Objective: To investigate the role of osteoclast-derived apoptotic bodies (OC-ABs) in osteoarthritis (OA), specifically their impact on subchondral bone remodeling and disease progression, and to explore potential therapeutic strategies targeting OC-AB-induced pathways. Methods: We utilized a mouse model of anterior cruciate ligament transection (ACLT) to simulate post-traumatic osteoarthritis (PTOA). Levels of OC-ABs were assessed in subchondral bone and correlated with OA severity. Additionally, apoptotic body-deficient MRL/lpr mice were analyzed to evaluate the direct contribution of OC-ABs to OA progression and subchondral bone remodeling. The involvement of OC-ABs in osteogenesis was further examined using mesenchymal stem cells (MSCs), with a focus on the RANKL reverse signaling pathway. The therapeutic potential of rapamycin to counteract OC-AB effects was tested. Results: Increased OC-AB accumulation in subchondral bone was positively correlated with OA severity in ACLT-induced mice. Apoptotic body-deficient MRL/lpr mice demonstrated slower OA progression and maintained more stable subchondral bone architecture, indicating a pathogenic role of OC-ABs in OA. OC-ABs significantly stimulated osteogenesis in MSCs via the RANKL reverse signaling pathway. Treatment with rapamycin effectively reversed OC-AB-induced subchondral bone formation, mitigated OA progression, and inhibited the RANKL reverse signaling pathway. Conclusion: OC-ABs play a critical role in exacerbating OA by promoting subchondral bone remodeling via the RANKL reverse signaling pathway. Rapamycin presents as a promising therapeutic agent capable of mitigating OC-AB-driven pathology, highlighting new avenues for targeted OA treatment.

The rapid growth of animal husbandry has led to significant environmental challenges, making the management of livestock and equine waste a pressing social issue. Based on the utilisation of livestock waste, the present article establishes a "four-in-one" ecological animal husbandry planting-and-raising mode in combination with agro-ecosystems. Using a recycling agricultural park as a case study, the ecological and economic benefits of the combined model were validated through analyses of energy flow and value changes. Under the combined planting-and-raising mode, the light energy utilisation rate reaches 1.02%. The energy value output of the recycled agricultural park was 1.83 times higher than that of the monoculture mode. By utilising livestock and equine waste for biogas production, the average annual manure processing volume reaches 9212.44 tonnes, yielding an average of 25.8 & times; 10<^>4 cubic meters of biogas annually. Replacing part of the chemical fertiliser with manure for planting silage maize increased the harvest yield under small-field management to 3847kg/665 m(2), which was 147kg/665 m(2) higher than in the field without manure application. Conclusively, effective utilisation of livestock wastes should follow a combined planting-and-raising model to promote agro-ecosystem recycling, providing a reliable solution to reduce environmental pollution and enhance ecological benefits.

Ulcerative colitis (UC) can cause severe oxidative stress in the colon, which can lead to tissue damage and an imbalance in the normal gut microbiota. Ellagic acid (EA) is one of the main types of plant polyphenols with improved pharmacological effects such as antioxidant, anti-inflammatory, and antibacterial properties. However, currently, the studies on the impact of EA on the gut microbiota and its potential to alleviate UC in mice through the ROS/NLRP3 pathway are limited. In this study, dextran sodium sulfate (DSS) was used to construct a UC mouse model, which was then treated with EA as an intervention for UC. The results revealed that EA alleviated the trend of liver, spleen, and weight changes in UC mice and improved colon oxidative stress, inflammation, and pathological damage. Mechanistically, DSS-induced UC indicated a significant increase in ROS/NLRP3 pathway-related factors, whereas EA intervention activated the Nrf2 pathway to reduce these factors. Furthermore, the DSS group had a reduced abundance of Firmicutes (59.02%) and an increased abundance of Bacteroides and Proteobacterium by 1.8 times and 10.16%; however, EA intervention reversed these changes, thus alleviating UC. The findings of this study revealed that EA could significantly enhance the composition of gut microbiota in UC and reduce the inflammatory response, colonic damage as well as oxidative stress caused by DSS by regulating the ROS/NLRP3 pathway. These results provide novel perspectives on the prevention and treatment strategies of UC and highlight the therapeutic benefits of EA in managing colitis.

Duck enteritis virus (DEV) may lead to vascular injury, gastrointestinal mucosal erosion, lymphoid organ injury, and Polyinosinic-polycytidylic acid (Poly I:C) has an antiviral effect by inducing low levels of interferon. The purpose of this study was to explore the pathogenesis of DEV-induced intestinal injury in ducks and to verify the therapeutic effects of different concentrations of Poly I:C. In this study, duck enteritis model was established by infecting healthy Pekin ducks with DEV. Duck intestinal tissues were extracted from normal control group, model group, and treatment group with different doses of Poly I:C. In vivo, HE and TUNEL staining were used to observe the morphological changes and apoptosis. In vitro, the proliferation and apoptosis of duck intestinal epithelial cells were evaluated by MTT assay, TUNEL staining, and flow cytometry. The results showed that Poly I:C protected ducks from DEV toxicity by improving intestinal morphology and inhibiting apoptosis. In addition, the antiviral effect of Poly I:C on DEV was found in a dose-dependent manner, with a more relatively obvious effect at a high dose of Poly I:C. All in all, these results demonstrated that Poly I:C played a vital role in the apoptosis induced by DEV in ducks and modest dose of Poly I:C treatment worked well and may provide important reference for the development of new antiviral drugs in the future.

将100只健康30日龄雏鸭随机分为试验组(Ⅰ组,50只)和对照组(Ⅱ组,50只),进行鸭肠炎病毒(duck enteri-tis viral,DEV)检测.试验组每只雏鸭腿部肌肉注射0.2 mL(1 000 LD50),Ⅱ组注射无菌生理盐水0.2 mL,分别于感染后12,24,36,48,72,84,96 h无菌采集各组织样本(肌肉、胸腺、十二指肠、法氏囊、心脏、肝脏、脾脏、肺脏、肾脏、脑),利用荧光定量PCR和ELISA检测方法确定Ⅱ组未发生感染、Ⅰ组成功感染后,所有样品-80℃保存备用.同时,对各时段各组织中JAK2、STAT3和SOCS1基因的转录水平与蛋白质表达量进行检测,并对十二指肠进行转录水平和蛋白表达水平验证.结果显示,与Ⅱ组相比,JAK2、STAT3和SOCS1基因在DEV感染鸭机体组织中转录水平差异显著(P＜0.05).JAK2和STAT3蛋白表达水平低于Ⅱ组,差异显著(P＜0.05),而SOCS1蛋白表达量水平高于Ⅱ组,差异显著(P＜0.05);取十二指肠组织进行转录组测序和Western blot验证,结果与荧光定量PCR和ELISA检测结果一致.结果表明,感染DEV对雏鸭各组织JAK-STAT信号通路相关基因JAK2、STAT3和SOCS1基因的转录水平均有不同程度的影响.感染DEV对鸭机体组织JAK-STAT信号通路SOCS1基因的影响是在蛋白水平的上调,而非转录水平,同时SOCS1基因对JAK2、STAT3基因的抑制作用也是在蛋白水平.

为建立鸭NF-kB1(nuclear factor-kappa B,NF-kB)基因的荧光定量PCR检测方法,并以建立的方法检测鸭胚成纤维细胞(duck embryo fibroblasts,DEF)在感染鸭肠炎病毒(DEV)后NF-kB1基因随时间变化转录表达量的变化情况.根据GenBank上NF-kB1基因保守序列设计特异性引物,以鸭胚成纤维细胞mRNA提取样本反转录为cDNA,进行NF-kB1基因克隆质粒构建,以此为模板建立鸭NF-kB1基因荧光定量PCR检测方法并进行特异性、重复性和敏感性试验,并利用建立的方法DEF感染DEV后NF-kB1基因随时间变化的转录变化进行检测.结果显示:建立的鸭NF-kB1基因荧光定量PCR方法标准曲线呈现典型的S型,方程为y=-3.12x+44.086(R2=1),扩增效率为109.2％;其熔解温度Tm值为(83.5 ±0)℃,曲线呈特异性单峰,批内变异系数小于0.3％,批间变异系数小于0.2％;检测灵敏度可达到1.79个拷贝.DEF细胞在感染DEV后NF-kB1基因随时间改变转录水平变化无规律,但整体表达水平高于正常细胞,差异显著(P ＜0.05),36～84 h NF-kB1基因的转录水平与正常细胞中差异极显著(P ＜0.01).本研究成功建立了鸭NF-kB1基因荧光定量PCR检测方法,并对DEF细胞感染DEV后NF-kB1基因的转录水平进行了研究,为后续实验研究提供了技术和数据支撑.

为探究NF-κB信号通路对鸭肠炎病毒(DEV)增殖的影响,本实验分别以不同终浓度的NF-κB信号通路激活剂LPS(2.0μg/mL、4.0μg/mL、6.0μg/mL)和抑制剂SN50(25.0μg/mL、50.0μg/mL、75.0μg/mL)处理鸭胚成纤维(DEF)细胞后,采用CCK8法分析LPS或SN50对DEF细胞活性影响;利用不同浓度的LPS或SN50预处理DEF细胞4 h后,再于12 h～120 h后收获细胞和上清液(未感染DEV);上述经LPS或SN50预处理细胞4 h后接种DEV(MOI 0.01),12 h～120 h后(每间隔12 h)分别收获细胞及上清液,采用荧光定量PCR方法分别检测上述未感染和感染DEV的DEF细胞中NF-κB1基因和DEV NP基因的转录水平;采用ELISA方法分别检测上述未感染和感染DEV的DEF细胞上清液中NF-κB信号通路关键因子(IL-1β、IL-6和MyD88)的表达水平.结果显示:不同浓度的LPS和SN50处理对DEF细胞均无明显毒性作用.荧光定量PCR结果显示,经不同浓度LPS预处理,均能有效提高DEF细胞中NF-κB1基因的转录水平,其中4.0μg/mL LPS处理的DEF细胞中NF-κB1基因转录水平最高;而在LPS预处理再感染DEV后,4.0μg/mL LPS和6.0μg/mL LPS处理的DEF细胞中NF-κB1基因转录水平整体上调(p<0.05),2.0μg/mL LPS处理的DEF细胞中NF-κB1基因的转录水平整体下调(p<0.05);不同浓度的SN50预处理后,未感染和感染DEV的DEF细胞中,均以50.0μg/mL SN50处理的细胞中NF-κB1基因的转录水平下调效果最好(p<0.05).LPS预处理DEF细胞后感染DEV,12h～84 h DEV NP基因的转录水平均受到明显抑制(p<0.01),84 h后2.0μg/mL和6.0μg/mL LPS均会促进DEV NP基因的转录水平;而经SN50预处理DEF细胞后感染DEV,细胞中DEV NP基因的转录水平均显著下降(p<0.01).ELISA结果显示,经LPS预处理后,不感染或感染DEV,DEF细胞中IL-6表达量整体稍呈下降趋势(p>0.05),而IL-1β和MyD88的表达则呈无规律变化;经SN50预处理后不感染或感染DEV,DEF细胞中IL-1β、IL-6和MyD88的表达均无规律变化.以上研究结果表明,不同浓度LPS均可促进正常DEF细胞中NF-κB1基因的转录,而DEV则可以阻断低浓度LPS(2.0μg/mL)的这种促进作用.不感染或感染DEV,SN50均于高浓度(50μg/mL和75μg/mL)时才能有效降低NF-κB1基因的转录水平;不同浓度的LPS或SN50均对NF-κB通路关键因子的表达基本无影响;LPS在DEV感染后期才能促进其增殖,而SN50则可以有效抑制DEV的增殖.本研究为阐明NF-κB信号通路与DEV之间的相互作用关系奠定了实验基础..

文章旨在分析Cofilin在鸭肠炎病毒感染过程中的影响.以受精鸭蛋、鸭肠炎病毒GZ株为样本,运用相关试剂及仪器,开展试验分析.通过对DEF细胞进行转染处理,共计4个干扰质粒转染成功,且顺利表达,沉默率最高为86型.结论:DEV感染小组及正常小组各个组织Cofilin2基因于相同时间点转录变化一致性强,筛选获取沉默效应显著的Cofifilin-shRNA-86,其能达到促进DEV增殖效果.

为了解引起鲤鱼皮肤溃疡的病原分子分型特征及其毒力基因,从患溃疡症鲤鱼体内分离到1株致病性菌株GZGY2 019,鉴定其为气单胞菌,并运用多位点序列分型(MLST)方法进行分类研究,利用PCR和测序方法分析6种毒力基因Aer、Ser、Alt、Lip、Act和Hly.通过对分离株的16S rDNA基因进行分析,确认其属于杀鲑气单胞菌.分离株对复方新诺明等9种抗菌药物耐药,毒力基因扩增结果显示:分离菌能检出气溶素基因(Aer)等6种毒力基因.基于气单胞菌6个管家基因分析,与MLST数据库中等位基因序列比对,发现其属于新的序列型(ST),隶属于杀鲑气单胞菌无色亚种.这一研究对发病鲤鱼的病因进行了准确诊断,为该病的防治与分子流行病学研究提供了依据.

Duck viral enteritis (DVE) is a lethal viral disease caused by duck enteritis virus (DEV) via an unknown mechanism. This study explores the relationship between Chinese standard challenge strain DEV (DEV-CSC)-induced apoptosis and endoplasmic reticulum stress (ERS) in duck embryo fibroblast (DEF) cells. Here we examined changes in Ca2+ concentration, cell proliferation, apoptosis, and the differential expression of C/EBP homologous protein (CHOP), glucose regulatory protein 78 (GRP78), and activating transcription factor 6 (ATF6) in infected cells. The results revealed that DEV-CSC infection significantly decreased Ca2+ concentration, suppressed cell viability, and induced apoptosis in DEF cells. Further experiments also demonstrated that DEV-CSC infection significantly upregulates CHOP, GRP78, and ATF6 expression. In addition, we show that the addition of ethylenediaminetetraacetic acid (EDTA) reverses the induction of apoptosis and the ERS mediated inhibition of cell viability in DEF cells associated with DEV-CSC infection. Therefore, we can conclude that infection with DEV-CSC induces apoptosis and ERS reducing the viability of DEF cells via the regulation of Ca2+. These findings may provide a new target for the treatment of DVE.

为了建立鸭白细胞介素-1β(IL-1β)基因的实时荧光定量PCR检测方法,并检测鸭机体器官/组织中IL-1β基因转录水平,试验根据GenBank上鸭IL-1β基因序列设计特异性引物,以鸭IL-1β基因克隆质粒为模板,SYBR Green I为荧光染料,建立鸭IL-1β基因实时荧光定量PCR检测方法,并用该方法检测鸭机体器官/组织中IL-1β基因的转录水平.结果 表明:建立的鸭IL-1β基因实时荧光定量PCR方法的熔解曲线为特异性单峰,标准曲线方程为y=-3.329x+39.731,扩增效率为99.7％,相关系数为1,批内重复变异系数小于1.00％,批间重复变异系数小于2.00％;实时荧光定量PCR方法的检测敏感性是普通PCR方法检测敏感性的1 000倍;以该方法进行检测发现,鸭的法氏囊、肺脏、肝脏、肌肉、脑、脾脏、肾脏、十二指肠、心脏和胸腺中IL-1β基因mRNA含量分别为428.45,973.43,1 394.19,568.22,551.54,839.91,2 586.22,1 161.92,459.17,3 276.16 copies/μL,其中胸腺中含量最高,而法氏囊中含量最低.说明试验建立的鸭IL-1β基因实时荧光定量PCR检测方法具有重复性好和敏感性高等特点,可用于鸭机体器官/组织中IL-1β基因转录水平的检测分析.