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.

Objective This study aims to investigate and analyze the distribution of MN blood type among ethnic minorities in China. Methods Through a systematic retrieval of the 981 literature related to MN blood group distribution, 120 literature, meeting the criteria of this study, with complete data were selected. The literature covers 49 ethnic minorities. SPSS 26 statistical software was used to analyze the data. Results The results showed that among the 49 ethnic minorities in China, the phenotype distribution of MN blood type was MN>MM>NN, with proportions of 42.54%, 41.86%, and 15.06% respectively. The gene frequency for MN blood type exhibited a trend of m>n, with a gene frequency of m being 0.6313 and n being 0.3687. Cluster analysis divided the Chinese ethnic minorities into three groups based on the gene frequency for m, showing the characteristics of Group I>Group II>Group III. Conclusion The MN blood type characteristics in Chinese ethnic minorities show a higher frequency of the M gene compared to the N gene. The frequency of the M gene is higher in southern ethnic minorities than in northern ones. There are significant differences between southwestern ethnic minorities and the Han nationality, but no differences with long-term mixed/settled Han populations.
Humans ; China/ethnology* ; Minority Groups ; Ethnicity/genetics* ; Gene Frequency ; Asian People/genetics* ; Blood Group Antigens/genetics*

Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

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.

Objective To explore the correlation between remnant cholesterol level and CI-AKI in elderly patients with ACS and concomitant T2DM following PCI.Methods A retrospective case-control study was conducted on 759 elderly T2DM patients with ACS undergoing PCI in the Affi-liated Hospital of Xuzhou Medical University between January 2020 and December 2023.The pa-tients were randomly allocated into a training group(n=608)and a validation group(n=151)in a ratio of 8∶2.Based on CI-AKI diagnostic criteria,the training group was further stratified into a CI-AKI subgroup(n=95)and a non-CI-AKI subgroup(n=513).Multivariate logistic regression analysis was used to identify the independent risk factors for CI-AKI.Results The remnant cho-lesterol level was significantly higher in the CI-AKI subgroup than the non-CI-AKI subgroup[0.78(0.51,1.07)mmol/L vs 0.57(0.40,0.81)mmol/L,P＜0.01].Red blood cell count,remnant cholesterol,age,uric acid,log-transformed systemic immune-inflammatory(SII)index,and cysta-tin C were identified as independent predictors for CI-AKI in T2DM patients with ACS after PCI(P＜0.01).ROC curve analysis demonstrated that the model combining remnant cholesterol with other independent predictors exhibited superior discriminative performance in both the training group(AUC=0.818,95％CI:0.773-0.863)and validation group(AUC=0.787,95％CI:0.686-0.889)when compared to the models excluding remnant cholesterol(P＜0.01).Conclusion Rem-nant cholesterol is an independent predictor of CI-AKI in elderly patients with ACS and T2DM af-ter PCI.A risk prediction model based on remnant cholesterol and other independent risk factors demonstrates enhanced predictive performance for postoperative CI-AKI in the patients.

Objective:To assess the effectiveness of Buzzy? device in alleviating pain and anxiety in children during puncture-related operation,and to provide more comprehensive intervention option for puncture-related management of children.Methods:Randomized controlled trials(RCTs)about the application of Buzzy? device in puncture-related pain and anxiety of children were searched from China National Knowledge Infrastructure(CNKI),Wanfang,VIP,PubMed,Embase and Web of Science(WOS)databases.The retrieval time was from the establishment of the database to July 2023.In the interventions measures,the control groups used different methods that included standard care group,ShotBlocker device group and distraction card group.The obtained 528 relevant literatures were implemented literature selection and data extraction by two independent reviewers according to the established inclusion and exclusion criteria.RevMan5.4 and Stata17.0 software were adopted to conduct Meta-analysis for the pain and anxiety of main outcome indicators,and the success rate of the first puncture,parents'satisfaction and adverse events of secondary outcome indicators in the literatures.Results:A total of 20 eligible studies that could meet the conditions were included,involving 2 876 children,including 1 110 children in the Buzzy? device group and 1 766 children in the control group.The Buzzy? device group was better than the standard care group and the distraction card group in alleviating puncture-related operation pain in children,and the differences of that among them were statistical significance(Z=7.18,2.13,P<0.05),and was better than the standard care group in anxiety,and the difference was statistical significance(Z=5.00,P<0.05).The Buzzy? device group and the distraction card group were better than standard care group and Shotblocker device group in satisfaction,and the difference was significant(x2=63.236,P<0.05).The interventions in the Buzzy? device group and other groups did not have effect on the success rate of the first puncture and incidence of adverse event.Conclusion:The Buzzy? device can alleviate pain and anxiety of children during puncture operation,and improve children's satisfaction for puncture operation.It does not affect the success rate of puncture,and there is not adverse events in its operation.

Objective:To investigate the relationship between sarcopenic obesity(SO), defined by various obesity indicators, and mortality risk in older adults based on a prospective cohort from multiple nursing homes.Methods:Sarcopenia was diagnosed according to the 2019 consensus of the Asian Working Group for Sarcopenia.Obesity was defined using six different indicators: waist circumference(WC), waist-hip ratio(WHR), waist-height ratio(WHTR), body mass index(BMI), visceral fat area(VFA), and percentage of body fat(PBF).A prospective cohort of adults aged 60 and above was established across 15 nursing homes in Zigong City, with annual follow-ups on survival status conducted over two consecutive years.Cox proportional hazards regression models were employed to analyze the association between SO, defined by different obesity indicators, and mortality risk, calculating hazard ratios( HR)and their 95% confidence intervals( CI). Results:A total of 695 older adults were included in the study, of whom 67.1% were male.During the 2-year follow-up period, 88 participants died.The prevalence of SO as defined by WC, WHR, WHTR, BMI, VFA, PBF was found to be 17.6%, 30.1%, 48.9%, 3.0%, 28.3%, and 58.7%, respectively.When SO was defined using WC, WHR, WHTR, BMI, and VFA, there was no statistically significant difference in mortality rates between the SO and non-SO groups.However, when defined by PBF, the SO group exhibited a significantly higher mortality rate compared to the non-SO group(16.9% vs.6.6%, P<0.01).Cox proportional hazards regression analysis revealed that, compared to the non-SO group, the SO group defined by PBF had a significantly increased mortality risk( HR=2.81, 95% CI: 1.67-4.73, P<0.001).After adjusting for potential confounding factors, the mortality risk for the SO group remained significantly higher than that of the non-SO group( HR=1.97, 95% CI: 1.14-3.38, P=0.015). Conclusions:The prevalence of SO varies significantly across different obesity indicators.SO defined by PBF is significantly associated with mortality risk in nursing home residents.This study provides new evidence for further optimizing the diagnostic criteria for SO in this population.

Objective:To analyze the infection status of HBV among the healthy population in Tongchuan city in 2023, and compare it against the data from 2006, 2014, and 2020, thereby elucidating epidemiological trends and providing reference for optimizing hepatitis B prevention and control strategies in Tongchuan city.Methods:By using the method of multi-stage stratified random sampling, healthy people aged 1-59 from eight villages/communities in the four districts and counties of Tongchuan city were selected for on-site investigation and blood sample collection. ELISA was used to detect the five hepatitis B markers of the survey respondents, including HBsAg and HBsAb, and the seroprevalence rates of the populations with different characteristics were analyzed. Statistical analysis was performed using Chi-square test.Results:In 2023, a total of 1 926 healthy people were surveyed. After standardization, the positive rates of HBsAg, HBsAb, and HBV, and the rate of all five hepatitis B markers being negative were 0.56%, 54.57%, 26.77%, and 33.05%, respectively. Children aged 1-4 had the lowest positive rates of HBsAg, HBcAb, HBeAb, and HBV, as well as the lowest rate of all five hepatitis B markers being negative, while they had the highest positive rate of HBsAb. In the population aged 1-29, the positive rates of HBsAg and HBV, and the rate of all five hepatitis B markers being negative showed an upward trend with increasing age, while the positive rate of HBsAb showed a downward trend. In particular, the positive rate of HBsAb dropped sharply in the 5-14 age group. In 2023, the positive rate of HBV in the population aged 1-29 was 10.93% (111/1 016), and the rate of all five hepatitis B markers being negative was 38.39% (390/1 016), both lower than the results of the previous three surveys. The positive rate of HBsAb was 56.69% (576/1 016) in people aged 1-29 in 2023, which was higher than the results of the previous three surveys.Conclusions:Although certain achievements have been made in the prevention and control of hepatitis B in Tongchuan city in 2023, with more than one-third of the population showing all negative results in the five hepatitis B tests, the positive rate of HBsAb in the 5-14 age group has dropped sharply. Further actions required for hepatitis B prevention and control in Tongchuan city include optimizing the childhood immunization schedule for national immunization program vaccines, implementing booster programs for adolescents, and developing tailored immunization strategies for adults.

Objective:To explore the potential role of lipoprotein-associated phospholipase A2 (Lp-PLA2) and components of metabolic syndrome (MS) in the early progression of carotid arteriosclerosis.Methods:The study was a cross-sectional study. Urban participants undergoing routine health check-ups were enrolled from all 11 prefecture-level cities in Zhejiang Province between January and December 2022. General clinical information was obtained through interviews, and data on MS was collected from the clinical health examinations. Serum Lp-PLA? levels were measured in all participants. All participants were divided into 3 groups according to the results of the carotid ultrasound: the normal group, the intima thickening group with carotid intima thickening change and the plaque group. Multivariable logistic regression models were used to evaluate the associations of Lp-PLA2, MS, and the components of MS with early carotid atherosclerosis. Receiver operating characteristic (ROC) curve analyses were performed to assess the diagnostic performance of combining Lp-PLA2 with MS and the cumulative number of MS components for early carotid atherosclerosis.Results:A total of 4 009 urban adults undergoing routine health check-ups were enrolled (mean age (48.9±8.46) years, 2 665(66.5 %)male). Of these, 1 398 were in the normal group, 1 650 in the intima thickening group, and 961 in the plaque group. Multivariable logistic regression demonstrated that Lp-PLA2 was independently associated with early carotid atherosclerosis ( OR=1.34, 95% CI: 1.11-1.63, P=0.003). Lp-PLA2 also showed independent positive associations with both carotid intima thickening and carotid plaque formation, with the latter being more pronounced (both P<0.05). MS was independently and positively associated with early carotid atherosclerosis ( OR=1.48, 95 % CI: 1.20-1.84, P<0.001), as well as with intima thickening and carotid plaque formation, with the association being stronger for the latter (both P<0.05). Furthermore, the strength of the association increased progressively with the number of MS components ( P<0.001), especially for carotid plaques formation (both P<0.001). Multivariable logistic regression revealed that, compared with individuals without MS and low Lp-PLA2 levels, the risk of early carotid atherosclerosis was increased in those with high Lp-PLA2 alone, MS alone, or both conditions concurrently, with the highest risk observed when both were present (all P<0.05). ROC analyses demonstrated that the combination of elevated Lp-PLA2 with 3, 4, or 5 MS components yielded good diagnostic performance for early carotid atherosclerosis ( AUC=0.869, 0.888, and 0.889, respectively), intima thickening ( AUC=0.844, 0.860, and 0.845, respectively), and carotid plaque formation ( AUC=0.899, 0.924, and 0.968, respectively) in urban health-screening participants. Conclusions:Lp-PLA2, MS, and the number of MS components were independently and positively associated with early carotid atherosclerosis in urban health chek-up populations. The combination of MS components and Lp-PLA2 provided favorable diagnostic performance for the detection of early carotid atherosclerosis.

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.