Primary biliary cholangitis (PBC) is a chronic cholestatic liver disease characterized by autoimmune-mediated destruction of intrahepatic bile ducts. Emerging evidence suggests that circular RNAs (circRNAs) play regulatory roles in autoimmune diseases, but their involvement in PBC remains unclear. This study focused on the hsa_circ_0000711 and its potential mechanism in PBC pathogenesis. The study included 46 PBC patients, 40 healthy controls, and 40 patients with other liver diseases. Human intrahepatic biliary epithelial cells (HiBEpic) were treated with 1 mM glycochenodeoxycholic acid (GCDCA) to establish a PBC cell model. Hsa_circ_0000711 was overexpressed or knocked down using plasmid transfection and siRNA, respectively. Expression levels were analysed by qPCR/Western blot, cell viability by CCK-8, and hsa_circ_0000711-miR-185-5p interaction by luciferase assay. Serum hsa_circ_0000711 levels were significantly higher in PBC patients compared to healthy controls and other liver disease groups (p < 0.0001). GCDCA-treated HiBEpic cells showed increased expression of the E2 subunit of the pyruvate dehydrogenase complex (PDC-E2) and nuclear factor of activated T cells 3 (NFATc3), along with decreased cell viability. Overexpression of hsa_circ_0000711 increased PDC-E2 and NFATc3 expression and worsened cellular injury, while its knockdown reversed these effects. The dual luciferase assay confirmed that hsa_circ_0000711 directly binds to miR-185-5p, suppressing its activity and thereby relieving the repression of NFATc3. Hsa_circ_0000711 promotes PBC progression by sponging miR-185-5p and upregulating NFATc3, leading to bile duct epithelial cell injury. These findings highlight its potential as a novel diagnostic biomarker and therapeutic target for PBC.

Minimally invasive testing is essential for early cancer detection, impacting patient survival rates significantly. Our study aimed to establish a pioneering cell-free immune-related miRNAs (cf-IRmiRNAs) signature for early cancer detection. We analyzed circulating miRNA profiles from 15,832 participants, including individuals with 13 types of cancer and control. The data was randomly divided into training, validation, and test sets (7:2:1), with an additional external test set of 684 participants. In the discovery phase, we identified 100 differentially expressed cf-IRmiRNAs between the malignant and non-malignant, retaining 39 using the least absolute shrinkage and selection operator (LASSO) method. Five machine learning algorithms were adopted to construct cf-IRmiRNAs signature, and the diagnostic classifies based on XGBoost algorithm showed the excellent performance for cancer detection in the validation set (AUC: 0.984, CI: 0.980–0.989), determined through 5-fold cross-validation and grid search. Further evaluation in the test and external test sets confirmed the reliability and efficacy of the classifier (AUC: 0.980 to 1.000). The classifier successfully detected early-stage cancers, particularly lung, prostate, and gastric cancers. It also distinguished between benign and malignant tumors. This study represents the largest and most comprehensive pan-cancer analysis on cf-IRmiRNAs, offering a promising non-invasive diagnostic biomarker for early cancer detection and potential impact on clinical practice.

Background Previous studies have shown that EDCs may activate nuclear transcription factor, such as activator protein-1 (AP-1), nuclear factor of activated Tcells (NF-AT) and nuclear factor kappa B (NF-κB) in the process of immune damage. At the same time, some experts believed that estrogen may play an important role in this process. As a typical representative of EDCs, nonylphenol (NP) has not been reported. The aim of this work was to explore the relationship between the immune inflammatory damage and the changes in estrogen expression in male rats during the chronic exposure to NP at environmental concentrations. Sixty SPF Sprague–Dawley rats were divided into five groups ( n = 12 per group): blank control group (corn oil), low-dose NP exposure group (0.4 mg/kg/d), medium-dose NP exposure group (4 mg/kg/d), high-dose NP exposure group (40 mg/kg/d), and estradiol control group (E 2 : 30 μg/kg/d). Results Compared with the control group, rat spleen organ coefficient, number of spleen nodules, relative area of lymph nodes and white pulp were relatively reduced in the L (NP, 0.4 mg/kg) and H (NP, 40 mg/kg) exposure dose groups ( P < 0.001). Lymphocytes were rich in cytoplasm, mitochondria were swollen, part of the cristae was reduced, and rough endoplasmic reticulum was expanded. The serum levels of IgG ( P < 0.001) and IgM ( P = 0.002) showed a downward trend. The percentage of Th cells (CD3 + CD4 + ) was significantly decreased ( P < 0.001), and the percentage of B lymphocytes shows an opposite trend ( P < 0.001). Giemsa staining showed that the number of neutrophils ( P < 0.001) was increased. The expressions of estrogen receptor ER-α and ER-β protein in the spleen increased significantly ( P < 0.001). The expressions of AP-1 protein and NF-AT protein in the spleen were increased, and the expression of NF-KB protein was decreased ( P < 0.001). The expressions of IL-4, ER-α and ER-β ( P < 0.001) levels in serum increased. The mRNA-seq bioinformatics detection showed the final differentially expressed immune-inflammatory-related genes between the control and H-NP groups as follow: down-regulated: TLR4, Gata3, IL12, up-regulated: TNF-a, IL10, INOS. The mRNA expressions of ER-α, ER-β, NF-KB, IL4, AP-1, TLR4, Gata3, and NF-AT were consistent with the results of mRNA-seq analysis. NP content was correlated with the expressions of ER-α, ER-β, IL4, AP-1, NF-AT, TLR4, NF-KB, as well as IL-12 proteins in the spleen tissue ([ r ] < 1, P < 0.05). Conclusions Chronic exposure to NP at environmental concentration could cause immune dysfunction, resulting in immunotoxicity and inflammatory effects, and lead to changes in the activity of transcription factors and differential immune inflammatory factors in rats. Graphical Abstract

Endocrine-disrupting chemicals (EDCs) might increase the risk of childhood diseases by disrupting hormone-mediated processes that are critical for growth and development during childhood, however, the association among the exposure level of EDCs such as Nonylphenol (NP), Bisphenol A (BPA), Dimethyl phthalate (DMP) in children and environmental risk factors, as well as hepatic function has not been elaborated. This study aimed to discuss this interesting relationship among NP, BPA, DMP concentrations in serum, environmental risk factors, hepatic function of 5- to 14-year-old children in industrial zone, residential zone and suburb in northern district of Guizhou Province, China. In Zunyi city, 1006 children participated in cross-sectional health assessments from July to August 2018, and their parents completed identical questionnaires on the environmental risk factors of EDCs exposure to mothers and children. Serum NP, BPA and DMP concentrations were measured by high performance liquid chromatography (HPLC). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), AST/ALT, total bilirubin (TBIL), direct bilirubin (DBIL) and indirect bilirubin (IBIL) were detected with automatic biochemical analyzer. The median concentrations of serum NP, BPA, and DMP in the participants were 45.85 ng/mL, 26.31 ng/mL and 31.62 ng/mL, respectively, which were higher than the environmental concentration limits of the U.S. National Environmental Protection Agency (EPA). Hair gels used during pregnancy, types of domestic drinking water, nail polish and cosmetics used by children were significantly positive correlated with serum NP concentration (P < 0.05). Gender, feeding pattern, plastic water cup used during pregnancy, hair spray and perfume use for children, duration of children birth, materials for baby bottle or cup and ways to plastic products were significantly positively correlated with serum BPA concentration (P < 0.05). Gender, perms used during pregnancy, hair spray and perfume use for children, using plastic lunch box during pregnancy, duration of children birth, exposure to pesticides, parents' occupations were significantly positively correlated with serum DMP concentrations (P < 0.05). Serum NP (β = 0.296, P = 0.036) and DMP (β = 0.316, P = 0.026) concentrations and TBIL level were significantly positively correlated. Serum NP concentration and the levels of IBIL (β = 0.382, P = 0.006) are significantly positively correlated. Cosmetics used during pregnancy significantly increased AST level (β = 2.641, P = 0.021). There was a positive correlation between the frequency of hair spray and perfume use for children and the AST (β = 4.241, P = 0.022). NP, BPA and DMP, which were commonly detected in the serum of children aged 5-14 years old in Zunyi City, Northern Guizhou Province, China, were closely related to the environmental risk factors of exposure environment during pregnancy, infancy and school age. Exposure to NP, BPA and DMP would have negative effects on hepatic function, and these effects showed differences in gender and geographical location. Notably,The relationships were more evident in girls than in boys.

Background & Aims: Anti-gp210 antibodies are highly specific for primary biliary cholangitis (PBC) and are detected in 20%-40% of PBC patients. PBC patients with anti-gp210 positive appear to have more advanced bile duct damage and worst prognosis. Recent studies suggest that miRNAs play a critical role in the progression of PBC. The aim of this study was to explore the relationship between the expression dysregulation of miRNAs in serum from patients with anti-gp210 positive PBC and the disease progression. Methods: The differentially expressed miRNAs were identified by deep sequencing. The differentially expressed miRNAs were validated by qRT-PCR in PBC patients along with healthy controls and chronic hepatitis B patients. The target genes of differentially expressed miRNAs were predicted using TargetScan and miRDB database. Gene ontology and KEGG pathway analyses were conducted to explore the target genes function. Results: The expression levels of 60 miRNAs were found to differ significantly between PBC patients with anti-gp210 positive and anti-gp210 negative. MiR-410-3p was validated as the significantly differentially expressed miRNA. MiR-410-3p had a higher level in patients with anti-gp210 positive PBC and was associated with the disease progression. Bioinformatics showed that underlying target genes of miR-410-3p was involved in cell communication, signal transduction, cell apoptosis, and so on. Through KEGG pathway analysis, we suspected that Th1, Th2, Th17 cell differentiation might be associated with the pathogenesis of PBC. Conclusion: The over-expression of serum miR-410-3p might reflect the disease progression in patients with anti-gp210 positive PBC.

It has been suggested that the heterogeneity of TAAs in lung cancer may affect the therapeutic response and disease progression. Up to now, several tumor-associated antigen (TAA)-based cancer vaccines have been investigated in lung adenocarcinoma (LUAD); however, most of them have failed at the stage of clinical trials. The present study suggests that inter-tumoral heterogeneity of TAAs is large at single-cell resolution, while the communication between tumor cells and infiltrating T cells is closely related to the expression profile of TAAs.

The aim of this work was to determine whether chronic exposure to nonylphenol (NP), a representative substance of environmental endocrine disruptors (EEDs), at environmental concentration would have toxic effects on thyroid function and thyroid hyperplasia disease. Two hundred SPF Sprague-Dawley rats were divided into five groups (n = 40 per group): blank control group (corn oil), low-dose NP exposure group (0.4 mg/kg/d), medium-dose NP exposure group (4 mg/kg/d), high-dose NP exposure group (40 mg/kg/d), and estradiol control group (E2: 30 μg/kg/d). The rats were treated by gavage for 34 weeks, which were sampled twice (17 weeks and 34 weeks respectively). NP accumulation in the thyroid tissue (F = 52.93, P < 0.001) and serum (F = 5.54, P = 0.00) continuously increased in a significant dose-effect relationship. After NP exposure, the serum FT3 levels exhibited a dose-dependent increasing trend (F = 4.68, P = 0.01), while the serum FT4 level showed an opposite trend (F = 3.93, P= 0.01). Compared with the control group, hyperechoic areas (i.e., calcification points) were observed in the high-dose group. Follicular epithelial stratification was extremely severe, the monolayer cubic epithelial cells became flat, and the area of single follicles was even smaller in the high-dose group. In the high-dose NP group, there were numerous mitochondria that were severely swollen. The rough endoplasmic reticulum was abundant, with obvious expansion and vesiculation. The relative expression of ERα (F = 5.29, P = 0.00), ERβ (F = 10.17, P = 0.00), TRα (F = 7.71, P = 0.00), TRβ (F = 3.52.17, P = 0.02) and HMGB1 (F = 10.16, P = 0.01) proteins in the thyroid tissue in each NP exposure group was increased compared with the control group, and the relative expression of proteins increased if the exposure time was prolonged under the same exposure dose. Chronic exposure to NP at environmental concentration could have toxic effects on thyroid function, and induce thyroid hyperplasia disease in male rats.

Accumulating evidence suggests that the malignant phenotypes of cancers are determined not only by the intrinsic properties of cancer cells but also by components in the tumor microenvironment (TME). In this study, we comprehensively characterized the TME of cutaneous melanoma (CM). As a result, tumor stage, tissue site, ulceration, thickness as well as patient age, sex were associated with immune infiltration. Patients of higher immune infiltration exhibited better survival outcomes, and antitumor effector cells, such as CD8 T cells and M1 macrophages, were found in significantly higher numbers in those tissues. Differential expression of mRNAs and long non-coding RNAs (lncRNAs) was analyzed and utilized to construct an immune-related competing endogenous RNA network, in which a lncRNA-associated subnetwork that could positively regulate the expression of IFN-γ was highlighted. Functional analysis confirmed that this network was remarkably enriched in functional terms related to both immune response and tumor-intrinsic pathways. Finally, a total of 109 high-confidence prognostic genes were identified, and a gene module that contained several key immune checkpoint molecules or modulators (PD-1, PD-L1, PD-L2, and LCK) was screened, which confers survival benefit for CM patients as supported by both overall and relapse-free survival rates from different datasets.

Background and aim A nonskeletal role of vitamin D in primary biliary cholangitis (PBC) patients is not yet clear. The purpose of this study is to investigate the serum vitamin D level in patients with PBC and to explore whether serum vitamin D level is related to disease progression. Methods The serum levels of 25(OH)D(3)were measured in 185 PBC patients and 141 healthy controls. The association with vitamin D levels and disease progression, particularly natural course, was assessed. Results Serum vitamin D levels were measured in 185 patients with PBC. The mean level of vitamin D in advanced stage patients was lower than that in early stage patients (9.15 +/- 5.33 ng/ml vs. 13.68 +/- 6.33 ng/ml,p = .000). In the follow-up patients, the vitamin D level in the taken calcitriol patients showed an upward trend, while the un-taken calcitriol patients was opposite. Besides, vitamin D levels were correlated with the changes of bilirubin, albumin (ALB) and APRI (p<.05). PBC patients with vitamin D deficiency had higher bilirubin levels and lower ALB levels (p<.05). Moreover, there were differences in serum vitamin D levels between taken calcitriol patients and un-taken calcitriol patients (p = .027). Conclusions Vitamin D deficiency is highly prevalent in PBC patients. The decrease of vitamin D levels may indicate disease progression in PBC. PBC patients need to be regularly measured for serum vitamin D level and take appropriate vitamin D supplementation.

BACKGROUND:To identify serum nonylphenol (NP) and glucolipid metabolism-related proteins in Type 2 diabetes (T2D) patients.METHODS:We performed a hospital-based, case-control study in patients admitted to the Department of Endocrinology, Hospital of Zunyi Medical University, Zunyi City, China from Mar to Nov of 2014. The study included 112 T2D cases diagnosed in accordance with the 2013 WHO Expert Committee Diabetes Diagnosing Criteria, and 125 healthy individuals with normal fasting blood glucose (FBG) when receiving physical examination in the same period in the Municipal Physical Examination Center. Blood samples from subjects in the 2 groups underwent detection of biochemical indices, including FBG, blood fat, and NP. Glucolipid metabolism-related proteins, including estrogen receptor (ER), sterol regulatory element-binding protein-1c (SREBP-1c), wingless-type MMTV integration site family member 5a (Wnt5a), and peroxisome proliferator-activated receptor-γ (PPAR-γ). These indices were compared between the 2 groups to analyze the correlation between serum NP levels and glucolipid metabolic proteins.RESULTS:The subjects in the diabetes group had higher triglycerides (TG), total cholesterol (TC), NP, ER, SREBP-1c, Wnt5a, FBG, and TG levels than the healthy group, but lower levels of low-density lipoprotein cholesterol (LDL-C) and PPAR-γ than the healthy group. No significant differences in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found between the two groups. The serum NP levels were shown to be positively correlated with SREBP-1c but negatively correlated with PPAR-γ.CONCLUSION:The serum NP levels of T2D patients is higher than the levels in healthy controls, and its levels correlate with SREBP-1c and PPAR-γ levels.

The protective effect of zinc selenium tea against metabolic syndrome (MetS) was tested by using a high-sucrose-high-fat diet (HSHFD)-induced MetS model. Fifty Sprague–Dawley rats were randomly divided into five groups: normal diet (C-group), HSHFD (CH-group), HSHFD + green tea (0.24 g/kg/day) (TH-group), HSHFD + low-dose zinc selenium organic tea (0.24 g/kg/day) (ZTHL-group), and HSHFD + high-dose zinc selenium organic tea (1.20 g/kg/day) (ZTHH-group). After 8 weeks, compared to both the C-group and CH-group, the hepatosomatic index (HI) was significantly reduced in the ZTHL-group (p < 0.05). Fasting blood glucose (FBG) levels were highest in the TH-group, followed by the CH-group, then the ZTHL-group, then the ZTHH-group, and finally the C-group. Compared with the CH-group, the serum total cholesterol (TC) and low density lipid-cholesterol (LDL-C) concentrations were significantly lower in the ZTHH-group (p < 0.05). Significant decreases in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acids (TBA), alkaline phosphatase (ALP), and direct bilirubin (DBIL) levels were observed in ZTHL-group versus the CH-group (p < 0.05). Serum alpha-L-fucosidase (AFU) levels in the ZTHH-group were lower than in the CH-group (P < 0.01). Histopathological examination of the liver and fat biopsies illustrates that the liver cells showed a decrease in the extent of necrosis and dropsy in the ZTHL-group and ZTHH-group versus the CH-group. Zinc selenium tea showed a protection effect against hepatic damage.

Exposure to environmental endocrine disruptors (EEDs) contributes to the pathogenesis of many metabolic disorders. Here, we have analyzed the effect of the EED-nonylphenol (NP) on the promotion of non-alcoholic fatty liver disease (NAFLD) in rats fed high sucrose-high fat diet (HSHFD). Fifty Sprague-Dawley rats were divided into five groups: controls fed a normal diet (C-ND); HSHFD-fed controls (C-HSHFD); and rats fed a HSHFD combined with NP at doses of 0.02 μg/kg/day (NP-L-HSHFD), 0.2 μg/kg/day (NP-M-HSHFD), and 2 μg/kg/day (NP-H-HSHFD). Subchronic exposure to NP coupled with HSHFD increased daily water and food intake ( p < 0.05), hepatic echogenicity and oblique liver diameter ( p < 0.05), and plasma levels of alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglycerides, and low density lipoprotein cholesterol ( p < 0.05). Combined exposure to NP and HSHFD induced macrovesicular steatosis with dilation and congestion of the central vein, liver inflammatory cell infiltration, and expression of genes regulating lipid metabolism, SREBP-1C, FAS, and Ucp2. These results demonstrate that NP aggravates NAFLD in HSHFD-treated rats by up-regulating lipogenic genes, and that HSHFD increases the toxic effects of NP. Thus subchronic NP exposure may lead to NAFLD, especially when combined with a high-sucrose/high-fat diet.

OBJECTIVES:To investigate the effects of Nonylphenol (NP) in pups from dams exposed during gestational and lactational periods on immediate early genes (c-jun, c-fos) in hippocampus and the learning and memory of F1 rats. MATERIALS AND METHODS:Twenty eight pregnant dams, stratified by pregnancy date, were randomly assigned into 4 groups, which were gavaged with NP at the doses of 50 mg/kg/day, 100 mg/kg/day, 200 mg/kg/day and groundnut oil, respectively. Step-down avoidance test, and learning and memory effects of NP were evaluated on 8-weeks-old pups. The expressions of c-jun and c-fos and the activities of choline acetyltransferase (ChAT) and acetylcholinesterase (AchE) were evaluated in hippocampus of pups. RESULTS:Compared to the control, reaction time (RT) that pups spent to jump to the platform was longer (P=0.02), the number of errors were higher (P=0.01), and the step-down latency was shorter in the 200 mg/kg/day NP-treated group (P=0.04). Exposure to NP induced a significant reduction in ChAT activity in hippocampus in the 100 (P=0.005) and 200 mg/kg/day NP-treated groups (P=0.002), whereas exposure to 200 mg/kg/day caused a significant increase in AchE activity (P=0.004); a dose-response relationship was revealed between ChAT and AchE activities and NP exposure in the hippocampus of pups (r=-0.821, P=0.01; r=0.757, P=0.04). Exposure to NP in the 100 and 200 mg/kg/day NP-treated groups exhibited an increase in number of c-fos and c-jun positive cells. CONCLUSION:Exposure to NP might negatively affect learning and memory ability in F1 rats, possibly due to the alterations in the expression of c-jun and c-fos, and ChAT, AchE activities in hippocampus of pups.

Objective To observe the effect of nonylphenol (NP) exposure on offspring weight,fat generation and adipocyte differentiation genes,liver and adipocyte changes of pregnant rats induced by high-fat and high-sugar diet during perinatal period and to explore the mechanism of offspring obese occurrence via high-fat and high-glucose diet treatment combined with NP exposure in perinatal mothers.Methods Forty female SD rats were mated with 20 male SD rats.Then we collected the pregnant rats and divided them into the ordinary diet control group (OC),The high fat diet control group (HC),high fat+low-dose Nonylphenol group (HNL),and high fat + high-dose Nonylphenol group (HNH).Rats in the OC grup were fed with normal diet,and rats in the HC,HNL and HNH groups were fed with high fat and high sugar diet.Pregnant rats were given gestational gavage from day 6 to postpartum 21 days.In the OC group and HC group,the pregnant rats were given 0.5 mL/(kg·d) corn oil,in the HNL group and HNH,they were given NP 50,100 mg/(kg·d) by gavage.Pregnant rats were fed with normal breast feeding and fed with normal diet for up to 70 days after birth.After 70 days of intervention,the blood of the abdominal aorta of the offspring was taken and the levels of serum TG,TC,HDL and LDL were measured by automatic biochemical analyzer.Before taking blood,we measured the weight of offspring rats.After taking the blood,we took the gonadal fat pad to measure fat quality and calculate the fat coefficient.We collected the liver tissues of the offsprings,Using real-time PCR to detect the regulation of adipocyte differentiation key gene transcription factor (CEBP-α),sterol regulatory binding protein (SREBP-1),peroxisome proliferator-activated receptor γ (PPARγ) mRNA expression.Liver tissues and adipose tissues were observed by HE staining.Results The levels of serum TG,TC and LDL in the HNL group and HNH group were higher than those in the HC group (all P<0.01).The body mass and fat coefficient of the HNL group and HNH group were higher than those of the HC group,and the fat quality of the HNH group was higher than that of the HC group (all P<0.01).The expression of CEBP-α mRNA in the HNH group was higher than that in the OC group (P<0.01).The expression of SREBP-1 and PPARγ mRNA in the HNH group was higher than that in the HC group (all P<0.01).HE staining showed that some liver cells were steatosis in the HC group,and a large number of hepatocyte fat-like changes were seen in the HNL group.The extensive balloon-like and large amount of lipid droplets were found in the HNL group.In the HC group,part of the fat cell area increased,and in the HNL group,most of the fat cell area increased significantly.Conclusion High-fat and high-glucose diet combined with NP exposure on pregnant rats can increase the offspring body mass,fat quality and blood lipid levels,which lead to increasing the liver fat and adipocytes,and the mechanism may be that NP leads to the expression abnormalities of adipocyte differentiation genes CEBP-α,SREBP-1,and PPARγ.