Background/Aims: Biliary tract cancer (BTC) frequently harbors KRAS mutations, which are associated with resistance to traditional treatment and a poor prognosis. Synthetic lethality (SL) strategy may provide other targets of KRAS. Therefore, we aim to identify and validate potential therapeutic targets of KRAS for the treatment of BTC via SL. Methods: The dependency (DepMap) projects were used to predict the synthetic lethal gene of KRAS. FDA-approved anticancer drug library was applied to screen potential drugs effective against KRAS-mutant BTC. Furthermore, the synthetic lethal effects or corresponding mechanisms of potential genes and drugs on BTC were investigated using KRAS-mutant and KRAS-wild type BTC cell lines, patient-derived xenografts (PDX), and KRAS oncogene-driven tumor models, as well as other KRAS-mutant cancer cell lines. Results: Initially, we discovered that the loss of GATA2 reduced the viability of KRAS-mutant but not KRAS-wild-type BTC. Subsequently, the drug library screened out disulfiram, which primarily exerts a synthetic lethal effect by inhibiting interleukin-1β (IL-1β) in KRAS-mutant BTC. Mechanistically, GATA2 specifically enhanced the transcription of IL-1β to promote NF-κB signaling in KRAS-mutant BTC. IL-1β inhibition phenocopied GATA2 deficiency, leading to reduced KRAS-mutant BTC viability. These synthetically lethal effects were confirmed using PDX, a KRAS oncogene-driven tumor model, as well as in other KRAS-mutant cancer cell lines. Conclusions In summary, these results indicate that inhibiting GATA2/IL1β could be a therapeutic strategy in KRAS-mutant BTC and potentially other cancers.

Objectives:To investigate the mechanism underlying the protective effects of polydatin(PD)a-gainst oxygen-glucose deprivation(OGD)-induced neuronal injury and acute spinal cord injury(SCI).Methods:The mouse hippocampal neuronal cell line HT22 was used in in vitro experiments.The cytotoxicity of PD was assessed using the CCK-8 assay:cells were treated with 0,1,3,10,30 and 100μM PD for 24h to de-termine the safe concentration range.After OGD modeling(24h),cells were treated with different concentrations of PD(0,3,10,and 30μM),and cell viability was measured via CCK-8 to identify the optimal protective concentration.Autophagy markers(LC3 and P62)were detected by immunofluorescence and Western blot(WB);Autophagosome formation was observed using transmission electron microscopy;And the apoptosis rate was e valuated by TUNEL staining.For in vivo studies,an acute SCI model was established in C57BL/6 mice using Allen's impact method.Animals were divided into sham,SCI model,and PD treatment(20mg/kg)groups,with n=10 per group.Tissues were collected on 7d and 14d post-injury.Spinal cord pathology was examined by HE staining on 7d and 14d.Immunofluorescence was performed on 14d to evaluate glial scar formation(GFAP+),neuronal survival(MAP2+),cell proliferation(BrdU+),and autophagy level(LC3 Ⅱ).Results:In vitro results showed that 1-100μM PD did not exhibit significant cytotoxicity toward HT22 cells.The optimal concentration of PD was determined to be 10μ M after OGD induction.Compared with the OGD group,10μM PD significantly enhanced cell viability after OGD[OGD group:(54.63±3.90)％vs OGD+PD(1 0μ M)group:(84.35±1.38)％,P＜0.05]and effectively attenuated autophagy activation,as evidenced by a decreased LC3Ⅱ/Ⅰratio(OGD group:11.0±0.57 vs OGD+PD group:3.50±0.28,P＜0.05),increased P62 accumulation(OGD group:0.55±0.04 vs OGD+PD group:0.93±0.06,P＜0.05),reduced number of autolysosomes,and significantly lower apoptosis rate[OGD group:(35.33±2.6)％vs OGD+PD group:(19.67±1.76)％,P＜0.05].In vivo,HE staining confirmed that pathological damage in spinal cord tissue was markedly alleviated in the PD-treated group compared with the SCI model group.Immunofluorescence results indicated that PD treatment inhibited fibrous scar formation(SCI group:2.32±0.19mm2 vs PD group:1.07±0.24mm2,P＜0.05),reduced neuronal damage(SCI group:1.72±0.28mm vs PD group:0.93±0.12mm,P＜0.05)and promoted cell proliferation[SCI group:(16.14±2.24)％vs PD group:(39.09±3.16)％,P＜0.05],and downregulated LC3 Ⅱ expression(SCI group:62.81±5.25 vs PD group:34.09±3.98,P＜0.05).Conclusions:PD ameliorates neural damage by concurrently suppressing autophagy and apoptosis,providing a dual-pathway therapeutic strategy for SCI.

Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.

OBJECTIVE To investigate the positive test of high-risk human papillomavirus(HR-HPV)among the female population undergoing physical examination in Shenyang,analyze the influencing factors and establish and validate the risk prediction model.METHODS The data were collected from the female population who received HPV test in the physical examination center of a three-A hospital in the whole year of 2023.The prevalence rates of HR-HPV infections and subtypes were described,the influencing factors for the infections were identified.Uni-variate analysis and multivariate logistic regression analysis were performed for the influencing factors for positive test of HR-HPV,and the prediction model was established and validated.RESULTS Totally 6 130 out of 7 759 fe-male population who received HPV test were from Shenyang,the total positive rate of HR-HPV was 10.72％a-mong the population from Shenyang,11.11％among the population from other areas,and there was no signifi-cant difference.The population from Shenyang aged between 21 and 84 years old,with the mean age(48.58±11.64)years old.Among the local population who had the infections,80.21％were the single HPV infection,and 19.79％were multiple infections;HPV52 was the predominant subtype of HPV causing the infections,followed by HPV58 and HPV 16.The result of multivariate analysis showed age,smoking history,gynecological surgery history,allergic history,family annual income and sleep condition were the influencing factors for the positive HR-HPV.The prediction model was established based on the result of the multivariate analysis,the internal vali-dation of the model was carried out by modeling data and receiver operating characteristic(ROC)curves,the area under the curve(AUC)of the prediction model was 0.919,and 95％CI was 0.878 to 0.960,indicating that the prediction model had a high efficiency.CONCLUSIONS The positive rate of HR-HPV test is not relatively high among the physical examination female population in Shenyang,and the positive result is affected by a variety of factors.The population can be vaccinated for prevention and control based on the prediction model targeting to the non-variable factors such as age,meanwhile,the measures such as enhancement of health education,adjustment of health polies and interven-tion to health behaviors should be taken for other controllable factors.

Objective To explore the value of diffusion kurtosis imaging(DKI)in the diagnosis of Parkinson's disease with mild cognitive impairment(PD-MCI)patients.Methods A total of 18 patients with Parkinson's disease cognitive normal(PDN),22 patients with PD-MCI,and 24 healthy controls(HC)were prospectively included.All participants underwent DKI,and regions of interest(ROI)were selected in the substantia nigra,red nucleus,striatum,and posterior cingulate gyrus for post-processing.The diagnostic efficacy of DKI parameters on PD patients' cognitive status was analyzed by the receiver operating characteristic(ROC)curve.Results Compared with the PDN group,the PD-MCI group had a longer disease duration and a higher H-Y stage.Compared with the HC group,the PD-MCI group showed significantly lower mean kurtosis(MK),radial kurtosis(RK),axial kurtosis(AK),and fractional anisotropy(FA)values in the substantia nigra and posterior cingulate gyrus.In the PDN group,FA and MK values in the substantia nigra were significantly decreased,while FA values in the striatum and posterior cingulate gyrus were significantly increased(P＜0.05).Compared with the PDN group,the PD-MCI group showed significantly decreased DKI parameter values in the substantia nigra and posterior cingulate gyrus,and significantly decreased RK,AK,and FA values in the striatum(P＜0.05).The FA values of striatum,posterior cingulate gyrus and joint predictors were the most effective in the diagnosis of PD-MCI and the area under the curve(AUC)were 0.826,0.853 and 0.960,respectively.Conclusion DKI can detect microstructural changes in PD patients.Microstructural alterations in the striatum and posterior cingulate gyrus have an impact on early cognitive function changes in PD patients.FA demonstrate high sensitivity and specificity in the diagnosis of PD-MCI,and the combined diagnostic efficacy across multiple regions is even higher.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

Objective:To explore the chain mediating role of self-management efficacy and self-perceived burden between family function and decision fatigue in gastric cancer patients.Methods:Convenience sampling was used to select 219 gastric cancer inpatients in Xinxiang Central Hospital of Henan Province from June 2023 to December 2024 as study subjects. Family Assessment Device, Chinese version of the Strategies Used by People to Promote Health, Self-Perceived Burden Scale, and Decision Fatigue Scale were used to conduct the survey. The chain mediating effect were tested using Model 6 in the SPSS PROCESS program.Results:Family function, self-management efficacy, self-perceived burden and decision fatigue scores of gastric cancer patients were (73.08±8.47), (90.15±14.33), (31.64±6.55), and (19.23±3.05), respectively. Univariate analysis showed that age, education level, per capita monthly family income, tumor stage and treatment method were the influencing factors of decision fatigue in gastric cancer patients, and the difference was statistically significant ( P<0.05). Correlation analysis showed that family function was negatively correlated with self-management efficacy ( P<0.05) and positively correlated with self-perceived burden and decision fatigue ( P<0.05), and self-management efficacy was negatively correlated with self-perceived burden and decision fatigue ( P<0.05), and self-perceived burden was positively correlated with decision fatigue ( P<0.05), and the differences of the above were statistically significant. Mediating effect analysis showed that family function positively predicted decision fatigue in gastric cancer patients, with a direct effect of 0.117, accounting for 62.57% of the total effect. Self-management efficacy and self-perceived burden realized the mediating effect between family function and decision fatigue through three mediating pathways. Mediating pathway 1 was family function → self-management efficacy → decision fatigue, with an effect value of 0.032, which accounted for 17.11% of the total effect. Mediating pathway 2 was family function→ self-perceived burden → decision fatigue, with an effect value of 0.026, which accounted for 13.90% of the total effect. Mediating path 3 was family function → self-management efficacy → self-perceived burden → decision fatigue, with an effect value of 0.012, accounting for 6.42% of the total effect. The total mediating effect was 0.070, accounting for 37.43% of the total effect. Conclusions:Family function can not only directly influence decision fatigue in gastric cancer patients, but also indirectly influence decision fatigue through the mediating role of self-management efficacy and self-perceived burden. Decision fatigue of gastric cancer patients could be reduced in the future by improving family function, increasing patients' self-management efficacy, and reducing their self-perceived burden.

BACKGROUND:In the research and application of neural stem cells,cell culture and passage are key links,which directly affect the quality of cells and experimental results.It is of great significance to find the most suitable digestive enzymes that can maintain the biological characteristics of embryonic mouse spinal cord neural stem cells and enhance their passage efficiency.OBJECTIVE:To explore the most suitable digestive enzyme for passage of neural stem cells from the spinal cord of embryonic mice.METHODS:Microscopic dissection was used to isolate and extract spinal cord tissue from E14 d embryonic mice,which was cultured in DMEM/F12 serum-free medium containing epidermal growth factor,basic fibroblast growth factor,and B27.After spherulation,Nestin and Sox2 immunofluorescence identification was performed.During neural stem cell passage and culture,single-cell suspensions were prepared using trypsin,papain,and TrypLETM Express enzyme digestion combined with blow molding.The cell dispersion and spheroidization were observed,and passage 3 cells were stained with propidium iodide to detect cell death.Cell proliferation was detected by counting the total number of cells.Immunofluorescence staining,western blot assay and RT-PCR were used to detect the expression of Olig2,Tuj1,GFAP,and NeuN at the protein and mRNA levels and to identify cell differentiation.RESULTS AND CONCLUSION:After 72 hours of culture,E14 d embryonic mouse spinal cord tissue cells could form suspended neurospheres,which could be passaged after 5-7 days.Compared with trypsin and papain,TrypLETM Express enzyme combined with blow beating method was used for passage.The cell dispersion rate was high,the activity was good,and more NeuN-and Tuj1-positive neurons differentiated.This study optimized the culture and passaging process of neural stem cells,laying a foundation for further research on stem cell transplantation therapy for spinal cord diseases.

Objective:To explore the predictive value of preoperative CT angiography (CTA) for incomplete endothelialization of the occluder after left atrial appendage closure (LAAC) in patients with atrial fibrillation.Methods:The clinical data of 92 atrial fibrillation patients underwent LAAC in the Tangdu Hospital of Air Force Military Medical University from January 2022 to June 2024 were retrospectively analyzed. CTA examinations were performed both before operation and 3 months after operation. Before operation, the long diameter of left atrial appendage opening, short diameter of left atrial appendage opening, area of left atrial appendage opening, diameter of anchoring area and effective depth were measured. After operation, the condition of occluder endothelialization was evaluated, and the patients were divided into the completely endothelialization group and the incomplete endothelialization group. Multivariate Logistic regression analysis was used to analyze the independent risk factors of occluder incomplete endothelialization after LAAC in patients with atrial fibrillation. The predictive value of independent risk factors was evaluated by the receiver operating characteristic (ROC) curve.Results:Among the 92 patients, CTA 3 months after the operation showed that 58 cases had complete occluder endothelialization (complete endothelialization group), and 34 cases had occluder incomplete endothelialization (incomplete endothelialization group). Before operation, the long diameter of left atrial appendage opening, short diameter of left atrial appendage opening, area of left atrial appendage opening and diameter of anchoring area in incomplete endothelialization group were significantly larger than those in complete endothelialization group: (28.35 ± 1.77) mm vs. (26.21 ± 2.21) mm, (22.09 ± 2.01) mm vs. (20.86 ± 1.75) mm, (512.76 ± 63.35) mm 2 vs. (453.83 ± 75.39) mm 2 and (24.71 ± 2.50) mm vs. (23.12 ± 2.40) mm, and there were statistical differences ( P<0.01); there was no statistical difference in effective depth between the two groups ( P>0.05). Multivariate Logistic regression analysis result showed that the long diameter of left atrial appendage opening before operation was an independent risk factor for occluder incomplete endothelialization after LAAC in patients with atrial fibrillation ( OR = 2.141, 95% CI 1.217 to 3.768, P<0.01). ROC curve analysis result showed that the area under the curve of the long diameter of left atrial appendage opening before operation for predicting occluder incomplete endothelialization after LAAC in patients with atrial fibrillation was 0.768 (95% CI 0.674 to 0.862, P<0.01), the optimal cut-off value was 26.5 mm, the sensitivity was 88.2%, and the specificity was 55.2%. Conclusions:A larger long diameter of left atrial appendage opening before operation can lead to occluder incomplete endothelialization after LAAC in patients with atrial fibrillation.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.