Interleukin-17A educated hepatic stellate cells promote hepatocellular carcinoma occurrence through fibroblast activation protein expression.

BACKGROUND: Liver cancer is typified by a complex inflammatory tumor microenvironment, where an array of cytokines and stromal cells orchestrate a milieu that significantly influences tumorigenesis. Interleukin-17A (IL-17A), a pivotal pro-inflammatory cytokine predominantly secreted by Th17 cells, is known to play a substantial role in the etiology and progression of liver cancer. However, the precise mechanism by which IL-17A engages with hepatic stellate cells (HSCs) to facilitate the development of hepatocellular carcinoma (HCC) remains to be fully elucidated. This investigation seeks to unravel the interplay between IL-17A and HSCs in the context of HCC. METHODS: An HCC model was established in male Sprague-Dawley rats using diethylnitrosamine to explore the roles of IL-17A and HSCs in HCC pathogenesis. In vivo overexpression of Il17a was achieved using adeno-associated virus. A suite of molecular techniques, including RT-qPCR, enzyme-linked immunosorbent assays, Western blotting, cell counting kit-8 assays and colony formation assays, was employed for in vitro analyses. RESULTS: The study findings indicate that IL-17A is a key mediator in HCC promotion, primarily through the activation of hepatic progenitor cells (HPCs). This pro-tumorigenic influence appears to be mediated by HSCs, rather than through a direct effect on HPCs. Notably, IL-17A-induced expression of fibroblast activation protein (FAP) in HSCs emerged as a critical factor in HCC progression. Silencing Fap in IL-17A-stimulated HSCs was observed to reverse the HCC-promoting effects of HSCs. CONCLUSIONS: The collective evidence from this study implicates the IL-17A/FAP signaling axis within HSCs as a contributor to HCC development by enhancing HPC activation. These findings bolster the potential of IL-17A as a diagnostic and preventative target for HCC, offering new avenues for therapeutic intervention.

IL-17a promotes hepatocellular carcinoma by increasing FAP expression in hepatic stellate cells via activation of the STAT3 signaling pathway.

Studies have shown that hepatic stellate cells (HSCs) and interleukin-17a (IL-17a) play important roles in liver tumorigenesis. In addition, fibroblast activation protein-α (FAP) has been shown to be a key regulator of hepatic stellate cell activation. In this study, in vivo and in vitro experiments were performed to verify the promoting effects of IL-17a administration, IL-17a overexpression, and FAP upregulation in HSCs on liver fibrosis and liver tumorigenesis. The cleavage under targets & release using nuclease (CUT&RUN) technique was used to verify the binding status of STAT3 to the FAP promoter. The in vitro studies showed that IL-17a activated HSCs and promoted HCC development and progression. FAP and IL-17a overexpression also activated HSCs, promoted HCC cell proliferation and migration, and inhibited HCC cell apoptosis. The in vivo studies suggested that IL-17a and FAP overexpression in HSCs facilitated liver tumor development and progression. The CUT&RUN results indicated that FAP expression was regulated by STAT3, which could bind to the FAP promoter region and regulate its transcription status. We concluded that IL-17a promoted HCC by increasing FAP expression in HSCs via activation of the STAT3 signaling pathway.

High-throughput sequencing approach for the identification of lncRNA biomarkers in hepatocellular carcinoma and revealing the effect of ZFAS1/miR-150-5p on hepatocellular carcinoma progression.

Aims: To screen abnormal lncRNAs and diagnostic biomarkers in the progression of hepatocellular carcinoma through high-throughput sequencing and explore the underlying mechanisms of abnormal lncRNAs in the progression of hepatocellular carcinoma. Methods: The transcriptome sequencing was used to analyze the RNA expression profile and identify differentially expressed RNAs. Hub lncRNAs were screened by combining (WGCNA, ceRNA regulatory network, PPI, GO and KEGG analyses, Kaplan-Meier curve analysis, Cox analysis, risk model construction and qPCR). Thereafter, the correlation between the expression of hub lncRNAs and tumor clinicopathological parameters was analyzed, and the hub lncRNAs were analyzed by GSEA. Finally, the effects of hub RNAs on the proliferation, migration and invasion of HepG2 cells were investigated in vitro. Results: Compared with the control group, a total of 610 lncRNAs, 2,593 mRNAs and 26 miRNAs were screened in patients with hepatocellular carcinoma. Through miRNA target prediction and WGCNA, a ceRNA was constructed, comprising 324 nodes and 621 edges. Enrichment analysis showed that mRNAs in ceRNA were involved mainly in cancer development progression. Then, the ZFAS1/miR-150-5p interaction pair was screened out by Kaplan Meier curve analysis, Cox analysis and qPCR analysis. Its expression was related to tumor stage, TNM stage and patient age. ROC curve analysis showed that it has a good predictive value for the risk of hepatocellular carcinoma. GSEA showed that ZFAS1 was also enriched in the regulation of immune response, cell differentiation and proliferation. Loss-of-function experiments revealed that ZFAS1 inhibition could remarkably suppress HepG2 cell proliferation, migration and invasion in vitro. Bioinformatic analysis and luciferase reporter assays revealed that ZFAS1 directly interacted with miR-150-5p. Rescue experiments showed that a miR-150-5p inhibitor reversed the cell proliferation, migration and invasion functions of ZFAS1 knockdown in vitro. Conclusion: ZFAS1 is associated with the malignant status and prognosis of patients with hepatocellular carcinoma, and the ZFAS1/miR-150-5p axis is involved in hepatocellular carcinoma progression.

Research on Fault Diagnosis of Rolling Bearings Based on Variational Mode Decomposition Improved by the Niche Genetic Algorithm.

Due to the influence of signal-to-noise ratio in the early failure stage of rolling bearings in rotating machinery, it is difficult to effectively extract feature information. Variational Mode Decomposition (VMD) has been widely used to decompose vibration signals which can reflect more fault omens. In order to improve the efficiency and accuracy, a method to optimize VMD by using the Niche Genetic Algorithm (NGA) is proposed in this paper. In this method, the optimal Shannon entropy of modal components in a VMD algorithm is taken as the optimization objective, by using the NGA to constantly update and optimize the combination of influencing parameters composed of α and K so as to minimize the local minimum entropy. According to the obtained optimization results, the optimal input parameters of the VMD algorithm were set. The method mentioned is applied to the fault extraction of a simulated signal and a measured signal of a rolling bearing. The decomposition process of the rolling-bearing fault signal was transferred to the variational frame by the NGA-VMD algorithm, and several eigenmode function components were obtained. The energy feature extracted from the modal component containing the main fault information was used as the input vector of a particle swarm optimized support vector machine (PSO-SVM) and used to identify the fault type of the rolling bearing. The analysis results of the simulation signal and measured signal show that: the NGA-VMD algorithm can decompose the vibration signal of a rolling bearing accurately and has a better robust performance and correct recognition rate than the VMD algorithm. It can highlight the local characteristics of the original sample data and reduce the interference of the parameters selected artificially in the VMD algorithm on the processing results, improving the fault-diagnosis efficiency of rolling bearings.

Probabilistic ratiocination of hepatocellular carcinoma after resection: evaluation of expected to be promising approaches.

BACKGROUND: Precise prediction of survival after treatment is of great importance for patients with diseases with high mortality. RNA sequencing data and deep learning (DL) methods are expected to become promising approaches in the development of prediction models in the future. We aimed to evaluate the optimal covariates and methodology for patients with hepatocellular carcinoma (HCC) undergoing surgical resection. METHODS: The Cox proportional hazards regression model and the DL approach were used to develop prediction models incorporating clinical, genetic, and combined clinical and genetic variables for survival prediction in patients with HCC after resection. A total of 1,114 patients and 184 patients were enrolled in the present study from 2,163 and 601 patients from Eastern Hepatobiliary Surgery Hospital and Renji Hospital, respectively. The models were internally validated through random sampling and externally validated in clinical cohorts. Between-model comparisons were carried out in terms of the integrated discrimination improvement and net reclassification index. RESULTS: The Cox and DL clinical models were developed by adopting 7 independent prognostic factors (total bilirubin, prothrombin time, tumor size, tumor number, lymph node metastasis, and vascular invasion) and 22 clinical factors, respectively. Both the Cox clinical model and the DL clinical model showed excellent performances in the derivation [area under the curve (AUC): 0.75 vs. 0.77] and validation (AUC: 0.83 vs. 0.80) sets. The derived Cox genetic model with 6 significant prognostic genes was not as effective as the DL approach involving 686 genes. A combined clinical and genetic approach modified the performances of both the Cox and DL models. The integrated discrimination improvement and net reclassification index of the DL clinical model were generally better than those of the Cox clinical model. CONCLUSIONS: Our Cox clinical model sufficiently provided precise survival prediction in patients with HCC after resection. It may serve as an accurate and cost-effective tool for predicting survival in such patients.

Self-harm behaviors, suicidal ideation, and associated factors among rural left-behind children in west China.

PURPOSE: This population-based cross-sectional study describes self-harm (SH) behaviors and associated factors in 2898 Chinese left-behind children (LBC). METHODS: Descriptive statistics were used to present and compare the distributional characteristics of SH behaviors. Binary and ordinal logistic-regression models for survey data were applied to explore associated factors of SH. RESULTS: Among the LBC, 48% (95% CI: 42%-54%) reported SH behaviors. SH was prominently associated with suicidal ideation. Boys, junior high-school students, and LBC whose fathers were poorly educated were observed to have higher prevalence of SH. Boys, local ethnic minorities, junior high-school students, and LBC whose fathers were comparatively well educated were observed to be at increased risk of more medically serious SH behaviors. CONCLUSIONS: SH is a substantial public health problem among rural Chinese LBC. Targeted intervention measures should be developed to reduce future suicide risk.

Antiviral therapy improves post-operative survival outcomes in patients with HBV-related hepatocellular carcinoma of less than 3 cm - A retrospective cohort study.

BACKGROUND: The impact of antiviral therapy on long-term survival outcomes in patients with small HBV-related hepatocellular carcinoma (HBV-related HCC) after liver resection is still controversial, as the impact can be overshadowed by tumor-related factors. This study investigated this impact on recurrence and survival in patients with HCC of less than 3 cm. OBJECTIVE: This study was designed to further determine the impact of antiviral treatment on prognosis of patients with HCC after liver resection, to verify whether patients with cirrhosis still benefited from antiviral treatment, to study the impact of antiviral treatment on post-operative HCC recurrence, and to determine whether patients with a low preoperative HBV-DNA viral load should receive antiviral therapy. METHODS: The clinical data on patients who underwent curative liver resection for histopathologically confirmed small HCC (≤3 cm in diameter) were analyzed to determine factors which were related with HCC recurrence and survival. The disease-free and overall survival outcomes were estimated by the Kaplan-Meier method. Univariate and multivariate analyses were performed to identify the risk factors of long-term survival. RESULTS: Of the 795 patients in this study, patients with high preoperative HBV-DNA levels had significantly worse DFS and OS outcomes at 1-, 3- and 5- year after liver resection when compared with those with low HBV-DNA levels (86.1%, 60.8%, 46.6% vs 90.5%, 71.3%, 51.4%; and 98.5%, 89.3%, 75.2% vs 98.8%, 91.5%, 84%, respectively). Patients who received antiviral therapy had significantly better DFS and OS outcomes at 1-, 3- and 5- year after liver resection when compared with those without (91.6%, 69.5%, 55% vs 80.2%, 56%, 44.2%; and 99.6%, 93.5%, 87% vs 96.1%, 80.5%, 61.3%, respectively). Antiviral therapy significantly improved the OS but not DFS outcomes in patients with low HBV-DNA levels. The corresponding 1-, 3- and 5- year DFS and OS outcomes were 92.6%, 73%, 59.1% vs 87.1%, 68.5%, 57.9%; and 99.5%, 95.1%, 91.1% vs 97.6%, 85.5%, 72.4%, respectively. Antiviral treatment significantly prolonged DFS and OS in patients with cirrhosis. The corresponding 1-, 3- and 5- year DFS and OS were 90.2%, 66%, 49% vs 73.9%, 46.6%, 32.8%; and 100%, 93.6%, 85% vs 93.8%, 73.3%, 52.6%, respectively. CONCLUSION: Antiviral therapy improved the prognosis of small HBV-related HCC of less than 3 cm. The survival benefit was also detected in patients with cirrhosis. Antiviral therapy should be considered a routine post-operative therapy for patients with HBV-related HCC.

MicroRNA-520e targets AEG-1 to suppress the proliferation and invasion of colorectal cancer cells through Wnt/GSK-3ß/ß-catenin signalling.

MicroRNA-520e (miR-520e) is increasingly being recognized as a cancer-related miRNA in multiple cancer types; however, little is known about its role in colorectal cancer. In this study, we determined the specific role of miR-520e in colorectal cancer. Expression of miR-520e was lower in colorectal cancer tissues compared to normal tissues. Overexpression of miR-520e significantly decreased the proliferation, colony formation and invasion of colorectal cancer cells, while inhibition of miR-520e exhibited the opposite effect. Moreover, miR-520e was found to target the 3'-untranslated region of astrocyte elevated gene-1 (AEG-1) and inhibit AEG-1 expression in colorectal cancer cells. An inverse correlation between miR-520e and AEG-1 expression was confirmed in colorectal cancer tissues. Notably, miR-520e suppressed the phosphorylation of glycogen synthase kinase-3ß and decreased the expression of ß-catenin, leading to inactivation of Wnt/ß-catenin signalling in colorectal cells. A rescue assay confirmed that miR-520e regulates cell proliferation, invasion and Wnt/ß-catenin signalling through targeting AEG-1. Taken together, these results indicate that miR-520e plays a critical role in regulating colorectal cancer cell proliferation and invasion by inhibiting Wnt/ß-catenin signalling via AEG-1. Our study highlights the importance of the miR-520e/AEG-1/Wnt/ß-catenin signalling axis in colorectal cancer, thus targeting miR-520e may represent an effective therapeutic strategy.

Early clinical management of acute CNS infections: a prospective cohort study.

Aim: To explore early management and clinical predictors of patients with suspected CNS infections. Methods: In a prospective cohort study of 125 adult patients with suspected CNS infections, clinical features and early management time points were compared between groups with and without confirmed CNS infections. Results: The door-to-lumbar puncture time was associated with the initial Glasgow Coma Scale score, the confirmed diagnosis and the time to change empirical treatment. Multivariate analysis indicated that the initial Glasgow Coma Scale score was an independent risk factor for prognosis. Conclusion: Lumbar puncture plays a crucial role in early management of CNS infections. Patients with CNS infection who have disturbances of consciousness should receive particular attention.

Effect of Surface Topography Parameters on Friction and Wear of Random Rough Surface.

In order to explore the relationship between the surface topography parameters and friction properties of a rough contact interface under fluid dynamic pressure lubrication conditions, friction experiments were carried out. The three-dimensional surface topography of specimens was measured and characterized with a profile microscopy measuring system and scanning electron microscope. The friction coefficient showed a trend of decreasing first and then increasing with the increase in some surface topography parameters at lower pressure, such as the surface height arithmetic mean Sa, surface height distribution kurtosis Sku, surface volume average volume Vvv, and surface center area average void volume Vvc, which are the ISO 25178 international standard parameters. The effects of surface topographic parameters on friction were analyzed and the wear mechanism of the worn surface was presented. The wear characteristics of the samples were mainly characterized as strain fatigue, grinding, and scraping. The results provide a theoretical basis for the functional characterization of surface topography.

MiR-299-3p functions as a tumor suppressor via targeting Sirtuin 5 in hepatocellular carcinoma.

Hepatocellular carcinoma(HCC) is one of the most common cancers in the world, with the characteristics of high morbidity and mortality. Though the levels of diagnosis and treatment of HCC have been largely improved recently, the prognosis of these patients remains unacceptable. Thus, it is urgent for us to discover the exact mechanisms and determine some new biomarkers for HCC. Previous studies revealed that microRNAs (miRNAs) played a critical role in the occurrence and progression of HCC. And miR-299-3p has been reported to be closely related to the progression of colon carcinoma, lung cancer and clear cell renal cell carcinoma and so on. However, the exact expression and functions of miR-299-3p in HCC are still uncovered. Here, we reported for the first time that miR-299-3p was downregulated in HCC. Clinically, statistical analysis showed that miR-299-3p expression was significantly associated with tumor size (P = 0.007), venous infiltration (P = 0.028), Edmondson-Steiner grading (P = 0.042) and TNM stage (P = 0.012). In addition, HCC patients with lower miR-299-3p expression had worse 3-year overall survival and disease-free survival (P = 0.0012, P = 0.0002). Functionally, Transwell assays, Wound healing assay, MTT assay and plate clone formation assay revealed that miR-299-3p inhibited the migration, invasion and proliferation of HCC cells. Furthermore, bioinformatics tools, luciferase reporter assay, real-time PCR, Pearson's correlation coefficient analysis, immunohistochemistry and Western blot showed that Sirtuin 5 (SIRT5) was a downstream target of miR-299-3p in HCC cells. In addition, rescue experiments indicated that SIRT5 mediated the effects of miR-299-3p on migration, invasion and proliferation of HCC cells. Thus, we conclude that miR-299-3p suppresses migration, invasion and proliferation of HCC cells via directly targeting SIRT5. MiR-299-3p may be a potential prognosis indicator and therapeutic target for HCC.

Paraoxonase 3 inhibits cell proliferation and serves as a prognostic predictor in hepatocellular carcinoma.

Paraoxonase 3 (PON3) exerts prominent anti-inflammation and anti-oxidation properties mainly at the cellular level, and is primarily expressed in the liver. However, its role in HCC remains unexplored. Here, we investigated the expression pattern, clinical significance, and function of PON3 in HCC. PON3 mRNA and protein levels were respectively determined in two large cohorts using quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) of tissue microarray. We found that PON3 was downregulated in most HCCs. Kaplan-Meier and log-rank test showed that PON3 downregulation predicted shorter recurrence-free survival (RFS) and overall survival (OS) time in all HCC patients, especially early-stage HCC patients. Cox regression analysis revealed that the PON3 downregulation was an independent risk factor for RFS and OS. Gain- and loss-of-function experiments revealed that PON3 suppressed cell proliferation in vivo and in vitro, which was attributed to its cell-cycle arrest effect. In addition, microarray analysis showed that some pro-proliferative genes were elevated when PON3 was knockdown, and these genes possibly involved in the underlying mechanisms. In conclusion, our studies reveal the cell proliferation inhibitory function of PON3 and offer a potential prognostic predictor and therapeutic target for HCC.

[Camostat mesilate, a protease inhibitor, inhibits visceral sensitivity and spinal c-fos expression in rats with acute restraint stress].

OBJECTIVE: To observe the effect of gut protease activity on visceral hypersensitivity in rats with acute restraint stress. METHODS: Sprague-Dawley rats were given 30, 100 or 300 mg/kg camostat mesilate (CM), a protease inhibitor, or saline intragastrically 30 min before acute restraint stress induced by wrapping the fore shoulders, upper forelimbs and thoracic trunk for 2 h. Visceral perception of the rats was quantified as the visceral motor response with an electromyography, and the rectal mucosa and feces protease activity and spinal c-fos expression were measured. RESULTS: CM dose-dependently reduced visceral sensitization elicited by rectal distension, but these doses did not completely inhibit stress-induced visceral sensitization. In normal rats, c-fos expression was found mainly in the superal spinal cord dorsal horn, and after the administration the CM, c-fos-positive cells decreased significantly in all dose groups (P<0.05). In 30 mg/kg CM group, fecal and rectal mucosal protease activity significantly decreased as compared with that in the stress group (P<0.05), and as CM dose increased to 100 and 300 mg/kg, the protease activity decreased even further (P<0.01). CONCLUSION: The gut protease is involved in acute stress-induced visceral hypersensitivity, and CM can lower the visceral sensitivity and spinal c-fos expression in rats.

The expression of protease-activated receptor 2 and 4 in the colon of irritable bowel syndrome patients.

BACKGROUND: Irritable bowel syndrome (IBS) pathophysiology has not been fully understood. Abnormalities of serine proteases have been identified in IBS patients. In addition, protease-activated receptors (PAR) activation interferes with several components of the pathogenesis of IBS, so, evaluating the PAR expression in IBS patients may contribute to understanding the pathogenesis of the disease. AIMS: This study aimed to investigate whether the expression of PAR4 and PAR2 in the colon was changed in IBS patients and was associated with IBS. METHODS: Colon mucosal biopsies of 34 IBS patients (16 constipation- and 18 diarrhea-predominant) and 18 control subjects were collected. Gene transcripts of PAR2, PAR4, tryptase, and trypsin were quantified using real-time polymerase chain reaction (PCR) and the expression of PAR2 and PAR4 receptors was also measured by immunohistology and image analysis. RESULTS: In IBS patients, the mRNA expression of tryptase and trypsin normalized against ß-actin gene was higher compared to control subjects (P < 0.001). No difference was observed in the PAR2 mRNA level or protein level between control subjects on the one hand and IBS patients or subgroups on the other. In IBS or IBS subgroups patients, the expression of PAR4 in the mRNA level or protein level was lower than the control subjects. CONCLUSIONS: This study, for the first time, indicated the PAR4 expression in IBS patients. Decreased PAR4 expression may help us to understand the pathogenesis of IBS.

A protease inhibitor against acute stress-induced visceral hypersensitivity and paracellular permeability in rats.

In the present study, we investigated the effects of camostat mesilate (CM), a synthetic protease inhibitor, on visceral sensitivity and paracellular permeability induced by the acute restraint stress. We also explored the possible mechanisms underlying these effects. The acute restraint stress was induced by wrapping the fore shoulders, upper forelimbs and thoracic trunk of Sprague-Dawley rats for 2h. Either CM (30, 100 and 300mg/kg) or saline was intragastrically administrated to the rats 30min before the acute restraint stress. Visceral perception was quantified as visceral motor response with an electromyography in a subset of rats. Paracellular permeability was determined in another subset of rats. We found that the visceral sensitivity and paracellular permeability were significantly reduced in the CM-treated rats. Moreover, the fecal protease activity was decreased in the CM-treated rats. The ZO-1 protein expression was markedly reduced by the stress treatment, but this decrease was suppressed by CM administration. Our data indicated that CM could efficiently inhibit visceral sensitivity and paracellular permeability induced by the acute restraint stress in rats. Therefore, CM might be an effective drug for the treatment of irritable bowel syndrome.