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 To explore the risk factors and possible preventive measures of delayed postoperative intestinal paralysis(PPOI)in elderly(aged ≥75 years)patients with colorectal cancer.Methods This retrospective study included 333 patients with CRC who underwent laporascopic resection in General Department of Beijing Chaoyang Hospital from June 2016 to August 2023.There were 126 patients were enrolled in PPOI group and 207 patients were enrolled in non-PPOI group.The perioperative clinical characteristics of the patients were compared between PPOI group and non-PPOI group,and the risk factors of PPOI and potential preventive measures for them were investigated usingLogisticregression.Results The incidence of PPOI after laparoscopic surgery in elderly patients with colorectal cancer was 37.84％.The ages of the PPOI group and the non-PPOI group were(82.60±3.587)years and(80.38±3.847)years respectively.The rates of primary enterostomy during the operation were 20.63％and 9.66％,respectively,and the preoperative combined nutritional risks were 53.97％and 20.77％,respectively.The preoperative serum albumin levels were lower,which were(35.32±3.77)g/L and(38.36±3.91)g/L,respectively,and the preoperative hemoglobin levels were(104.47±20.31)g/L and(110.33±20.27)g/L,respectively.The intraoperative blood loss was(140.48±130.65)mland(98.26±56.45)ml,respectively.The patients who received enhanced recovery after surgery(ERAS)measures during the perioperative period were 14.29％and 75.85％,respectively.There was a statistically significant difference between the two groups(P＜0.05).The Logistic analysis showed that the risk factors for elderly patients with PPOI including increased age,preoperative nutritional risk,low preoperative albumin,increased intraoperative blood loss.Moreover,the implementation of ERAS protocols including preoperative nutritional support,multimodal low-opioid anesthesia,gastric tube removal and ground activity early after surgery may be the protective factors of PPOI.Conclusion The risk factors of PPOI for colorectal patients older than 75 years including increased age,preoperative nutritional risk,low preoperative albumin,increased intraoperative blood loss.The ERAS protocols including preoperative nutritional support,multimodal low-opioid anesthesia,gastric tube removal and ground activity early after surgery may be useful to prevent the occurrence of PPOI for elderly patients with colorectal cancer.

The dysregulation of cyclin-dependent kinase 12(CDK12),which may result from genomic alterations or modulation by upstream effectors,is implicated in cancer oncogenesis and progression.CDK12 over-expression or activation is sufficient to induce tumor initiation,recurrence,and therapeutic resistance.However,CDK12 may also exert tumor-suppressive functions in a context-dependent manner.Therefore,caution is warranted when targeting CDK12 in future clinical trials.A comprehensive elucidation of the dual roles and underlying mechanisms of CDK12 in carcinogenesis is urgently needed to advance pre-cision oncology.This review provides an overview of the current understanding of the dysregulation and biological roles of CDK12 in cancer.Subsequently,we systematically summarize the functions and mechanisms of the oncogenic and tumor-suppressive roles of CDK12 in different contexts.Finally,we discuss the potential of CDK12 as a novel therapeutic target and its implications in clinical oncology,offering insights into future directions for innovative cancer treatment strategies.

In order to comprehensively deepen the reform of medical insurance payment methods and explore the establishment of a medical insurance payment system with Chinese features,five representative cities have been selected for analysis:Jinhua City in Zhejiang Province,Xiamen City in Fujian Province,Suixi City in Anhui Province,TianjinCity,and Zhenjiang City in Jiangsu Province.These regions have been chosen due to their typical ambulatory care payment reform.It summarises the features of ambulatory care payment reforms in different regions regarding implementing agencies,covered crowds and types of illnesses,and payment methods.It is recommended that a reasonable scope of implementation be selected and the reform purpose and basic conditions should be taken into account to explore an appropriate ambulatory care payment reform.It is proposed that an innovative payment method reform be implemented to promote healthcare and prevention integration,and to incorporate a value-oriented paradigm,thereby realizing the benefits of the doctor and patient insurance.

Objective To explore the risk factors and possible preventive measures of delayed postoperative intestinal paralysis(PPOI)in elderly(aged ≥75 years)patients with colorectal cancer.Methods This retrospective study included 333 patients with CRC who underwent laporascopic resection in General Department of Beijing Chaoyang Hospital from June 2016 to August 2023.There were 126 patients were enrolled in PPOI group and 207 patients were enrolled in non-PPOI group.The perioperative clinical characteristics of the patients were compared between PPOI group and non-PPOI group,and the risk factors of PPOI and potential preventive measures for them were investigated usingLogisticregression.Results The incidence of PPOI after laparoscopic surgery in elderly patients with colorectal cancer was 37.84％.The ages of the PPOI group and the non-PPOI group were(82.60±3.587)years and(80.38±3.847)years respectively.The rates of primary enterostomy during the operation were 20.63％and 9.66％,respectively,and the preoperative combined nutritional risks were 53.97％and 20.77％,respectively.The preoperative serum albumin levels were lower,which were(35.32±3.77)g/L and(38.36±3.91)g/L,respectively,and the preoperative hemoglobin levels were(104.47±20.31)g/L and(110.33±20.27)g/L,respectively.The intraoperative blood loss was(140.48±130.65)mland(98.26±56.45)ml,respectively.The patients who received enhanced recovery after surgery(ERAS)measures during the perioperative period were 14.29％and 75.85％,respectively.There was a statistically significant difference between the two groups(P＜0.05).The Logistic analysis showed that the risk factors for elderly patients with PPOI including increased age,preoperative nutritional risk,low preoperative albumin,increased intraoperative blood loss.Moreover,the implementation of ERAS protocols including preoperative nutritional support,multimodal low-opioid anesthesia,gastric tube removal and ground activity early after surgery may be the protective factors of PPOI.Conclusion The risk factors of PPOI for colorectal patients older than 75 years including increased age,preoperative nutritional risk,low preoperative albumin,increased intraoperative blood loss.The ERAS protocols including preoperative nutritional support,multimodal low-opioid anesthesia,gastric tube removal and ground activity early after surgery may be useful to prevent the occurrence of PPOI for elderly patients with colorectal cancer.

Objective To explore the impact of preoperative fibrinogen levels on the 1-year adverse outcomes after endovascular revascularization in patients with diabetes complicated with lower extremity arteriosclerosis obliterans(LEASO).Methods We collected the baseline clinical data of 289 patients with diabetes complicated with LEASO,who were admitted to The First Affiliated Hospital of Xi'an Jiaotong University from May 2020 to December 2022 for endovascular revascularization.All patients were followed up for 13 to 24 months after interventional therapy,with the follow-up information including major adverse cardiovascular events(MACEs)such as all-cause death,acute myocardial infarction and acute stroke,as well as major adverse lower extremity events(MALEs)such as rest pain in the lower extremities,ulcers or skin defects,gangrene,reocclusion and amputation.A multivariable Cox regression model was used to analyze the related risk factors for adverse outcomes 1 year after endovascular revascularization in patients with diabetes complicated with LEASO,and receiver operating characteristic(ROC)curves were constructed to evaluate the predictive efficacy and optimal cutoff value of fibrinogen levels for endpoint events,and Kaplan-Meier survival curves were drawn.Sensitivity analysis was made to assess the differences in the impact of fibrinogen on endpoint events across various subgroups.Results We recruited a total of 289 patients(55 patients in MACEs and 234 in non-MACEs;68 patients in MALEs and 221 in non-MALEs),with a mean age of 67.6±9.3 years,including 215 males.Multivariate Cox regression analysis showed that elevated plasma fibrinogen was an independent risk factor for MACEs(HR=1.250,95％CI:1.053-1.484,P=0.011)and all-cause death(HR=1.297,95％CI:1.030-1.633,P=0.027)in the cohort followed up 1 year after interventional therapy,but had no significant impact on the occurrence of MALEs(P=0.625).Baseline plasma fibrinogen level 4.32 g/L was the optimal cutoff value for predicting MACEs(sensitivity=0.673,95％CI:0.582-0.767;specificity=0.688,95％CI:0.562-0.775)and all-cause death(sensitivity=0.679,95％CI:0.483-0.880;specificity=0.651,95％CI:0.465-0.755).The AUC for predicting MACEs and all-cause death after interventional therapy was 0.652(95％CI:0.564 2-0.739 1)and 0.619(95％CI:0.507-0.733),respectively.After a median follow-up of 14.03 months,patients with preoperative fibrinogen level ≥ 4.32 g/L had a significantly higher risk of MACEs and all-cause death compared to patients with preoperative fibrinogen＜4.32 g/L(P＜0.001),and there were no significant differences in different subgroups,including gender(male/female,interaction P=0.836),age(＜65 years/≥65 years,interaction P=0.211),smoking status(never smoked/current or former smoker,interaction P=0.779),chronic kidney disease(yes/no,interaction P=0.360),and heart failure(yes/no,interaction P=0.114).Conclusion Preoperative plasma fibrinogen≥4.32 g/L is an effective indicator for predicting MACEs and all-cause mortality following endovascular revascularization in patients with diabetes and LEASO.

Objective:To investigate the relationship between work fit, work stress, and occupational fatigue among medical staff, and to analyze the independent and interactive effects of work fit and work stress on occupational fatigue.Methods:A stratified random sampling method was employed to select 1162 employees from two top-tier hospitals as the research subjects. A questionnaire survey was conducted to collect information on basic demographics, work stress, work fit, and occupational fatigue. Chi-square test, analysis of variance, and logistic regression equations were used to evaluate the interactive effects of work fit and work stress on occupational fatigue.Results:This study included 1162 medical Homo sapiens personnel, among whom 884 were job-adapted and 362 were in a stressful state. Univariate logistic regression analysis showed that the risk of occupational fatigue significantly increased in the 40-49 age group ( OR=1.89) and the bachelor's degree group ( OR=2.52, P<0.01). Smoking and alcohol consumption were also associated with a significantly higher risk of occupational fatigue ( P<0.01). In terms of occupational characteristics, compared to clinical doctors, nurses and pharmaceutical/technical Homo sapiens personnel had lower risks ( OR=0.57, 0.43, P<0.05). Compared to high-ranking Homo sapiens personnel, those with no title or junior titles exhibited lower risks ( OR=0.51, 0.43, P<0.05). Working hours exceeding 55 hours per week and night shifts 3-4 times per week significantly increased fatigue risk ( OR=1.94, 1.90, P<0.05). Occupational stress and job discomfort were identified as risk factors for occupational fatigue ( OR=11.94, 3.26, P<0.05). Job adaptation and occupational stress exhibited a multiplicative interaction on physical fatigue after adjusting for confounders[ OR (95% CI) =0.43 (0.22, 0.84) ]. Additionally, job adaptation and occupational stress demonstrated an additive interaction on mental fatigue, with RERI (95% CI) =2.64 (0.11, 5.59), API (95% CI) =0.38 (0.08, 0.67), and SI (95% CI) =1.78 (1.01, 3.14). Medical Homo sapiens personnel in a stressful state had a 7.25-fold higher risk of fatigue compared to those not in a stressful state, while those with job discomfort had a 2.01-fold higher risk compared to those in an adapted state. Conclusion:Occupational stress and job fit have interactive effects on physical fatigue and mental fatigue in medical Homo sapiens personnel. A stressful state combined with job discomfort increases the risk of fatigue occurrence, providing a basis for developing intervention strategies for occupational fatigue among medical Homo sapiens personnel.

To explore the effect of Macleaya cordata extract(MCE)on growth performance and serum biochemistry of Sichuan White Geese,and to analyze its mechanism of action by network pharmacology and molecular docking technology combined with animal experiments verification.A total of 90 1-day-old healthy goslings were randomly divided into 5 groups with 18 goslings per group after 5 d of adaptive feeding.The control group(CON)was fed with basal diet,the antibiotic group(CTC)was supplemented with 450 mg/kg chlortetracycline premix,and the low MCE group(MCEL),medium MCE group(MCEM)and high MCE group(MCEH)were supple-mented with 200,300 and 400 mg/kg MCE,respectively.The experimental period was 21 d.On the basis of the above experiments,Network pharmacology and molecular docking technology were used to predict the core targets and signaling pathways of MCE to promote geese growth from the levels of antioxidant,immune and apoptosis,and the expression levels of the core target genes were detected by Real-time fluorescence quantitative PCR.The results showed that compared with the CON group,MCE supplementation could increase the average daily weight gain,decrease the ratio of feed to gain,significantly increase the contents of serum GH,T3,T4,TP and ALB(P＜0.05),and significantly decrease the contents of serum AST and TG(P＜0.05).Network pharmacology analysis predicted 2 active ingredient and 237 active ingredient targets,and concluded that the mechanism of MCE promoting the growth of Sichuan White Geese may be related to the role of 5 key targets such as SRC,HSP90AA1,CASP3,ESR1 and MAPK14,and the action of MAPK,apop-tosis and other signaling pathways.Molecular docking results showed that the active ingredients of sanguinarine and chelerythrine in MCE could act through MAPK14.The validation results of core targets showed that MCE could significantly reduce the mRNA expression levels of CytC,CASP2,CASP3 and CASP9 in spleen(P＜0.05)and significantly increase the mRNA expression levels of Mcl-1(P＜0.05).These results indicated that MCE could promote the growth performance of Si-chuan White Geese by regulating apoptosis,promoting the secretion of serum growth-related hor-mones and improving biochemical indicators.

Background Although current evidence suggests a link between outdoor air pollution and depressive symptoms, the effect of solid fuel use (a significant indoor air pollutant) on depressive symptoms in China's rural middle-aged and elderly population remains poorly understood. Objective To explore the association between solid fuel use for cooking and depressive symptoms among middle-aged and elderly people in rural areas of China, and to provide a basis for the prevention and control of depressive symptoms among residents in rural areas. Methods Data were obtained from the 2020 China Family Panel Studies (CFPS), depressive symptoms were assessed using 8-item Center for Epidemiologic Studies Depression Scale (CES-D), and cooking fuel type was self-reported. Subsequently, two-level binary unconditional logistic regression models were fitted to assess the impact of solid fuel use for cooking on depressive symptoms. Results A total of 7991 participants aged 45 years and older [mean age (58.52±9.18) years] were included in the study, with a mean CES-D score of 6.07±4.48 and a mean positive depressive symptoms rate of 34% (40.54% in the solid fuel group and 30.81% in the clean fuel group, χ²=74.40, P<0.001). After controlling for potential confounding covariates, rural middle-aged and elderly residents in the solid fuel group had a higher risk of reporting depressive symptoms than those in the clean fuel group (OR=1.36, 95%CI: 1.20-1.54), an association that was unaffected by participant characteristics and was positive across age and gender groups. Conclusion There is a positive association between solid fuel use for cooking and depressive symptoms in middle-aged and elderly people in rural areas of China. It is recommended that rural areas promote the upgrading of stoves through the installation of ventilation equipment or the switch from solid cooking fuel to clean fuel. This approach is directed toward reducing exposure to household air pollution, alleviating the prevalence of depression among middle-aged and elderly residents in vast rural areas, and enhancing their mental health.

OBJECTIVE To analyze and identify the metabolites of pirtobrutinib （PTN） in rats， and clarify the possible metabolic pathways of PTN in rats. METHODS Six rats were intragastrically administered with 10 mg/kg PTN suspension. Blood samples were collected from the rats 30 minutes before administration and at 0.25， 0.5， 1， 2， 4， 6， 8， 12， 24 hours after administration. Urine and feces samples were collected 12 hours before administration and 24 hours after administration. UHPLC- Orbitrap Exploris 240 system combined with Compound Discoverer 3.0 and Xcalibur 2.0 software were adopted for structural identification and metabolic pathway analysis of PTN metabolites in rat plasma， urine， and feces. RESULTS A total of 29 PTN metabolites were identified， including 17， 19 and 22 metabolites in plasma， urine and feces， respectively. The metabolic pathways of PTN mainly included oxidation， sulfation， glucuronidation， etc.， and its metabolites were mostly combination products of two or more different metabolic forms. In detail， a total of 26 metabolites were associated with phase Ⅰ metabolic reactions （14 oxidation metabolites， 9 reduction/dehydrogenation metabolites， 8 demethylation metabolites， and 5 hydrolysis metabolites）. Meanwhile， a total of 20 products were involved in phase Ⅱ metabolites （14 sulfation metabolites and 8 glucuronic acid binding metabolites）. CONCLUSIONS PTN exhibits a diverse range of metabolites in rat fecal samples， with the primary metabolic pathways being oxidation， sulfation， glucuronidation， and others.