动物传染病已经严重威胁到动物和人类的生存与发展,并逐渐成为全球瞩目的共性问题.对动物传染病的研究,已成为当前生命科学研究中最重要的一部分.国内动物传染病研究工作近年来得到了长足发展,取得了一系列重要成果.本文以近三届(2015-2019年)中国畜牧兽医学会动物传染病学分会全国学术研讨会的论文与报告为基础,归纳数据、分析规律,剖析"十三五"期间国内动物传染病研究的特点和规律,为动物传染病学科的建设和发展提供参考.动物传染病的研究范围、疫病种类更广泛,对新发传染病更加关注;机制研究更深入;疫病研究具有明显的地区特点,研究机构区域化特征明显.

Acrylamide (AA) constitutes an important industrial chemical agent and well-known neurotoxin. However, the mechanism underlying AA-mediated neurotoxicity is extremely complicated and controversial. In this study, we found that activation of the NLR family pyrin domain containing 3 (NLRP3) inflammasome and its subsequent downstream inflammatory responses plays an important role in AA-induced neurotoxicity mechanisms. In vitro experiments revealed that AA (2.5 mM) induced BV2 microglial cytotoxicity and triggered NLRP3 inflammasome activation along with downstream proinflammatory cytokine interleukin-1β and interleukin-18 expression. Treatment with inhibitor or NLRP3 siRNA efficiently protected BV2 microglial cells against AA-induced cytotoxicity and reversed NLRP3 inflammasome activation and its mediated inflammatory reaction. Similarly, AA exposure (50 mg/kg) for 10 consecutive days caused significant activation of NLRP3 inflammasomes and neuroinflammation in C57BL/6 mice, whereas inhibiting these effects through specific NLRP3 inflammasome blocker MCC950 (5 mg/kg) intervention or NLRP3 knock-out significantly ameliorated AA-induced ataxia, cerebellar Purkinje cells degeneration, and apoptosis. Furthermore, we demonstrated that antagonism of NLRP3 could also up-regulate the Nrf2 signalling pathway and related antioxidant genes. In conclusion, our findings indicate that activation of the NLRP3 inflammasome pathway is involved in AA-induced neurotoxicity, whereas MCC950 treatment or NLRP3 knock-out could effectively protect against AA-induced neurotoxic injury through the inhibition of neuroinflammation and activation of the Nrf2 antioxidant pathway. Therefore, the NLRP3 inflammasome might serve as a promising therapeutic target, with drugs designed to specifically inhibit this pathway potentially providing new avenues for preventing or ameliorating AA poisoning.

FTY720, a S1P-receptor modulator, has shown to be effective in several transplant and autoimmune disease models, via modulating lymphocyte homing into secondary lymphoid organs (SLOs), and thereby reducing these cells in peripheral blood. ASP0028, a newly developed S1P1/S1P5-selective agonist, presented comparable efficacy to FTY720 and wider safety margins than FTY720. In this study, we assessed the efficacy and safety of ASP0028 co-administered with suboptimal-dose of tacrolimus in the Cynomolgus monkey renal transplantation model. Seven animals in group-1 or group-2 received mono-tacrolimus 1.0 mg/kg once a day (QD), or ASP0028 0.6 mg/kg plus tacrolimus 1.0 mg/kg QD, respectively. Eight animals in group-3 received ASP0028 1.2 mg/kg plus tacrolimus 1.0 mg/kg QD. The allograft median survival time (MST) in group-2 and group-3 were significantly extended to 41 and 61.5 days, versus that of 28 days in group-1 (p = 0.036 and 0.001, respectively). ASP0028 administration remarkably reduced absolute numbers of peripheral lymphocytes, particularly subsets of CD4+/ or CD8+/naive and central memory cells, CD4+/Treg cells, and to a lesser extent on B cells, but not CD4+/ or CD8+/effector memory cells and NK cells. These data show ASP0028 combined with suboptimal-dose of tacrolimus effectively prolongs renal allograft survival in nonhuman primates (NHPs) with well tolerated safety, supporting its further investigation to optimize CNI-sparing regimens.

Objective This study aims to investigate the effects of high-fat diet rich in perilla oil on the expression of key genes that regulate hepatic VLDL synthesis in obese rats .Methods Sixty healthy male 5-week old SD rats were randomly divided into 2 groups.The rats in the normal control group (NC, n=12) were given normal diet, and the rats in the high fat group ( HF, n=48) were given a pure high fat diet in order to induce rat models of obesity .In the intervention period, the obesity model rats were randomly divided into 4 subgroups including consistent high fat group (CHF) and three intervention groups depending on perilla oil substitution rate of lard in CHF:20%PO, 50%PO and 100%PO.The serum triglyceride (TG) of the rats was measured after 4 weeks.Real-time PCR was applied to measure microsomal triglyceride transfer protein ( Mtp) and apolipoprotein B ( Apob) mRNA, and western blot assay was used for detecting the expression of MTP and APOB in the liver .Results Compared with the NC group , the CHF rats exhibited significantly high fat deposi-tion.The serum TG was markedly higher and the MTP and APOB were decreased at gene and protein levels in the CHF group compared with the NC group .After the intervention , PO remarkably reduced the level of serum TG and decreased he-patic fat deposition as it showed by pathological examination .At the gene and protein levels , MTP and APOB were upregu-lated by PO to different degrees .Conclusions All the three PO intervention can promote VLDL synthesis and secretion , and decrease the hepatic fat deposition in the obese rats .Furthermore , PO upregulates the expression of MTP at gene and protein levels in a dose-dependent manner .

Background. Blocking the CD40-CD154 signal pathway has previously shown promise as a strategy to prevent allograft rejection. In this study, the efficacy of a novel fully human anti-CD40 monoclonal antibody-ASKP1240, administered as a monotherapy or combination therapy (subtherapeutic dose of tacrolimus or mycophenolate mofetil), on the prevention of renal allograft rejection was evaluated in Cynomolgus monkeys.Methods. Heterotopic kidney transplants were performed in ABO-compatible, stimulation index 2.5 or higher in the two-way mixed lymphocyte reaction monkey pairs. Animals were divided into 12 groups and observed for a maximum of 180 days. Histopathologic, hematology, and biochemistry analyses were conducted in all groups. Cytokine release (interleukin [IL]-2, IL-4, IL-5, IL-6, tumor necrosis factor, and interferon-gamma) was investigated in several groups.Results. ASKP1240 prolonged renal allograft survival in a dose-dependent manner in monotherapy. Low-dose (2 mg/kg) or high-dose (5 mg/kg) ASKP1240, in combination with mycophenolate mofetil (15 mg/kg) or tacrolimus (1 mg/kg), showed a significantly longer allograft survival time compared with monotherapy groups. No obvious side effects including drug-related thromboembolic complications were found. Cytokine release was not induced by ASKP1240 administration.Conclusion. The present study indicates that ASKP1240, alone or in combination with other immunosuppressive drugs, could be a promising antirejection agent in organ transplantation.

iRhom1 and iRhom2 are inactive homologues of rhomboid intramembrane serine proteases lacking essential catalytic residues, which are necessary for the maturation of TNFα-converting enzyme (TACE). In addition, iRhoms regulate epidermal growth factor family secretion. The functional significance of iRhom2 during mammalian development is largely unclear. We have identified a spontaneous single gene deletion mutation of iRhom2 in Uncv mice. The iRhom2Uncv/Uncv mice exhibit hairless phenotype in a BALB/c genetic background. In this study, we observed dysplasia hair follicles in iRhom2Uncv/Uncv mice from postnatal day 3. Further examination found decreased hair matrix proliferation and aberrant hair shaft and inner root sheath differentiation in iRhom2Uncv/Uncv mutant hair follicles. iRhom2 is required for the maturation of TACE. Our data demonstrate that iRhom2Uncv cannot induce the maturation of TACE in vitro and the level of mature TACE is also significantly reduced in the skin of iRhom2Uncv/Uncv mice. The activation of Notch1, a substrate of TACE, is disturbed, associated with dramatically down-regulation of Lef1 in iRhom2Uncv/Uncv hair follicle matrix. This study identifies iRhom2 as a novel regulator of hair shaft and inner root sheath differentiation.

目的 比较黑线仓鼠及其白化突变系背部皮肤蛋白表达的差异,寻找差异蛋白质,从蛋白质水平探讨白化病的发生机制.方法 应用双向凝胶电泳技术分离出差异蛋白质,用质谱法分析其结构与组成,通过蛋白质数据库确定差异蛋白的功能.结果 从64个表达差异蛋白斑点中发现33个显著差异的蛋白点,其中又有14个差异点匹配到了有意义的蛋白质.14个差异点共鉴定出11个差异蛋白质,这些差异蛋白质按功能可分为4类:(1)糖代谢相关蛋白;(2)运输蛋白;(3)细胞骨架蛋白;(4)其他蛋白.结论 黑线仓鼠与其白化突变系背部皮肤蛋白表达存在明显差异,其中一些蛋白与白化病发生相关,并可能成为白化病致病机理研究的分子标志物和药物治疗靶向位点.

Objective To obtain a stably inherited Sprague-Dawley rat model of congenital umbilical hernia by in-breeding, and to observe the structure of umbilical hernia and treat it surgically.Methods Congenital umbilical hernia rats were fostered by full-sib mating.The birth number and umbilical hernia quantity were recorded, and the umbilical hernia rate of rats was analyzed.Six female and 6 male rats with congenital umbilical hernia of 6-month aged F2 generation were selected randomly, among which 2 female and 2 male rats were examined anatomically, and the rest rats underwent surgical suture. Results The umbilical hernia rate was increased along with the increasing inbreeding coefficient, and the rats of F12 and F13 generations were all with congenital umbilical hernia.The umbilical hernia rate in female rats was significantly higher than that in male rats based on the total number of rats from F1 to F13 generation (c2 =11.1, P=0.001).Female and male rats had the same structure of umbilical hernia, and all rats recovered 3-4 weeks after surgery without recurrence.Conclusion After 13 consecutive generations of full-sib mating, a rat model of congenital umbilical hernia with stable genetic properties is successfully established.

Background The purpose of this study was to evaluate the serum concentration of ASKP1240 (pharmacokinetics [PK]) and the CD40 occupancy of ASKP1240 (pharmacodynamics [PD]) in normal and renal transplanted Cynomolgus monkeys to clarify the PK/PD relationship. Methods In a 70-day study, two ASKP1240 doses (2 and 5 mg/kg) were evaluated in normal and transplanted monkeys. Full doses were administered during the induction phase, and half doses were administered during the maintenance phase. The PK and PD were assessed using ELISA and FACS assays. Results The serum concentration and receptor occupancy of ASKP1240 reached their maximum levels rapidly after the first dose and remained at an almost saturated rate during the induction phase. They then decreased gradually during the maintenance phase in all of the groups. The serum concentration and duration of full receptor occupancy were dose dependent in the normal and transplanted monkeys. On day 70 after therapy with 5 mg/kg ASKP1240, the transplanted monkeys presented a significantly lower occupancy of the CD40 receptors compared with the normal animals (5.5%±14.1% vs. 72.8%±3.4%). The serum concentration of ASKP1240 was also strongly correlated with the occupancy of the ASKP1240 receptors. Conclusion This study showed strong positive PK/PD relationships in renal transplanted and normal monkeys. The results may thus serve as a guide for optimal dosage and timing of ASKP1240 therapy in clinical trials and will propel the translation of ASKP1240 therapeutics from the bench to preclinical and clinical trials.

Objective To study the effects of mouse UNCV protein on apoptosis in HeLa cells.Methods The recombinant plasmids containing Uncv gene of BALB/C mice were transfected into HeLa cells.The HeLa cells stably overexpressing UNCV protein were selected.The effects of overexpressed UNCV protein on apoptosis in HeLa cells induced by doxorubicin and serum starvation were measured by cell counting and flow cytometry.Results HeLa cells stably overexpressing UNCV protein were obtained.The results of cell counting and flow cytometry showed that the overexpression of UNCV protein inhibited apoptosis in HeLa cells induced by serum starvation.The effects of UNCV protein on apoptosis in HeLa cells induced by doxorubicin were not significantly different.Conclusions Overexpression of UNCV protein inhibits apoptosis in HeLa cells induced by serum starvation.

Objective Using loop-mediated isothermal amplification(LAMP) assay to develop a rapid,convenient,sensitivity and specificity of enterotoxigenic Escherichia coli(ETEC) detection method,and evaluate the specificity and sensitivity of this method,for detection and diagnosis of bacterial diarrhea in laboratory animals.Methods According to the published E.coli LT toxin gene sequences(S60731.1) to design PCR and LAMP primers,and to compare the specificity and sensitivity of LAMP with that of PCR.Result The detection limit of the established LAMP method were 100 pg,its sensitivity was 10 times higher than that of PCR,and had a high specificity.Using PCR and LAMP to detect 27 feces samples of monkey diarrhea,the results of LAMP(within 60 min) were the same as that of PCR,with a positive rate of 33.3%.But the positive rate of LAMP(within 90 min) was 92.6%,about 3 times higher than that of PCR.Conclusions We have established an ETEC LAMP detection method,which is highly specific,sensitive,convenient,suitable for rapid detection of ETEC clinical samples.

We detected Bartonella quintana in 48.6% of captive rhesus macaques from an animal facility in Beijing, China. Prevalence of infection increased over the period of observation. Our findings suggest that macaques may serve as reservoir hosts for B. quintana and that Pedicinus obtusus lice might act as efficient vectors.

Enterovirus 71 (EV71) is the major causative agent of hand, foot, and mouth disease and induces fatal neurological complications. In recent years, this virus has become a major threat to public health in the Asia–Pacific region, while no effective antiviral therapies and vaccines are currently available. In this study, we constructed and characterized for the first time an infectious full-length EV71 cDNA clone derived from the SHZH98 strain, which was the first subgenotype C4 strain isolated in China. Our data demonstrate that the rescued EV71 viruses exhibited growth kinetics in vitro and morphologies similar to those of the BrCr-TR strain and reached a maximum titer of 107.5 TCID50/ml. Although the rescued viruses were able to infect suckling mice, no typical symptoms of EV71 infection were observed for up to 18 days post-inoculation. Taken together our research provides an important tool to study the epidemic strains of EV71 in the Asia–Pacific region and promote the development of vaccines.

The detection of humoral immune responses is the method of choice for the rapid identification of B virus-infected animals.To establish the foundation for rapid diagnosis tech and to prepare for vaccine develop,extracellular domain of monkey B virus glycoprotein D was expressed and its diagnostic potential was evaluated.In this study,the extracellular domain gene of glycoprotein D was amplified by PCR and cloned into the prokaryotic expressive vector pET-28b(+) after sequencing.Recombinant plasmid transfected into E.coli BL21(DE3) and protein expression was induced by IPTG.The purified protein was identified by His antibody and B virus positive serum,and its diagnostic value was evaluated by ELISA.Results showed that the recombinant protein had a Mr of 48 kD,accounted for 35% total proteins,and expressed in form of inclusion body.Western blot showed that the recombinant protein could react specifically to the monkey B virus positive sera and anti-histidine monoclonal antibody.The indirect ELISA of 32 standard positive sera and 30 negative sera showed the detection rate were 93.75% and 100% respectively.These data indicate that recombinant glycoprotein D from prokaryotic expression system had perfect reactionogenicity,which would can be used as the candidate antigen of monkey B virus serological testing.

Objective:To observe the protective effects of Baohuoside aglycone(BA) in renal ischemia/reperfusion(I/R) injury in the rat.Methods:A total of 39 healthy male adult Sprague-Dawley(SD) rats were evaluated in three groups.Group Ⅰ(sham),Group Ⅱ(renal I/R injury treated with vehicle),Group Ⅲ(renal I/R injury treated with BA).The model of renal ischemia/reperfusion injury was induced with left renal pedicle occlusion for 50 min followed with reperfusion and then right nephrectomy was performed under anesthesia.The level of BUN,SCr,CrCI were determined in every rat on day 3,7,and 14 post-operatively.Three rats in every group were sacrificed on day 3 after operation,whose left kidney was harvested for histopathological examinations.Results:After three days of operation,the serum concentrations of SCr and BUN in Group Ⅲ were significant lower in comparison with that in Group Ⅱ.Seven days after operation,the SCr and BUN in BA treat group had resumed as same level as that in Group Ⅰ(sham group).On the contrary,renal function in Group Ⅱ recovered until 14 days postoperatively.Conclusion:Baohuoside aglycone could reduce the I/R injury in rat model.It can be a favorable choice for protection of renal I/R injury in clinic.

In the mouse embryo, the aorta-gonad-mesonephros (AGM) region is considered to be the sole location for intraembryonic emergence of hematopoietic stem cells (HSCs). Here we report that, in parallel to the AGM region, the E10.5-E11.5 mouse head harbors bona fide HSCs, as defined by long-term, high-level, multilineage reconstitution and self-renewal capacity in adult recipients, before HSCs enter the circulation. The presence of hemogenesis in the midgestation head is indicated by the appearance of intravascular cluster cells and the blood-forming capacity of a sorted endothelial cell population. In addition, lineage tracing via an inducible VE-cadherin-Cre transgene demonstrates the hemogenic capacity of head endothelium. Most importantly, a spatially restricted lineage labeling system reveals the physiological contribution of cerebrovascular endothelium to postnatal HSCs and multilineage hematopoiesis. We conclude that the mouse embryonic head is a previously unappreciated site for HSC emergence within the developing embryo.

Objective To detect the function of I BHV-1 TK-/gE-/ EGFP+ gene-deleted mutant virus strain,find a new method to prevent infectious bovine rhinotracheitis(IBR),the charaction of TK-/gE-/ EGFP+ gene-deleted mutant virus strain was further studied.Methods Southern hybridization,western dot blot and Plaques test were employed in this investment.Results TK gene is deleted and the mutant cannot produce glycoprotein E when gE gene is deleted.The diameter of mutant strain much smaller than that of field strain,the mutant strain grows on MDBK cells and the virus titer can keep 106.0TCID50/0.1 mL for more than 40 hours,which indicated that the mutant strain inherit instantly.Conclusion The gene-deleted mutant virus strain was successful constructed in this study,and the mutant virus strain was stable.This will be useful for the further study of IBR.

Objective To observe the protective effects of Baohuoside aglycone(BA) in myocardial Infarction of ischemia/reperfusion(I/R) injury in the rat.Methods Myocardial ischemia/reperfusion injury model was induced by ligation the left descending anterior coronary artery(LAD).Thirty-six SD rats were divided into three groups: sham group,I/R injury group and BA treated group,each contained 12 rats respectively.The serum superoxide dismutase(SOD) was tested 3 days after surgery and the rate of the infraction muscle weight was measured 21 days after surgery.Result The serum SOD in BA treated group is higher than that in I/R injury group(6.54±0.41 vs.5.64±0.68 P=0.003).The rate of the BA treated group is lower than that in I/R injury group(9.03± 0.22% vs.31.83±2.09%,P<0.01).Conclusion Due to the reduction of the injury of free radicals,Baohuoside aglycone possesses cardioprotective effect in prevention of ischemia/reperfusion injury and reduce the myocardial infraction.

Objective To investigate the effect of paclitaxel on the Cynomolgus monkey and human liver microsomes CYP1A2,CYP2A6 and CYP3A4 enzyme activity.Methods To take Phenacetin,testosterone and coumarin medicine as a tool,we adopted the Cynomolgus monkey and human liver microsomes to establish the metabolic system in vitro.The different concentrations of paclitaxel were incubated with the three kinds of substrate in liver microsomes system respectively.To use HPLC to measure the metabolites output included paracetamol,6β-hydroy testosterone and 7-hydroxycoumarin,the IC50 value was calculated to evaluate the effect of the metabolism of CYP1A2,CYP2A6 and CYP3A4.Results The IC50 values about Cynomolgus monkey liver microsomes were 570 ± 5.9 μmol/L,140 ± 2.9 μmol/L and no effect.The IC50 values about human liver microsomes were 193 ± 6.6 μmol /L,253 ± 3.6 μmol /L and 24 ± 1.6 μmol /L.Conclusions Paclitaxel inhibits the activity of CYP1A2 and CYP3A4 liver microsomes,it has little impact on theCYP2A6 enzyme.Paclitaxel has weak inhibitory on human liver CYP1A2 and CYP3A4 enzyme activity,but it has strong inhibition on the CYP2A6 enzyme activity,the results give some mirror to the safety of clinical medication.

Objective To develop a proper animal models representing human diabetes,the efficiency and doses of alloxan in male and female animals was investigated.Methods Alloxan were administered to beagle dogs and mice with different dosage.Blood glucose was tested after treatment at the 3rd,7th,14th,and 21th day,respectively.Results The different effects of alloxan on the blood glucose concentration was showed in female and male animals.After identical doses of alloxan injected,the blood glucose concentration in female animals increased quickly and its peak was higher than that in male animals.About 70% female dogs were successfully induced to become diabetic models with 40 mg/kg alloxan,corresponding only 40% for male dogs(P0.01).The most optimum dose of alloxan for male dogs was 50 mg/kg,but 3 female dogs(30%) injected with this doses were dead because of higher blood glucose concentration.Similar phenomenons were found in mice and the most optimum dose of alloxan was 200 mg/kg and 250 mg/kg for female and male respectively.Conclusion Sensitivity of alloxan to female is higher than that to male.For male animals,the dose of alloxan used to duplicate diabetic animal models would be higher about 20% than that for female animals.