[Objective] To analyze the prognostic value of prostate cancer (PCa) pyroptosis-related genes (PRGs) using gene expression databases and to explore the regulatory mechanism of nucleotidebinding oligomerization domain-like receptor containing pyrin domain 1 (NLRP1) in the pyroptosis of PCa cells. [Methods] Fragments per kilobase of exon model per million reads mapped (FPKM) data and clinical information from PCa and adjacent tissues from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were obtained. Differentially expressed PRGs between PCa and adjacent tissues, classified subtypes and plotted survival curves were analyzed. Univariate Cox regression analysis, least absolute shrinkage and selection operator (LASSO) regression analysis were conducted to screen prognosis-related PRGs, risk scores were calculated, and a prognostic risk model was constructed and validated. Patients were divided into high and low risk groups based on the median risk scores from the training and validation sets, and gene ontology (GO) enrichment and kyoto encyclopedia of genes and genomes (KEGG) analysis were conducted on differentially expressed PRGs. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of NLRP1 in PCa cell lines, and pyroptosis was induced in DU145 and LNCaP cells while morphological changes were observed. Western blot (WB) was performed to detect the expression of pyroptosis-related molecules. [Results] A total of 6 prognostic-related PRGs were obtained, including CHMP4C, CYCS, GPX4, GSDMB, NLRP1, and PLCG1. The risk score was positively correlated with the risk of recurrence but negatively correlated with the progression-free survival (P<0.001). The area under the receiver operating characteristic curves (AUCs) for the training set at 1, 3, and 5 years were 0.769 (95%CI: 0.652-0.878), 0.804 (95%CI: 0.736-0.882), and 0.772 (95%CI: 0.631-0.905), respectively, while those for the validation set were 0.731 (95%CI: 0.647-0.826), 0.753 (95%CI: 0.674-0.818), and 0.763 (95%CI: 0.626-0.849), respectively. Differences in expression levels of the 6 PRGs were observed between the high and low risk groups in both the training and validation sets (P<0.05). Cox regression analysis showed that T stage, prostate specific antigen (PSA), Gleason grade, and risk score were independent predictors of PCa prognosis (P<0.05). Differences in risk scores were observed among patients of different ages, T stages, and Gleason grades (P<0.05). NLRP1 was found to be lowly expressed in PCa cell lines and was involved in the regulation of pyroptosis in DU145 and LNCaP cells. [Conclusion] The prognostic risk model constructed based on PRGs has a certain predictability for the prognosis of PCa patients, and NLRP1 may be involved in the regulation of pyroptosis in PCa cells.

Objective:To investigate the changes in hepatic phase II detoxification enzymes and their mechanism in metabolic associated steatohepatitis (MASH) induced by a methionine-choline-deficient (MCD) diet in mice.Methods:Ten C57BL/6J mice were randomly divided into two groups, with five mice in each group, and fed with a control diet (NCD group) and a methionine-choline-deficient diet (MCD group) for four consecutive weeks to establish the MASH model in mice. Mice body weight was recorded weekly. Mice peripheral blood and liver tissue samples were collected after four weeks. The liver histopathological changes were observed by hematoxylin-eosin staining and Sirius red staining in liver tissue. The levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides were measured by an automatic biochemical analyzer. Triglyceride and total cholesterol were used to evaluate the lipid accumulation condition in the liver of mice with Oil red O staining. Real-time fluorescence quantitative PCR was used to detect the expression of liver inflammatory factors interleukin (IL)-1β and monocyte chemoattractant protein-1 (MCP-1) condition. Transcriptome sequencing and bioinformatics were used to analyze the changes in gene expression profiles in the liver of mice and screen differentially expressed genes. The expression conditions of phase Ⅱ detoxification enzymes glutathione S-transferase mu 4 (GSTM4), dihydronicotinamide riboside:quinone oxidoreductases (NQO-2), sulfotransferase 1β1 (SULT1β1), and uridine diphosphate glucuronosyltransferase 2 family, polypeptide A3(UGT2A3) were verified by real-time fluorescent quantitative PCR. Plasma malondialdehyde content, total antioxidant capacity (T-AOC), plasma and liver glutathione content were determined using commercial kits. The expression of nuclear factor E2-related factor 2 (Nrf2), GSTM4, and UGT1A6 was examined by Western blotting. The independent sample t-test was used for comparison between the groups. Results:The body weight of mice in the MCD group showed a gradual downward trend, while the body weight of mice in the NCD group did not change significantly following four weeks of different dietary feeding. The MCD group mice liver had yellow-white appearance with round edges. The liver/body mass index was significantly lower in the NCD group ( t=3.216, P<0.01). Hematoxylin-eosin staining showed that hepatocytes in the MCD group had an occurrence of fatty degeneration accompanied by inflammatory cell infiltration, with a higher NAFLD activity score (NAS) compared to the NCD group ( t=7.155, P<0.001). Sirius red staining showed that the the liver of the MCD group had mildly increased periportal fibers. Plasma biochemical tests indicated that plasma ALT, AST, and triglyceride levels were significantly higher in the MCD group than those in the NCD group ( t=8.920, P<0.001; t=6.696, P<0.001; t=3.904, P<0.01). Oil red O staining showed that a large number of lipid droplets accumulated in the liver tissue of the MCD group and were more severe than those in the NCD group ( t=7.405, P<0.001). The triglyceride content was significantly higher in the liver of the mice in the MCD group than that in the NCD group ( t=3.559, P<0.01), and the expression of inflammatory factors IL-1β and MCP-1 was significantly increased ( t=2.562 and 2.391, respectively, P<0.05). Transcriptome sequencing analysis showed that the expression profile of genes related to lipid metabolism was changed in the liver tissue of the mice in the MCD group. The expression of multiple phase Ⅱ detoxification enzymes was significantly downregulated. Real-time fluorescence quantitative PCR verification demonstrated that the expression of four phase Ⅱ detoxification enzymes GSTM4, NQO2, SUIL1β1, and UGT2A3 were significantly lower in the liver of the mice in the MCD group than those in the NCD group ( t=2.498, 3.570, 3.768, and 4.166, respectively, P<0.05). The detection kit showed that compared with the NCD group, the malondialdehyde content in the liver of mice in the MCD group increased ( t=3.601, P<0.01), while the plasma total glutathione ( t=11.93, P<0.001) and reduced glutathione levels were significantly reduced ( t=3.635, P<0.01). The total antioxidant capacity of the liver decreased ( t=2.872, P<0.05), and the total glutathione and reduced glutathione levels in the liver were significantly increased ( t=3.175 and 3.064, P<0.05). Western blotting showed that the expression of Nrf2, GSTM4, and UGT1A6 proteins was significantly lower in the MCD group than that in the NCD group ( t=3.385, 2.990, 2.168, P<0.05). Conclusions:The expressions of multiple phase Ⅱ detoxification enzymes and antioxidant capacity are reduced in the liver of MASH mice induced by the MCD diet, and its mechanism is related to the down-regulation of the expression of the upstream regulatory factor Nrf2 protein.

Objective To explore the role of neogambogic acid in the characteristics of colorectal cancer stem cells (CRC-CSCs) through the Wnt/β-catenin signaling pathway. Methods The colorectal cells SW480 and HCT166 were divided into control group and neogambogic acid groups (1.5, 3, 6, and 12 μmol/L). The viability of CRC-CSCs was determined by MTT method, and spheroid and clone formation assays were used to assess the capacity of spheroid formation and self-renewal ability of the cells. The effects of neogambogic acid on the apoptosis and cell cycle of CRC-CSCs were evaluated by flow cytometry assays. Real-time quantitative PCR was used to detect the mRNA expression levels of relative markers (CD133, CD44, ALDH1, Oct4, and Nanog) of CRC-CSCs, and the protein expression levels of the self-renewal marker (PCNA), apoptosis markers (cleaved caspase-3 and cleaved caspase-9), and Wnt/β-catenin signaling pathway markers (p-GSK3β, GSK3β, β-catenin, and Wnt) were analyzed using Western blot. Results Compared with the control group, after neogambogic acid treatment, the viability of SW480 and HCT116 cells decreased (P<0.05), the spheroid forming ability and the clone numbers of CRC-CSCs decreased (P<0.001, P<0.01) but the cell apoptosis rate increased (P<0.01), and cell cycle was arrested in G₀/G₁ phase. Moreover, neogambogic acid downregulated the mRNA and protein expression of relative markers of CRC-CSCs (CD133, CD44, ALDH1, Oct4, and Nanog), PCNA, p-GSK3β, β-catenin, and Wnt (P<0.05) and upregulated the expression of cleaved caspase-3, cleaved caspase-9, and GSK3β (P<0.01). Conclusion Neogambogic can inhibit the stem cell properties of colorectal cells via inhibition of Wnt/β-catenin signaling pathway. As a result, neogambogic acid may be an attractive agent against colorectal cancer.

Crohn's disease (CD), a chronic and recurrent inflammatory bowel disease, requires repeated intestinal evaluations. As a non-radioactive, non-invasive, well-tolerated, inexpensive and easily reproducible detection tool, intestinal ultrasound (IUS) has been more and more widely used in the diagnosis and treatment of CD in recent years. Various parameters of IUS, such as bowel wall thickness, bowel wall stratification, color doppler signals, and inflammatory mesenteric fat, provide a lot of critical information in the diagnosis and treatment of CD. IUS can not only accurately diagnose CD and its complications, but also well evaluate CD disease activity and treatment response, and effectively predict CD transmural remission, disease duration, surgical risk, and postoperative recurrence. IUS has demonstrated good accuracy in the diagnosis, evaluation and prediction of CD.

Objective:To investigate the association of regulatory T cell (Treg) levels in peripheral blood with bronchopulmonary dysplasia (BPD) in preterm infants and its predictive value for BPD.Methods:In this case-control study, a total of 102 infants with gestational age ≤32 weeks who were hospitalized in the Neonatal Intensive Care Unit of the First Affiliated Hospital of Xinxiang Medical University from April 2022 to April 2024 were included.They were divided into a BPD group (31 cases) and a non-BPD group (71 cases) based on the diagnostic criteria of BPD.Peripheral blood samples were collected on 0 day, 7 days, 14 days, 21 days and 28 days after birth.Differences in Treg levels between the 2 groups and the relationship between Treg levels and BPD were analyzed.The independent sample t test or χ2 test was used to analyze differences between the 2 groups.One-Way ANOVA was used to compare data between groups.Multivariate Logistic regression was used to analyze the risk factors of BPD.The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of Treg levels on 7 days after birth for early diagnosis of BPD. Results:Gestational age[(28.1±1.4) weeks vs.(30.9±1.0) weeks], birth weight[(1 024±243) g vs.(1 301±188) g], Apgar score at 1 minute after birth[(4.3±1.9) points vs.(7.8±1.9) points], Apgar score at 5 minutes after birth[(7.2±1.7) points vs.(9.1±1.3) points], proportion of invasive mechanical ventilation time ≥7 days [87.1%(27/31) vs.45.1%(32/71)] and oxygen inhalation time[(45.1±11.7) days vs.(19.7±7.3) days] were statistically significantly different between BPD and non-BPD groups (all P<0.05).The Treg level in the peripheral blood of preterm infants increased first and then decreased after birth, with the peak observed on 7 days after birth.On 7 days after birth, the BPD group had a significantly higher Treg level than the non-BPD group[(10.4±1.2)% vs.(8.7±1.7)%] ( P<0.05).The multivariate Logistic regression analysis showed increased Treg levels in peripheral blood on 7 days after birth ( OR=3.320, 95% CI: 1.057-10.427, P=0.040), gestational age ( OR=0.040, 95% CI: 0.003-0.446, P=0.009), invasive mechanical ventilation time ≥7 days ( OR=4.126, 95% CI: 1.301-14.125, P=0.002), and oxygen inhalation time ( OR=1.716, 95% CI: 1.317-3.933, P=0.041) were risk factors of BPD in preterm infants.The ROC curve analysis showed that the area under the curve of Treg levels on 7 days after birth for BPD prediction was 0.794, the best cut-off value was 9.35%, the sensitivity was 90.3%, and the specificity was 66.2%. Conclusions:Treg levels in the peripheral blood of preterm infants increase first and then decrease in the early stage after birth, peaking at 7 days after birth.Elevated Treg levels at 7 days after birth may have early predictive value for the occurrence of BPD in preterm infants.

Crohn's disease (CD), a chronic and recurrent inflammatory bowel disease, requires repeated intestinal evaluations. As a non-radioactive, non-invasive, well-tolerated, inexpensive and easily reproducible detection tool, intestinal ultrasound (IUS) has been more and more widely used in the diagnosis and treatment of CD in recent years. Various parameters of IUS, such as bowel wall thickness, bowel wall stratification, color doppler signals, and inflammatory mesenteric fat, provide a lot of critical information in the diagnosis and treatment of CD. IUS can not only accurately diagnose CD and its complications, but also well evaluate CD disease activity and treatment response, and effectively predict CD transmural remission, disease duration, surgical risk, and postoperative recurrence. IUS has demonstrated good accuracy in the diagnosis, evaluation and prediction of CD.

Objective:To investigate the association of regulatory T cell (Treg) levels in peripheral blood with bronchopulmonary dysplasia (BPD) in preterm infants and its predictive value for BPD.Methods:In this case-control study, a total of 102 infants with gestational age ≤32 weeks who were hospitalized in the Neonatal Intensive Care Unit of the First Affiliated Hospital of Xinxiang Medical University from April 2022 to April 2024 were included.They were divided into a BPD group (31 cases) and a non-BPD group (71 cases) based on the diagnostic criteria of BPD.Peripheral blood samples were collected on 0 day, 7 days, 14 days, 21 days and 28 days after birth.Differences in Treg levels between the 2 groups and the relationship between Treg levels and BPD were analyzed.The independent sample t test or χ2 test was used to analyze differences between the 2 groups.One-Way ANOVA was used to compare data between groups.Multivariate Logistic regression was used to analyze the risk factors of BPD.The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of Treg levels on 7 days after birth for early diagnosis of BPD. Results:Gestational age[(28.1±1.4) weeks vs.(30.9±1.0) weeks], birth weight[(1 024±243) g vs.(1 301±188) g], Apgar score at 1 minute after birth[(4.3±1.9) points vs.(7.8±1.9) points], Apgar score at 5 minutes after birth[(7.2±1.7) points vs.(9.1±1.3) points], proportion of invasive mechanical ventilation time ≥7 days [87.1%(27/31) vs.45.1%(32/71)] and oxygen inhalation time[(45.1±11.7) days vs.(19.7±7.3) days] were statistically significantly different between BPD and non-BPD groups (all P<0.05).The Treg level in the peripheral blood of preterm infants increased first and then decreased after birth, with the peak observed on 7 days after birth.On 7 days after birth, the BPD group had a significantly higher Treg level than the non-BPD group[(10.4±1.2)% vs.(8.7±1.7)%] ( P<0.05).The multivariate Logistic regression analysis showed increased Treg levels in peripheral blood on 7 days after birth ( OR=3.320, 95% CI: 1.057-10.427, P=0.040), gestational age ( OR=0.040, 95% CI: 0.003-0.446, P=0.009), invasive mechanical ventilation time ≥7 days ( OR=4.126, 95% CI: 1.301-14.125, P=0.002), and oxygen inhalation time ( OR=1.716, 95% CI: 1.317-3.933, P=0.041) were risk factors of BPD in preterm infants.The ROC curve analysis showed that the area under the curve of Treg levels on 7 days after birth for BPD prediction was 0.794, the best cut-off value was 9.35%, the sensitivity was 90.3%, and the specificity was 66.2%. Conclusions:Treg levels in the peripheral blood of preterm infants increase first and then decrease in the early stage after birth, peaking at 7 days after birth.Elevated Treg levels at 7 days after birth may have early predictive value for the occurrence of BPD in preterm infants.

Objective:To investigate the changes in hepatic phase II detoxification enzymes and their mechanism in metabolic associated steatohepatitis (MASH) induced by a methionine-choline-deficient (MCD) diet in mice.Methods:Ten C57BL/6J mice were randomly divided into two groups, with five mice in each group, and fed with a control diet (NCD group) and a methionine-choline-deficient diet (MCD group) for four consecutive weeks to establish the MASH model in mice. Mice body weight was recorded weekly. Mice peripheral blood and liver tissue samples were collected after four weeks. The liver histopathological changes were observed by hematoxylin-eosin staining and Sirius red staining in liver tissue. The levels of plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST) and triglycerides were measured by an automatic biochemical analyzer. Triglyceride and total cholesterol were used to evaluate the lipid accumulation condition in the liver of mice with Oil red O staining. Real-time fluorescence quantitative PCR was used to detect the expression of liver inflammatory factors interleukin (IL)-1β and monocyte chemoattractant protein-1 (MCP-1) condition. Transcriptome sequencing and bioinformatics were used to analyze the changes in gene expression profiles in the liver of mice and screen differentially expressed genes. The expression conditions of phase Ⅱ detoxification enzymes glutathione S-transferase mu 4 (GSTM4), dihydronicotinamide riboside:quinone oxidoreductases (NQO-2), sulfotransferase 1β1 (SULT1β1), and uridine diphosphate glucuronosyltransferase 2 family, polypeptide A3(UGT2A3) were verified by real-time fluorescent quantitative PCR. Plasma malondialdehyde content, total antioxidant capacity (T-AOC), plasma and liver glutathione content were determined using commercial kits. The expression of nuclear factor E2-related factor 2 (Nrf2), GSTM4, and UGT1A6 was examined by Western blotting. The independent sample t-test was used for comparison between the groups. Results:The body weight of mice in the MCD group showed a gradual downward trend, while the body weight of mice in the NCD group did not change significantly following four weeks of different dietary feeding. The MCD group mice liver had yellow-white appearance with round edges. The liver/body mass index was significantly lower in the NCD group ( t=3.216, P<0.01). Hematoxylin-eosin staining showed that hepatocytes in the MCD group had an occurrence of fatty degeneration accompanied by inflammatory cell infiltration, with a higher NAFLD activity score (NAS) compared to the NCD group ( t=7.155, P<0.001). Sirius red staining showed that the the liver of the MCD group had mildly increased periportal fibers. Plasma biochemical tests indicated that plasma ALT, AST, and triglyceride levels were significantly higher in the MCD group than those in the NCD group ( t=8.920, P<0.001; t=6.696, P<0.001; t=3.904, P<0.01). Oil red O staining showed that a large number of lipid droplets accumulated in the liver tissue of the MCD group and were more severe than those in the NCD group ( t=7.405, P<0.001). The triglyceride content was significantly higher in the liver of the mice in the MCD group than that in the NCD group ( t=3.559, P<0.01), and the expression of inflammatory factors IL-1β and MCP-1 was significantly increased ( t=2.562 and 2.391, respectively, P<0.05). Transcriptome sequencing analysis showed that the expression profile of genes related to lipid metabolism was changed in the liver tissue of the mice in the MCD group. The expression of multiple phase Ⅱ detoxification enzymes was significantly downregulated. Real-time fluorescence quantitative PCR verification demonstrated that the expression of four phase Ⅱ detoxification enzymes GSTM4, NQO2, SUIL1β1, and UGT2A3 were significantly lower in the liver of the mice in the MCD group than those in the NCD group ( t=2.498, 3.570, 3.768, and 4.166, respectively, P<0.05). The detection kit showed that compared with the NCD group, the malondialdehyde content in the liver of mice in the MCD group increased ( t=3.601, P<0.01), while the plasma total glutathione ( t=11.93, P<0.001) and reduced glutathione levels were significantly reduced ( t=3.635, P<0.01). The total antioxidant capacity of the liver decreased ( t=2.872, P<0.05), and the total glutathione and reduced glutathione levels in the liver were significantly increased ( t=3.175 and 3.064, P<0.05). Western blotting showed that the expression of Nrf2, GSTM4, and UGT1A6 proteins was significantly lower in the MCD group than that in the NCD group ( t=3.385, 2.990, 2.168, P<0.05). Conclusions:The expressions of multiple phase Ⅱ detoxification enzymes and antioxidant capacity are reduced in the liver of MASH mice induced by the MCD diet, and its mechanism is related to the down-regulation of the expression of the upstream regulatory factor Nrf2 protein.

Objective:To study the influence of electromyographic biofeedback therapy(EMGBFT)combined with mirror therapy(MT)on lower limb motor and balance function in stroke patients. Method:Sixty patients with hemiplegia after stroke were randomly divided into two groups:MT based EMG-BFT group and EMGBFT group,30 patients in each group.On the basis of conventional rehabilitation,the patients in the EMGBFT group received sham MT stimulation combined with EMGBFT,and the patients in the MT based EMGBFT group received MT combined with EMGBFT.Before and after treatment,the lower limb motor function of the patients was evaluated using Fugl-Meyer assessment scale-lower extremity(FMA-LE)and surface electromyography-integrated electromyography(iEMG)of knee flexion and ankle dorsiflex-ion,co-contraction ratio(CR).Plantar pressure-symmetry index(SI)of mean pressure and contact area of both feet,elliptical area of body center of gravity,anteroposterior(AP)and mediolateral(ML)displacement distance of body center of gravity under eye-opening and eye-closed states were calculated to evaluate pa-tients'weight-bearing and balance function. Result:After treatment,FMA-LE,CR and iEMG of biceps femoris and rectus femoris under knee flexion,tibialis anterior and medial gastrocnemius under ankle dorsiflexion were markedly ameliorated in the two groups(P＜0.01).After treatment,in the eye-opening state,the SI of mean pressure and contact area of both feet,elliptical area of body center of gravity,AP and ML displacement distances of body center of gravity were greatly enhanced in the two groups(P＜0.05,P＜0.01),in the eye-closed state,the SI of mean pressure and contact area of both feet,ML displacement distances of body center of gravity were observably ameliorat-ed in the two groups(P＜0.05,P＜0.01).Compared with the EMGBFT group,the FMA-LE,iEMG of biceps femoris and tibialis anterior muscles,elliptical area of body center of gravity,AP and ML displacement dis-tance of body center of gravity with eyes open,SI of contact area of both feet with eyes closed had more significant changes in the MT based EMGBFT group after treatment(P＜0.05,P＜0.01). Conclusion:Electromyographic biofeedback therapy combined with mirror therapy can improve lower limb motor and balance function in stroke patients,the underlying mechanism of which may be the activation of lower limb weak muscle motor units,the relief of lower limb spasm,and the improvement of standing static balance ability.

Objective:To analyze influencing factors for the clinical efficacy of Perampanel monotherapy in children with focal epilepsy and promote rational clinical use of Perampanel.Methods:Children who were diagnosed with focal epilepsy and treated with Perampanel monotherapy in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science & Technology from June 2021 to August 2022 were included in this retrospective study.Efficacy at 3, 6 and 12 months after treatment was taken as the dependent variable.The gender, age at onset, age at initiation of Perampanel monotherapy, course of disease, weight, body mass index (BMI), seizure frequency, epilepsy syndrome, etiology, previous clinical history, developmental retardation, starting and maintenance doses, blood concentration, CYP3A4 and CYP3A5 gene mutations were taken as the independent variables.The effects of these factors on the efficacy of Perampanel monotherapy were analyzed using the Cox proportional hazards regression.The random forest and decision tree models were used to sequence the significance of the influencing factors and find the optimal cut-point for classification. Results:A total of 43 children (31 boys and 12 girls) were enrolled in this study.The average age at onset was (7.6±2.1) years, and the average age at initiation of Perampanel monotherapy was (7.8±2.7) years.The Cox proportional hazards regression analysis indicated that the greater the BMI, the worse the efficacy of Perampanel ( HR=0.74, 95% CI: 0.55-0.99, P=0.045). Through machine learning, the BMI was found to be a significant covariate affecting the efficacy, and when BMI≥21.8 kg/m 2, the negative effect was more significant. Conclusions:The clinical efficacy of Perampanel monotherapy in children with focal epilepsy is related to BMI.The dose of Perampanel for obese children with epilepsy may need to appropriately increase.