Objective To specifically extract and analyze nascent proteins synthesized by bone marrow mesenchymal stem cells(BMSCs)after transplantation into ischemic hearts using a technique employing mutant methionyl-tRNA synthetase(MetRSL247G)for nascent protein labeling,in order to explore the potential mechanisms of action in BMSCs post-transplantation.Methods Point mutation at position 274 of the MetRS gene in BMSCs was induced via lentiviral infection to enable azidonorleucine(ANL)-mediated labeling of nascent proteins in BMSCs.The labeling efficiency was verified by means of fluorescent non-canonical amino-acid tagging(FUNCAT).Thirty healthy female C57BL/6J mice(8-10 weeks old)were divided into control and experimental groups,with 15 mice in each group.The acute myocardial infarction model was constructed by ligating the left anterior descending coronary artery in experimental group,while control mice underwent only thoracotomy without coronary ligation.After modeling,both groups received intramyocardial injections of MetRSL247G-modified BMSCs(MetRSL247G-BMSCs)at 3 different sites in the peri-infarct ischemic region.Mice were intraperitoneally injected with ANL every 6 hours for 4 times on postoperative days 0,2,and 6(n=5 for each time point)respectively,euthanized 24 h after the last injection,and cardiac tissues were isolated.The newly synthesized and labeled proteins produced by BMSCs after transplantation into the myocardium of experimental and control groups were collected,using an enrichment technique for ANL-tagged proteins and liquid chromatography-tandem mass spectrometry(LC-MS)analysis.Gene ontology(GO)analysis,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis,protein-protein interaction(PPI)analysis,and heatmap visualization analysis were performed to identify differentially expressed proteins at the 3 time points and screen key pathways and genes.Results Under fluorescence microscopy,the MetRSL247G lentivirus-infected BMSCs were observed to be labelled with mCherry signals,confirming the successful construction of the MetRSL247G-BMSCs cell line.Green fluorescent signals were detected only in nascent proteins in culture medium containing both MetRSL247G-BMSCs and ANL,validating the sensitivity and specificity of the labeling method.GO analysis revealed that differentially expressed proteins were primarily involved in basic cellular biological processes such as extracellular exosome formation,extracellular matrix organization,and focal adhesion.KEGG and PPI analyses indicated that the differential proteins were mainly involved in complement and coagulation cascade pathway,actin cytoskeleton regulation pathway,and apoptosis pathway.Heatmap analysis showed significantly upregulated expression of anti-apoptosis and cell adhesion-related factors in experimental group on day 1(P＜0.05),upregulated anti-apoptotic factors,pro-apoptotic factors,and cell adhesion-related factors on day 3(P＜0.05),and upregulated anti-apoptotic factors,cell differentiation-related factors,and cell adhesion-related factors on day 7(P＜0.05)compared with control group.Expression of apoptosis-inducing factor 1 was significantly downregulated on days 1 and 7(P＜0.05).On day 3,most differentially expressed proteins,including anti-apoptosis factors(Protein S100-A11,Clusterin,Gelsolin),pro-apoptosis factor(Cathepsin B),cell differentiation-related factor(Transgelin-2),and cell adhesion-related factors(Cofilin-1,Periostin,Fibronectin)were significantly upregulated(P＜0.05).Conclusions The MetRSL247G mutation enables BMSCs to incorporate ANL and synthesize labeled proteins,confirming the feasibility of this nascent protein labeling technique.Nascent proteins of BMSCs in ischemic myocardium primarily contribute to extracellular exosome secretion and extracellular matrix organization.BMSCs may adapt to and respond to ischemic and hypoxic environments by influencing complement and coagulation cascades,activating inflammatory factors,regulating actin cytoskeleton structure,and modulating apoptosis,thereby maintaining the survival of BMSCs.

Objective:To analyze inpatients preferences for an unaccompanied ward and its influencing factors, for references for the implementation of unaccompanied ward management.Methods:Based on a convergent mixed research design, a convenience sampling method was used to select inpatients who visited a tertiary hospital from June to August 2024 as the survey subjects. A questionnaire survey was conducted on their willingness to choose the unaccompanied ward (provided by medical nursing staff). Meanwhile, using purposive sampling method, 10 inpatients and 11 accompanying family members were selected for semi-structured interviews about unaccompanied ward prference, and the interview topics were summarized and extracted. The results of quantitative and qualitative were compared and integrated.Results:The quantitative research results showed that among the 805 inpatients included, 125 patients (13.03%) chose medical caregivers, 382 patients (39.83%) chose their spouses, and 272 patients (28.36%) chose their children; 411 patients (24.54%) did not choose medical caregivers due to financial burden; 509 patients (63.23%) believed that the cost of an unaccompanied ward should be less than 120 yuan/day. The qualitative research results showed that the interview data formed three themes, including the driving factors of caregiving form selection intention, the emotional tendency of caregiving form selection intention, and the assessment of the benefits and drawbacks of caregiving form selection intention. The mixed research results showed that the majority of inpatients choose their spouse or children accompany them, and their willingness to choose medical caregivers is mainly influenced by service costs and family labor. However, its did not affect patients who are young or have difficulty caring for them; Influenced by traditional Chinese culture, patients tended to choose relatives to accompany them; The main reasons why patients did not choose unaccompanied wards were the economic burden and the mismatch between medical nursing staff services and their expectations.Conclusions:Inpatients tended to choose family members to accompany them. Their willingness to choose unaccompanied ward was influenced by economic burden, family labor, patient age, caregiving difficulty, and filial piety culture.

Objective By observing and measuring the relevant data of the buccal branch of the facial nerve,the parotid duct and the facial artery,the positional relationship among the three was analyzed to avoid accidental injury to the buccal branch of the facial nerve and the parotid duct when ligaturing the facial artery during the operation.Methods Forty adult head and neck specimens were dissected to observe the relationship between the buccal branch of the facial nerve and the parotid duct,the course and positional relationship of the facial artery,and the relationship between the buccal branch of the facial nerve and the peripheral vascular network.The relevant diameters were measured with a vernier caliper.Results The buccal branch of the facial nerve was divided into the superior buccal branch and the inferior buccal branch,and there was no direct anastomosis or connecting fiber between the buccal branch of the facial nerve and the parotid duct.The superior buccal branch was relatively thick,and it has a relatively constant position,which was parallel to the parotid duct.The position of the inferior buccal branch was not constant and it ran on or slightly above the plane of angulus oris.The superior buccal branch was located(10.76±5.54)mm from the parotid duct,while the inferior buccal branch was positioned(6.84±4.06)mm away from the parotid duct.The course of the main trunk of the facial artery was relatively fixed.Moreover,if the branch of the facial artery was missing,other branches of the facial artery would extend to replace the missing branch artery.The main trunk of the facial artery had a diameter of(2.34±0.83)mm,and its branches formed anastomoses with the buccal branch of the maxillary artery,creating a vascular network in the parotid and buccal regions.There was a vascular network around the buccal branch of the facial nerve,which was mostly small branches of the facial artery and the superficial temporal artery.Conclusion The buccal branch of the facial nerve exhibits a consistent anatomic relationship with the parotid duct and the facial artery.During the ligation of the facial artery,the parotid duct can serve as a landmark to accurately locate the buccal branch of the facial nerve,thereby significantly reducing the risk of inadvertent injury to the buccal branch of the facial nerve and the parotid duct.

Objective To explore the effects of different concentrations of simvastatin on nerve regeneration after sciatic nerve injury.Methods Rats were randomly divided into the normal group,the control group,the low-dose group and the high-dose group,with 3 rats in each group.Except for the normal group,adult rat sciatic nerve crush injury models were established in the other groups.Rats in the normal group and the control group were orally administered with water,while those in the low-dose group and high-dose group were orally administered with 98%simvastatin at dosages of 4 mg/mL and 40 mg/mL,respectively.The sciatic nerve regeneration in rats was evaluated by sciatic function index(SFI),HE staining,luxol fast blue(LFB)staining and immunofluorescence staining,etc.Results The SFI of rats in the high-dose group 7 days and 14 days after surgery were higher than those in the control group(P<0.05);there was no significant difference in SFI of rats between the low-dose group and the control group 7 days and 14 days after surgery(P>0.05).HE staining and LFB staining results showed that compared with the control group,the number of neurons of rats in the high-dose group increased,the nerve fibers and myelin were clearer and denser,and the nerve function was significantly restored;while no significant improvement was observed in the sciatic nerve of rats in the low-dose group.The immunofluorescence staining results showed that compared with the control group,the immunofluorescence intensity in the high-dose group increased,while that in the low-dose group decreased,the differences were statistically significant(P<0.05).Conclusion High-dose simvastatin can promote peripheral nerve regeneration by regulating the expression of M2 macrophages.

Objective·To develop a nomogram prediction model and an online risk calculator,and to predict the continence of patients after robot-assisted laparoscopic radical prostatectomy(RARP).Methods·A total of 604 prostate cancer patients who underwent RARP and had preoperative prostate magnetic resonance imaging at the Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine from September 2022 to December 2024 were analyzed and included.All patients were randomly resampled and divided into a training set(n=420)and a validation set(n=184)at a ratio of 7∶3.The patients'continence was followed up every month from the first month after the operation.The least absolute shrinkage and selection operator(LASSO)model was applied to screen the features.A Logistic multivariate regression analysis was used to establish a prediction model integrating the features selected from the LASSO analysis.The receiver operator characteristic(ROC)curve was drawn to predict the recovery of continence in patients after RARP,and the areas under the curve were compared by the DeLong test to evaluate the discrimination of the model.Calibration curves and decision curve analysis(DCA)were used to evaluate the calibration and clinical utility the model.Results·According to the postoperative continence follow-up data of the patients,the continence rate of the patients at 3 months after the operation was 58.28%(352/604).The length of the membranous urethra,the thickness of the right levator ani muscle,and blood loss were identified as independent predictors of early postoperative(3-month)incontinence by Logistic multivariate regression analysis of the training set.The area under the ROC curve was calculated as 0.976(0.954,0.998)for the training set and 0.977(0.945,1.000)for the validation set,demonstrating good discrimination of this model.No significant difference between the ROC curves of the training set and the validation set was confirmed by the DeLong test(P=0.949).A good goodness of fit of this model was demonstrated by the Hosmer-Lemeshow test(P=0.179).The clinical utility of the nomogram prediction model was indicated by the DCA plot.This nomogram prediction model was incorporated into an online calculator(https://yitingd.shinyapps.io/DynNomapp).Conclusion·This study developed and validated a nomogram prediction model that can effectively predict the early continence of patients after RARP.The length of the membranous urethra,the thickness of the right levator ani muscle,and the intraoperative blood loss are significant independent predictors of early postoperative incontinence.

Objective:To investigate the effect of extracorporeal membrane oxygenation (ECMO) in assisting neonates with complex congenital heart diseases (CHD) and the survival rate of the patients.Methods:A retrospective case-control study was made on 22 newborns with complex CHD assisted by ECMO during the perioperative period in Fuwai Central China Cardiovascular Hospital from January 2018 to June 2024.There were 19 males and 3 females in the newborns included, with an average age of (10.4±8.7) days (range: 1-26 days) and an average weight of (3.1±0.3) kg.Complex CHD included total anomalous pulmonary venous connection in 9 cases (40.9%), interrupted aortic arch and coarctation in 8 cases (36.4%), transposition of great arteries in 3 cases (13.6%), double outlet right ventricle in 1 case (4.5%) and cardiac tumor in 1 case (4.5%).The patients were divided into the <5 d group, 5-12 d group, and >12 d group according to the duration of ECMO support.Data were compared using the t-test or χ2 test. Results:There were 8 cases (36.4%) successfully weaned from ECMO support and 6 cases (27.3%) survived.ECMO support was used routinely in 15 cases (68.2%), and for extracorporeal cardiopulmonary resuscitation in 7 cases (31.8%).The duration of ECMO support was (5.9±3.7) days.There were no statistically differences in age, weight, gender, disease composition, operation time, cardiopulmonary bypass time, cross-clamp time, assistant time, blood routine, liver and kidney function and other biochemical indicators, preoperative cardiac ejection fraction value, procalcitonin, C-reactive protein and other infection indicators between the death group and the survival group (all P>0.05).The highest lactate values 24 hours before [(8.1±5.4) mmol/L] and after ECMO support [(10.5±7.1) mmol/L] in the survival group were significantly lower than those in the death group [(18.7±9.2) mmol/L, (21.3±8.6) mmol/L] ( t=2.606, P=0.018; t=2.729, P=0.013).It was found that the survival rate was 0/9 (0) in the <5 d group, 6/12 (50.0%) in the 5-12 d group, and 0/1 (0) in the >12 d group.The survival rate of the 5-12 d group was the highest, which was significantly higher than that of the <5 d group ( χ2=6.300, P=0.012). Conclusions:ECMO support is an effective treatment for severe circulatory failure in neonates with complex CHD in the perioperative period.The highest lactate levels 24 hours before and after ECMO support affect the survival rate.Patients receiving 5-12 d ECMO support can achieve the highest survival rate.

Objective·To develop a nomogram prediction model and an online risk calculator,and to predict the continence of patients after robot-assisted laparoscopic radical prostatectomy(RARP).Methods·A total of 604 prostate cancer patients who underwent RARP and had preoperative prostate magnetic resonance imaging at the Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine from September 2022 to December 2024 were analyzed and included.All patients were randomly resampled and divided into a training set(n=420)and a validation set(n=184)at a ratio of 7∶3.The patients'continence was followed up every month from the first month after the operation.The least absolute shrinkage and selection operator(LASSO)model was applied to screen the features.A Logistic multivariate regression analysis was used to establish a prediction model integrating the features selected from the LASSO analysis.The receiver operator characteristic(ROC)curve was drawn to predict the recovery of continence in patients after RARP,and the areas under the curve were compared by the DeLong test to evaluate the discrimination of the model.Calibration curves and decision curve analysis(DCA)were used to evaluate the calibration and clinical utility the model.Results·According to the postoperative continence follow-up data of the patients,the continence rate of the patients at 3 months after the operation was 58.28%(352/604).The length of the membranous urethra,the thickness of the right levator ani muscle,and blood loss were identified as independent predictors of early postoperative(3-month)incontinence by Logistic multivariate regression analysis of the training set.The area under the ROC curve was calculated as 0.976(0.954,0.998)for the training set and 0.977(0.945,1.000)for the validation set,demonstrating good discrimination of this model.No significant difference between the ROC curves of the training set and the validation set was confirmed by the DeLong test(P=0.949).A good goodness of fit of this model was demonstrated by the Hosmer-Lemeshow test(P=0.179).The clinical utility of the nomogram prediction model was indicated by the DCA plot.This nomogram prediction model was incorporated into an online calculator(https://yitingd.shinyapps.io/DynNomapp).Conclusion·This study developed and validated a nomogram prediction model that can effectively predict the early continence of patients after RARP.The length of the membranous urethra,the thickness of the right levator ani muscle,and the intraoperative blood loss are significant independent predictors of early postoperative incontinence.

Objective By observing and measuring the relevant data of the buccal branch of the facial nerve,the parotid duct and the facial artery,the positional relationship among the three was analyzed to avoid accidental injury to the buccal branch of the facial nerve and the parotid duct when ligaturing the facial artery during the operation.Methods Forty adult head and neck specimens were dissected to observe the relationship between the buccal branch of the facial nerve and the parotid duct,the course and positional relationship of the facial artery,and the relationship between the buccal branch of the facial nerve and the peripheral vascular network.The relevant diameters were measured with a vernier caliper.Results The buccal branch of the facial nerve was divided into the superior buccal branch and the inferior buccal branch,and there was no direct anastomosis or connecting fiber between the buccal branch of the facial nerve and the parotid duct.The superior buccal branch was relatively thick,and it has a relatively constant position,which was parallel to the parotid duct.The position of the inferior buccal branch was not constant and it ran on or slightly above the plane of angulus oris.The superior buccal branch was located(10.76±5.54)mm from the parotid duct,while the inferior buccal branch was positioned(6.84±4.06)mm away from the parotid duct.The course of the main trunk of the facial artery was relatively fixed.Moreover,if the branch of the facial artery was missing,other branches of the facial artery would extend to replace the missing branch artery.The main trunk of the facial artery had a diameter of(2.34±0.83)mm,and its branches formed anastomoses with the buccal branch of the maxillary artery,creating a vascular network in the parotid and buccal regions.There was a vascular network around the buccal branch of the facial nerve,which was mostly small branches of the facial artery and the superficial temporal artery.Conclusion The buccal branch of the facial nerve exhibits a consistent anatomic relationship with the parotid duct and the facial artery.During the ligation of the facial artery,the parotid duct can serve as a landmark to accurately locate the buccal branch of the facial nerve,thereby significantly reducing the risk of inadvertent injury to the buccal branch of the facial nerve and the parotid duct.

Objective To explore the effects of different concentrations of simvastatin on nerve regeneration after sciatic nerve injury.Methods Rats were randomly divided into the normal group,the control group,the low-dose group and the high-dose group,with 3 rats in each group.Except for the normal group,adult rat sciatic nerve crush injury models were established in the other groups.Rats in the normal group and the control group were orally administered with water,while those in the low-dose group and high-dose group were orally administered with 98%simvastatin at dosages of 4 mg/mL and 40 mg/mL,respectively.The sciatic nerve regeneration in rats was evaluated by sciatic function index(SFI),HE staining,luxol fast blue(LFB)staining and immunofluorescence staining,etc.Results The SFI of rats in the high-dose group 7 days and 14 days after surgery were higher than those in the control group(P<0.05);there was no significant difference in SFI of rats between the low-dose group and the control group 7 days and 14 days after surgery(P>0.05).HE staining and LFB staining results showed that compared with the control group,the number of neurons of rats in the high-dose group increased,the nerve fibers and myelin were clearer and denser,and the nerve function was significantly restored;while no significant improvement was observed in the sciatic nerve of rats in the low-dose group.The immunofluorescence staining results showed that compared with the control group,the immunofluorescence intensity in the high-dose group increased,while that in the low-dose group decreased,the differences were statistically significant(P<0.05).Conclusion High-dose simvastatin can promote peripheral nerve regeneration by regulating the expression of M2 macrophages.

Objective:To investigate the effect of extracorporeal membrane oxygenation (ECMO) in assisting neonates with complex congenital heart diseases (CHD) and the survival rate of the patients.Methods:A retrospective case-control study was made on 22 newborns with complex CHD assisted by ECMO during the perioperative period in Fuwai Central China Cardiovascular Hospital from January 2018 to June 2024.There were 19 males and 3 females in the newborns included, with an average age of (10.4±8.7) days (range: 1-26 days) and an average weight of (3.1±0.3) kg.Complex CHD included total anomalous pulmonary venous connection in 9 cases (40.9%), interrupted aortic arch and coarctation in 8 cases (36.4%), transposition of great arteries in 3 cases (13.6%), double outlet right ventricle in 1 case (4.5%) and cardiac tumor in 1 case (4.5%).The patients were divided into the <5 d group, 5-12 d group, and >12 d group according to the duration of ECMO support.Data were compared using the t-test or χ2 test. Results:There were 8 cases (36.4%) successfully weaned from ECMO support and 6 cases (27.3%) survived.ECMO support was used routinely in 15 cases (68.2%), and for extracorporeal cardiopulmonary resuscitation in 7 cases (31.8%).The duration of ECMO support was (5.9±3.7) days.There were no statistically differences in age, weight, gender, disease composition, operation time, cardiopulmonary bypass time, cross-clamp time, assistant time, blood routine, liver and kidney function and other biochemical indicators, preoperative cardiac ejection fraction value, procalcitonin, C-reactive protein and other infection indicators between the death group and the survival group (all P>0.05).The highest lactate values 24 hours before [(8.1±5.4) mmol/L] and after ECMO support [(10.5±7.1) mmol/L] in the survival group were significantly lower than those in the death group [(18.7±9.2) mmol/L, (21.3±8.6) mmol/L] ( t=2.606, P=0.018; t=2.729, P=0.013).It was found that the survival rate was 0/9 (0) in the <5 d group, 6/12 (50.0%) in the 5-12 d group, and 0/1 (0) in the >12 d group.The survival rate of the 5-12 d group was the highest, which was significantly higher than that of the <5 d group ( χ2=6.300, P=0.012). Conclusions:ECMO support is an effective treatment for severe circulatory failure in neonates with complex CHD in the perioperative period.The highest lactate levels 24 hours before and after ECMO support affect the survival rate.Patients receiving 5-12 d ECMO support can achieve the highest survival rate.