Mutations in ion channel genes have long been implicated in a spectrum of epilepsy syndromes. However, therapeutic decision-making is relatively complex for epilepsies associated with channelopathy. Therefore, in the present study, we used a patient-derived organoid model with a novel SCN2A mutation (p.E512K) to investigate the potential of utilizing such a model as a platform for preclinical testing of anti-seizure compounds. The electrophysiological properties of the variant Nav1.2 exhibited gain-of-function effects with increased current amplitude and premature activation. Immunofluorescence staining of patient-derived cortical organoids (COs) displayed normal neurodevelopment. Multielectrode array (MEA) recordings of patient-derived COs showed hyperexcitability with increased spiking and remarkable network bursts. Moreover, the application of patient-derived COs for preclinical drug testing using the MEA showed that they exhibit differential responses to various anti-seizure drugs and respond well to carbamazepine. Our results demonstrate that the individualized organoids have the potential to serve as a platform for preclinical pharmacological assessment.
Organoids/physiology* ; NAV1.2 Voltage-Gated Sodium Channel/genetics* ; Humans ; Anticonvulsants/pharmacology* ; Epilepsy/drug therapy* ; Mutation ; Cerebral Cortex/drug effects* ; Action Potentials/drug effects* ; Carbamazepine/pharmacology*

Objective To investigate the expression of serum microRNA(miR)-494 and miR-155 in chil-dren with Enterovirus type 71 hand-foot-mouth disease(EV71-HFMD),and to analyze the clinical significance of serum miR-494 and miR-155 in EV71-HFMD.Methods A total of 145 children with EV71-HFMD treated in this hospital from May 2021 to May 2023 were selected.According to the severity of the disease,they were divided into mild group(n=81)and severe group(n=64).In addition,73 healthy and disease-free children in the hospital were included in the health group.Serum expression levels of miR-494 and miR-155 as well as helper T cell 17(Th17),regulatory T cell(Treg)and Th17/Treg in the three groups were detected and com-pared.Pearson correlation analysis was used to analyze the correlation between the expression levels of serum miR-494,miR-155,Th17 and Treg,as well as Th17/Treg.The clinical data of children with EV71-HFMD were collected,and the factors influencing the severity of EV71-HFMD were analyzed by multivariate Logistic regression model.Results Compared with healthy group,serum miR-494,miR-155,Th17 and Th17/Treg in mild and severe groups were increased,and Treg were decreased(P＜0.05).Compared with mild group,ser-um miR-494,miR-155,Th17 and Th17/Treg in severe group were increased,and Treg was decreased(P＜0.05).Pearson correlation analysis showed that Th17 and Th17/Treg were positively correlated with the ex-pression levels of serum miR-494 and miR-155,while Treg was negatively correlated with the expression lev-els of serum miR-494 and miR-155(P＜0.05).The peak temperature,duration of fever ≥3 d proportion and eruption of isthmus in severe group were higher than those in mild group(P＜0.05).Multivariate Logistic re-gression results showed that the duration of fever ≥3 d,eruption in the isthmus,the high expression level of miR-494 and the high expression level of miR-155 were risk factors for severe EV71-HFMD(P＜0.05).Conclusion Se-rum miR-494 and miR-155 are abnormally elevated in children with EV71-HFMD,and the changes in their levels are closely related to Th17/Treg imbalance.Increased expression levels of serum miR-494 and miR-155 are risk factors for severe EV71-HFMD.

Objective : To compare the antiviral efficacy and renal safety of nucleoside analogs(NAs) monotherapy versus combination therapy in hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB) patients with high viral load. Methods : This study enrolled a total of 353 treatment-naïve HBeAg-positive chronic hepatitis B (CHB) patients with high viral load , the treatment regimen was divided into 5 groups , consisting of 4 monotherapy groups and 1 combination therapy group as follows : 88 cases in the Entecavir (ETV) group , 135 cases in the Teno- fovir Disoproxil Fumarate (TDF) group , 34 cases in the Tenofovir Alafenamide Fumarate (TAF) group , 25 cases in the Tenofovir Amibufenamide (TMF) group , and 71 cases in the ETV combined with TDF (ETV + TDF) group . A retrospective cohort study design was adopted to analyze HBV DNA levels , serological indicators ( HBsAg and HBeAg levels) , renal function indicators ( serum Scr levels , eGFR) at 24 and 48 weeks of treatment across various groups , as well as the HBsAg clearance rates , HBeAg seroconversion rates and HBV DNA suppression rates (HBV DNA < 20 IU/ml) at 48 weeks across the groups . Multivariate logistic regression analysis was conducted to identify the influencing factors for HBV DNA suppression . Results : At 24 weeks , the HBV DNA level in the ETV + TDF selected . Key miRNAs included hsa-let-7b-5p , hsa-let-7c-5p , hsa-let-7b-3p _ 1ss22CT , and hsa-miR-199b-5p , with BACH1 and IFNAR1 identified as their shared target genes . GO analysis revealed that the enriched target genes were primarily involved in protein binding , metal ion binding , transferase activity , DNA binding , transcriptional regulation by RNA polymerase Ⅱ , and nucleotide binding. KEGG pathway analysis indicated that the target genes were mainly associated with metabolic pathways , cancer-related pathways , the PI3K-Akt signaling pathway , and the Rap1 signaling pathway . Conclusion Differential expression of miRNAs in amniotic fluid exosomes was ob- served between DS fetuses and those with normal karyotypes . Combined analysis with placental miRNAs revealed hsa-miR-199b-5p as a common differentially expressed miRNA in both DS amniotic fluid and placenta. It is hypoth- esized that BACH1 and IFNAR1 , shared target genes of hsa-miR-199b-5p , hsa-let-7b-5p , hsa-let-7c-5p , and hsa- let-7b-3p_1ss22CT , may play a role in the pathogenesis of DS .

Objective To explore the mechanism of ginseng in the treatment of periodontitis based on network phar-macology and molecular docking technology.Methods Potential targets of ginseng and periodontitis were obtained through various databases.The intersection targets of ginseng and periodontitis were obtained by using VENNY,the pro-tein-protein interaction network relationship diagram was formed on the STRING platform,the core target diagram was formed by Cytoscape software,and the ginseng-active ingredient-target network diagram was constructed.The selected targets were screened for gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway en-richment analysis.The core targets of ginseng's active in-gredients in treating periodontitis were analyzed by mo-lecular docking technique.Results The 22 ginseng's active ingredients,591 potential targets of ginseng's ac-tive ingredients,2 249 periodontitis gene targets,and 145 ginseng-periodontitis intersection targets were analyzed.Ginseng had strong binding activity on core targets such as vas-cular endothelial growth factor A and epidermal growth factor receptor,as well as hypoxia induced-factor 1(HIF-1)sig-naling pathway and phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt)signaling pathway.Conclusion Gin-seng and its active components can regulate several signaling pathways such as HIF-1 and PI3K-Akt,thereby indicating that ginseng may play a role in treating periodontitis through multiple pathways.

Objective:To investigate the spatial distribution pattern of local tumor progression (LTP) for hepatocellular carcinoma (HCC) ≤5 cm after microwave ablation.Methods:A retrospective analysis was performed on 169 HCCs with matched MRI before and after ablation from December 2009 to December 2019. A tumor MRI was reconstructed using three-dimensional visualization technology. LTP was classified as contact or non-contact, early or late stage, according to whether LTP was in contact with the edge of the ablation zone and the occurrence time (24 months). The tumor-surrounded area was divided into eight quadrants by using the eight-quadrant map method. An analysis was conducted on the spatial correlation between the quadrant where the ablative margin (AM) safety boundary was located and the quadrant where different types of LTP occurred. The t-test, or rank-sum test, was used for the measurement data. 2-test for count data was used to compare the difference between the two groups.Results:The AM quadrant had a distribution of 54.4% LTP, 64.2% early LTP stage, and 69.1% contact LTP, suggesting this quadrant was much more concentrated than the other quadrants ( P ?

Objective To investigate the protective effects and underlying mechanisms of sodium butyrate on rats exposed to hypoxia and cold conditions.Methods Fifty-eight male SD rats (aged 7～8 weeks,weighing 240～260 g ) were randomly divided into normoxia normothermia saline control (NNC ) group (n=10),normoxia normothermia sodium butyrate(NNB)group(n=10),hypoxia cold saline control (HCC) group (n=19),and hypoxia cold sodium butyrate (HCB)group (n=19).The intragastric dose of sodium butyrate was 200 mg/kg for the NNB and HCB groups,while the NNC and HCC groups were given normal saline of 5 mL/kg.①After continuous intragastric administration for 7 d,the rats in the HCC and HCB groups were placed in a low-pressure hypoxic chamber to simulate an altitude of 5000 m and exposed to a temperature of 8 ℃ for 7 d.Subsequently,blood samples were collected from the abdominal aorta for blood gas analysis,blood routine test,and detection for serum biochemical indicators.ELISA was used to determine serum inflammatory cytokines and endocrine hormones.The rats in the NNC and NNB groups(n=10)were fed outside the chamber and underwent the same tests in 7 d later to evaluate the protective effects of sodium butyrate on the body.②Core body temperature monitoring was conducted to assess the impact of sodium butyrate on the rmoregulation in rats exposed to hypoxia and cold(n=3).③Hypoxia exercise tolerance of the HCC group and HCB group in a hypoxic chamber (11％O2 )was evaluated for their hypoxia resistance (n=6).Results Compared to the NNC group,the rats in the HCC group exhibited significant decreases in arterial oxygen saturation (SaO2 )and arterial oxygen partial pressure (PaO2 ),serum levels of IL-4,estradiol (E2)and cortisol (F),core temperature,and exercise duration (P<0.05),and had notably increased levels of red blood cell (RBC)count,hemoglobin (HGB),hematocrit (HCT),cardiac troponin (CRDAC-T),uric acid (UA),alanine aminotransferase (ALT),total cholesterol (TC),low-density lipoprotein (LDL),IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF)(P<0.05).Compared to the HCC group,the rats in the HCB group exhibited significant increases in SaO2,PaO2,IL-4,E2,F,corticotropin releasing hormone (CRH)and adrenocorticotropic hormone (ACTH)(P<0.05),remarkably longer exercise duration under hypoxic exposure (P<0.05 ),but decreases in RBC count,serum levels of HGB,HCT,CRDAC-T,UA,ALT,TC,LDL,IL-6,GM-CSF and free thyroxine (FT4 ),and core temperature (P<0.05).Conclusion Sodium butyrate exhibits protective effects on rats exposed to hypoxia and cold conditions,and it is helpful in their adaptation to these hypoxia and cold environments.

Gastric cancer is among the most common malignant tumors worldwide.Due to its subtle early symptoms and suboptimal screening rates,a significant proportion of patients are diagnosed with advanced-stage gastric cancer,which adversely influences patient prognosis.In recent years,with the advancement of research,immunotherapy for gastric cancer has made significant strides,demonstrating particularly remarkable efficacy in cases of unresectable and metastatic gastric cancer.This has established it as an effective treatment method following surgery,chemotherapy,radiotherapy,and targeted therapy.Immunotherapy involves artificially modulating the immune system using strategies such as adoptive cell therapy,immune checkpoint inhibitors,and tumor vaccines.Although immunotherapy has broad prospects,it faces many challenges and problems in clinical application.This review delineates the clinical application of immunother-apy in gastric cancer and discusses the extant challenges therein.

Objective:To investigate the cognitive functions of temporal lobe epilepsy (TLE) patients with bilateral asynchronous interictal discharges.Methods:A total of 162 TLE patients who were treated at Zhongshan Hospital, Fudan University, from June 2021 to December 2023 were collected. According to the interictal scalp electroencephalogram, TLE patients were classified to the TLE with bilateral temporal asynchronous interictal epileptiform discharges ( n=51) and TLE with unilateral temporal epileptiform discharges ( n=111). Unilateral TLE patients were divided into TLE with right ( n=48) and left ( n=63) temporal epileptiform discharges. The Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Digital Span (DS), Verbal Fluency Test (VFT), the third part of Color Word Test (CWT-C), Trail Making Test-B (TMT-B), Symbol Digit Modalities Test (SDMT), Auditory Verbal Learning Test (AVLT), Rey-Osterrieth Complex Figure (ROCF), and Similarity Comprehension Test were performed for the participants. The differences of cognitive functions between patients with bilateral and unilateral temporal discharges were compared. Univariate and multivariate Logistic regression models were used to analyze the risk factors for patients with bilateral temporal asynchronous discharges. Spearman analysis was used to explore the correlation between cognitive function and clinical indicators. Results:In the group of TLE patients with bilateral asynchronous discharges compared to those with unilateral discharges, the completion time of CWT-C [67 (55, 103) s vs 59 (50, 71) s, Z=-2.904, P=0.004], TMT-B [159 (108, 219) s vs 129 (95, 180) s, Z=-2.361, P=0.018] was longer. Additionally, TLE patients with bilateral asynchronous discharges got lower scores of MMSE [28 (26, 29) vs 29 (28, 30), Z=3.098, P=0.002], MoCA [23 (19, 28) vs 27 (23, 28), Z=3.175, P=0.001], AVLT1+2+3 [16.843±6.482 vs 19.162±5.526, t=-2.347, P=0.020], AVLT6 [6 (3, 10) vs 8 (5, 10), Z=3.275, P=0.001], ROCF2 [15 (8, 22) vs 20 (12, 25), Z=2.870, P=0.004], ROCF3 [14 (8, 22) vs 20 (11, 25), Z=2.634, P=0.008], and Similarity Test [13 (8, 18) vs 16 (12, 20), Z=2.387, P=0.017] as well as lower VFT-vegetable and fruit count [15 (13, 19) vs 18 (15, 21), Z=2.402, P=0.016] and SDMT completion count [41 (30, 53) vs 51 (40, 60), Z=3.089, P=0.002]. The multivariate Logistic regression analysis showed that the decrease in AVLT6 scores ( OR=1.546, 95% CI 1.150-2.078, P=0.004) and longer TMT-B time ( OR=1.013, 95% CI 1.001-1.025, P=0.035) were independent risk factors for TLE patients with bilateral asynchronous discharges. Conclusions:Compared to the patients with TLE characterized by unilateral temporal lobe discharges, those with asynchronous discharges in bilateral temporal lobes show statistically significant declines in all domains of cognitive functions, including executive function, memory, and language abilities. Decreased cue recall ability in language memory and prolonged trail-making test in executive function are independent cognitive impairment risk factors for bilateral temporal asynchronous discharges.

OBJECTIVE To study the effects of Yishen daluo decoction on inflammatory factors and cyclic adenosine monophosphate（cAMP）/protein kinase A （PKA）/cAMP response element binding protein （CREB） signal pathway in experimental autoimmune encephalomyelitis （EAE） model mice by inhibiting the expressions of β-arrestin1， and to explore the mechanism of Yishen daluo decoction in the treatment of EAE. METHODS Sixty mice were randomly divided into normal group， model group， TCM group （Yishen daluo decoction 20 g/kg）， positive control group （prednisone acetate 3.9 mg/kg）， β-arrestin1 siRNA adeno- associated virus （AAV-β） group， AAV-β+TCM group， with 10 mice in each group. Except for normal group， EAE model was made in other groups. AAV-β group and AAV-β+TCM group were injected with AAV-β via tail vein to interfere with the expression of β -arrestin1 protein. Starting from the 8th day of modeling， they were given corresponding drug solution/normal saline intragastrically， once a day， for consecutive 14 days. The neurological function score of mice was detected； the pathological and morphological changes were observed in the brain and spinal cord tissues of mice； the serum levels of inflammatory factors [interleukin-2 （IL-2）， IL-23， interferon-γ （IFN-γ）] in mice were determined； the expressions of β-arrestin1， cAMP， PKA and CREB in brain and spinal cord were detected. RESULTS Compared with normal group， neurological function scores， serum levels of inflammatory factors， and protein expressions of β-arrestin1 in brain and spinal cord were significantly increased （P＜0.05 or P＜ 0.01）； protein expressions of PKA， CREB and cAMP in brain and spinal cord were decreased significantly（P＜0.05 or P＜0.01）. The deep staining of cellular shrinkage and aggregation of inflammatory cells were observed in most neurons of the brain and spinal cord， with varying degrees of demyelinating. Compared with model group， the neurological function scores， pathological changes in brain and spinal cord tissues， and most indicators （except for CREB and cAMP proteins in the brain tissue of AAV-β group） were significantly reversed （P＜0.05 or P＜0.01）.Compared with AAV- β group， the neurological function scores， the levels of IFN-γ in serum and β-arrestin1 in spinal cord were significantly decreased （P＜0.05 or P＜0.01）， PKA and cAMP in brain and spinal cord tissues were significantly increased in AAV- β +TCM group （P＜0.05 or P＜0.01）. CONCLUSIONS Yishen daluo decoction can inhibit the expression of β-arrestin1 in the central nervous system thus activating the cAMP/PKA/CREB signaling pathway， relieving nervous system inflammation， and ultimately alleviates the symptoms of EAE.

ObjectiveTo investigate the effects of Yishen Daluo prescription （YSDL） on Ras homolog（Rho）/Rho-associated coiled-coil containing protein kinase（ROCK）signaling pathway in mice with experimental autoimmune encephalomyelitis （EAE） based on the silencing of β-arrestin1 gene. MethodSixty C57BL/6 female mice were randomly divided into a blank group， a model group， a virus group， a YSDL group， a virus + YSDL group， and a prednisone acetate group （hormone group）. The EAE model was induced in mice except for those in the normal group. Adeno-associated virus（AAV）solution （150 μL， 1×10¹¹ vg·mL^-1） was injected into the tail vein of each mouse in the virus group and the virus + YSDL group on the 4^th day of immunization. Drugs were administered on the 8^th day of modeling. Specifically， normal saline was given to the mice in the normal group，the model group，and the virus group at 10 mL∙kg^-1， prednisone acetate suspension to those in the hormone group at 3.9 g∙kg^-1，and YSDL to those in other groups at 20 g∙kg^-1 for 14 consecutive days. The mice were weighed and scored every day. The neurological function scores of mice in each group were recorded every day after immunization. Hematoxylin-eosin （HE） staining was used to determine the inflammatory response and lesion location in the brain tissues and spinal cord tissues of mice. The protein expression of β-arrestin1，Ras homolog gene family member A（RhoA）， and Rho-associated coiled-coil forming protein kinase Ⅰ（ROCK Ⅰ） in spinal cord and brain tissues of EAE mice was determined by Western blot. ResultCompared with the model group， the virus group and the virus + YSDL group showed decreased neurological function scores （P<0.01），and the YSDL group also showed decreased neurological function scores（P<0.05）. HE results showed that there was obvious inflammatory reaction in the central nervous system （CNS） of the model group， which was alleviated to varying degrees in other groups compared with the model group. Western blot results showed that compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the spinal cord tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression levels of β-arrestin1， RhoA， and ROCKⅠ in the spinal cord tissues （P<0.01）. Compared with the blank group， the model group showed increased protein expression levels of β-arrestin1， RhoA， and ROCK Ⅰ in the brain tissues （P<0.01）. Compared with the model group， the virus group， the YSDL group， the virus + YSDL group， and the hormone group showed decreased protein expression level of β-arrestin1 in the brain tissues （P<0.01）， and the virus group and the YSDL group showed decreased protein expression levels of RhoA， and ROCKⅠ in the brain tissues （P<0.05）. Additionally， the virus + YSDL group and the hormone group showed decreased protein expression levels of RhoA and ROCKⅠ in the brain tissues （P<0.01）. ConclusionYSDL can improve the clinical symptoms of EAE mice and improve the inflammatory response of CNS. The mechanism is presumably attributed to the fact that YSDL inhibits the expression of β-arrestin1 in CNS，thereby reducing the expression of Rho/ROCK signaling pathway. Furthermore， YSDL may have a synergistic effect with the inhibition of β-arrestin1 gene expression.