Loss of protein homeostasis is a hallmark of cellular senescence, and ribosome pausing plays a crucial role in the collapse of proteostasis. However, our understanding of ribosome pausing in senescent cells remains limited. In this study, we utilized ribosome profiling and G-quadruplex RNA immunoprecipitation sequencing techniques to explore the impact of RNA G-quadruplex (rG4) on the translation efficiency in senescent cells. Our results revealed a reduction in the translation efficiency of rG4-rich genes in senescent cells and demonstrated that rG4 structures within coding sequence can impede translation both in vivo and in vitro. Moreover, we observed a significant increase in the abundance of rG4 structures in senescent cells, and the stabilization of the rG4 structures further exacerbated cellular senescence. Mechanistically, the RNA helicase DHX9 functions as a key regulator of rG4 abundance, and its reduced expression in senescent cells contributing to increased ribosome pausing. Additionally, we also observed an increased abundance of rG4, an imbalance in protein homeostasis, and reduced DHX9 expression in aged mice. In summary, our findings reveal a novel biological role for rG4 and DHX9 in the regulation of translation and proteostasis, which may have implications for delaying cellular senescence and the aging process.
G-Quadruplexes ; Cellular Senescence ; Ribosomes/genetics* ; Humans ; Animals ; Mice ; DEAD-box RNA Helicases/genetics* ; Protein Biosynthesis ; RNA/chemistry* ; Neoplasm Proteins

Objective:To evaluate the association between the preoperative aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio and postoperative delirium (POD) in patients undergoing total knee/hip arthroplasty.Methods:In this nested case-control study, medical records from patients, aged ≥55 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with Mini-Mental State Examination (MMSE) scale score >24 on preoperative day 1, scheduled for elective total knee/hip arthroplasty under combined spinal-epidural anesthesia at Qingdao Municipal Hospital between November 2021 and December 2023, were collected. POD was assessed using the Confusion Assessment Method. Patients were categorized into POD and non-POD groups based on the occurrence of POD. Logistic regression was employed to identify protective and risk factors for POD. The predictive performance of preoperative AST/ALT ratio alone and its combination with cerebrospinal fluid (CSF) biomarker concentrations for POD was evaluated using receiver operating characteristic curves, and the clinical utility of the prediction models was assessed using decision curve analysis. Mediation analysis was conducted to examine the mediation role of CSF biomarkers in the relationship between the preoperative AST/ALT ratio and POD. Restricted cubic splines were used to assess the nonlinear relationship between the preoperative AST/ALT ratio and POD.Results:A total of 460 patients were finally included, with 49 in POD group and 411 in non-POD group. After adjustment for multiple confounding factors including age, sex, educational level, MMSE score, history of hypertension, history of diabetes, history of smoking and history of alcohol use, multivariable logistic regression identified that the higher preoperative AST/ALT ratio, CSF total tau protein (t-tau) and phosphorylated tau protein (p-tau) concentrations were independent risk factors for POD, whereas lower CSF β-amyloid 42 (Aβ 42) concentration, Aβ 42/t-tau ratio and Aβ 42/p-tau ratio served as protective factors ( P<0.05). The area under the receiver operating characteristic curve of the preoperative AST/ALT ratio combined with CSF biomarkers in predicting POD was 0.939 ( P<0.001), demonstrating high clinical efficacy. After adjusting for age, educational level, sex, MMSE score, history of hypertension, history of diabetes, history of smoking, history of alcohol use and body mass index, restricted cubic splines revealed a nonlinear relationship between preoperative AST/ALT levels and the probability of POD, and the probability of POD increased with rising ratios when the preoperative AST/ALT ratio ranged from 1.09 to 1.40 ( Poverall < 0.05, Pnonlinear <0.05). After stratification by sex, the preoperative AST/ALT ratio demonstrated a linear relationship with the probability of POD ( Poverall <0.05, Pnonlinear>0.05). Mediation analysis indicated that the relationship between the preoperative AST/ALT ratio and POD was partially mediated by CSF p-tau concentration (proportion mediated 18.1%), CSF t-tau concentration (proportion mediated 12.0%), and the Aβ 42/t-tau ratio (proportion mediated 15.4%). Conclusions:A higher preoperative AST/ALT ratio is an independent risk factor for POD in total knee/hip arthroplasty patients. The concentrations of CSF t-tau and p-tau and Aβ 42/t-tau ratio have a mediating role in the relationship between the preoperative AST/ALT ratio and POD.

Objective To investigate the effects of Demodex infection on clinical symptoms,signs,and tear MMP-9 levels in patients with meibomian gland dysfunction(MGD).Methods A total of 680 patients with MGD were selected from our hospital,including 162 males and 518 females,with an average age of(45.05±15.41)years old.The patients were divided into two groups based on the presence of Demodex mite infestation:the Demodex positive group(340 cases)and the Demodex negative group(340 cases).All patients underwent evaluations using the OSDI questionnaire,SPEED questionnaire,eyelid margin alteration score,corneal fluorescein staining score,tear MMP-9 measurement,meibomian gland orifice score,meibomian gland excretion ability score,meibomian gland secretion score,meibomian gland loss score,tear film breakup time(BUT),and Schirmer I tear secretion test.The differences in these indicators between the two groups were compared.Results SPEED questionnaire score:Demodex positive group:(7.68±2.80),Demodex negative group:(6.28±1.99).There was a statistically significant difference between the two groups(t=2.582,P=0.012).Eyelid margin alteration score:Demodex positive group:(3.63±1.53),Demodex negative group:(2.85±0.77).A statistically significant difference was observed(t=2.861,P=0.006).Corneal fluorescein staining score:Demodex positive group:(2.25±1.86),Demodex negative group:(1.08±1.33).There was a statistically significant difference(t=3.247,P=0.002).Tear MMP-9 content:Demodex positive group:(30.76±43.14)ng/mL,Demodex negative group:(12.36±12.10)ng/mL.A statistically significant difference was found(t=2.598,P=0.013).No statistically significant differences were observed between the Demodex positive and negative groups in meibomian gland orifice score,meibomian gland excretion ability score,meibomian gland secretion score,meibomian gland loss score,BUT,tear secretion examination,and age comparison(P>0.05).Conclusions Demodex mite infestation in patients with MGD exhibits significant differ-ences across various clinical indicators,notably in SPEED questionnaire scores,eyelid margin alterations,corneal fluorescein staining,and tear MMP-9 levels.These changes are associated with mechanisms including inflammatory responses,cellular damage,and immune dysregulation.Demodex mite infestation may significantly influence the clinical progression of MGD by exacerbating inflammation and symptom severity,potentially playing a crucial role in disease development.

Objective:To evaluate the association between the preoperative aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio and postoperative delirium (POD) in patients undergoing total knee/hip arthroplasty.Methods:In this nested case-control study, medical records from patients, aged ≥55 yr, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with Mini-Mental State Examination (MMSE) scale score >24 on preoperative day 1, scheduled for elective total knee/hip arthroplasty under combined spinal-epidural anesthesia at Qingdao Municipal Hospital between November 2021 and December 2023, were collected. POD was assessed using the Confusion Assessment Method. Patients were categorized into POD and non-POD groups based on the occurrence of POD. Logistic regression was employed to identify protective and risk factors for POD. The predictive performance of preoperative AST/ALT ratio alone and its combination with cerebrospinal fluid (CSF) biomarker concentrations for POD was evaluated using receiver operating characteristic curves, and the clinical utility of the prediction models was assessed using decision curve analysis. Mediation analysis was conducted to examine the mediation role of CSF biomarkers in the relationship between the preoperative AST/ALT ratio and POD. Restricted cubic splines were used to assess the nonlinear relationship between the preoperative AST/ALT ratio and POD.Results:A total of 460 patients were finally included, with 49 in POD group and 411 in non-POD group. After adjustment for multiple confounding factors including age, sex, educational level, MMSE score, history of hypertension, history of diabetes, history of smoking and history of alcohol use, multivariable logistic regression identified that the higher preoperative AST/ALT ratio, CSF total tau protein (t-tau) and phosphorylated tau protein (p-tau) concentrations were independent risk factors for POD, whereas lower CSF β-amyloid 42 (Aβ 42) concentration, Aβ 42/t-tau ratio and Aβ 42/p-tau ratio served as protective factors ( P<0.05). The area under the receiver operating characteristic curve of the preoperative AST/ALT ratio combined with CSF biomarkers in predicting POD was 0.939 ( P<0.001), demonstrating high clinical efficacy. After adjusting for age, educational level, sex, MMSE score, history of hypertension, history of diabetes, history of smoking, history of alcohol use and body mass index, restricted cubic splines revealed a nonlinear relationship between preoperative AST/ALT levels and the probability of POD, and the probability of POD increased with rising ratios when the preoperative AST/ALT ratio ranged from 1.09 to 1.40 ( Poverall < 0.05, Pnonlinear <0.05). After stratification by sex, the preoperative AST/ALT ratio demonstrated a linear relationship with the probability of POD ( Poverall <0.05, Pnonlinear>0.05). Mediation analysis indicated that the relationship between the preoperative AST/ALT ratio and POD was partially mediated by CSF p-tau concentration (proportion mediated 18.1%), CSF t-tau concentration (proportion mediated 12.0%), and the Aβ 42/t-tau ratio (proportion mediated 15.4%). Conclusions:A higher preoperative AST/ALT ratio is an independent risk factor for POD in total knee/hip arthroplasty patients. The concentrations of CSF t-tau and p-tau and Aβ 42/t-tau ratio have a mediating role in the relationship between the preoperative AST/ALT ratio and POD.

Objective To investigate the effects of Demodex infection on clinical symptoms,signs,and tear MMP-9 levels in patients with meibomian gland dysfunction(MGD).Methods A total of 680 patients with MGD were selected from our hospital,including 162 males and 518 females,with an average age of(45.05±15.41)years old.The patients were divided into two groups based on the presence of Demodex mite infestation:the Demodex positive group(340 cases)and the Demodex negative group(340 cases).All patients underwent evaluations using the OSDI questionnaire,SPEED questionnaire,eyelid margin alteration score,corneal fluorescein staining score,tear MMP-9 measurement,meibomian gland orifice score,meibomian gland excretion ability score,meibomian gland secretion score,meibomian gland loss score,tear film breakup time(BUT),and Schirmer I tear secretion test.The differences in these indicators between the two groups were compared.Results SPEED questionnaire score:Demodex positive group:(7.68±2.80),Demodex negative group:(6.28±1.99).There was a statistically significant difference between the two groups(t=2.582,P=0.012).Eyelid margin alteration score:Demodex positive group:(3.63±1.53),Demodex negative group:(2.85±0.77).A statistically significant difference was observed(t=2.861,P=0.006).Corneal fluorescein staining score:Demodex positive group:(2.25±1.86),Demodex negative group:(1.08±1.33).There was a statistically significant difference(t=3.247,P=0.002).Tear MMP-9 content:Demodex positive group:(30.76±43.14)ng/mL,Demodex negative group:(12.36±12.10)ng/mL.A statistically significant difference was found(t=2.598,P=0.013).No statistically significant differences were observed between the Demodex positive and negative groups in meibomian gland orifice score,meibomian gland excretion ability score,meibomian gland secretion score,meibomian gland loss score,BUT,tear secretion examination,and age comparison(P>0.05).Conclusions Demodex mite infestation in patients with MGD exhibits significant differ-ences across various clinical indicators,notably in SPEED questionnaire scores,eyelid margin alterations,corneal fluorescein staining,and tear MMP-9 levels.These changes are associated with mechanisms including inflammatory responses,cellular damage,and immune dysregulation.Demodex mite infestation may significantly influence the clinical progression of MGD by exacerbating inflammation and symptom severity,potentially playing a crucial role in disease development.

This article reported a case of hypertrophic pyloric stenosis after neonatal esophageal atresia repair. The mother of the child did not have regular prenatal care. The child was born at a gestational age of 40 weeks and 2 days of gestation, with polyhydramnios at birth, and was diagnosed with esophageal atresia and cleft palate after birth and underwent thoracoscopic esophageal-esophageal end-to-end anastomosis and esophageal-tracheal fistula ligation and was given nasogastric feeding after surgery. At four months of age, the child vomited a lot of coffee-like material after nasogastric feeding, and the ultrasonographic and upper gastroenterography findings suggested hypertrophic pyloric stenosis, which was treated surgically with good results. This case suggests that hypertrophic pyloric stenosis should be considered in children with unexplained non-bilious vomiting/feeding difficulties after esophageal atresia repair. After definitive diagnosis, laparoscopic pyloromyotomy is feasible.

Objective:To discuss the effect of microRNA(miR)-30c-5p on the proliferation,migration,and invasion of the human prostate cancer cells(LNCap),and to clarify its possible mechanism.Methods:The LNCap cells were divided into LNCap group(without plasmid transfection),miR-30c-5p mimic group(transfected with miR-30c-5p mimic),mimic NC group(transfected with miR-30c-5p mimic NC),sh-DNA damage inducible transcript 4(DDIT4)group(transfected with sh-DDIT4),sh-NC group(transfected with sh-DDIT4 NC),miR-30c-5p mimic+pc-DNA3.1-NC group(co-transfected with miR-30c-5p mimic and pc-DNA3.1 empty vector),and miR-30c-5p mimic+pc-DNA3.1-DDIT4 group(co-transfected with miR-30c-5p mimic and pc-DNA3.1-DDIT4 over-expression plasmid).The RWPE-1 cells were cultured normally.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-30c-5p and DDIT4 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of DDIT4 protein in the cells in various groups;CCK-8 method was used to detect the proliferation rates of the LNCap cells in various groups;Transwell assay was used to detect the numbers of the invasion LNCap cells in various groups;Scratch assay was used to detect the scratch healing rates of LNCap cells in various groups;dual-luciferase reporter assay was used to detect the targeting relationship between miR-30c-5p and DDIT4.In the in vivo tumor formation experiment,18 male BALB/c nude mice were divided randomly into blank group,agomiR-NC group(transfected with agomiR-30c-5p NC),and agomiR-30c-5p group(transfected with agomiR-30c-5p);there were six mice in each group.The mice in agomiR-NC group and agomiR-30c-5p group were subcutaneously injected with LNCap cells,while the mice in blank group were given an equal volume of physiological saline.The volumes of tumor of the mice in various groups were detected.HE staining was used to observe the morphology of prostate cancer tissue the mice of in various groups;RT-qPCR method and immunofluorescence staining were used to detect the expression levels of miR-30c-5p and DDIT4 mRNA and the fluorescence intensities of DDIT4 protein in prostate cancer tissue of the mice in various groups.Results:The In vitro prostate cancer cell experiment results showed that compared with RWPE-1 cells,the expression level of miR-30c-5p in the prostate cancer LNCap cells was decreased(P<0.01),and the expression levels of DDIT4 mRNA and protein were increased(P<0.05 or P<0.01).After 48 of transfection,compared with LNCap group and mimic NC group,the expression level of miR-30c-5p in the LNCap cells in miR-30c-5p mimic group was increased(P<0.01).Compared with LNCap group and sh-NC group,the expression level of DDIT4 mRNA in the LNCap cells in sh-DDIT4 group was decreased(P<0.01).Compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of miR-30c-5p in The LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 mRNA in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 protein in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.05).The CCK-8 method results showed that compared with LNCap group and mimic NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the proliferation rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The Transwell assay results showed that compared with LNCap group and mimic NC group,the number of the invasion LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the number of invasion LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the number of the invasion LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The scratch assay results showed that compared with LNCap group and mimic NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the scratch healing rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The dual-luciferase reporter assay results showed that compared with the LNCap cells co-transfected with WT-DDIT4 and mimic NC,the luciferase activity of the LNCap cells co-transfected with WT-DDIT4 and miR-30c-5p mimic was decreased(P<0.01).The in vivo nude mouse tumor formation experiment results showed that on the 3 rd,6 th,9 th,12 th,and 15th days after cell injection,compared with blank group and agomiR-NC group,the tumor volumes of the nude mice in agomiR-30c-5p group were decreased(P<0.05).The HE staining results showed that in prostate cancer tissue of the mice in blank group and agomiR-NC group,the cell nuclei were enlarged,and nucleoli were prominent and deformed.In the mice in agomiR-30c-5p group,some regions of prostate cancer tissues results showed neatly arranged cells with normally shaped nuclei.The RT-qPCR and immunofluorescence staining showed that compared with agomiR-NC group,the expression level of miR-30c-5p in prostate cancer tissue of the mice in agomiR-30c-5p group was increased(P<0.01).Compared with blank group and agomiR-NC group,the expression level of DDIT4 mRNA in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).DDIT4 protein was mainly expressed in the cytoplasm.Compared with blank group and agomiR-NC group,the fluorescence intensity of DDIT4 protein in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).Conclusion:The expression level of miR-30c-5p in the prostate cancer LNCap cells is decreased,and it inhibits the proliferation,migration,and invasion of the prostate cancer cells by targeting downregulation of DDIT4,thereby participating in the occurrence and development of prostate cancer.

Sarcopenia and physical deconditioning are common complications in patients with liver cancer,which are frequently caused by insufficient physical activity and poor nutritional status,resulting in physical frailty and a significant impact on the patient's physical fitness.Notably,sarcopenia,frailty,and poor cardiopulmonary endurance have all been linked to higher mortality rates among patients with liver cancer.Exercise intervention significantly improves various health parameters in liver cancer patients,including metabolic syndrome,muscle wasting,cardiorespiratory endurance,health-related quality of life,and reduction in hepatic venous pressure gradient.However,the link between physical exercise and liver cancer is commonly overlooked.In this article,we will examine the impact of exercise on liver cancer and present the most recent evidence on the best types of exercise for various stages of liver cancer.This article also summarizes and discusses the molecular mechanisms that control metabolism and systemic immune function in tumors.In brief,physical exercise should be considered an important intervention in the prevention and treatment of liver cancer and its complications.

Background Alzheimer's disease(AD)is the most common type of dementia.There has been an increasing trend in incidence rate.The drug therapy has been proven ineffective in improving the cognitive function of AD patients or delaying the disease progression,leading to perceived low adherence with treatment and low efficacy.Previous basic research shows that blue light irradiation is helpful to improve the cognitive function of AD model rats,but there is no corresponding clinical study on the improvement effect of blue light irradiation on cognitive function of AD patients.Objective To explore the efficacy of blue light irradiation in improving the cognitive function of AD patients,so as to provide references for cognitive stabilization or improvement of AD patients.Methods A sample of 155 patients who were hospitalized in Geriatric Department of Shenzhen Kangning Hospital from June 2019 to December 2023 and met the International Classification of Diseases,tenth edition(ICD-10)diagnostic criteria for AD were retrospectively selected.Among them,80 cases who received blue light irradiation combined with memantine therapy and conventional psychotherapy were the study group,and 75 cases who only received memantine therapy and conventional psychotherapy were the control group.Blue light irradiation therapy lasted for 4 weeks,5 times per week,with each session lasting for 30 min.Then Mini-Mental State Examination(MMSE)and Activity of Daily Living Scale(ADL)were administered at the baseline and after treatment.Results After treatment,study group reported an increase in MMSE total score,orientation,immediate recall,attention/calculation,delayed recall and language dimension scores(Z=-6.931,-5.773,-4.123,-3.649,-3.508,-4.733,P<0.05),and a decrease in ADL total score(Z=-7.020,P<0.05)compared to baseline data.Analysis indicated that study group scored higher on MMSE total scale,orientation,immediate recall and language dimensions(Z=-2.784,-4.621,-2.483,-3.463,P<0.05),and lower on ADL total score(Z=-3.704,P<0.05)than those of control group after treatment.Conclusion Blue light irradiation for AD patients may improve their cognitive function and activities of daily living.