Background: Alzheimer’s disease (AD) is the most prevalent neurodegenerative disorder. Cellular drug screening represents a key approach in the preclinical discovery of AD therapeutics. Moreover, selecting an appropriate cellular model is essential for exploring the mechanisms underlying AD pathogenesis. Objective: This study aimed to establish a progressive strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery for the development of novel therapeutics targeting AD. Methods: A novel AD12 cell line stably expressing the double-transgenic genes amyloid precursor protein (APP)695/presenilin 1(PS1)-ΔE9 was first constructed and subjected to lipopolysaccharide (LPS) stimulation. The resulting LPS-induced neuroinflammatory response was characterized using metabolomic analysis. Subsequently, 16 alkaloids derived from traditional Chinese medicine(TCM) were screened with the LPS-AD12 cell model. Molecular docking was then performed to assess the binding affinity of benzoylaconitine (BAC) with APP and PS1 proteins. A targeted metabolomics approach focusing on amino acids and the tricarboxylic acid (TCA) cycle was applied to BAC-treated LPS-AD12 cells. Finally, aconite extract was administered to five-familial-AD-mutations(5xFAD) transgenic mice, and therapeutic efficacy was evaluated through phenotypic and metabolomic analyses. Results: LPS stimulation induced neuroinflammation in AD12 cells, characterized by elevated levels of interleukin-6, amyloid-β (Aβ ), and PS1. The primary metabolic pathways affected in AD12 cells included alanine, aspartate, and glutamate metabolism, as well as arginine biosynthesis. Notably, BAC significantly reduced the levels of Aβ and interleukin-6 in LPS-AD12 cells. Furthermore, molecular docking analyses revealed a strong binding affinity of BAC with APP and PS1 proteins. Targeted metabolomics analysis indicated that the pharmacological activity of BAC was primarily associated with its anti-inflammatory effects, modulation of the cholinergic signaling pathway, and regulation of energy metabolism. Behavioral experiments in 5xFAD mice, together with immunofluorescence assessments of brain tissues (amyloid-β, glial fibrillary acidic protein, and ionized calcium-binding adapter molecule 1), demonstrated that aconite extract treatment significantly alleviated AD-related symptoms. In addition, metabolomics-based mechanistic analyses revealed that serine, acetylcholine, ornithine, and gamma-aminobutyric acid exhibited consistent trends in both brain tissue and serum samples from treated mice. Conclusion: This study established a novel AD12 cell model and an effective, reliable LPS-induced cell-based screening system. Additionally, we proposed a new drug discovery strategy integrating cell model construction, monomer evaluation, and effective component group-based drug discovery. This screening system has the potential to accelerate the identification of anti-AD agents and provides a feasible framework for subsequent in-depth investigations into new drug development from TCM, thereby contributing to the modernization of TCM-derived therapeutics.

Objective:To assess the levels of circRNA and N6-methyladenosine(m6A)methylation in hippocampal tissue of Alzheimer's disease(AD)mice and wild-type(WT)mice, and to explore the potential role of circRNA m6A methylation modification in the pathogenesis of Alzheimer's disease.Methods:Differentially expressed circular RNAs(circRNAs)in the hippocampal tissue of APPswe/PS1dE9(APP/PS1)double-transgenic Alzheimer's disease(AD)model mice and matched wild type mice were identified using RNA high-throughput sequencing(RNA-seq).Subsequently, Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted on the source genes of these circRNAs.Validation of the RNA-seq results was performed using real-time quantitative PCR(qRT-PCR).Furthermore, the SRAMP website was utilized to predict potential m6A modification sites on the differentially expressed circRNAs.Finally, circRNAs with high confidence were validated and analyzed through methylated RNA immunoprecipitation-quantitative PCR(MeRIP-qPCR).Results:In comparison to the control group, 21 circRNAs exhibited differential expression in the APP/PS1 group, with 15 being up-regulated and 6 down-regulated.GO analysis revealed that the source genes of these differentially expressed circRNAs were primarily associated with synapse structure and function, while KEGG analysis indicated their involvement in the Hippo signaling pathway and Lipid metabolism.qRT-PCR results confirmed that the expression patterns of the differentially expressed circRNAs were consistent with the RNA-seq findings.Specifically, the expression of mmu-Bai3_0007 in the APP/PS1 group was significantly lower than that in the control group(0.033±0.002 vs.1.006±0.136, t=12.38, P<0.01), and the m6A methylation level of mmu-Bai3_0007 was also notably reduced in the APP/PS1 group compared to the control group(0.144±0.018 vs.0.431±0.087, t=5.62, P<0.01). Conclusions:The m6A methylation modification of Bai3circRNA could play a role in Alzheimer's disease by regulating the expression of Bai3 circRNA.

Objective:To assess the levels of circRNA and N6-methyladenosine(m6A)methylation in hippocampal tissue of Alzheimer's disease(AD)mice and wild-type(WT)mice, and to explore the potential role of circRNA m6A methylation modification in the pathogenesis of Alzheimer's disease.Methods:Differentially expressed circular RNAs(circRNAs)in the hippocampal tissue of APPswe/PS1dE9(APP/PS1)double-transgenic Alzheimer's disease(AD)model mice and matched wild type mice were identified using RNA high-throughput sequencing(RNA-seq).Subsequently, Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted on the source genes of these circRNAs.Validation of the RNA-seq results was performed using real-time quantitative PCR(qRT-PCR).Furthermore, the SRAMP website was utilized to predict potential m6A modification sites on the differentially expressed circRNAs.Finally, circRNAs with high confidence were validated and analyzed through methylated RNA immunoprecipitation-quantitative PCR(MeRIP-qPCR).Results:In comparison to the control group, 21 circRNAs exhibited differential expression in the APP/PS1 group, with 15 being up-regulated and 6 down-regulated.GO analysis revealed that the source genes of these differentially expressed circRNAs were primarily associated with synapse structure and function, while KEGG analysis indicated their involvement in the Hippo signaling pathway and Lipid metabolism.qRT-PCR results confirmed that the expression patterns of the differentially expressed circRNAs were consistent with the RNA-seq findings.Specifically, the expression of mmu-Bai3_0007 in the APP/PS1 group was significantly lower than that in the control group(0.033±0.002 vs.1.006±0.136, t=12.38, P<0.01), and the m6A methylation level of mmu-Bai3_0007 was also notably reduced in the APP/PS1 group compared to the control group(0.144±0.018 vs.0.431±0.087, t=5.62, P<0.01). Conclusions:The m6A methylation modification of Bai3circRNA could play a role in Alzheimer's disease by regulating the expression of Bai3 circRNA.

Objective:To observe the safety of intravitreal injection of triamcinolone acetonide (TA) combined with partial anterior pars plana vitrectomy (PPV) for cataract with severe vitreous hemorrhage.Methods:A retrospective case-control study. A total of 60 patients (60 eyes) with cataract and severe vitreous hemorrhage were included from June 2017 to June 2019 in Xi’an People’s Hospital (Xi’an Fourth Hospital). There were 32 males (32 eyes) and 28 females (28 eyes), with a mean age of 64.8 years. The eyes were randomly divided into intravitreal injection TA group (TA group) and non-TA injection group (control group), with 30 eyes in each group. Before phacoemulsification, 0.05-0.10 ml TA was injected into the vitreous cavity in the TA group and the vitreous besides the vitrectomy channel was removed. The eyes of the control group underwent conventional cataract phacoemulsification combined with PPV. The follow-up time after surgery was ≥6 months. The rate of success continuous circular capsulorhexis, posterior capsule rupture, and intraocular lens (IOL) implantation in the capsular bag were compared between the two groups. The statistical data were compared with χ2 test. Results:For all eyes in the TA group, after intravitreal injection of TA combined with partial anterior PPV, the white reflection of the fundus can clearly show the anterior and posterior capsule of the lens. Continuous circular capsulorhexis was completed, the posterior capsule was not broken, and the IOL was successfully implanted inside the capsular bag. Among the 30 eyes of the control group, 4 eyes did not complete continuous circular capsulorhexis. Radial tear occurred during capsulorhexis, and capsulorhexis was used to complete capsulorhexis. In 5 eyes with posterior capsule rupture, the posterior capsule occurred in 3 eyes during phacoemulsification, and injecting the lens cortex caused posterior capsule repture in 2 eyes; the IOL was implanted in the ciliary sulcus and the capsular bag in 4 and 1 eyes, respectively. Whether the consecutive capsulorhexis was successful ( χ2=4.286), whether the posterior capsule was ruptured ( χ2=5.455), whether the IOL was implanted in the capsular bag ( χ2=4.286), the differences in the number of eyes between the two groups were statistically significant ( P= 0.038, 0.020, 0.038). At the last follow-up, no special complications occurred. Conclusions:Intravitreal injection of TA combined with partial anterior PPV can improve the visibility of the anterior and posterior lens capsule and lens nucleus during phacoemulsification in patients with cataract and severe vitreous hemorrhage. The surgical success rate is high, the complications are few, and the safety is good.

Objective:To investigate the colonscopy screening interval among patients with negative colonscopy.Methods:We selected 14 606 participants who completed the baseline and 3-year or 5-year colonoscopy examinations in the American Prostate, Lung, Colorectal, and Ovarian (PLCO) dataset as the target population. Sociodemographic characteristics (i.e., sex, age, marital status, race, and smoking), lifestyle, family history of cancer, and family history of colorectal cancer were collected. Cochran-Armitage trend analysis was used to examine whether the rate of positive cases (colorectal cancer, advanced adenoma, adenoma, and hyperplastic polyp) was increased with the length of screening interval. We compared the differences in number of detected cases, positive rates, and proportions of 3-year and 5-year screening interval strategies using internal standardization method.Results:The age of the population was （61.9±5.2） years and over half of them were males (54.4%) and 46.2% had family cancer history. The mean screening interval between the first and second endoscopies was （1 639.1±320.9） days. A total of 1 716 cases had positive endoscopic findings. With the screening interval extended, rate of the screened positive cases was also increased ( P for trend<0.001). After standardized by the internal standardized population (14 606), 17.99 and 11.57 colorectal cancer cases and 177.37 and 240.35 advanced adenoma cases were detected by 3-year and 5-year screening interval strategies, respectively. Conclusion:Based on the initial screening negative population of colonoscopy in the United States, the 3-year screening interval strategy could detect a relatively large number of colorectal cancer cases, but its health and economic evaluation needs to be further explored.

Objective:To investigate the colonscopy screening interval among patients with negative colonscopy.Methods:We selected 14 606 participants who completed the baseline and 3-year or 5-year colonoscopy examinations in the American Prostate, Lung, Colorectal, and Ovarian (PLCO) dataset as the target population. Sociodemographic characteristics (i.e., sex, age, marital status, race, and smoking), lifestyle, family history of cancer, and family history of colorectal cancer were collected. Cochran-Armitage trend analysis was used to examine whether the rate of positive cases (colorectal cancer, advanced adenoma, adenoma, and hyperplastic polyp) was increased with the length of screening interval. We compared the differences in number of detected cases, positive rates, and proportions of 3-year and 5-year screening interval strategies using internal standardization method.Results:The age of the population was （61.9±5.2） years and over half of them were males (54.4%) and 46.2% had family cancer history. The mean screening interval between the first and second endoscopies was （1 639.1±320.9） days. A total of 1 716 cases had positive endoscopic findings. With the screening interval extended, rate of the screened positive cases was also increased ( P for trend<0.001). After standardized by the internal standardized population (14 606), 17.99 and 11.57 colorectal cancer cases and 177.37 and 240.35 advanced adenoma cases were detected by 3-year and 5-year screening interval strategies, respectively. Conclusion:Based on the initial screening negative population of colonoscopy in the United States, the 3-year screening interval strategy could detect a relatively large number of colorectal cancer cases, but its health and economic evaluation needs to be further explored.

Objective To investigate the status of disease perception and empirical avoidance in patients with breast cancer surgery, and to analyze the correlation between them. Methods A total of 100 cases of breast cancer patients in the breast surgery department of Qingdao Municipal Hospital from June 2015 to October 2016 were selected by convenience sampling method and investigated with Perceived Benefits of Diagnosis and Treatment of Breast Cancer (PB-DT-BC) and Acceptance and Action Questionnaire-2nd Edition (AAQ-Ⅱ). Pearson correlation analysis was adopted to analyze the correlation between the disease perception and empirical avoidance. Results The total score of PB-DT-BC of the patients with breast cancer was (2.40±0.36), which was at a moderate level. The total score of empirical avoidance was (5.57±0.60), which was at a moderate to upper level. It showed a negative correlation between the disease perception and empirical avoidance (r=-0.675,P<0.05). Conclusions The perceived benefits of disease in patients with breast cancer surgery is low, which is closely related to the empirical avoidance. Improving the perceived benefits of disease can decrease the empirical avoidance in patients.

Objective To understand the nursing working environment and working-family conflict in the operating room nurses,and analyze the correlation between the two.Method Tolly 190 nurses in the operating room were surveyed by general information questionnaire,PES-NWI (the practice environment scale of the nursing work index) and Work-Family Conflict Scale.Results The total nursing work environment (95.48 ± 6.34) averaged (3.08 ± 0.44),and the total score of working-family conflict was (56.67 ± 4.56),averaged (3.15 ±0.48).There was a negative correlation between nurses' perceived nursing work environment and work-family conflict(P＜0.05).Conclusions Nurses in the operating room think that the operating environment of the operating room is at the middle-high level;the nurses'working and family conflict is at the middle-high level,and the nurses' perceived nursing work environment is the main factor that affects the work-family conflict of the nurses in the operating room.Nursing managers should pay attention to the creation of good nursing work environment,improve the operating room nurses recognition of the work environment,so as to effectively alleviate the conflict of nurses work and family.

OBJECTIVE:To establish a method for the plasma concentration determination of carboplatin,and study the phar-macokinetics of carboplatin in female rats after intravenous injection and intraperitoneal injection. METHODS:HPLC was per-formed on the column of Agilent TC-C18 with mobile phase of methanol-water(5:95,V/V)at a flow rate of 1.0 mL/min,detection wavelength was 229 nm,and column temperature was 25 ℃. The inner standard was 5-bromouracil,and injection volume was 20 μL. 24 SD rats were randomly divided into 4 groups,6 in each group. The rats were intravenously injected and intraperitoneally in-jected carboplatin 20,40 mg/kg respectively. 0.5 mL blood sample was taken from eyes before administration and after administra-tion of 0.25,0.5,1,1.5,2,4,6,8,10,12 h. The plasma concentration of carboplatin was determined,and DAS 2.0 was used to calculate the pharmacokinetic parameters. RESULTS:The linear range of carboplatin in plasma was 0.30-60.00 μg/mL (r=0.9991);RSDs of intra-day,inter-day precision were lower than 10%(n=5);RSD of peak area in stability test was lower than 10%(n=5);method recovery was 98.7%-102.4%(RSD≤6.08%,n=5),and extraction recovery was 83.38%-85.45%(RSD≤5.97%,n=5). AUC0-12 h of carboplatin 20,40 mg/kg by intravenous injection and intraperitoneal injection in female rats were (15.503 ± 4.172),(23.402 ± 4.266),(6.716 ± 2.306),(9.384 ± 2.205)μg·h/mL;AUC0-∞ were (16.424 ± 4.846),(23.404 ± 4.266),(6.790±2.378),(9.765±2.095)μg·h/mL;t1/2z were(1.246±0.765),(0.394±0.058),(0.513±0.156),(0.884±0.460) h;and tmax were(0.700±0.274),(0.400±0.335),(0.542±0.368),(0.833±0.289)h,respectively. CONCLUSIONS:The meth-od is simple,economic and accurate,with suitable internal standard,and can be used for the plasma concentration determination of carboplatin in female rats and the pharmacokinetic studies.

Objective:To observe the effect of Er Chen Tang on CYP2E1 and mitochondrial energy metabolism in nonalcoholic fat-ty liver disease ( NAFLD) to explore the role of Pinellinae Rhizoma Praeparata ( PRP) and Citri reticulatae pericarpium ( CRP) in the treatment of nonalcoholic fatty liver disease. Methods:Er ChenTang and the prescription without PRP or CRP was respectively given the animal models by gastric gavage. The serum levels of ALT, AST, triglyceride, cholesterol, SOD and MDA in hepatic tissue, and the contents of liver tissue CYP2E1 and ATP were detected in the mice. Results:The CYP2E1 levels in NAFLD mice increased signif-icantly with abnormal mitochondrial energy metabolism. Compared with those in the model group, the levels of ALT, AST, triglyceride and cholesterol were significantly reduced by Er Chen Tang, meanwhile, the content of CYP2E1 was reduced and also restored liver en-ergy metabolism. The treatment effect significantly decreased when the lack of PRP or CRP, and the ability of restoring liver mitochon-drial energy metabolism of Er Chen Tang decreased significantly when the lack of PRP (P<0. 05). After the removal of CRP, the in-hibition ability of Er Chen Tang to CYP2E1 levels significantly decreased (P<0. 05). Conclusion:Er Chen Tang can effectively im-prove nonalcoholic fatty liver diseases, and effectively reduce the content of CYP2E1 in liver tissue of mice and restore the mitochondri-al energy metabolism.