The spectrum of hyperglycemia and its association with overall survival (OS) in patients receiving immune checkpoint inhibitors (ICIs) are not characterized. We retrospectively analyzed 4,119 patients who had received ≥ 1 dose of PD-1 or PD-L1 inhibitors during a period from July 1st 2018 to March 1st 2022 at the First Affiliated Hospital with Nanjing Medical University. We assessed the characteristics of patients with hyperglycemic during ICIs. Multivariable Cox regression models and Propensity score matching (PSM) were utilized to analyze the risk of hyperglycemic and OS differences. After excluded 1,283 patients without follow-up fasting blood glucose (FBG) data, 2,836 patients were included. 24.33

Background: Immune Checkpoint Inhibitor (ICPi) therapy has revolutionized cancer treatment but can lead to immune-related adverse events (irAE), including thyroid dysfunction. The impact of ICPi on patients with pre-existing autoimmune thyroid diseases (PATD), particularly the development of Graves' disease, remains poorly understood. Case Description: We provide the first complete case of Graves' disease with ICPi therapy in a patient who already had Hashimoto's thyroiditis.. The patient, a 52-year-old male, was diagnosed with lung adenocarcinoma and received Atezolizumab. Clinical evaluation revealed hyperthyroidism, confirmed by elevated thyroid hormones and autoantibodies (TRAb and TSAb). The patient was managed with methimazole and demonstrated a transient hyperthyroid phase followed by persistent hypothyroidism. Only 16 confirmed cases of Graves' disease induced by ICPi were reported. We conducted a review to investigate the clinical characteristics, risk factors, and prognosis trends associated with ICPi-induced Graves disease in PTAD patients. Additionally, changes in thyroid function and autoantibodies during and after ICPi treatment are examined. Conclusion: This case underscores the importance of monitoring thyroid function and autoantibodies in patients with PATD undergoing ICPi therapy. The findings suggest distinct differences in the humoral immune response between ICPi-induced and spontaneous Graves' disease, necessitating further research into autoantibody dynamics and their relationship with cellular immunity in these patients.

ABSTRACT Alveolar macrophages (AMs) serves as a frontline innate barrier against pulmonary bacterial invasion. The AM pool comprises tissue‐resident AMs (TR‐AMs) and monocyte‐derived AMs (Mo‐AMs); yet how vaccination remodels the mixed AM pool for long‐term antimicrobial defense against multidrug‐resistant bacteria remains poorly understood. In this work, we applied intranasal inactivated whole‐cell (IWC) vaccination against Acinetobacter baumannii and systematically dissected AM population dynamics at cellular and molecular levels. Vaccination reshaped the AM pool by recruiting CD11B+CD13+ Mo‐AMs that gradually acquire a TR‐AMs‐like phenotype and training TR‐AMs via sustained transcriptional adaptations. Functionally, the IWC‐remodeled AM pool exhibited enhanced antigen presentation, TNF‐α secretion, and phagocytosis, accompanied by metabolic rewiring, thereby conferring durable protection against A. baumannii infection. ATAC‐seq revealed immune‐related chromatin remodeling and enrichment of transcription factors, including ETS, IRF, and bZIP family members. Similar AM pool remodeling was observed following immunization with IWC vaccines against Pseudomonas aeruginosa and Klebsiella pneumoniae, suggesting parallel innate responses across multiple common respiratory bacterial vaccines. Collectively, our study delineates the single‐cell transcriptional and epigenetic landscapes of vaccine‐remodeled AM subsets, providing mechanistic insights to guide the development of AM‐targeted prophylactic vaccines against multidrug‐resistant A. baumannii.

Gastric cancer remains a leading global health concern, with its etiology shaped by complex interactions between the host and its microbiome. The primary etiological role of Helicobacter pylori (H.pylori) has been well-established, but recent research has pointed to the significant contributions of non-H.pylori pathogens in the onset and progression of gastric cancer. These pathogens contribute to gastric tumorigenesis by directly compromising the gastric epithelial barrier and invading gastric epithelial cells, affecting long-range processes, disrupting microbial balance, and influencing the host's immune microenvironment. In the following, we comprehensively elucidated the potential mechanisms by which Streptococcus anginosus, Methylobacterium, Prevotella, Candida albicans, and Epstein-Barr virus actively participate in gastric tumorigenesis. Beyond this, ongoing investigations seek to identify additional microorganisms that may contribute to gastric cancer development, offering new insights into the multifactorial nature of the disease. Collectively, these findings highlight the critical involvement of diverse non-H.pylori microorganisms at various stages of gastric cancer progression, advancing our understanding of microbe-driven carcinogenesis.

BACKGROUND AND AIMS:We aimed to explore whether saxagliptin, a dipeptidyl peptidase-IV inhibitor, could ameliorate glucose fluctuations and maintain β-cell function in T1DM. METHODS AND RESULTS:A multicentre, open-label, randomised trial was performed, including 184 T1DM patients from six medical centres. These patients received insulin with or without saxagliptin at 5 mg per day for 24 weeks. The primary endpoint was the change from the baseline value of the MAGE, as measured by a CGMS after 24 weeks. The secondary endpoints included the change from baseline value of islet function during the 3-hour BMTT, HbA1c, and insulin dosage. The exploratory analysis was the influence of SNPs in the incretin-related genes on saxagliptin treatment outcomes. No differences were observed between the two groups in MAGE after treatment for 24 weeks. The change of C-peptidemax levels from baseline to 24 weeks in SAXA group (insulin plus saxagliptin) was higher than in CONT group (insulin only) [p = 0.040]. No difference were observed between the groups in HbA1c, insulin dosage after 24 weeks. In SAXA group, rs10305439, rs10305441 of GLP1R and rs6233 of PCSK1/3 were associated with HbA1c response (p = 0.026, 0.019, and 0.048 respectively); the G allele of rs2143734 of GLP1R were associated with lower change of fasting C-peptide from baseline (p = 0.029) CONCLUSIONS: The saxagliptin did not ameliorate glucose fluctuations; however, it appeared to maintain β-cell function to some extent, and SNPs in the incretin-related gene may indicate responsiveness to DPP-IV inhibitors in T1DM. CLINICALTRIALS:Gov number, NCT02307695.

BackgroundImmune checkpoint inhibitors (ICIs) have revolutionized cancer treatment. However, these therapies are associated with immune-related adverse events (irAEs), with thyroid dysfunction as one of the most common endocrine irAEs. The role of preexisting thyroid autoantibodies (Abs) in predicting endocrine irAEs and their impact on patient survival remains unclear.ObjectiveThis study explored the relationship between preexisting thyroid antibodies and the development of endocrine irAEs, as well as their association with overall survival (OS) in patients treated with ICIs.MethodsWe retrospectively reviewed patients with malignancy treated with one or more PD-(L)1 inhibitor between July 2018 and March 2022 at The First Affiliated Hospital of Nanjing Medical University, and 4,119 patients were initially included in this study. Ultimately, 585 patients whose anti-TPO and anti-Tg levels were measured before ICI treatment were analyzed. The patients were stratified based on their pre-existing thyroid antibody status. Multivariate Cox regression models were used to analyze the risk of thyroid irAEs and the differences in OS. Propensity score matching (PSM) and inverse probability of treatment weighting (IPTW) were employed to validate the robustness of the results.ResultsPatients with pre-existing thyroid Ab (N = 80, 13.7%) had a significantly higher incidence of thyroid irAEs compared to the antibody-negative group (N = 505) (HR = 4.13, 95% CI: 2.69-6.35, p < 0.001). PSM and IPTW analyses further confirmed that patients with pre-existing thyroid Ab had a markedly increased risk of grade≥2 irAEs (HR = 7.54, 95% CI: 2.74-20.75, p < 0.001). No significant difference in OS was observed between the two groups (log-rank p = 0.5). Notably, patients with pre-existing thyroid Abs and irAEs had better survival outcomes (log-rank p = 0.027). A landmark analysis at 12 months indicated that patients who developed thyroid irAEs between 0 and 12 months of age had relatively low mortality rates over this time period. However, from 12 months onwards, the survival curves of the two groups tended to overlap.ConclusionPreexisting thyroid antibodies provide a significant risk factor for thyroid irAEs in patients treated with ICIs. The development of thyroid irAEs has been associated with improved survival, thus suggesting a potential link between immune activation and clinical benefits.

BackgroundType 1 diabetes (T1D) is an autoimmune disease characterized by progressive β-cell destruction, yet current risk stratification tools, which rely mainly on genetic susceptibility and autoantibody profiles, remain insufficient for accurately predicting disease progression. We aimed to characterize macrophage-related inflammatory transcriptional activity in T1D and to develop peripheral blood–based biomarkers for diagnosis and risk stratification.MethodsWe integrated bulk RNA-seq, single-cell RNA-seq, and spatial transcriptomic data from human islets with public and in-house peripheral blood transcriptomic datasets. Macrophage heterogeneity and remodeling trajectories were analyzed in the islet microenvironment, and machine learning was used to derive tissue- and blood-based proinflammatory macrophage-related genes (PMRG). Diagnostic and prognostic models were then constructed and validated in peripheral blood cohorts, including a longitudinal islet autoimmunity (IA) cohort. SHAP analysis was applied to improve model interpretability. Independent PBMC RT-qPCR and mouse pancreatic immunofluorescence were performed to validate selected PMRG-related genes.ResultsT1D islets showed marked immune remodeling with myeloid enrichment and five distinct macrophage subtypes. Pseudotime analysis identified a pro-inflammatory macrophage trajectory and 265 PMRGs, from which a 9-gene islet-derived PMRG (iPMRG) was obtained. Spatial transcriptomics supported the association of iPMRG-high macrophage signals with disrupted β-cell regions, and CellChat analysis inferred altered inflammatory communication programs. In peripheral blood mononuclear cells (PBMCs), the iPMRG-based diagnostic classifier distinguished T1D from healthy controls with an optimism-corrected AUC of 0.736. For prognosis, a 15-gene prognostic PMRGs was used to construct a risk score that, when integrated with clinical variables, predicted progression from IA to clinical T1D with time-dependent AUCs of 0.825, 0.814, and 0.860 at 12, 36, and 60 months, respectively. SHAP analysis identified the PMRG risk score as the dominant predictor and highlighted six core driver genes (PID1, TFPI2, SERPINB2, SOX4, DUSP2, and MT1X). The computational findings were further supported by independent validation in PBMCs and mouse pancreatic tissues.ConclusionsOur study highlights the heterogeneous and dynamic nature of macrophage remodeling in the T1D islet microenvironment, which is translated into accessible peripheral blood signatures. The resulting diagnostic and prognostic models provide an interpretable framework for T1D risk stratification and may support future strategies for earlier detection and precision prevention.

Minoritized populations face many barriers to accessing evidence-based diabetes intervention. To evaluate the feasibility, acceptability, and potential efficacy of a social media-based intervention to improve glycemic control among Chinese Americans with type 2 diabetes. A pilot randomized controlled trial (RCT) with 3-month and 6-month follow-ups. Chinese Americans (n = 60, mean age 54.3 years old) with limited education (70.0 https://clinicaltrials.gov/ct2/show/NCT03557697

A proportion of patients with new-onset diabetes share similar symptoms with type 1 diabetes (T1D) patients but they are negative for islet antigen-specific autoantibodies. This study was to develop an islet antigen-specific CD8+ T-cell assay to provide autoimmune evidence regarding these “suspected” T1D patients. HLA-A*0201 individuals with autoAbs+ T1D, autoAbs− suspected T1D, and type 2 diabetes, along with HLA-A*0201 healthy controls were recruited. Using interferon-γ enzyme-linked immunospot assays, the percentages of participants in each group with various islet antigen-specific CD8+ T cells were determined. Sixteen out of the 28 islet antigen-specific epitopes tested were T1D specific, meaning that there was a significantly (P < 0.05) greater epitope positivity rate in the autoAbs+ T1D cohort than in the healthy controls. Using a cutoff value of two positive epitopes, the 16-epitope panel led to a sensitivity of 75.0

This study investigated the effect of Zygosaccharomyces rouxii (Z. rouxii) fermentation on the functional properties and flavor quality of Cynanchum auriculatum Royle ex Wight beverages (CAB). The findings revealed that Z. rouxii fermentation significantly altered the acid and sugar profiles, increased the ethanol content (P < 0.05), raised the total phenolic content by 11.50 %, and elevated the total flavonoid content by 60.30 %. The fermentation process also modified the polyphenol profiles, significantly affecting the levels of gallic acid, kaempferol, and gossypol (P < 0.05). Additionally, the antioxidant and alpha-glucosidase inhibitory activities were significantly enhanced (P < 0.05), particularly in ABTS (98.95 % inhibition) and DPPH (90.62 % inhibition) free radical scavenging abilities, as well as alpha-glucosidase inhibition (98.95 %). Analysis of volatile metabolites (VMs) identified 98 differential metabolites between fermented and non-fermented CAB, and 20 key flavor metabolites with a relative odor activity value (rOAV) of >= 1 were identified. Z. rouxii fermentation primarily changed the types and concentrations of alcohols, esters, and terpenoids in CAB, thereby enhancing its "floral, sweet, and woody" flavor profile. These findings provide valuable insights for biochemical research on fermented CAB.

BACKGROUND:Chinese Americans with type 2 diabetes (T2D) face many linguistic, cultural, and social determinants of health-related barriers to accessing evidence-based diabetes interventions. Our team developed the culturally and linguistically tailored Chinese American Research and Education (CARE) program to provide evidence-based diabetes education and support to this group and demonstrated the feasibility, acceptability, and potential efficacy of the intervention on improving hemoglobin A1c levels. However, it remains unclear whether the CARE program also improves diabetes self-efficacy and psychosocial outcomes in the same study sample. OBJECTIVE:This is a secondary analysis to examine the potential efficacy of the CARE program on secondary outcomes, including diabetes self-efficacy, self-care activities, beliefs in diabetes self-care activities, and diabetes distress among Chinese Americans with T2D. METHODS:A 2-arm, pilot randomized controlled trial was conducted to evaluate the CARE program between March 1, 2021, and April 21, 2023. The trial included 60 Chinese Americans aged 18 to 70 years who had a diagnosis of T2D and a baseline hemoglobin A1c level of 7% or higher. Participants were recruited from various health care settings in New York City, including community health centers, private primary care providers, and NYU Langone Health and its affiliates, and were randomly assigned to either the CARE intervention group (n=30) or a waitlist control group (n=30). The intervention consisted of 2 culturally and linguistically tailored educational videos per week for 12 weeks, covering diabetes self-care topics such as healthy eating, physical activity, and medication adherence. These videos were delivered via the WeChat app. In addition, community health workers provided support calls to assist them in setting goals, problem-solving, and addressing social determinants of health barriers every 2 weeks. Secondary outcomes included patient self-reported diabetes self-efficacy, self-care activities, beliefs in diabetes self-care activities, and diabetes distress. Outcomes were assessed at baseline, 3 months, and 6 months. RESULTS:Participants had a mean age of 54.3 (SD 11.5) years and 62% (37/60) were male, 78% (47/60) were married, 58% (35/60) were employed, 70% (42/60) had a high school education or lower, and 88% (53/60) reported limited English proficiency. Intervention participants demonstrated statistically significant improvements in self-efficacy at 3 months (estimated difference in change: 8.47; 95% CI 2.44-14.5; adjusted P=.02), diabetes distress at 6 months (estimated difference in change: -0.43; 95% CI -0.71 to -0.15; adjusted P=.009), and adherence to a healthy diet at both 3 months (estimated difference in change: 1.61; 95% CI 0.46-2.75; adjusted P=.02) and 6 months (estimated difference in change: 1.64; 95% CI 0.48-2.81; adjusted P=.02). CONCLUSIONS:The culturally and linguistically tailored intervention showed promise in improving self-efficacy and diabetes self-care activities among Chinese Americans with T2D, warranting validation through a large-scale randomized controlled trial. TRIAL REGISTRATION:ClinicalTrials.gov NCT03557697; https://clinicaltrials.gov/study/NCT03557697.

AIMS:To investigate the effects of different subtypes of lipohypertrophy (LH) on insulin total daily dose (TDD) requirements in patients with type 1 diabetes mellitus (T1DM), and to propose subtype-specific insulin dosage adjustment strategies after avoiding injection at sites of LH. METHODS:This prospective observational study enrolled hospitalised T1DM patients with a disease duration ≥1 year. Point-of-care ultrasound was performed immediately after their admission to determine the presence and the specific type of LH. An insulin pump was installed away from the LH sites of each patient. Continuous subcutaneous insulin was infused to control their blood glucose, and the insulin dose was titrated gradually until their blood glucose stabilised to the target. RESULTS:A total of 288 patients were included. According to ultrasound screening, 104 patients (36.11%) were LH free (LH-0) and 184 patients (63.89%) were found to have LH (LH+), of which 114 had nodular hyperechoic LH (LH-1), 62 had diffuse hyperechoic LH (LH-2) and 8 had hypoechoic LH (LH-3). Paired-sample t-test showed that all LH+ groups had a decrease in TDD and TDD/body weight after avoiding injections at the lesion sites (all p < 0.05). Compared with the LH-0 group, the TDD and TDD/body weight of the LH-2 and LH-3 groups were significantly reduced (all p < 0.05), whereas there was no significant change in the LH-1 group. Among the TDD reduced in the LH-2 and LH-3 groups, the bolus insulin dosage reduced accounted for the vast majority, with 88.05% (8.55/9.71 international unit [IU]/day) and 74.78% (18.12/24.23 IU/day), respectively. CONCLUSIONS:TDD reductions vary among patients with different subtypes of LH. Nodular hyperechoic LH may not require immediate dose adjustments. Patients with diffuse hyperechoic and hypoechoic LH necessitate bolus-focused dose reductions.

Tuberculosis, caused by Mycobacterium tuberculosis, is a severe and persistent global public health issue, particularly exacerbated by the emergence of multidrug-resistant and extensively drug-resistant strains. This study employed pan-genomic approaches to analyze different strains with various resistance profiles, examining the diversity of bacterial genetic evolution in relation to mutations in resistance-related genes. The findings indicate that resistance-related genes are mostly core genes (94%), with a preference for base mutations closely associated with nonsynonymous mutations at resistance sites. Interestingly, while the majority of drugs induce positive selection in target genes, the tlyA gene under the influence of amikacin (AMI) undergoes passive selection. Cluster analysis of target genes suggests consistency between SNP clusters and drug-resistant clusters, revealing a strong correlation between bacterial evolutionary branches and resistance profiles. Consequently, based on pan-genome evolutionary characteristics, we identified the drug-resistant mutation pattern (DRMP) that can serve as a molecular fingerprint and indicator for drug sensitivity, aiding in the assessment and guidance of drug selection for treating different strains and the formulation of individualized treatment plans. This research not only enhances our understanding of the mechanisms of drug resistance in M. tuberculosis but also offers new perspectives for the development of new drugs, which is crucial for global tuberculosis control.

BACKGROUND:We evaluated the effects of drug therapy on T lymphocyte subsets and their associations with recurrent chronic bronchitis (CB) attacks. METHODS:A total of 162 CB patients treated from April 2020 to April 2021 were selected. All patients underwent anti-infective, cough-relieving, and phlegm-eliminating treatment, as detailed in Clinical Pathway for Chronic Bronchitis. They were divided into a recurrent attack group (n=95) and a non-recurrent attack group (n=67). Changes in T lymphocyte subsets at different time points of treatment and their associations with the number of attacks were analyzed. Associated factors were analyzed in a multivariate logistic regression model, and their predictive value was validated using a nomogram prediction model and receiver operating characteristic (ROC) curves. RESULTS:The number of attacks at 1 year after treatment was positively correlated with CD8+ and negatively correlated with CD3+, CD4+, and CD4+/CD8+. A history of smoking, CD3+, CD4+, CD8+, and CD4+/CD8+ were independent risk factors for recurrent attacks. The nomogram prediction model showed that the total risk value corresponding to smoking history, low CD3+, CD4+, and CD4+/CD8+, and elevated CD8+ was 0.86, and the concordance index and area under the ROC curve of the model was 0.896 (95% CI: 0.782-0.997, P<0.05) and 0.816 (95% CI: 0.803-0.847, P<0.001), respectively. There was an association between T lymphocyte subsets and recurrent attacks before and after treatment of CB. CONCLUSIONS:Low CD3+, CD4+/CD8+ and CD4+, elevated CD8+, and smoking history were risk factors for recurrent attack.

This study described a case of plurihormonal tumor associated with papillary thyroid carcinoma (PTC) and summarized the treatment approaches for similar cases, while also exploring the underlying pathogenesis. The patient exhibited symptoms indicative of acromegaly, central hyperthyroidism, and hyperprolactinemia. A glucose loading test demonstrated persistently elevated growth hormone (GH) levels, while thyroid function tests revealed inappropriate thyroid stimulating hormone (TSH) secretion. Imaging of the pituitary gland revealed a 26 × 19 mm lesion compressing the optic chiasm. Thyroid ultrasound reveals bilateral Thyroid Imaging Reporting and Data System 4B nodules, with the largest on the right measuring 27 × 20 mm. Fine-needle aspiration cytology (FNAC) revealed the presence of PTC. Three weeks later, the patient underwent pituitary adenomectomy. Immunohistochemistry revealed a plurihormonal tumor positive for TSH, GH, luteinizing hormone (LH), prolactin (PRL), pituitary-specific transcription factor 1 (Pit1), and steroidogenic factor 1 (SF1). A total thyroidectomy followed 10 weeks post-adenomectomy. Immunohistochemical evaluation showed higher nuclear positivity for the high-mobility group AT-hook 1 (HMGA1) and the high-mobility group AT-hook 2 (HMGA2) proteins in neoplastic cells compared to normal tissues. In our search results, only three similar cases were identified, and we summarized the relevant literature search results which raise the possibility that the HMGA2-the Retinoblastoma Protein (pRB)/E2F Transcription Factor 1 (E2F1)-HMGA1 signaling pathway may represent a common pathogenic pathway for PTC and plurihormonal tumor. If a patient simultaneously suffers from PTC and plurihormonal tumor, the determination of the surgical sequence is crucial. In cases of postoperative recurrence, where patients are reluctant to undergo additional surgeries, targeting HMGA is likely to offer a promising approach to prevent the progression of both the pituitary tumors and PTC.

ObjectiveTo analyze the clinical characteristics of immune checkpoint inhibitor (ICI)-related hypophysitis. MethodsA retrospective analysis was conducted on patients diagnosed with ICI-related hypophysitis and treated at the Department of Endocrinology, The First Affiliated Hospital with Nanjing Medical University, between January 2020 and March 2025. Clinical manifestations and prognosis of patients were analyzed. ResultsEleven patients with ICI-related hypophysitis were included. The average age was (62.27±7.63) years, and 9 patients (81.82%) were male. The median time to onset was 9.1 months, and the median number of treatment cycles received was 5. The primary initial symptoms were fatigue and anorexia. Hyponatremia was present in 3 patients (27.27%). Evaluation of anterior pituitary function revealed adrenocorticotropic hormone deficiency as the most common manifestation (90.91%, 10/11), followed by hyperprolactinemia (81.82%, 9/11). Posterior pituitary function remained normal in all patients. Pituitary magnetic resonance imaging showed no abnormality in 4 patients (44.44%). Thyroid dysfunction was observed in 6 patients (54.55%), one of whom (9.09%) also exhibited pancreatic endocrine dysfunction. The average follow-up duration was 36.5 months. Eight patients (72.73%) were alive at the last follow-up. None of the patients recovered their pituitary hormone function. ConclusionsEndocrine adverse events induced by ICIs can involve multiple glandular systems. Clinicians should be highly vigilant for the possibility of ICI-induced hypophysitis in patients receiving ICIs who present with symptoms such as fatigue, anorexia, and hyponatremia.

Objective The aim of this retrospective study was to investigate the relevance of influenza A virus (IAV) in acutely exacerbating airway inflammatory response and disrupting immune function in elderly COPD patients. Methods The group conducted a pre-test: using multiplex combined real-time PCR detection kits Multiple real⁃time PCR was used to detect twenty-four pathogens, 385 patients clinically diagnosed with COPD were tested for viral nucleic acid in throat swabs. At the same time, peripheral blood leukapheresis was collected from both groups of patients, and their IL-6, IL-8, IL-1β, and TNF-α levels were detected, along with the levels of T-cell differentiation markers CD4 and CD8, to assess the influence of influenza virus on the immune function of elderly COPD patients and its relevance to the acute exacerbation of airway inflammatory response in elderly COPD patients. Results Results showed that the expression of inflammatory cytokines IL-6, IL-8, IL-1β and TNF-α was significantly higher in the viral group compared with the non-infected group (P < 0.05, P < 0.01). The levels of T cell differentiation type markers CD4 and CD8 were significantly lower in the infected group compared with the uninfected group. Conclusion Influenza virus further exacerbated airway inflammatory response and decreased the immune function of T cells by activating intrinsic immune molecules such as IL-6, IL-8, IL-1β and TNF-α.

ABSTRACT Drug efflux systems have recently been recognized as a significant mechanism responsible for multidrug resistance in bacteria. In this study, we described the identification and characterization of a new chromosomally encoded efflux pump (SA00565) in Staphylococcus aureus . SA00565, which belongs to the drug/metabolite transporter (DMT) superfamily, was predicted to be a 10-transmembrane segment transporter. To evaluate the role of sa00565 in resistance, we generated sa00565 gene deletion mutant (Δ sa00565 ) and assessed its susceptibility to 35 different antibiotic treatments. Our results demonstrated that the Δ sa00565 mutant exhibited reduced resistance to tetracycline and doxycycline, with 64-fold and 12-fold decreased MICs, respectively. The mechanism of SA00565-mediated tetracycline resistance was demonstrated that SA00565 possesses the capability to efficiently extrud intracellular tetracycline into the environment. The efflux activity of SA00565 was further validated using EtBr accumulation and efflux assays. In summary, our study uncovered a previously unknown function of a DMT family transporter, which serves as a tetracycline efflux pump, thereby contributing to tetracycline resistance in S. aureus . IMPORTANCE In this study, we addressed the significance of drug efflux systems in multidrug resistance of Staphylococcus aureus , focusing on the unexplored efflux pump SA00565 in the drug/metabolite transporter (DMT) superfamily. Through phylogenetic analysis, gene knockout, and overexpression experiments, we identified the role of SA00565 in antibiotic resistance. The Δ sa00565 mutant showed increased susceptibility to tetracycline and doxycycline in disk diffusion assays, with significantly lower MICs compared to the WT. Remarkably, intracellular tetracycline concentration in the mutant was two- to threefold higher, indicating SA00565 actively eliminates intracellular tetracycline. Our findings emphasize the pivotal contribution of SA00565 to tetracycline antibiotic resistance in S. aureus , shedding light on its functional attributes within the DMT superfamily and providing valuable insights for combating multidrug resistance.

Abstract Background A subset of Graves’ disease (GD) patients develops refractory hyperthyroidism, posing challenges in treatment decisions. The predictive value of baseline characteristics and early therapy indicators in identifying high risk individuals is an area worth exploration. Methods A prospective cohort study (2018–2022) involved 597 newly diagnosed adult GD patients undergoing methimazole (MMI) treatment. Baseline characteristics and 3-month therapy parameters were utilized to develop predictive models for refractory GD, considering antithyroid drug (ATD) dosage regimens. Results Among 346 patients analyzed, 49.7% developed ATD-refractory GD, marked by recurrence and sustained Thyrotropin Receptor Antibody (TRAb) positivity. Key baseline factors, including younger age, Graves’ ophthalmopathy (GO), larger goiter size, and higher initial free triiodothyronine (fT3), free thyroxine (fT4), and TRAb levels, were all significantly associated with an increased risk of refractory GD, forming the baseline predictive model (Model A). Subsequent analysis based on MMI cumulative dosage at 3 months resulted in two subgroups: a high cumulative dosage group (average ≥ 20 mg/day) and a medium–low cumulative dosage group (average < 20 mg/day). Absolute values, percentage changes, and cumulative values of thyroid function and autoantibodies at 3 months were analyzed. Two combined predictive models, Model B (high cumulative dosage) and Model C (medium–low cumulative dosage), were developed based on stepwise regression and multivariate analysis, incorporating additional 3-month parameters beyond the baseline. In both groups, these combined models outperformed the baseline model in terms of discriminative ability (measured by AUC), concordance with actual outcomes (66.2% comprehensive improvement), and risk classification accuracy (especially for Class I and II patients with baseline predictive risk < 71%). The reliability of the above models was confirmed through additional analysis using random forests. This study also explored ATD dosage regimens, revealing differences in refractory outcomes between predicted risk groups. However, adjusting MMI dosage after early risk assessment did not conclusively improve the prognosis of refractory GD. Conclusion Integrating baseline and early therapy characteristics enhances the predictive capability for refractory GD outcomes. The study provides valuable insights into refining risk assessment and guiding personalized treatment decisions for GD patients.