Fine-needle aspiration (FNA) is the gold-standard diagnostic procedure for the initial evaluation of thyroid nodules because it is minimally invasive, cost-effective, and highly accurate for distinguishing benign from malignant lesions. However, conventional smear preparation is frequently affected by operator-dependent variability, uneven cell distribution, blood contamination, and air-drying artifacts, all of which may compromise specimen adequacy and diagnostic performance. Liquid-based cytology (LBC) has emerged as an effective alternative, offering standardized specimen processing, improved cellular preservation, and enhanced slide quality. Among the available LBC platforms, the ThinPrep system is one of the most extensively validated technologies for thyroid cytopathology and is increasingly integrated into routine clinical practice and molecular diagnostic workflows. This protocol provides a comprehensive, standardized workflow for processing thyroid FNA specimens using the ThinPrep liquid-based cytology system. The protocol describes specimen collection, transportation, fixation, processing with the ThinPrep 5000 processor, slide preparation, staining, and quality control procedures to ensure optimal cytomorphological evaluation. Recommendations for acceptable specimen preservation, including transport in PreservCyt solution or alternative fixation methods, are also provided. In addition, the protocol includes procedures for recovering the residual cellular material preserved in the ThinPrep vial for paraffin block processing, which allows for complementary studies, e.g., immunohistochemistry, fluorescence in situ hybridization (FISH), DNA and RNA extraction, and next-generation sequencing (NGS), thus expanding the diagnostic utility of thyroid cytology beyond conventional microscopic examination. When properly executed, this protocol generates high-quality monolayer cytology preparations suitable for standardized interpretation according to The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) while preserving residual material for downstream molecular analyses. Compared with conventional smear cytology, the ThinPrep system improves specimen reproducibility, reduces background artifacts and inadequate samples, facilitates laboratory standardization, and supports precision medicine applications. This protocol provides a practical and reproducible approach for both clinical diagnostic laboratories and translational research. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Simultaneous preparation of ThinPrep cytology slides and cell blocks from thyroid fine-needle aspiration specimens using liquid-based cytology Alternate Protocol: Alternative ThinPrep FNA processing using CytoLyt and PreservCyt.

This study successfully synthesized UiO-66/ZnO composites via a hydrothermal method, and their structure and morphology were comprehensively characterized. Using tetracycline (TC) as a model pollutant, the sonocatalytic performance of the composite was systematically evaluated. The optimal degradation conditions were determined as follows: UiO-66 mass ratio of 30% in composite, catalyst dosage of 1 g/L, initial TC concentration of 50 mg/L, ultrasonic power of 500 W, and reaction time of 60 min, achieving a removal efficiency of 95.61 ± 0.82%. in addition, possible TC degradation pathways were identified using LC-MS analysis. The results of evaluating the antibacterial properties of this material showed that the antibacterial rate of 25 μg/mL UZ-30 against Escherichia coli (E. coli) was only 9.69 ± 3.13%. When combined with ultrasound, the antibacterial rate against E. coli could reach 98.54 ± 0.51%. The outstanding performance of the UiO-66/ZnO composite was attributed to the formation of a type-II heterojunction, which effectively separated the photothermal-generated charge carriers produced under ultrasonic excitation, thereby generating more active species. This not only led to the effective degradation of antibiotics but also enabled efficient elimination of bacteria. The UiO-66/ZnO composite material prepared in this study possessed dual functions of degrading antibiotic pollution to prevent the emergence of drug-resistant bacteria and inhibiting pathogenic microorganisms, which had broad application prospects in the field of combined water purification treatment and disinfection.

Pulmonary arterial hypertension (PAH) is characterized by a neurohumoral imbalance. We studied the effect of PAH on transcriptional activity in pulmonary artery preparations. In rats with PAH induced by monocrotaline (60 mg/kg), pulmonary artery preparations were isolated and whole-transcriptome RNA sequencing was performed. The enrichment score of gene collections from the Gene Ontology and KEGG databases was determined. Significant gene sets were grouped into clusters, and genes exhibiting the greatest changes in expression and representation within these clusters were identified. The representation of functional domains in protein sequences was characterized for the key genes. PAH led to a significant increase in the enrichment score of pathways associated with contractile elements and the cytoskeleton of muscle cells (including cardiomyocytes) and a decrease in the score for pathways associated with the synthesis of ribosomal components. Clustering of the pathways revealed changes in gene expression indicative of impaired thyroid signaling.

Bone marrow mesenchymal stem cells (BMSCs) are stromal cells that can self-renew. Exosomes (EXOs) perform many biological functions particularly in cell-cell communication. Peripheral blood mononuclear cells (PBMNCs) have the ability to differentiate into numerous mature functional cell types in specific microenvironments. This study aimed to compare the possible ameliorating effect of BMSCs, EXOs, and PBMNCs on hypothyroidism-induced testicular damage in adult male rats. Forty‑eight adult male albino rats were divided randomly into five groups: control, hypothyroidism (HT), BMSCs-treated, EXOs-treated, and PBMNCs-treated groups. Testes and thyroids were dissected and prepared for biochemical, histological, and immunohistochemical studies. After induction of HT, the results showed an apparent decrease in testes weight and abnormal semen analysis. Additionally, a significant decrease in antioxidant markers, thickness of the germinal epithelium, PAS staining, and testes BCL2 anti-apoptotic marker. It showed histological changes. Additionally, a significant decrease in SIRT-1 and ZO-1immune stained cells. A significant increase in testes BAX apoptotic marker, FOXO-1 immuno-expression, and deficient blood testes barrier with abnormal spermatogenic cells in electron microscopy were also detected. Improvement in all mentioned parameters in treated groups was noticed. Our study suggested that PBMNCs holds promise for alleviating hypothyroidism in rats on biochemical, histological, and immunohistochemical basis.

Hypothyroidism is a chronic thyroid disorder associated with fatigue, cold intolerance, weight gain, constipation, menstrual disturbance, hair fall, mood changes, cognitive slowing, and reduced quality of life. This narrative review synthesises published literature on homoeopathic management of hypothyroidism, with emphasis on individualised prescribing, reported symptom and biochemical outcomes, quality of life, and methodological and reporting limitations. The final review incorporated 53 clinical, methodological, and contextual sources. The literature includes observational studies, case series, case reports, and other descriptive sources involving overt hypothyroidism, subclinical hypothyroidism, and autoimmune thyroiditis. Reported approaches include individualised remedies, Thyroidinum and other sarcodes, intercurrent prescriptions, and follow-up-based modification of treatment. Reported outcome domains include thyroid-stimulating hormone, free triiodothyronine, free thyroxine, thyroid antibodies, fatigue, bowel function, menstrual symptoms, weight change, hair fall, mood, sleep, and quality of life. The available evidence is heterogeneous and predominantly uncontrolled, with variation in thyroid subtype, intervention reporting, potency, repetition, concurrent treatment, follow-up duration, and outcome assessment. Reported improvements therefore cannot be assumed to represent treatment-specific effects and may be influenced by natural disease variation, spontaneous thyroid-stimulating hormone fluctuation, regression to the mean, placebo or contextual effects, and concurrent care. A scientifically established mechanism for specific clinical effects of highly or ultra-highly diluted homoeopathic preparations has not been demonstrated, and the broader evidence base does not establish consistent treatment-specific efficacy beyond placebo or contextual effects. Homoeopathic care should not replace or delay indicated thyroid hormone therapy, particularly in overt hypothyroidism, pregnancy, severe thyroid dysfunction, or clinical deterioration. More rigorous controlled studies with clearly defined thyroid subtypes, standardised clinical and biochemical outcomes, documentation of concurrent conventional treatment, and adequate follow-up are required to determine whether any specific therapeutic effect exists.

Amphotericin B (AmB) remains the gold-standard antifungal agent, yet its clinical utility is severely limited by dose-dependent hemolytic and nephrotoxic side effects arising from its aggregation-prone nature and non-selective membrane binding. To address this challenge, we developed a polymer prodrug by conjugating AmB to oxidized chondroitin sulfate (CSox) via a Schiff-base linkage designed to be pH-labile, selected for CS's CD44-targeting capability, intrinsic anti-inflammatory activity, and favorable polyelectrolyte properties. The resulting CSox-AmB conjugate self-assembled into uniform micelles (141.60 ± 4.03 nm, zeta potential -25.47 ± 0.59 mV) with a drug loading of 7.07 ± 0.13%, maintaining AmB predominantly in its monomeric state (A348/A409 = 0.865 < 1). The prodrug exhibited sustained release (∼68% at 24 h) while preserving antifungal activity comparable to free AmB against Candida albicans. Concurrently, it attenuated mammalian cell cytotoxicity by 2.3- to 2.5-fold and markedly attenuated hemolytic activity in vitro (<1% at 25 μg·mL[-1]). This integrated design-conjugation enabling self-assembly, assembly enabling monomeric stabilization, and controlled release enabling efficacy with markedly reduced toxicity-positions CSox-AmB as a promising prodrug strategy that uncouples the antifungal activity of AmB from its inherent toxicities, warranting further in vivo evaluation toward clinical translation.

Background/Objectives: Reliable molecular testing is essential for selecting targeted therapies for thyroid tumors; however, assay performance can be affected by pre-analytical tissue conditions. This retrospective, single-center study evaluated real-world formalin-fixed, paraffin-embedded (FFPE) specimens to identify pathology practices associated with successful companion diagnostic testing using the next-generation sequencing (NGS)-based Oncomine Dx assay and the polymerase chain reaction (PCR)-based BRAF3 assay widely used in routine clinical care in Japan. Methods: We retrospectively analyzed 177 cases with thyroid tumors tested at a high-volume thyroid center in Japan between 2022 and 2026. Results: All 79 specimens analyzed using the BRAF3 assay yielded successful test results, including specimens fixed in unbuffered formalin and archival blocks stored for up to 27 years. In contrast, the Oncomine Dx assay was successful in 93 of 98 specimens (94.9%), with all five failures occurring in specimens fixed in unbuffered formalin. DNA failure was more common than RNA failure. All five Oncomine Dx failures occurred in archived tissue blocks stored for 3-9 years, all of which had been fixed in unbuffered formalin. The BRAF3 test had a shorter turnaround time and was more frequently used in rapidly progressive tumors, such as anaplastic thyroid carcinoma. Conclusions: The NGS-based Oncomine Dx assay showed reduced success in samples stored long-term or in unbuffered specimens, whereas RNA preservation was high. In contrast, the PCR-based BRAF3 assay remained highly reliable even in long-stored archival blocks. Because fixative type and storage duration were confounded in this cohort, the independent effects of these variables could not be determined. The study's findings provide practical real-world insights into how specimen handling, fixation, and storage conditions may influence the reliability of genomic diagnostics in routine practice.

Thyroid nodules are a prevalent endocrine disease with rising global incidence. Modern clinical management relies on standardized workflows stratified by nodule size, TI-RADS ultrasound risk grading, fine-needle aspiration biopsy (FNA) indications, cytological results, thyroid function and malignant risk, including active surveillance, thermal ablation and surgical resection as definitive interventions. While western medicine achieves precise diagnosis and radical treatment, it still faces limitations such as recurrent benign nodules, postoperative complications and long-term hormone side effects. Traditional Chinese Medicine (TCM), as a complementary and adjuvant therapy rather than a replacement for definitive western treatment, has accumulated mature clinical experience for auxiliary intervention of thyroid nodules. This review firstly systematically sorts out modern diagnostic techniques, epidemiological characteristics, genetic susceptibility and core pathophysiological mechanisms of thyroid nodules, and fully summarizes guideline-based standardized western medical treatment paradigms. Then we systematically expound the TCM etiology, pathogenesis, syndrome differentiation theory, classical compound preparations (Xiaojin Capsule, Neixiao Tablet, external Jie Jian gel), single herbal medicines and isolated active monomers (diosgenin), and clarify their respective applicable scenarios including preoperative conditioning, perioperative support, postoperative rehabilitation and symptomatic relief after ablation. We further stratify all cited research evidence by study type (preclinical experiments, observational trials, randomized controlled trials and meta-analyses) to avoid overstating clinical efficacy based solely on animal and cell experiments. Meanwhile, we summarize integrated Chinese-Western combined regimens and current research advances in TCM mechanism research. Finally, we independently discuss the controversies, safety risks, standardization defects and clinical research limitations of TCM intervention, and put forward future research directions. This review aims to clarify the auxiliary positioning of TCM in the overall thyroid nodule treatment system, provide comprehensive integrated Chinese-Western clinical reference, and promote standardized and translational research of TCM for thyroid nodules.