCP presents outstanding protective effects on colon damage induced by DSS in mice. Firstly, SCP decreased oxidative stress, downregulated the expression of inflammatory factors, and inhibited the TLR4/p65/I kappa B alpha pathway in the colon. Then, SCP reversed DSS-induced intestinal barrier damage by improving the tight junction proteins. Moreover, SCP improved the levels of SCFA in the intestine of mice. Furthermore, SCP increased the diversity of gut microbiota and the abundance of beneficial bacteria, especially Lactobacillus and Akkermansia. Our results indicated that the protective effect of SCP on colitis caused by DSS may be due to inhibiting the TLR4/p65/I kappa B alpha pathway and the regulation of gut microbiota.

Rotavirus (RV), as the main cause of diarrhea in children under 5 years, contributes to various childhood diseases. Valeriana jatamansi Jones is a traditional Chinese herb and possesses antiviral effects. In this study we investigated the potential mechanisms of V. jatamansi Jones in RV-induced diarrhea. MTT assay was performed to evaluate cell proliferation and the diarrhea mice model was constructed using SA11 infection. Mice were administered V. jatamansi Jones and ribavirin. Diarrhea score was used to evaluate the treatment effect. The enzyme-linked immunosorbent assay was performed to detect the level of cytokines. Western blot and quantitative reverse transcription-PCR were used to determine protein and mRNA levels, respectively. Hematoxylin-eosin staining was applied to detect the pathological change of the small intestine. TdT-mediated dUTP nick-end labeling was conducted to determine the apoptosis rate. The results showed V. jatamansi Jones promoted MA104 proliferation. V. jatamansi Jones downregulated phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in protein level, which was consistent with the immunohistochemistry results. Moreover, V. jatamansi Jones combined with ribavirin regulated interleukin-1β (IL-1β), interferon γ, IL-6, tumor necrosis factor α, and IL-10, and suppressed secretory immunoglobulin A secretion to remove viruses and inhibit dehydration. V. jatamansi Jones + ribavirin facilitated the apoptosis of small intestine cells. In conclusion, V. jatamansi Jones may inhibit RV-induced diarrhea through PI3K/AKT signaling pathway, and could therefore be a potential therapy for diarrhea.

【Objective】 This study was aimed to explore the effect of Agrocybe aegerita polysaccharid（AAP）on immunomodulatory of splenic lymphocytes in mice,in order to provide a basis for elucidating the action mechanism and developing the clinical application of AAP．【Method】The splenic lymphocytes in mice were taken as the subjects,the MTT method was used to determine the safe concentration of AAP on lymphocytes,and the safe concentrations were selected for subsequent experiments.MTT assay was also used to detect the effect of different concentrations of AAP on the proliferation of splenic lymphocytes in mice.Three control groups of cell,lipopolysaccharide（LPS）and plant hemagglutinin（PHA）were also set at the same time to screen out AAP with the stronger proliferation ability on splenic lymphocytes in mice,flow cytometry was used to detect the effect of AAP on the values of CD4 + /CD3 + and CD8 + /CD3 + of T lymphocyte subtypes.ELISA method was used to determine the effect of AAP on the contents of interleukin-2（IL-2）,IL-4,tumor necrosis factor alpha（TNF-α）and gamma interferon（IFN-γ）in the supernatant of lymphocytes．【Result】AAP concentration lower than 1 000μg/mL had not any toxic effect on splenic lymphocytes in mice.Compared with control group,different doses of AAP could extremely significantly cooperate with LPS and PHA to stimulate lymphocyte proliferation（P＜0.01）,125and 250μg/mL AAP could extremely significantly increase the values of CD4 + /CD3 + and CD8 + /CD3 + （P＜0.01）,and significantly or extremely significantly increase the contents of IL-2,IL-4,TNF-αand IFN-γsecreted by lymphocytes（P＜0.05or P＜0.01）．【Conclusion】 AAP could cooperate with LPS and PHA to stimulate lymphocyte proliferation in spleen of mice,increase the values of CD4 + /CD3 + and CD8 + /CD3 + on the surface of lymphocytes and the ability of lymphocytes to release cytokines,thus improving the immune function of the body.

兽药残留检测中,样品基体涉及各种动物组织以及动物性加工食品,由于样品基体本身所存在的差异性使得在分析过程中不能简单地使用纯度标准物质去作对照,而需要使用更加接近实际样品的基体标准物质来满足检测需要.近些年,国内标准物质研发速度较快,数量也在稳步增长,但是动物源性食品相关的兽药基体标准物质数量却很少,远不能满足检测需求,被国家标准物质资源共享平台收录的仅40多种,存在着较大的缺口.本文重点综述了国内外动物源性食品相关兽药基体标准物质的研究进展和关键技术,分析了我国当前存在的问题短板和出现这些问题的原因,以期后续能为相关的研究提供一些参考.

Objective: This study aimed to provide the basis of quantity traceability for the analysis of amoxicillin residues in milk. At the same time, it can also provide quantity standards for ensuring the effective implementation of amoxicillin monitoring in milk and combating the abuse of amoxicillin in the country and regions. Methods: Two healthy middle-class dairy cows were injected with amoxicillin intramuscularly. After the natural metabolism of the drug for a period of time, the milk containing the target drug amoxicillin was collected and made into milk matrix reference material containing amoxicillin by vacuum freeze-drying technology. The uniformity test, stability test, fixed value and uncertainty evaluation of the prepared milk lyophilized powder matrix reference material were carried out. Results: In the uniformity test, the F value obtained is ＜F 0.05 , The results showed that the preparation of the standard milk powder was homogeneous; In the stability test, the short-term stability test（25, 4,-20 ℃; 2 weeks） and long-term stability test（4,-20 ℃; 6 months） showed good stability; The reference materials were jointly calibrated by 8 laboratories, and the final calibration result was 4.05 μg/kg, the total uncertainty being 0.2 μg/kg. Conclusion: The prepared amoxicillin matrix reference material in milk freeze-dried powder has good uniformity and stability, which can be used as a carrier to trace the amount of amoxicillin in milk, and can be widely used in the methodology verification and quality control of amoxicillin detection in milk.

In this experiment, the polysaccharide was extracted from Pueraria lobata (Willd.) Ohwi, and its structural characteristics and bioactivity were investigated. The results showed that Pueraria lobata polysaccharide (PLP) was composed of fucose, arabinose, galactose, glucose, xylose, mannose in a molar proportion of 0.09:1.25:2.19:95.74:0.43:0.30 with a number molar masses (Mn) weight of 14.463 kDa. Besides, FT-IR, Methylation, and NMR analysis revealed that PLP were mainly composed of the main chain →4)-α-Glcp (1→ and →4,6)-α-Glcp (1→, and the branched chain α-Glcp (1→. In vitro experiment, the results showed that PLP could stimulate the expression of surface molecules on RAW264.7 and (T and B) lymphocytes proliferation, simultaneously to stimulate their cytokines secretion. In vivo experiment, the immune organ index, cytokine content, and T lymphocyte subtype in cyclophosphamide-induced immunosuppressed mice could be improved by PLP. These data proved that PLP could be used as a useful immunomodulator to enhance the immune activity of RAW264.7, T, and B cells and improve the immune function of cyclophosphamide-treated mice.

旨在研究硒化修饰提高大蒜多糖的体外抗新城疫病毒(NDV)活性并筛选出体外抗NDV活性较好的硒化大蒜多糖.依次通过水提醇沉法、Sevage法去蛋白得到大蒜多糖(GPS),并采用硝酸-亚硒酸钠法对大蒜多糖进行硒化修饰,得到9个硒化大蒜多糖8GPS1～sGPS9.以鸡胚成纤维细胞(CEF)为作用对象,采用MTT法通过先加硒化大蒜多糖,或先加NDV,或两者同时加入3种方式比较9个硒化大蒜多糖抗NDV侵入、复制、直接抑制NDV的作用,同时设置细胞对照(CC)和病毒对照(VC),并计算病毒抑制率;筛选出活性较好的sGPS3、sGPS5、sGPS6测定其对NDV感染CEF后细胞凋亡、细胞周期的影响以及其对鸡胚的存活数的影响,同时设置GPS对照、VC对照、空白对照(BC).结果表明,GPS5、sGPS6组的细胞凋亡率、处于S期的细胞百分数显著低于VC组和GPS组(P＜0.05);sGPS6组36 h、48 h的细胞凋亡率显著小于sGPS3、GPS5组(P＜0.05),处于S期的细胞百分数小于sGPS3、sGPS5组,sGPS3组在36h、sGPS5组在24 h和36 h、sGPS6组在3个时间点的鸡胚死亡数均显著小于GPS组(P＜0.05),sGPS6组在3个时间点的鸡胚死亡数均为最低.综上,硒化修饰能提高大蒜多糖抗NDV的活性,其机制可能与阻止NDV侵入、抑制NDV诱导CEF凋亡有关.其中sGPS6的活性最强,可作为抗NDV的候选药.

一个专业的信息化教学有利于深化产教融合,而一个专业的信息化必然是由各门专业课程的信息化逐步完善起来的,高职《中兽医学》课程是高职动物医学专业的核心课程,也是全国执业兽医资格考试的必考科目.但传统的《中兽医学》教学模式不利于培养服务于产业的技术技能型人才.因此,本文围绕我校高职《中兽医学》课程信息化的最初设计、建设过程、具体应用、评价与反馈、后期完善等几个方面进行介绍,以期通过该课程的信息化设计与应用改变《中兽医学》传统教学模式,之后将此经验分享到高职院校动物医学专业的其他课程的教学实践中,更好地服务于兽医诊疗行业的技术技能型人才,从而为深化产教融合提供支撑.

This study investigated the structure of acidic Pueraria lobata polysaccharide (a-PLP) and its bioactive effects on intestinal function in cyclophosphamide (CY)-treated mice. The structure of a-PLP was preliminarily analyzed, and the results showed that it is composed of fucose, arabinose, rhamnose, galactose, glucose, xylose, mannose, galacturonic acid, and glucuronic acid in a molar proportion of 2.54:16.52: 6.14: 16.60: 4.05: 4.75: 0.48: 47.44: 1.47 with a weight average molecular weight of 22.675 kDa. In addition, the methylation analysis suggested that 4-Gal(p)-UA may be the main backbone of a-PLP. Furthermore, a-PLP (1.2 g/kg, 0.8 g/kg, and 0.4 g/kg) was administered orally for the treatment of CY-treated mice. The results showed that a-PLP could remarkably relieved weight loss and intestinal villous atrophy in CY-treated mice. Meanwhile, the secretion levels of sIgA, β-defensin, cytokines, Mucin-2, and tight junction proteins increased significantly. Moreover, the ratio of T (CD4+ and CD8+) cells in the Peyer's patches and mesenteric lymph nodes also increased remarkably, along with the number of goblet cells. Furthermore, a-PLP decreased the levels of diamino oxidase and malondialdehyde, but up-regulated the activity of superoxide dismutase. In summary, a-PLP exhibited great benefits by attenuating CY side effects, opening a potential avenue to effectively treat cancer and reduce the suffering of chemotherapy patients.

[目的]研究硒化大蒜多糖(sGPS)对小鼠腹腔巨噬细胞功能的影响,以期为硒化大蒜多糖的作用挖掘和临床应用提供依据.[方法]依次通过分离、纯化得到大蒜多糖(GPS),并经硝酸-亚硒酸钠硒化修饰得到sGPS3、GPS5和sGPS6.以小鼠腹腔巨噬细胞为研究对象,用6.25、12.5、25、50、100 μg/mL sGPS3、GPS5、sGPS6及10 μg/mL脂多糖(LPS组)处理小鼠腹腔巨噬细胞48 h,同时设置不加药物的细胞为对照组,用中性红法测定其吞噬功能,CCK-8法测定其增殖能力,筛选出活性最好的硒化大蒜多糖;然后用ELISA法检测活性最好的硒化大蒜多糖对巨噬细胞上清液中一氧化氮(NO)、肿瘤坏死因子-α(TNF-α)、干扰素-γ(IFN-γ)、白介素-1β(IL-1β)、白介素-6(IL-6)、白介素-12(IL-12)含量的影响;再通过小鼠碳廓清实验、小鼠腹腔巨噬细胞吞噬鸡红细胞实验测定空白对照组(生理盐水)及高(2 mg/mL)、中(1 mg/mL)、低(0.5 mg/mL)剂量活性最好的硒化大蒜多糖的吞噬指数与吞噬百分率.[结果]除6.25 μg/mL sGPS3外,6.25、12.5、25、50和100 μg/mL sGPS3、sGPS5、sGPS6组小鼠腹腔巨噬细胞的A490nm值均显著高于对照组(P＜0.05),其中,50和100μg/mL sGPS6组的A490nm值显著高于LPS组(P＜0.05),因此选用sGPS6进行后续试验;12.5、25、50和100 μg/mL sGPS6组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-1β、IL-6含量均显著高于对照组,25、50和100 μg/mL sGPS6组小鼠腹腔巨噬细胞上清中IFN-γ、IL-12显著高于对照组(P＜0.05),其中100 μg/mL sGPS6组小鼠腹腔巨噬细胞上清中NO、TNF-α、IL-12显著高于LPS组(P＜0.05).2和1 mg/mL sGPS6组的吞噬指数和吞噬率均显著高于空白对照组(P＜0.05).[结论]硒化大蒜多糖能增强小鼠腹腔巨噬细胞的吞噬功能和增殖能力,促进细胞因子TNF-α、IL-1β和IL-6的分泌.

为了给羊奶中恩诺沙星的残留检测提供实物基体标准物质,并满足在恩诺沙星残留检测过程中质量控制和方法学验证的需求,需要研制一批羊奶冻干粉中恩诺沙星基体标准物质.选取3只健康的泌乳期奶山羊,肌肉注射恩诺沙星后收集羊奶,利用冷冻干燥技术制得一批羊奶冻干粉恩诺沙星基体标准物质.对制备好的羊奶冻干粉恩诺沙星基体标准物质进行均匀性、稳定性评价、定值、不确定度评估.在均匀性评估中,获得的F值1.92＜F0.05(14,30),在稳定性评估中,分别进行了短期稳定性实验(4、-20、25℃,2周)和长期稳定性实验(-20、-80℃,12个月).数据表明,羊奶冻干粉恩诺沙星基体标准物质的均匀性、稳定性良好,8家实验室联合定值,制备的标准物质最终定值结果为101.08 μg/kg,总不确定度为2.7 μg/kg.

黄炎培是中国杰出的教育家和社会活动家,1916年9月在江苏省教育会内设立了职业教育研究会,1917年5月在上海成立了中国近代史上第一个研究、试验、推行职业教育的全国性团体中华职业教育社,提出了职业教育的教学原则:"手脑并用"、"做学合一"、"理论与实际并行"、"知识与技能并重".《中华人民共和国国民经济和社会发展第十四个五年规划和2035年远景目标纲要》明确提出增强职业技术教育适应性,实施现代职业技术教育质量提升计划.中国特色高水平高职学校和专业建设计划、实施职业教育提质培优行动计划(2020～2023年)标志着我国职业教育从规模发展转向提质培优、增值赋能、以质图强的职业教育现代化.时代虽然在发展,但黄炎培职业教育思想仍然对当前提升职业教育适应性的思考和实践具有重要指导作用.

旨在探究自体血小板凝胶(APRG)对巴马香猪人工创面愈合的影响,为APRG应用于创面愈合提供理论依据和实践支持.4只体重15～20 kg小香猪随机分为对照组和APRG组,每组2只,建立巴马香猪皮肤创伤模型,每只6个创面.于术后第7、14、21和28天观察创面大体愈合情况,进行血液中白细胞数(WBC)、红细胞数(RBC)、血红蛋白(HGB)含量、红细胞压积(HCT)、血小板数(PLT),以及血清中白介素8(IL-8)和白介素10(IL-10)含量测定;于术后第10和20天,无菌采集创面组织,进行HE组织观察和组织中羟脯氨酸(Hyp)含量测定.结果显示:与对照组相比,APRG组创面愈合速度和效果较好;HE染色结果显示APRG可以调节炎性细胞浸润,促进表皮和真皮恢复;APRG也能显著降低创面愈合过程中血液中白细胞数和血清IL-8含量(P＜0.05),提高血清IL-10含量和组织Hyp含量(P＜0.05).研究表明,APRG具有促进猪皮肤创伤愈合的作用.