To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

To establish a highly specific,sensitive,and efficient method for detecting antibodies a-gainst the Erns protein of classical swine fever virus(CSFV),and to distinguish CSFV vaccine strains from wild strains infections in combination with the E2 subunit vaccine.The purified Erns protein of the CSFV expressed by baculovirus was conjugated to carboxylated magnetic beads as a solid-phase carrier and horseradish peroxidase(HRP),separately.A double-antigen sandwich chemiluminescent enzyme immunoassay(CLEIA)was developed by optimizing various reaction parameters using a fully automated chemiluminescence analyzer.This method was then applied to quantitatively detect Erns protein antibodies in sera from pigs infected with prevalent strains and those immunized with the CSFV E2 subunit vaccine and challenged with field strains.The results showed that the optimal conditions for coupling protein-to-magnetic bead were as follows:coupling buffer pH of 8.0,a protein coupling amount of 2.5 mg/g,blocking solution of 10％BSA,serum sample volume of 20 μL.The optimal dilution of enzyme-labeled antigen was at 1:500 with a one-step reaction time of 15 minutes.The cutoff value of the established CLEIA method for detecting CSFV Erns protein antibodies was 5.83 U/mL and a diagnostic sensitivity of 1:128.No cross-reac-tivity was observed with positive sera against African swine fever virus,pseudorabies virus,porcine circovirus type 2,porcine epidemic diarrhea virus,porcine reproductive and respiratory syndrome virus,or porcine gastroenteritis virus.Additionally,the method yielded negative results with sera from pigs immunized with the E2 subunit vaccine.In repeatability tests,the intra-assay coefficient of variation(CV)ranged from 0.77％to 11.56％,and the inter-assay CV ranged from 10.30％to 14.55％,both below 15％.The positive and negative concordance rates with a commercial CSFV Erns protein antibody detection kit were 95.24％and 92.71％,separately,with an overall concord-ance rate of 93.23％.The double-antigen sandwich chemiluminescence method established in this study exhibits high sensitivity,excellent repeatability,and suitability for automated detection,making it applicable for serological differentiation between CSFV E2 subunit vaccination and infec-tion with prevalent strains.

N1-methyladenosine(m1A)RNA methylation is critical for regulating mRNA translation;however,its role in the development,progression,and immunotherapy response of head and neck squamous cell carcinoma(HNSCC)remains largely unknown.Using Tgfbr1 and Pten conditional knockout(2cKO)mice,we found the neoplastic transformation of oral mucosa was accompanied by increased m1A modification levels.Analysis of m1A-associated genes identified TRMT61A as a key m1A writer linked to cancer progression and poor prognosis.Mechanistically,TRMT61A-mediated tRNA-m1A modification promotes MYC protein synthesis,upregulating programmed death-ligand 1(PD-L1)expression.Moreover,m1A modification levels were also elevated in tumors treated with oncolytic herpes simplex virus(oHSV),contributing to reactive PD-L1 upregulation.Therapeutic m1A inhibition sustained oHSV-induced antitumor immunity and reduced tumor growth,representing a promising strategy to alleviate resistance.These findings indicate that m1A inhibition can prevent immune escape after oHSV therapy by reducing PD-L1 expression,providing a mutually reinforcing combination immunotherapy approach.

BACKGROUND:Interleukin-1 receptor-associated kinase 4 activity-induced inflammations and infection have been extensively accepted. However, there was no report concerning its effects on flap ischemia-reperfusion injury. OBJECTIVE:To explore the significance of interleukin-1 receptor-associated kinase 4 activity in flap ischemia-reperfusion injury. METHODS:A total of 36 adult male Sprague-Dawley rats were randomized into sham-operated group (n=12), ischemia-reperfusion group (n=12) and interleukin-1 receptor-associated kinase 4 group (n=12). The models of right lower abdominal island flap ischemia-reperfusion injury were set up. Interleukin-1 receptor-associated kinase 4 group was intraperitoneal y injected with 1 mL of interleukin-1 receptor-associated kinase 4 (100μmol/L) before reperfusion. The flaps were col ected at 1, 2, 4, and 6 hours after ischemia-reperfusion injury for histopathhological observation. At 1 hour after ischemia-reperfusion, protein expression of interleukin-1 receptor-associated kinase 4 was detected in flaps. The proportion of flap survival was calculated at 7 days after surgery.RESULTS AND CONCLUSION:Histopathological observation demonstrated that compared with the ischemia-reperfusion injury group, neutrophil infiltration and edema was evidently improved, and the protein expression of interleukin-1 receptor-associated kinase 4 was gradual y reduced in the interleukin-1 receptor-associated kinase 4 group. Flap survival proportions were respectively (51.70 ±7.62)%and (86.56±12.23)%in the ischemia-reperfusion injury group and interleukin-1 receptor-associated kinase 4 group at 7 days after surgery. There were significant differences in the flap survival proportion between the two groups (P<0.01). These results showed that after flap ischemia-reperfusion injury, the inhibition of interleukin-1 receptor-associated kinase 4 activities could elevate the survival rate of transplanted flap.

Objective To investigate the method and effect of free autogenous palmaris longus tendon transplantation in the treatment of moderate and severe cicatricial ectropion.Methods The autogenous palmaris longus tendon was obtained through a small lateral proximal wrist band incision (lengths from 4 cm to 6 cm).The graft was properly sutured to the exposed tarsus and the attachments of inner and outer canthus.Local skin flap was rotated to cover the tendon and to repair the wound.Results Six man (8 eyes) with moderate and severe cicatricial ectropion were treated in our department,all case of ectropion were rectified after operation.The eyeballs were well contacted with lower eyelid and no recurrence of eetropion,epiphora and corneal exposure during 6 to 18 months' follow-up.The tension of the lower eyelid was abiding with satisfied effects.Conclusions The palmaris longus tendon is easy to obtain and there is no harm to the donor site.The cicatricial ectropion could be repaired with self-palmaris longus tendon transplantation with lasting effect and the recurrence rate is lower,which is well worthy popularizing.