Objective·To investigate the impact of KH-type splicing regulatory protein(KHSRP)on the proliferation of prostate cancer cells and the regulation of downstream gene expression,and explore the potential role and mechanism of KHSRP in the transition of prostate cancer from androgen-dependent to androgen-independent.Methods·Recombinant lentivirus was used to infect androgen-dependent prostate cancer LNCaP cells and androgen-independent prostate cancer DU 145 cells to establish stable cell lines with functional deficiency/acquisition of KHSRP,and the functional differences of KHSRP between the two cell types were compared.Western blotting was used to detect the expression levels of KHSRP,androgen receptor(AR),and ankyrin 3(ANK3)in the stable cell lines.Cell proliferation assay,colony formation assay,and mouse xenograft assay were performed to assess the impact of KHSRP on the proliferation ability of LNCaP cells.RNA sequencing(RNA-seq)was performed to identify downstream genes affected by KHSRP,and the mRNA expression levels of these genes were measured by quantitative real-time PCR(RT-qPCR).The expression of ANK3 and KHSRP in prostate tissue samples and the difference of ANK3 expression in different prostate cancer cell lines were analyzed by combining Cancer Cell Line Encyclopedia(CCLE),The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)databases.Results·GEO data analysis showed that KHSRP was upregulated in prostate cancer tissues compared to benign prostate tissues,suggesting its association with prostate tumorigenesis.Cell proliferation assay,colony formation assay,and mouse xenograft assay demonstrated a negative correlation between KHSRP expression and the proliferation ability of LNCaP cells,indicating that KHSRP can inhibit the proliferation of prostate cancer cells,with a stronger effect on LNCaP cells than on DU 145 cells.Western blotting and RT-qPCR analysis of the stable LNCaP cell lines showed that KHSRP overexpression led to a decrease in AR protein level without affecting its mRNA level,suggesting that KHSRP can indirectly regulate AR protein level.RNA-seq and RT-qPCR results showed KHSRP was positively correlated with the expression of ANK3,a regulatory factor affecting AR protein stability,and subsequent Western blotting confirmed an increase in ANK3 protein expression after KHSRP overexpression.TCGA data analysis further supported the correlation between KHSRP and ANK3 mRNA expression.Interestingly,according to CCLE and GEO data,the expression of ANK3 was closely related to prostate cancer malignancy.Conclusion·KHSRP may indirectly regulate AR protein stability through ANK3,thereby influencing the proliferation of androgen-dependent prostate cancer cells and mediating the altered responsiveness to androgen in prostate cancer cells.

Objective·To analyze the expression of mitochondrial related gene SFXN3(sideroflexin 3)in head and neck squamous cell carcinoma(HNSCC)and its effect on cell proliferation.Methods·Mitochondrial related genes and TCGA-HNSCC dataset were obtained from public databases to identify the target gene SFXN3 by using bioinformatic methods.The expression of SFXN3 in HNSCC patient samples was analysed by using the UALCAN database,and survival analyses of patients with different SFXN3 expression levels were performed according to TCGA-HNSCC cohort and GEO cohorts(GSE65858,GSE41613 and GSE27020).By using TCGA-HNSCC cohort and GEO cohorts(GSE40020,GSE210287),the differences in SFXN3 expression levels between therapeutic responders and non-responders were compared.Quantitative real-time PCR(qRT-PCR)was used to verify the expression of SFXN3 in the collected HNSCC tumor and para-tumor tissues,and the expression level of SFXN3 in human normal oral epithelial cells and HNSCC tumor cell lines was detected.With the construction of stable-SFXN3-knockdown HNSCC cell lines,the effect of SFXN3 on the proliferation ability of HNSCC cells was observed by using the Incucyte live-cell imaging analysis system and plate colony formation assay.Transcriptome sequencing was performed on the total RNA of stably-SFXN3-knockdown cells and control cells,and pathway enrichment analysis was performed on the genes with differentially down-regulated expression in knockdown group compared with control group.Results·SFXN3 was highly expressed in tumor tissues of HNSCC patients(P=0.000),and the prognosis of patients in the SFXN3-high-expression group was poor(all P＜0.05).The expression level of SFXN3 in the non-responders was higher than that in the responders(P=0.008),indicating an unfavorable prognosis.High expression of SFXN3 was verified in the collected HNSCC tumor tissues and HNSCC cell lines(all P＜0.05).After the knockdown of SFXN3 expression,the proliferation ability of HNSCC cells decreased,and the number of plate clones decreased(all P＜0.05).RNA sequencing showed that the differentially expressed down-regulated genes in HNSCC cells in the SFXN3-knockdown group were enriched in DNA replication,cell cycle,mitochondrial translation,mitochondrial RNA metabolic process and mitochondrial gene expression pathways.Conclusion·SFXN3 is highly expressed in HNSCC and it has negative correlation with the prognosis of patients.The proliferation ability and plate colony formation ability are inhibited in SFXN3-knockdown HNSCC cells,and these may work by affecting mitochondria function.

Objective·To establish a patient-derived tumor xenograft(PDX)model using tumor cells sourced from malignant pleural effusion(MPE)in patients with lung cancer,and to conduct experimental validation.Methods·Gene expression data were downloaded from the Gene Expression Omnibus(GEO),including single-cell RNA sequencing data for lung cancer with MPE(GSE131907)and for solid lung cancer(GSE203360).Data were clustered,and differential gene ontology functional enrichment analysis was performed to ascertain the feasibility of modeling by using MPE.MPE samples from patients with lung cancer were collected and processed through centrifugation and red blood cell lysis to enrich cells.The enriched cells were then implanted subcutaneously into non-obese diabetic/severe combined immunodeficient(NOD/SCID)mice.Tumor growth was monitored,and when tumors reached 1 000 mm3,they were passaged and preserved.Histopathological examination was conducted on stable passaged tumors,the cell morphology was observed via hematoxylin-eosin(H-E)staining and the expression of lung cancer biomarkers was detected by using immunohistochemistry(IHC).Results·Single-cell data analysis revealed stronger proliferative functions of tumor cells in MPE,suggesting that PDX modeling using MPE tumor cells may yield better tumor formation.A total of 35 samples of MPE from lung cancer patients were collected,and 13 PDX models were successfully constructed,with a success rate of 37.14％.Histopathological examination showed significant cellular atypia by H-E staining,and IHC result showed positive expression of lung cancer biomarkers such as cytokeratin 7(CK7),thyroid transcription factor-1(TTF1),and Napsin A.Conclusion·By enriching tumor cells from MPE of lung cancer patients,a more convenient,efficient,and dynamically modelable PDX model is successfully constructed.This model retains the malignant characteristics and protein expression features of tumor cells from lung cancer patients,providing an important experimental model tool for basic research and clinical drug guidance for lung cancer patients with MPE.

Objective·To investigate the evolution of sarcoma antigen 1(SAGE1),a member of the cancer-testis antigen(CTA)family,in three representative primate species—Macaca mulatta(Rhesus),Callithrix jacchus(Marmoset),and Microcebus murinus(Mouse Lemur),as well as the conservation of its interaction with INTS3,a subunit of the integrator complex.Methods·The interaction between INTS3 and SAGE1 in these primates and the interaction between INTS3 and human SAGE1 mutants were first validated in HEK293T cells by using co-immunoprecipitation(Co-IP).Truncated proteins were then tagged with His-SUMO and GST,respectively,and expressed in Escherichia coli,followed by a series of purification steps to obtain the target proteins.Surface plasmon resonance(SPR)was used to measure the binding affinity of the target proteins,and size exclusion chromatography with multi-angle light scattering(SEC-MALS)was used to determine the stoichiometry of the complex.Additionally,molecular docking was employed to predict the binding model of the truncated SAGE1 and INTS3.Mutations were performed on human SAGE1 key interface residues to analyze their binding to INTS3 by Co-IP.Results·The interaction between the full-length human-derived INTS3 and the full-length SAGE1 from the three primate species was confirmed by Co-IP.Truncated proteins were purified through multiple steps of tandom chromatography.The dissociation constants of the three pairs of truncated INTS3-SAGE1 were measured via SPR,and the SEC-MALS results demonstrated that the binding ratio of INTS3-SAGE1 was 2∶1.Furthermore,molecular docking predicted a structural model of the truncated INTS3-SAGE1 complex and the binding interface was extensively constituted with hydrophobic contacts assisted by some hydrophilic interactions.The interaction between the mutant SAGE1 and INTS3 full-length protein was significantly weakened.Conclusion·There is a conserved interaction between INTS3 and SAGE1 across Rhesus,Marmoset,and Mouse lemur.

Objective·To investigate the pathological and physiological changes underlying noise-induced cochlear nucleus damage and the regulating function of astrocytes on the damage,using a combination of morphological analysis,and molecular biology techniques.Methods·Forty-eight male C57BL/6J mice were randomly divided into two groups and exposed to 110 dB SPL(sound pressure level)broadband noise for 2 hours.Auditory brainstem response(ABR)tests were performed on the mice on days 1,7,14,30,and 90 after the noise exposure.Immunofluorescence staining of cochlear nuclear tissue was conducted to observe cochlear nuclear neurons and auditory synapses,as well as astrocyte activation levels.In addition,the damage to the cochlear nuclear neurons and synapses caused by noise was verified through Western blotting.Results·A significant decrease in cochlear nuclear Bushy cells after noise exposure was observed.The Western blotting results showed that there was severe loss of nerve fibers in cochlear nuclear neurons,indicating that noise caused significant damage to cochlear nucleus neurons.Moreover,a significant loss of auditory synapses labeled with vesicular glutamate transporter 1(Vglutl)was observed,which was the severest on day 14 after noise exposure and slowly recovered on day 90.Interestingly,astrocytes in the cochlear nucleus displayed obvious clustering and activation after noise exposure.By staining with glial fibrillary acidic protein(GFAP),most astrocytes were distributed around the cochlear nucleus,granule cell area,and auditory nerve root before noise exposure,and they had a small size.However,on day 14 after noise exposure,a large number of activated astrocytes aggregated in the ventral cochlear nucleus,and they all showed a pattern of growth around the synapses.Conclusion·Noise exposure leads to significant damage in the cochlear nucleus,and it is possible that astrocytes are involved in its damage and repair processes.These findings will provide a crucial foundation for further understanding the mechanisms of sound signal analysis,integration,and neural plasticity in the cochlear nucleus.

Objective·To investigate the current situation and distribution characteristics of chronic comorbidities,and to provide reference for further improving the self-management of comorbidities and implementing the whole course and all-round management of comorbidity.Methods·Two thousand and forty-five inpatients in the Department of Geriatrics,Renji Hospital,Shanghai Jiao Tong University School of Medicine were enrolled in this study from December 2020 to February 2023.The general vital signs,routine laboratory examination and disease status were collected.The epidemiological distribution characteristics of chronic diseases and comorbidities were analyzed.Results·The incidence of chronic diseases in the surveyed population was 99.6％,and the incidence of comorbidities was 94.2％.The top 5 chronic diseases were hypertension(43.68％),diabetes mellitus(24.81％),malignant tumor(21.48％),hyperlipidemia(18.38％)and coronary heart disease(11.99％).The detection rates of hypertension,diabetes mellitus,coronary heart disease,chronic obstructive pulmonary disease,stoke and chronic kidney disease in males were significantly higher than those in females(P＜0.05).The proportion of patients with 5 chronic diseases was the highest(11.99％),followed by 7 chronic diseases(10.26％)and 6 chronic diseases(10.04％).Among the patients of different ages,the comorbidity rate was the highest in the patients aged 50-59 years(27.78％).In different age groups,patients aged 50 to 59 with 2 chronic diseases had the highest incidence of comorbidity,which was as high as 40.82％.Although the overall proportion of comorbidities among male patients(95.37％)was higher than that among females(93.77％),there was no statistically significant difference(P=0.125).However,the proportions of male patients with 2 and 5 chronic diseases were 70.41％and 60.63％,respectively,which were significantly higher than those of female patients(29.59％and 39.37％).The correlations between coronary heart disease and diabetes mellitus,hypertension and coronary heart disease,hypertension and diabetes mellitus were higher(r=0.24,r=0.27,r=0.35,all P＜0.05).Conclusion·The prevalence of chronic diseases and comorbidities is high in the middle-aged and elderly population,and the number of comorbidities increases significantly with the increase of age.

Objective·To explore the value of miR-499 expression levels in the serum of patients with acute coronary syndrome(ACS)in predicting the severity of coronary artery disease.Methods·Seventy-four patients with ACS admitted to Xinhua Hospital,Shanghai Jiao Tong University School of Medicine were included and divided into a mild group of 30 cases,a moderate group of 21 cases,and a severe group of 23 cases according to the Gensini score.Real-time quantitative polymerase chain reaction(qPCR)technology was used to detect the expression level of serum miR-499,while troponin(cTnI),creatine kinase isoenzyme(CK-MB),myoglobin(Mb),brain natriuretic peptide(BNP),left ventricular ejection fraction(LVEF),and other laboratory indicators were measured.Spearman correlation analysis was used to analyze the correlation between the expression level of miR-499 and Gensini score,as well as the number of vessel branches in coronary artery disease.The evaluation value of miR-499 in assessing the severity of coronary artery disease was analyzed by using receiver operating characteristic(ROC)curve.Results·The expression level of miR-499 increased with the severity of coronary artery disease(P=0.000).The expression level of miR-499 was positively correlated with Gensini score and cTnI(r=0.331,r=0.143,P＜0.05),respectively,but not with LVEF(P＞0.05),while Gensini score was not correlated with cTnI and LVEF(P＞0.05).ROC analysis showed that the AUC of miR-499 for assessing the severity of coronary artery disease was 0.802(95％CI 0.698-0.906),with sensitivity of 65.2％and specificity of 80.4％.The expression level of miR-499 increased with the increase of the number of coronary artery lesions,and was positively correlated with the number of coronary artery lesions(r=0.368,P＜0.05).Conclusion·The expression level of miR-499 in the serum of ACS patients is correlated with the severity and number of coronary artery lesions,which has a certain evaluation value in evaluating the severity of coronary artery lesions.

Objective·To study early expression levels of serum heparin-binding protein(HBP)and its potential value in early alarming for prognosis and occurrence of sepsis in patients with severe bums.Methods·Retrospective analysis was performed on medical records of 52 severely burned patients admitted to the Department of Bum and Plastic,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine during January 2021 to May 2023.General data of patients on gender,age,total bum area,body mass index(BMI),and whether endotracheal intubation or incision was performed were collected.The level of HBP,serum procalcitonin(PCT),serum C-reactive protein(CRP)and the count of white blood cells within 48 h of admission were collected.The scores of acute physiological and chronic health assessment evaluation Ⅱ(APACHE Ⅱ)and sequential organ failure assessment(SOFA)within 48 h of admission were collected.Patients were divided into death group and survival group according to the status at discharge.According to whether sepsis occurred during hospitalization,the patients were divided into sepsis group and non-sepsis group.According to whether shock occurred,the septic patients were divided into sepsis without shock group and septic shock group.Risk factors for death,sepsis,and septic shock in severely burned patients were analyzed by using Logistic regression models.Receiver operator characteristic(ROC)curve analysis was established to study risk factors,which may alarm the occurrence of poor prognosis,sepsis,and septic shock.Results·Compared with the survival group,patients in the death group were older,and the difference was statistically significant(P=0.036).Differences in burn area and whether tracheal intubation or tracheotomy was performed were statistically significant in patients with or without sepsis(P=0.011,P=0.001).Compared with the survival group,the serum HBP levels were higher in the death group,and the difference was statistically significant(P=0.002).Compared with the non-sepsis group,patients in the sepsis group had higher levels of serum HBP,APACHE Ⅱ scores,and SOFA scores,and the differences were statistically significant(all P＜0.05).The differences in other indicators were not statistically significant.Compared with the sepsis without shock group,the septic shock group had higher HBP levels,with a statistically significant difference(P=0.008).Logistic regression analysis showed that HBP was an independent risk factor for death in patients with severe burns during hospitalization and also an independent risk factor for the occurrence of septic shock in patients with sepsis after severe burns.ROC curve analysis showed that the area under the curve(AUC)for HBP in predicting patient mortality during hospitalization was 0.798;when HBP ≥147.03 ng/mL,its sensitivity and specificity were 88.33％and 70.00％,respectively.The AUC for HBP in predicting the occurrence of septic shock in patients with sepsis after severe burns was 0.789;when HBP ≥147.03 ng/mL,its sensitivity and specificity were 90.00％and 63.20％,respectively.Conclusion·The level of serum HBP within 48 h of admission might be used as an early alarming index for prognosis in patients with severe burns and for the occurrence of septic shock in patients with sepsis following severe burns.

Objective·To systematically integrate relevant influencing factors of advanced cancer patients'engagement behavior in advance care planning(ACP).Methods·The systematic search of Chinese and English literature on factors influencing ACP engagement behavior in advanced cancer patients from inception to December 2022 in China National Knowledge Infrastructure(CNKI),Wanfang,China Biomedical Literature Database(Sinomed),PubMed,Cochrane Library,Embase,CINAHL,and PsycINFO was conducted.After the literature quality evaluation,content extraction and summary were conducted by two researchers,and the data of quantitative research and qualitative research were extracted and integrated respectively.The final influencing factors of ACP engagement behavior of advanced cancer patients were obtained.With the help of the theoretical domain,they were mapped to the capability,opportunity,motivation-behavior(COM-B)model step by step.Results·A total of 21 studies were included and 27 factors were summarized,including 9 theoretical domains.Mapping to the COM-B model included 9 capability factors(knowledge of ACP,education level,accurate knowledge of prognosis,knowledge of the time of disease diagnosis,prior experience,subjective life expectancy,age,cancer site,and disease symptom burden),13 opportunity factors(gender,marital status,race/ethnicity,religious belief,dependent children,family economic condition,place of living,housing type,family support,social support,doctor-patient relationship,acculturation,and whether or not to establish a hospice service center)and 5 motivational factors(ACP attitude,ACP belief,ACP motivation,anxiety and depression,and death attitude).Among them,doctor-patient relationship,religious belief,ACP attitude,educational level,marital status,family support,knowledge of ACP,accurate knowledge of prognosis,age,place of living,attitude toward death,prior experience,and race/ethnicity were more influential factors on ACP engagement behavior.Conclusion·Based on the COM-B model,the influencing factors of ACP engagement behavior in advanced cancer patients can be comprehensively summarized.Future studies can use the above factors as an entry point to design continuous,multifaceted,and comprehensive interventions based on the COM-B model to promote the practice of ACP engagement behavior in advanced cancer patients.

Diabetes mellitus type 2 might cause mild cognitive impairment in its advanced stages,potentially progressing to dementia.Diabetic cognitive impairment(DCI)stands as a chronic complication of diabetes mellitus,with its underlying pathogenesis still remaining elusive.Research has revealed that gut microbiota dysbiosis influenced the central nervous system through the"microbiota-gut-brain axis",thereby contributing to the progression of cognitive impairment.Therefore,the regulation of gut microbiota emerges as a promising approach to the prevention and treatment of DCI.This article comprehensively reviews the mechanisms through which gut microbiota influences DCI.Furthermore,it delves into experimental studies exploring targeted therapies for gut microbiota,including probiotics,fecal microbiota transplantation,dietary and nutrient interventions,as well as traditional Chinese medicine.These studies not only address diabetes-related cognitive impairment but also consider aspects such as glycolipid metabolism and inflammation.The insights gleaned from these studies provide valuable guidance for the clinical application of gut microbiota-targeted intervention in DCI.