Cell & Bioscience

BackgroundDespite chemo-immunotherapy has been applied to the neoadjuvant treatment of non-small cell lung cancer (NSCLC), the impacts of dosage and the order of medication on treatment efficacy and safety remain largely unexplored. We originally designed an exploratory study to investigate the efficacy and safety of reduced-dose chemotherapy combined with delayed immunotherapy as well as the dynamic changes of circulating tumor DNA (ctDNA) and T cell receptor (TCR) during the therapy. MethodsPatients with clinical stage IIA to IIIA resectable NSCLC were treated with 2 cycles of reduced-dose platinum-based chemotherapy on day 1 combined with immunotherapy on day 5. The same postoperative modified adjuvant therapy regimen was administered for 2 cycles. Plasma samples at different time-points were collected and performed with T cell receptor (TCR) and circulating tumor DNA (ctDNA) sequencing. Results38 patients received modified chemo-immunotherapy. The proportion of patients exhibiting complete response and partial response was 5.3% and 68.4%, respectively. The confirmed objective response rate was 73.7%. Radiological downstaging was achieved in 39.5%. Major pathologic response and complete pathologic response were observed in 47.4% and 31.6% of patients, respectively. Only one patient experienced grade 3 adverse event. Further analyses revealed that this modified chemo-immunotherapy led to the expansion of predominant TCR clones and reduction of tumor burden after the first cycle of chemotherapy. ConclusionThe promising clinical efficacy and low side effects of modified neoadjuvant chemo-immunotherapy position it as a prospective and innovative strategy for NSCLC. Trial registration: Registration Number: ChiCTR2000033092

Background: Human papillomaviruses (HPVs) pose a severe threat to global public health by driving nonmelanoma skin cancer (NMSC) and cervical cancer, with NMSC being one of the most common cancers worldwide. Epidermodysplasia verruciformis (EV) is an inborn error of immunity characterized by an increased susceptibility to persistent infection of cutaneous HPV and a high risk of NMSC. The genetic basis remains unknown in many patients with EV. Methods: We collected four unrelated pedigrees with EV. Genetic analysis identified five variants in JAK1 encoding the Janus kinase 1. Ex vivo models and patient-derived tissue were employed to evaluate the functional effects of JAK1 variants and delineate the pathogenic mechanisms. Results: We identified different variants in JAK1 in four pedigrees with dominant EV. Genetic analysis revealed five novel variants in JAK1, three of which resulted in nonsense-mediated mRNA decay (NMD). Functional assays identified a decreased phosphorylation of the signal transducers and activators of transcription (STATs), impaired interferon responses, and defective T cell activation. Immune dysregulation in patients, characterized by a reduced CD4/CD8 T cell ratio, decreased CD8 naive T cell proportion, and accumulated memory T cells, implies impaired antiviral immunity against HPV. Conclusions: Our findings confirm that JAK1 loss-of-function (LOF) variants underlie susceptibility to cutaneous HPV infection. [Funded by the National Natural Science Foundation of China (81788101, 81230015, 82394420, and 82394423), the National Key Research and Development Program of China (2022YFC2703900), the CAMS Innovation Fund for Medical Sciences (2021-I2M-1-018), and the Regione Lombardia, Italy (Innovative Research Project 1137-2010)].

BackgroundPrimary central nervous system lymphoma (PCNSL) is clinically challenging due to its location and small biopsy size, leading to a lack of comprehensive molecular and biologic description. We previously demonstrated that 91% of PCNSL belong to the activated B-cell-like (ABC) molecular subtype of diffuse large B-cell lymphoma (DLBCL). MethodsHere we investigated the expression of 739 cancer related genes in HIV(-) patients in 25 ABC-PCNSL and 43 ABC-systemic DLBCL, all tumors were EBV(-). ResultsWe found 135 genes which were identified as differentially expressed between these ABC-PCNSL and ABC-systemic DLBCL (p<0.05). The ABC-PCNSL showed higher gene expression in several cancer-related gene sets including genes related to Hedgehog, DNA damage repair, Wnt and MAPK signaling. Whole exome sequencing showed distinct genetic features of PCNSL compared to DLBCL, CXCR4 mutations in particular, that have been associated with ibrutinib resistance. In a focused analysis, PCNSL and DLBCL cases that fall into the "MCD" genetic subtype showed substantial overlap. These data provide detailed information about unique characteristics of PCNSL in HIV(-) patients as compared to comparable DLBCL subtypes.

Papillary thyroid cancer (PTC) is the most common thyroid malignancy. Although PTC usually has a favorable prognosis, some aggressive PTC subtypes and lymph node (LN) metastasis contribute to high rates of recurrence and poor clinical outcomes. We analyzed single-cell RNA sequencing (scRNA-seq) data from 15 samples, including primary tumors of PTC, metastatic LNs, and paracancerous tissues. After quality filtering, 28,205 cells were detected. Of these, 13,390 cells originated from 7 tumor tissues, 2,869 cells from 2 metastatic LNs, and 11,945 cells from 6 paracancerous tissues. The increase in the proportion of CD4+ Tregs may be a key factor responsible for the immunosuppressive property of PTC. A novel cell type was identified, named Protective EGR1+CD4+ T cell, which might be antagonistic to the CD4+ Tregs and inhibit the formation of the immunosuppressive microenvironment and tumor immune evasion. Inhibitory checkpoints TIGIT and CD96 were found to be better targets than PD-1 for immune therapy in PTC patients with LN metastasis. For PTC patients without LN metastasis, however, PD-1, TIGIT, and CD96 could be suitable targets of immunotherapy. These findings would contribute to the further understanding of molecular mechanisms resulting in occurrence and development of PTC, and provide a theoretical rationale for targeted therapy and immunotherapy.

Background and PurposeThe liver cancer (LC) is a highly malignant tumor of the digestive system with a poor prognosis. Cuproptosis is a new type of regulated cell death that has been found by researchers. The expression of cuproptosis-related genes in LC and their relevance to prognosis, on the other hand, remain unknown. This study aimed to explore a gene signature to predict the liver cancer prognosis and identified the vital gene. Experimental approachThe expression patterns of RNA and related clinical data of 371 LC patients were obtained based on The Cancer Genome Atlas (TCGA). Differentially expressed genes (DEGs) were acquired by comparing tumors with adjacent normal samples. Genes displaying significant association with OS were screened through univariate Cox regression analysis and the least absolute shrinkage and selection operator (LASSO) algorithm. All cases were classified into the validation or training group to validate the constructed gene signature. We conducted real-time polymerase chain reaction (PCR) and assays for transwell invasion, CCK-8, and colony formation to determine the biological roles of DLAT. Key ResultsThe differential expression of twelve cuproptosis regulators in LC and normal liver tissues was discovered in this investigation. DEGs can be used to distinguish between two forms of LC. Cuproptosis-related genes were evaluated for survival predictive significance using the Cancer Genome Atlas (TCGA) cohort. A 3-gene signature based on least absolute shrinkage and selection operator (LASSO) Cox regression was used to categorize an LC patient cohort from the TCGA into low- and high-risk categories. Patients in the low-risk group had a considerably higher likelihood of surviving (P = 0.05) than those in the high-risk group. When paired with clinical parameters, risk score was an independent predictor in predicting the OS of patients with LC. Conclusions & ImplicationsCuproptosis-related genes thus play an important role in tumor formation and can be used to predict the prognosis of LC patients. DLAT has the best prognostic value and can be a therapeutic target for liver cancer.

In this study, we report the development of a tuberculosis meningitis (TBM) mouse model using the HN878 strain via the aerosol route. Three genetically different strains of mice, CB6F1, C57BL/6 and BALB/c mice, were used to determine the dissemination of HN878 in the brain. The 8 x 108 CFU/ml of HN878 dose was used to infect CB6F1 mice and deposited approximately 3.8 (0.07, SD) log10 CFU in the lungs. The burden of HN878 in the brain of the control group (administered saline) after approximately 16/17 days post-infection for CB6F1, C57BL/6 and BALB/c were 4.00 (0.47 SD), 3.79 (0.27, SD) and 2.12 (0.41, SD) log10 CFU/brain, respectively. The log10 CFU/brain in the BCG vaccinated CB6F1, C57BL/6 and BALB/c mice were 1.05 (0.61, SD), 2.13 (0.33, SD) and 1.42 (0.38, SD) respectively, which, if compared to the control groups, BCG vaccinated mice inhibited dissemination of HN878 in the brain by an impressive 2.94 (CB6F1), 1.66 (C57BL/6) and 0.69 (BALB/c) log10 CFU/brain reduction. In conclusion, we have established a relatively inexpensive TBM mouse model using an aerosol, a natural route of infection, which will advance research in understanding TBM dissemination to the brain, and preclinical tuberculosis vaccine/drug discovery/drug regimens against TBM.

BackgroundCurrently, no biomarkers are available to identify resistance to androgen-deprivation therapies (ADT) in men with hormone-naive prostate cancer. Since 5-hydroxymethylcytosines (5hmC) in gene body are associated with gene activation, in this study, we evaluated whether 5hmC signatures in cell-free DNA (cfDNA) predicts early resistance to ADT. ResultsWe collected a total of 139 serial plasma samples from 55 prostate cancer patients receiving ADT at three time points including baseline (prior to initiating ADT, N=55), 3-month (after initiating ADT, N=55), and disease progression (N=15) within 24 months or 24-month if no progression was detected (N=14). To quantify 5hmC abundance across the genome, we used selective chemical labeling sequencing and mapped sequence reads to individual genes. Differential methylation analysis in baseline samples identified significant 5hmC difference in 1,642 of 23,433 genes between patients with and without progression (false discovery rate, FDR<0.1). Patients with disease progression showed significant 5hmC enrichments in multiple hallmark gene sets with androgen responses as top enriched gene set (FDR=1.19E-13). Interestingly, this enrichment was driven by a subgroup of patients featuring a significant 5hmC hypermethylation in the gene sets involving AR, FOXA1 and GRHL2. To quantify overall activities of these gene sets, we developed a gene set activity scoring algorithm and observed significant association of high activity scores with poor progression-free survival (P<0.05). Longitudinal analysis showed that the high activity scores were significantly reduced after 3-months of initiating ADT (P<0.0001) but returned to higher levels when the disease was progressed (P<0.05). ConclusionsThis study demonstrates that 5hmC-based activity scores from gene sets involved in AR, FOXA1 and GRHL2 may be used as biomarkers to determine early treatment resistance, monitor disease progression, and potentially identify patients who would benefit from upfront treatment intensification.

BackgroundDespite the improved survival observed in PD-1/PD-L1 blockade therapy, there still is a lack of response to the anti-PD1 therapy for a large proportion of cancer patients across multiple indications, including non-small cell lung cancer (NSCLC) MethodsTranscriptomic profiling was performed on 57 whole blood samples from 31 NSCLC patients and 5 healthy donors, including both responders and non-responders received anti-PD-1 Tislelizumab plus chemotherapy, to characterize differentially expressed genes (DEGs), signature pathways, and immune cell subsets regulated during treatment. Mutations of oncogenic drivers were identified and associated with therapeutic outcomes in a validation cohort with 1661 cancer patients. These multi-level biomarkers were validated and compared across different methods, external datasets and multiple computational tools. ResultsNSCLC patients examined and achieved pathological complete response (pCR) were considered as responders or non-responders otherwise. Expression of hundreds DEGs (FDR p<0.05, fold change<-2 or >2) was changed in blood during neoadjuvant anti-PD-1 treatment, as well as in lung cancer tissue as compared to normal samples. Enriched PD-1-mediated pathways and elevated cell abundances of CD8 T cells and regulatory T cells were exclusively observed in responder blood samples. In an independent validation cohort of 1661 pan-cancer patients, a panel of 4 top ranked genetic alterations (PTCH1, DNMT3A, PTPRS, JAK2) identified from responders in discovery cohort were found positively associated with the overall survival (p<0.05). ConclusionThese findings suggest peripheral blood-based biomarkers and cell subsets could be utilized to define the response to neoadjuvant PD-1 blockade in NSCLC patients and a set of novel gene mutations is strongly associated with the therapeutic outcome of cancer immunotherapy.

In recent years, chimeric antigen receptor (CAR) T cell therapy has shown great promise in treating hematological malignancies. However, using virus in manufacture of CAR T cells brings about several problems. The application of CRISPR/Cas9 genome editing technology emerges in constructing novel CAR T cells by disrupting endogenous genes. Here we successfully develop a two-in-one approach to generate non-viral genome specific targeted CAR T cells through CRISPR/Cas9. By targeting a CAR in AAVS1 safe harbor locus, we demonstrated that these CAR T cells behave comparable to those conventionally produced by lentivirus. Furthermore, PD1-knockin anti-CD19 CAR T cells show a superior ability to eradicate tumor cells with high PD-L1 expression. In the adoptive therapy for relapsed/refractory (r/r) aggressive B-cell non-Hodgkin lymphoma (B-NHL), we observed durable responses without serious adverse events and complete remission (CR) in patients treated with these PD1 knockout CAR T cells. Collectively, our results prove the safety and feasibility of non-viral genome specific integrated CAR T cells, thus providing a new potential strategy for cancer treatment using these novel CAR T cells.

BackgroundN4-acetylcytidine (ac4C) modification plays a critical role in cancer development. Exploring ac4C modification in laryngeal squamous cell carcinoma (LSCC) may help elucidate its pathogenesis. MethodsLSCC-related datasets were obtained from GEO. After preprocessing and annotating single-cell data, malignant cells were identified by CNV scoring and further divided into subpopulations. Malignant epithelial cells (MECs) were identified and subclustered based on ac4C-related gene activity. Prognostic genes were screened using Cox regression and machine-learning approaches, followed by validation in clinical samples using qPCR. The biological and immunological relevance of these genes was further explored through immune infiltration, immunotherapy response, and mutation analyses. ResultsThe 14,465 identified MECs were classified into five subgroups (MEC1-5), among which MEC3 showed the strongest association with the ac4C gene set. Machine-learning analysis of MEC3-derived genes yielded seven prognostic markers, including BARX1, FHL2, NXPH4, PKMYT1, TNFAIP8L1, CRLF1, and CENPP. qPCR confirmed their differential expression between tumor and adjacent normal tissues. These genes were significantly associated with alterations in the tumor immune microenvironment, with high-risk patients showing increased immune infiltration and immune activity. ConclusionSeven ac4C-related prognostic genes were identified that may contribute to LSCC progression by modulating the tumor immune microenvironment, providing potential therapeutic insights.

BackgroundChronic airway diseases like cystic fibrosis (CF) and primary ciliary dyskinesia (PCD) pose substantial clinical challenges. This study explores the p.C97W variant in WFDC2, proposed as a new genetic origin of respiratory distress, especially among Koreans. MethodsWhole-exome/genome sequencing (WES/WGS) were performed on 64 patients from 62 families presenting with severe bronchiectasis and chronic rhinosinusitis. In vitro analyses and protein modeling were utilized to evaluate the functional implications of the WFDC2 variant. ResultsPathogenic variants were found in 11.3% of families, including a novel homozygous WFDC2 missense variant (c.291C>G, p.Cys97Trp) in five unrelated families, confirmed by Sanger sequencing. The variant, rare globally but more frequent in Koreans, caused persistent wet cough, chronic rhinosinusitis, and bronchiectasis in affected patients. Lung tissue pathology showed chronic inflammation and interstitial fibrosis. The p.C97W variant impaired WFDC2 protein folding, secretion, and function. ConclusionsThe p.C97W variant in WFDC2 is a critical genetic factor in severe chronic airway disease that shares clinical features with CF and PCD. Given its implications for diagnosis and treatment, genetic testing for WFDC2 mutations in individuals with CF or PCD-like symptoms is recommended.

Human pluripotent stem cell (hPSC)-derived brain organoids offer an unprecedented opportunity for various applications as an in vitro model, such as modeling virus infection and drug screening. In this study, we present an experimental brain organoid platform for modeling infection with multiple viruses (e.g., influenza virus or enterovirus). Brain organoids challenged by influenza viruses (H1N1-WSN and H3N2-HKT68) had decreased overall organoid size, similar to ZIKA virus infection, while enteroviruses (EV68 and EV71) infected brain organoids displayed the opposite result. Then, we studied the molecular events in WSN-infected organoids, and we found that WSN could widely infect multiple cell types, and preferentially infected MAP2+ neurons compared to SOX2+ neural stem cells (NSCs) and GFAP+ astrocytes in brain organoids, and induced apoptosis of NSCs and neurons, but not astrocytes. The inflammatory responses in organoids observed to occur (Tumor necrosis factor alpha, interferon gamma, and interleukin 6) after WSN infection may further facilitate brain damage. Furthermore, transcriptional profiling revealed several upregulated genes (CSAG3 and OAS2) and downregulated genes (CDC20B, KCNJ13, OTX2-AS1, CROCC2, and F5) after WSN infection for 24 hpi and 96 hpi, implicating antiviral drugs development responses to WSN. Finally, we explored neurotrophic factors (e.g., BDNF, GDNF, and NT3) and PYC-12 as antiviral and neuroprotective reagents, which could significantly suppress virus infection, apoptosis, and inflammatory responses. Collectively, we established a tractable experimental model system to investigate the impact and mechanism of virus infection on human brain development, and provide a platform for rapidly screening therapeutic compounds, advancing the development of antiviral strategies.

ImportancePrimary biliary cholangitis (PBC) is a classical autoimmune disease, which is highly influenced by genetic determinants. Many genome-wide association studies (GWAS) have reported that numerous genetic loci were significantly associated with PBC susceptibility. However, the effects of genetic determinants on liver cells and its immune microenvironment for PBC remain unclear. ObjectiveTo identify genetics-modulated functional liver cell subsets involved in the pathogenesis of PBC. Design, Setting, and ParticipantsIn this present study, 13,239 European participants were collected from IEU open GWAS project on PBC. There were 1,124,241 qualified SNPs used for GWAS analysis. Expression quantitative trait loci (eQTL) data across 49 tissues were downloaded from the GTEx database. Two single cell RNA sequencing (scRNA-seq) profiles and two bulk-based RNA transcriptomes were downloaded from the GEO database. Data collection and analyses were performed from August 2020 to June 2021. Main outcomes and measuresWe constructed a powerful computational framework to integrate GWAS summary statistics with scRNA-seq data to uncover genetics-modulated liver cell subpopulations. ResultsBased on our multi-omics integrative analysis, we found that 29 risk genes including ORMDL3, GSNK2B, and DDAH2 were significantly associated with PBC susceptibility. Gene-property analysis revealed that four immune cell types, including Cst3+ dendritic cell, Chil3+ macrophage, Trbc2+ T cell, and Gzma+ T cell, were significantly enriched by PBC-risk genes. By combining GWAS summary statistics with scRNA-seq data, we found that cholangiocytes exhibited a notable enrichment by PBC-related genetic association signals (Permuted P < 0.05). The risk gene of ORMDL3 showed the highest expression proportion in cholangiocytes than other liver cells (22.38%). Compared with ORMDL3+ cholangiocytes, there were 71 significantly highly-expressed genes among ORMDL3- cholangiocytes (FDR < 0.05), such as inflammatory cytokine genes CXCL8, CCL3, IFI16, and IRF1. These highly-expressed genes were significantly enriched in numerous biological pathways and functional terms associated with autoimmune diseases (FDR < 0.05). Conclusions and relevanceTo the best of our knowledge, this is the first study to integrate genetic information with single cell sequencing data for parsing genetics-influenced liver cells for PBC risk. We identified that ORMDL3- cholangiocytes play important immune-modulatory roles in the etiology of PBC. Key pointsO_ST_ABSQuestionC_ST_ABSAre genetics factors influenced liver cell subpopulations and its immune microenvironment for PBC? FindingsIn this comprehensive genomics study based on multi-omics data, genetic determinants were significantly enriched in cholangiocytes and immune cells including subsets of macrophage, dendritic cells, and T cells. ORMDL3- cholangiocytes have crucial immune-modulatory roles in developing PBC. MeaningFindings suggest that integration of single cell sequencing data with GWAS summary statistics contribute to pinpoint PBC-relevant cell types and risk genes.

Since diagnostic specimens retrieved from pathology cannot be altered (except for sectioning) for ethicolegal reasons, we describe a method that combines multiple patient specimens, cut from individual blocks, allowing the measurement of several tissues in one capture array. This results in a cost-effective means to increase sample size to permit statistically robust data outputs and to fast-track clinical implementation. Importantly, our tissue multiplexing method is compatible with the standard 10X Genomics supported protocols. We have used skin biopsies in this instance as this is our primary area of research, but this method can be extended to any tissue of interest.

Homologous recombination deficiency (HRD) testing has been approved by FDA for selecting epithelial ovarian cancer (EOC) patients who may benefit from the first-line poly (ADP-ribose) polymerase inhibitor (PARPi) maintenance therapy. However, the effects of HRD on the clinical outcomes of first-line chemotherapy and first-line PARPi maintenance therapy have not been rigorously evaluated in Chinese EOC patients. Here, we developed an HRD assay and applied it to two large Chinese EOC patient cohorts. In the first-line adjuvant chemotherapy cohort (FACT, N = 380), HRD status significantly improved PFS (median, 15.6 months vs. 9.4 months; HR, 0.688; 95% CI, 0.526 to 0.899; P = 0.003) and OS (median, 89.5 months vs. 60.9 months; HR, 0.636; 95% CI, 0.423 to 0.955; P = 0.008). In the first-line PARPi maintenance therapy cohort (FPMT, N = 83), HRD status significantly improved PFS (median, NA vs 12 months; HR, 0.438; 95% CI, 0.201 to 0.957; P = 0.033) and OS (median, NA vs NA months; HR, 0.12; 95% CI, 0.029 to 0.505; P = 0.001). Our results demonstrate that HRD status is a significant predictor for PFS and OS in both first-line chemotherapy and first-line PARPi maintenance therapy, providing strong real-world evidence for conducting genetic testing and improving clinical recommendations for Chinese EOC patients.

Previous work demonstrated that ectodermal cells exhibit greater susceptibility to SARS-CoV-2 infection than endodermal and mesodermal cells, raising concerns about potential vulnerability of the developing nervous system. We hypothesized that neural stem cells (NSCs) derived from human ectoderm are susceptible to SARS-CoV-2 infection. Using pseudotyped viral particles representing both wild-type and Omicron spike variants, we confirmed efficient infection of NSCs, with Omicron variants preferentially utilizing endocytosis-mediated entry. Inhibition of endocytosis with filipin and OcTMAB significantly reduced infection across all spike variants. Low glycosylation levels on NSCs facilitated viral entry, and enzymatic removal of glycosylation increased their susceptibility. Ectodermal infection by SARS-CoV-2 raises serious concern for potential teratogenic effects on the nervous system, possibly causing latent or subclinical anomalies not immediately evident at birth. Therefore, future clinical studies and long-term surveillance of infants and children exposed in utero are necessary to investigate and identify potential neurodevelopmental deficits.

Transplanting human stem cell-derived islets (SC-islets) is a promising therapy for insulin-dependent diabetes. While functional SC-islets have been produced for clinical application, immune rejection by the host remains a challenge. Present attempts, including chronic immunosuppression and/or physical encapsulation, have some disadvantages. Here we explore a strategy to induce an immune-tolerant environment based on the immune privilege observed in the male gonad. Sperm appears after the maturation of the immune system and development of systemic self-tolerance and the testis protects these autoreactive germ cells by the physical structure of blood-testis-barrier (BTB) and active local immunosuppression. Human SC-islets transplanted into the mouse testis can be physically protected by the BTB and we find that the testis secretes cytokines that induce a population of regulatory T cells (Tregs) that express both CD4 and CD8. We identified cytokines secreted by testis and used a cocktail of IL-2, IL-10, and TGF-{beta} for in vitro co-culture and in vivo transplantation demonstrating improved survival of SC-islets and the induction of Tregs. One Sentence SummaryInducing local immunotolerance by suppressive cytokines for islet Transplantation.

This study investigated the human papillomavirus (HPV) infection status, genotype distribution, and associated risk factors among women in Suzhou to provide a theoretical basis for controlling cervical cancer and guiding vaccine development. From November 2022 to April 2023, 566 qualified participants undergoing health examinations at Suzhou Municipal Hospital were enrolled. Cervical exfoliated cells were tested for 21 HPV genotypes using fluorescence quantitative PCR, and data on general information, lifestyle, and clinical history were collected via questionnaire. The overall HPV infection rate was 11.31% (64/566). The most prevalent genotypes were HPV16, HPV52, and HPV58, each with an infection rate of 1.77%. Single infections predominated (8.83%, 50/566), with HPV16 (18%), HPV58 (14%), HPV51 (12%), and HPV81 (12%) being the most common. Multiple infections accounted for 2.47%, primarily dual infections (78.57%). Infection rates varied significantly by age, with the highest prevalence in women aged [&ge;]50 years (24.37%), followed by those aged 40-49 (8.42%) and [&le;]39 (7.35%). Significant risk factors for HPV infection included older age (OR=1.075, P<0.01), secondhand smoke exposure (OR=2.126, P<0.05), a history of underlying diseases, and a history of cervical surgery. In conclusion, the HPV infection rate in Suzhou remains considerable, with genotypes 16, 52, and 58 being predominant and single infections most common. Older age, secondhand smoke exposure, underlying diseases, and cervical surgery history are key risk factors, highlighting the need for targeted prevention and comprehensive control strategies.

Our previous studies have indicated that oxidative stress and inflammation can dose-dependently induce increased skins green autofluorescence (AF) of mice, which results at least partially from keratin 1 (K1) cleavage. Increased green AF was also found in patients skin of several major diseases, which may become a novel biomarker for non-invasive diagnosis. We also found age-dependent changes of the skins green AF of natural populations. In this study we tested our hypothesis that there are growth stage-dependent changes of K1 and keratin 10 (K10) levels in the skin of mice, which may underlie the age-dependent changes of the skins green AF. We found that in the skin of both mices back and ears, there were growth stage-dependent changes of the levels of K1 and K10 as well as the basal green AF. The K1 and K10 levels in the backs skin were significantly different from those in the ears skin. There were also growth stage-dependent changes of the UVC-induced changes of K1 and K10 levels of both the ears and the back. Collectively, our study has provided first evidence showing growth stage-dependent and differential changes of the levels of K1 and K10 as well as skins green AF in the back and the ears of mice under basal conditions and after UVC irradiation. These findings are valuable for understanding the age-dependent changes of the skins green AF of natural populations, which are also important for establishing the keratins AF-based method for non-invasive diagnosis of diseases.

BackgroundOral squamous cell carcinoma of the gingivo-buccal region (OSCC-GB) has the highest incidence among males and the second highest overall among all cancers in India, emphasizing the need for precise molecular classifications to guide personalized therapy. MethodsWe performed bulk RNA sequencing on tumor and adjacent normal tissue samples from 72 OSCC-GB patients, as well as leukoplakia tissue from 25 patients with concurrent leukoplakia. FindingsOur analysis revealed activated epithelial-mesenchymal transition, angiogenesis, and cell-cycle function in OSCC-GB. We found significant enhancement of glycolysis and reduction in oxidative phosphorylation, which are hallmarks of the Warburg effect. Immune profiling indicated enriched immune-related genes and cells in tumor tissues. We identified two distinct patient subtypes, one of which exhibited higher immune cell infiltration and showed potential for greater responsiveness to immune checkpoint inhibitors. CD226, CD38, and KBTBD8 were identified as potential biomarkers for classifying OSCC-GB patients and were validated in an independent cohort. Significantly more M1 macrophages and CD4+ T-cells in leukoplakia tissue than the normal indicate activated host defense mechanisms in pre-malignant lesions, highlighting the potential for early intervention to prevent malignancy. TCGA-HNSC data exhibited similar gene set enrichments, including glycolysis and immune-related pathways. However, unique profiles in a subset of TCGA-HNSC patients highlight the molecular heterogeneity of head and neck cancers. ConclusionOur findings underscore the critical role of understanding these pathways in cancer biology and immunology, essential for developing effective treatment strategies for oral cancer and immunotherapy.