Objective To investigate the safety and clinical response of dendritic cells (DC)vaccine loaded with HLA-A2-restricted peptides MAGE-3、 Tyr、 MART-1 and GP-100 against malignant melanoma. Methods Twenty three HLA-A2 positive patients with malignant melanoma were enrolled.Peripheral blood mononuclear cells were separated into adherent cells for induction of DC loaded with four peptides. DC vaccine was administered subcutaneously once a week in inguinal region. The immunological responses and clinical responses were evaluated. Results Dendritic cell vaccination were well tolerated in all patients and there was no toxicity observed. Serum levels of IL-2, IL-12 and IFN-γincreased significantly after first vaccination and fourth vaccination. The positive rate of DTH test was 50% (5/10), and 2 fold increase of CD8+ IFN-γ+ cells were observed in 6 of 10 patients. Stable disease was observed in 11 of 23 patients, one patient had a complete metastasis regression, four patients had 50% regression of metastasis,four patients had a minor response, and disease progressed in three patients. Conclusion DC vaccines loaded with peptides MAGE-3、 Tyr、 MART-1 and GP-100 can elicit non-specific and specific immune responses, leading disease control. DC vaccine is considered one of safe and effective treatments for malignant melanoma.

Objective To optimize the technical parameters used for embryo transfer in golden hamsters,and explore the optimal number and developmental period of transplanted embryos for golden hamster pregnancy recipients.Methods We established a population of true pregnant albino golden hamster embryo transfer recipients and compared the effects of different embryonic developmental stages,recipient embryo reduction,and secondary transfer recipients on litter yield,donor embryo yield,and offspring survival rate.Results Compared with wild-type golden hamster transplant recipients,true pregnancy albino recipients allowed the origin of the offspring to be determined quickly,and were suitable for various reproductive experimental programs based on embryo transfer.The use of fertilized eggs or two-cell embryos had no significant impact on the transfer effect(P＞0.05);however,the rate of donor embryos was significantly increased in the recipient embryo-reduction group(22％,P＜0.05).The second transfer recipient's non-pregnancy rate was significantly increased(42％,P＜0.01).The highest embryo yield rate(27％,P＜0.01)and normal survival rate(89％)occurred with the transfer of 6～10 embryos.Conclusions The transfer of 6～10 donor embryos may improve the yield and survival rate of donor embryos.Here,we successfully established an embryo transfer method using pregnant albino golden hamsters as recipients,thus providing technical support for the application and development of gene modification models in golden hamsters.

Objective:To preliminarily analyze the expression of angiotensin-converting enzyme 2 (ACE2) and to investigate its potential regulatory mechanism in chronic rhinosinusitis with nasal polyps (CRSwNP).Methods:Patients underwent nasal endoscopic surgery in the Third Affiliated Hospital of Sun Yat-sen University from February 2020 to May 2021 were selected, including 17 males and 6 females, aging from 23 to 66 years old. Expression of ACE2 was evaluated via immunohistochemical staining in controls with non-chronic rhinosinusitis, non-eosinophilic CRSwNP (non-ECRSwNP), and eosinophilic CRSwNP (ECRSwNP) tissue, respectively. Correlations between ACE2 and the indicated Th1/Th2-related cytokines (IFN-γ, IL-4, IL-5, IL-13, IL-25, IL-33, TSLP and periostin) were analyzed based on GSE72713 dataset. Protein-protein interaction (PPI) network was constructed via string database, immune infiltration of GSE72713 dataset was evaluated using cibersort algorithm. ACE2 was comprehensively analyzed by microRNA regulatory network, gene set enrichment analysis (GSEA) and pharmacological analysis. Statistical analysis was performed using GraphPad 7.0 and SPSS 20.0 software.Results:ACE2 was up-regulated in non-ECRSwNP compared with ECRSwNP. Microarray analysis showed that ACE2 was positively correlated with IFN-γ while inversely correlated with IL-5, IL-13 and periostin significantly. Analysis of immune infiltration suggested that ACE2 expression correlated positively with the number of M1 macrophage while negatively with M2 macrophage. GSEA demonstrated that interferon-related signaling pathways were up-regulated in non-ECRSwNP, and miRNA-200B/miRNA-200C/miRNA-429 pathways targeting ACE2 were enriched in ECRSwNP. Results of pharmacological analysis indicated that ampicillin was able to promote the expression of ACE2 whereas acetaminophen could down regulated the expression of ACE2.Conclusion:Expression pattern of ACE2 is varied in non-ECRSwNP and ECRSwNP, which may be related to the different infiltration of indicated cytokines and different regulatory pathways of miRNA.