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To analyze the temporal changes in lipid levels in patients following acute pancreatitis(AP) and explore the factors associated with post-AP serum triglyceride(TG) level changes. Methods Patients diagnosed with AP at a Hospital in Yangzhou between January 2017 and December 2020 were included in this study. Clinical data were collected retrospectively, and lipid profile data from follow-up visits after discharge were tracked. The follow-up period extended until January 1, 2024. Kaplan-Meier(K-M) curves were used to stratify follow-up duration, while Spearman correlation analysis assessed the relationship between lipid levels and follow- up time. A linear regression model was used to investigate factors influencing TG level changes post-AP. Results A total of 141 patients with 306 follow-up visits were included. Spearman correlation analysis showed a mild increase in lipid levels over time post-discharge: TG(r=0.159, P=0.005), total cholesterol(TC)(r=0.231, P＜0.000 1), High-Density Lipoprotein Cholesterol(HDL-C)(r=0.181, P=0.002), and ApolipoproteinA1(ApoA1)(r=0.371, P < 0.000 1). In the univariate linear mixed model,male gender(β=0.160,P=0.007), Body Mass Index(BMI) (β=0.017, P=0.007), diabetes history(β=0.138, P=0.030), smoking history(β=0.166,P=0.004),and recurrent AP(β=0.119, P=0.029) were significantly associated with Lg(TG) levels (P<0.05). In the multivariate model, BMI(β=0.019, P=0.042), smoking history(β=0.155, P=0.049), and recurrent AP(β=0.148,P=0.032) remained significantly positively correlated with changes in Lg(TG) levels after AP, albeit with a low correlation strength (r < 0.200). Conclusion Lipid levels, including TG, TC, HDL-Cand ApoA1, tend to increase over time in AP patients after discharge, with this trend being more pronounced in those with hypertriglyceridemic acute pancreatitis. Post-AP TG levels are significantly influenced by BMI at the time of onset, smoking history and recurrent AP.

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To investigate the role mechanism of doublecortin (DCX) in glioma and screen DCX overexpression-related differentially expressed genes, so as to provide new targets for glioma targeted therapy.Methods Using rat glioma C6 cells as the target cells, LV-DCX-EGFP, LV-Cas9-Puro, and LV-Ctrl-EGFP lentiviruses were constructed. DCX overexpressing and control C6 cell lines were established through Cas9 dual- vector system transfection and screening. Transcriptome sequencing was used to obtain the differentially expressed gene profiles. iDEP and DESeq2 were applied to screen differentially expressed genes with the threshold of |log2FC|>2 and P<0.05. STRING and Cytoscape were utilized to construct protein-protein interaction networks and screen key genes. DAVID was employed for Gene Ontology (GO) enrichment analysis. BUSCA and UniProt were used to predict the subcellular localization of key genes. Receiver operating characteristic (ROC) curves were plotted to verify the diagnostic value of key genes. Enrichr was adopted to predict the regulatory networks of transcription factors and miRNAs. Real-time quantitative PCR (RT-qPCR) was performed to validate the expression of key genes. Results A total of 45 upregulated and 47 downregulated differentially expressed genes were screened, and 6 upregulated key genes including POU2F3 and 5 downregulated key genes including UBB were identified. GO enrichment analysis showed that upregulated genes were enriched in biological processes such as the positive regulation of lipid biosynthesis, whereas downregulated genes were linked to molecular functions including the regulation of alkaline phosphatase activity. Key genes were mainly distributed in the nucleus and cytoplasm. Except for RPS17, all key genes had an AUC value >0.7, indicating good diagnostic value. RT-qPCR validation showed that the mRNA expression levels of some upregulated genes (POU2F3,LPAR6, SREBF1) significantly increased (P<0.001), while the mRNA expression levels of some downregulated genes (UBB, ACTB, UBE2I) also significantly increased (P<0.001), which might be related to transcriptional and post-transcriptional regulation. Conclusion Differentially expressed genes and their regulatory networks related to DCX overexpression in glioma are successfully screened, providing a theoretical basis for revealing the role mechanism of DCX in glioma development and laying a foundation for the development of potential therapeutic strategies.

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To investigate the potential mechanisms of Xanthatin in inhibiting the proliferation of laryngeal squamous cell carcinoma (LSCC) cells by integrating network pharmacology and in vitro experiments. Methods The targets of Xanthatin were identified using databases such as PharmMapper, while disease-related targets for LSCC were obtained from databases such as DisGeNET. The overlapping targets between Xanthatin and LSCC were determined by intersecting these datasets. A protein-protein interaction (PPI) network was constructed based on the overlapping targets, and key targets were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of the overlapping targets were performed using R software. A "Xanthatin-target-pathway" network was visualized using Cytoscape 3.8.0 software. The preliminary validation of the aforementioned results was performed using molecular docking and transcriptomics. The effects of Xanthatin on the proliferation of TU177 cells were assessed using CCK-8 and colony formation assays. Additionally, Western blot analysis was employed to measure the expression levels of PI3K, p-PI3K, Akt, and p-Akt proteins. Results A total of 159 overlapping targets between Xanthatin and LSCC were identified, and seven key targets, including AKT1, were screened. GO enrichment analysis yielded 2,455 entries, and KEGG enrichment analysis identified 172 pathways, such as the PI3K-Akt signaling pathway. Xanthatin exhibited favorable binding activity with the core target proteins of LSCC in molecular docking experiments. The transcriptomics results showed high consistency with the predictions from network pharmacology. CCK-8 and colony formation assays demonstrated that Xanthatin at concentrations of 1, 2, and 4 μmol/L significantly inhibited the proliferation of TU177 cells in a dose-dependent manner. The expression levels of p-PI3K and p-Akt proteins decreased with increasing concentrations of Xanthatin. Conclusion Xanthatin may exert an inhibitory effect on the proliferation of laryngeal squamous cell carcinoma (LSCC) cells by modulating the PI3K-Akt signaling pathway.

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Migraine is a widespread neurovascular disorder, the pathogenesis of which is closely linked to the release of calcitonin gene-related peptide (CGRP), leading to a significant impairment in patients' quality of life.CGRP receptor antagonists exert their therapeutic effect by inhibiting the interaction between CGRP and its receptors, thereby preventing migraine attacks. Currently, several agents, including Rimegepant and Ubrogepant, have either received approval from the U.S. Food and Drug Administration or are in advanced stages of clinical trials. These drugs offer multiple advantages, such as the absence of vasoconstrictive effects, a rapid onset of action, and minimal interference with the immune system. Nevertheless, further investigation is necessary to assess their long-term safety, the potential emergence of drug resistance, and the development of individualized treatment protocols. Moreover, the integration of these novel therapies with existing treatment strategies remains a critical area for future research. This review aims to summarize recent national and international scientific advancements to establish a theoretical basis for the application of precision medicine in migraine management.

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To investigate the clinical characteristics and risk factors of Mycoplasma pneumoniae pneumonia (MPP) based on a dual classification integrating imaging features and clinical severity. Methods Medical records of 2054 pediatric patients with MPP were retrospectively analyzed. The cohort was stratified into severe consolidation (n=253), severe non-consolidation (n=118), non-severe consolidation (n=393), and non-severe non-consolidation groups (n=1 290) based on clinical and radiological findings. Inter group data and characteristics were compared and multiple regression analysis was conducted to construct a prediction model for severe consolidation group. Results Significant differences were observed among the groups in terms of age, duration of fever, length of hospital stay, presence of pulmonary rales, inflammatory markers (CRP, LDH), the use of hormones, and bronchoscopic treatment (all P < 0.05). Compared with the severe non-consolidation group, non-severe consolidation group, and non-severe non-consolidation group, children with severe consolidation pneumonia caused by MPP exhibited the longest duration of fever [8 (6, 11) days vs 6 (2, 9), 7 (6, 9) and 6 (3, 8) days, respectively] and the longest length of hospital stay [7 (5, 8) days vs 6 (5, 8), 6 (5, 8) and 6 (4, 7) days, respectively]. They also had the highest incidence of reduced breath sounds [34 cases (13.4%) vs 2 cases (1.7%), 29 cases (7.4%) and 13 cases (1.0%), respectively] and a substantially higher rate of coinfections, particularly viral infections [63 cases (24.9%) vs 23 cases (19.5%), 60 cases (15.3%) and 190 cases (14.7%), respectively]. Multivariate analysis indicated that the independent risk factors for severe MPP (SMPP) were age > 4.5 years, length of hospital stay > 6.5 days, reduced breath sounds, neutrophil-to-lymphocyte ratio (NLR) > 1.66, LDH > 370.5 u/L, CRP > 9.5 mg/L, and coinfection with viruses. Reduced breath sounds (OR = 5.58, 95% CI: 2.45 - 12.69) and coinfection with bacteria (OR = 3.11, 95% CI: 1.43 - 6.75) were identified as the most significant risk factors for pulmonary consolidation in non-severe MPP children. Additionally, reduced breath sounds, coinfection with viruses, LDH > 365.5 u/L, and CRP > 32.1 mg/L were risk factors for severe pneumonia in children with pulmonary consolidation. For non-consolidation MPP children, the presence of pulmonary dry rales (OR = 2.28, 95% CI: 1.46 - 3.56) was the primary independent risk factor for the development of severe pneumonia. Conclusion The chest imaging findings of MPP are associated with clinical severity, and the risk factor model constructed based on this imaging-clinical classification can assist in achieving precise hierarchical diagnosis and treatment in clinical practice.

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To investigate the effect of Toxoplasma gondii infection on copper metabolism in the kidneys of mice. Methods A total of 80 7-8-week-old C57BL/6 female mice were randomly divided into four groups of 20 mice in each group after one week of adaptation, including Control group, Cu group, TgCtwh6 group and Cu+TgCtwh6 group. Mice that were not infected and fed with normal diet and water were used as the Control group; Mice fed with 1 g/kg of copper chloride processing diet and 0.1% copper chloride water for 60 consecutive days were used as Cu group; Mice infected with 25-30 TgCtwh6 cysts (one of the predominant genotype Chinese 1 in China) fed with normal diet and water were used as the TgCtwh6 group; mice infected with 25-30 TgCtwh6 cysts and fed with a processed diet containing 1 g/kg of copper chloride and water with 0.1% copper chloride for 60 consecutive days were used as the Cu+TgCtwh6 group. ICP-MS was used to determine the changes in copper content in kidney tissues. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of mouse kidney tissue. The number of apoptotic cells was observed by PI staining. Western blotting was used to detect the protein expression levels of glutathione peroxidase 4 (GPX4) and superoxide dismutase (SOD1, SOD2).RT-qPCR was used to detect the mRNA expression of cuproptosis-related genes. Results Pathological manifestations such as inflammatory cell infiltration in the Cu group and TgCtwh6 group were seen under the microscope, and the inflammatory infiltrating cells of the renal interstitial were reduced in the Cu+TgCtwh6 group, and the pathological manifestations of glomerular tubular structure were improved. The number of apoptotic cells in the Cu+TgCtwh6 group (88.36±19) was lower than that in the Cu group (119.0±20). Compared with the Cu+TgCtwh6 group, the expression of SOD1 protein was down-regulated, and the difference was statistically significant (P<0.05). TgCtwh6 infection could restore the down-regulation of renal GLS expression and the up-regulation of ATP7B expression caused by copper overload. Conclusion Toxoplasma gondii infection can interfere with the copper metabolism pathway in the kidney of mice, improve the kidney damage caused by copper overload, and provide new clues for the treatment of copper overload disease.

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To investigate the characteristics of gray matter structure and clinical symptoms in patients with Alzheimer's disease (AD) comorbid with apathy (AD with apathy, AD-A). Methods The study included 30 patients with AD-A, 30 Alzheimer's disease patients without apathy (AD without apathy, AD-NA), and 30 healthy controls (HCs) matched in gender, age, and years of education. All participants underwent a comprehensive neuropsychological assessment and magnetic resonance imaging (MRI) scans. Voxel-based morphometry (VBM) was used to analyze changes in gray matter volume among the three groups. Additionally, the correlation between the identified abnormal brain regions and apathy scale scores was analyzed. Results There were no statistically significant differences among the three groups in terms of age, gender, years of education, or total intracranial volume. Compared with the HCs group, both the AD-A and AD-NA groups showed significantly lower scores in cognitive function (P<0.001). The AD-A group exhibited significantly higher apathy scale scores compared with the AD-NA group (P<0.001). Compared with the AD-NA group, the AD-A group showed reduced gray matter volume in the bilateral caudate nucleus, left orbitofrontal cortex, lingual gyrus, inferior frontal gyrus, superior frontal gyrus, entorhinal cortex, right middle frontal gyrus and posterior cingulate cortex (FWE-corrected, P<0.05 for all). Compared with the HCs group, the AD-A group exhibited reduced gray matter volume in the bilateral middle temporal gyrus, left fusiform gyrus, calcarine sulcus, postcentral gyrus, right inferior frontal gyrus and supramarginal gyrus (FWE-corrected, P<0.05 for all). Compared with the HCs group, the AD-NA group showed reduced gray matter volume in the left precuneus, inferior temporal gyrus, and right inferior temporal gyrus (FWE-corrected, P<0.05 for all). In the AD-A group, changes in the gray matter volume of the left caudate nucleus (r= -0.557, P=0.002) and right middle frontal gyrus (r=-0.620, P=0.001) were negatively correlated with the apathy evaluation scale (AES) scores. Conclusion Patients in the AD-A group exhibited significant atrophy in the frontal-temporal-basal ganglia circuit, and the degree of gray matter atrophy was correlated with the severity of apathy.

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To explore the effects of transmembrane protein 16B(TMEM16B) on brain edema and blood-brain barrier after cerebral ischemia-reperfusion injury in rats. Methods TMEM16B overexpression and knockdown was performed by adeno-associated virus (AAV), and then adult male Sprague-Dawley rats were subjected to middle cerebral artery occlusion (MCAO). Rats were randomly divided into Sham group, MCAO group, AAV no-load group, TMEM16B overexpression group and TMEM16B knockdown group. Modified neurological severity scores, adhesive removal test and cylinder test were used to evaluate neurologic function.The ultrastructure of ischemic brain tissue was observed by transmission electron microscope. Brain water content was reflected by dry wet weight ratio of brain tissue. The expressions ofTMEM16B, aquaporin4(AQP4), Claudin5 and zonula occludens-1 (ZO-1) were investigated by immunofluorescence and Western blot. Results Compared with the AAV no-load group, the sensory and motor functions of rats in TMEM16B overexpression group were significantly impaired. Mitochondria were swollen; mitochondrial cristae and tight junctions disappeared. The brain water content was higher in overexpression group. The expression ofTMEM16B and AQP4 increased while the expression of Claudin5 and ZO-1 decreased. Compared with the AAV no-load group, the rats in TMEM16B knockdown group showed some recovery in motor function. The mitochondrial cristae and structure were clear, and the basement membrane was partially blurred. The brain water content was lower in knockdown group. The protein levels ofTMEM16B and AQP4 were lower while the levels ofClaudin5 and ZO-1 were higher than that in AAV no-load group. Conclusion An increase in TMEM16B aggravates brain edema and blood-brain barrier damage in rats after cerebral ischemia-reperfusion injury, while a decrease in TMEM16B expression alleviates brain edema and protects the blood-brain barrier.

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To explore the association between mammalian sterile 20-like kinase 2 (MST2) gene polymorphism and haplotype and the risk of colorectal cancer, rectal cancer, and colon cancer in the Han population in Baotou area by case-control association study. Methods A total of 390 patients with colorectal cancer diagnosed by pathology and 413 normal physical examination pop- ulation were collected, and 2 mL of peripheral blood was taken for subsequent gene genotyping. Single nucleotide polymorphisms (SNPs) of MST2 gene were screened according to the genetic polymorphism data of Chinese Han population provided by the NCBI-Hapmap database. Gene genotyping was performed by Taqman method. Logistic regression was used to calculate the association between each SNP and the risk of colorectal cancer, colon cancer, and rectal cancer under codominant, dominant, overdominant, and recessive genetic models. Results Five SNPs ofMST2 gene were screened, namely rs11783149, rs10955176, rs7827435, rs4075986, rs3019295. Among them, SNP rs4075986 was associated with the risk of colorectal cancer. Compared with the rs4075986 GG+AA genotype, carrying the AG genotype [OR(95%CI)=2.473(1.844- 3.316) could increase the risk of colorectal cancer. Compared with the rs4075986 GG genotype, carrying the AG genotype [OR(95%CI)=2.475 (1.844-3.323) could increase the risk of colorectal cancer. SNP rs4075986 and rs3019295 were associated with the risk of rectal cancer. Compared with the rs4075986 GG+AA genotype, carrying the AG genotype [OR(95%CI)=3.411 (2.387-4.874)] could increase the risk of rectal cancer .Compared with the GG genotype, carrying the AG+AA genotype[OR(95% CI)=1.824 (1.262-2.638)] could increase the risk of rectal cancer. Compared with the rs3019295 GG+AA genotype, carrying the AG genotype [OR(95% CI)=0.706(0.501-0.996)] could reduce the risk of rectal cancer . SNP rs11783149 and rs4075986 were associated with the risk of colon cancer.Compared with the rs11783149 CC genotype, carrying the TT [OR(95%CI)=10.883(1.186-99.862)] and CT [OR(95%CI)=1.665(1.036-2.675)] genotype could increase the risk of colon cancer, respectively. Compared with the rs4075986 GG genotype, the AG+AA genotype [OR(95%CI)= 1.824(1.262-2.638)] could increase the risk of colon cancer. Conclusion MST2 gene SNP rs3019295 AG genotype may be protective factor for rectal cancer. SNP rs11783149 CT and TT genotypes maybe risk factors for colon cancer. SNP rs4075986 AG and AG+AA genotypes may be a common risk factors for colorectal cancer, rectal cancer and colon cancer.

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To explore the active components, key targets, and mechanism of action of turnip in alleviating pulmonary fibrosis (PF) based on network pharmacology and animal experiments. Methods The active components and targets of Brassica rapa L. were screened using the traditional Chinese medicine systems pharmacology database and analysis platform database, and PF-related targets were obtained from disease databases such as online mendelian inheritance of man (OMIM) and DrugBank. The intersection targets were used to construct a protein-protein interaction (PPI) network to identify core targets, followed by gene oncology (GO)/Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. In the animal experiments, a bleomycin-induced PF mouse model was established. Pathological changes in lung tissue were evaluated using HE and Masson staining. qRT-PCR was used to detect the mRNA expression of tumor necrosis factor-α (TNF-α), phosphatidylinositol 3-kinase (PI3K), and akstrain transforming 1 (AKT1), and immunofluorescence staining was used to measure the protein expression of TNF-α, PI3K, and AKT1. Results The 68 active components identified in Brassica rapa L. may regulate PI3K- Akt signaling pathway by acting on 89 potential targets such as TNF - α and AKT1. The results of animal experiments showed that polysaccharide of Brassica rapa L. (BRPs) could significantly reduce the degree of bleomycin induced pulmonary fibrosis in mice; HE and Masson staining of lung tissue showed that compared with the model group, the damage of alveolar structure, the infiltration of inflammatory cells and the deposition of collagen fibers in the BRPs treatment group were significantly reduced.Further mechanism studies showed that BRPs could significantly down-regulate the mRNA and protein expression levels of TNF-α, PI3K and AKT1 in lung tissue of pulmonary fibrosis mice. Conclusion Brassica rapa L. can synergistically alleviate pulmonary fibrosis through "multi-component, multi-target and multi-channel" approach;BRPs is one of the main active components, and plays an anti-fibrosis role by inhibiting TNF-α/PI3K Akt signaling pathway.

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To explore the role of tumour necrosis factor(TNF) in splenic infection of mice by Leishmania major. Methods To establish an infection model, promastigotes of Leishmania were injected intradermally into the right hind foot of mice. The thickness of the footpad and body weight were measured to monitor the infection. Histological changes in the spleen after infection were observed by HE staining. Changes in lymphocytes and monocytes in the spleen were detected by flow cytometry. The expression level of Arginase-1(Arg-1) and inducible nitric oxide synthase (iNOS) in the spleen was determined by indirect immunofluorescence. The effect of TNF-α on macrophage infection with Leishmania was evaluated in vitro. Results Compared to B6.WT mice, the spleens of B6.TNF-/- mice showed significant enlargement 42 days post-infection, with structural disruption. Various cells infiltrated and were dispersed throughout the entire spleen.Flow cytometry results indicated that after infection with Leishmania, there was no significant change in the proportions of T cells and B cells in the spleens of the mice, while CD11b monocytic cells significantly increased.Immunofluorescence results revealed that the M2 macrophage/monocyte marker Arg-1 was highly expressed in the spleens of B6.TNF-/- mice (P < 0.05). The expression of iNOS in the spleens of B6.WT mice was relatively strong (P < 0.05). In vitro studies found that the absence or inhibition of TNF-α significantly increased the infection of Leishmania by peritoneal macrophages (P < 0.01), while the addition of TNF-α markedly inhibited this infection (P < 0.01). Conclusion The splenic infection in B6.TNF-/- mice following subcutaneous inoculation of L. major in the hind footpad may be associated with the absence of TNF-α, which leads to M2-type differentiation of macrophages and reduced nitric oxide (NO) production.

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To investigate the role of murine double minute 2 (MDM2) in dihydroartemisinin’s (DHA)inhibition of lung adenocarcinoma cell proliferation and migration. Methods CCK8 assay was used to detect the inhibitory effect of gradient concentrations of DHA (0, 5, 10, 25, 50 and 100 μmol/L) and time gradients (0, 24, 48,and 72 h) on the proliferation of lung adenocarcinoma A549 and PC9 cells, and the half maximal inhibitory concentrate (IC50) were calculated respectively. Colony formation and scratch assays were used to detect the inhibitory effects of DHA on colony formation and migration of A549 and PC9 cells. Western blot was used to detect the inhibitory effects of DHA on MDM2 expression and epithelial-mesenchymal transition (EMT)-related proteins E-cadherin and N-cadherin. The promoting effects of MDM2 on proliferation, migration and EMT of lung adenocarcinoma cells were verified by small interfering RNA-mediated knockdown of MDM2 (si-MDM2). The reversal effects of MDM2 overexpression on DHA’s inhibition on the proliferation and migration of A549 and PC9 cells were observed. Results DHA inhibited the proliferation of A549 and PC9 cells in a dose- and time-dependent manner, with IC50 values of 30.57 and 78.61 μmol/L, respectively. Compared with the Control group, A549 and PC9 cells had significantly decreased colony formation (both P < 0.01) and migration (both P <0.01) upon treatment with DHA. Moreover, DHA significantly inhibited the protein expression levels of MDM2 and N-cadherin in A549 and PC9 cells, and upregulated the expression of E-cadherin protein (both P < 0.05).Compared with si-Control, si-MDM2 significantly inhibited the protein levels of MDM2 and N-cadherin in A549 and PC9 cells, and upregulated the expression of E-cadherin protein, and significantly inhibited the proliferation(both P < 0.01) and migration (both P < 0.01) of both cells. After overexpression of MDM2 in A549 and PC9 cells, the proliferation and migration ability were significantly enhanced (both P < 0.05), and the inhibitory effects of DHA were partially reversed by MDM2 overexpression (both P < 0.05). Conclusion DHA effectively inhibits the proliferation and migration of lung adenocarcinoma cells, and its mechanism is associated with the suppression of MDM2.

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To establish a mouse model of lung injury induced by self-antigens and explore its pathological mechanisms to provide a reliable animal model for studying the pathogenesis of rheumatoid arthritis-associated interstitial lung disease (RA-ILD). Method The mice were injected intradermal with antigenemulsion made of complete Freund's adjuvant (CFA) and lung tissue protein, and the emulsion prepared with incomplete Freund's adjuvant (IFA) was used to enhance immunity. HE staining was used to observe the histopathological changes in the mice. Masson staining was used to detect pulmonary fibrosis. The mRNA levels of rheumatoid factor (RF), Krebs von den Lungen-6 (KL-6) and surfactant protein D (SP-D) were detected by qPCR. The levels of ACPA, IgG1, IgG2a and IgG3 antibodies were detected by ELISA. The changes of inflammatory cells and α-smooth muscle actin (α-SMA)expression in lung tissue were detected by immunofluorescence staining. The protein expression levels of Collagen I and α-SMA were detected by Western blot. The changes of immune cells were detected by flow cytometry. Results HE staining showed that inflammatory cell infiltration increased and tissue structure changed significantly in lung tissue after the model was established by self-lung tissue antigen. Masson staining showed increased collagen deposition in lung tissue of model mice. qPCR tests revealed elevated mRNA levels of RF, KL-6 and SP-D.. ELISA tests revealed elevated levels of ACPA, IgG1 and IgG3 antibodies. Immunofluorescence results showed that monocytes and T cells increased, and α-SMA expression increased in the model group. Western blot results showed increased protein expressions of Collagen I and α-SMA. The flow cytometry results showed an increase in T cells and monocytes in the lung tissue. Conclusion The mouse model of lung injury induced by self-antigens is successfully established, and T cells and monocytes may be involved in the occurrence and progression of the disease.

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To explore the molecular mechanism of the therapeutic effect of total timosaponin on central precocious puberty (CPP) rats based on the RF-amide-related Peptide 3 (RFRP-3)/G protein-coupled receptor 147 (GPR147) signaling pathway. Methods Twenty-six day-old female SD rats were randomly assigned into a control group, a model group, an RFRP-3 up-regulated group (1.5 mg/kg), a total timosaponin group (200mg/kg), a total timosaponin+empty load group, and a total timosaponin+RFRP-3 knockdown group, with 12 rats in each group. A CPP rat model was prepared by subcutaneous injection of N-methyl-DL-aspartic acid (NMA) (40mg/kg). After modeling and administration, the opening time of the rat’s vulva and the first estrus interval daily were observed. Administration was stopped when the first estrus interval appeared in the model group. The day of vaginal opening and the time of the first estrus interval of rats were recorded in each group of rats. The wet masses of both ovaries and uterus were measured, and the ovarian and uterine indices were calculated; HE staining was used to observe the morphological changes of rat ovarian and uterine tissues, the number of corpus luteum in the ovaries and the thickness of the uterine wall were determined; ELISA method was applied to detect levels of serum luteinizing hormone (LH), follicle stimulating hormone (FSH), and estradiol (E2); RT-qPCR was used to detect the mRNA expression of hypothalamic gonadotropin-releasing hormone (GnRH), KISS-1, GPR54, RFRP-3, and GPR147. Results Compared with the control group, the day of vaginal opening and the time of the first estrus interval of rats in the model group were advanced (P<0.05), the ovarian index and uterine index, number of ovarian corpus luteum and uterine wall thickness, serum LH, FSH, E2 levels, and the GnRH, KISS-1, and GPR54 mRNA expression levels in the hypothalamus increased (P<0.05), while the RFRP-3 and GPR147 mRNA expression levels decreased (P<0.05). Compared with the model group, the day of vaginal opening and the time of the first estrus interval of rats in the RFRP-3 up-regulated group and the total timosaponin group were delayed (P<0.05), the ovarian index and uterine index, number of ovarian corpus luteum and uterine wall thickness, serum LH, FSH, E2 levels, and the GnRH, KISS-1, and GPR54 mRNA expression levels in the hypothalamus decreased (P<0.05), while the RFRP-3 and GPR147 mRNA expression levels increased (P<0.05). Knocking down RFRP-3 could prominently weaken the delaying effect of total timosaponin on the development of precocious puberty in CPP rats (P<0.05). Conclusion Total timosaponin may inhibit the release of GnRH and premature activation of the hypothalamic pituitary gonadal axis by activating the RFRP-3/GPR147 signaling pathway, thereby alleviating the degree of precocious puberty development in CPP model rats.

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To construct a prognostic model for gastric cancer using NETosis-related long non-coding RNAs (lncRNAs) and to investigate their expression and functional roles in gastric cancer progression. Methods Data from gastric cancer patients were obtained from the TCGA database, and 85 NETosis-related genes were identified from the GeneCards database. NETosis-associated lncRNAs were screened using Pearson correlation analysis. A lncRNA-based prognostic model was constructed and evaluated through survival analysis, ROC curves,C-index, and nomogram analysis. Differences in gene set enrichment between high- and low-risk groups were further explored. Additionally, RT-qPCR was performed to validate lncRNA expression in gastric cancer patients,and the CCLE database was utilized to investigate lncRNA expression across various tumor cell lines. Results A risk prognostic model for gastric cancer was constructed using 12 lncRNAs. Based on risk scores, samples were stratified into high- and low-risk groups across multiple datasets. Validation results demonstrated significantly worse survival outcomes in the high-risk group compared to the low-risk group, with excellent predictive performance of the model (AUC = 0.758, 95% CI: 0.688-0.828). Gene set enrichment analysis revealed that the high-risk group was enriched in gene sets strongly associated with inflammatory progression, whereas the low-risk group showed enrichment in gene sets related to normal DNA function. RT-qPCR confirmed differential expression of lncRNAs between tumor and adjacent normal tissues (P < 0.001). Furthermore, analysis of the CCLE database indicated substantial variations in lncRNA expression across different tumor cell lines. Conclusion The prognostic model constructed with 12 lncRNAs demonstrates potential for assessing prognosis and immune status in gastric cancer patients. Gene enrichment analysis provides insights into further mechanistic studies on NETosis. The differential expression of lncRNAs between cancerous and normal tissues, as well as across tumor cell lines, may inform novel subtype classification and therapeutic strategies.

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To explore the effect of tumor cell perineural invasion on the prognosis of cervical cancer and the early predictive factors of perineural invasion in patients with cervical cancer. Methods A retrospective analysis was conducted on the clinical, pathological, and survival data of 551 patients with cervical cancer. These patients were categorized into a survival group (n=477) and a death group (n=74). The baseline characteristics of the two groups were compared using independent samples t-tests, Wilcoxon rank-sum tests, and chi-square tests. Multivariate binary logistic stepwise regression analysis was employed to identify independent risk factors associated with mortality. In addition, univariate logistic regression analysis was performed to determine predictive factors for perineural invasion. A predictive model for perineural invasion in cervical cancer was subsequently developed based on the multivariate regression equation, and its predictive accuracy was assessed using the ROC curve. Results In the basic data of cervical cancer patients, the high level of perineural invasion, lymphatic metastasis and postoperative pathological stage in pathological data had an impact on the poor prognosis of patients (P<0.05), Lymphovascular space invasion, parametrial involvement, and tumor invasion depth ≥1/2 were identified as significant predictors of PNI. The predictive value was the best in the multivariate model (Area under the curve =0.80). Conclusion Perineural invasion is an independent risk factor for poor prognosis of cervical cancer patients, and the occurrence of perineural invasion can be effectively predicted by the constructed multivariate mode.

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To investigate the iodine nutrition status of children aged 8-10 in Guangming, Longhua and Yantian District of Shenzhen in 2023, and to explore the influencing factors of thyroid volume. To evaluate prevention strategies and to provide a scientific basis for the elimination of iodine deficiency disorders (IDD). Methods Urine and household salt samples were randomly collected from 580 non-boarding students aged 8-10 years for iodine content detection. Thyroid volume was measured using a fully digital ultrasound imaging system, and goiter prevalence was calculated. Results A total of 580 samples was tested. The median salt iodine concentration was 23.86 mg/kg, with 93.62% qualified iodized salt and 94.48% coverage rate. The median of urinary iodine was 265.00 μg/L, mainly distributed between 200 - <300 μg/L and ≥300 μg/L. The proportion of children with appropriate iodine was 20.86%, and the proportion of children with insufficient or excessive urinary iodine levels was 10.86% and 68.28% of the total surveyed population, respectively. The median thyroid volume was 3.27 mL, and the goiter rate was 1.72%. Multiple linear regression analysis showed that age was the risk factor for thyroid volume (β=0.328, P <0.05), while urinary iodine was the protective factor for thyroid volume (β= -4.134×10 -4, P <0.05). Conclusion The qualified iodized salt rate, median of urinary iodine, and goiter prevalence of 580 children aged 8-10 meet the elimination criteria for IDD. Age and urinary iodine are closely related to thyroid volume change. The urinary iodine level of children is generally high and requires serious attention.

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To determine the expression of melanoma-associated antigens D4 (MAGED4) and SIRT7 in human glioma, and to analyze the potential effects of MAGED4 and SIRT7 on glioma cell proliferation. Method The MAGED4 and SIRT7 expression levels and their correlation were compared by the China glioma genome atlas (CGGA), Human Protein Atlas (HPA), and UALCAN databases. Survival analysis, ROC curve analysis, and Cox regression analysis were used to predict the outcome of MAGED4 and SIRT 7 in glioma patients. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) signaling pathway enrichment analysis were used to explore the biological functions of MAGED4 and SIRT7 in glioma. Western blot experiment was used to investigate whether MAGED4 protein exerted its regulatory effects on the activity of the PI3K / AKT signaling pathway via SIRT7. The effect of MAGED4 on cell proliferation in glioma through SIRT7 was explored by CCK-8. Result The analysis results of CGGA, UALCAN, and HPA databases showed that the expression levels of MAGED4 and SIRT7 in glioma tissues were higher than those in normal brain tissue, and the expression were positively correlated. Results of survival, ROC, and Cox analysis showed that high expression of MAGED4 and SIRT7 mRNA were risk factors for poor prognosis in glioma. Results of KEGG enrichment analysis showed that MAGED4 and SIRT7 were associated with the PI3K/AKT signaling in glioma, and Western blot results showed that MAGED4 activated the PI3K/AKT signaling pathway by regulating SIRT7. The CCK-8 results showed that MAGED4 affected the proliferation of glioma cells through SIRT7. Conclusion MAGED4 and SIRT7 are highly expressed in glioma and associated with poor prognosis, and MAGED4 promotes glioma cell proliferation through activation of the PI3K/AKT signaling pathway by SIRT7.

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To investigate independent risk factors for lower extremity amputation (LEA) in hospitalized patients with diabetic foot ulcers (DFUs). Methods A multicenter retrospective analysis was conducted on the clinical data of 329 hospitalized patients with diabetic foot ulcers from four general hospitals across the nation. A multivariate logistic regression model was constructed, and prediction analysis was performed using R 4.2.1. The discriminative ability of the model was assessed using receiver operating characteristic (ROC) curves, while calibration accuracy and clinical applicability were evaluated via calibration curves and decision curve analysis. Results The study revealed that patients with higher education backgrounds showed lower disease severity (Wagner grade) (Z = -1.237, P < 0.05), while those with a history of coronary heart disease exhibited higher Wagner scores (Z = -2.084, P < 0.05). In the amputation prognosis analysis, prolonged duration of diabetes and elevated white blood cell count were positively correlated with amputation risk (both P < 0.01). Multivariable regression identified non-higher education, low hemoglobin levels, decreased total cholesterol, and abnormally elevated platelet counts as independent risk factors for high Wagner grades (≥grade 3) (all P < 0.05). The integrated predictive model incorporating these factors demonstrated strong discriminative performance, with an area under curve (AUC) of 0.880 (95% CI: 0.801 -0.960). The calibration curve slope approached the ideal value, and decision curve analysis confirmed the model’s clinical net benefit within a threshold probability range of 10% -65%. Conclusion Lower education level, poor baseline nutritional status, infection, hypercoagulability, and underlying vascular diseases collectively constitute key factors contributing to elevated amputation risk in DFU patients. The developed predictive model exhibits high accuracy and may assist clinicians in formulating individualized intervention strategies.

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To investigate the effect and mechanism of long non-coding RNA RMRP (LncRNA RMRP) on oxygen-glucose deprivation/reperfusion (OGD/R) -induced ferroptosis in mouse HL-1 cardiomyocytes by regulating miR-766-5p. Methods HL-1 cells were cultured in vitro, and OGD/R models were established. The expression levels of LncRNA RMRP in HL-1 cells at various reperfusion time points were subsequently quantified using qRT-PCR. The LncRNA RMRP small RNA interference fragment (si-RMRP) and its corresponding negative control (si-NC), as well as the miR-766-5p inhibitor and its respective negative control (inhibitor-NC), were transfected into HL-1 cells. Subsequently, the cells were subjected to OGD/R treatment. CCK-8 assay was employed to evaluate cell viability. Assay kits were employed to measure the levels of lactate dehydrogenase (LDH) in the cell supernatant, as well as the intracellular levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and ferrous ion (Fe2+). qRT-PCR analysis was conducted to assess the expression levels of LncRNA RMRP and miR-766-5p. Western blot analysis was conducted to assess the expression levels of proteins associated with ferroptosis including GPX4, SLC7A11, and FTH1. Dual-luciferase reporter assays were performed to investigate the sponge adsorption relationship between LncRNA RMRP and miR-766-5p. Results As reperfusion time extended, the expression level of LncRNA RMRP in cells progressively increased (P<0.01). Treatment with OGD/R significantly inhibited the viability of HL-1 cells, reduced the expression of miR-766-5p (P<0.01), elevated the levels of LDH in the supernatant, as well as MDA and Fe2+ levels within the cells, and decreased the activities of SOD and GSH in cells (P<0.01). Additionally, OGD/R treatment downregulated the protein expression levels of GPX4, SLC7A11, and FTH1(P<0.01). Silencing LncRNA RMRP reversed these effects by enhancing the viability of HL-1 cells, increasing miR-766-5p expression (P<0.01), reducing LDH in the supernatant, as well as MDA and Fe2+ levels within the cells, and promoting SOD and GSH activities in cells (P<0.01). Furthermore, silencing LncRNA RMRP upregulated the protein expression levels of GPX4, SLC7A11, and FTH1(P<0.01). The dual-luciferase reporter assay confirmed that LncRNA RMRP could regulate the expression of miR-766-5p through a sponge adsorption mechanism. Partial inhibition of miR-766-5p inhibitor expression could mitigate the improvement effect caused by LncRNA RMRP silencing on OGD/R-induced ferroptosis in HL-1 cells. Conclusion Silencing LncRNA RMRP inhibits OGD/R-induced ferroptosis in HL-1 cells, potentially through the sponge-mediated regulation of miR-766-5p expression.

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To study the effects and potential mechanisms of the aquaporin 1 (AQP1) inhibitor acetazolamide (AZ) on the proliferation, apoptosis, and inflammatory response of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). Methods TNF-α-induced RA-FLS was served as in vitro RA model. MTT assay, IF staining, and EdU incorporation assay were applied to study AZ's effects on RA-FLS proliferation.Hoechst staining, flow cytometry analysis of Annexin V-FITC/PI-stained cells, and mitochondrial membrane potential detection experiments were used to detect cell apoptosis. ELISA and quantitative real-time PCR methods were used to measure pro-inflammatory cytokine production. Cell autophagy was evaluated using IF staining and mCherry-GFP-LC3B puncta assay. Western blot was performed to detect the levels of autophagy, apoptosis, and proliferation-related proteins. Moreover, we examined the role of autophagy inhibition in AZ's effects on RA-FLS by co-treating with the autophagy activator rapamycin (RAPA) or the autophagy inhibitor 3-methyladenine (3-MA). Results AZ dose-dependently inhibited cell proliferation, promoted apoptosis, and reduced the production of pro-inflammatory cytokines in RA-FLS. Furthermore, AZ suppressed cytoprotective autophagy in these cells, as evidenced by decreased LC3B levels (P < 0.05), increased P62 expression (P < 0.05), and reduced autophagic flux (P < 0.01). Particularly, AZ's beneficial effects were reversed by RAPA-induced autophagy activation and enhanced by 3-MA-induced autophagy inhibition. Conclusion This study provides the first evidence that AZ hinders cytoprotective autophagy, thereby alleviating the hyperproliferation, apoptosis resistance, and aberrant inflammatory response of RA-FLS, revealing the core role of autophagy inhibition in AZ's anti-RA effects.

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To predict and screen potential biomarkers of systemic lupus eythematosus (SLE) based on machine learning algorithms and structural biology, and to reveal their mechanisms of action and to provide new targets for disease diagnosis and treatment. Methods Four machine learning algorithms, Random Forest (RF), eXtreme Gradient Boosting (XGBoost),Support Vector Machine (SVM), Least Absolute Shrinkage and Selection Operator (LASSO),were used to analyze the gene expression data of SLE patients in GEO (datasets: GSE121239 and GSE11907) to analyze the gene expression data of SLE patients and screen key markers.Peripheral blood single nucleated cells (PBMCs) from SLE patients were collected and RT-qPCR was used to detect differential gene expression levels. Subsequently, GSEA enrichment analysis was used to identify biomarker-related pathways. CIBERSORT immune infiltration analysis and protein interactions network were applied to calculate the sample immune cell infiltration abundance. Single-cell data were analyzed for gene expression specificity in immune cells.Interaction relationships in combination with AlphaFold3 (AF3) were predicted. Results Multiple algorithms were screened together to identify the unique marker gene HERC5, and expression analysis of multiple datasets showed that HERC5 was highly expressed in SLE compared to the normal group (P < 0.05), and RT-qPCR verified the same trend (P=0.006 2). Functional enrichment analysis identified the major pathway promoted by HERC5 in SLE as the IFN receptor signalling pathway (P < 0.05). Immune infiltration analysis showed that HERC5 was closely associated with immune cells (Neutrophils: r = 0.39, P < 0.05; Memory B cells: r = 0.33, P < 0.05;Activated dendritic cell：r = 0.52, P < 0.05). Most HERC5-related interacting proteins were associated with SLE, and potential transcription factors of HERC5 and its related genes were also significantly associated with immune responses. Conclusion The HERC5 gene is an important biomarker for SLE, which upregulates the IFN pathway to promote SLE progression and provides a new target for SLE diagnosis and treatment.

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To identify ferroptosis-related genes associated with gastric cancer prognosis and investigate their potential molecular functions. Methods Gene expression profiles and clinical information of gastric cancer tissues and adjacent normal tissues were obtained from TCGA database. Differential expression analysis of ferroptosis-related genes was performed using the "DESeq2" package in R software. Key genes were identified and a prognostic model for gastric cancer was constructed through Cox regression analysis based on the LASSO algorithm. Patients were stratified into high-risk and low-risk groups according to the median risk score. The accuracy of the model was evaluated using Kaplan-Meier survival analysis and ROC curve analysis. Immune cell infiltration in gastric cancer patients was assessed with the "CIBERSORT" package. The mRNA expression of differentially expressed genes (DEGs) with prognostic significance was examined in both gastric cancer and adjacent normal tissue samples. In vitro experiments were conducted to validate the impact of hydroxycarboxylic acid receptor 1 (HCAR1) on the malignant biological behavior of gastric cancer. Results Based on ferroptosis-related genes from the TCGA database, a novel prognostic model was constructed. It demonstrated robust predictive power for survival in both training and validation cohorts. RT-qPCR analysis of 8 pairs of gastric cancer and normal tissues revealed that the expression patterns of 6 prognostic DEGs in cancer tissues were consistent with those predicted by the model. In vitro experiments confirmed that downregulation of the key gene HCAR1 could inhibit the proliferation, invasion, and metastasis of gastric cancer cells. Conclusion The ferroptosis-related gene based prognostic model exhibits robust predictive capability, allowing for accurate determination of prognosis and survival in individuals with gastric cancer.

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Inhibitor of DNA binding 1 (ID1) is a crucial regulator of cell differentiation and plays a significant role in maintaining normal hematopoietic differentiation and development. Due to the lack of DNA-binding motif, ID1 functions as a dominant-negative inhibitor of basic helix-loop-helix factors to antagonize their abilities to bind to DNA and transcriptionally regulate target genes. Abnormal expression of ID1 is strongly associated with various hematologic disorders, including myeloid and lymphoblastic leukemia, multiple myeloma and myeloproliferative neoplasms. ID1 acts as a potential oncogene by participating in multiple signaling pathways that promote the malignant proliferation, invasion and therapy resistance in leukemic cells. Significant strides have yielded promising antileukemic effects of ID1 inhibitors, both alone and in combination with targeted therapies against oncogenic signaling pathways. Here, we review the relationship between ID1 expression and the initiation and progression of blood disorders, and summarize the clinical significance of ID1 as a novel therapeutic target and potential prognostic biomarker for hematologic malignancies.

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To analyze the bioinformatics of domestic Clostridium difficile toxin B (TcdB) and prepare it to provide data support for the development of effective vaccines. Methods Using bioinformatics software such as Snippy, Blast, Muscle, and the dist.alignment() and hclust() functions in R, 1, 355 strains of Clostridium difficile from NCBI GenBank in China were compared and analyzed, and TcdB were grouped. The maximum likelihood tree and phylogenetic tree were beautified and displayed using iTOL. An online bioinformatics analysis website was used to predict and analyze the spatial structure and antigenic epitopes of the two largest subgroups, TcdB1 and TcdB2. The antigen protein TcdB was expressed and purified by prokaryotic system. Results According to the genotype of toxin B, the 1 355 prevalent strains of Clostridium difficile in China could be roughly divided into 12 subtypes, among which TcdB1 and TcdB2 were the main subtypes, accounting for more than 93.94% of all isolated strains, and about 17.20% of the strains were nontoxigenic or lack tcdB. The antigen epitope prediction of TcdB1 and TcdB2 showed that their antigen epitope distributions were basically the same, and many of them were distributed outside the C-terminal combined repetitive oligopeptides domains. Conclusion A specialized typing system for C. difficile TcdB in China has been established, and its main subtypes have been predicted for antigenic epitopes. The screened TcdB has been expressed for recombinant preparation.

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To explore the role and molecular mechanism of parthenolide (PTL) in regulating tumor- associated macrophage (TAM) polarization to inhibit the malignant progression of gallbladder cancer cells. Methods M0 macrophages were co-cultured with human gallbladder cancer cell line GBC-SD and treated with PTL, and the cells were divided into control group, TAM group, TAM+PTL group. Flow cytometry was used to determine the levels of CD86 and CD206 in macrophages. RT-qPCR and Western blot were used to determine the expressions of CD163, interleukin-10 (IL-10) and arginase-1 (Arg-1). ELISA and Western blot were used to determine the secretion of CC chemokine ligand 2 (CCL2) in the supernatant of GBC-SD cells and the expression of CCL2 in the cells. GBC-SD cells were transfected with a CCL2 overexpression vector and divided into TAM, TAM+PTL, TAM+PTL+OE-NC, TAM+PTL+OE-CCL2 groups. Flow cytometry was used to assess CD86 and CD206 levels in macrophages, RT-qPCR and Western blot were used to assess CD163, IL-10 and Arg-1 expression. CCK-8 assay, plate-based colony formation assay and Transwell chamber assay were used to assess GBC-SD cell proliferation, migration and invasion, respectively. Results Compared with the TAM group, the TAM+PTL group showed increased CD86 expression and decreased CD206 expression in macrophages, the mRNA and protein expressions of CD163, IL-10 and Arg-1 were downregulated, the CCL2 content and protein expression in GBC- SD cells decreased, cell proliferation activity, colony formation, migration and invasion were all reduced (P<0.05). Compared with the TAM+PTL group, the TAM+PTL+OE-CCL2 group showed decreased CD86 expression and increased CD206 expression in macrophages, the mRNA and protein expressions of CD163, IL-10 and Arg-1 were upregulated. Cell proliferation activity was enhanced, and colony formation, migration, and invasion all increased (P<0.05). Conclusion Parthenolide inhibits the polarization of macrophages in TAMs to M2 type by downregulating CCL2 expression in gallbladder cancer cells, thereby preventing the proliferation, migration and invasion of gallbladder cancer cells.

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To investigate the effects of atorvastatin (ATV) on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs), periodontal ligament stem cells (PDLSCs), and dental pulp stem cells (DPSCs) in vitro, and to validate the regulatory effect of ATV on periodontal bone formation and tooth movement using a rat orthodontic tooth movement (OTM) model. Methods The effects of ATV on the proliferation and osteogenic/odontogenic differentiation of rat BMSCs, PDLSCs, and DPSCs were assessed in vitro. CCK-8 assay was used to detect the proliferation of the three types of cells. Alkaline phosphatase (ALP) staining and Alizarin Red staining were employed to evaluate osteogenic differentiation capacity. Western blot was used to detect the expression of osteogenesis-related proteins [Collagen type I (COL-I), Runt-related transcription factor 2 (Runx2), Bone morphogenetic protein-2 (BMP-2), Osteocalcin (OCN)] and the odontogenesis-related protein Dentin sialophosphoprotein (DSPP) in BMSCs, PDLSCs, and DPSCs. An OTM rat model was established, with rats randomly assigned to an ATV gavage group and a control group. The ATV gavage group received daily oral administration of ATV at a dose of 20 mg/kg, while the control group received an equal volume of solvent. Tooth movement distance was measured via Micro-CT on days 7, 14, and 21. Histomorphology of periodontal tissues was observed using Hematoxylin and Eosin (HE) staining and Masson staining. The gene and protein expression levels of osteogenic markers (BMP-2, Runx2, OCN) on the tension side of the first molar were detected by qRT-PCR and immunohistochemistry, respectively. Results ATV at concentrations of 10⁻⁶ mol/L and 10⁻⁷ mol/L significantly promoted the proliferation and osteogenic/odontogenic differentiation of BMSCs, PDLSCs, and DPSCs, manifested as enhanced ALP activity, increased mineralized nodule formation, and up-regulated expression of osteogenic/odontogenic proteins COL-I, Runx2, BMP-2, OCN, and DSPP (P<0.001). In the OTM model, compared with the control group, the ATV gavage group showed a significant reduction in tooth movement distance (P<0.05), enhanced osteogenic activity in periodontal tissues, and significantly increased gene (P<0.001) and protein (P<0.05) expression of BMP-2, Runx2, and OCN on the tension side of the first molar.

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To identify the independent risk factors for pyloric lymph node (PLN) metastasis in patients with upper gastric cancer (UGC) and to construct a nomogram prediction model applicable for UGC patients. Methods Clinical data of UGC patients treated at The First Affiliated Hospital of Anhui Medical University (n=721) and The First Affiliated Hospital of Anhui Medical University North District (n=102) between January 2020 and November 2023 were retrospectively collected. Patients were randomly divided into a training set (n=576) and a validation set (n=247) at a 7:3 ratio. Based on the training set, multivariate Logistic regression analysis was performed to identify independent risk factors for PLN metastasis, and a nomogram prediction model was constructed accordingly. The model’s discriminative ability and calibration were assessed using receiver operating characteristic (ROC) curves and calibration curves. Finally, external validation was conducted using the validation set to evaluate the model’s stability and generalizability. Results Multivariate Logistic regression analysis revealed that tumor size (OR=1.307, 95%CI: 1.038-1.646), T3 stage (OR=5.703, 95%CI: 1.271-25.581), T4 stage (OR=7.827,95%CI: 1.569-39.037), lymphovascular invasion (LVI) (OR=6.637,95%CI: 1.387- 31.574), differentiation extent (OR=3.160, 95%CI: 1.567-6.373), and fibrinogen degradation product (FDP) level (OR=4.565, 95%CI: 1.927-10.810) were independent risk factors for PLN metastasis in UGC patients. The nomogram model constructed based on these factors demonstrated areas under the ROC curve (AUC) of 0.815 (95%CI: 0.751-0.815) in the training set and 0.832 (95%CI: 0.731-0.933) in the validation set. Calibration curves indicated good agreement between predicted and observed outcomes. Conclusion This nomogram prediction model exhibits good predictive performance for assessing the risk of PLN metastasis in UGC patients.

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To investigate the effects of prostaglandin E2 (PGE2) microinjection into the median preoptic nucleus (MnPO) on core body temperature in female mice, and to clarify its underlying mechanism. Methods Microinjection cannula were implanted into the MnPO of female mice using stereotaxic surgery. Subsequently, a multi-channel temperature acquisition system was used to simultaneously monitor rectal and brown adipose tissue (BAT) temperatures before and after intra-MnPO injections of different reagents. To investigate the thermoregulatory effects of the microinjection of PGE2 into the MnPO, 12 female C57BL/6 mice were randomly divided into a saline group (n=6) and a PGE2 group (n=6), which were injected with 0.1 μL saline and PGE2 (2.8 mmol/L), respectively. To determine whether E-series prostaglandin receptor (EP)1, EP3, and EP4 receptors mediate the thermoregulatory effects of PGE2, 15 female C57BL/6 mice were randomly divided into 3 groups (n=5 per group). Each group received sequential microinjections into the MnPO of 0.1 μL of PGE2 (2.8 mmol/L), followed by a mixture of the respective EP antagonist (ant) (20 mmol/L)and PGE2 (2.8 mmol/L). Results Compared with baseline level, the rectal temperature (P<0.01) and BAT temperature (P<0.001) of female mice both increased significantly after microinjection of PGE2 into the MnPO. Compared with the saline group, the increases in rectal temperature (P<0.001) and BAT temperature (P<0.000 1) were significantly greater in the PGE2 group of mice. Furthermore, following the injection of PGE2 into MnPO, the increase in BAT temperature was found to be significantly greater than that in rectal temperature in mice (P<0.001). Compared to the administration of PGE2 alone, co-injection of an EP3 ant + PGE2 into the MnPO of mice resulted in a significantly smaller increase in both rectal temperature (P<0.001) and BAT temperature (P<0.001). In contrast, the increases in rectal and BAT temperatures following MnPO injection of either EP1 ant + PGE2 or EP4 ant + PGE2 were not statistically significant (P>0.05 for both). Conclusion Injection of PGE2 into the MnPO elevates BAT and core body temperature in female mice via the EP3 receptor.

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To provide evidence for strengthening tuberculosis control in Inner Mongolia by analyzing changes and influencing factors in public awareness of tuberculosis prevention and control information among residents between the early stage and 17 years after the implementation of the directly observed treatment, short-course (DOTS) strategy. Methods Based on the "National Public Knowledge, Belief and Behavior Questionnaire on Tuberculosis Prevention and Control" designed by the Chinese Center for Disease Control and Prevention, a questionnaire survey was conducted among residents in Inner Mongolia using a multi-stage stratified random sampling method. Chi-square test was used for univariate analysis, and binary Logistic regression was employed to explore the influencing factors of public awareness of tuberculosis prevention and control information. Results The overall awareness rate of core information on tuberculosis prevention and control among the public was 67.7% in 2006, and it decreased to 49.2% in 2023 (P<0.05). Multivariate Logistic regression analysis showed: Compared with the groups aged 15~29, illiterate and semi-illiterate, those with public medical care, and those with three or fewer family members, the awareness rate of the groups aged 60 and above (OR=0.689), those with primary and junior high school education (OR=0.856), and those with self-paid medical care (OR=0.468) was significantly lower in 2006. The awareness rate was relatively high in groups with more than 3 family members (OR=1.236) (P<0.05). Compared with the groups aged 15~29, illiterate and semi- illiterate, married, Han ethnicity, employed, and those receiving public medical care, the awareness rate was higher among the groups aged 30~59 (OR=1.976), 60 and above (OR=2.224), those with high school education and above (OR=2.801), and single (OR=2.244) in 2023. However, the awareness rates of ethnic minorities (OR=0.737), the unemployed (OR=0.557), and self-funded medical care (OR=0.497) groups were significantly lower (P<0.05). Conclusion Public awareness of TB prevention knowledge in Inner Mongolia remains suboptimal. Strengthening diversified health education campaigns, addressing social stigma, and improving healthcare access are critical to enhance regional TB control outcomes.

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To verify the association between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis by constructing an in vitro co-culture system of testis. Methods Testicular tissue blocks from 20 ~ 25-day-old male rats were placed in an in vitro culture system, and the culture medium was replaced every 2 to 3 days. PCR was used to verify the expression of marker genes of various spermatogenic cells. RNA interference technology was employed to verify the correlation between the Ddit3-Trib3-Akt signaling pathway and rat spermatogenesis. Results The co-culture system could be continuously cultured for more than 2.5 months in vitro. RT-PCR showed that specific marker genes of spermatogonia, spermatocyte and spermoblast were expressed. The RNA and protein expression ofTrib3 and Akt changed after the knocking down of Ddit3 and Trib3, respectively. It demonstrated the existence of Ddit3-Trib3-Akt signaling pathway in rat spermatogenesis. Conclusion The culture time of more than 2.5 months indicates that the culture system can temporarily maintain the proliferation and differentiation of stem cells, and simultaneously maintain and stabilize spermatogenesis in a simple system. The successful validation of the Ddit3-Trib3-Akt signaling pathway also confirms that this culture system can be used to study possible molecular mechanisms of spermatogenesis in vitro.

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To explore the relationship between negative life events and depression severity in adolescent patients with depressive disorder, as well as the chain mediating role of insomnia symptoms and quality of life. Methods From January 2021 to December 2022, 374 outpatient patients and hospitalized patients with adolescent depressive disorders were enrolled. The Adolescent Life Event Scale (ASLEC), the Insomnia Severity Index (ISI), the World Health Organization Quality of Life Questionnaire Short Form (WHOQOL-BREF), and the Center for Epidemiology Depression Scale (CES-D) were used to evaluate the negative life event situation,insomnia symptoms, quality of life level and depression severity of the subjects, respectively. In addition, the PROCESS 4.0 macroprogram was used to analyze the chain mediating effect of insomnia symptoms and quality of life between negative life events and depression severity in patients with adolescent depressive disorder. Results The results of correlation analysis showed that there was a significant correlation between negative life events and insomnia symptoms, quality of life, and depression severity (all P<0.05). In addition, the results of chain mediation showed that negative life events had a significant direct effect on depression severity, with an effect size of 0.12 (P<0.001). Insomnia symptoms and quality of life played a mediating role in he relationship between negative life events and depression severity in patients with adolescent depressive disorders, with indirect effect sizes of 0.061 (95%CI: 0.040-0.087) and 0.100 (95%CI:0.065-0.136), respectively. It could also play a chain mediation role, and the effect size was 0.043 (95%CI: 0.026-0.063). Conclusion Negative life events experienced by patients with adolescent depressive disorder not only directly affect the severity of depressive symptoms, but may also indirectly exacerbate depression through insomnia symptoms and quality of life.

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To investigate the differential expression of integrin alpha-6(ITGA6) in abdominal wall endometriosis (AWE) tissues and its molecular mechanisms in regulating AWE.Methods 36 AWE lesions were designated as the experimental group, while 36 cases of normal endometrial tissues served as the controls. Differential expression of ITGA6 between the two groups was assessed through immunohistochemical (IHC) staining. Human ITGA6 gene-specific interference sequences were designed, synthesized, and packaged into lentiviral vectors to establish the Ishikawa cell line with ITGA6-knockdown. Similarly, the ITGA6-overexpression cell line was constructed using the coding sequence (CDS) of the gene. Real-time PCR and Western blot were performed to detect changes in EMT-related markers and angiogenesis-related indicators. Cell invasion and migration capabilities were assessed by Cell Scratch and Transwell assays. Furthermore, Western blot was conducted to profile PI3K/AKT pathway dynamics. Results Ectopic endometrial tissues exhibited a marked increase in the number of ITGA6-positive cells and their expression intensity compared to eutopic endometrium (each P < 0.001). Compared with the NC group, the ITGA6-knockdown group showed significantly reduced expression of N- cadherin, VEGF, and TGF-bata (P < 0.001; P < 0.01; P < 0.01), while E-cadherin expression was markedly increased (P < 0.001). Concomitantly, the invasion and migration capacities of ITGA6- low expression were significantly impaired (P < 0.001 for both), accompanied by a marked reduction in AKT and phosphorylated AKT(p-AKT) levels (all P < 0.001). Conversely, overexpressing ITGA6 resulted in opposite effects. Conclusion ITGA6 modulates EMT and angiogenesis in Ishikawa cells via the PI3K/AKT signaling pathway, thereby enhancing cell invasion and migration capabilities, which contributes to the pathogenesis of abdominal wall endometriosis.

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To investigate the expression of TIPE3 in colorectal cancer (CRC) and its correlation with tumor immune infiltration and prognosis . Methods The expression of TIPE3 in tumor and adjacent normal tissues from 110 CRC patients was detected using real-time fluorescence quantitative enzyme chain reaction ( qPCR) , Western blot (Western blot ) , immunohistochemistry (IHC) , bioinformatics analysis , and single-cell RNA sequen-cing (scRNAseq) . The association between TIPE3 expression and clinicopathological factors , prognosis , and im- mune cell infiltration was analyzed . Results Transcriptomic and proteomic analyses revealed that among the TIPE family , TIPE (P < 0. 05) and TIPE3 (P < 0. 001) were significantly upregulated in CRC tissues compared to adja- cent normal tissues , with TIPE3 showing the most pronounced increase . IHC further confirmed that TIPE3 expres- sion was significantly higher in CRC tissues ( P < 0. 001) . Kaplan-Meier survival analysis indicated that high TIPE3 expression was associated with reduced survival and poorer prognosis . Immune infiltration analysis demon- strated that patients with high TIPE3 expression exhibited fewer CD8 + T cells and CD20 + B cells but more CD66b + neutrophils , suggesting a correlation between TIPE3 and unfavorable tumor prognosis . Additionally , scRNA-seq of CRC tissues yielded results consistent with the IHC findings . Conclusion TIPE3 expression in CRC is associated with tumor immune infiltration , and its upregulation predicts poor prognosis , indicating its potential as a therapeutic and preventive target in CRC .

abstract:
To investigate whether TYRO protein tyrosine kinase-binding protein (TYROBP) affects the progression of diabetic kidney disease (DKD) through the extracellular signal-regulated kinase ( ERK) pathway. Methods Key genes in DKD were identified through bioinformatics analysis . Immunohistochemical staining and quantitative real-time PCR (qPCR) were used to validate the expression levels of TYROBP in a DKD mouse model and high glucose-stimulated NRK-52E cells . NRK-52E cell models with stable TYROBP overexpression/knockdown and their corresponding empty vector (ev) /scrambled sequence (ss) controls were established via lentiviral trans- fection . Cells were treated with 5 . 5 mmol/L or 30. 0 mmol/L glucose for 72 hours to mimic normal glucose (NG) and high glucose ( HG) conditions , respectively. High glucose medium containing 3 . 5 μmol/L FR180204 was used for ERK inhibitor intervention . The experiment included seven groups : ev + NG , ev + HG , oe-TYROBP + HG , ss + NG , ss + HG , sh-TYROBP + HG , and sh-TYROBP + HG + ERK inhibitor. Western blot was used to de- tect the expression levels of phosphorylated ERK/total ERK (p-ERK/ERK) , apoptosis-related proteins B-cell lym- phoma-2 (Bcl-2) and Bcl-2-associated X protein ( Bax) , and epithelial-mesenchymal transition ( EMT)-related proteins E-cadherin and α-smooth muscle actin ( α-SMA) . Tetramethylrhodamine ethyl ester (TMRE) staining and Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) flow cytometry were performed to as- sess mitochondrial membrane potential and apoptosis levels . Results Bioinformatics analysis identified TYROBP as a key gene in DKD . In vivo and in vitro validation showed increased TYROBP mRNA levels in DKD models . The results from the HG model indicated that , compared to the ev + NG/ss + NG group , the ev + HG/ss + HG group demonstrated increased p-ERK/ERK expression , reduced mitochondrial membrane potential , elevated apoptosis , and enhanced EMT. In TYROBP-perturbed NRK-52E cells , compared to the ev + HG group , the oe-TYROBP + HG group showed decreased p-ERK/ERK expression (P < 0. 01) , increased mitochondrial membrane potential (P < 0. 05) , reduced apoptosis (P < 0. 001) , and attenuated EMT; whereas compared to the ss + HG group , the sh- TYROBP + HG group exhibited increased p-ERK/ERK expression ( P < 0. 001) , decreased mitochondrial mem- brane potential (P < 0. 01) , elevated apoptosis (P < 0. 001) , and enhanced EMT. Furthermore , compared to the sh-TYROBP + HG group , the sh-TYROBP + HG + ERK inhibitor group displayed reduced p-ERK/ERK expression (P < 0. 01) , increased mitochondrial membrane potential ( P < 0. 001) , decreased apoptosis ( P < 0. 001) , and suppressed EMT. Compared with the scrambled sequence control + high glucose group , the TYROBP knockdown + high glucose group showed elevated p-ERK/ERK expression ( P < 0. 001) , reduced mitochondrial membrane potential (P < 0. 01) , increased apoptosis level (P < 0. 001) , and enhanced EMT. Compared with the TYROBP knockdown + high glucose group , the TYROBP knockdown + high glucose + ERK inhibitor group demonstrated decreased p-ERK/ERK expression (P < 0. 01) , restored mitochondrial membrane potential (P < 0. 001) , reduced apoptosis level (P < 0. 001) , and suppressed EMT. Conclusion TYROBP may regulate the ERK signaling path- way to modulate apoptosis- and EMT-related proteins , thereby influencing mitochondrial membrane potential , apop- tosis , and EMT in renal tubular epithelial cells and contributing to DKD progression .