BACKGROUND:Previous studies have demonstrated that the cannabinoid type Ⅰ receptor can enhance the proliferation and neural differentiation of neural stem cells and mesenchymal stem cells.Moreover,cannabinoid type Ⅰ also governs the proliferation and mineralization capacity of human apical papilla stem cells.However,there are relatively few investigations concerning the impact of cannabinoid type Ⅰ overexpression on the neural differentiation of human apical papilla stem cells.OBJECTIVE:To investigate the effect of cannabinoid type Ⅰ on neural differentiation of human apical papilla stem cells in vitro.METHODS:Healthy third molars with immature root tips that need to be removed for orthodontic treatment were collected,and human apical papilla stem cells were isolated and cultured by tissue block method combined with enzyme digestion method.Cannabinoid type Ⅰ gene was introduced into human apical papilla stem cells by lentivirus-mediated transfection technique.A blank control group,a negative control group,and cannabinoid type Ⅰ overexpression group were set up.The transfection effect of overexpression of cannabinoid type Ⅰ lentivirus on human apical papilla stem cells was verified by Western Blot.The control group,negative control group,cannabinoid type Ⅰ overexpression group and cannabinoid type Ⅰ overexpression+AM251(cannabinoid type Ⅰ receptor antagonist)group were set up.Cell proliferation was detected by CCK-8 assay at 1,5,and 10 days after neural induction.On day 10 of neural induction,the expression levels of TH,NeuroD-1,and NCAM1 genes were detected by qRT-PCR,and the protein expression levels of Nestin and TUBB3 were detected by immunofluorescence.RESULTS AND CONCLUSION:(1)Compared with the blank control group and the negative control group,the expression of cannabinoid receptor Ⅰ protein in the cannabinoid receptor Ⅰ overexpression group was significantly increased,and the difference was significant(P＜0.05).(2)Compared with the blank control group and the negative control group,the proliferation ability of human apical papilla stem cells in the cannabinoid type Ⅰ overexpression group was the strongest at 5 and 10 days after neural induction(P＜0.05).(3)Compared with the blank control group and the negative control group,the mRNA expression of NeuroD-1,NCAM1,and TH in the stem cells of the human apical papilla in the cannabinoid type Ⅰ overexpression group was significantly increased,and the fluorescence intensity of Nestin and TUBB3 was significantly enhanced(P＜0.05).(4)Compared with the cannabinoid type Ⅰ overexpression group,the proliferation ability,mRNA expression level of NeuroD-1,NCAM1,and TH,as well as the fluorescence intensity of Nestin and TUBB3,were significantly decreased in the cannabinoid type Ⅰ overexpression+AM251 group(P＜0.05).These findings conclude that overexpression of cannabinoid type Ⅰ promoted the proliferation and neural differentiation of human apical dentin papilla stem cells.

BACKGROUND:Previous studies have demonstrated that the cannabinoid type Ⅰ receptor can enhance the proliferation and neural differentiation of neural stem cells and mesenchymal stem cells.Moreover,cannabinoid type Ⅰ also governs the proliferation and mineralization capacity of human apical papilla stem cells.However,there are relatively few investigations concerning the impact of cannabinoid type Ⅰ overexpression on the neural differentiation of human apical papilla stem cells.OBJECTIVE:To investigate the effect of cannabinoid type Ⅰ on neural differentiation of human apical papilla stem cells in vitro.METHODS:Healthy third molars with immature root tips that need to be removed for orthodontic treatment were collected,and human apical papilla stem cells were isolated and cultured by tissue block method combined with enzyme digestion method.Cannabinoid type Ⅰ gene was introduced into human apical papilla stem cells by lentivirus-mediated transfection technique.A blank control group,a negative control group,and cannabinoid type Ⅰ overexpression group were set up.The transfection effect of overexpression of cannabinoid type Ⅰ lentivirus on human apical papilla stem cells was verified by Western Blot.The control group,negative control group,cannabinoid type Ⅰ overexpression group and cannabinoid type Ⅰ overexpression+AM251(cannabinoid type Ⅰ receptor antagonist)group were set up.Cell proliferation was detected by CCK-8 assay at 1,5,and 10 days after neural induction.On day 10 of neural induction,the expression levels of TH,NeuroD-1,and NCAM1 genes were detected by qRT-PCR,and the protein expression levels of Nestin and TUBB3 were detected by immunofluorescence.RESULTS AND CONCLUSION:(1)Compared with the blank control group and the negative control group,the expression of cannabinoid receptor Ⅰ protein in the cannabinoid receptor Ⅰ overexpression group was significantly increased,and the difference was significant(P＜0.05).(2)Compared with the blank control group and the negative control group,the proliferation ability of human apical papilla stem cells in the cannabinoid type Ⅰ overexpression group was the strongest at 5 and 10 days after neural induction(P＜0.05).(3)Compared with the blank control group and the negative control group,the mRNA expression of NeuroD-1,NCAM1,and TH in the stem cells of the human apical papilla in the cannabinoid type Ⅰ overexpression group was significantly increased,and the fluorescence intensity of Nestin and TUBB3 was significantly enhanced(P＜0.05).(4)Compared with the cannabinoid type Ⅰ overexpression group,the proliferation ability,mRNA expression level of NeuroD-1,NCAM1,and TH,as well as the fluorescence intensity of Nestin and TUBB3,were significantly decreased in the cannabinoid type Ⅰ overexpression+AM251 group(P＜0.05).These findings conclude that overexpression of cannabinoid type Ⅰ promoted the proliferation and neural differentiation of human apical dentin papilla stem cells.

The investigator-initiated trials （IIT）， as a widely existing form of clinical research both domestically and internationally， have attracted the attention of governments around the world due to the potential legal risks and issues they may cause， and the trend of legal supervision has gradually strengthened. The legal regulation of IIT in China is still in its early stages， with numerous legal issues that need to be clarified and sorted out. Based on the domestic and international legal reviews of IIT， this paper systematically sorted out the current situation of legal supervision of IIT in China and examined the existing issues， including weak legislative rules， incomplete regulatory systems， imperfect protection rules for research participants and informed consent systems， inadequate regulation of conflicts of interest management， and lax legal supervision of ethical review. Furthermore， this paper proposed suggestions for legal regulation of IIT from three perspectives， including strengthening legislation and emphasizing regulation， improving the mechanism for protecting research participants’ rights and interests， and balancing the legalization of IIT and the scientific development of clinical research， with a view to providing references for legal regulatory paths of IIT in China.

Objective:To investigate the control status of out-of-hospital blood glucose and blood lipids in patients with acute myocardial infarction (AMI) complicated with diabetes mellitus and its correlation with prognosis.Methods:The clinical data of 406 patients with AMI complicated with diabetes mellitus from January 2017 to December 2022 in Xuanwu Hospital, Capital Medical University were retrospectively analyzed. The demographic and out-of-hospital clinical information of patients were recorded, and the control level of out-of-hospital risk factors and the occurrence of major adverse cardiovascular event (MACCE) were also recorded. The patients were grouped according to the levels of glycosylated hemoglobin (HbA 1c) and low-density lipoprotein cholesterol (LDL-C). HbA 1c<6.0% was the low HbA 1c group, HbA 1c 6.0% to 7.0% was the medium HbA 1c group, and HbA 1c>7.0% was the high HbA 1c group; LDL-C<1.4 mmol/L was low LDL-C group, LDL-C 1.4 to 1.8 mmol/L was medium LDL-C group, and LDL-C>1.8 mmol/L was high LDL-C group. Multivariate Cox regression analysis was used to analyze the independent risk factors for out-of-hospital MACCE in patients with AMI complicated with diabetes mellitus. Results:The HbA 1c data of 249 patients were recorded in detail, and only 51.0% (127/249) of patients with HbA 1c≤7%. There were statistical differences in the history of cerebral infarction, out-of-hospital fasting blood glucose, out-of-hospital total cholesterol (TC) and out-of-hospital LDL-C among the low HbA 1c group (24 cases), medium HbA 1c group (103 cases) and high HbA 1c group (122 cases) ( P<0.05). The incidences of out-of-hospital MACCE in low HbA 1c group, medium HbA 1c group and high HbA 1c group were 20.8%(5/24), 12.6%(13/103) and 32.0%(39/122), respectively. The incidence of out-of-hospital MACCE in high HbA 1c group was significantly higher than that in medium HbA 1c group, and there was statistical difference ( P<0.05); there was no statistical difference between low HbA 1c group and high HbA 1c group ( P>0.05). Among the 406 patients, 53.4%(217/406) had LDL-C≤1.8 mmol/L, and only 20.0%(81/406) had LDL-C<1.4 mmol/L. There were statistical differences in hyperlipidemia, out-of-hospital HbA 1c, out-of-hospital fasting blood glucose, out-of-hospital alanine aminotransferase (ALT), out-of-hospital TC and out-of-hospital triglyceride (TG) among low LDL-C group (81 cases), medium LDL-C group (136 cases) and high LDL-C group (189 cases) ( P<0.05). The incidences of MACCE in low LDL-C group, medium LDL-C group and high LDL-C group were 18.5% (15/81), 25.7% (35/136) and 36.5% (69/189), respectively. The incidence of MACCE in high LDL-C group was significantly higher than that in low LDL-C group, and there was statistical difference ( P<0.05); there was no statistical difference between low LDL-C group and medium LDL-C group ( P>0.05). In the different HbA 1c groups, multivariate Cox regression analysis result showed that HbA 1c>7% and high out-of-hospital fasting blood glucose were independent risk factors for out-of-hospital MACCE in patients with AMI complicated with diabetes mellitus ( OR = 2.575 and 1.064, 95% CI 1.345 to 4.927 and 1.005 to 1.128, P<0.01 and <0.05). In different LDL-C groups, multivariate Cox regression analysis result showed that high out-of-hospital HbA 1c was an independent risk factor for out-of-hospital MACCE in patients with AMI complicated with diabetes mellitus ( OR = 1.303, 95% CI 1.144 to 1.485, P<0.01). Conclusions:The control rates of out-of-hospital blood glucose and blood lipids are low in patients with AMI complicated with diabetes mellitus, and HbA 1c level can independently predict the risk of out-of-hospital MACCE in patients with AMI complicated with diabetes mellitus.

Objective:To compare the efficacy of low-dose febuxostat and low-dose benzbromarone in lowering serum uric acid and their impact on liver function in male patients with renal underexcretion gout.Methods:This prospective cohort study enrolled 303 patients with renal underexcretion gout and normal baseline liver function. Participants were assigned to either the low-dose febuxostat group(20 mg qd) or the low-dose benzbromarone group(25 mg qd). A linear mixed-effects model was used to compare the uric acid target achievement rate(<360 μmol/L) and changes in liver enzyme levels between the two groups.Results:At week 4, the proportion of patients achieving the serum uric acid target(<360 μmol/L) was significantly higher in the benzbromarone group than that in the febuxostat group(64.2% vs 42.3%, P<0.001), with a trend toward greater efficacy at weeks 8 and 12. The incidence of alanine aminotransferase(ALT) or aspartate aminotransferase(AST) elevation above the upper limit was significantly higher in the febuxostat group compared to the benzbromarone group(35.2% vs 13.85%, P<0.001). After adjusting for baseline liver enzyme levels in the mixed-effects model, mean ALT and AST levels remained significantly higher in the febuxostat group than those in the benzbromarone group at weeks 4, 8, and 12( P<0.05). In the febuxostat group, ALT and AST levels significantly increased over time during weeks 0-4 and 4-8 ( P<0.001), peaking at week 8 followed by a decreasing trend. By week 12, the levels were not significantly different from baseline ( P>0.05). Whereas there was no significant difference at each follow-up time point in the benzbromarone group( P>0.05). Conclusions:In male patients with renal underexcretion gout, low-dose benzbromarone demonstrated superior urate-lowering efficacy and better hepatic safety compared to low-dose febuxostat.

Objective:To evaluate the effect of intraoperative optimization of regional cerebral oxygen saturation(rSO 2C) intervention on postoperative delirium(POD) in pediatric patients undergoing cardiac surgery with cardiopulmonary bypass(CPB). Methods:Two hundred and seventy-three pediatric patients of both sexes, aged 28 days-6 yr, with American Society of Anesthesiologists Physical Status classification ≤Ⅳ, scheduled for elective cardiac surgery under CPB, were divided into intervention group( n=136) and control group( n=137) based on the computer random coding. In intervention group, optimized intervention measures were given when rSO 2C was below 75% of the baseline value for more than 1 min. In control group, rSO 2C was not monitored during operation, and intraoperative management was performed according to the routine monitoring indicators of pediatric cardiac surgery under CPB. The occurrence of POD within 7 days after operation was evaluated, and the duration and first occurrence time of POD were recorded. Results:Compared with control group, no significant change was found in the incidence of POD( P>0.05), the first occurrence time of POD was significantly prolonged, and the duration of POD was shortened in intervention group( P<0.05). Conclusions:Intraoperative optimization of rSO 2C intervention can delay the time to the first occurrence of POD and shorten the duration in pediatric patients undergoing cardiac surgery under CPB.

Objective:To evaluate the association between postoperative delirium (POD) and preoperative frailty in elderly patients undergoing knee or hip arthroplasty.Methods:This nested case-control study utilized medical records from elderly patients who underwent knee or hip arthroplasty under combined spinal-epidural anesthesia at Qingdao Municipal Hospital between September 2021 and May 2023. Participants were divided into 2 groups based on clinically diagnosed POD: POD group ( n=53) and non-POD group ( n=256). Univariate analysis was conducted on suspected influencing factors, and logistic regression analysis was utilized to identify the risk factors for POD. Receiver operating characteristic and clinical decision curves were plotted to evaluate the predictive performance of these risk factors for POD. Mediation analysis was performed, and a clinically applicable nomogram was constructed to achieve visual prediction of outcomes. Results:There were statistically significant differences in age, preoperative frailty, body mass index, American Society of Anesthesiologists Physical Status classification, Memorial Delirium Assessment Scale scores, and concentrations of Aβ 42, Aβ 40, phosphorylated tau protein (p-tau protein) and tau protein, Aβ 42/tau ratio and Aβ 42/p-tau ratio in cerebrospinal fluid (CSF) between non-POD group and POD group ( P<0.05). Preoperative frailty was a risk factor for POD ( P<0.05). Mediation analysis revealed that the association between preoperative frailty and POD was mediated by CSF tau protein concentrations. The area under the receiver operating characteristic curve of preoperative frailty and CSF biomarker concentrations in predicting POD was 0.974 ( P<0.05). The clinical decision curve demonstrated that the model combining the preoperative frailty and CSF biomarker concentrations predicted a higher net benefit ( P<0.05). The clinical decision curve showed that the model combining preoperative frailty and CSF biomarker concentrations predicted a higher net benefit. Conclusions:Preoperative frailty is a risk factor for POD in elderly patients undergoing knee or hip arthroplasty, and its combination with CSF biomarker concentrations can effectively predict the occurrence of POD. CSF tau concentration mediates the association between preoperative frailty and development of POD.

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 observe the efficacy and safety of mechanical thrombectomy in acute ischemic stroke patients after vertebral-basilar artery stent implantation.Methods From May 2018 to October 2022,patients with acute vertebral basilar artery occlusion who underwent mechanical thrombectomy were retrospectively analyzed.The patients were divided into two groups:stent occlusion group and acute atherosclerotic stenosis occlusion(ICAS)group.The baseline characteristics of the two groups were balanced by propensity score matching method.The successful recanalization rate,90-day good outcome rate,90-day mortality rate,and symptomatic intracranial hemorrhage rate were compared.Results We enrolled 107 patients,with 14 cases in stent occlusion group and 93 cases in ICAS group.We analyzed 14 pairs by propensity score matching,including 14 cases in stent occlusion group and 27 cases in ICAS group.The successful recanalization rate and 90-day good outcome rate was lower in stent occlusion group than in ICAS group[(78.6％(11/14)vs.100％(27/27),P=0.062,28.6％(4/14)vs.44.4％(12/27),X2=0.976,P=0.323],but there was no statistical difference.The 90-day mortality rate in the stent occlusion group was significantly higher than that in the ICAS group[57.1％(8/14)vs.25.9％(7/27),x2=3.873,P=0.049].The incidence of symptomatic intracranial hemorrhage was higher in the stent occlusion group than in the ICAS group,with no statistical difference[35.7％(5/14)vs.14.8％(4/27),x2=1.289,P=0.256].Conclusion The successful recanalization rate,90-day good outcome rate,and incidence of symptomatic intracranial hemorrhage in stent occlusion group did not significantly differ from those in ICAS group,but the 90-day mortality rate was significantly higher in the former group.

Objective To explore the efficiency of operating room(OR)and its influencing factors from a phenomeno-logical perspective,proposing corresponding improvement strategies.Methods A phenomenological method was adopted,invol-ving in-depth interviews with 13 surgical team members from a tertiary Traditional Chinese Medicine(TCM)orthopedic hospital in Henan Province,to collect and analyze their multifaceted understanding of OR efficiency.Results The study demonstrated eight interpretative ways of efficiency among team members.The overall framework was largely organization-oriented,with addi-tional human-centered considerations regarding the factors influencing efficiency.Conclusion A unified understanding is crucial for improving both efficiency and quality.This study suggests strategies to optimize OR processes through lean management and human-centered team development to enhance operating room efficiency.Additionally,it highlights the need for future research to incorporate more patient views.