Objective:To investigate the role of microRNA-25-5p (miR-25-5p) in melanogenesis, and to explore its underlying mechanisms.Methods:Target genes of miR-25-5p were predicted using the TargetScan database. The interaction between miR-25-5p and the 3' untranslated region (3' UTR) of the RAB11B gene (a member of RAS oncogene family) was validated through a dual-luciferase reporter assay. Post-inflammatory hyperpigmentation (PIH) models were established in female C57BL/6J mice (6 - 8 weeks old) and female brown guinea pigs (4 - 6 weeks old) through daily broadband ultraviolet B (UVB) irradiation on the dorsal skin of the mouse ear or shaved dorsal skin of guinea pigs, while untreated mice and untreated dorsal skin areas of guinea pigs served as control groups. During modeling, these experimental animals received intradermal injections of a miR-25-5p agomir or a miR control agomir. Changes in skin pigmentation were observed, and skin tissue samples were harvested for further analysis after modeling. Melanin content in skin tissues was evaluated using Masson-Fontana staining. Expression of RAB11B and tyrosinase (TYR) in skin tissues was determined using immunohistochemical staining and quantitative real-time PCR (qPCR). Primary human melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision. Both primary human melanocytes and human MNT1 melanoma cells were transfected with miR-25-5p mimics or miR control mimics. Relative expression levels of miR-25-5p and RAB11B mRNA were quantified by qPCR using the 2 -ΔΔCt calculation method. In MNT1 cells, miR-25-5p and RAB11B were co-overexpressed to assess their effect on the mRNA expression of RAB11B and TYR. Statistical analysis was conducted using t test or one-way analysis of variance followed by Tukey's post hoc test for multiple comparisons. Results:The bioinformatic prediction and dual-luciferase reporter assay confirmed a binding site for miR-25-5p in the 3′ UTR of the RAB11B gene. In both animal models, the treatment with the miR-25-5p agomir significantly reduced local skin pigmentation compared to the control groups; Masson-Fontana staining showed a marked decrease in the density of melanin granules in the epidermis and dermis in the miR-25-5p agomir groups compared with the miR control agomir groups (mice: 0.050 ± 0.005 vs. 0.087 ± 0.008; guinea pigs: 0.067 ± 0.015 vs. 0.110 ± 0.013; both P < 0.05). Immunohistochemical staining revealed significantly lower expression of RAB11B in mouse skin tissues in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). qPCR revealed significantly lower mRNA expression of RAB11B and TYR in skin tissues of guinea pigs in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). Similarly, RAB11B mRNA expression significantly decreased in the miR-25-5p mimics group compared with the miR control mimics group in primary human melanocytes and MNT1 cells (both P < 0.05). In human MNT1 melanoma cells, miR-25-5p overexpression could suppress TYR mRNA expression, whereas co-overexpression of miR-25-5p and RAB11B could reverse this suppression. Conclusion:Overexpression of miR-25-5p could alleviate UVB-induced post-inflammatory hyperpigmentation and inhibit melanogenesis, likely by targeted suppression of RAB11B expression.

Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Objective To construct a predictive model for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke and evaluate its predictive efficacy.Methods Patients with acute stroke caused by anterior circulation large vessel occlusion who underwent endovascular treatment with successful vascular recanalization(modified thrombolysis in cerebral infarction[mTICI]grade 2b or 3)admitted to the Department of Neurosurgery of Tianjin Union Medical Center(the First Affiliated Hospital of Nankai University)from January 2022 to January 2025 were retrospectively and consecutively included.Patients were divided into effective recanalization group(modified Rankin scale[mRS]score≤2)and futile recanalization group(mRS score＞2)according to the mRS score at 90 days after endovascular treatment.Baseline and clinical data were collected,including sex,age,medical history(stroke history,hypertension,diabetes mellitus,atrial fibrillation),personal history(smoking history,alcohol consumption history),and National Institutes of Health stroke scale(NIHSS)score,blood pressure,blood glucose,total cholesterol,triglycerides,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,creatinine,neutrophil-to-lymphocyte ratio,fibrinogen,D-dimer,lymphocyte-to-monocyte ratio,white blood cell count,hemoglobin,albumin,C-reactive protein-to-albumin ratio,platelet-to-lymphocyte ratio at admission.Imaging data including culprit vessel(middle cerebral artery,anterior cerebral artery,internal carotid artery),Alberta stroke program early CT score(ASPECTS),infarct core volume,hypoperfusion area volume,mismatch volume between infarct core and hypoperfusion area,American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology(ASITN/SIR)collateral circulation grading,trial of Org 10172 in acute stroke treatment(TOAST)classification.Surgery-related data including onset-to-puncture time,successful puncture-to-vascular recanalization time,immediate postoperative mTICI grade,complications within 24 hours postoperatively(fever,hemorrhagic transformation,symptomatic intracranial hemorrhage),immediate postoperative mTICI grade.Factors with P＜0.05 and without multicollinearity(variance inflation factor≥5)in univariate analysis were included in multivariate Logistic regression model analysis to analyze independent influencing factors for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke.A nomogram prediction model was constructed based on statistically significant factors in multivariate Logistic regression analysis,and Hosmer-Lemeshow test was used to assess the goodness of fit of the model.The receiver operating characteristic(ROC)curve of the nomogram prediction model for predicting futile recanalization after endovascular treatment in patients with anterior circulation large vessel occlusion stroke was drawn,and calibration curve was used to evaluate the calibration of the nomogram prediction model.Results A total of 187 patients who met the criteria of this study were included,comprising 101 males and 86 females,aged 42-85 years,with a mean age of(70±6)years.The mRS scores at 90days after treatment were:0points in 21 cases(11.23%),1 point in 37 cases(19.79%),2 points in 28 cases(14.97%),3 points in 46 cases(24.60%),4 points in 31 cases(16.58%),5 points in 24 cases(12.83%),and 6 points in 0case.There were 86cases in the effective recanalization group and 101 cases in the futile recanalization group.(1)Statistically significant differences between the two groups were observed in age,atrial fibrillation,infarct core volume,admission NIHSS score,neutrophil-to-lymphocyte ratio,lymphocyte-to-monocyte ratio,and C-reactive protein-to-albumin ratio(all P＜0.05).(2)Multicollinearity analysis showed that the variance inflation factors of all independent variables with statistically significant differences in univariate analysis were all＜5,indicating no multicollinearity.Multivariate Logistic regression analysis showed that age(OR,1.101,95%CI 1.046-1.158,P=0.037),infarct core volume(OR,1.350,95%CI 1.202-1.517,P＜0.01),admission NIHSS score(OR,1.501,95%CI 1.213-1.858,P＜0.01),lymphocyte-to-monocyte ratio(OR,0.039,95%CI 0.009-0.179,P＜0.01),and C-reactive protein-to-albumin ratio(OR,6.015,95%CI 1.625-22.257,P=0.007)were independent influencing factors for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke.(3)The receiver operating characteristic curve of the nomogram prediction model for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke was constructed based on statistically significant factors in multivariate analysis showed a specificity of 0.970,sensitivity of 0.895,precision of 0.916,recall of 0.970,and area under the curve of 0.979(95%CI0.962-0.996,P＜0.01).The absolute mean error of the calibration curve was 0.008,with predicted values close to actual values.Hosmer-Lemeshow test results showed that the model had good consistency without overfitting(χ2=4.830,P=0.776).Conclusions Age,infarct core volume,admission NIHSS score,lymphocyte-to-monocyte ratio,and C-reactive protein-to-albumin ratio are all independent influencing factors for futile recanalization after endovascular treatment in acute anterior circulation large vessel occlusion stroke.The nomogram prediction model constructed accordingly can predict the risk of futile recanalization after endovascular treatment in patients with anterior circulation large vessel occlusion stroke.

Objective:To investigate the role of microRNA-25-5p (miR-25-5p) in melanogenesis, and to explore its underlying mechanisms.Methods:Target genes of miR-25-5p were predicted using the TargetScan database. The interaction between miR-25-5p and the 3' untranslated region (3' UTR) of the RAB11B gene (a member of RAS oncogene family) was validated through a dual-luciferase reporter assay. Post-inflammatory hyperpigmentation (PIH) models were established in female C57BL/6J mice (6 - 8 weeks old) and female brown guinea pigs (4 - 6 weeks old) through daily broadband ultraviolet B (UVB) irradiation on the dorsal skin of the mouse ear or shaved dorsal skin of guinea pigs, while untreated mice and untreated dorsal skin areas of guinea pigs served as control groups. During modeling, these experimental animals received intradermal injections of a miR-25-5p agomir or a miR control agomir. Changes in skin pigmentation were observed, and skin tissue samples were harvested for further analysis after modeling. Melanin content in skin tissues was evaluated using Masson-Fontana staining. Expression of RAB11B and tyrosinase (TYR) in skin tissues was determined using immunohistochemical staining and quantitative real-time PCR (qPCR). Primary human melanocytes were isolated from discarded normal foreskin tissues of healthy males after circumcision. Both primary human melanocytes and human MNT1 melanoma cells were transfected with miR-25-5p mimics or miR control mimics. Relative expression levels of miR-25-5p and RAB11B mRNA were quantified by qPCR using the 2 -ΔΔCt calculation method. In MNT1 cells, miR-25-5p and RAB11B were co-overexpressed to assess their effect on the mRNA expression of RAB11B and TYR. Statistical analysis was conducted using t test or one-way analysis of variance followed by Tukey's post hoc test for multiple comparisons. Results:The bioinformatic prediction and dual-luciferase reporter assay confirmed a binding site for miR-25-5p in the 3′ UTR of the RAB11B gene. In both animal models, the treatment with the miR-25-5p agomir significantly reduced local skin pigmentation compared to the control groups; Masson-Fontana staining showed a marked decrease in the density of melanin granules in the epidermis and dermis in the miR-25-5p agomir groups compared with the miR control agomir groups (mice: 0.050 ± 0.005 vs. 0.087 ± 0.008; guinea pigs: 0.067 ± 0.015 vs. 0.110 ± 0.013; both P < 0.05). Immunohistochemical staining revealed significantly lower expression of RAB11B in mouse skin tissues in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). qPCR revealed significantly lower mRNA expression of RAB11B and TYR in skin tissues of guinea pigs in the miR-25-5p agomir group than in those in the miR control agomir group (both P < 0.05). Similarly, RAB11B mRNA expression significantly decreased in the miR-25-5p mimics group compared with the miR control mimics group in primary human melanocytes and MNT1 cells (both P < 0.05). In human MNT1 melanoma cells, miR-25-5p overexpression could suppress TYR mRNA expression, whereas co-overexpression of miR-25-5p and RAB11B could reverse this suppression. Conclusion:Overexpression of miR-25-5p could alleviate UVB-induced post-inflammatory hyperpigmentation and inhibit melanogenesis, likely by targeted suppression of RAB11B expression.

Objective To construct a predictive model for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke and evaluate its predictive efficacy.Methods Patients with acute stroke caused by anterior circulation large vessel occlusion who underwent endovascular treatment with successful vascular recanalization(modified thrombolysis in cerebral infarction[mTICI]grade 2b or 3)admitted to the Department of Neurosurgery of Tianjin Union Medical Center(the First Affiliated Hospital of Nankai University)from January 2022 to January 2025 were retrospectively and consecutively included.Patients were divided into effective recanalization group(modified Rankin scale[mRS]score≤2)and futile recanalization group(mRS score＞2)according to the mRS score at 90 days after endovascular treatment.Baseline and clinical data were collected,including sex,age,medical history(stroke history,hypertension,diabetes mellitus,atrial fibrillation),personal history(smoking history,alcohol consumption history),and National Institutes of Health stroke scale(NIHSS)score,blood pressure,blood glucose,total cholesterol,triglycerides,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,creatinine,neutrophil-to-lymphocyte ratio,fibrinogen,D-dimer,lymphocyte-to-monocyte ratio,white blood cell count,hemoglobin,albumin,C-reactive protein-to-albumin ratio,platelet-to-lymphocyte ratio at admission.Imaging data including culprit vessel(middle cerebral artery,anterior cerebral artery,internal carotid artery),Alberta stroke program early CT score(ASPECTS),infarct core volume,hypoperfusion area volume,mismatch volume between infarct core and hypoperfusion area,American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology(ASITN/SIR)collateral circulation grading,trial of Org 10172 in acute stroke treatment(TOAST)classification.Surgery-related data including onset-to-puncture time,successful puncture-to-vascular recanalization time,immediate postoperative mTICI grade,complications within 24 hours postoperatively(fever,hemorrhagic transformation,symptomatic intracranial hemorrhage),immediate postoperative mTICI grade.Factors with P＜0.05 and without multicollinearity(variance inflation factor≥5)in univariate analysis were included in multivariate Logistic regression model analysis to analyze independent influencing factors for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke.A nomogram prediction model was constructed based on statistically significant factors in multivariate Logistic regression analysis,and Hosmer-Lemeshow test was used to assess the goodness of fit of the model.The receiver operating characteristic(ROC)curve of the nomogram prediction model for predicting futile recanalization after endovascular treatment in patients with anterior circulation large vessel occlusion stroke was drawn,and calibration curve was used to evaluate the calibration of the nomogram prediction model.Results A total of 187 patients who met the criteria of this study were included,comprising 101 males and 86 females,aged 42-85 years,with a mean age of(70±6)years.The mRS scores at 90days after treatment were:0points in 21 cases(11.23%),1 point in 37 cases(19.79%),2 points in 28 cases(14.97%),3 points in 46 cases(24.60%),4 points in 31 cases(16.58%),5 points in 24 cases(12.83%),and 6 points in 0case.There were 86cases in the effective recanalization group and 101 cases in the futile recanalization group.(1)Statistically significant differences between the two groups were observed in age,atrial fibrillation,infarct core volume,admission NIHSS score,neutrophil-to-lymphocyte ratio,lymphocyte-to-monocyte ratio,and C-reactive protein-to-albumin ratio(all P＜0.05).(2)Multicollinearity analysis showed that the variance inflation factors of all independent variables with statistically significant differences in univariate analysis were all＜5,indicating no multicollinearity.Multivariate Logistic regression analysis showed that age(OR,1.101,95%CI 1.046-1.158,P=0.037),infarct core volume(OR,1.350,95%CI 1.202-1.517,P＜0.01),admission NIHSS score(OR,1.501,95%CI 1.213-1.858,P＜0.01),lymphocyte-to-monocyte ratio(OR,0.039,95%CI 0.009-0.179,P＜0.01),and C-reactive protein-to-albumin ratio(OR,6.015,95%CI 1.625-22.257,P=0.007)were independent influencing factors for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke.(3)The receiver operating characteristic curve of the nomogram prediction model for futile recanalization after endovascular treatment in patients with acute anterior circulation large vessel occlusion stroke was constructed based on statistically significant factors in multivariate analysis showed a specificity of 0.970,sensitivity of 0.895,precision of 0.916,recall of 0.970,and area under the curve of 0.979(95%CI0.962-0.996,P＜0.01).The absolute mean error of the calibration curve was 0.008,with predicted values close to actual values.Hosmer-Lemeshow test results showed that the model had good consistency without overfitting(χ2=4.830,P=0.776).Conclusions Age,infarct core volume,admission NIHSS score,lymphocyte-to-monocyte ratio,and C-reactive protein-to-albumin ratio are all independent influencing factors for futile recanalization after endovascular treatment in acute anterior circulation large vessel occlusion stroke.The nomogram prediction model constructed accordingly can predict the risk of futile recanalization after endovascular treatment in patients with anterior circulation large vessel occlusion stroke.

Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females，with a body mass index（BMI）of（25.50 ± 2.03）kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery，and polymerase chain reaction-restriction fragment length polymorphisms（PCR-RFLP）were used to detect CYP11B2（rs1799998）promoter region 344（C > T）base substitution. The follow-up duration was more than 6 months，with the following parameters recorded at the last follow-up：plasma aldosterone，renin，serum potassium，and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score（PASO）for primary aldosteronism，and the specific criteria were as follows. ① Clinical complete remission：the patient's blood pressure returned to normal（< 140/90 mmHg，1 mmHg = 0.133 kPa）and all antihypertensive drugs were discontinued；②Partial clinical remission：blood pressure returns to normal，and the number or dose of antihypertensive drugs is reduced compared with before；③Clinical non-remission：blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis，followed by binary logistic forward conditional regression for multivariate analysis，and a variety of machine learning algorithms such as random forest，logistic regression，support vector machine and gradient lifter were integrated，and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model，and the predictive performance of the model was evaluated by using receiver operating characteristic（ROC）curve，calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2（344C > T）（rs1799998）as follows：CC type in 50 cases（43.1%），CT type in 46 cases（39.7%），and TT type in 20 cases（17.3%）. There were 74 cases in the complete remission group and 42 cases in the incomplete remission group，and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index［（24.27 ± 2.90）kg/m 2 vs.（26.98 ± 3.17）kg/m 2， P<0.001］，preoperative hypertension grade（grade 1/2/3：29/29/16 cases vs. 9/13/20 cases， P = 0.012），preoperative antihypertensive drugs（0/1/≥ 2：25/32/17 cases vs. 7/15/20 cases， P = 0.016），and CYP11B2（344C > T）（CC/TT + CT：39/35 cases vs. 11/31 cases， P = 0.006）were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs［≥ 2： OR = 5.26（95% CI 1.12?24.61， P = 0.016；1： OR = 4.55（95% CI 1.23?22.47）， P = 0.025］was the strongest independent predictor，followed by CYP11B2（344C > T）［ OR = 4.02（95% CI 1.16?13.82）， P = 0.028］and BMI［ OR = 3.96（95% CI 2.26?6.92）， P < 0.001］. Comparing the receiver operating feature（ROC）curves of the four types of machine learning models，the best model was the support vector machine model with an area under the curve（AUC）of 0.88（95% CI 0.82?0.95），followed by the gradient elevator model of 0.83（95% CI 0.76?0.91），the logistic regression model of 0.78（95% CI 0.68?0.88），and the random forest model of 0.77（95% CI 0.68?0.86）. The optimal threshold of the Yoden index of the support vector machine model was 0.588，with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms，BMI，and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients，providing new evidence for personalized treatment strategies

Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females，with a body mass index（BMI）of（25.50 ± 2.03）kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery，and polymerase chain reaction-restriction fragment length polymorphisms（PCR-RFLP）were used to detect CYP11B2（rs1799998）promoter region 344（C > T）base substitution. The follow-up duration was more than 6 months，with the following parameters recorded at the last follow-up：plasma aldosterone，renin，serum potassium，and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score（PASO）for primary aldosteronism，and the specific criteria were as follows. ① Clinical complete remission：the patient's blood pressure returned to normal（< 140/90 mmHg，1 mmHg = 0.133 kPa）and all antihypertensive drugs were discontinued；②Partial clinical remission：blood pressure returns to normal，and the number or dose of antihypertensive drugs is reduced compared with before；③Clinical non-remission：blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis，followed by binary logistic forward conditional regression for multivariate analysis，and a variety of machine learning algorithms such as random forest，logistic regression，support vector machine and gradient lifter were integrated，and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model，and the predictive performance of the model was evaluated by using receiver operating characteristic（ROC）curve，calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2（344C > T）（rs1799998）as follows：CC type in 50 cases（43.1%），CT type in 46 cases（39.7%），and TT type in 20 cases（17.3%）. There were 74 cases in the complete remission group and 42 cases in the incomplete remission group，and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index［（24.27 ± 2.90）kg/m 2 vs.（26.98 ± 3.17）kg/m 2， P<0.001］，preoperative hypertension grade（grade 1/2/3：29/29/16 cases vs. 9/13/20 cases， P = 0.012），preoperative antihypertensive drugs（0/1/≥ 2：25/32/17 cases vs. 7/15/20 cases， P = 0.016），and CYP11B2（344C > T）（CC/TT + CT：39/35 cases vs. 11/31 cases， P = 0.006）were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs［≥ 2： OR = 5.26（95% CI 1.12?24.61， P = 0.016；1： OR = 4.55（95% CI 1.23?22.47）， P = 0.025］was the strongest independent predictor，followed by CYP11B2（344C > T）［ OR = 4.02（95% CI 1.16?13.82）， P = 0.028］and BMI［ OR = 3.96（95% CI 2.26?6.92）， P < 0.001］. Comparing the receiver operating feature（ROC）curves of the four types of machine learning models，the best model was the support vector machine model with an area under the curve（AUC）of 0.88（95% CI 0.82?0.95），followed by the gradient elevator model of 0.83（95% CI 0.76?0.91），the logistic regression model of 0.78（95% CI 0.68?0.88），and the random forest model of 0.77（95% CI 0.68?0.86）. The optimal threshold of the Yoden index of the support vector machine model was 0.588，with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms，BMI，and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients，providing new evidence for personalized treatment strategies

Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Objective:To evaluate the effect of buccal acupuncture on analgesia after tonsilloadenoidectomy in pediatric patients.Methods:This was a randomized controlled study. One hundred and twenty-six American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ pediatric patients, aged 3-12 yr, weighing 12-34 kg, with body mass index <30 kg/m 2, undergoing elective tonsilloadenoidectomy with general anesthesia, were divided into 2 groups ( n=63 each) by the random number table method: buccal acupuncture group (group B) and control group (group C). All pediatric patients received the same anesthesia induction and intraoperative anesthesia maintenance. The concentration of sevoflurane was adjusted to keep the fluctuation amplitude of vital sign parameters within 20% of the baseline value. After surgery, the drug was immediately stopped and the children were transferred to the postanesthesia care unit for resuscitation under general anesthesia. In group B, the bilateral neck points, upper neck points, hologram points on the head and Zhongjiao points were selected before removal of the tracheal catheter, and disposable acupuncture needles were inserted directly into the acupoints and remained for 20-30 min. Group C received no buccal acupuncture. The pain Assessment Scale (FLACC) was used to assess the severity of postoperative pain. The postoperative agitation score was evaluated by Aono four-point rating method to evaluate the occurrence of agitation. The effective pressing times of patient-controlled analgesia, rescue analgesia and occurrence of nausea and vomiting within 48 h after operation were recorded. The occurrence of bleeding, infection and broken needle at acupuncture sites was recorded. Results:Compared with group C, the effective pressing times of patient-controlled analgesia and incidence of nausea and vomiting were significantly decreased in group B ( P<0.05). There was no significant difference in the rate of rescue analgesia and incidence of postoperative agitation between the two groups ( P>0.05). No infection or broken needle was found at acupuncture sites after buccal acupuncture, only 2 cases had slight bleeding at the puncture site, and there was no abnormality after pressing in group B. Conclusions:Buccal acupuncture can enhance the analgesic effect after tonsilloadenoidectomy in pediatric patients.

Objective:To investigate molecular mechanisms underlying the inflammatory response induced by Cutibacterium acnes ( C. acnes) biofilms in human primary keratinocytes. Methods:A C. acnes biofilm model was established in vitro, and confocal fluorescence microscopy was performed to examine its three-dimensional structure. The cultured human primary keratinocytes were divided into 3 groups: a dimethyl sulfoxide (DMSO) control group (treated with 0.01% DMSO alone), a C. acnes suspension group (co-incubated with C. acnes suspensions), and a C. acnes biofilm group (co-incubated with C. acnes biofilms). Real-time fluorescence-based quantitative PCR (RT-qPCR) was performed to determine the relative mRNA expression of interleukin (IL) -6, IL-8, and tumor necrosis factor (TNF) -α in the groups after 6-hour culture, enzyme-linked immunosorbent assay to detect the free protein levels of IL-6, IL-8, and TNF-α in the groups after 24-hour culture, and Western blot analysis to determine the protein expression of Toll-like receptor 2 (TLR2) in keratinocytes. In addition, some human primary keratinocytes were pretreated with key molecular blockers targeting the TLR2/mitogen-activated protein kinase (MAPK) /nuclear factor (NF) -κB signaling pathway (C29, ST2825, BAY11-7082, SB203580, U0126-EtOH), and then co-incubated with C. acnes biofilms; the DMSO control group and the C. acnes biofilm group receiving no pretreatment were simultaneously set as negative and positive controls, respectively. The mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were then detected in the above groups. One-way analysis of variance was used for comparisons among multiple groups, and the Bonferroni method was used for multiple comparisons. Results:Confocal fluorescence microscopy demonstrated a three-dimensional C. acnes biofilm structure resembling a lawn, and the biofilm grew well. RT-qPCR and ELISA showed significant differences in the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α among the C. acnes biofilm group, C. acnes suspension group and DMSO control group (mRNA: F = 89.70, 312.17, 46.09, respectively, all P < 0.001; free protein: F = 886.12, 634.25, 307.01, respectively, all P < 0.001) ; in detail, the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were significantly higher in the C. acnes biofilm group than in the C. acnes suspension group and DMSO control group (all P < 0.001) ; the C. acnes suspension group showed significantly increased expression levels of IL-6 mRNA and TNF-α free protein compared with the DMSO control group ( P < 0.001, = 0.003, respectively), while there were no significant differences in the expression of IL-6 free protein, TNF-α mRNA, or IL-8 mRNA and free protein between the 2 groups (all P > 0.05). Western blot analysis showed that the TLR2 protein expression was significantly higher in the C. acnes suspension group and C. acnes biofilm group than in the DMSO control group. After the pretreatment with molecular blockers targeting the MAPK/NF-κB signaling pathway and co-incubation with C. acnes biofilms, the mRNA and free protein expression levels of IL-6, IL-8 and TNF-α were all significantly lower in the C29 group, ST2825 group, BAY11-7082 group, SB203580 group, U0126-EtOH group, as well as in the DMSO control group compared with the C. acnes biofilm group (all P < 0.05) . Conclusion:The C. acnes biofilms exhibited a strong ability to induce inflammatory responses in human keratinocytes, possibly through the activation of the TLR2/MAPK/NF-κB signaling pathway.