Porcine Haemophilus parasuis disease, also known as Graze"s disease, is the leading bacterial infectious disease caused by H. parasuis in modern pig farms. Due to the high nutritional requirement of H. parasuis, it usually gets a false positive result through the traditional methods of plate culture. Therefore, it is necessary to establish a simple and fast detection method for this pathogen identification. This work aimed to develop a colorimetric probe using aptamer-functionalized gold nanoparticles for identifying H. parasuis. The detection mechanism is based on the color change of gold nanoparticles (AuNPs) from red to blue-purple through NaCl induction after bacteria incubation and aptamer-target binding. First, aptamer-functionalized gold nanoparticles were synthesized and characterized. Then, the best concentration of aptamers and NaCl for detection was optimized at 80 pM and 0.25 M. Under the above optimized conditions, a good linear relationship between the absorbance ratio A678/521 and H. parasuis over the range from 103 to 109 CFU/mL. The limit of detection (LOD) is 12 CFU/mL. This method is simple and rapid, results in high sensitivity and specificity, and can be used to detect actual samples.

The initial injection of PEGylated nanoparticles can activate antibodies and the complement system, leading to the accelerated blood clearance (ABC) phenomenon, characterized by reduced circulation time and abnormal liver and spleen accumulation upon re-exposure. However, PEGylation is not essential for ABC induction, as non-PEGylated nanoparticles can also trigger the similar ABC phenomenon. In this study, we found non-PEGylated nanoemulsions (CE) could accelerate the blood clearance of subsequent injection of PEGylated nanoemulsions (PE) in beagles and rats, which was independent of antibodies and the complement system, but was associated with an increase in neutrophil numbers and phagocytic activity. We propose classifying this as a “general ABC phenomenon,” broadening clinical relevance and highlighting potential immune risks of ABC phenomenon. The intensity of the ABC phenomenon correlated with the initial CE phospholipid dose in both species. Notably, larger CE particles (∼ 300 nm) induced the ABC phenomenon in beagles, while smaller particles (∼ 80 nm) with higher immunogenicity were required in rats. This suggested that beagles are more susceptible to CE-induced ABC phenomenon. The higher neutrophil proportion in beagles likely contributed to species differences in ABC phenomenon. This is the first study to report neutrophil involvement in ABC induction by non-PEGylated nanoparticles, more importantly, underscoring potential immune risks in the cross-injection of non-PEGylated and PEGylated nanoparticles during the developments and clinical applications of nano-drug delivery systems.

This study was conducted to investigate the effect of Alfalfa polysaccharide (APS) on the growth performance and immune responses of broilers. A total of 400 1-day-old Avian broiler chicks were randomly divided into four treatment groups and administered intranasal with different levels of APS (0, 2.5, 5.0 or 10.0 mg/dose) on day 1 and day 14. Growth performance, phagocytic activity, T lymphocyte transformation, and serum antibody titers against the Newcastle disease virus (NDV) vaccine were examined. During the study period of 42 days, no significant differences were observed in food intake among treatment groups (P>0.05). The APS administered groups displayed a low FCR, especially the 2.5 mg groups (P<0.05). Broilers with 2.5 mg APS displayed a much higher phagocytic activity than the control group (P<0.05). At the same time, APS enhanced the T lymphocyte transformation in response to phytohaemagglutinin (PHA). The antibody titer to NDV was not influenced by APS on day 14, while it improved on day 28 and day 42, especially in the 2.5 mg group (P<0.05). And the above data suggest that APS could improve innate and cellular immune responses in broilers.

Porcine Parvovirus (PPV) and porcine circovirus type 2 (PCV2) often have mixed infection in the process of clinical breeding, and PCV2 infection will cause immunosuppression in pigs, which is easy to stimulate or complicated with other infectious pathogens. Haemophilus parasuis (HPS) is a typical `opportunistic" pathogen, which often leads to mixed infection with PCV2 as a secondary pathogen. In order to establish a rapid and simultaneous detection of three pathogens of PPV, PCV2 and HPS, referring to the relevant genome sequence of GenBank, specific primers were designed according to the conserved region of VP2 gene of PPV, Cap gene of PCV2 and infB gene of HPS, and the amplified fragments were cloned into the vector to construct plasmid standard. Using standard samples with different dilutions as templates, adjusting primer concentration, annealing temperature and other conditions, a real-time fluorescence quantitative PCR method for PPV, PCV2 and HPS triple SYBR Green I was established. Three specific Tm peaks could be generated on the same melting curve without cross-reaction with other pathogens. The minimum detection limits of this method were 153 copies/mu L, 128 copies/mu L and 91 copies/mu L, with good specificity and repeatability, which provided technical support for rapid diagnosis of these three diseases and could be used for clinical tissue material detection.

The continuous zoonotic circulation and reassortment potential of influenza A viruses (IAV) in nature represents an enormous public health threat to humans. Beside vaccination antivirals are needed to efficiently control spreading of the disease. The previous research has shown that NOX2 involved in IAV replication, but the detailed mechanism has not been reported. In the present study we investigated the roles of NOX2 in host inflammatory response and IAV replication using a novel inhibitor GSK2795039. The drug significantly reduced H1N1 virus induced NOX2 activity and ROS release in human lung epithelial cells. The results of time course experiments suggested that GSK2795039 inhibited an early post-entry step of viral infection. Concomitantly, there was a decreased expression of pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-β and interleukin (IL)-6) in NOX2 suppressed cells. In vivo, compared with control groups, suppression of NOX2 improved the survival rate of mice infected with H1N1 virus (42.9% in GSK2795039 treated mice versus >0% of control mice) and viral burden also decreased in the GSK2795039 treated group. Thus, our data demonstrated a critical role for NOX2 in the establishment of H1N1 infection and subsequent inflammatory reactions, which suggest that GSK2795039 may be a potential therapeutic drug for IAV infection.

Actinobacillus pleuropneumoniae, Haemophilus parasuis, and Pasteurelle multocida are common pathogens of respiratory diseases in the pig industry, and they may cause secondary infections and serious economic losses to the pig industry. The clinical symptoms caused by these three pathogens are difficult to distinguish with the naked eye, and mix infections bring difficulties to the diagnosis of diseases. In this study, specific primers were designed on the basis of A. pleuropneumoniae Apx IV, H. parasuis Omp P2 and P. multocida PlpE gene. The expected amplified products ofA. pleuropneumoniae, H. parasuis, and P. multocida were 157, 120 and 305 bp, respectively. After the amplified fragment was cloned into a vector, a standard plasmid was constructed. By using the standard plasmid as template, a fluorescence quantitative PCR method for simultaneous detection ofA. pleuropneumoniae, H. parasuis, and P. multocida multiple SYBR Green I was established. Combined with melting curve analysis, the sensitivity, specificity, and repeatability were also evaluated. The results showed that the sensitivity of the method for detecting the three pathogens were 147, 145, and 61 copies/mu L. On the same melting curve that produced three specific Tm peaks, no cross reaction with other bacteria was observed, and the method demonstrated good specificity and repeatability. This method could be used for the simultaneous detection of the three pathogens, thus providing an effective detection tool for disease prevention and treatment.

Introduction:Porcine circovirus type 2 (PCV2) causes immune repression and intercurrent infections in pigs, resulting in a huge economic loss to the pig breeding industry. Additionally, the spread of PCV2 in pig farms can pollute the living environment of the residents in the farm's vicinity, which increases the rate of infections. Therefore, rapid and sensitive detection methods are needed for disease prevention and timely environmental cleaning.Methods:This research describes a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) that utilizes gold nanoparticles (AuNPs) in a functional, specific antibody labeled probe for the detection of PCV2. Due to their high specific surface area and histocompatibility, AuNPs were used as carriers of HRP labeled anti-PCV2 antibodies to amplify the detection signal.Results:Compared to conventional sandwich ELISA procedures, this method resulted in higher sensitivity (51-fold) and a shorter assay time with a limit of detection of 195 TCID50/mL. The cross-reactivity assay demonstrated that this assay was PCV2 specific.Conclusion:The amplified Ab (HRP) labeled AuNPs probe provides a sensitive analytical approach for the determination of the traces of the PCV2 antigen in early diagnosis.

This experiment was conducted to investigate the effects of weaning at 21 days of age on cecal chyme bile acids (BAs) in piglets. According to a 2 × 3 factorial design, the main factors were lactation and weaning, and the other factor was 22, 24, and 28 days of age, respectively. Piglets were randomly divided into two groups of eighteen piglets each and six piglets were selected for slaughter at 22, 24, and 28 days of age, respectively, to determine the content of different types of Bas in the intestinal lumen of the cecum. Results: (1) There was a significant interaction between weaning and age on intestinal primary Bas hyocholic acid (HCA) and chenodeoxycholic acid (CDCA) (p < 0.05), and weaning significantly increased the content of primary BAs in piglets’ intestines, which showed a trend of decreasing and then increasing with the increase in piglets’ age. (2) There was a significant interaction between weaning and age on intestinal secondary BAs deoxycholic acid (DCA), lithocholic acid (LCA), and ursodeoxycholic acid (UDCA) (p < 0.05). DCA and LCA in piglets’ intestines tended to decrease with increasing age, while UDCA showed a trend of decreasing and then increasing with increasing piglets’ age; weaning significantly increased the content of secondary BAs in piglets’ intestines. (3) There was a significant interaction between weaning and age on intestinal glycine chenodeoxycholic acid (GCDCA), taurochenodeoxycholic acid (TCDCA), and taurolithocholic acid (TLCA), but not on taurohyocholic acid (THCA), taurohyodeoxycholic acid (THDCA), and taurineursodeoxycholic acid (TUDCA) (p > 0.05). Weaning significantly increased the contents of GCDCA, TCDCA, TLCA, THDCA, and TUDCA in the intestinal tract (p < 0.05), while THCA content was not significant. In conclusion, weaning can increase the BAs content in the cecum of piglets, and there is an interaction between group and weaning age on BAs content.

Breast cancer (BC) is a very common cancer among women and one of the primary causes of death in women worldwide. Because BC has different molecular subtypes, the challenges associated with targeted therapy have increased significantly, and the identification of new therapeutic targets has become increasingly urgent. Blocking apoptosis and inhibiting cell death are important characteristics of malignant tumours, including BC. Under adverse conditions, including exposure to antitumour therapy, inhibition of cell death programmes can promote cancerous transformation and the survival of cancer cells. Therefore, inducing cell death in cancer cells is fundamentally important and provides new opportunities for potential therapeutic interventions. Lytic forms of cell death, primarily pyroptosis, necroptosis and ferroptosis, are different from apoptosis owing to their characteristic lysis, that is, the production of cellular components, to guide beneficial immune responses, and the application of lytic cell death (LCD) in the field of tumour therapy has attracted considerable interest from researchers. The latest clinical research results confirm that lytic death signalling cascades involve the BC cell immune response and resistance to therapies used in clinical practice. In this review, we discuss the current knowledge regarding the various forms of LCD, placing a special emphasis on signalling pathways and their implications in BC, which may facilitate the development of novel and optimal strategies for the clinical treatment of BC.

Actinobacillus pleuropneumoniae, Haemophilus parasuis, and Pasteurelle multocida are common pathogens of respiratory diseases in the pig industry, and they may cause secondary infections and serious economic losses to the pig industry.The clinical symptoms caused by these three pathogens are difficult to distinguish with the naked eye, and mix infections bring difficulties to the diagnosis of diseases.In this study, specific primers were designed on the basis of A. pleuropneumoniae Apx IV, H. parasuis Omp P2 and P. multocida PlpE gene.The expected amplified products of A. pleuropneumoniae, H. parasuis, and P. multocida were 157, 120 and 305 bp, respectively.After the amplified fragment was cloned into a vector, a standard plasmid was constructed.By using the standard plasmid as template, a fl uorescence quantitative PCR method for simultaneous detection of A. pleuropneumoniae, H. parasuis, and P. multocida multiple SYBR Green I was established.Combined with melting curve analysis, the sensitivity, specificity, and repeatability were also evaluated.The results showed that the sensitivity of the method for detecting the three pathogens were 147, 145, and 61 copies/μL.On the same melting curve that produced three specific Tm peaks, no cross reaction with other bacteria was observed, and the method demonstrated good specificity and repeatability.This method could be used for the simultaneous detection of the three pathogens, thus providing an eff ective detection tool for disease prevention and treatment.

In the present study, a duplex SYBR Green real-time PCR assay was developed in order to indentify Actinobacillus pleuropneumoniae and Haemophilus parasuis infection in one reaction, through a melting curve analysis. This method utilized two pairs of specific primers that allowed the amplification of highly conserved regions of A. pleuropneumoniae Apx IV and H. parasuis omp P2 gene. Reconstitution experiments were conducted by using PMD 19T plasmid in order to determine the sensitivity of the assay. The results showed that the Tm values of the melting curves of A. pleuropneumoniae and H. parasuis were 83.36±0.09oC and 76.48±0.17oC, respectively that could accurately distinguish these two pathogens. And no cross reaction were observed between other respiratory pathogens, which suggested a high specificity of two primers. The detection sensitivity of the assay was 127 and 96 copies/μL which was higher than that of the ordinary PCR detection methods. This rapid technique may present a simple, useful option for simultaneous detection of A. pleuropneumoniae and H. parasuis, which would be feasible and attractive for clinical samples diagnosis and epidemiological investigations.

为了研究副猪嗜血杆菌(Haemophilus parasuis)河南分离野毒株对昆明(KM)小鼠的致病性以及在感染KM小鼠体内组织器官的分布,将Hps血清5型标准菌株和河南分离野毒株采用灌胃和腹腔感染KM小鼠,观察2个Hps菌株对小鼠的致病性并测定Hps在感染小鼠体内组织器官的分布.结果显示,Hps河南分离野毒株和血清5型标准菌株腹腔、灌胃感染KM小鼠,在不同时间均出现发病,以腹腔感染发病症状严重、死亡率高,其中河南分离野毒株腹腔感染死亡率为2/7,血清5型标准菌株腹腔感染死亡率为1/7,死亡集中在攻毒后43～50 h;剖检死亡小鼠肉眼可见肺脏出血点、胸腔积液等病理变化.表明河南分离野毒株和标准菌株5型对KM小鼠均有致病性,为深入研究副猪嗜血杆菌的致病机制、免疫及防控该病提供科学依据.

Make sure healthy ovary or follicle is critical for extending egg laying performance in poultry. Transcription factor forkhead box L2 (FOXL2) gene have key role in regulate development of ovary. In the present research, different aged Hy-line Brown hens were maintained to explore relationships between ovarian developing and FOXL2 expression. Through histological observation, different quantities of follicles from various phases of age were observed. It was displayed that FOXL2 expression and number mature follicle were increased as the days of age increased and then decreased. In comparison, the expression of FOXL2 in hypothalamus and eyelid were remained in a relative stable level. Taken together, these data in our research establish a framework for understanding the potential functions of FOXL2 in regulate chicken ovarian developing and may provide a new perspective on the theory and practice to increase egg production or others.

高校教学中对应用技术的教育越来越重视,技能型人才在社会经济的发展中起到了越来越重要的作用,用人单位也对高校培养高素质、强能力、广适性人才的要求与呼声越来越高,这给高校人才培养提出了新的要求;适时代需要,高校教师不断探索、实践教育教学改革,在实践中启发和诱导大学生创新意识,培养其实践能力和创新能力是21世纪培养人才的必由之路,也是新形势下农业院校培养学生的重点内容.

Respiratory infections such as SARS-CoV in humans are often accompanied by mild and self-limiting hepatitis. As a respiratory disease, influenza A virus (IAV) infection can lead to hepatitis, but the mechanism remains unclear. This study aimed to investigate the occurrence of hepatitis by establishing a model for infected mice for three different subtypes of respiratory IAVs (H1N1, H5N1, and H7N2). Histological analysis was performed, and results showed increase serum aminotransferase (ALT and AST) levels and evident liver injury on days 3 and 7, especially on day 5 post infection. Immunohistochemistry (IHC) results indicated a wide distribution of IAV's positive signals in the liver of infected mice. Real-time PCR results further revealed a similar viral titer to IHC that presented a remarkedly positive correlation with histology injury. All these data showed that the mouse model suitably contributed valuable information about the mechanism underlying the occurrence of hepatitis induced by respiratory influenza virus.

A H1N1 influenza A virus infected mice model was built to evaluate the regulation mechanism of c-Jun protein in T lymphocytes. To determine the DNAzyme Dz13 efficiency in virus-infected animals, a Western-blotting assay was employed to detected total c-Jun (t-c-Jun) and phosphorylation c-Jun (p-c-Jun) levels by specific antibody. It was demonstrated that the drug-treated mice markedly suppressed the expression of t-c-Jun and p-c-Jun. Through flow cytometry and quantitative real-time polymerase chain reaction (qRT-PCR), we could find an obvious CD4+ (helper T lymphocytes) and CD8+ T (cytotoxic T lymphocyte) cell proliferation and relative cytokine (IFN-γ, IL-6 and IL-10) expression on day 3 and 6 in the virus-infected mice. The Dz13-treated mice with a c-Jun suppression displayed a down-regulation of CD4+ and CD8+ T cells in peripheral blood. At the same time, Dz13-treated group demonstrated a significant reduction of inflammatory cytokine (IFN-γ, IL-6 and IL-10) expression. In conclusion, c-Jun protein regulates CD4+ and CD8+ T cell proliferation and inflammatory cytokine expression in H1N1 influenza A virus infected mice.

Porcine epidemic diarrhea virus (PEDV) is a devastating swine infectious disease. Development of high-performance methods to diagnose and evaluate viral immune status remains very important to control PEDV. Here, a recombinant S1 protein-based indirect enzyme-linked immunosorbent assay (rpS1-iELISA) was developed to monitor IgA antibody in the colostrum. Optimized cut-off value of the rpS1-iELISA was determined as 0.448. Results yielded a sensitivity of 96.87% and a specificity of 100.0%. Repeatability tests indicated that the coefficients of variation of the colostrum samples within and between runs were both less than 8%.Test results of 523 field colostrum samples showed that the rpS1-IELISA had excellent agreement with immunofluorescence assay (kappa = 0.958) and better test performance than a commercial ELISA kit. This test will aid in future diagnostics and assessment of the protective levels of mucosal immune response against PEDV by measuring IgA levels in the colostrum.

M细胞又称微皱褶细胞(microfold cell)或膜性细胞(membranous cell),是组成黏膜免疫屏障的一类重要上皮细胞,在黏膜免疫和病原入侵方面发挥重要功能.论文就M细胞的形态、功能及其在病原入侵中的作用进行综述,以期拓展对M细胞的认识以及引起兽医领域对M细胞研究的关注.