Objective To investigate the effects of ROCK-siRNA transfection on endothelial cell senescence and endothelial microparticles (EMPs) induced by angiotensin II (Ang II). Methods Human umbilical vein endothelial cells (HUVECs) were treated with Ang II (1.0 μmo/L) to induce cellular senescence models, followed by transfection with ROCK-siRNA. The cells were divided into four groups: control group, model group, negative transfection control group (Ang II combined with NC-siRNA), and ROCK-siRNA transfection group (Ang II combined with ROCK-siRNA). Cellular senescence was assessed by SA-β-Gal staining. EMP levels in cell supernatants and intracellular reactive oxygen species (ROS) levels were assessed using flow cytometry. The expression levels of silenced information regulator 1(SIRT1) and p53 protein in each group were analyzed by Western blotting. Results Following ROCK-siRNA transfection, the number of senescent cells induced by Ang II was significantly reduced, accompanied by decreased CD31⁺ EMP levels and suppressed intracellular ROS levels. Meanwhile, the expression levels of SIRT1 were up-regulated, while the expression levels of p53 were down-regulated. Conclusion Silencing ROCK expression suppresses EMP release, reduces ROS generation, regulates the expression of SIRT1 and p53, and ultimately attenuates Ang II-induced endothelial cell senescence.
Humans ; Angiotensin II/pharmacology* ; Cellular Senescence/genetics* ; Human Umbilical Vein Endothelial Cells/cytology* ; RNA, Small Interfering/metabolism* ; Reactive Oxygen Species/metabolism* ; Sirtuin 1/genetics* ; Transfection ; Tumor Suppressor Protein p53/genetics* ; Cell-Derived Microparticles/drug effects* ; rho-Associated Kinases/metabolism* ; Endothelial Cells/metabolism* ; Cells, Cultured

Objective:To investigate the expression characteristics of plasma proteins in patients with acute diquat poisoning by proteomic technology, and to screen the targets and biomarkers related to the diagnosis, treatment and prognosis of acute diquat poisoning.Methods:This study was a retrospective case-control study. Patients with diquat poisoning admitted to Nanjing University Affiliated Drum Tower Hospital from August 2022 to December 2023 were enrolled. Healthy volunteers matched for age and gender during the same period were selected as the control group. The general clinical data and plasma samples were collected, and qualitative and quantitative analyses of plasma proteins were performed by proteomic technology based on nanoliter liquid chromatography tandem quadrupole time-of-flight mass spectrometry. Mann-Whitney U test was used to compare differences of plasma protein expression between the two groups. Results:A total of 8 diquat poisoning patients and 7 healthy individuals were included in this study, and 491 plasma proteins were identified. Compared with control group, 76 differentially expressed proteins were obtained from the poisoned group, including 35 up-regulated proteins and 41 down-regulated proteins. GO analysis showed that the differentially expressed proteins were mainly associated with oxygen transporter activity, receptor binding, and hyaluronic acid-binding endopeptidase inhibitor activity. KEGG pathway enrichment analysis revealed that differentially expressed proteins were associated with complement and coagulation cascades, neutrophil extracellular trap formation, and platelet activation. In addition, the expression of adiponectin (ADIPOQ) was not only increased in poisoned patients, but also significantly higher in non-survivors than in survivors ( P<0.01). Conclusions:Multiple plasma proteins with abnormal expression may be involved in the pathological process of acute diquat poisoning. ADIPOQ may have potential application value in the diagnosis, treatment, and prognosis of acute diquat poisoning.

Objective: To construct and to validate a nomogram prediction model based on Lipoprotein-associated phospholipase A2(Lp-PLA2) and Lipoprotein(a) [LP(a) ]for predicting the risk of major adverse cardiovascular events(MACE) in patients with coronary heart disease(CHD). Methods: A retrospective analysis was conducted on the clinical data of 442 patients with coronary heart disease(CHD). Among them,411 patients who completed follow-up were randomly divided into a training set(288 cases) and a validation set(123 cases) at a 7 ∶ 3 ratio.Independent risk factors for major adverse cardiovascular events(MACE) in CHD patients were screened through Lasso regression analysis and Cox regression analysis,and a nomogram prediction model was constructed. The predictive performance of the model was evaluated using time-dependent receiver operating characteristic curves(ROC),calibration curves,and decision curve analysis. Results: Variables were screened through Lasso regression and Cox regression analysis. The final model included nine independent predictors,namely age,smoking history,clinical phenotype of CHD,the number of coronary artery lesions,Gensini score,BNP,Lp-PLA2,LP(a), and the history of statin use. The area under the ROC curve in the training set was 0. 897,0. 885,and 0. 909 at 1,2,and 3 years,respectively; The area under the ROC curve in the validation set was 0. 885,0. 881,and 0. 923 at 1,2,and 3 years,respectively. These results demonstrated that the model had excellent discriminatory power. The calibration curves and decision curves demonstrated that the model had high clinical practicality in predicting the occurrence of MACE in CHD patients. Conclusion The nomogram prediction model based on LP-PLA2,LP(a)and other risk factors provides an effective tool for the prognosis assessment of CHD patients,facilitating the early identification of high-risk patients and enabling individualized intervention.

Objective:To investigate the relationship between carotid intraplaque hemorrhage (IPH) in CT angiography(CTA) and perivascular fat density (PFD).Methods:This study retrospectively analyzed 103 patients with carotid atherosclerosis, including 209 carotid plaques, who received carotid CTA and high-resolution magnetic resonance angiography in Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 2018 to July 2023. According to the evaluation of IPH by MRI, the plaques were divided into IPH group ( n=85) and non IPH group ( n=124). CTA was used to evaluate the plaque characteristics (calcification, lumen stenosis, maximum plaque thickness and plaque density) and PFD. Univariate and multivariate logistic analysis was used to evaluate the relationship between PFD and IPH. The diagnostic efficacy of multiple features was performed by receiver operating characteristic curve analysis. Results:Finally, 103 patients with 209 carotid plaques were collected. Multivariate logistic regression analysis showed that PFD ( OR=1.183, 95% CI 1.104-1.267, P<0.001) still showed a significant association with IPH after adjusting for age, hyperlipidemia, degree of luminal stenosis, plaque calcification, maximum plaque thickness, and plaque density. The area under the curve values, sensitivity, specificity, and accuracy of combined PFD, maximum plaque thickness, and plaque density were 0.980, 88.24%, 98.39%, and 94.26%, respectively. Conclusion:The changes of PFD in the carotid artery are closely related to the presence of carotid IPH in MRI and may become a new imaging indicator for high-risk plaque identification and cerebrovascular risk assessment.

Objective:In carbon ion treatment planning of water phantom, establish a conversion factor calculation system and conversion factor curves for organs at risk (OAR) for microdosimetric kinetic models (MKM) and local effect models (LEM), and validate them in clinical patient planning.Methods:Using a uniform spherical water phantom as the research object, relative biological effectiveness-weighted doses (RWD) for the LEM were re-calculated based on the physical dose of RayStation-MKM. The median dose within the planning target volume (PTV) of LEM and MKM was regarded as the conversion factor. The impacts of single-fraction target prescription dose, spread-out Bragg peak (SOBP) width and depth, shape, and irradiation mode on the conversion factor were assessed, and a conversion factor calculation system was established. Additionally, the accuracy of the conversion factor calculation system was validated using both water phantoms and clinical patient cases. The conversion factor curves for OAR were computed based on clinical patient treatment plans.Results:The primary influencing factors for the conversion factors were the single-fraction prescription dose, target SOBP width and depth. The conversion factors were increased with the increase of SOBP width and target depth, whereas decreased with the increase of the single-fraction prescription dose. Under single-field irradiation, a conversion factor calculation system was established based on above 3 parameters. For the plans of 9 patients, the average difference between the calculated results and the conversion factor calculation system was 0.340% ± 0.203%, and the average difference in the conversion curves for OAR was 2.650% ± 2.399%.Conclusion:A dose conversion factor calculation system and conversion factor curves for OAR for carbon ion radiotherapy are established for MKM and LEM, and their accuracy meets the requirements for use in clinical patient treatment plans.

[Objectives]To investigate the effect of METTL3 on the malignant biological behaviors of uveal melanoma cells and to verify whether this effect is related to m6A methylation.[Methods]Uveal melanoma cell models with METTL3 knockdown,overexpression,and point mutations at m6A-related catalytic sites were constructed via lentivirus transfection.Transwell assays were used to assess cell migration and invasion;CCK8 assays were used to measure cell proliferation and flow cytometry was used to analyze apoptosis and cycle changes.[Results]The proliferation,migration and invasion abilities of C918 and MUM-2B cells with METTL3 knockdown were significantly decreased(P<0.001),while the apoptosis rate was increased.The proportion of cells in G1 phase significantly increased,whereas the proportion in the S phase significantly decreased.Cells overexpressing METTL3 showed significantly enhanced proliferation,migration and invasion abilities(P<0.001),along with a decreased apoptosis rate.In C918 cells,the proportion of cells in G1 phase decreased significantly,while the proportion in S phase increased significantly.The cell cycle distribution of MUM-2B cells did not change remarkably.Following point mutation of m6A-related catalytic sites,cell proliferation,migration and invasion decreased,and the apoptosis rate increased.In MUM-2B cells,the percentage of cells in G1 phase significantly increased;the percentage in S phase significantly decreased and the percentage in G2 phase slightly decreased.In C918 cells,the percentage of G1 phase cells significantly increased,with no significant changes in the proportions of S and G2 phases.[Conclusions]The proliferation,invasion and metastasis of uveal melanoma cells were positively correlated with the expression of METTL3,while the apoptosis rate was negatively correlated.Changes in METTL3 levels differentially affect the cell cycles in different cell lines.The effects of METTL3 on the proliferation,migration and invasion of uveal melanoma cells are related to m6A methylation modification.

Objective:To evaluate the clinical significance of morular metaplasia (MM) in fertility-preserving treatment for young patients with endometrial hyperplasia and grade 1 endometrial endometrioid carcinoma.Methods:Clinical data was retrospectively collected from patients diagnosed with endometrial hyperplasia or grade 1 endometrial endometrioid carcinoma under 40 years old who underwent progestin-based fertility-sparing treatmentat in Tianjin Medical University General Hospital between January 2018 and November 2022.Patients were divided into the MM group (37 cases) and the non-MM group (63 cases) based on pathological findings. Clinical characteristics, hysteroscopic features, treatment efficacy and fertility outcomes were compared between the two groups. The MM group was further stratified into three subgroups based on the timing of MM occurrence:(1) MM-Bef group ( n=10): MM was present in the initial endometrial curettage or hysteroscopic biopsy pathology before fertility-sparing treatment and disappeared after treatment; (2) MM-Sus group ( n=14): MM persisted consistently before and after therapy;(3) MM-Aft group ( n=13): MM was absent before therapy but appeared after treatment. The risk factors which had impact on the treatment outcomes of the patients were analyzed using univariate and multivariate Cox regression analysis. Results:The rate of polycystic ovary syndrome were higher in the MM group than the non-MM group [51% (19/37) vs 27% (17/63), P=0.014]. The complete response (CR) rate was significantly lower in the MM group than in the non-MM group [73% (27/37) vs 95% (60/63), P=0.006], and the median time to CR was significantly longer in the MM group (6.0 vs 5.0 months, P=0.005).Multivariate analysis identified that MM-Sus ( HR=0.355, 95% CI:0.174-0.723; P=0.004) and MM-Aft ( HR=0.314, 95% CI:0.145-0.681; P=0.003) were independent risk factors for delayed CR in fertility-sparing treatment. The patients in the MM group and non-MM group underwent hysteroscopic biopsy for 76 and 131 times. "Gravel-like change" was a more frequent hysteroscopic manifestation in the MM group than that in the non-MM group [18% (14/76) vs 2% (2/131), P<0.001]. Conclusions:Patients in the MM group have poorer treatment outcomes than patients in the non-MM group. MM-Sus and MM-Aft are risk factors for fertility-preserving treatment in young patients with endometrial hyperplasia or grade 1 endometrial endometrioid carcinoma. "Gravel-like change" is the characteristic hysteroscopic manifestations of MM.

Objective:To analyze the clinical characteristics of familial hypercholesterolemia (FH) in children and provide a basis for clinical diagnosis and individualized treatment.Methods:Case series study. Clinical data of 24 children with FH, who were admitted to the Department of Endocrinology in Capital Center for Children′s Health, Capital Medical University, from January 2018 to January 2025, were analyzed. Follow-ups were performed every 3-6 months and ended in January 2025. According to the results of genetic testing, the children were divided into homozygous familial hypercholesterolemia (HoFH) group and heterozygous familial hypercholesterolemia (HeFH) group. The blood lipid levels of different subtypes, the efficacy of different treatments, and clinical outcomes were compared by Mann-Whitney U test. Results:The 24 children were from 17 families, including 14 males and 10 females, with a diagnostic age of 5.0 (3.0, 9.5) years. Genetic testing results showed that 22 cases (92%) had LDLR gene variants and 2 cases (8%) had APOB gene variants, all of which were inherited from parents. There were 5 cases (21%) of HoFH and 19 cases (79%) of HeFH, and 4 previously unreported new loci were identified. There were 6 children (25%) presented with xanthomas, including 5 cases of HoFH and 1 case of HeFH. The level of low-density lipoprotein cholesterol (LDL-C) in the HoFH group was significantly higher than that in the HeFH group ( P<0.05). Regarding treatment, 11 children received dietary control without taking medicine, 6 were treated with statins, 3 with ezetimibe, and 3 with statins combined with ezetimibe, and 1 underwent liver transplantation. None of the children receiving only dietary control achieved the target LDL-C level (<3.49 mmol/L or a reduction of >50%), and there was no statistically significant difference in LDL-C before and after dietary control ( P=0.158). After treatment with statins and (or) ezetimibe, LDL-C decreased in 12 children ( P<0.05); among them, 6 cases (all HeFH) reached the target LDL-C level. There was no statistically difference in LDL-C levels before and after treatment with atorvastatin and ezetimibe in 5 HoFH children( P>0.05). One HoFH child had LDL-C reduced to the normal range after liver transplantation. No serious adverse reactions were observed in all children during drug treatment. In the detection of vascular-related complications among 12 HeFH children, only 1 child had a slight thickening of the bilateral carotid intima-media, while no abnormalities were found in the others. Conclusions:Xanthoma is a characteristic manifestation of FH, but its incidence is relatively low in HeFH children. Family history and genetic testing are key evidences for the diagnosis of FH. Dietary control has limited efficacy in children with FH, and drug treatment should be initiated as early as possible. LDL-C levels in HoFH children are more difficult to control, if drug treatment shows poor efficacy, liver transplantation may be a better option.

Objective To investigate the differences of condylar position in patients with temporomandibular disorders(TMD),and to further explore the effects of condylar position displacement and malocclusion factors on TMD.Methods According to the inclusion criteria,normal occlusion subjects without TMD(group A,n=10),normal occlusion subjects with TMD(group B,n=8),malocclu-sion subjects without TMD(group C,n=9)and malocclusion subjects with TMD(group D,n=20)were collected.Condylar position measurement(CPM)and Cadiax Ⅳ were used to locate the position of condyle in RP and ICP,and the displacement and direction of condyle in three-dimensional direction from RP to ICP were measured by vernier caliper.All the data were averaged three times and an-alyzed by SPSS26.0 statistical software.Results In the sagittal direction,the condylar displacement of group A and group B,group C and group D,and group A and group D had statistical differences(P＜0.01).The displacement of the right condyle in the vertical direc-tion of group D was greater than that of group A,and the difference was statistically significant(P＜0.05).The difference among other groups was not statistically significant.There was no significant difference in the lateral displacement of condyle between the four groups.In group A and group D,the largest proportion of condyle deviation was anterior up,while in group B and group C,the largest proportion of condyle deviation was posterior up.Conclusion RP-ICP condylar sagittal displacement is common in people with TMD.The condyle displacement of RP-ICP in malocclusion patients without TMD is not significantly different from that in normal occlusion patients without TMD.Patients with TMD malocclusion are more likely to have condylar displacement in sagittal direction and vertical direction than normal occlusion without TMD.

Objective To establish orthotopic and ectopic pancreatic cancer models in C57BL/6N mice with normal immune function using in vivo imaging technology for visual characterization.Methods Orthotopic and ectopic pancreatic cancer models were established in Kunming mice by injecting a small volume of cell suspension containing firefly luciferase-expressing Panc02-luciferase pancreatic cancer cells into the head of the pancreas or the right axillary region.In vivo imaging technology was used to optimize the modeling method and timing in Kunming mice.Subsequently,the same method was applied to C57BL/6N mice using wild-type Panc02 pancreatic cancer cells to establish orthotopic and ectopic pancreatic cancer models with intact immune function.Key parameters,including body weight,inoculation positive rate,tumor growth time,tumor volume,and pathological characteristics across different organs,were compared be-tween the orthotopic and ectopic models in C57BL/6N mice to evaluate the applicability of these models.Results Both the small animal in vivo imaging experiments in Kunming mice and the tumor growth observation in C57BL/6N mice demonstrated that the construction periods for orthotopic and ectopic pancreatic cancer models were 20 days,with survival rates exceeding 90%.The inoculation positive rates in C57BL/6N mice were 92.3%for the orthotopic model and 78.6%for the ectopic model.On day 20 post-inoculation,the tumor volumes were(117.04±109.56)mm3 for the orthotopic model and(155.68±168.73)mm3 for the ectopic model,indicating high model success rates and consistent tumor growth.HE staining revealed pathological mitotic figures and poorly differentiated tumor tissues in both models of C57BL/6N mice,with no evidence of metastasis to other organs.Conclusions Orthotopic and ectopic pancreatic cancer models in immu-nocompetent mice were successfully developed in this study,mimicking early-stage pancreatic cancer characteristics.These models pro-vide a reliable platform for screening early diagnostic biomarkers and evaluating therapeutic interventions for pancreatic cancer.