The aim of this study was to elucidate the risk of developing voice disorders among peri- and post-menopausal female hormone replacement therapy (HRT) users.

A retrospective cohort study was conducted using the TriNetX Global Collaborative Network. Females aged 40-60 years old were included and stratified into two cohorts: HRT users (n = 16,586) and HRT non-users (n = 248,725) while excluding for head and neck radiation/neoplasms, smoking, benign laryngeal lesions, thyroid disorders, gender dysphoria, and any other systemic hormone use. Voice and resonance disorders (VRD) and dysphonia were separately assessed at 3-month intervals post-HRT initiation after propensity score-matching for age, sex, race, and ethnicity. Odds ratios (ORs) with 95% confidence intervals (CIs) and risk differences (RD) were generated to compare outcomes.

HRT users had significantly higher odds of dysphonia, but not VRD, within 0-9 months (OR 1.72; 95% CI (1.01-2.95), OR 1.66; 95% CI (0.99-2.79), respectively). For HRT users with elevated BMI, the 1-year VRD incidence was not significantly different than non-HRT users (RD 0.19%, p = 0.069). Overall, the incidence of VRD and dysphonia within 1 year in both groups was < 0.5% and showed no significant difference at most times between groups.

HRT may not have any significant clinical impact on the peri- and post-menopausal voice. Although higher odds were noted earlier after treatment, HRT users did not have higher odds of voice issues after 1 year. This study highlights the lack of consensus in literature and urges future research to fully understand the impact of HRT on peri- and post-menopausal voice.

Cancer is the most threatening disease worldwide. Noticeably, most cancer patients are immune-compromised, so they are more susceptible to infections. The combination of the tyrosine kinase inhibitor nilotinib (NIL) with broad-spectrum antibiotics such as the fluoroquinolone ofloxacin (OFL) is clinically rationalized based on NIL's immunosuppressive effects and the need for infection prophylaxis in cancer patients. However, there is no published method for their in-vivo simultaneous determination in clinical biological specimens to support their pharmacokinetics and therapeutic monitoring. This study focuses on the development of a miniaturized capillary electrophoresis system-assisted with photodiode array detection (CE-PDA) for the simultaneous determination of NIL and OFL in clinical settings for analysis of plasma samples containing NIL and OFL. The conditions of electrophoretic separation of NIL, OFL, and sarafloxacin (SAR, as an internal standard), and the analytical procedures were successfully established. The electrophoretic separation of NIL, OFL, and SAR, as the internal standard, was achieved utilizing a fused silica capillary (58 cm length × 75 μm internal diameter) maintained at 25 °C using a background electrolyte solution composed of borate buffer (20 mM, pH 9.3) at a separation voltage of 30 kV. The linear range of the CE-PDA method in spiked rat plasma was determined to be 0.5-80 µg mL^- 1 for NIL and OFL. The CE-PDA approach exhibited maximum sensitivity, with lower limits of quantitation (LLOQ) of 0.3 µg mL^- 1 for NIL and 0.4 µg mL^- 1 for OFL. These LLOQs were experimentally validated according to US-FDA guidlines and were lower than the calibration range's starting point to ensure reliable quantification. In addition, the CE-PDA system was validated and subsequently applied to real plasma samples withdrawn from rats received concurrent administration of NIL and OFL. The plasma protein was precipitated by acetonitrile. The validation parameters were determined and found to comply with the requirements for the Food and Drug Administration (FDA) guidelines for validation of bioanalytical procedures. The sustainable features of the system and its environmental impact were confirmed using seven different metric tools. Finally, the proposed CE-PDA is time-saving and cost-effective, enabling its beneficial applications in the therapeutic drug monitoring of NIL and OFL in oncology clinics.

The clinical behavior and molecular underpinnings of rare cases of poorly differentiated thyroid carcinoma (PDTC) diagnosed in young adults are not established. We evaluated 13 cases of PDTC in adult patients ≤ 45 years of age. The median age at diagnosis was 40 years (range 27 - 45 years). The median tumor size was 3.7 cm. The median mitotic count was 6 per 2 mm². Necrosis was present in 7 (54%) cases. Eleven (85%) cases were either encapsulated with extensive (≥ 4 foci) angioinvasion or widely invasive. Molecular analysis was available for nine (69%) cases and revealed two (22%) with PAX8::PPARG fusions, two (22%) with NRAS alterations, one (11%) with a DICER1 variant, one (11%) with alterations in ATM, CDKN1A and TERT, one (11%) with a TSC2 deletion and TERT copy number gain, and two (22%) with variants of uncertain significance only. Of the 12 (92%) patients with follow-up data available (median 6.2 years, range 1.5-16.4 years), the only patient who died of disease had distant metastases at diagnosis. Of the remaining patients with initial M0 disease, only one had disease recurrence, developing local recurrence and distant metastasis with persistent disease at last follow up. Our findings suggest that PDTC in adults ≤ 45 years of age at diagnosis, may have a better prognosis than is typically associated with PDTC despite high-grade features and invasive growth. Additionally, we found a higher rate of PAX8::PPARG fusions and a lower rate of TERT alterations, suggesting that the molecular profile might explain the improved survival.

There is a lack of prospective clinical trials on the therapy of salivary gland cancer (SGC) due the rarity and large variety of histological subtypes. This study aimed to compare overall survival (OS) after primary surgery of SGC followed by postoperative radiotherapy (PORT) versus radiochemotherapy (PORCT) in curative intent. A systematic review and meta-analysis were performed on studies reporting OS after the two postoperative therapy modalities using hazard ratios (HRs) or allowing to calculate the HRs from the data. These studies were identified from PubMed, Web of Science, and Cochrane Library databases until October 2024. Pooled HR with 95% confidence interval (CI) is reported with random-effects or fixed-effects models. The search yielded 1,074 publications, of which 11 retrospective clinical studies with 26,612 adult patients could be included. None of the studies found a statistically significant difference between PORT and PORCT. Likewise, this meta-analysis revealed no statistically significant difference between PORT and PORCT. The pooled HR based on a common effect model was 1.065 (CI: 0.998-1.137) with low between study heterogeneity (I²=24.1%) and heterogeneity variance τ² = 0.009. A meta-regression only revealed one factor with influence on the HR. The proportion of T4 patients in the total study shows a HR = 1.153 for a 10% change 95% CI: 1.014,1.314). This meta-analysis, based on a highly confounded observational setting, does not permit a reliable conclusion as to whether PORCT leads to better OS than PORT as adjuvant therapy for patients with SGC. PORCT might be beneficial for high-risk subgroups. However, this was not possible to determine in this meta-analysis as all included studies were retrospective and heterogeneous leading to a high degree of uncertainty about the therapy effects. More robust data from high-quality cohort studies or randomized controlled studies are needed. There is an urgent need for alternative and more effective multimodal treatment concepts for SGC.

Pancreatic cancer has poor prognosis, and immunotherapy efficacy is limited by PD-L1 stability via GFAT1-mediated glycosylation. We developed LNP@2DG-DON, a liposomal nanoparticle co-delivering glycolysis inhibitor 2DG and GFAT1 inhibitor DON, to enhance antitumor immunity and metabolic targeting.

LNP@2DG-DON nanoparticles were synthesized using the thin-film dispersion method and characterized for size, stability, drug loading, and release profiles. In vitro studies assessed cytotoxicity, apoptosis, migration, and invasion in ASPC-1 and PANC-1 pancreatic cancer cell lines. In vivo efficacy was evaluated in a subcutaneous xenograft mouse model, measuring tumor growth, immune cell infiltration, and survival. Western blotting, flow cytometry, and immunohistochemistry were employed to analyze molecular and cellular mechanisms.

LNP@2DG-DON showed uniform size (100-120 nm), high drug loading (6.84-9.27%), and sustained release. It significantly reduced cell viability, induced apoptosis, and inhibited metastasis in vitro. In vivo, it suppressed tumor growth, prolonged survival, and increased CD8+/NK cell infiltration while reducing immunosuppressive cells. Mechanistically, it downregulated PD-L1/GFAT1 and activated pro-apoptotic pathways.

The dual-targeting LNP@2DG-DON nanoparticle synergistically combines metabolic and immune modulation, demonstrating superior antitumor effects compared to single-agent therapies. This approach represents a promising strategy for pancreatic cancer treatment, warranting further clinical investigation.

Breast cancer (BC) is the most prevalent cancer among women, and retrieving the anti-tumor function of the immune system seems a promising treatment approach for its crucial role in combating cancer cells. In this study, we evaluated the impact of a combination therapy containing a TAK1 inhibitor, Takinib (Tak), a metabolic regulator, Metformin (Met), and an immunostimulant, Lentinula edodes mycelia extract (LEME) on enhancing the immune system's anti-tumor activity in BALB/c mice bearing triple-negative breast cancer (TNBC).

BALB/c mice were used to induce TNBC tumors and evaluate tumor growth inhibition. The number of CD8⁺ CD28⁺ T cells was determined by immunofluorescence assay, the expression of MUC1 protein was assessed by Western blot, while the expression of TOX, NR4A1, and TIM-3 genes was evaluated by real-time PCR in mouse-derived tumor tissues. MTT assay was performed on different BC cell lines to assess cell viability.

Molecular docking results revealed significant interactions between Tak and Met with TOX and NR4A1. The combination treatments of Tak, Met, and LEME significantly decreased tumor volume/weight in mice and also significantly increased the number of infiltrated CD8⁺ CD28⁺ T cells, reduced MUC-1 protein expression, and decreased the expression of TOX, NR4A1, and TIM-3 genes in mouse tumor tissue. Tak, Met, and their combination significantly decreased the cell viability of different BC cell lines.

This study suggests that the Tak-Met-LEME combination treatments may inhibit BC progression by increasing CD8⁺ CD28⁺ T cell population in tumor tissue and decreasing tumor progression.

Osteosarcoma (OS) is a highly aggressive primary bone malignancy with a strong predilection for lung metastasis, and the formation of a lung pre-metastatic niche represents a key early event driving OS distant dissemination. Elucidating the molecular mechanisms of pre-metastatic niche establishment is therefore critical for developing novel anti-metastatic therapies for OS. However, the mechanisms by which OS cells initiate the formation of a lung pre-metastatic niche remain poorly understood. Exosomes, as key mediators of intercellular communication, play a pivotal role in promoting tumor metastasis by shuttling functional biomolecules to remodel the microenvironment of distant target organs. In this study, we found that MG63 OS cells secreted exosomal miR-27a-3p, which targeted TNFAIP3 in lung fibroblasts, resulting in decreased TNFAIP3 protein expression. This downregulation enhanced the metastatic potential of OS cells toward lung fibroblasts. TNFAIP3 was identified as a direct target of miR-27a-3p, and TNFAIP3-Mut abrogated the regulatory effect of miR-27a-3p on TNFAIP3 expression, confirming the specific targeting relationship. Mechanistically, miR-27a-3p-mediated suppression of TNFAIP3 abrogated its negative regulation of the NF-κB pathway, leading to increased transcription of downstream inflammatory cytokines IL-1β, IL-6, and IL-8. The resulting pro-inflammatory microenvironment contributed to the establishment of a lung pre-metastatic niche, thereby promoting OS lung metastasis. These findings underscore the critical role of OS-derived exosomes in lung metastasis and suggest that exosomal miR-27a-3p may serve as a promising therapeutic target in OS lung metastasis.

Epidermal growth factor receptor (EGFR) "membrane-cytoplasmic-nuclear translocation" occurs in EGFR-19del lung adenocarcinoma (LUAD) following resistance to tyrosine kinase inhibitors (TKIs). This study aimed to elucidate the mechanism of TKI resistance conferred by nuclear EGFR-19del.

RNA sequencing and immunohistochemistry were performed to assess histone deacetylase 7 (HDAC7) expression in LUAD with TKI resistance. Functional assays were performed both in vitro and in vivo to assess the effects of changes in HDAC7 expression on the malignant phenotype of LUAD cells and drug sensitivity to TKIs. Mass spectrometry and dual-luciferase assays were performed to verify the effect of changes in HDAC7 expression on the Hippo pathway. Chromatin immunoprecipitation and coimmunoprecipitation assays were conducted to clarify the potential role of EGFR-19del in the cell nucleus.

EGFR-19del nuclear translocation correlated with elevated HDAC7 expression in TKI-resistant cells. HDAC7 overexpression promoted malignancy and reduced TKI sensitivity, whereas HDAC7 knockdown or TSA treatment suppressed tumour growth and enhanced TKI sensitivity in vivo. Mechanistically, HDAC7 interacts with large tumour suppressor kinase 1 (LATS1) and promotes its deacetylation at K688, which reduced T1079 phosphorylation, thereby inhibiting the Hippo pathway. Concurrently, nuclear EGFR-19del acts as a coactivator to accelerate HDAC7 transcription through signal transducer and activator of transcription 3 (STAT3).

We elucidated the underlying mechanism by which nuclear EGFR-19del inhibits the Hippo pathway; these results indicate that TKIs and HDAC inhibitors may serve as a potential therapeutic strategy to reduce drug resistance in LUAD with EGFR-19del.

Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease for which novel therapeutic approaches are urgently needed. Transforming Growth Factor-β (TGF-β) plays a central role in IPF pathogenesis by activating lung fibroblasts. Inhibitor of DNA binding (ID) proteins are regulated by TGF-β; however, their role in IPF remains poorly understood. We aimed to evaluate the regulation of ID proteins in IPF and to determine their functional role in human lung fibroblasts (HLF) in vitro and pulmonary fibrosis in vivo.

ID protein expression was assessed in lungs and lung fibroblasts from mice and patients with pulmonary fibrosis. In vitro, the effects of ID1/ID3 inhibition and overexpression on HLF proliferation, migration and differentiation into myofibroblasts were evaluated. Genetic and pharmacological approaches were used in vivo to determine the role of ID1/ID3 in pulmonary fibrosis.

ID1/ID3 levels were elevated in lungs and lung fibroblasts from mice and patients with pulmonary fibrosis, as well as in HLFs treated with TGF-β. ID1/ID3 knockdown reduced proliferation, migration and differentiation of healthy and IPF-derived HLFs. Bleomycin-exposed ID1/ID3 double KO mice exhibited improved lung function and reduced fibrosis compared with WT mice. Pharmacological inhibition of ID1/ID3 decreased HLF proliferation, migration and differentiation in vitro and attenuated pulmonary fibrosis in vivo. Lung-specific inhibition of ID1/ID3 using adeno-associated viruses expressing short hairpins targeting ID1 and ID3 also reversed pulmonary fibrosis in mice. Mechanistically, ID1/ID3 inhibition reduced fibroblast proliferation through regulation of cell cycle genes and attenuated fibroblast differentiation via the MEK/ERK pathway.

Simultaneous inhibition of ID1 and ID3 attenuates pulmonary fibrosis. Targeting ID1/ID3 represents a potential novel therapeutic strategy for IPF.

Renal cell carcinomas (RCC) are resistant to chemotherapy and radiotherapy, and effective treatment options remain limited. Immunotherapy has emerged as a promising approach, and circular RNAs (circRNAs) are increasingly recognized as key regulators of tumor immunity. This study aims to elucidate the regulatory relationship between circPVT1 and antitumor immunosuppression in RCC.

CircPVT1 expression and its prognostic value were analyzed in RCC samples. The effects of circPVT1 knockdown on RCC cell proliferation, invasion, and metastasis were examined in vitro. Singlecell sequencing was used to assess macrophage infiltration in circPVT1high versus circPVT1low groups. ELISA was performed to measure secretion of IL4, IL10, and TGFβ by tumor cells. In addition, nanotherapeutic system delivering circPVT1 inhibitors was tested for its antitumor efficacy.

CircPVT1 was highly expressed in RCC and correlated with poor prognosis. Knockdown of circPVT1 significantly suppressed cell proliferation, invasion, and metastasis. Singlecell sequencing revealed increased macrophage infiltration in the circPVT1high group. ELISA results demonstrated that circPVT1 knockdown reduced tumor cell secretion of IL4, IL10, and TGFβ, thereby promoting macrophage polarization toward the M1 phenotype. Mechanistically, circPVT1 promoted RCC progression by regulating EMT. Furthermore, a nanotherapeutic system containing circPVT1 inhibitors effectively inhibited RCC growth.

These findings indicate that circPVT1 plays a critical role in RCC immune evasion by modulating macrophage polarization and EMT. Therefore, circPVT1 may be a predictor of ccRCC immune evasion and a potential therapeutic target.