Objective To observe the effect of a heavy load exercise on the ultrastructure,function and fission of skeletal muscle mitochondria in rats,and to analyze the changes of the phosphorylation expression of mitochondrial fission protein and upstream kinase at different times postexercise,and to explore the effect of acute heavy load exercise on mitochondrial fission in skeletal muscle of rats and its possible mechanism.Methods Forty-eight adult male Sprague-Dawley rats were randomly divided in-to a quiet control group(C,n=8)and an exercise group(E,n=40).Rats in the E group exercised on a treadmill down a 16° decline at 16 m/min for 90 min and were further divided into 0 h,12 h,24 h,48 h,and 72 h postexercise subgroups.Soleus muscle was isolated and mitochondria were ex-tracted at the corresponding time points after exercise.The ultrastructure of mitochondria in the soleus muscle was observed using transmission electron microscopy,and mitochondrial quantity and morphomet-ric analysis were conducted.Moreover,the colocalization and quantity of dynamin-related protein 1(Drp1)and cytochrome C oxidease subunit Ⅳ(COXⅣ)in the soleus muscle were detected using im-munofluorescence double-labeling techniques.Meanwhile,protein levels of soleus musclep-Drp1Ser616,p-Drp1Ser637,p-extracellular regulatory protein kinaseThr202/Tyr204(p-ERKThr202/Tyr204),p-protein kinaseAThr197(p-PKAThr197),and mitochondrial NADH of ubiquinone oxidoreductase subunit B8(NDUFB8)and ubiqui-nol-cytochrome C reductase core protein 2(UQCRC2)were determined by using Western blotting.An-other twenty-four rats were randomly divided into a DMSO group(CD),a U0126 group(CU),an Ex-ercise+DMSO group(ED),and an Exercise+U0126 group(EU).Six mice in each group were giv-en a single intra-bitoneal injection of DMSO or ERK inhibitor U0126 20 min before acute downhill running.Then,their phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 in soleus muscle were de-tected by Western blotting.Results(1)From 0 h to 48 h after exercise,the soleus muscle mitochon-dria showed swelling,rounding,and uneven distribution of mitochondria,among which the degree of mitochondrial damage was the most serious at 12 h and 24 h after exercise.Moreover,the protein ex-pression of NDUFB8 and UQCRC2 in the mitochondria fractions from soleus muscle was significantly lower at 12 h post-exercise(P<0.05).(2)The co-localization of Drp1 and COXⅣ in the skeletal muscle increased significantly at 12 h to 24 h after a heavy load exercise compared with group C and group E0(P<0.01).Moreover,the mitochondrial area,circumference,aspect ratio and Ferret diameter in the skeletal muscle were significantly lower at 12 h to 24 h postexercise(P<0.05).Meanwhile,the number of mitochondria was significantly higher at 24 h after exercise(P<0.01).(3)The phosphoryla-tion of ERKThr202/Tyr204,PKAThr197 and Drp1Ser616 was significantly higher at 24 h after exercise(P<0.01),while that of Drp1Ser637 was significantly lower at 48 h and 72 h post-exercise(P<0.01).However,the phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 were significantly down-regulated by U0126 treatment before exercise.Conclusion A session of heavy load exercise caused mitochondrial structure and function damage and induced mitochondrial fission in the skeletal muscle,and then to maintain the homeostasis of skeletal muscle cells by cleaving damaged mitochondria.The mechanism of promot-ing skeletal muscle repair may be related to the positive and negative regulation of Drp1 activity by the phosphorylation of Drp1Ser616 and Drp1Ser637,respectively.Among them,the activation of ERKThr202/Tyr204 mediates the phosphorylation activation of Drp1Ser616,but PKAThr197 is not an upstream kinase that medi-ates the inactivation of Drp1Ser637 phosphorylation.

Objective To explore the mechanism by which Erianin(ER)inhibits the high glucose(HG)-induced ep-ithelial-mesenchymal transition of ARPE-19 cells through the HIPPO/YeS-associated protein(YAP)signaling pathway.Methods The CCK-8 assay was used to determine the optimal glucose concentration(which was 50 mmol·L-1)for in-ducing the proliferation of ARPE-19 cells under HG conditions.The cytotoxic effects of different concentrations of ER(in-cluding 10,20,and 40 nmol·L-1)on ARPE-19 cells were evaluated.ARPE-19 cells were divided into the control group(cultured in a normal medium for 48 h),the HG group(cultured in a medium supplemented with 50 mumol·L-1 glucose for 48 h),and the HG `+low/medium/high ER groups(cultured in a medium with 50 mmol·L-1 glucose for 48 h after pre-treatment with 10,20,and 40 nmol·L-1 ER for 1 h,respectively).The CCK-8 assay was performed to assess the prolifera-tion rate of ARPE-19 cells.The scratch assay was used to evaluate cell migration activity and calculate the wound healing percentage.Immunofluorescence was employed to detect the intracellular expression of Vimentin in ARPE-19 cells.West-ern blot analysis was used to determine the intracellular expression levels of α-smooth muscle actin(α-SMA),neural cad-herin(N-Cadherin),epithelial cadherin(E-Cadherin),mammalian sterile 20-like kinase 1(MST1),large tumor suppressor kinase 1(LATS1),YAP,and phosphorylated YAP(P-YAP)in ARPE-19 cells.Results Compared with those in the con-trol group,the proliferation rate,the wound healing percentage,and the immunofluorescence intensity of Vimentin proteins in ARPE-19 cells in the HG group were significantly increased(all P＜0.01).The relative expression levels of α-SMA,N-Cadherin,and YAP proteins were significantly elevated while the relative expression levels of E-cadherin,P-YAP,MST1,and LATS1 proteins were significantly decreased in the HG group than those in the control group(all P＜0.01).ARPE-19 cells in the HG+low/medium/high ER groups exhibited a lower cell proliferation rate,a lower wound healing percentage,and weakened Vimentin fluorescence intensity than those in the HG group(all P＜0.05).The relative expression levels of YAP,α-SMA,and N-cadherin proteins were significantly reduced while the relative expression levels of E-cadherin and P-YAP were significantly increased in the HG+low/medium/high ER groups,compared with those in the HG group(all P＜0.05).The relative expression levels of MST1 and LATS1 proteins in the HG+medium/high ER groups were significantly higher than those in the HG group(both P＜0.05).Conclusion ER can inhibit the HG-induced epithelial-mesenchymal transition of ARPE-19 cells,which may be related to the regulation of the Hippo/YAP signaling pathway.

Objective:To investigate the role of microRNA-709(miR-709)in erythroid development and its correlation with multiple hematological diseases.Methods:The expression of miR-709 in multiple tissues of mice was detected by qRT-PCR;The expression of miR-709 and other miRNAs in day-14.5 fetal liver cells from mouse embryos was detected by transcriptome microarray analysis;The expression of miR-709 in nucleated red blood cells derived from bone marrow and spleen,and in peripheral blood erythrocytes of mice was detected by magnetic bead sorting combined with qRT-PCR;The expression of miR-709 during erythroid differentiation of murine erythroleukemia(MEL)cells was analyzed by cell culture and qRT-PCR;The expression of miR-709 during erythroid lineage differentiation of mouse erythroid precursor cells derived from fetal livers was analyzed by magnetic bead sorting,flow cytometry,cell culture and qRT-PCR;The expression of miR-709 in peripheral blood of patients with different hematological diseases was measured by qRT-PCR.Results:Among the various tissues examined in mice,miR-709 exhibited the highest expression in peripheral blood,followed by high expression levels in muscle,bone marrow,and liver;In day-14.5 fetal liver cells from mouse embryos,miR-709 was highly expressed,significantly surpassing miR-451,which was most highly expressed in mature red blood cells;In nucleated red blood cells derived from mouse bone marrow and spleen,the expression of miR-709 was higher than that of miR-451,whereas the opposite pattern was observed in peripheral blood;During the differentiation of erythroid precursor cells derived from mouse embryonic liver,the expression level of miR-709 first increased and then decreased;During the differentiation of MEL cells,the expression of miR-709 gradually increased;Compared with the healthy controls,patients with myelodysplastic syndrome(MDS),α-thalassemia and β-thalassemia expressed lower levels of miR-709 in peripheral blood;while the expression of miR-709 in patients with infectious hemolytic anemia(IHA)was higher than that in healthy controls.Conclusion:miR-709 is highly expressed in the early stage of erythropoiesis and exhibits dynamic changes during erythroid development,potentially playing an important role.It is also differentially expressed in different hematological diseases,which is expected to serve as a promising biomarker and therapeutic target in the clinical diagnosis and treatment of hematological diseases.

BACKGROUND:The prognosis of patients undergoing percutaneous transforaminal endoscopic discectomy is poor,which is related to the lack of functional physical exercise.Conventional rehabilitation methods have limited efficacy,mainly because the rehabilitation methods are relatively single and lack specificity.Therefore,it is necessary to improve the overall functional physical exercise method.OBJECTIVE:To explore the effect of overall functional physical exercise on lumbar biomechanics in patients with lumbar disc herniation who underwent percutaneous transforaminal endoscopic discectomy.METHODS:120 patients who met the percutaneous intervertebral foramen discectomy indication and underwent percutaneous intervertebral foramen discectomy operation in Henan Provincial People's Hospital from April 2021 to September 2022 were selected as study subjects.They were randomly divided into traditional rehabilitation group and overall functional physical exercise group.Patients in the two groups received 8 weeks of traditional rehabilitation and overall functional physical exercise respectively.Before the exercise intervention and after 8 weeks of intervention,the lsoMed2000 isokinetic muscle strength testing system was used to conduct isokinetic muscle strength testing.The Oswestry Disability Index was used to evaluate lumbar spine function.The visual analog scale was used to evaluate the pain level.The anxiety self-rating scale and depression self-report scale were used to assess symptoms of anxiety and depression.After 8 weeks of exercise intervention,the modified Macnab efficacy evaluation standard was used to evaluate the efficacy.RESULTS AND CONCLUSION:(1)Compared with the traditional rehabilitation group,the peak torque and average power of flexion and extensor in the whole functional physical exercise group increased,the peak torque ratio of flexion and extension decreased,Oswestry disability index score,visual analog scale score,self-rating anxiety scale score and self-rating depression scale score decreased,and the excellent and good rate increased 8 weeks after treatment(P＜0.001).(2)It is concluded that compared with the traditional rehabilitation program,overall functional physical exercise after percutaneous intervertebral foramen discectomy in patients with lumbar disc herniation can effectively enhance lumbar biomechanics and function,relieve pain,reduce postoperative bad mood,and improve prognosis,which has high clinical value.

Objective:This study aims to investigate the predictive performance of serum levels of white blood cell count(WBC),D-dimer(D-D)and N-terminal pro-brain natriuretic peptide(NT-proBNP)for major adverse cardiovascular events(MACE)in elderly patients with acute ST-segment elevation myocardial infarction(ASTE-MI).Methods:A total of 70 elderly patients with ASTEMI treated in Bozhou People's Hospital between April 2020 and May 2023 were prospectively selected as observation group.Incidence of MACE during 1-year follow-up were recorded,another 50 patients with unstable angina pectoris treated in our hospital simultaneously were selected as control group.Serum levels of WBC,D-D and NT-proBNP were compared among above groups.The receiver operating characteristic(ROC)curve was used to evaluate the predictive value of serum WBC,D-D and NT-proBNP for MACE in elderly patients with ASTEMI.A nomogram was established,and calibration curve and deci-sion curve analysis(DCA)were used to evaluate the performance of model.Results:A total of 40 cases(59.70％)experienced MACE during one-year follow-up.Compared to those in control group,patients in observation group had significant higher serum WBC[(11.43±1.98)×109/Lvs.(6.30±1.99)× 109/L],D-D[(0.91±0.20)mg/L vs.(0.47±0.18)mg/L]andNT-proBNP[(192.31±63.19)pg/ml vs.(114.05±22.79)pg/ml](P＜0.001 all).Compared to participants without MACE,those with MACE had significantly higher serum WBC[(13.33±1.90)× 109/L vs.(10.27±0.98)× 109/L],D-D[(1.11±0.25)mg/L vs.(0.87±0.21)mg/L]and NT-proBNP[(238.73±50.22)pg/ml vs.(150.70±39.16)pg/ml](P＜0.001 all).ROC analysis showed that the ar-ea under the curve(AUC)of the combined detection of serum WBC(AUC=0.791,95％CI 0.677～0.879),D-D(AUC=0.767,95％CI 0.650～0.859)and NT-proBNP(AUC=0.733,95％CI 0.614～0.832)was 0.916(95％CI 0.825～0.969),which was significantly higher than those of single detections(Z=2.386,4.953,3.190,P=0.017,0.004,＜0.001).The total score of the nomogram model constructed based on the levels of WBC,D-D and NT-proBNP ranged from 70 to 126 points.The predicted incidence was basically consistent with the actual in-cidence.For the internal verification of the model,the AUC of ROC curve of the training set and the validation set was 0.863 and 0.926 respectively.The DCA curve was located above the critical curve,indicating that the model had a net benefit and good clinical effectiveness.Conclusion:Serum WBC,D-D and NT-proBNP significantly el-evated in elderly patients with ASTEMI.The combined detection of serum WBC,D-D and NT-proBNP levels has good predictive value for MACE in these patients.

Objective To observe the effect of a heavy load exercise on the ultrastructure,function and fission of skeletal muscle mitochondria in rats,and to analyze the changes of the phosphorylation expression of mitochondrial fission protein and upstream kinase at different times postexercise,and to explore the effect of acute heavy load exercise on mitochondrial fission in skeletal muscle of rats and its possible mechanism.Methods Forty-eight adult male Sprague-Dawley rats were randomly divided in-to a quiet control group(C,n=8)and an exercise group(E,n=40).Rats in the E group exercised on a treadmill down a 16° decline at 16 m/min for 90 min and were further divided into 0 h,12 h,24 h,48 h,and 72 h postexercise subgroups.Soleus muscle was isolated and mitochondria were ex-tracted at the corresponding time points after exercise.The ultrastructure of mitochondria in the soleus muscle was observed using transmission electron microscopy,and mitochondrial quantity and morphomet-ric analysis were conducted.Moreover,the colocalization and quantity of dynamin-related protein 1(Drp1)and cytochrome C oxidease subunit Ⅳ(COXⅣ)in the soleus muscle were detected using im-munofluorescence double-labeling techniques.Meanwhile,protein levels of soleus musclep-Drp1Ser616,p-Drp1Ser637,p-extracellular regulatory protein kinaseThr202/Tyr204(p-ERKThr202/Tyr204),p-protein kinaseAThr197(p-PKAThr197),and mitochondrial NADH of ubiquinone oxidoreductase subunit B8(NDUFB8)and ubiqui-nol-cytochrome C reductase core protein 2(UQCRC2)were determined by using Western blotting.An-other twenty-four rats were randomly divided into a DMSO group(CD),a U0126 group(CU),an Ex-ercise+DMSO group(ED),and an Exercise+U0126 group(EU).Six mice in each group were giv-en a single intra-bitoneal injection of DMSO or ERK inhibitor U0126 20 min before acute downhill running.Then,their phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 in soleus muscle were de-tected by Western blotting.Results(1)From 0 h to 48 h after exercise,the soleus muscle mitochon-dria showed swelling,rounding,and uneven distribution of mitochondria,among which the degree of mitochondrial damage was the most serious at 12 h and 24 h after exercise.Moreover,the protein ex-pression of NDUFB8 and UQCRC2 in the mitochondria fractions from soleus muscle was significantly lower at 12 h post-exercise(P<0.05).(2)The co-localization of Drp1 and COXⅣ in the skeletal muscle increased significantly at 12 h to 24 h after a heavy load exercise compared with group C and group E0(P<0.01).Moreover,the mitochondrial area,circumference,aspect ratio and Ferret diameter in the skeletal muscle were significantly lower at 12 h to 24 h postexercise(P<0.05).Meanwhile,the number of mitochondria was significantly higher at 24 h after exercise(P<0.01).(3)The phosphoryla-tion of ERKThr202/Tyr204,PKAThr197 and Drp1Ser616 was significantly higher at 24 h after exercise(P<0.01),while that of Drp1Ser637 was significantly lower at 48 h and 72 h post-exercise(P<0.01).However,the phosphorylated expressions of ERKThr202/Tyr204 and Drp1Ser616 were significantly down-regulated by U0126 treatment before exercise.Conclusion A session of heavy load exercise caused mitochondrial structure and function damage and induced mitochondrial fission in the skeletal muscle,and then to maintain the homeostasis of skeletal muscle cells by cleaving damaged mitochondria.The mechanism of promot-ing skeletal muscle repair may be related to the positive and negative regulation of Drp1 activity by the phosphorylation of Drp1Ser616 and Drp1Ser637,respectively.Among them,the activation of ERKThr202/Tyr204 mediates the phosphorylation activation of Drp1Ser616,but PKAThr197 is not an upstream kinase that medi-ates the inactivation of Drp1Ser637 phosphorylation.

Objective To explore the feasibility and corresponding implementation methods of constructing a sample resource bank based on individual-level data of completed clinical trials and using it to construct external controls for drug/device clinical trials.Methods Taking the pre-marketing clinical trial of transcatheter active valve replacement(TAVR)for the treatment of aortic valve stenosis as an example,the individual-level databases of multiple trials were standardized to form a sample bank.The original data of any trial in the sample bank were selected as the experimental group,and the remaining samples were selected as the control group.The potential confounding was handled by using the propensity score matching and stratification methods to clarify the process of constructing external controls based on the sample bank of individual-level data of clinical trials.Results This study included individual-level data of single-group trials of 4 TAVR devices,with a total of 569 subjects(59.2%male).The number of subjects in Trials 1 to 4 was 120,120,163,and 166,respectively.Propensity score matching enabled the matching of 113,117,125,and 147 subjects with comparable or similar characteristics from individual-level data from other trials,respectively,demonstrating a high matching success rate.The PS score distribution plot after stratification showed that the proportions of subjects in the experimental and control groups in strata 1 to 5 in scheme 1 were 4/103,11/103,22/92,32/87,and 51/64,respectively.For all constructed external controlled trials,a certain number of control samples with similar baseline characteristics to the experimental groups were distributed within each propensity score stratum.The results of the simulation test also reflected the potential differences between different devices in the 12-month all-cause mortality rate.Conclusions The sample bank constructed with individual-level data from clinical trials,as a high-quality data source,can serve as a source of external control for single-arm trials in the same field,and as a useful supplement to the external control scenario of real-world evidence to support drug and device development.At the same time,targeted research on research methods and bias control measures in related fields is also needed.

Chronic cerebral hypoperfusion leads to white matter injury (WMI), which plays a significant role in contributing to vascular cognitive impairment. While 13-docosenamide is a type of fatty acid amide, it remains unclear whether it has therapeutic effects on chronic cerebral hypoperfusion. In this study, we conducted bilateral common carotid artery stenosis (BCAS) surgery to simulate chronic cerebral hypoperfusion-induced WMI and cognitive impairment. Our findings showed that 13-docosenamide alleviates WMI and cognitive impairment in BCAS mice. Mechanistically, 13-docosenamide specifically binds to cannabinoid receptor 1 (CNR1) in oligodendrocyte precursor cells (OPCs). This interaction results in an upregulation of ubiquitin-specific peptidase 33 (USP33)-mediated CNR1 deubiquitination, subsequently increasing CNR1 protein expression, activating the phosphorylation of the AKT/mTOR pathway, and promoting the differentiation of OPCs. In conclusion, our study suggests that 13-docosenamide can ameliorate chronic cerebral hypoperfusion-induced WMI and cognitive impairment by enhancing OPC differentiation and could serve as a potential therapeutic drug.
Animals ; Oligodendrocyte Precursor Cells/metabolism* ; Mice ; Cell Differentiation/drug effects* ; Male ; Receptor, Cannabinoid, CB1/metabolism* ; Mice, Inbred C57BL ; Ubiquitin Thiolesterase/metabolism* ; Ubiquitination/drug effects* ; Carotid Stenosis/complications* ; Cognitive Dysfunction/drug therapy*

Objective To analyze the relationship between frailty and acute kidney injury(AKI)after hip fracture surgery in the elderly.Methods A total of 405 elderly patients who underwent hip fracture surgery in Jieyang People's Hospital from August 2021 to January 2023 were retrospectively analysed.According to the modified frailty index(mFI),they were divided into frail group(mFI≥0.27,n=112)and non-frail group(mFI<0.27,n=293).Postoperative AKI was defined according to the Kidney Disease:Improving Global Outcomes(KDIGO)criteria.After 1:1 propensity score matching(PSM),100 cases in each group were successfully matched.Univariable and multivariable logistic regression models,propensity score adjustment,PSM,inverse probability of treatment weighting(IPTW),standardized mortality ratio weighting(SMRW),pairwise algorithm(PA)weighting,and overlap weighting(OW)methods were used to analyze the relationship between frailty and postoperative AKI.Stratified analyses were performed according to age(≥80 or<80 years),gender,whether angiotensin-converting enzyme inhibitors(ACEI)/angiotensin Ⅱreceptor antagonists(ARB)were used,and whether intraoperative hypotension occurred.Results After PSM,there were no significant differences between the two groups in age,sex,surgical type,ACEI/ARB,blood urea nitrogen,serum creatinine,intraoperative blood loss,and intraoperative hypotension[standardized mean difference(SMD)<0.1].In both the original cohort and the matched cohort,the incidence of AKI was higher in frail group than in non-frail group(25.9%vs.8.9%,P<0.001;28.0%vs.11.0%,P=0.002).Analysis of the risk of postoperative AKI in elderly hip fracture patients in frail group found that compared with the non-frail group,in the univariate logistic regression model the odds ratio(OR)and a 95%confidence interval(CI)for the frail group was 3.59(2.00-6.43),P<0.001,and in the multivariable logistic regression model,the OR(95%CI)for frail group was 3.04(1.55-5.95),P=0.001.After adjustment for propensity score,the OR(95%CI)for frail group was 2.85(1.52-5.34),P=0.001,and the OR(95%CI)for frail group after PSM was 3.15(1.47-6.75),P=0.003.After IPTW,SMRW,PA weighting,and OW,the OR(95%CI)for frail group were 2.48(1.37-4.50),2.43(1.41-4.19),2.63(1.25-5.54),and 2.69(1.07-6.78),respectively,with P<0.05.The interaction tests were not statistically significant for age,sex,use of ACEI/ARB,and intraoperative hypotension(P>0.05).Conclusion In elderly patients with hip fractures,preoperative frailty may be a risk factor for postoperative AKI.

Objective To explore the effect of prominent cortical vein(PCV)sign on the clinical prognosis of patients undergoing endovascular recanalization for acute anterior circulation cerebral infarction.Methods A total of 102 patients with acute anterior circulation large-vessel occlusion in Zhongshan Municipal Hospital of Tradiontional Chinese Medicine from January 1,2020 to December 21,2022,who received endovascular recanalization,were enrolled in this study.The clinical data and radiographic materials were collected.According to whether PCV sign was present or not,the patients were divided into PCV group and non-PCV group.The clinical data were compared between the two groups.Logistic regression analysis was used to analyze the correlation between the postoperative 90-day mRS score and the occurrence of PCV.Results Compared with non-PCV group,in PCV group the incidence of atrial fibrillation was even higher,and the good rate of collateral compensation was even lower,and the postoperative(7±2)-day NIHSS score was even lower;the differences in above indexes between the two groups were statistically significant(all P＜0.05).Ninety days after treatment,the proportion of patients having modified Rankin Scale(mRS)score ≤2 points in the non-PCV group was 76.3％,which was higher than 58.6％in the PCV group,but the difference was not statistically significant(P=0.074).Logistic regression analysis indicated that a positive correlation existed between the presence of PCV sign and atrial fibrillation(OR=9.679,95％CI=1.264-74.364)and a negative correlation existed between the presence of PCV sign and good collateral compensation(OR=0.015,95％CI=0.033-0.331).No relationship existed between the postoperative 90-day good prognosis and the presence of preoperative PCV sign(OR=2.078,95％CI=0.680-6.348),but patient's age,preoperative intravenous thrombolysis were negatively correlated with the preoperative Alberta Stroke Program Early CT(ASPECT)score(P＜0.005).Conclusion In patients with anterior circulation acute ischemic stroke(AIS),the occurrence of PCV sign is associated with the impaired collateral compensation and atrial fibrillation,but its impact on the clinical outcomes of patients after receiving reperfusion therapy remains uncertain.PCV sign may be used as a potential evaluation indicator for inadequate cerebrovascular collateral compensation.