Japanese encephalitis virus(JEV)belongs to the genus Flavivirus and the family Flavi-viridae,and is classified as a single-stranded,positive-sense RNA virus.The disease known as Japa-nese encephalitis(JE),which results from JEV infection,is a viral zoonosis that is prevalent worldwide and poses a significant public health concern.JEV infection activates a variety of signa-ling pathways,leading to a series of changes that are crucial to the virus's pathogenesis.Among these pathways,Toll-like receptors(TLRs)are particularly significant,and their diverse range and complex signal transduction mechanisms present substantial challenges for the prevention and con-trol of JEV.Currently,there is no specific treatment for JEV.Although some vaccines have been developed to prevent JE,eradicating JEV remains difficult due to its zoonotic transmission cycle and the limited efficacy of the available vaccines.This article reviews the alterations in various TLR signaling pathways induced by JEV infection in the host,aiming to provide insights into the patho-genic mechanisms of JEV and to identify potential new antiviral targets.

Porcine reproductive and respiratory syndrome virus(PRRSV)is the pathogen of porcine reproductive and respiratory syndrome(PRRS),and its infection mainly causes abortion,stillbirth in sows and piglet respiratory infections,which are widely prevalent in the world and seriously jeopardize the development of the world's animal husbandry industry.PRRSV infection of the host is capable of inducing significant immunosuppression,and in recent years,the study of the mecha-nism of PRRSV immunosuppression has become a hot topic,with studies showing that numerous PRRSV proteins are involved in the regulation of host innate immunity and elucidating the mecha-nism by which PRRSV proteins modulate host innate immunity.In this paper,we reviewed the progress of research on the interaction mechanism between PRRSV proteins and host innate im-munity to provide a theoretical basis for a comprehensive understanding of the pathogenesis of PRRSV and the prevention and control of PRRS.

Japanese encephalitis virus(JEV)belongs to the genus Flavivirus and the family Flavi-viridae,and is classified as a single-stranded,positive-sense RNA virus.The disease known as Japa-nese encephalitis(JE),which results from JEV infection,is a viral zoonosis that is prevalent worldwide and poses a significant public health concern.JEV infection activates a variety of signa-ling pathways,leading to a series of changes that are crucial to the virus's pathogenesis.Among these pathways,Toll-like receptors(TLRs)are particularly significant,and their diverse range and complex signal transduction mechanisms present substantial challenges for the prevention and con-trol of JEV.Currently,there is no specific treatment for JEV.Although some vaccines have been developed to prevent JE,eradicating JEV remains difficult due to its zoonotic transmission cycle and the limited efficacy of the available vaccines.This article reviews the alterations in various TLR signaling pathways induced by JEV infection in the host,aiming to provide insights into the patho-genic mechanisms of JEV and to identify potential new antiviral targets.

Porcine reproductive and respiratory syndrome virus(PRRSV)is the pathogen of porcine reproductive and respiratory syndrome(PRRS),and its infection mainly causes abortion,stillbirth in sows and piglet respiratory infections,which are widely prevalent in the world and seriously jeopardize the development of the world's animal husbandry industry.PRRSV infection of the host is capable of inducing significant immunosuppression,and in recent years,the study of the mecha-nism of PRRSV immunosuppression has become a hot topic,with studies showing that numerous PRRSV proteins are involved in the regulation of host innate immunity and elucidating the mecha-nism by which PRRSV proteins modulate host innate immunity.In this paper,we reviewed the progress of research on the interaction mechanism between PRRSV proteins and host innate im-munity to provide a theoretical basis for a comprehensive understanding of the pathogenesis of PRRSV and the prevention and control of PRRS.

Porcine deltacoronavirus(PDCoV)is the main pathogen of porcine deltacoronavirus dis-ease.After infection,pigsmanifest a series of main symptoms,such as persistent vomiting,watery diarrhea and severe dehydration.Pigs at almost all growth stages are likely to be infected with the virus,especially suckling piglets are much sensitive to the virus.Once PDCoV infects the host,it u-sually causes significant immunosuppression.In recent years,studies on the immunosuppressive mechanism of PDCoV have gradually attracted widespread attention.The results showed that mul-tiple proteins of PDCoV were involved in the regulation of host innate immunity,revealing the mechanism of these proteins in regulating host innate immunity.In this paper,the interaction mechanism between PDCoV protein and host innate immunity were rsummarized,which will pro-vide a theoretical basis for further understanding the pathogenesis of PDCoV and effective preven-tion and control of porcine delta coronavirus disease.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.

Porcine deltacoronavirus(PDCoV)is the main pathogen of porcine deltacoronavirus dis-ease.After infection,pigsmanifest a series of main symptoms,such as persistent vomiting,watery diarrhea and severe dehydration.Pigs at almost all growth stages are likely to be infected with the virus,especially suckling piglets are much sensitive to the virus.Once PDCoV infects the host,it u-sually causes significant immunosuppression.In recent years,studies on the immunosuppressive mechanism of PDCoV have gradually attracted widespread attention.The results showed that mul-tiple proteins of PDCoV were involved in the regulation of host innate immunity,revealing the mechanism of these proteins in regulating host innate immunity.In this paper,the interaction mechanism between PDCoV protein and host innate immunity were rsummarized,which will pro-vide a theoretical basis for further understanding the pathogenesis of PDCoV and effective preven-tion and control of porcine delta coronavirus disease.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.

Objective To analyze the serum levels of bone morphogenetic protein 4(BMP4)and nerve ax-on guidance factor 4(Netrin-4)in patients with diabetic retinopathy(DR),and to study their relationship with disease stage and prognosis.Methods A total of 186 patients with type 2 diabetes admitted to the hospi-tal from October 2019 to November 2022 were selected as the research objects.According to the presence or absence of retinopathy,they were divided into DR Group(108 cases)and non-DR group(78 cases),and 100 healthy people in the hospital were selected as the control group.According to the stage of DR,the DR pa-tients were divided into stage Ⅰ(19 cases),stage Ⅱ(14 cases),stage Ⅲ(22 cases),stage Ⅳ(23 cases),stage V(18 cases)and stage Ⅵ(12 cases).According to the visual impairment after treatment,the DR patients were divided into a good prognosis group(72 cases)and a poor prognosis group(36 cases).Serum BMP4,Ne-trin-4,fasting plasma glucose(FPG),glycated hemoglobin Alc(HbA1c),systolic blood pressure(SBP),dias-tolic blood pressure(DBP),triglyceride(TG),total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)were detected by enzyme-linked immunosorbent as-say.The serum levels of BMP4 and Netrin-4 in DR patients with different disease stages were analyzed.Multi-variate Logistic regression was used to analyze the influencing factors of DR,and the serum levels of BMP4 and Netrin-4 in DR patients with different prognosis were analyzed.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of poor prognosis in DR patients.Results The FPG,HbA1c,SBP,DBP,TG,TC,BMP4,Netrin-4 levels in DR group were higher than those in control group and non-DR group,while HDL-C level was lower than that in control group and DR group(P＜0.05).There were significant differences in serum BMP4 and Netrin-4 levels between DR patients with different disease stages(P＜0.05).HbA1c,BMP4 and Netrin-4 levels were influencing factors for the occurrence of DR(P＜0.05).The serum levels of BMP4 and Netrin-4 in the poor prognosis group were higher than those in the good prog-nosis group(P＜0.05).The combination of serum BMP4 and Netrin-4 levels in the diagnosis of poor progno-sis of DR patients was better than that of each index alone.Conclusion The serum levels of BMP4 and Ne-trin-4 are increased in DR patients,which can assist in the evaluation of the disease stage of the patients.The combination of BMP4 and Netrin-4 has a good effect in the diagnosis of poor prognosis of patients.

Objective:To investigate the combined therapy of calcium dobesilate capsules, oral administration of Dahuoluo capsules, and intravenous drip of Danhong injection in treatment of patients with nonproliferative diabetic retinopathy.Methods:106 cases were randomly selected from the patients with nonproliferative diabetic retinopathy admitted in Traditional Chinese and Western Hospital of Dazhou City from Jan. 2019 to Dec. 2019. Based on the random numbers table, the cases were divided into control group and intervention group, with each group 53 patients. They were respectively treated with the therapy of calcium dobesilate capsules alone and the combined therapy of calcium dobesilate capsules, oral administration of Dahuoluo capsules and intravenous drip of Danhong injection, and the therapeutic effects were compared.Results:After treatment, the visual acuity of patients in both groups significantly improved ( P<0.05) , and the visual acuity of patients in the intervention group was higher than that of patients in the control group ( P<0.05) ; after treatment, the peak systolic velocity (PSV) and end diastolic velocity (EDV) of the central retinal artery (CRA) and posterior ciliary artery (PCA) of the patients in both groups significantly increased ( P<0.05) , while RI significantly reduced ( P<0.05) , and PSV and EDV of CRA and PCA in the intervention group were significantly higher than those of the control group ( P<0.05) , while RI was significantly lower than that of the control group ( P<0.05) . After treatment, the therapeutic efficiency of the intervention group was 96.23%, significantly higher than 79.25% in the control group ( χ2=7.102, P<0.05) . The incidence of adverse reactions in the intervention group was 9.43%, which had no significant difference with 3.78% in the control group ( χ2=1.377, P>0.05) . After targeted treatment was given, the adverse reactions disappeared. Conclusion:The combined therapy of calcium dobesilate capsules, oral administration of Dahuoluo capsules, and intravenous drip of Danhong injection was significantly effective in treatment of patients with nonproliferative diabetic retinopathy.