The development of atherosclerosis （AS） is closely associated with endothelial dysfunction and mitochondrial metabolic abnormalities. These alterations are fundamentally characterized by metabolic reprogramming， in which the tricarboxylic acid （TCA） cycle serves as a central regulatory hub. Disruption of the homeostasis of TCA cycle intermediates， including succinate， α -ketoglutarate， and itaconate， contributes to AS progression through multiple mechanisms， such as the modulation of oxidative stress， inflammation， and glucose and lipid metabolic networks. Owing to their multi-target and multi-pathway regulatory properties， bioactive compounds derived from traditional Chinese medicine （TCM） have demonstrated unique advantages in correcting metabolic dysregulation. Flavonoids， glycosides， and other bioactive constituents of TCM can target key enzymes and metabolites involved in the TCA cycle， thereby restoring aberrant metabolic reprogramming， alleviating endothelial injury induced by oxidative stress， inflammation， and glucose and lipid metabolic disorders， ultimately improving metabolic homeostasis and attenuating the progression of AS.

The development of atherosclerosis （AS） is closely associated with endothelial dysfunction and mitochondrial metabolic abnormalities. These alterations are fundamentally characterized by metabolic reprogramming， in which the tricarboxylic acid （TCA） cycle serves as a central regulatory hub. Disruption of the homeostasis of TCA cycle intermediates， including succinate， α -ketoglutarate， and itaconate， contributes to AS progression through multiple mechanisms， such as the modulation of oxidative stress， inflammation， and glucose and lipid metabolic networks. Owing to their multi-target and multi-pathway regulatory properties， bioactive compounds derived from traditional Chinese medicine （TCM） have demonstrated unique advantages in correcting metabolic dysregulation. Flavonoids， glycosides， and other bioactive constituents of TCM can target key enzymes and metabolites involved in the TCA cycle， thereby restoring aberrant metabolic reprogramming， alleviating endothelial injury induced by oxidative stress， inflammation， and glucose and lipid metabolic disorders， ultimately improving metabolic homeostasis and attenuating the progression of AS.

Objective:To construct the gemcitabine resistant cell lines of human pancreatic cancer cell line (PANC1) and mouse pancreatic cancer cell line (PANC02), and to investigate their biological behavior changes.Methods:Gemcitabine-resistant cell lines PANC1-GR of human pancreatic cancer and PANC02-GR of mouse pancreatic cancer were induced by concentration gradient increment method. Cell count assay (CCK-8), flow cytometry, cell scratch assay and Transwell assay were used to detect the drug resistance, proliferation, cell cycle, migration and invasion of the four groups of cell lines. The drug-resistant cells were also compared with the parent cells.Results:The resistance indices of PANC1-GR and PANC02-GR were 153.3 and 185.4, respectively. The results of CCK-8 showed that with the increase of gemcitabine concentration, the proliferation of resistant cells changed significantly compared with parental cells, the population doubling time of PANC1-GR was significantly shorter than that of PANC1 (1.5±0.1) d vs (2.4±0.2) d ( t=8.00, P<0.001). The proportion of cells in S and G2/M phase increased, and the proportion of cells in G0/G1 phase decreased. The cell scratch and Transwell experiments indicated that the 24h mobility of PANC1-GR and PANC02-GR was higher than that of parent cells (47.6±2.4)% vs (28.7±6.3)% and (53.6±3.2)% vs (30.1±1.4)%, the number of individual field (200 times magnification) penetrating membrane cells was also higher than that of parent cells (269.7±30.9) vs (62.7±10.1) and (172.0±30.8) vs (36.3±4.9), with statistical significance (all P<0.05). Conclusion:Concentration gradient increment method can successfully establish gemcitabine-resistant pancreatic cancer cell lines, which have stronger proliferation, migration and invasiveness, and can be used to study the mechanism of drug resistance in pancreatic cancer.

Objective:To construct the gemcitabine resistant cell lines of human pancreatic cancer cell line (PANC1) and mouse pancreatic cancer cell line (PANC02), and to investigate their biological behavior changes.Methods:Gemcitabine-resistant cell lines PANC1-GR of human pancreatic cancer and PANC02-GR of mouse pancreatic cancer were induced by concentration gradient increment method. Cell count assay (CCK-8), flow cytometry, cell scratch assay and Transwell assay were used to detect the drug resistance, proliferation, cell cycle, migration and invasion of the four groups of cell lines. The drug-resistant cells were also compared with the parent cells.Results:The resistance indices of PANC1-GR and PANC02-GR were 153.3 and 185.4, respectively. The results of CCK-8 showed that with the increase of gemcitabine concentration, the proliferation of resistant cells changed significantly compared with parental cells, the population doubling time of PANC1-GR was significantly shorter than that of PANC1 (1.5±0.1) d vs (2.4±0.2) d ( t=8.00, P<0.001). The proportion of cells in S and G2/M phase increased, and the proportion of cells in G0/G1 phase decreased. The cell scratch and Transwell experiments indicated that the 24h mobility of PANC1-GR and PANC02-GR was higher than that of parent cells (47.6±2.4)% vs (28.7±6.3)% and (53.6±3.2)% vs (30.1±1.4)%, the number of individual field (200 times magnification) penetrating membrane cells was also higher than that of parent cells (269.7±30.9) vs (62.7±10.1) and (172.0±30.8) vs (36.3±4.9), with statistical significance (all P<0.05). Conclusion:Concentration gradient increment method can successfully establish gemcitabine-resistant pancreatic cancer cell lines, which have stronger proliferation, migration and invasiveness, and can be used to study the mechanism of drug resistance in pancreatic cancer.

Objective To investigate the effects of sulforaphane(SFN)in regulating the macrophage glycolysis via the arachidonate 5-lipoxygenase(ALOX5)/nuclear factor kappa B(NF-κB)signaling pathway on the progression of diabetic nephropathy(DN).Methods Bioinformatics analysis was used to identify the target genes of SFN in the treatment of DN.Human proximal tubular epithelial cell line(HK-2 cells)was induced with 30 mmol/L high glucose(HG)to create an in vitro model of DN.HK-2 cells were divided into the following groups:normal glucose(NG)group,HG group,HG+SFN(3 mmol/L)group,HG+ALOX5 group,HG+SFN(3 mmol/L)+ALOX5 group,HG-treated macrophages+HK-2 group,HG+SFN(3 mmol/L)treated macrophages s+HK-2 group,HG+ALOX5 transfection treated macrophages+HK-2 group,HG+SFN(3 mmol/L)+ALOX5 transfection treated macrophages+HK-2 group.CCK-8 assay was used to detect cell viability,Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)method was used to detect cell apoptosis;glucose and lactate levels in the cells were measured using assay kits;Western blot was performed to detect the expression of ALOX5,NF-κB,and glycolysis-related proteins hexokinase-2(HK2),pyruvate kinase M2(PKM2),glucose transporter 1(GLUT1)in each group.Diabetic nephropathy(DN)mouse models were established using streptozotocin(STZ)and treated with SFN(0.5 mg/kg).Various biochemical parameters were measured in the mice,and kidney tissue pathology was examined using H&E staining.Western blot was used to detect the expression of glycolysis-related proteins(HK2,PKM2,GLUT1)in kidney macrophages.Results Bioinformatics analysis revealed ALOX5 as the target gene of SFN in treating DN.Compared to the HG group,SFN treatment enhanced HK-2 cell viability and in-hibited apoptosis(P<0.05);concurrently,SFN treatment suppressed HG-induced macrophage glycolysis-related protein and attenuated macrophage-mediated HK-2 cellular injury(P<0.05).Western blot results showed that SFN inhibited the expression of ALOX5 and NF-κB(P<0.05).The mouse experiment results showed that SFN treatment improved kidney function and pathological changes in the kidney of DN mice,and inhibited the related protein expression of acrophage glycolysis in kidney tissue(P<0.05).Conclusion SFN improves the progression of DN by inhibiting the expression of macrophage glycolysis-related protein through the ALOX5/NF-κB signaling pathway.

Objective To explore the relationship between serum thrombospondin-1(TSP-1),cathepsin S(Cat S),and Visfatin in patients with acute myocardial infarction(AMI)and myocardial microcirculation dis-orders after PCI and their clinical prognostic value.Methods A total of 90 AMI patients who underwent PCI treatment in the hospital from June 2021 to June 2023 were enrolled in the study.They were grouped into a microcirculation disorder group(62 cases)and a normal group(28 cases)based on their myocardial microcir-culation status.According to their postoperative prognosis,they were grouped into a good prognosis group(50 cases)and a poor prognosis group(40 cases).The risk factors of poor prognosis were analyzed by Logistic re-gression,and the predictive value of serum TSP-1,Cat S and Visfatin was analyzed by receiver operating char-acteristic(ROC)curve.Results The serum levels of TSP-1,Cat S,and Visfatin in the microcirculation disor-ders group were higher than those in the normal group,and those in the poor prognosis group were higher than those in the good prognosis group,and the differences were statistically significant(P＜0.05).Elevated levels of serum TSP-1,Cat S,and Visfatin were risk factors for poor prognosis in AMI patients after PCI(P＜0.05).The efficacy of combined detection of serum TSP-1,Cat S,and Visfatin levels for predicting the prog-nosis of AMI patients after PCI was higher than that of single detection(Zcombinedprediction-TSP-1=2.245,P=0.025,Z combined prediction-Cat S=2.101,P=0.036,Z combined prediction-Visfatin=2.252,P=0.024).Conclusion The serum levels of TSP-1,Cat S and Visfatin are obviously increased in AMI patients with myocardial microcirculation disorders after PCI,and the combination of the three has relatively high efficacy in predicting the prognosis of AMI pa-tients after PCI.

Purpose To detect the expression of PP1A and GSDME and the abundance of CD8+T lymphocytes in colorectal cancer,and to explore the correlation and clinical significance of PP1A and GSDME mediated pyroptosis.Methods GEPIA da-tabase was applied to analyze the mRNA expression of PP1A and GSDME in colorectal cancer and normal tissues.Western blot assay was used to detect the expression of PP1A in colorectal cancer and the corresponding normal mucosa.Immunohisto-chemistry was applied to detect the expression of PP1A and GS-DME and CD8+T lymphocytes abundance in 107 colorectal car-cinomas and normal mucosa adjacent to the carcinomas.Spearman rank correlation was used to analyze the correlation between PP1A,GSDME and CD8+T cells abundance.Results The GEPIA database search showed that mRNA expression of PP1A and GSDME in colorectal cancer differed compared to normal tissues(P＜0.05).Western blot analysis showed that the relative expression of PP1A in colorectal cancer tissue was significantly higher than that in para-cancerous tissues(0.937 vs 0.643,P＜0.001).Immunohistochemical results showed that the expression of PP1A in colorectal cancer tissues was signifi-cantly higher than that in normal mucosa(P＜0.05).The ex-pression of GSDME in cancer tissue was closely correlated with patients'age,clinical stage and mismatch repair proteins(P＜0.05),and the distribution of CD8+T cells in the cancer infil-tration front was significantly higher than that in the normal mu-cosa,and the distribution of CD8+T cells in the cancer was cor-related with pT stage,clinical stage and lymph node metastasis.Spearman correlation analysis showed that PP1A was negatively correlated with GSDME expression(r=-0.196,P＜0.05).The overall survival PP1A-positive colorectal cancer patients was worse than that of PP1A-negative patients(P＜0.05),and the prognosis of patients was correlated with the degree of differentia-tion,lymph node metastasis,pT stage and clinical stage.Posi-tive expression of PP1A,degree of differentiation,clinical stage,pT stage and lymph node metastasis are independent risk factors affecting the prognosis of colorectal cancer patients.Conclusion PP1A is highly expressed in colorectal cancer and negatively correlated with GSDME-mediated cell pyroptosis,and the differ-ential expression of both is closely related to the progression and prognosis of colorectal cancer,which can be used as a potential indicator for judgment of the prognosis of colorectal cancer pa-tients.The differential distribution of CD8+T cells may be asso-ciated with GSDME-mediated cell pyroptosis and tumor develop-ment.

Purpose To investigate the clinicopathological significance and prognostic value of oral cancer overexpressed 1(ORAOV1)in esophageal squamous cell carcinoma(ESCC).Methods The mRNA expression of ORAOV1,ERK1 and ERK2 in 140 ESCC and 39 esophageal squamous intraepithelial neoplasia(ESIN)and 140 adjacent normal mucosal tissues were detected by RT-PCR.The relationship between their overexpres-sion and clinicopathological features was analyzed in ESCC pa-tients.Spearman correlation analysis was used to explore the correlation between ORAOV1 and ERK1 and ERK2.Results The expression level of ORAOV1 mRNA(13.158±2.246)with ESCC was significantly higher than that of low-grade ESIN(4.974±1.317),high-grade ESIN(7.398±1.891)and adja-cent tissues(1.000±0).The differences were significant(P＜0.01).High expression of ORAOV1 mRNA was positively cor-related with ESCC clinical stage and lymph node metastasis(all P＜0.05).Spearman analysis showed that the expression level of ORAOV1 mRNA was positively correlated with the expression levels of ERK1 mRNA and ERK2 mRNA in ESCC.Kaplan-Meier survival curve analysis showed that the overall survival of ESCC patients with high expression of ORAOV1 was significantly lower than that of patients with low expression,and the differ-ence was significant(P＜0.05).Conclusion The expression level of ORAOV1 gradually increases during the occurrence and development of ESCC,which is an independent prognostic factor of ESCC and is expected to become a new molecular target for early diagnosis and treatment of ESCC.

Objective:To evaluate the distribution characteristics of choroidal vascularity index (CVI) in macula among normal children using swept-source optical coherence tomography (SS-OCT), and to investigate the influencing factors.Methods:A cross-sectional study was conducted.Sixty-three children aged 6 to 12 years were enrolled in The First Affiliated Hospital of Zhengzhou University from May 2021 to November 2021.Spherical equivalent refraction, axial length (AL) and other ocular biological parameters were measured.Macula-centered CVI and choroidal thickness (ChT) were measured by SS-OCT angiography.According to the ETDRS partition, the obtained image was divided into macular central fovea (0-1 mm diameter), inner ring (1-3 mm diameter) and outer ring (3-6 mm diameter) zones.Data from the right eye were selected for statistical analysis.CVI in the three rings and four zones (superior, temporal, inferior and nasal zones) of the inner and outer rings were compared.Univariate and multiple linear regression analyses were used to analyze the correlation between CVI and sex, AL, anterior chamber depth (ACD), ChT and pupil diameter.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2021-KY-0399-003). Written informed consent was obtained from each guardian.Results:The average CVI in the macular central fovea, inner ring and outer ring were 0.35±0.12, 0.32±0.10 and 0.27±0.08, respectively, with a significant difference ( F=10.96, P<0.001), and significant differences in CVI were found in pairwise comparisons of the three ring zones (all at P<0.05). Significant differences in CVI were found among superior, temporal, inferior and nasal zones in inner and outer rings ( Fring=11.60, P=0.001; Fzone=12.02, P<0.05). The CVI was smaller in superior and nasal zones in inner ring than in temporal zone, greater in superior, temporal and inferior zones in outer ring than in nasal zone, smaller in superior zone of outer ring than that of the inner ring, and the differences were statistically significant (all at P<0.001). The single factor linear regression analysis showed that ChT in the fovea, inner ring and outer ring were the influencing factors of CVI in the three ring zones (all at β=0.001, P<0.001). Pupil diameter ( β=0.034, P=0.038; β=0.040, P=0.003; β=0.024, P=0.011) and ACD ( β=0.097, P=0.034; β=0.097, P=0.013; β=0.061, P=0.032) were the influencing factors of CVI in fovea, inner ring and outer ring.After multiple linear regression analysis, the regression equations were established as follows: CVI in the macular fovea=0.001×ChT in the macular fovea + 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring-0.301 ( R2=0.514, F=6.875, P<0.001); CVI in the inner ring=0.001×ChT in the macular fovea+ 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring+ 0.088×AL-0.307 ( R2=0.603, F=9.870, P<0.001); CVI in the outer ring=0.001×ChT in the macular fovea+ 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring-0.135 ( R2=0.601, F=9.781, P<0.001). Conclusions:In children aged 6-12 years old, the CVI is higher in the macular central fovea than in inner and outer rings, and the CVI in nasal zone is the smallest in both inner and outer rings.The thicker the ChT, the higher the CVI in all zones in the macular area; the deeper the anterior chamber, the higher the CVI in the inner ring.

Objective:To explore the application effect of psychosomatic comfort based on double C theory in improving the psychological state, comfort level and sleep quality of patients after choledocholithiasis surgery.Methods:Using the convenient sampling method, a total of 120 patients with choledocholithiasis who underwent surgical treatment in Department of General Surgery in Fuyang People's Hospital of Anhui Province were selected as the research subjects from March 2020 to December 2021. The patients were divided into the observation group and the control group by odd-even grouping method, with 60 cases in each group. The control group was given routine nursing intervention mode, while the observation group was given the psychosomatic comfort model based on double C theory. The intervention effects of the two groups were compared by using Inpatient Psychological Scale (IPS) and the Chinese version of General Comfort Questionnaire (GCQ) .Results:Before intervention, there was no statistically significant difference in IPS and GCQ scores between the two groups ( P>0.05) . After the intervention, the IPS score of the observation group was lower than those of the control group, and the difference was statistically significant ( P<0.05) . The GCQ score of the observation group was higher than that of the control group, and the difference was statistically significant ( P<0.05) . Conclusions:The psychosomatic comfort based on double C theory is helpful to improve the postoperative psychological state of patients with choledocholithiasis and improve the comfort of them.