Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.

Objective:Primary pulmonary mucoepidermoid carcinoma(PMEC)is a rare malignant lung tumor that accounts for approxim-ately 0.1%-0.2%of all primary pulmonary neoplasms.Due to the non-specific clinical symptoms and epidemiological features,PMEC poses diagnostic challenges.Methods:Tissue blocks from 23 archived PMECs were collected from The First Affiliated Hospital of Soochow Uni-versity(November 2012 to December 2023).To establish definitive diagnoses,comprehensive histopathological evaluation,including histo-morphological analysis,immunohistochemistry(IHC),fluorescence in situ hybridization(FISH),and periodic acid-Schiff(PAS)staining were performed.Results:The tumors consisted of varying proportions of mucin-secreting cells(mucous cells),intermediate cells,and epidermoid cells.Immunophenotypically,CK7 was predominantly expressed in the mucous cells,whereas CK5/6,p40,and p63 were expressed in the epidermoid and intermediate cells.The Ki-67 proliferation index ranged from 5%to 60%.All tumors were negative for TTF-1 and Napsin A.Five of the tumors were positive for PD-L1(clone 22C3),with a tumor percentage score of 3%-20%.All 11 tumors tested for ALK(clone D5F3)were negative.IHC for c-Met was performed on two tumors and both were weakly positive(+).Mastermind-like transcriptional co-activator 2(MAML2)gene rearrangement was detected in 34.8%(8/23)of the tumors.Mucous cells were PAS positive.Kaplan-Meier surviv-al analysis revealed a significantly poorer prognosis for patients with lymph node metastasis,distant metastasis,advanced TNM stage(Ⅲ+Ⅳ),poor differentiation,or MAML2 gene rearrangement negativity.Univariate analysis identified poor histological differentiation,lymph node metastasis,distant metastasis,and advanced TNM stage as the major prognostic risk factors.Multivariate analysis confirmed poor differentiation and distant metastasis as independent risk factors for adverse outcomes.Conclusions:PMEC is an aggressive tumor with low incidence and non-specific clinical manifestations,leading to frequent misdiagnosis.Clinicians should maintain a high index of suspicion and ensure a thorough differential diagnosis.

Objective To observe the clinical and transthoracic echocardiographic manifestations of mitral annular disjunction(MAD).Methods Transthoracic echocardiographic data of 990 individuals were retrospectively analyzed.Based on the distance between the attachment point of posterior mitral valve and posterior wall of left ventricle measured on echocardiography,whether there was MAD was judged,and the subjects were divided into MAD group and non-MAD(NMAD)group,and clinical data and echocardiographic parameters were compared between groups.Taken parameters being significantly different between groups as independent variables and the presence or absence of MAD as dependent variable,multivariate binary logistic regression analysis was used to explore correlations of patients' clinical characteristics and transthoracic echocardiographic parameters of MAD.Results MAD was detected in 186 cases(186/990,18.79％).Patients' age,left atrial diameter(LAD),left ventricle diameter(LVD),late diastolic velocity of mitral valve(A)and early diastolic velocity of mitral valve orifices(E)/A in MAD group were all lower than those in NMAD group,while the proportion of moderate and above mitral regurgitation in MAD group was higher than that in NMAD group(all P＜0.05).Patients' age,A and E/A measured with transthoracic echocardiography were all correlated with MAD(all P＜0.05).Conclusion Clinical and transthoracic echocardiographic manifestations of MAD had certain characteristics.

Objective:To retrospectively analyze the efficacy and safety of different radiotherapy doses in treating esophageal squamous carcinoma(ESCC)patients aged ≥80 years.Methods:This retrospective study collected clinical data from ESCC patients aged ≥80 years who underwent radiotherapy at the Fourth Hospital of Hebei Medical University from January 2016 to September 2021.Observation variables included overall survival(OS), progression free survival(PFS), complete response(CR), partial response(PR), stable disease(SD), progressive disease(PD), and adverse reactions.Survival rates were estimated using the Kaplan-Meier method and compared via log-rank tests.Cox regression models were employed for multivariate analysis.Results:A total of 165 patients who met the enrollment criteria were included in this study, including 88(88/165, 53.3%)males and 77(77/165, 46.7%)females, age 80 to 100 years(median age 83 years), 66 cases(66/165, 40.0%)in the radiotherapy dose ≥60 Gy group and 99 cases(99/165, 60.0%)in the <60 Gy group.Of the 165 patients, the effective rate was 71.5%(118/165).The median overall survival(OS)of the whole group was 19.0 months, The median Progression Free Survival(PFS)for the whole group was 13 months.The results showed that radiotherapy dose, lesion length, Nutritional Risk Index(NRI), eating condition, recurrence/progression and chemotherapy were factors influencing OS.The survival of patients in the ≥60 Gy group, ≤5 cm group, well-nourished group(NRI ≥45)group, soft diet and general diet group and combined chemotherapy group is better.Cox multivariate analysis revealed that radiotherapy, dose eating condition and the lesion length were independent prognostic factors for OS.The OS rate of the radiation therapy group with a dose of ≥ 60 Gy was better than that of the<60 Gy group( P=0.001), the OS of the general or soft diet group was better than that of the semi liquid or liquid diet group( P=0.008), and the OS of the lesion length ≤ 5 cm group was better than that of the>5 cm group( P=0.020).The incidence rates of radiation-induced esophagitis, myelosuppression, radiation pneumonia, and gastrointestinal reactions in the entire group were 60.0%(99/165), 12.1%(20/165), 22.4%(37/165), and 14.5%(24/165), respectively.51.5%(85/165)of the group experienced local recurrence, 10.3%(17/165)had distant organ metastasis, and 9.1%(15/165)had non regional lymph node metastasis.As of the follow-up date, there were a total of 99 deaths in the entire group. Conclusions:For patients aged 80 years or older with esophageal cancer, higher radiation doses, better feeding and nutritional status have more beneficial for prolonged survival.Local recurrence remains the main reason for treatment failure in elderly patients with esophageal cancer.

Objective To screen the influencing factors associated with adverse clinical out-comes in hospitalized elderly patients with Alzheimer's disease(AD)using LASSO-Logistic regression analysis and to construct a nomogram prediction model.Methods A retrospective selection of 214 hospitalized elderly patients with AD who visited the Department of Geriatric Medicine in the hospital from February 2021 to March 2023 was conducted,and clinical data of all patients were collected.Patients were divided into adverse events group(n=53)and non-adverse events group(n=161)based on the occurrence of adverse clinical outcomes.After variable screening using LASSO regres-sion,multivariate Logistic regression analysis was performed to identify independent factors influen-cing adverse clinical outcomes in hospitalized elderly patients with AD.A Nomogram model for pre-dicting adverse clinical outcomes in these patients was established based on the results of multivariate analysis.The predictive performance,calibration,and clinical utility of the Nomogram model were evaluated using the concordance index,calibration curve,and decision curve analysis(DCA).The diagnostic performance of the Nomogram model for adverse clinical outcomes in hospitalized elderly patients with AD was assessed using the receiver operating characteristic(ROC)curve and the area under the curve(AUC).Results LASSO-Logistic regression analysis revealed that the Mini-Men-tal State Examination(MMSE)score was an independent protective factor against adverse clinical outcomes in hospitalized elderly patients with AD(P＜0.05),while the Charlson Comorbidity In-dex(CCI score),creatinine,urea,and fasting blood glucose(FBG)levels were all independent risk factors for adverse clinical outcomes in these patients(P＜0.05).The Nomogram model con-structed based on the influencing factors screened by LASSO-Logistic regression analysis showed a concordance index of 0.994(95％CI,0.958 to 1.000)for predicting adverse clinical outcomes in hospitalized elderly patients with AD.The Hosmer-Lemeshow test results indicated x2=1.909,P=0.984,suggesting good model fit.The DCA result demonstrated that the model had favorable threshold probabilities and net clinical benefits.Conclusion The Nomogram model for predicting clinical outcomes in elderly inpatients with AD constructed based on LASSO-Logistic regression anal-ysis exhibits high predictive value,and can be used to forecast the occurrence of adverse clinical outcomes in these patients.

Objective To establish models for real-time dynamic monitoring of intracellular cytoplasmic adenosine triphosphate(ATP)and mitochondrial ATP levels in cells in order to study the changes in metabolic processes.Methods The lentiviral plasmids of the cytoplasmic chemogenetic green fluorescent protein(GFP)ATP probe(Chemo-G)and those of the mitochondrial-localized chemogenetic GFP ATP probe(mito-Chemo-G)were constructed before being transfected into HEK293T together with the helper plasmids,respectively.Chemo-G and mito-Chemo-G lentiviruses were obtained.HeLa cells were infected with the lentivirus.Puromycin resistance selection and flow cytometry cell sorting were employed to identify and isolate the infected HeLa cells.The growth and GFP expressions of HeLa cells were observed.A live-cell imaging system was used for continuous imaging of the cells,with stimuli added at specific time points to alter intracellular ATP levels to observe changes in the fluorescence intensity of the ATP probe.Results Lentiviral plasmids containing Chemo-G and mito-Chemo-G sequences were constructed.Two cell lines which could stably express Chemo-G and mito-Chemo-G were established that exhibited strong growth and accurate intracellular fluorescence localization.Live-cell imaging revealed that after the addition of 2-deoxy-D-glucose(2-DG)into HeLa-Chemo-G,the fluorescence resonance energy transfer(FRET)/GFP ratio showed a decrease that was partially reversed by the addition of glucose.The FRET/GFP ratio increased after histamine stimulation,but rapidly decreased after the addition of oligomycin.Conclusion The Chemo-G and mito-Chemo-G lentiviral vectors and stably transfected cell lines HeLa-Chemo-G and HeLa-mito-Chemo-G are constructed,which provides reliable experimental models for studying cellular metabolism and changes in intracellular ATP levels.

Objective To establish a stable in vitro model of CD8+T cell exhaustion.Methods CD8+T cells were isolated and purified from the spleens of ovalbumin-specific CD8+T cell receptor(OT-I)transgenic mice and subjected to chronic antigen stimulation to induce exhaustion in vitro.Flow cytometry was employed to evaluate the expressions of exhaustion markers,secretion of effector cytokines,and transcription factor profiles in CD8+T cells.Exhausted and effector(non-exhausted)CD8+T cells were co-cultured with tumor cells before tumor cell viability was measured to assess the cytotoxic potential of CD8+T cells.Additionally,N-acetyl-L-cysteine(N-AC)was used as a positive control during exhaustion induction to validate the model.Results Chronic stimulation resulted in a significant upregulation of inhibitory receptors,including programmed cell death protein 1(PD-1),T cell immunoglobulin and mucin domain-containing protein 3(TIM-3),T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motifdomain(TIGIT),and lymphocyte activation gene 3(LAG-3).Concurrently,the secretion of key effector cytokines such as tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)was markedly reduced.Exhausted CD8+T cells exhibited diminished cytotoxicity against tumor cells compared to effector CD8+T cells.Notably,treatment with N-AC effectively restored the function of exhausted CD8+T cells and enhanced their anti-tumor activity.Conclusion This study has established an effective in vitro model for CD8+T cell exhaustion.The use of N-AC demonstrates its potential to restore functionality in exhausted CD8+T cells,underscoring the reliability and utility of this model for investigating the anti-tumor potential of exhausted T cells.

Pancreatic acinar cell carcinoma (PACC) is a rare exocrine tumor of the pancreas with distinct clinical and pathological features. In recent years, advancements in molecular biology techniques have led to a deeper understanding of the molecular mechanisms underlying PACC. Progress in imaging, endoscopic, and molecular diagnostic technologies has improved the early detection rate of PACC. The primary treatment modalities for PACC include surgical resection, chemotherapy, targeted therapy and immunotherapy; however, the therapeutic efficacy still requires further improvement. This article reviews the current research status of PACC, covering its epidemiology, pathological characteristics, molecular alterations, diagnostic methods, and treatment strategies, and discusses the controversies and future directions in PACC research.

Sustained inflammatory responses are closely related to various severe diseases,and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment.Natural products have garnered considerable concern for the treatment of inflammation.Huanglian-Wumei decoction(HLWMD)is a classic prescription used for treating inflammatory diseases,but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated.Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory,we successfully obtained berberine(BBR)-chlorogenic acid(CGA)supramolecular(BCS),which is an herbal pair from HLWMD.Using a series of characterization methods,we confirmed the self-assembly mechanism of BCS.BBR and CGA were self-assembled and stacked into amphiphilic spherical supra-molecules in a 2:1 molar ratio,driven by electrostatic interactions,hydrophobic interactions,and π-πstacking;the hydrophilic fragments of CGA were outside,and the hydrophobic fragments of BBR were inside.This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules.Compared with free molecules,BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide(LPS)-induced pyroptosis.Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB(NF-κB)p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.

Objective To estimate the biological dose in a patient who developed radiation dermatitis after a local X-ray exposure incident. Methods Peripheral blood samples, which were used to performed lymphocyte chromosome aberration analysis, were collected from the patient at 54 and 102 days after the last exposure. Biological dose in the patient was estimated using four published X-ray dose-effect calibration curves for chromosomal aberrations. The absorbed dose in the patient was reconstructed using Dolphin′s model and time correction factors. Results The abnormal rates of chromosome aberration at 54 and 102 days after exposure were 1.00% and 0.40%, respectively. Based on the four calibration curves, the estimated local exposure dose at 54 day ranged from 3.59 to 10.51 Gy, and the time-corrected whole-body equivalent dose ranged from 0.27 to 0.87 Gy. The local dose estimated at 102 days ranged from 2.24 to 6.64 Gy, with a time-corrected whole-body equivalent dose of 0.12 to 0.60 Gy, which differed from the day-54 estimates. The biological doses estimated by both methods were lower than the physical dose (29.43 Gy). Conclusion The estimation of local biological dose of patient various in four dose-effect curves selected in this study. Delayed blood sampling will lead to underestimate biological dose. Early blood collection after radiation incidents is critical to ensure accuracy and reliability. Moreover, biological dose reconstruction methods for complex exposure scenarios require further research to improve the accracy of emergency response in radiation accidents.