Objective To prepare CD318-specific monoclonal antibodies and evaluate their specificity, affinity, and application in immunological detection, laying the foundation for the development of CD318-targeted antibody drugs. MethodsCD318 protein was expressed and purified, and was used as an antigen to immunize mice, then mice with higher antiserum titers were screened. We prepared CD318-specific monoclonal antibodies through cell fusion and monoclonal screening, and the specificity, affinity, and application of the obtained monoclonal antibodies in immunological assays were evaluated. Then we constructed a CD318/CD3-targeting bispecific antibody and assessed its impact on T-cell cytotoxicity. Results Thirteen monoclonal antibodies were successfully generated, with the hybridoma clone 13-8-G2 exhibiting the highest titer, strongest specificity, and broadest applicability. The antibody was identified as an IgG1 isotype with a kappa light chain. The variable region of the light chain measured 318 bp, while the heavy chain variable region was 357 bp, yielding an affinity constant of approximately 7.68×10⁹. The specificity of CD318 was confirmed using flow cytometry and immunofluorescence assays. Additionally, a CD318/CD3-targeting bispecific antibody was constructed using the variable regions of this CD318 monoclonal antibody, which demonstrated enhanced T-cell cytotoxicity. Conclusion High-affinity and highly specific CD318 monoclonal antibodies were successfully prepared, laying a foundation for the development of therapeutic antibodies targeting CD318.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice ; Antibodies, Bispecific/immunology* ; Humans ; Mice, Inbred BALB C ; Antibody Specificity/immunology* ; CD3 Complex/immunology* ; Antigens, CD/genetics* ; T-Lymphocytes/immunology* ; Hybridomas/immunology* ; Female

Objective:To observe the clinical efficacy of Tuina(Chinese therapeutic massage)plus electroacupuncture in treating migraine due to liver-Yang hyperactivity and the effects on the serum levels of calcitonin gene-related peptide(CGRP)and prostaglandin(PG)E2. Methods:A total of 122 patients with migraine due to liver-Yang hyperactivity were recruited and randomized into a control group and an observation group,each consisting of 61 cases.The control group was given Tong Nao Huo Luo acupuncture(acupuncture treatment for unblocking brain collaterals),and the observation group was Tuina treatment focusing on cervical Ashi points in addition to the intervention received by the control group.Both groups were treated once daily for 21 consecutive days.When the intervention finished,the two groups were observed for changes in the headache score,symptom and sign scores of traditional Chinese medicine(TCM),the severity of impact on life,and serum CGRP and PGE2 levels.The clinical efficacy was compared after 21 d of treatment. Results:The observation group had a higher total effective rate than the control group,90.2%versus 73.8%(P<0.05);after treatment,the headache and TCM symptom and sign scores decreased in both groups(P<0.05)and were lower in the observation group than in the control group(P<0.05);the migraine's impact on life was less severe in the observation group than in the control group(P<0.05);the levels of serum CGRP and PGE2 dropped in the two groups(P<0.05)and were lower in the observation group(P<0.05). Conclusion:Tong Nao Huo Luo acupuncture can produce more significant efficacy in treating migraine due to liver-Yang hyperactivity when combined with cervical Tuina at Ashi points,better alleviating the headache,improving TCM symptoms and body signs,and reducing the impact of headache on life.The mechanism may be associated with inhibiting the expression of serum pain factors CGRP and PGE2.

OBJECTIVE To investigate whether quinolinic acid(QA)induces autophagy in PC12 cells and its relationship with glycogen synthase kinase-3β(GSK-3β)/β-catenin related signaling path?ways. METHODS PC12 cells were treated with QA 2.5,5.0 and 10.0 mmol·L-1 for 24 h. The cell viability was determined by MTT assay. Autophagy fluorescent spots labelled form of microtubule-associated protein 1 light chain 3(LC3)was examined by LC3 immunostaining. The expressions of GSK-3β,β-catenin,LC3 and Beclin 1 were determined by Western blotting. RESULTS QA inhibited PC12 cell survival in a concentration-dependent manner,and IC50 was 8.7 mmol · L- 1. Compared with normal control group,QA 2.5,5.0 and 10.0 mmol · L-1 increased autophagic intracellular LC3 fluorescence spots,elevated the expression ratio of LC3-Ⅱ/LC3-Ⅰ and expression of Beclin 1 in PC12 cells(P<0.05). In addition,QA enhanced GSK-3βexpression and decreasedβ-catenin expression(P<0.05,P<0.01). CONCLUSION QA induces autophagy in PC12 cells. This mechanism may be associated with the activation of GSK-3β/β-catenin related signaling pathways.