ObjectiveTo observe the inhibitory effect of the Weiliuan mixture （WLAHJ） on the subcutaneous xenograft tumor of MKN-74 gastric cancer cells， and explore the potential anti-gastric cancer mechanism of WLAHJ by using transcriptomic sequencing technology to reveal related genes and pathways. Methods30 Balb/c nude mice were randomly divided into model， low-， medium-， and high-dose（15，30，45 g·kg^-1） WLAHJ and 5-FU （0.025 g·kg^-1） groups to build a subcutaneous xenograft tumor model with MKN-74 human gastric cancer cells. After modeling，each group was continuously treated with the corresponding drugs for 28 days. During the treatment period， the body weight and tumor size of the mice were observed and recorded every 2 days. At the end of the treatment， the mice were sacrificed， and required samples were collected to calculate the tumor inhibition rate of WLAHJ on the subcutaneous xenograft tumor. High-throughput transcriptomic sequencing （RNA-seq） technology was used to analyze the differentially expressed genes in the subcutaneous tumor tissues of the model group and the medium-dose WLAHJ group， thus exploring the potential mechanism of WLAHJ in gastric cancer intervention. Immunofluorescence experiments were conducted to detect the protein expression levels of glutathione peroxidase 4 （GPX4）， solute carrier family 7 member 11 （SLC7A11）， transferrin receptor protein-1 （TFR-1）， and acyl-CoA synthetase long-chain family member 4 （ACSL4） in subcutaneous xenograft tumors of each group. Cell counting kit-8（CCK-8） and colony formation assays were used to detect the viability and anti-proliferative ability of human gastric cancer AGS and MKN-74 cells at different concentrations of WLAHJ. Kits were used to detect the levels of Fe²⁺， reactive oxygen species （ROS）， malondialdehyde （MDA）， and superoxide dismutase （SOD） activity in cells. Western blot was used to detect the expression levels of GPX4， SLC7A11， TRF-1， ACSL4， spermidine/spermine N1-acetyltransferase 1 （SAT1）， arachidonic acid 15-lipoxygenase （ALOX15）， and key proteins in the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway. ResultsThe mechanism of WLAHJ in gastric cancer intervention may be related to ferroptosis and the PI3K/Akt /mTOR signaling pathway. The growth of subcutaneous xenograft tumors in nude mice of the WLAHJ and 5-FU groups（P<0.05，P<0.01）， GPX4， and SLC7A11 dropped significantly（P<0.01）， while TFR-1， ACSL4， SAT1， and ALOX15（P<0.05，P<0.01）increased significantly compared with those in the model group. The levels of ROS， Fe²⁺， and MDA increased in the WLAHJ and 5-FU groups and the proliferation of gastric cancer cells， SOD activity， the ratios of phosphorybation （p）-mTOR/mTOR， p-PI3K/PI3K， and p-Akt/Akt protein expressions（P<0.05，P<0.01）decreased compared with those in the blank group. ConclusionThe mechanism of WLAHJ in treating gastric cancer may be related to the regulation of the PI3K/ Akt /mTOR signaling pathway to intervene in ferroptosis.

Objective To explore the effect and mechanism of quercetin on osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs were divided into a blank control group(Control)and quercetin(Quercetin)low-dose group(4.8 mL/kg),medium-dose group(9.6 mL/kg),and high-dose group(19.2 mL/kg)intervened with drug-containing serum,while the positive control group was treated with osteogenic differentiation medium,respectively.The cell cycle was analysed by flow cytometry,cell proliferation was detected by MTT assay,cell activity was determined by Alkaline phosphatase(ALP)assay kit,and calcified nodule formation was observed by alizarin red staining.The expression of β-catenin and the key factors of osteogenic differentiation,runt-related transcription factor 2(RUNX2)and osteocalcin(OCN),were detected by qPCR and Western blot,respectively.Results Compared with the control group,quercetin-containing serum significantly promoted the proliferation of BMSCs(P=0.000205,P=0.000063)and enhanced the formation of calcium nodules,and increased osteogenic and ALP activity after osteogenic differentiation.The results of qPCR and Western blot showed that the quercetin group significantly up-regulated the mRNA and protein expression of β-catenin(P<0.0001),RUNX2(P<0.0001)and OCN(P<0.0001)during osteogenic differentiation.Conclusion Quercetin can effectively promote the osteogenic differentiation of BMSCs,and its mechanism is achieved by activating the Wnt/β-catenin signaling pathway and up-regulating the expression of the osteogenesis-related transcription factor RUNX2.

Objective To analyze the clinical parameters of esophageal cancer patients before and during neoadjuvant chemotherapy,as well as to explore the related factors and predictive value that affect the efficacy of neoadjuvant chemotherapy for esophageal cancer.Methods A total of 194 patients with esophageal cancer who underwent neoadjuvant chemotherapy at the First Affiliated Hospital of Guangxi Medical University from 2020 to 2023 were selected as the research subjects.The treatment process and outcomes of the patients were followed up,and they were divided into an effective group and an ineffective group according to the effi-cacy.Differences were compared in clinical parameters between two groups of patients before and during treat-ment,screen for factors that may affect efficacy,correlation analysis was conduct to explore the correlation be-tween relevant factors and the efficacy of neoadjuvant chemotherapy,and the predictive value of relevant fac-tors were analyzed using receiver operating characteristic(ROC)curve analysis.Results There was a statisti-cally significant difference(P＜0.05)in the average cycle cost,lowest WBC value,lowest PLT value,inci-dence of nausea,transaminase abnormalities,and hair loss between the two groups.Logistic regression analysis showed that average cycle cost,transaminase abnormalities,and hair loss were related factors affecting neoad-juvant chemotherapy for esophageal cancer.Spearman correlation analysis showed that the above indicators had a certain correlation with the efficacy of neoadjuvant chemotherapy for esophageal cancer.ROC curve a-nalysis showed that the area under the curve(AUC)for predicting the efficacy of neoadjuvant chemotherapy in esophageal cancer by combining transaminase abnormalities,average cycle cost,and hair loss was 0.758(95％CI:0.683-0.832)Conclusion There is a certain correlation between average cycle cost,transaminase abnormalities,and hair loss and the effectiveness of neoadjuvant chemotherapy for esophageal cancer,which has certain predictive value for the efficacy of neoadjuvant chemotherapy for esophageal cancer.

Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Objective To observe the clinical efficacy of belimumab combined with glucocorticoids in the treatment of children with systemic lupus erythematosus(SLE).Methods A total of 64 children with SLE were randomly divided into the observation group and the control group,with 32 cases in each group.The control group was treated with oral prednisone tablets combined with hydroxychloroquine sulfate tablets,while the observation group was treated with belimumab on the basis of treatment in the control group.After 6 months of treatment,the clinical efficacy,systemic lupus erythematosus disease activity index(SLEDAI)scores and laboratory indicators[complement C3,complement C4,immunoglobulin IgG(IgG),B lymphocytes,white blood cell count(WBC),erythrocyte sedimentation rate(ESR),serum C-reactive protein(CRP),24-hour urine protein quantification(24 h UP),serum creatinine(SCr)and blood urea nitrogen(BUN)]were compared between the two groups.Results After 6 months of treatment,the total effective rate was higher in the observation group than that of the control group(93.8%vs.75.0%,P＜0.05).Compared with before treatment,both groups showed lower SLEDAI scores,IgG,B lymphocytes,ESR,CRP,SCr,24 h UP and BUN after treatment,while C3,C4 levels and WBC were higher.Moreover,the improvement of all indicators was better in the observation group than that in the control group(P＜0.05).Conclusion Belimumab combined with glucocorticoids can improve the clinical efficacy,control disease activity and enhance immune function in children with SLE.

Objective To explore an epileptic seizure prediction method for patients with refractory epilepsy to improve the classification and prediction efficiency of epileptic electroencephalogram(EEG)signals.Methods The study used the long-term EEG database of patients with intractable epilepsy from Children's Hospital Boston(CHB-MIT).The EEG features of epileptic seizures and preictal periods were extracted from multiple dimensions such as EEG synchronization,complexity,and energy distribution,and then these features were input into the artificial neural network model for classification and identification,thereby achieving accurate prediction of epilepsy.The performance were optimized by adjusting the model parameters,and a comparative evaluation was conducted with existing deep learning models.Results The model proposed in this study showed an accuracy rate of 99.29％,a precision of 91.44％,a sensitivity of 96.46％,and a specificity of 99.46％.Compared with current epilepsy seizure prediction studies based on machine learning or deep learning frameworks,the model in this study improved its classification prediction capabilities and demonstrated higher prediction accuracy.Conclusion An effective prediction of epileptic seizures was achieved by manually extracting epileptic EEG features and constructing an artificial neural network model.The model demonstrated high accuracy and stability,providing reliable technique to support clinical treatment and prevention of epilepsy.

Objective:To investigate the role of WW domain containing E3 ubiquitin protein ligase 1 (WWP1) in enamel development of mice.Methods:Single-cell RNA sequencing data of incisor tissues of postnatal day 7 (P7) mice and mandibular first molar tooth germs of P3.5 mice were used to analyze the expression of Wwp1 in dental epithelial cells. Immunohistochemistry was performed to observe the distribution and expression levels of WWP1 in the epithelium of mouse incisors and mandibular first molar tooth germs. Wwp1 knockout (Wwp1 KO) mice were generated and collected with their control littermates at P1, P7, three mice per group, as well as at P14, P28, 2 months (2M), and 3M, six mice per group. The enamel volumes of molars and incisors were analyzed using micro-CT. Scanning electron microscopy was employed to examine the enamel cross-sections of Wwp1 KO and control mice. Energy dispersive spectroscopy (EDS) was used to analyze the calcium and phosphorus content of the enamel rod of incisors. Immunofluorescence was performed to detect the expression of amelogenin (AMELX) in the ameloblasts of Wwp1 KO and control mice. Additionally, LS-8 ameloblast-like epithelial cells were cultured, and Wwp1 siRNA or overexpression plasmids were transfected to knock down or overexpress WWP1. The protein levels of AMELX were then assessed by Western blotting.Results:Single-cell sequencing result showed a high Wwp1 mRNA expression level in the epithelial cells of mouse incisors and mandibular molar tooth germs. Immunohistochemistry revealed the expression of WWP1 in presecretory, secretory, transitional, and mature ameloblasts. Wwp1 KO mice exhibited enamel developmental defects. The enamel volumes of molars and incisors in Wwp1 KO mice [(0.155±0.016), (0.300±0.017) μm 3] were reduced by 23.95% ( P<0.001) and 28.31% ( P<0.001) compared with the control group [(0.203±0.062), (0.418±0.023) μm 3] respectively. Scanning electron microscopy showed disorganized enamel structures in Wwp1 KO incisors and molars. EDS results showed the weight percent of calcium in the enamel rod of incisors decreased in Wwp1 KO mice [(20.74±0.91)%] compared with the control group [(30.30±3.83)%] ( P<0.001), and the calcium-to-phosphorus ratio decreased in Wwp1 KO mice (1.93±0.01) compared with the control group (2.02±0.01) ( P<0.001). Immunofluorescence showed weaker AMELX expression in ameloblasts of mandibular first molar tooth germs from P1 and P7 Wwp1 KO mice compared with the control group ( P<0.001, P<0.001). In LS-8 cells, Wwp1 knocked-down led to a decrease of AMELX protein expression, while WWP1 overexpression resulted in an increased AMELX protein level. Conclusions:WWP1 promotes ameloblast differentiation and enamel matrix mineralization, playing a critical role in enamel formation.

Objective To explore an epileptic seizure prediction method for patients with refractory epilepsy to improve the classification and prediction efficiency of epileptic electroencephalogram(EEG)signals.Methods The study used the long-term EEG database of patients with intractable epilepsy from Children's Hospital Boston(CHB-MIT).The EEG features of epileptic seizures and preictal periods were extracted from multiple dimensions such as EEG synchronization,complexity,and energy distribution,and then these features were input into the artificial neural network model for classification and identification,thereby achieving accurate prediction of epilepsy.The performance were optimized by adjusting the model parameters,and a comparative evaluation was conducted with existing deep learning models.Results The model proposed in this study showed an accuracy rate of 99.29％,a precision of 91.44％,a sensitivity of 96.46％,and a specificity of 99.46％.Compared with current epilepsy seizure prediction studies based on machine learning or deep learning frameworks,the model in this study improved its classification prediction capabilities and demonstrated higher prediction accuracy.Conclusion An effective prediction of epileptic seizures was achieved by manually extracting epileptic EEG features and constructing an artificial neural network model.The model demonstrated high accuracy and stability,providing reliable technique to support clinical treatment and prevention of epilepsy.

Objective To observe the clinical efficacy of belimumab combined with glucocorticoids in the treatment of children with systemic lupus erythematosus(SLE).Methods A total of 64 children with SLE were randomly divided into the observation group and the control group,with 32 cases in each group.The control group was treated with oral prednisone tablets combined with hydroxychloroquine sulfate tablets,while the observation group was treated with belimumab on the basis of treatment in the control group.After 6 months of treatment,the clinical efficacy,systemic lupus erythematosus disease activity index(SLEDAI)scores and laboratory indicators[complement C3,complement C4,immunoglobulin IgG(IgG),B lymphocytes,white blood cell count(WBC),erythrocyte sedimentation rate(ESR),serum C-reactive protein(CRP),24-hour urine protein quantification(24 h UP),serum creatinine(SCr)and blood urea nitrogen(BUN)]were compared between the two groups.Results After 6 months of treatment,the total effective rate was higher in the observation group than that of the control group(93.8%vs.75.0%,P＜0.05).Compared with before treatment,both groups showed lower SLEDAI scores,IgG,B lymphocytes,ESR,CRP,SCr,24 h UP and BUN after treatment,while C3,C4 levels and WBC were higher.Moreover,the improvement of all indicators was better in the observation group than that in the control group(P＜0.05).Conclusion Belimumab combined with glucocorticoids can improve the clinical efficacy,control disease activity and enhance immune function in children with SLE.

Objective:To compare the similarities and differences of macrophage activation syndrome（MAS）combined with systemic juvenile idiopathic arthritis（sJIA）versus with juvenile onset systemic lupus erythematosus（JSLE）.Methods:The clinical data of 48 children with MAS admitted to the Department of Nephrology and Immunology in Children's Hospital of Hebei Province from May 2015 to January 2023 were retrospectively analyzed. The patients were divided into sJIA-MAS and JSLE-MAS group，and the clinical manifestations，laboratory indicators and treatment of the two groups were compared.Results:Among the 48 children（14 males and 34 females）with MAS，the average age of onset was 9.5（3.0，11.8）years. There were 28 cases（11males and 17 females）of sJIA-MAS and 20 cases（3 males and 17 females）of JSLE- MAS. All the 48 children with MAS had fever and hyperferinemia，and the fever with sJIA-MAS was mostly continued fever or remittent fever. Respiratory tract infection was the most common trigger in sJIA-MAS［15 cases（53.6%）］，and disease activity was the most common trigger in JSLE-MAS［13 cases（65.0%）］.Additionally，viral infections（EB virus and cytomegalovirus）were also one of the triggers in MAS［sJIA：7 cases（25%），JSLE：4 cases（20%）］.Compared with JSLE-MAS，the number of days with fever［15.0（12.0，21.0）days vs. 6.0（4.0，9.5）days， Z=-3.812， P=0.001］and the length of hospital stay［29.0（26.3，39.8）days vs.26.0（19.3，30.8）days， Z=-1.958， P=0.049］were longer in sJIA. Compared with JSLE-MAS，ALT［（685.32±561.67）U/L vs.（139.61±124.44）U/L， t=4.973， P=0.001］，AST［784.00（235.25，1 251.25）U/L vs.189.50（53.25，374.08）U/L， Z=-3.283， P=0.001］，CRP［11.48（3.56，28.89）mg/L vs.1.91（0.53，8.98）mg/L， Z=-3.200， P=0.001］，ferritin［32 167.0（12 384.8，65 963.8）μg/L vs.2 003.5（922.5，11 430.0）μg/L， Z=-4.130， P=0.001］，ferritin max/ESR min［1 353.35（355.75，4 342.53）vs.91.92（34.94，291.53）， Z=-4.120， P=0.001］were higher in sJIA.The decrease of CRP was greater in sJIA［80.04（45.64，143.71）mg/L vs.10.20（6.27，25.64）mg/L， Z=-4.433， P=0.001］.Compared with sJIA-MAS，peripheral white blood cell counting［4.05（2.90，7.73）×10 9/L vs.1.56（1.15，3.47）×10 9/L， Z=-3.577， P=0.001］and platelet counting［（162.68±92.19）×10 9/L vs.（110.10±72.99）×10 9/L， t=2.118， P=0.040］were lower in JSLE-MAS. Kidney involvement was more common in JSLE-MAS［10 cases（50%）vs.0 cases（0%）， χ 2=17.684， P=0.001］.There was no significant difference in the incidence of sJIA-MAS and JSLE-MAS meeting the criteria of hemophagocytic lymphohistiocytosis［6 cases（21.4%）vs.5 cases（25.0%）， χ 2=0.084， P=0.772］. Conclusion:Compared with JSLE-MAS，sJIA-MAS is more dangerous and difficult to control，while JSLE-MAS involves more organs，among which the blood system and kidney are more common.