Objective:To study the role of cadherin 1 (CDH1) gene DNA methylation in autoimmune thyroiditis (AIT) in population from different water-iodine regions.Methods:From May to June 2019, the information of AIT cases and healthy individuals in Shandong Province were collected in three types of water-iodine regions: iodine-fortification (IF) region, iodine-adequate (IA) region and iodine-excess (IE) region. A case-control study design was applied to match 176 AIT cases (case group) with age, gender, body mass index, and place of residence in a 1 ∶ 1 ratio to 176 healthy individuals (control group). Fasting urine and whole blood samples were collected to test the contents of urinary iodine, thyroid function indicators [serum free triiodothyronine (FT 3), free thyroxine (FT 4), thyroid stimulating hormone (TSH)], and serum iodine. The DNA methylation levels of the target region of the CDH1 gene and its four CpG sites in whole blood were determined using methylation sequencing technology for target regions (MethylTarget TM). Results:The DNA methylation level of the target region of CDH1 gene in the case group was 0.832 ± 0.044, and that in the control group was 0.828 ± 0.049, there was no statistically significant difference between the two groups ( t = 0.76, P = 0.448). There was no statistically significant difference in DNA methylation levels of the four CpG sites in the target region of CDH1 gene between the case group and the control group ( P > 0.05). There was no statistically significant difference in the DNA methylation level of the CDH1 gene target region between the case group and the control group in IF, IA and IE regions ( P > 0.05). The detection results of DNA methylation levels at CpG sites in the target region of CDH1 gene in different water iodine regions showed that the DNA methylation level at site 83 in case group in IF region was higher than that in the control group ( t = 2.30, P = 0.023). However, there was no statistically significant difference in the DNA methylation levels of the four CpG sites between the case group and the control group in IA and IE regions ( P > 0.05). The DNA methylation level of CDH1 gene target region in AIT patients was not significantly correlated with urinary iodine, serum iodine, and serum FT 3, FT 4, and TSH contents ( P > 0.05), but was significantly negatively correlated with age ( r =-0.19, P = 0.014). Conclusions:The DNA methylation level at CpG site 83 of CDH1 gene in AIT patients in IF region is significantly higher than that in control population, indicating that DNA methylation at this locus may be involved in the occurrence and development of AIT after iodine fortification. The DNA methylation level of CDH1 gene is negatively correlated with age.

Objective To explore the differences in neural activity between patients with anorexia nervosa(AN)and healthy controls(HC),as well as the association between these differences and symptoms in AN patients,using activation likelihood estimation(ALE)meta-analysis.Methods The literature search covered the period from 2000 to March 2025.From a pool of 588 identified studies,4 studies focusing on the neural activity differences between AN patients and HC were selected for inclusion.These studies comprised 106 participants and 21 sets of coordinates.The ALE meta-analysis method was employed,and the GingerALE software was used to systematically analyze the reported brain region changes and their peak coordinates,aiming to investigate the differences in brain functional activity between AN patients and HC.Results Compared to the HC group,AN group showed significantly enhanced activation in the left parahippocampal gyrus/amygdala(ALE value=0.39×10-2),right parahippocampal gyrus/amygdala(ALE value=0.39×10-2)and suboccipital gyrus(ALE value=0.39×10-2),along with a significant reduction in activation in Brodmann area 17(ALE value=0.61×10-2)(P<0.01,FWE corrected).Conclusion Key brain regions in AN patients including the parahippocampal gyrus,inferior occipital gyrus,and amygdala demonstrate significant functional activation abnormalities.

Objective:To analyze the role of interleukin-33(IL-33)in the occurrence and development of glioma and its related mechanism by bioinformatics technology,and to validate it through histopathological experiments,and to discuss the possibility of IL-33 as an auxiliary marker for the diagnosis and treatment of brain glioma.Methods:The glioblastoma multiforme/lower grade glioma(GBMLGG)case data were downloaded from the UCSC XENA database,including data of 689 glioma samples,5 paracancerous samples,and 1 152 normal brain tissue samples;Mann-Whitney U test was used to analyze the difference in the expression of IL-33 mRNA between the GBMLGG samples and the normal brain tissues;according to the expression level of IL-33 in GBMLGG tissue,the tumor samples were divided into IL-33 low expression group and IL-33 high expression group;the Human Protein Atlas(HPA)was used to validate the difference in the protein expression of IL-33 in the GBMLGG samples;the R language DESeq2(v.1.36.0)package was used to screen the differentially expressed genes(DEGs)in the GBMLGG tumor case samples;Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were used to perform pathway analysis on the DEGs;Gene Set Enrichment Analysis(GSEA)was used to discuss the pathways significantly enriched by IL-33 in the GBMLGG tissues;GSVA package was used to analyze the immune infiltration in the GBMLGG samples;survival package and survminer package were used to analyze the effect of IL-33 expression level on the survival of the patients in different clinical subgroups of GBMLGG;univariate and multivariate Cox proportional hazards regression models were used to analyze the relationship between IL-33 expression and the clinicopathological characteristics of the GBMLGG patients;the GBMLGG and control tissue samples were collected;immunohistochemical staining was used to detect the expression levels of IL-33 and its receptor suppression of tumorigenicity 2(ST2)in the GBMLGG and normal brain tissue samples.Results:The expression levels of IL-33 mRNA and protein in the GBMLGG tissues were significantly increased compared with those in normal brain tissues;there were 634 DEGs in total between the IL-33 low and high expression groups,including 283 up-regulated DEGs and 351 down-regulated DEGs;the GO functional enrichment analysis and KEGG signaling pathway enrichment analysis results showed that the DEGs were associated with biological behaviors such as activation of the classical pathway of complement,immunoglobulin complex formation,and mediated immunoglobulin receptor binding;in the course of GBMLGG development,high expression of IL-33 could degrade valine,leucine,and isoleucine,induce limonene and pinene degradation,promote propanoate metabolism,and simultaneously activate the Leishmania infection pathway,NOD-like receptor signaling pathway,and allograft rejection pathway;the infiltration levels of dendritic cell(DC)and mast cell in the IL-33 high expression group were higher than those in IL-33 low expression group;the infiltration levels of eosinophil,helper T cell,and central memory T cell(Tcm)were lower than those in IL-33 low expression group;the expression level of IL-33 was positively correlated with the infiltration of γδT cell(Tgd),helperT cell,macrophage,eosinophil,Tcm,and effector memory T cell(Tem)(P＜0.05);it was negatively correlated with the infiltration levels of DC,natural killer cell(NK),CD8+T cell,and CD56bright NK cell(P＜0.05).There were no significant differences in the overall survival(OS),disease-specific survival(DSS),and disease-free interval(DFI)of the GBMLGG patients between IL-33 high expression group and IL-33 low expression group(P＞0.05);the clinical subgroup analysis results showed that the expression level of IL-33 in oligodendrocytoma tissues was lower than those in astrocytoma and oligoastrocytoma tissues,and the expression level of IL-33 in glioblastoma tissues was higher than that in oligodendroglioma tissues.World Health Organization(WHO)stage and age were risk factors affecting the prognosis of the GBMLGG patients,and IDH mutation and primary treatment effect were protective factors affecting the prognosis;The immunohistochemical staining results showed that compared with normal brain tissues,the expression levels of IL-33 and its receptor ST2 proteins in the malignant glioma tissues were significantly increased(P＜0.05),and their expression levels were positively correlated in both normal brain tissues and malignant glioma tissues(P＜0.05).Conclusion:The expression level of IL-33 in the glioma tissue is significantly increased,and high expression of IL-33 may be a potential factor for poor prognosis in the glioma patients.

Objective To explore the differences in neural activity between patients with anorexia nervosa(AN)and healthy controls(HC),as well as the association between these differences and symptoms in AN patients,using activation likelihood estimation(ALE)meta-analysis.Methods The literature search covered the period from 2000 to March 2025.From a pool of 588 identified studies,4 studies focusing on the neural activity differences between AN patients and HC were selected for inclusion.These studies comprised 106 participants and 21 sets of coordinates.The ALE meta-analysis method was employed,and the GingerALE software was used to systematically analyze the reported brain region changes and their peak coordinates,aiming to investigate the differences in brain functional activity between AN patients and HC.Results Compared to the HC group,AN group showed significantly enhanced activation in the left parahippocampal gyrus/amygdala(ALE value=0.39×10-2),right parahippocampal gyrus/amygdala(ALE value=0.39×10-2)and suboccipital gyrus(ALE value=0.39×10-2),along with a significant reduction in activation in Brodmann area 17(ALE value=0.61×10-2)(P<0.01,FWE corrected).Conclusion Key brain regions in AN patients including the parahippocampal gyrus,inferior occipital gyrus,and amygdala demonstrate significant functional activation abnormalities.

Objective:To study the role of cadherin 1 (CDH1) gene DNA methylation in autoimmune thyroiditis (AIT) in population from different water-iodine regions.Methods:From May to June 2019, the information of AIT cases and healthy individuals in Shandong Province were collected in three types of water-iodine regions: iodine-fortification (IF) region, iodine-adequate (IA) region and iodine-excess (IE) region. A case-control study design was applied to match 176 AIT cases (case group) with age, gender, body mass index, and place of residence in a 1 ∶ 1 ratio to 176 healthy individuals (control group). Fasting urine and whole blood samples were collected to test the contents of urinary iodine, thyroid function indicators [serum free triiodothyronine (FT 3), free thyroxine (FT 4), thyroid stimulating hormone (TSH)], and serum iodine. The DNA methylation levels of the target region of the CDH1 gene and its four CpG sites in whole blood were determined using methylation sequencing technology for target regions (MethylTarget TM). Results:The DNA methylation level of the target region of CDH1 gene in the case group was 0.832 ± 0.044, and that in the control group was 0.828 ± 0.049, there was no statistically significant difference between the two groups ( t = 0.76, P = 0.448). There was no statistically significant difference in DNA methylation levels of the four CpG sites in the target region of CDH1 gene between the case group and the control group ( P > 0.05). There was no statistically significant difference in the DNA methylation level of the CDH1 gene target region between the case group and the control group in IF, IA and IE regions ( P > 0.05). The detection results of DNA methylation levels at CpG sites in the target region of CDH1 gene in different water iodine regions showed that the DNA methylation level at site 83 in case group in IF region was higher than that in the control group ( t = 2.30, P = 0.023). However, there was no statistically significant difference in the DNA methylation levels of the four CpG sites between the case group and the control group in IA and IE regions ( P > 0.05). The DNA methylation level of CDH1 gene target region in AIT patients was not significantly correlated with urinary iodine, serum iodine, and serum FT 3, FT 4, and TSH contents ( P > 0.05), but was significantly negatively correlated with age ( r =-0.19, P = 0.014). Conclusions:The DNA methylation level at CpG site 83 of CDH1 gene in AIT patients in IF region is significantly higher than that in control population, indicating that DNA methylation at this locus may be involved in the occurrence and development of AIT after iodine fortification. The DNA methylation level of CDH1 gene is negatively correlated with age.

Objective:To investigate the characteristics of bacterial community in upper gastrointestinal tumors.Methods:The study population was patients with upper gastrointestinal tumors (esophageal cancer and gastric cancer). Gastroscopy was performed on the enrolled patients ( n=17), and the specimens were taken from the tumor sites. At the same time, non-tumor tissues more than 4 cm away from the tumor tissues were taken as the control. After total DNA was extracted and purified, high-throughput 16S DNA gene sequencing was used to detect the microbiota in tumor tissues and control tissues. Bioinformatics analysis was carried out and the differences between groups were compared. Results:16S DNA PCR showed that there was no significant difference in bacterial load between tumor tissues and control tissues. The α-diversity and β-diversity indexes showed that the community composition of the two groups was similar; the samples were discrete and the colony composition was different, but there was no significant difference between the two groups. The results of Venn diagram showed that there were more operational taxonomic units (OTUs) in non-tumor tissues than in tumor tissues (2 068 vs 1 358), indicating that the bacterial species in normal tissues were more abundant than those in tumor tissues. Compared with the control tissues, the percentages of Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium) in tumor tissues were relatively higher (the average percentage was more than twice that of the control). Further paired comparison of the top ten bacteria in the family and genus abundance of the two groups of samples showed that Pseudomonas decreased significantly in tumor tissues at the family ( P=0.041) and genus ( P=0.041) levels, while Prevotella was significantly enriched in tumor tissues at the family ( P=0.031) and genus ( P=0.007) levels. Conclusions:The bacterial community in the tumor microenvironment of the upper gastrointestinal tumor changed, and the species enriched in the tumor site were mainly oral common anaerobic bacteria, such as Prevotellaceae ( Prevotella), Lactobacaceae ( Lactobacillus) and Fusobacteriaceae ( Fusobacterium), especially Prevotellaceae ( Prevotella).

Objective:To observe the clinical characteristics of patients with familial vitreous amyloidosis (FVA) and the efficacy of vitrectomy (PPV) and the occurrence of complications.Methods:A retrospective clinical study. From June 2009 to March 2020, 32 eyes of 18 patients from 3 FVA families who were diagnosed and treated by PPV at Department of Ophthalmology of Jiaxing TCM Hospital were included in the study. Among them, there were 12 males with 22 eyes and 6 females with 10 eyes. The average age of onset was 42.28±3.25 years; the average duration of disease was 3.75±3.93 years. All the affected eyes underwent best corrected visual acuity (BCVA) and B-mode ultrasound examination. A logarithmic visual acuity chart was used in the BCVA examination, which was converted to the logarithmic minimum angle of resolution (logMAR) visual acuity when recorded. The average logMAR BCVA of the affected eye was 1.72±0.53; the intraocular pressure was less than 21 mm Hg (1 mm Hg=0.133 kPa). The vitreous body of the affected eye was obviously cloudy. All the affected eyes underwent standard three-channel PPV through the flat part of the ciliary body, and vitreous specimens were collected for pathological examination during the operation. Peripheral venous blood of probands from 3 families was collected, and the whole exome gene sequencing was performed. The follow-up time after surgery was ≥6 months. The patient's clinical characteristics, fundus lesions in PPV, changes in BCVA after surgery, and complications was observed. One-way analysis of variance or t test was performed for measurement data comparison; χ2 test was performed for count data comparison. Results:The vitreous body of the affected eye showed gray-white dense and thick flocculent changes, and the posterior capsule attached to the lens showed "foot disc-like" turbidity; later the lens was mainly cystic opacity. Pathological examination of the vitreous body showed positive staining of Congo red; under a polarized light microscope, it showed apple green dots and sheet-like birefringence. The genetic test results showed that there was a c.307G>C (p.Gly103Arg) missense mutation in the TTR gene of the proband in Family 2. Peripheral retinal hemorrhages in 4 eyes (12.5%, 4/32), retinal tears in 5 eyes (15.6%, 5/32), retinal degeneration in 4 eyes (12.5%, 4/32), retinal detachment were found in PPV 3 eyes (9.4%, 3/32). The vitreous body was filled with C 3F 8 and silicone oil respectively for 2, 1 eye. Six months after the operation, the logMAR BCVA of the affected eye was 0.39±0.32, which was significantly higher than that before the operation, and the difference was statistically significant ( t=15.131, P=0.000). After the operation, high intraocular pressure occurred in 2 eyes (6.3%, 2/32), secondary glaucoma in 1 eye (3.1%, 1/32), retinal detachment in 2 eyes (6.3%, 2/32), neovascular glaucoma (NVG) in 2 eyes (6.3%, 2/32), cataract in 10 eyes (31.3%, 10/32). Conclusion:The vitreous body of FVA eyes are gray-white dense, thick and flocculent, attached to the posterior lens capsule, showing "foot disc-like" turbidity; PPV treatment can effectively improve the BCVA of the FVA eyes; secondary glaucoma, secondary retinal detachment, NVG can occur after surgery.

[Abstract] Objective: To investigate the miR-423-5p expression in brain glioma tissues and cell lines, and its promotive effect on temozolomide (TMZ) chemoresistance by targeting PDCD5 (programmed cell death protein 5). Methods: Tumor tissues and matched peritumoral tissues were collected from 20 brain glioma patients who were surgically treated in the Department of Neurosurgery, Affiliated Hospital of Beihua University between January 2017 and December 2018. Glioblastoma cell lines (U251, U87, SHG-44) and human normal glial cell line HMC-3 were also used in the study. The relative expression of miR-423-5p and PDCD5 in brain glioma and peritumoral tissues and cell lines was detected by qPCR. The synthesized miR-423-5p mimics and miR-NC were respectively transfected into U251 and U87 cells; meanwhile, TMZ at different concentrations (50, 100, 150 and 200 μmol/L) were also used to treat the cells. Then, the chemoresistance of cells to TMZ were determined. MTT assay and colony formation assay were used to examine the proliferation of U251 and U87 cells, andWestern blotting was used to detect the expression of c-caspase 3, Bcl-2 and PDCD5 proteins in U251 and U87 cells. The targeting relationship between PDCD5 and miR-423-5p was validated through Dual luciferase reporter gene assay. Results: miR-423-5p was highly expressed in glioma tissues and glioma cell lines (all P<0.01). As compared with the miR-NC group, the proliferation and TMZ-chemoresistance of U251 and U87 cells in miR-423-5p mimics group significantly increased (all P<0.01). Dual luciferase reporter gene assay validated that miR-423-5p could bind with PDCD5 3' UTR to suppress the expression of PDCD5. Conclusion: High expression of miR-423-5p enhances the chemoresistance of glioma cells to TMZ, and miR-423-5p may serve as a potential therapeutic target in the treatment of brain glioma.

Objective: To examine the effects of Berberine(BBR)on inflammatory pathways related to endoplasmic reticulum stress(ERS)in the penumbra area following focal cerebral ischemia-reperfusion injury in type 2 diabetic rats. Methods: A total of 72 healthy male Sprague-Dawley rats were fed a high-fat, high-sugar diet and injected with streptozotocin to establish a type 2 diabetes mellitus model.The diabetic rats were randomly divided by digital lottery method into a Sham operation group(Sham group), a diabetic rat + BBR treatment group(BBR group), a diabetic middle cerebral artery occlusion(MCAO)model group(MCAO group), and a diabetic rats MCAO + BBR treatment group(MCAO + BBR group). Six rats were included in each group.The two treatment groups received prespecified doses of BBR through gastric perfusion at 48 h, 24 h before surgery, and 6h after surgery.The MCAO model was prepared by a suture occlusion method.The neurological deficit scores were performed, and the expression of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)was detected by enzyme-linked immunosorbent assay(ELISA). The mRNA expression of ERS marker protein GRP78 was detected by quantitative real time polymerase chain reaction(RT-qPCR), and the expression of ERS-related inflammatory pathway proteins[78 Glucoseregulated protein(GRP78)、Pancreatic endoplasmic reticulum Rinase(PERK)、nuclear factor-κB(NF-κB)]was detected by Western blot. Results: the cerebral ischemic penumbra area, after 2 h of ischemia and 24 h of reperfusion, the neurological deficit score in the MCAO group was higher than that in the MACO+ BBR group [(2.83±0.41)vs.(1.67±0.52), P<0.05], and the expression levels of pro-inflammatory cytokines(TNF-α and IL-1β)and ERS-related inflammatory pathway proteins(GRP78, PERK and NF-κB p65)were also significantly increased(all P<0.05). However, BBR treatment was able to alleviate the neurological dysfunction caused by cerebral ischemia-reperfusion in type 2 diabetic rats(P<0.05). Similarly, BBR treatment also reduced the expression levels of pro-inflammatory factors(TNF-α and IL-1β)and ERS-related inflammatory pathway proteins(GRP78, PERK and NF-κB p65)in the cerebral ischemic penumbra area(all P<0.05). Conclusions Through inhibiting ERS-related inflammatory pathways, BBR plays a neuroprotective role to alleviate cerebral ischemia-reperfusion injury in type 2 diabetic rats.

Objective To understand the etiology and clinical characteristics of candidemia, and the diagnostic value of(1, 3)-β-Dglucan. Methods This analysis included 31 patients with candidemia, 39 patients with Candida deep colonization, and 50 healthy subjects, who were enrolled from Wuyi Chinese Hospital Affiliated to Jinan University during the period from September 2015 to March 2018. Clinical data such as fungal culture, clinical features, and serum(1, 3)-β-D-glucan level were analyzed and compared between groups. Results Overall, 13 strains of Candida albicans and 18 strains of non-albicans Candida were isolated from the 31 patients with candidemia. The incidence of candidemia was associated with old age(>65 years), prior surgery(≤7 days), indwelling catheter(≥2 catheters and lasting ≥5 days), duration of mechanical ventilation(≥5 days)(P<0.05), but not associated with sex, underlying disease, prior use of anmicrobial and antifungal agents, immunosuppressant or glucocorticoid, or hypoproteinemia(P>0.05). Serum level of(1, 3)-β-D-glucan was significantly higher in candidemia patients than in the patients with candida deep colonization and healthy controls(P<0.05). The sensitivity and specificity of(1, 3)-β-D-glucan in diagnosing candidemia were 87.1％ and 96.0％, respectively. The level of(1, 3)-β-D-glucan was significantly higher in candidemia with bacterial infection than candidemia without bacterial infection(P<0.05). The level of of(1, 3)-β-D-glucan was also significantly higher in the candidemia caused by Candida albicans alone than that caused by non-albicans Candida alone(P<0.05). Conclusions Non-albicans Candida is more prevalent than Candida albicans in candidiasis, but Candida albicans is still the most important Candida species. The incidence of candidemia is asociated with advanced age(>65 years), prior surgery within 7 days, indwelling catheters(≥2 catheters and lasting ≥5 days), and mechanical ventilation(≥5 days). Serum(1, 3)-β-D-glucan level is valuable for early diagnosis of candidemia.