Objective: This study aimed to provide clinical evidence for lineage replacement and genetic changes of High-Risk Human Papillomavirus (HR-HPV) during the period of vaccine coverage and characterize those changes in eastern China. Methods: This study consisted of two stages. A total of 90,583 patients visiting the Obstetrics and Gynecology Hospital of Fudan University from March 2018 to March 2022 were included in the HPV typing analysis. Another 1076 patients who tested positive for HPV31, 33, 52, or 58 from November 2020 to August 2023 were further included for HPV sequencing. Vaccination records, especially vaccine types and the third dose administration time, medical history, and cervical cytology samples were collected. Viral DNA sequencing was then conducted, followed by phylogenetic analysis and sequence alignment. Results: The overall proportion of HPV31 and 58 infections increased by 1.23% and 0.51%, respectively, while infection by HPV33 and 52 decreased by 0.42% and 1.43%, respectively, within the four-year vaccination coverage period. The proportion of HPV31 C lineage infections showed a 22.17% increase in the vaccinated group, while that of the HPV58 A2 sublineage showed a 12.96% increase. T267A and T274N in the F-G loop of HPV31 L1 protein, L150F in the D-E loop, and T375N in the H-I loop of HPV58 L1 protein were identified as high-frequency escape-related mutations. Conclusions: Differences in epidemic lineage changes and dominant mutation accumulation may result in a proportional difference in trends of HPV infection. New epidemic lineages and high-frequency escape-related mutations should be noted during the vaccine coverage period, and regional epidemic variants should be considered during the development of next-generation vaccines.

Type VII collagen (Col VII) is a key component of anchoring fibrils that stabilize the dermal-epidermal junction. To elucidate its functional properties and evaluate its biosafety, we designed several constructs C7PX (X = 1-16) of recombinant humanized collagen VII (rhCol VII) for expression in Escherichia coli. Among these, C7P7 exhibited the highest yield (69.03 mg/L), and its circular dichroism spectrum and differential scanning calorimetry analysis displayed characteristic features of a triple-helix structure. Thus, C7P7 was selected as the final rhCol VII candidate for subsequent analysis of physicochemical properties, biosafety, and functional performance. Comprehensive characterization-including peptide coverage analysis, N- and C-terminal amino acid sequencing and molecular weight-confirmed that the rhCol VII possessed an amino acid sequence fully consistent with its theoretical design. Biocompatibility testing, performed in accordance with national standard GB/T 16886, equivalent to ISO 10993, demonstrated excellent biosafety of rhCol VII. In vivo fluorescence imaging of Cy5-labeled rhCol VII showed specific localization to skin tissue following subcutaneous injection, with negligible distribution to major internal organs (heart, liver, spleen, lung, and kidney) at 48 h post-injection. Functionally, rhCol VII markedly enhanced fibroblast proliferation and accelerated wound closure in vivo, confirming its potent biological activity. Collectively, these findings establish rhCol VII as a safe, bioactive macromolecule with promising regenerative potential. Its high production yield, preserved triple-helical structure, and favorable biocompatibility support potential for further development for clinical applications in regenerative medicine.

Objectives: Understanding the antibody response in monkeypox virus (MPXV)-infected and uninfected individuals is essential for developing next-generation MPXV vaccines. This study aimed to characterize neutralizing antibody (NAb) and antibody-dependent cellular cytotoxicity (ADCC) responses in both groups, providing insights into immune protection and vaccine design. Methods: A recombinant vaccinia Tian Tan (VTT) virus was utilized to develop high-throughput luciferase-reporter-based neutralization and ADCC assays. These assays were applied to evaluate the presence and levels of poxvirus-specific antibodies in MPXV-infected and uninfected individuals, including those vaccinated with vaccinia-based vaccines. Results: Poxvirus-specific NAbs were detected in MPXV-negative individuals with prior vaccinia vaccination. However, MSM individuals exhibited significantly lower pre-existing NAb levels than non-MSM individuals, potentially contributing to their higher susceptibility to MPXV infection. In individuals with mild MPXV infection, robust NAb and ADCC responses were observed, regardless of vaccination status. Additionally, HIV-positive individuals demonstrated comparable antibody responses following MPXV infection. Conclusions: These findings highlight the potential role of pre-existing NAbs in MPXV susceptibility and the strong immune response elicited by mild MPXV infection. Further research is needed to determine whether MPXV-specific antibodies mitigate disease progression, which could inform the development of effective MPXV vaccines.