Objective: To explore the longitudinal relationship between bullying victimization and sleep quality among rural boarding junior high school students in Hebei Province, and to investigate the chain mediated role of loneliness and rumination, so as to provide evidence for promoting sleep health in the population. Methods: A baseline survey was conducted in May, 2023 (T1) by convenient sampling method, and two rounds of longitudinal surveys were conducted in November, 2023 (T2) and May, 2024 (T3) among students in two rural boarding junior high schools in Hebei Province, and a sample of 601 students who completed all the surveys was finally obtained. Students completed questionnaires, including the Delaware Bullying Victimization Scale Student (DBVS-S), the University of California at Los Angeles Loneliness Scale (UCLA), the Ruminative Responses Scale, and the Pittsburgh Sleep Quality Index (PSQI). Group differences were examined by using t-test or ANOVA, correlations between variables were analyzed by using Pearson correlation coefficients, and a serial mediation structural equation model was constructed, with mediation effects tested via the Bootstrap method. Results: Female students scored higher on sleep quality than male students (7.47±2.70, 6.47 ±2.46, t =4.74, P <0.01). Pearson correlation analysis indicated that bullying victimization was positively correlated with loneliness, rumination, and sleep quality; loneliness was positively correlated with rumination and sleep quality; and rumination was positively correlated with sleep quality ( r =0.26, 0.33, 0.23; 0.39, 0.38; 0.54, all P <0.01). Mediation analysis showed that T2 loneliness had an independent mediating effect of 0.70 (95% CI =0.36-1.35) between T1 bullying victimization and T3 sleep quality, T2 rumination had an independent mediating effect of 1.34 (95% CI =0.71-2.45), and the serial mediation effect of T2 loneliness and T2 rumination was 0.64 (95% CI =0.37-1.13), accounting for 22.11% of the total effect (all P <0.01). Conclusions Bullying victimization adversely affects sleep quality among rural boarding junior high school students through a longitudinal chain mediating pathway involving loneliness and rumination. Psychological interventions should be strengthened for students who experience bullying to alleviate their loneliness and reduce rumination, thereby improving sleep quality.

Based on the theory of blood collateral， postoperative recurrence and metastasis of bladder cancer are considered to arise primarily from the binding of stasis and toxin， which accumulate and hide within the blood collaterals. Accordingly， treatment should focus on clearing and resolving the deeply concealed stasis toxin retained in the blood collaterals. The paired use of insect and vine medicinals may exert synergistic effects by simultaneously searching out and eliminating pathogenic factors， guiding the action of herbs to the channels， and unblocking the collaterals. Drawing on clinical practice， the stasis-toxin pathogenesis of postoperative recurrence and metastasis of bladder cancer can be divided into four stages including stagnation and astringent of collateral qi， formation of fixed stasis nests， transformation of persistent stasis into toxin， and deficiency of healthy qi with lingering toxin. Accordingly， four herb pairs are proposed for each stage based on conventional treatment， which are Dilong （Pheretima）-Daxueteng （Caulis Sargentodoxae）， Shuizhi （Hirudo）-Jixueteng （Caulis Spatholobi）， Wugong （Scolopendra）-Luoshiteng （Caulis Trachelospermi）， and Quanxie （Scorpio）-Qianjinteng （stephania）. Their potential modern pharmacological mechanisms are further discussed.

Objective: To analyze the clinical characteristics and management strategies of metronidazole-associated Stevens-Johnson syndrome (SJS), and to provide a clinical basis for medication safety during the oral perioperative period. Methods: The study adhered to the tenets of the Declaration of Helsinki and was approved by the Ethics Committee of Stomatological Hospital of Xi'an Jiaotong University. A retrospective analysis was conducted on a case of SJS following mandibular impacted tooth extraction, for which the patient had taken metronidazole. Additionally, a literature review was performed by searching databases including China National Knowledge Infrastructure, Wanfang, PubMed, and Web of Science to identify reported cases of metronidazole-associated SJS. Demographic features, medication regimens, latency periods, clinical signs, interventions, and prognosis were analyzed. Results: The patient presented with lip swelling, extensive erosion and ulceration of the oral mucosa, and severe odynophagia. Scattered erythematous macules and bullae were observed on the trunk and extremities, with a positive Nikolsky sign. Additionally, there was erosion of the perineal skin and glans penis mucosa, along with skin desquamation of the scrotum. The patient with SJS was managed in the hospital by discontinuing metronidazole and administering sequential treatments, including glucocorticoids, immunomodulators, intravenous immunoglobulin, and plasma exchange. The condition gradually improved; after a 14-month follow-up, only mild dry eye remained, with no significant scarring of the skin or mucosa. However, delayed recognition of early warning signs resulted in delayed drug withdrawal and intervention. The literature review identified 6 additional cases (3 males, 3 females) with a median age of 45.5 years (range 31-61). Indications for metronidazole included postoperative sepsis prophylaxis after duodenal ulcer perforation repair, digestive system infections, mucosal diseases, pelvic inflammatory disease, septic cellulitis with pneumonia, and gingivitis. Administration routes comprised oral (n = 2), intravenous (n = 3), and topical (n = 1). The latency period from drug administration to onset ranged from 0.25 to 7 days (median 4 days). All of the cases presented with oral mucosal erosion, and a positive Nikolsky’s sign was noted for five patients. Discontinuation of the causative drug was the cornerstone of treatment, supplemented by immunomodulatory therapy, symptomatic support, and multidisciplinary collaboration. Prognoses varied significantly: among the four patients treated with “drug withdrawal + glucocorticoids + supportive care,” three recovered fully and one improved; among the two patients receiving “drug withdrawal + simple supportive care,” one recovered and one died Conclusion Although metronidazole-associated SJS is rare, it progresses rapidly and frequently involves oral mucosal damage, typically occurring within one week of administering the medication. Early drug withdrawal and prompt initiation of glucocorticoid therapy can improve prognosis. During the oral perioperative period, strict adherence to metronidazole indications, enhanced monitoring for early warning symptoms, and the establishment of a multidisciplinary collaborative diagnosis and treatment model are essential to minimize the risk of severe adverse drug reactions.

ObjectiveTo investigate the impact of Buzhong Yiqitang combined with cisplatin on the proliferation of human lung adenocarcinoma （A549） cells through regulation of the pyruvate dehydrogenase kinase 1 （PDK1）/protein kinase B （Akt） signaling pathway and influence on glycolysis. MethodsTranscriptome sequencing （RNA-seq） was employed to compare the expression of glycolysis-related genes between A549 cells and cisplatin-resistant human lung adenocarcinoma cells （A549/DDP）. Small interfering RNA （siRNA） was employed to knock down PDK1， and the knockdown efficiency was verified by Western blot and Read-time PCR. The cell counting kit-8 （CCK-8） assay was used to assess the survival and viability of A549 cells under the following conditions： siRNA negative control+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）， siPDK1+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）， and siPDK1+cisplatin （128， 64， 32， 16， 8， 4， 0 μmol·L^-1）+Buzhong Yiqitang （10%）-containing serum. The 20% inhibitory concentration （IC₂₀） of the siRNA negative control+cisplatin group （7.832 μmol·L^-1） was calculated and used as the subsequent cisplatin concentration. Colony formation assay was performed to evaluate the proliferation of A549 cells. Lactate and adenosine triphosphate （ATP） assay kits were used to measure lactate and ATP production. The mitochondrial membrane potential was detected with the fluorescent probe JC-1. Western blotting was conducted to examine the expression levels of PDK1， phosphorylated （p）-Akt， Akt， pyruvate kinase M2 （PKM2）， glucose transporter 1 （GLUT1）， pyruvate dehydrogenase （PDH）， and lactate dehydrogenase A （LDHA）. Confocal immunofluorescence was employed to detect PDK1 and p-Akt. ResultsRNA-seq results identified PDK1 as a highly expressed differential gene in glycolysis metabolism between A549 cells and A549/DDP cells， and it was highly expressed in tumor cells. Gene Set Enrichment Analysis （GSEA） revealed upregulated and downregulated genes in glycolysis and gluconeogenesis pathways. Western blot and RT-qPCR confirmed that PDK1-si-2 had the highest transfection efficiency， with a PDK1 knockdown rate exceeding 60%. CCK-8 assay determined the half-maximal inhibitory concentration （IC₅₀） values for each group as （30.698±5.348）， （16.372±3.562）， （13.237±1.573） μmol·L^-1， while the IC₂₀ of cisplatin in siRNA negative control-transfected A549 cells was （7.832±0.672） μmol·L^-1. Compared with the siRNA negative control group， the siRNA negative control+cisplatin group showed decreased colony formation rate， reduced lactate production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 fluorescence intensity （P<0.05）. The siPDK1 group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， PKM2， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1 group， the siPDK1+Buzhong Yiqitang group showed decreased colony formation rate， reduced lactate production， downregulated protein levels of PKM2， GLUT1， and PDK1， and reduced PDK1 fluorescence intensity （P<0.05）. The siPDK1+cisplatin group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of PKM2， GLUT1， PDK1， and PDH， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. The siPDK1+cisplatin+Buzhong Yiqitang group demonstrated decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， PKM2， GLUT1， PDK1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1+Buzhong Yiqitang group， the siPDK1+cisplatin group showed decreased colony formation rate， downregulated protein levels of p-Akt and PDH， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. The siPDK1+cisplatin+Buzhong Yiqitang group exhibited decreased colony formation rate， reduced lactate production， increased ATP production， lowered mitochondrial membrane potential， downregulated protein levels of p-Akt， GLUT1， PDH， and LDHA， and reduced PDK1 and p-Akt fluorescence intensity （P<0.05）. Compared with the siPDK1+cisplatin group， the siPDK1+cisplatin+Buzhong Yiqitang group showed decreased colony formation rate， increased ATP production， and downregulated protein levels of p-Akt， PKM2， and LDHA （P<0.05）. ConclusionBuzhong Yiqitang combined with cisplatin can suppress lung adenocarcinoma cell proliferation by modulating glycolysis through the PDK1/Akt signaling pathway.

Perimenopause raises the risk and incidence of depression, whereas the underlying molecular mechanism remains unclear. Disturbed glucose regulation has been widely documented in depressive disorders, which renders the brain susceptible to various stresses such as estrogen depletion. However, whether and how glucose dysfunction regulates depression-like behaviors and neuronal damage in perimenopausal transition remains unexplored. Here, a prominent depressive phenotype was found in perimenopausal mice induced by the ovarian toxin 4-vinylcyclohexene diepoxide (VCD). The VCD depression susceptible group (VCD^SS) and the VCD depression resilient group (VCD^RES) were determined using a ROC-based behavioral screening approach. We found that the hippocampus, a crucial region linked to depression, had hyperglycemia and mitochondrial abnormalities. Interestingly, oral administration of the SGLT2 inhibitor empagliflozin (EMPA) and intrahippocampal glucose infusion suggest a close relationship between hyperglycemia in the hippocampus and the susceptibility to depression. We verified that cytochrome c oxidase 7c (COX7C) downregulation is a potential cause of the high glucose-induced neuronal injury using proteomic screening and biochemical validations. High glucose causes COX7C to be ubiquitinated in a S-phase kinase associated protein 1 (SKP1)-dependent manner. According to these results, SKP1/COX7C represents a unique therapeutic target and a novel molecular route for treating perimenopausal depression.

Objective This study aimed to analyze the characteristics and clinical significance of peripheral lymphocytic subsets and cytokine levels, including interleukin 1β(IL-1β), IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12P70, IL-17A, tumor necrosis factor α(TNF-α), interferon γ(IFN-γ) and IFN-α, in patients with primary biliary cholangitis (PBC), to provide some novel insights into the pathogenesis of PBC. Methods We retrospectively collected clinical features and laboratory data from hospitalized patients who were primarily diagnosed with PBC and from healthy physical examinees at the Third People's Hospital of Changzhou between January 1, 2023, and June 30, 2024. Results A total of 152 PBC patients and 96 healthy controls who met the inclusion and exclusion criteria were enrolled. Significant differences were observed in baseline characteristics and laboratory data between the two groups. After the propensity score matching (PSM) analysis, 61 PBC patients and 61 healthy controls were successfully matched, ensuring that the general characteristics (age and gender) of the two groups were balanced and comparable. Compared to the control group, the proportion of peripheral lymphocytes was significantly higher in the PBC group (31.9% vs. 17.8%), primarily due to an increase in CD4⁺ T cells (46.77% vs. 41.19%), while CD8⁺T cells were significantly decreased (19.73% vs. 22.07%). Notably, the proportions of CD4⁺ programmed cell death 1 (PD-1)⁺ T and CD8⁺PD-1⁺ T cells were elevated, with CD8⁺PD-1⁺ T cells showing a significant positive correlation with the severity of liver inflammation (r=0.41). Furthermore, the mitochondrial mass (MM) of CD4⁺ T cells was significantly increased in PBC patients, whereas no significant changes were observed in the MM of CD8⁺ T cells or the mitochondrial membrane potential (MMP) of CD3⁺ T cells. Additionally, the plasma levels of cytokines, such as IL-4, IL-8, IL-10 and IFN-α, were abnormally elevated. The plasma levels of IL-5 and IL-1β were negatively correlated with the stage of liver fibrosis in patients with PBC (r=-0.52). Conclusion The overactivation and proliferation of CD4⁺ T cells, along with the suppression of CD8⁺ T cell function and increased PD-1 expression leads to T cell exhaustion, indicating significant immunological alterations in PBC patients. These changes are closely associated with the disease progression. Additionally, cytokines are likely involved in the immune regulation process of PBC and may influence the pathogenic mechanisms of the disease. Regular monitoring of lymphocyte subsets and cytokine levels can help assess the immune status and disease activity in patients with PBC, thereby guiding the individualized treatment strategies.
Humans ; Male ; Female ; Middle Aged ; Liver Cirrhosis, Biliary/blood* ; Retrospective Studies ; T-Lymphocyte Subsets/immunology* ; Aged ; Cytokines/blood* ; Adult ; CD8-Positive T-Lymphocytes/immunology*

This study investigated the role of the nuclear factor of activated T cells c3 (NFATc3) in vascular smooth muscle cells (VSMCs) during aortic aneurysm and dissection (AAD) progression and the underlying molecular mechanisms. Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD. VSMC-NFATc3 deletion reduced thoracic AAD (TAAD) and abdominal aortic aneurysm (AAA) progression in mice, contrary to VSMC-NFATc3 overexpression. VSMC-NFATc3 deletion reduced extracellular matrix (ECM) degradation and maintained the VSMC contractile phenotype. Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9 (MMP9) and MMP2, promoting ECM degradation and AAD development. NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2 (eEF2) and inhibiting its phosphorylation in the VSMC cytoplasm. Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice. Cabamiquine-targets eEF2 and inhibits protein synthesis-inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice. VSMC-NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development, making it a novel potential therapeutic target for preventing and treating AAD.

Objective:To analyze the current status of global clinical trial for botulinum toxin (BTX) drugs, and to provide a reference for BTX drug research priorities and trends.Methods:All registered BTX drug-related clinical trials from the inception of the platforms until December 2024 were retrieved from the International Clinical Trials Registry Platform (ICTRP) and the Center for Drug Evaluation (CDE) Clinical Trial Registration and Information Disclosure Platform of the National Medical Products Administration of China (referred to as the CDE Platform). The data were statistically analyzed using Excel 2021. The analysis was conducted from aspects such as registration volume and annual trend, distribution of conducting countries, drug type, study type and recruitment situation, and indications.Results:A total of 2 053 clinical trials related to BTX were included. The total number of registered trials showed an increasing trend year by year. The country with the largest number of clinical trials was the United States, with 571 trials, while China ranked third with 190 trials. Among the drug formulations, the injection form accounted for the highest proportion (99.02%, 2 033/2 053). A few new formulations included topical ointments, nasal sprays, and eye drops. In terms of drug serotypes and sources, the wild-type BTX-A projects accounted for a relatively high proportion (97.86%, 2 009/2 053), and also included a few wild-type BTX-B, wild-type BTX-E, recombinant BTX-AB, and recombinant BTX-A projects. In terms of research types, 90.4% (1 856/2 053) were intervention studies, 87.5% (1 796/2 053) of the clinical trials did not restrict gender, 90.7% (1 862/2 053) of the clinical trials recruited subjects covering adults, and 9.3% (191/2 053) of the clinical trials only recruited minors. The research indications covered various disciplines, such as skin beauty, neurology, urology, orthopedics, and ophthalmology. In recent years, the types of diseases registered for clinical trials had expanded significantly on the basis of the originally approved indications.Conclusion:Innovative directions in BTX drug research, including BTX drugs of different serotypes, recombinant BTX based on recombinant gene technology, BTX formulations for non-injectable delivery, and innovative areas of clinical application, are driving its continued clinical research.

OBJECTIVE To evaluate the effectiveness and safety of warfarin anticoagulation therapy guided by gene test in patients undergoing left ventricular assist device （LVAD） implantation， and to analyze the influencing factors of warfarin anticoagulation efficacy. METHODS Patients who underwent LVAD implantation at the Heart and Vascular Center of Northern Jiangsu People’s Hospital from January 2023 to October 2024 and required warfarin anticoagulant therapy were selected as the study subjects. They were divided into genetic testing group （n＝51） and empirical treatment group （n＝17） based on whether they underwent CYP2C9 and VKORC1 gene test. The gene test group was given warfarin based on the predicted dose calculated by gene test， while the empirical treatment group was given warfarin by clinical doctors based on international normalized ratio （INR） experience， all patients were given warfarin once a day. Follow-up observation was conducted for 6 months to compare the effectiveness [time in therapeutic range（TTR）， the time required to reach INR for the first time， the incidence of embolic events， the incidence of INR＜1.5 events] and safety （the incidence of major and minor bleeding events，the incidence of INR＞3.5 events） of warfarin treatment between two groups of patients. According to whether the patient’s TTR was ≥60%， they were divided into TTR≥60% group （n＝20） and TTR＜60% group （n＝48）. Univariate and multivariate binary Logistic regression analysis were used to determine the factors affecting the anticoagulant effect of warfarin in patients. RESULTS The TTR of patients in the gene test group was significantly higher than that in the empirical treatment group （P＜0.05）. The incidence of INR＜1.5 events in the gene test group was significantly lower than in the empirical treatment group （P＜0.05）. The incidence of minor bleeding events and INR＞3.5 events in the gene test group were lower than in the empirical treatment group， but the difference was not statistically significant （P＞0.05）. The results of multivariate binary Logistic regression analysis showed that gene test was an independent protective factor for warfarin anticoagulant therapy [odds ratio （OR）＝10.842， 95% confidence interval （CI）： 1.211-27.037， P＝0.033]， and the combination of statins was an independent risk factor for warfarin anticoagulant therapy [OR＝0.196， 95%CI： 0.045-0.861， P＝0.031]. CONCLUSIONS Under the guidance of gene test， warfarin anticoagulation therapy for LVAD patients after implantation can improve TTR， shorten the anticoagulation target time， and has good safety； meanwhile， it should be noted that the combination of statins may enhance the anticoagulant effect of warfarin， thereby increasing the risk of bleeding in patients.

OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.