AIM: To develop a novel gene-delivery therapeutic based on CRISPR/Cas9 genome editing technology capable of specifically targeting and knocking out the VEGFA gene, thereby achieving sustained suppression of VEGFA expression in retinal pigment epithelial(RPE)cells and providing a new strategy for gene therapy in retinal neovascular diseases.METHODS:Single guide RNAs targeting the human VEGFA gene for knockout were designed, and corresponding recombinant plasmids were constructed. A novel polymer(PTEE)was used to encapsulate the plasmids to prepare a PTEE-loaded anti-VEGFA plasmid(PLAP)gene delivery system. PTEE materials at concentrations of 0.1, 0.2, 0.4, 0.8, and 1.6 μg/μL were co-incubated with ARPE-19 cells, and the biocompatibility of PTEE was evaluated using the cell counting kit-8(CCK-8)assay. Recombinant plasmids expressing green fluorescent protein(GFP)were constructed. Lipofectamine 3000 and jetOPTIMUS®DNA transfection reagents were used as control groups, and PTEE nanomaterials were used as the experimental group to encapsulate the plasmids. When the cell confluence reached 80%, the formulations were transfected into ARPE-19 and 293T cells. GFP expression was observed under light microscopy, and the transfection efficiencies of each group were compared. ARPE-19 cells were induced under hypoxia, and PLAP was transfected into the cells. The expression level of VEGFA was detected by enzyme-linked immunosorbent assay(ELISA)to evaluate the efficacy of this novel gene delivery system.RESULTS: After co-incubation of ARPE-19 cells with different concentrations of PTEE for 24 h and 48 h, no significant effect on cell viability was observed in any group. The transfection efficiency of PLAP in ARPE-19 cells was higher than that in the Lipo3000 and jetOPTIMUS groups, with statistically significant differences(P<0.01). Hypoxia for 6 h significantly induced the upregulation of VEGFA mRNA expression in ARPE-19 cells, and under hypoxic conditions, the PTEE group exhibited a significant inhibitory effect on VEGFA expression(P<0.01).CONCLUSION:PLAP exhibits favorable biocompatibility and prominent VEGFA inhibitory effects in vitro, making it a potential candidate drug for gene therapy of retinal neovascular diseases.

Medical academic conferences represent one of the important carriers of scientific and technological exchanges， promoting the progress of medical science and technology and the dissemination of knowledge. However， the surge in the number of conferences has not been matched by a corresponding improvement in academic quality， leading to widespread doubts about their academic nature. The standardization of medical academic conferences urgently requires clarifying the core value standards of high-quality medical academic conferences from the perspective of academic ethics. Based on the perspective of academic ethics and employing Bourdieu’s field theory， this study systematically analyzed the functional positioning and value norms of medical academic conferences. It argued that they were committed to “pursuing truth” in knowledge production at the individual level， focused on “pursuing goodness” in communication and dissemination at the academic community level， and strove for “pursuing innovation” in innovative influence at the social level. Taking a professional annual conference as a typical case， this study further delved into the operational processes and internal mechanisms of academic annual conferences in the medical field. On this basis， optimization recommendations were proposed， including strengthening the principle of academic priority， promoting equal participation mechanisms， fostering innovative development models， and other aspects， with a view to providing a theoretical basis and practical guidance for the high-quality development of medical academic conferences.

Homo- or heterodimeric compounds that affect dimeric protein function through interaction between monomeric moieties and protein subunits can serve as valuable sources of potent and selective drug candidates. Here, we screened an in-house dimeric natural product collection, and panepocyclinol A (PecA) emerged as a selective and potent STAT3 inhibitor with profound anti-tumor efficacy. Through cross-linking C712/C718 residues in separate STAT3 monomers with two distinct Michael receptors, PecA inhibits STAT3 DNA binding affinity and transcription activity. Molecular dynamics simulation reveals the key conformation changes of STAT3 dimers upon the di-covalent binding with PecA that abolishes its DNA interactions. Furthermore, PecA exhibits high efficacy against anaplastic large T cell lymphoma in vitro and in vivo, especially those with constitutively activated STAT3 or STAT3^Y640F. In summary, our study describes a distinct and effective di-covalent modification for the dimeric compound PecA to disrupt STAT3 function.

OBJECTIVES: To investigate the antioxidative mechanism of snake venom-derived protein C activator (PCA) in mitigating vascular endothelial cell injury. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in DMEM containing 1.0 g/L D-glucose and exposed to hypoxia (1% O₂) for 6 h followed by reoxygenation for 2 h to establish a cell model of oxygen-glucose deprivation/reoxygenation (OGD/R). The cell model was treated with 2 μg/mL PCA alone or in combination with 2-ME2 (a HIF-1α inhibitor) or DMOG (a HIF-1α stabilizer), and intracellular production of reactive oxygen species (ROS) and protein expression levels of HIF-1α, BNIP3, and Beclin-1 were detected using DCFH-DA fluorescence probe, flow cytometry, and Western blotting. The OGD/R cell model was transfected with a BNIP3-specific siRNA or a scrambled control sequence prior to PCA treatment, and the changes in protein expressions of HIF-1α, BNIP3 and Beclin-1 and intracellular ROS production were examined. RESULTS: In the OGD/R cell model, PCA treatment significantly upregulated HIF-1α, BNIP3 and Beclin-1 expressions and reduced ROS production. The effects of PCA were obviously attenuated by co-treatment with 2-ME2 but augmented by treatment with DMOG (a HIF-1α stabilizer). In the cell model with BNIP3 knockdown, PCA treatment increased BNIP3 expression and decreased ROS production without causing significant changes in HIF-1α expression. Compared with HUVECs with PCA treatment only, the cells with BNIP3 knockdown prior to PCA treatment showed significantly lower Beclin-1 expression and higher ROS levels. CONCLUSIONS Snake venom PCA alleviates OGD/R-induced endothelial cell injury by upregulating HIF-1α/BNIP3 signaling to suppress ROS generation, suggesting its potential as a therapeutic agent against oxidative stress in vascular pathologies.
Humans ; Reactive Oxygen Species/metabolism* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Human Umbilical Vein Endothelial Cells/drug effects* ; Membrane Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Up-Regulation ; Cell Hypoxia ; Cells, Cultured ; Snake Venoms/chemistry* ; Beclin-1

Objective: To investigate the risk factors affecting the early diagnosis of ABO-hemolytic disease of the fetus and newborn (ABO-HDFN) in neonates with maternal-fetal blood group incompatibility, and to develop a risk prediction model and validate its predictive performance, so as to provide a reference for the early diagnosis of neonates with ABO-HDFN in primary hospitals. Methods: A total of 1 229 neonates with maternal-fetal blood group incompatibility suspected of ABO-HDFN, admitted to our hospital between between June 2021 and September 2024, were enrolled. The sample size was calculated by using the events per variable (EPV) method. The cohort was divided into a modeling group (n=860) and a validation group (n=369), and the results and clinical information of laboratory examination indicators were collected. Univariate and multivariate logistic regression analysis were used to explore the risk factors affecting the early diagnosis of ABO-HDFN in neonates with maternal-fetal blood group incompatibility. The risk prediction model was developed and internally validated by the Bootstrap method. The goodness-of-fit of the model was evaluated by the Hosmer-Lemeshow (H-L) test, and the receiver operating characteristic (ROC) curve was used to analyze the predictive performance of the model. The prediction model was validated by using the validation group data, and the predictive performance of the model was evaluated. Results: Among the 860 neonates with maternal-fetal incompatibility in the modeling group, 346 (346/860, 40.23%) were diagnosed with ABO-HDFN. Univariate and multivariate logistic regression analyses identified the following as significant risk factors for early diagnosis: the number of postnatal days at specimen collection, maternal type O blood group, parity >1, time of onset for pathologic jaundice, maternal-fetal blood group incompatibility due to A antigen, the level of total bilirubin, and the immature reticulocyte fraction (IRF). A risk prediction model was established, and the calibration degree of the model was validated by the Bootstrap internal validation method, Brier=0.143. The results of H-L test showed that χ =3.464, P=0.902. The area under the ROC curve (AUC) was 0.885. The maximum value of the Youden index was 0.611, the sensitivity was 0.832, and the specificity was 0.778. The results of the validation group showed that the area under the ROC curve was 0.863, with a sensitivity of 0.875 and specificity of 0.735. Conclusion: The risk prediction model developed based on these risk factors has good predictive performance for ABO-HDFN, facilitating early diagnosis of suspected ABO-HDFN cases by clinicians in primary hospitals.

Objective: To understand the prevalence of elevated blood pressure and its association with dietary patterns in children and adolescents in China, providing evidence for developing dietary intervention of hypertension in children and adolescents. Methods: Data were derived from the China Children s Nutrition and Health System Survey and Application Project(2019-2021). A stratified cluster random sampling method was used to include 7 933 participants from 28 survey sites in seven major regions of Northeast, North, Northwest, East, Central, South and Southwest China. Multivariate Logistic regression models were used to analyze associations between demographic characteristics, nutritional status and elevated blood pressure. Exploratory factor analysis identified dietary patterns, which were divided into three quartile groups (T3, T2, T1) based on factor scores (compliance for dietary pattern) from high to low, and multivariate Logistic regression model assessed the correlation between elevated blood pressure and dietary patterns. Results: The prevalence of elevated blood pressure was 15.4% among Chinese children aged 7-17 years. Significant differences were observed across nutritional status (reference: underweight; normal weight: OR =1.57; overweight: OR = 2.61 ; obesity: OR =3.85), urban/rural residence (reference: rural; urban: OR =0.86), and paternal education (reference: junior high school and below; bachelor degree or above: OR =0.68) ( P <0.05). The detection rates of high blood pressure in T3 group children and adolescents with four dietary patterns (staple food, animal based food, snacks, vegetables and fruits) were 15.7%, 14.6%, 16.8%, and 15.8%, respectively. After adjusting for residence, paternal education, and nutritional status, the "snack dietary pattern" (mainly candy, sugar sweetened beverages, and processed snacks) showed positive associations with elevated blood pressure in T2 ( OR =1.21) and T3 ( OR =1.19) tertiles ( P <0.05). Conclusions The snack dietary pattern is a related factor for elevated blood pressure in children and adolescents. Restricting unhealthy snack intake may promote cardiovascular health.

OBJECTIVE To establish the fingerprint of Bushen ningshen ointment， determine the contents of its major constituents， and investigate its in vitro antioxidant activity. METHODS High performance liquid chromatography （HPLC） fingerprints of 10 batches of Bushen ningshen ointment were established. Similarity evaluation and identification of common peaks were subsequently performed. The contents of 10 components such as salidroside were determined using the same HPLC method. Using the scavenging rates against 2，2′-azino bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） and 1，1-diphenyl-2- picrylhydrazyl （DPPH） radicals， as well as ferric ion reducing antioxidant power （FRAP） as indicators， the anti-oxidant activity of the ointment was evaluated； grey relational analysis and partial least squares regression were conducted using SIMCA 14.1 software to establish the spectrum-effect relationship. RESULTS The fingerprint chromatogram of 10 batches of Bushen ningshen ointment contained 24 common peaks， with similarity values all exceeding 0.96. Eleven peaks were identified as adenosine （peak 1）， salidroside （peak 4）， morroniside （peak 6）， catechin （peak 7）， paeoniflorin （peak 10）， spinosin （peak 11）， ferulic acid （peak 12）， isoquercitrin （peak 13）， E-mail：wli1743@163.com verbascoside （peak 14）， paeonol （peak 23）， and emodin （peak 24）. Content determination results showed that the average contents of salidroside， morroniside， catechin， paeoniflorin， spinosin， ferulic acid， isoquercitrin， verbascoside， paeonol， and emodin were 0.725， 1.962， 0.214， 3.395， 0.124， 0.107， 0.286， 0.019， 0.034 and 0.067 mg/g， respectively. The antioxidant potency composite index （APC） for the 10 batches ranged from 85.08% to 96.35%. Spectrum-effect relationship analysis indicated that all 24 common peaks were positively correlated with the antioxidant capacity. Seventeen peaks had variable importance in projection values ＞1， specitically peaks 2， 5， 6， 7， 9， 10， 13- 21， 23， and 24. CONCLUSIONS This study successfully established the HPLC fingerprint and content determination method for Bushen ningshen ointment. The compounds represented by the 17 common peaks such as morroniside may be the active components contributing to its antioxidant effects.

ObjectiveTo investigate the expression and prognostic significance of human epidermal growth factor receptor 2 （HER-2）， vascular endothelial growth factor-A （VEGF-A）， and programmed death-ligand 1 （PD-L1） in gallbladder cancer， and to provide a theoretical basis for subsequent research. MethodsA retrospective analysis was performed for the postoperative specimens and clinical data of 55 patients who underwent radical cholecystectomy for gallbladder cancer and had pathologically confirmed gallbladder adenocarcinoma in The Affiliated Hospital of Inner Mongolia Medical University from December 2017 to September 2019. Immunohistochemical staining was used to measure the expression levels of HER-2， VEGF-A， and PD-L1 in cancerous tissue and paracancerous tissue. The association between the expression of these three markers and clinical features were analyzed， as well as their impact on the prognosis of patients. The independent-samples t test was used for comparison of normally distributed continuous data between groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups； the chi-square test was used for comparison of categorical data between groups. The Cox regression model was used to perform the univariate and multivariate analyses of clinical factors. The Kaplan-Meier curve was used to analyze the prognosis of patients， and the Log-rank test was used for comparison. ResultsAmong the 55 patients， 15 patients （27.2%） had a high expression level of HER-2 in cancerous tissue， 23 （41.8%） had a high expression level of VEGF-A， and 18 （32.7%） had a high expression level of PD-L1， with significant differences compared with the paracancerous tissue （all P<0.05）. There was a significant difference in tumor location between the groups with different expression levels of HER-2 （P<0.05）； there were significant differences in maximum tumor diameter， degree of tumor differentiation， tumor location， N stage， peripheral organ invasion， and vascular invasion between the groups with different expression levels of VEGF-A （all P<0.05）； there were significant differences in peripheral organ invasion， vascular invasion， and disease stage between the groups with different expression levels of PD-L1 （all P<0.05）. The multivariate Cox analysis showed that preoperative CA19-9 level and HER-2 expression were independent risk factors for overall survival of patients （P<0.05）， and preoperative CA19-9 level， maximum tumor diameter， N stage， VEGF-A， and PD-L1 were independent influencing factors for progression-free survival of patients （P<0.05）. The Kaplan-Meier analysis showed that there were significant differences in overall survival and progression-free survival between the patients with high HER-2 expression and those with low HER-2 expression （P<0.05）. ConclusionHER-2， VEGF-A， and PD-L1 have an important clinical significance for patients with gallbladder cancer and are potential sites for targeted therapy.

Objective: To analyze the consumption frequency of major foods among Chinese children aged 6-17 years old, and to provide a basis for optimizing the dietary structure of children in China. Methods: Using data from the China Nutrition and Health System Survey and Application Program for Children 0-18 years old, 56 734 children aged 6-17 years old from North, Norththeast East, Central, South, Southwest and Northwest seven regions in China were selected for the study using stratified cluster random sampling from 2019 to 2021. A food frequency questionnaire was used to investigate the intake frequency of eight food groups in a month, including fresh vegetables, fresh fruits, livestock and poultry meats, aquatic products, eggs, dairy products, legumes, and cereals and potatoes. The foods were grouped according to whether they met the recommended intake criteria outlined in the Dietary Guidelines for Chinese Residents 2022. The〖KG*2〗χ2 test was used to compare the differences in the proportion of childrens intake frequency of each food group meeting the standard in different regions and age groups. Results: The proportions of Chinese children aged 6-17 years who consumed fresh vegetables and cereals and potatoes ≥3 times/d were 12.1% and 67.2%, respectively. The proportions of children who consumed fresh fruits, livestock and poultry meats, eggs and dairy products ≥1 time/d were 50.8%, 58.8%, 36.0% and 54.3%, respectively. The proportion of legumes consumed ≥4 times/week was 37.4%, and the proportion of aquatic products consumed ≥2 times/week was 39.7%. Fresh vegetables (5.5%), fresh fruits (33.1%), and dairy products (36.4%) had the lowest frequency of meeting the recommended standards in South China, and aquatic products (27.4%) and eggs (21.1%) had the lowest frequency of meeting the recommended standards in Northwest (P<0.008 3). Conclusion The overall intake frequency of fresh vegetables, fresh fruits, legumes, and dairy products are insufficient among Chinese children, with significant regional variations.

Objective To investigate the setup error in patients with spinal bone metastasis who underwent radiotherapy under the guidance of kilovoltage cone-beam CT (KV-CBCT). Methods A total of 118 patients with spinal metastasis who underwent radiotherapy, including 17 cases of cervical spine, 62 cases of thoracic spine, and 39 cases of lumbar spine, were collected. KV-CBCT scans were performed using the linear accelerators from Elekta and Varian’s EDGE system. CBCT images were registered with reference CT images in the bone window mode. A total of 973 data were collected, and 3D linear errors were recorded. Results The patients with spinal bone metastasis were grouped by site, height, weight, and BMI. The P value of the patients grouped only by site was P<0.05, which was statistically significant. Conclusion When grouped by site in the 3D direction, the positioning effect of cervical spine is better than that of thoracic and lumbar spine. The positioning effect of the thoracic spine is better in the head and foot direction but worse in the left and right direction compared with that of the lumbar spine. Instead of extending or narrowing the margin according to the BMI of patients with spinal metastasis, the margin must be changed according to the site of spinal bone metastasis.