Objective:To investigate the mechanism of mitochondrial DNA (mtDNA)-reactive oxygen species (ROS)-dynamin-related protein 1 (Drp1) axis in regulating phenotypic transformation of vascular smooth muscle cells (VSMCs).Methods:(1) VSMCs were divided into a control group, a synthetic VSMCs group, and a Drp1 siRNA+synthetic VSMCs group; cells in the Drp1 siRNA+synthetic VSMCs group were transfected with 50 nmol/L Drp1 siRNA for 48 h; cells in the latter two groups were treated with 20 ng/mL platelet-derived growth factor (PDGF)-BB, while cells in the control group were treated with an equal volume of solvent. After another 24 h of culture, Drp1 expression in VSMCs, and mitochondrial Drp1 and mitofusin 2 (Mfn2) expressions were detected by Western blotting, and changes in mitochondrial morphology were detected by mitochondrial fluorescent staining. (2) VSMCs were divided into a control group, a synthetic VSMCs group, and a mitochondrial fission inhibitor 1 (Mdivi-1)+synthetic VSMCs group; cells in the Mdivi-1+synthetic VSMCs group were pretreated with 50 μmol/L Mdivi-1 for 2 h; and cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 hours of continued culture, expressions of α-smooth muscle actin (α-SMA), smooth muscle protein 22-α (SM22-α), proliferating cell nuclear antigen (PCNA), and Cyclin D1 were detected by Western blotting; invasion and migration abilities of VSMCs were detected by Transwell assay and scratch wound healing assay, respectively. (3) VSMCs were divided into a control group, a synthetic VSMCs group, and a N-acetylcysteine (NAC)+synthetic VSMCs group; cells in the NAC+synthetic VSMCs group were pretreated with 5 mmol/L NAC for 1 h; cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 h of continued culture, expressions of Drp1, phosphorylated (p)-Drp1, α-SMA, SM22-α, PCNA, and Cyclin D1 were detected by Western blotting; changes in mitochondrial morphology were detected by mitochondrial fluorescent staining; intracellular ROS level was detected by 2', 7' -dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe; cell invasion and migration abilities were detected by Transwell assay and scratch wound healing assay, respectively. (4) VSMCs were divided into a control group, a synthetic VSMCs group, and a 5-Aza-2'-deoxycytidine (5-Aza-dC)+synthetic VSMCs group; cells in the 5-Aza-dC+synthetic VSMCs group were pretreated with 2 μmol/L 5-Aza-dC for 1 h; and then, cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 h of continued culture, agarose gel electrophoresis was used to analyze the methylation degree in the mitochondrial D-loop region; intracellular ROS level was detected using DCFH-DA fluorescent probe; expressions of mitochondrial DNMT1, α-SMA, SM22-α, PCNA, and Cyclin D1 were detected by Western blotting; invasion and migration abilities were detected by Transwell assay and scratch wound healing assay, respectively.Results:(1) Compared with the control group and synthetic VSMCs group, the Drp1 siRNA+synthetic VSMCs group had significantly decreased Drp1 protein expression ( P<0.05). Compared with the control group, the synthetic VSMCs group had significantly increased Drp1 protein expression and decreased Mfn2 protein expression in the mitochondria ( P<0.05); compared with the synthetic VSMCs group, the Drp1 siRNA+synthetic VSMCs group had statistically decreased Drp1 protein expression and increased Mfn2 protein expression in the mitochondria ( P<0.05). Results of mitochondrial fluorescent staining showed that mitochondria in the control group were with filamentous structure, while mitochondrial fission in the synthetic VSMCs group was enhanced, and morphology of mitochondria in the Drp1 siRNA+synthetic VSMCs group tended to be continuous and complete. (2) Compared with the control group, the synthetic VSMCs group had statistically decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the Mdivi-1+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster cell scratch healing; compared with the synthetic VSMCs group, the Mdivi-1+synthetic VSMCs group had smaller number of migrating cells and slower cell scratch healing. (3) Compared with the control group (1.10±0.02), the synthetic VSMCs group (1.53±0.02) had significantly increased p-Drp1 protein expression ( P<0.05). Compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group (0.90±0.02) had statistically decreased p-Drp1 protein expression ( P<0.05). Results of mitochondrial fluorescent staining showed that mitochondria in cells of the control group were in a filamentous structure, while mitochondrial fission in cells of the synthetic VSMCs group was enhanced, and morphology of mitochondria in the NAC+synthetic VSMCs group tended to be continuous and complete. Results of DCFH-DA fluorescent probe showed that ROS level in the synthetic VSMCs group was higher than that in the control group, and ROS level in the NAC+synthetic VSMCs group was lower than that in the synthetic VSMCs group. Compared with the control group, the synthetic VSMCs group had significantly decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster cell scratch healing; compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group had smaller number of migrating cells and slower cell scratch healing. (4) Results of agarose gel electrophoresis showed that compared with the control group, the synthetic VSMCs group had significantly increased methylation rate in the mitochondrial D-loop region ( P<0.05); compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had statistically decreased methylation rate in the mitochondrial D-loop region ( P<0.05). Compared with the control group, the synthetic VSMCs group had statistically increased mitochondrial DNMT1 protein expression (1.03±0.03 vs. 0.55±0.03, P<0.05); and compared with the synthetic VSMCs group, the the 5-Aza-dC+synthetic VSMCs group (0.62±0.03) had significantly decreased mitochondrial DNMT1 protein expression ( P<0.05). Results of DCFH-DA fluorescent probe showed that ROS level in the synthetic VSMCs group was higher than that in the control group; ROS level in the 5-Aza-dC+synthetic VSMCs group was lower than that in the synthetic VSMCs group. Compared with the control group, the synthetic VSMCs group had significantly decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster scratch healing. Compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had smaller number of migrating cells and slower scratch healing. Conclusion:The mtDNA-ROS-Drp1 axis may regulate the phenotypic transformation of VSMCs by modulating mitochondrial epigenetic modifications.

Objective To explore the differences in neural activity between patients with anorexia nervosa(AN)and healthy controls(HC),as well as the association between these differences and symptoms in AN patients,using activation likelihood estimation(ALE)meta-analysis.Methods The literature search covered the period from 2000 to March 2025.From a pool of 588 identified studies,4 studies focusing on the neural activity differences between AN patients and HC were selected for inclusion.These studies comprised 106 participants and 21 sets of coordinates.The ALE meta-analysis method was employed,and the GingerALE software was used to systematically analyze the reported brain region changes and their peak coordinates,aiming to investigate the differences in brain functional activity between AN patients and HC.Results Compared to the HC group,AN group showed significantly enhanced activation in the left parahippocampal gyrus/amygdala(ALE value=0.39×10-2),right parahippocampal gyrus/amygdala(ALE value=0.39×10-2)and suboccipital gyrus(ALE value=0.39×10-2),along with a significant reduction in activation in Brodmann area 17(ALE value=0.61×10-2)(P<0.01,FWE corrected).Conclusion Key brain regions in AN patients including the parahippocampal gyrus,inferior occipital gyrus,and amygdala demonstrate significant functional activation abnormalities.

BACKGROUND:Cervical rotation-traction manipulation is effective and safe in the treatment of cervical spondylotic radiculopathy,and has been widely used in clinical work.However,its effects on the biomechanics of cervical vertebra and intervertebral disc and the area of intervertebral foramen have not been systematically clarified. OBJECTIVE:Based on the finite element analysis technique,a relevant research and analysis were carried out to provide digital evidence for the mechanism of effect of cervical rotation-traction manipulation in the treatment of cervical spondylotic radiculopathy. METHODS:The CT image data of a volunteer with no neck diseases were selected as the finite element model material at its left-handed physiological limit position.The initial construction of the finite element model was completed by Mimics 19.0 software,Geomagic Studio 2013 software,Hypermash 14.0 software,and ANSYS Workbench 2020 R2 software,respectively.Based on the literature,the grid division of cervical structure and the assignment of elastic modulus and elastic coefficient were completed.Based on the previous work of the team,the mechanical effects of cervical rotation-traction manipulation were simulated on the model.Effects of cervical rotation-traction manipulation on the mechanical parameters of each vertebral body and intervertebral disc in C3-T1 segment and on the cervical lateral foramen area were analyzed. RESULTS AND CONCLUSION:(1)During cervical rotation-traction manipulation,the stress of bone structure was significantly higher than that of soft tissue such as intervertebral disc.(2)When operating the technique,the stress at the top of each cervical vertebra was higher,the stress at the bottom was lower,and the stress at the facet joint and transverse process was lower.The stress at the top of the intervertebral disc was lower,the stress at the bottom was higher,but the highest point of the intervertebral disc stress was outside the top.(3)In addition,after loading the lifting force,the projected area of the C6/C7 intervertebral foramen increased significantly compared with that before loading.(4)It is indicated that the cervical rotation-traction manipulation has the mechanical characteristics of changing the stress structure of the cervical spine itself,and can expand the C6/7 intervertebral cervical foramen area on the opposite side of the patient's cervical rotation,so as to achieve the purpose of treating cervical spondylotic radiculopathy.

Objective: o develop an onset risk prediction nomogram for patients with homocysteine-type (H-type) hypertension (HTH) based on pulse diagram parameters to assist early clinical prediction and diagnosis of HTH. Methods: Patients diagnosed with essential hypertension and admitted to Shanghai Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai Hospital of Traditional Chinese Medicine, and Shanghai Hospital of Integrated Traditional Chinese and Western Medicine from July 6th 2020 to June 16th 2021, and from August 11th 2023 to January 22nd 2024, were enrolled in this retrospective research. The baselines and clinical biochemical indicators of patients were collected. The SMART-I TCM pulse instrument was applied to gather pulse diagram parameters. Multivariate logistic regression was adopted to analyze the risk factors for HTH. RStudio was employed to construct the nomogram model, receiver operating characteristic (ROC) curve, and calibration curve (bootstrap self-sampling 200 times), and clinical decision curve were drawn to evaluate the model’s discrimination and clinical effectiveness. Results: A total of 168 hospitalized patients with essential hypertension were selected and divided into non-HTH group (n = 29) and HTH group (n = 139). Compared with non-HTH group, HTH group had a lower body mass index (BMI), and higher proportions of male patients and drinkers (P < 0.05). The ventricular wall thickening (VWT) could not be determined. The proportions of left common carotid intima-media wall thickness (LCCIMWT) and serum creatinine (SCR) were higher in HTH group (P < 0.05). The pulse diagram parameter As was significantly higher, and H4/H1 and T1/T were lower in HTH group (P < 0.05). Gender, alcohol consumption, serum creatinine, and the pulse diagram parameter H4/H1 were identified as independent risk factors for HTH (P < 0.05). The nomogram’s area under the ROC curve (AUC) was 0.795 [95% confidence interval (CI): (0.706 6, 0.882 8)], with a specificity of 0.724 and sensitivity of 0.799. After 200 times repeated bootstrap self-samplings, the calibration curve showed that the simulated curve fits well with the actual curve (x2 = 9.5002, P = 0.301 9). The clinical decision curve indicated that the nomogram’s applicability was optimal when the threshold for predicting HTH was between 0.38 and 1.00. Conclusion The nomogram model could be valuable for predicting the onset risk of HTH and pulse diagram parameters can facilitate early screening and prevention of HTH.

Objective To explore the differences in neural activity between patients with anorexia nervosa(AN)and healthy controls(HC),as well as the association between these differences and symptoms in AN patients,using activation likelihood estimation(ALE)meta-analysis.Methods The literature search covered the period from 2000 to March 2025.From a pool of 588 identified studies,4 studies focusing on the neural activity differences between AN patients and HC were selected for inclusion.These studies comprised 106 participants and 21 sets of coordinates.The ALE meta-analysis method was employed,and the GingerALE software was used to systematically analyze the reported brain region changes and their peak coordinates,aiming to investigate the differences in brain functional activity between AN patients and HC.Results Compared to the HC group,AN group showed significantly enhanced activation in the left parahippocampal gyrus/amygdala(ALE value=0.39×10-2),right parahippocampal gyrus/amygdala(ALE value=0.39×10-2)and suboccipital gyrus(ALE value=0.39×10-2),along with a significant reduction in activation in Brodmann area 17(ALE value=0.61×10-2)(P<0.01,FWE corrected).Conclusion Key brain regions in AN patients including the parahippocampal gyrus,inferior occipital gyrus,and amygdala demonstrate significant functional activation abnormalities.

Objective:To study the resistance of Aedes albopictus to commonly used insecticides in Hangzhou, and to investigate the polymorphism distribution characteristics of voltage-gated sodium channel (VGSC) and acetylcholinesterase (AchE) 1 (Ace-1) genes in Aedes albopictus in the region, providing a scientific basis for control of Aedes albopictus in Hangzhou. Methods:From May to June 2023, Aedes albopictus larvae and pupae were collected from residential areas across divided different urban areas by directions in Hangzhou. They were reared in the laboratory, and their resistance to malathion, propoxur, pyrethroid insecticides (permethrin, deltamethrin, and beta-cypermethrin) was evaluated using the World Health Organization-recommended adult mosquito contact tube method. The 24 h mortality rates of adult mosquito of 98% - 100% was susceptible, 80% - < 98% was suspected resistance, < 80% was resistant. Genomic DNA was extracted from adult Aedes albopictus, and mutations in the VGSC and Ace-1 genes were detected through PCR amplification and Sanger sequencing. Results:The 24 h mortality rates of adult Aedes albopictus exposed to 0.5% malathion, 0.05% propoxur, 0.4% permethrin, 0.03% deltamethrin, and 0.08% beta-cypermethrin in Hangzhou were 100% (75/75), 96.00% (72/75), 41.43% (29/70), 17.33% (13/75), and 32.86% (23/70), respectively. A total of 146 surviving Aedes albopictus were tested for VGSC gene mutations at diagnostic doses of pyrethroid insecticides. Among them, 6 Aedes albopictus (4.11%) had a gene mutation at position V1016, and 146 Aedes albopictus (100%) had a gene mutation at position F1534. A total of 63 Aedes albopictus that died at diagnostic doses of pyrethroid insecticides were analyzed for VGSC gene mutations. No gene mutation was detected at position V1016, while 63 Aedes albopictus (100%) had a gene mutation at position F1534. One non synonymous mutation was detected at position V1016 of the VGSC gene, which was a heterozygous mutation of V1016V to V1016V/G. Two non synonymous mutations were detected at position F1534 of the VGSC gene, namely F1534F to F1534S or a F1534F/S heterozygous mutation. No Ace-1 gene mutation was detected in all Aedes albopictus exposed to 0.05% propoxur ( n = 75). Conclusions:The Aedes albopictus in Hangzhou has developed resistance to permethrin, deltamethrin, and beta-cypermethrin. It shows suspected resistance to propoxur and sensitivity to malathion. Moreover, a high frequency of mutations in the VGSC gene has been observed in these mosquitoes. It is crucial to enhance resistance monitoring in Aedes albopictus populations in Hangzhou and to implement the rational rotation of chemical insecticides to slow down the progress of drug resistance.

Objective:To study the resistance of Aedes albopictus to commonly used insecticides in Hangzhou, and to investigate the polymorphism distribution characteristics of voltage-gated sodium channel (VGSC) and acetylcholinesterase (AchE) 1 (Ace-1) genes in Aedes albopictus in the region, providing a scientific basis for control of Aedes albopictus in Hangzhou. Methods:From May to June 2023, Aedes albopictus larvae and pupae were collected from residential areas across divided different urban areas by directions in Hangzhou. They were reared in the laboratory, and their resistance to malathion, propoxur, pyrethroid insecticides (permethrin, deltamethrin, and beta-cypermethrin) was evaluated using the World Health Organization-recommended adult mosquito contact tube method. The 24 h mortality rates of adult mosquito of 98% - 100% was susceptible, 80% - < 98% was suspected resistance, < 80% was resistant. Genomic DNA was extracted from adult Aedes albopictus, and mutations in the VGSC and Ace-1 genes were detected through PCR amplification and Sanger sequencing. Results:The 24 h mortality rates of adult Aedes albopictus exposed to 0.5% malathion, 0.05% propoxur, 0.4% permethrin, 0.03% deltamethrin, and 0.08% beta-cypermethrin in Hangzhou were 100% (75/75), 96.00% (72/75), 41.43% (29/70), 17.33% (13/75), and 32.86% (23/70), respectively. A total of 146 surviving Aedes albopictus were tested for VGSC gene mutations at diagnostic doses of pyrethroid insecticides. Among them, 6 Aedes albopictus (4.11%) had a gene mutation at position V1016, and 146 Aedes albopictus (100%) had a gene mutation at position F1534. A total of 63 Aedes albopictus that died at diagnostic doses of pyrethroid insecticides were analyzed for VGSC gene mutations. No gene mutation was detected at position V1016, while 63 Aedes albopictus (100%) had a gene mutation at position F1534. One non synonymous mutation was detected at position V1016 of the VGSC gene, which was a heterozygous mutation of V1016V to V1016V/G. Two non synonymous mutations were detected at position F1534 of the VGSC gene, namely F1534F to F1534S or a F1534F/S heterozygous mutation. No Ace-1 gene mutation was detected in all Aedes albopictus exposed to 0.05% propoxur ( n = 75). Conclusions:The Aedes albopictus in Hangzhou has developed resistance to permethrin, deltamethrin, and beta-cypermethrin. It shows suspected resistance to propoxur and sensitivity to malathion. Moreover, a high frequency of mutations in the VGSC gene has been observed in these mosquitoes. It is crucial to enhance resistance monitoring in Aedes albopictus populations in Hangzhou and to implement the rational rotation of chemical insecticides to slow down the progress of drug resistance.

Objective:To investigate the mechanism of mitochondrial DNA (mtDNA)-reactive oxygen species (ROS)-dynamin-related protein 1 (Drp1) axis in regulating phenotypic transformation of vascular smooth muscle cells (VSMCs).Methods:(1) VSMCs were divided into a control group, a synthetic VSMCs group, and a Drp1 siRNA+synthetic VSMCs group; cells in the Drp1 siRNA+synthetic VSMCs group were transfected with 50 nmol/L Drp1 siRNA for 48 h; cells in the latter two groups were treated with 20 ng/mL platelet-derived growth factor (PDGF)-BB, while cells in the control group were treated with an equal volume of solvent. After another 24 h of culture, Drp1 expression in VSMCs, and mitochondrial Drp1 and mitofusin 2 (Mfn2) expressions were detected by Western blotting, and changes in mitochondrial morphology were detected by mitochondrial fluorescent staining. (2) VSMCs were divided into a control group, a synthetic VSMCs group, and a mitochondrial fission inhibitor 1 (Mdivi-1)+synthetic VSMCs group; cells in the Mdivi-1+synthetic VSMCs group were pretreated with 50 μmol/L Mdivi-1 for 2 h; and cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 hours of continued culture, expressions of α-smooth muscle actin (α-SMA), smooth muscle protein 22-α (SM22-α), proliferating cell nuclear antigen (PCNA), and Cyclin D1 were detected by Western blotting; invasion and migration abilities of VSMCs were detected by Transwell assay and scratch wound healing assay, respectively. (3) VSMCs were divided into a control group, a synthetic VSMCs group, and a N-acetylcysteine (NAC)+synthetic VSMCs group; cells in the NAC+synthetic VSMCs group were pretreated with 5 mmol/L NAC for 1 h; cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 h of continued culture, expressions of Drp1, phosphorylated (p)-Drp1, α-SMA, SM22-α, PCNA, and Cyclin D1 were detected by Western blotting; changes in mitochondrial morphology were detected by mitochondrial fluorescent staining; intracellular ROS level was detected by 2', 7' -dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe; cell invasion and migration abilities were detected by Transwell assay and scratch wound healing assay, respectively. (4) VSMCs were divided into a control group, a synthetic VSMCs group, and a 5-Aza-2'-deoxycytidine (5-Aza-dC)+synthetic VSMCs group; cells in the 5-Aza-dC+synthetic VSMCs group were pretreated with 2 μmol/L 5-Aza-dC for 1 h; and then, cells in the latter two groups were treated with 20 ng/mL PDGF-BB, while cells in the control group were treated with an equal volume of solvent. After 24 h of continued culture, agarose gel electrophoresis was used to analyze the methylation degree in the mitochondrial D-loop region; intracellular ROS level was detected using DCFH-DA fluorescent probe; expressions of mitochondrial DNMT1, α-SMA, SM22-α, PCNA, and Cyclin D1 were detected by Western blotting; invasion and migration abilities were detected by Transwell assay and scratch wound healing assay, respectively.Results:(1) Compared with the control group and synthetic VSMCs group, the Drp1 siRNA+synthetic VSMCs group had significantly decreased Drp1 protein expression ( P<0.05). Compared with the control group, the synthetic VSMCs group had significantly increased Drp1 protein expression and decreased Mfn2 protein expression in the mitochondria ( P<0.05); compared with the synthetic VSMCs group, the Drp1 siRNA+synthetic VSMCs group had statistically decreased Drp1 protein expression and increased Mfn2 protein expression in the mitochondria ( P<0.05). Results of mitochondrial fluorescent staining showed that mitochondria in the control group were with filamentous structure, while mitochondrial fission in the synthetic VSMCs group was enhanced, and morphology of mitochondria in the Drp1 siRNA+synthetic VSMCs group tended to be continuous and complete. (2) Compared with the control group, the synthetic VSMCs group had statistically decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the Mdivi-1+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster cell scratch healing; compared with the synthetic VSMCs group, the Mdivi-1+synthetic VSMCs group had smaller number of migrating cells and slower cell scratch healing. (3) Compared with the control group (1.10±0.02), the synthetic VSMCs group (1.53±0.02) had significantly increased p-Drp1 protein expression ( P<0.05). Compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group (0.90±0.02) had statistically decreased p-Drp1 protein expression ( P<0.05). Results of mitochondrial fluorescent staining showed that mitochondria in cells of the control group were in a filamentous structure, while mitochondrial fission in cells of the synthetic VSMCs group was enhanced, and morphology of mitochondria in the NAC+synthetic VSMCs group tended to be continuous and complete. Results of DCFH-DA fluorescent probe showed that ROS level in the synthetic VSMCs group was higher than that in the control group, and ROS level in the NAC+synthetic VSMCs group was lower than that in the synthetic VSMCs group. Compared with the control group, the synthetic VSMCs group had significantly decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster cell scratch healing; compared with the synthetic VSMCs group, the NAC+synthetic VSMCs group had smaller number of migrating cells and slower cell scratch healing. (4) Results of agarose gel electrophoresis showed that compared with the control group, the synthetic VSMCs group had significantly increased methylation rate in the mitochondrial D-loop region ( P<0.05); compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had statistically decreased methylation rate in the mitochondrial D-loop region ( P<0.05). Compared with the control group, the synthetic VSMCs group had statistically increased mitochondrial DNMT1 protein expression (1.03±0.03 vs. 0.55±0.03, P<0.05); and compared with the synthetic VSMCs group, the the 5-Aza-dC+synthetic VSMCs group (0.62±0.03) had significantly decreased mitochondrial DNMT1 protein expression ( P<0.05). Results of DCFH-DA fluorescent probe showed that ROS level in the synthetic VSMCs group was higher than that in the control group; ROS level in the 5-Aza-dC+synthetic VSMCs group was lower than that in the synthetic VSMCs group. Compared with the control group, the synthetic VSMCs group had significantly decreased α-SMA and SM22-α protein expressions and increased PCNA and Cyclin D1 protein expressions ( P<0.05). Compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had significantly increased α-SMA and SM22-α protein expressions and decreased PCNA and Cyclin D1 protein expressions ( P<0.05). Results of Transwell and scratch wound healing assays showed that compared with the control group, the synthetic VSMCs group had larger number of migrating cells and faster scratch healing. Compared with the synthetic VSMCs group, the 5-Aza-dC+synthetic VSMCs group had smaller number of migrating cells and slower scratch healing. Conclusion:The mtDNA-ROS-Drp1 axis may regulate the phenotypic transformation of VSMCs by modulating mitochondrial epigenetic modifications.

OBJECTIVE To optimize the management model of drugs used in investigator-initiated trial(IIT).METHODS With benchmarking analysis,based on the practical work experience of a tertiary specialized hospital in the field of IIT drug management in Shanghai,a thorough review was conducted,involving relevant laws,regulations,and academic literature to establish benchmark criteria and the evaluation standards.Starting from the initiation of IIT projects,a detailed comparative analysis of key processes was carried out,such as the receipt,storage,distribution,use and recycling of drugs for trial.The deficiencies in the current management of IIT drugs were reviewed in detail and a series of optimization suggestions were put forward.RESULTS It was found that the authorized records of drug management were missing,the training before project implementation was insufficient,and the records of receipt and acceptance of IIT drugs were incomplete.In light of these existing problems,improvement measures were put forward,including strengthening the training of drug administrators and stipulating that only drug administrators with pharmacist qualifications be eligible to inspect and accept drugs,etc.The related systems were improved,and 17 key points of quality control for the management of IIT drugs were developed.CONCLUSIONS A preliminary IIT drug management system for medical institutions has been established,which helps to improve the institutional framework of medical institutions in this field.

Objective:To investigate the resistance of Culex pipiens pallens to pyrethroid insecticides in Hangzhou City, and explore the mutation of knock down resistance (kdr) gene of Culex pipiens pallens, providing scientific basis for control of Culex pipiens pallens in the region. Methods:In September 2022, larvae and pupae of Culex pipiens pallens were collected from various locations at national monitoring sites in Shangcheng District and Gongshu District of Hangzhou City. They were bred in the laboratory and their resistance to three commonly used pyrethroid insecticides (permethrin, deltamethrin and beta-cypermethrin) was determined using the adult mosquito contact tube method and larval immersion method recommended by the World Health Organization. The genomic DNA was extracted from individual adult mosquito, and kdr gene mutation was detected via PCR amplification and Sanger sequencing. Results:The 24-hour mortality rates of adult mosquitoes of Culex pipiens pallens exposed to 0.25% permethrin, 0.025% deltamethrin, and 0.025% beta-cypermethrin were 20.00% (15/75), 17.33% (13/75), and 18.67% (14/75), respectively, indicating resistance. The resistance ratios of Culex pipiens pallens larvae to permethrin, deltamethrin, and beta-cypermethrin were 27.08, 341.00, and 15.88, respectively. Among 183 adult mosquitoes that survived under the diagnostic doses of pyrethroid insecticides, 180 mosquitoes (98.36%) had mutation at the L1014 site of the kdr gene. Among 42 adult mosquitoes that died under the diagnostic doses of pyrethroid insecticides, 5 mosquitoes (11.90%) had L1014 mutation in the kdr gene. Conclusions:Culex pipiens pallens in Hangzhou City has developed varying degrees of resistance to permethrin, deltamethrin and beta-cypermethrin, with a high frequency of kdr gene mutation in resistant mosquitoes. We should strengthen the monitoring of the resistance of Culex pipiens pallens in Hangzhou City and use chemical insecticides reasonably.