Effect and potential mechanism of oncometabolite succinate promotes distant metastasis of colorectal cancer by activating STAT3.

To investigate the effect of Oncometabolite succinate on colorectal cancer migration and invasion and to initially explore the underlying mechanism.Succinate acid detection kit detected the succinate content in tissues. The growth of colorectal cancer cells was measured by cck-8 assay, wound-healing migration assay and transwell migration and invasion assays, and then explored the level of epithelial-mesenchymal transition (EMT) and STAT3/ p-STAT3 expression by western blot analysis and quantitative real-time PCR for mRNA expression. We found that succinate levels were significantly higher in carcinoma tissues than paracancerous tissues. After succinate treatment, the colorectal cancer cell lines SW480 and HCT116 had enhanced migration and invasion, the expression of biomarkers of EMT was promoted, and significantly increased phosphorylation of STAT3. In vivo experiments also showed that succinate can increase p-STAT3 expression, promote the EMT process, and promote the distant metastasis of colorectal cancer in mice.Succinate promotes EMT through the activation of the transcription factor STAT3, thus promoting the migration and invasion of colorectal cancer.

Gastric Syphilis Mimicking Lymphoma: A Case Report.

Gastric syphilis is a rare manifestation of syphilis that occurs in about 1% of cases and is often overlooked due to its non-specific signs and symptoms. We report a case of a 28-year-old Chinese woman who presented with epigastric pain, nausea, vomiting, hematemesis, alopecia, and weight loss. She tested positive for Helicobacter pylori (H. pylori), with rapid plasma reagin (RPR) and Treponema pallidum particle assay (TPPA) tests showing titers of 1:128 and 1:320, respectively. CT imaging revealed thickening of the gastric wall, exudation around the antrum, and multiple lymphadenopathies. Gastroscopy showed multiple irregular ulcers, which resembled lymphoma. However, biopsy results did not support the presence of lymphoma, but immunohistochemistry showed an abundance of syphilis spirochetes in the mucosal lamina propria and glands. This led to a diagnosis of gastric syphilis. The patient received standard treatment for syphilis as well as anti-H. pylori therapy, and her symptoms and endoscopic findings gradually improved and eventually resolved. We hope that this case report can provide valuable insights into the diagnosis and management of gastric syphilis, which can mimic other diseases like lymphoma.

Apoprotein E methylation is correlated with immune microenvironment in hepatocellular carcinoma.

BACKGROUND: We aimed to evaluate the correlation of apoprotein E (APOE) transcription and its methylation with immune microenvironment in HCC patients. MATERIAL AND METHODS: The expression profiles of APOE transcription, APOE methylation, and APOE protein were investigated via comprehensive bioinformatic analyses. After that, the association between the immune activation of HCC and APOE transcription and methylation were analyzed. Finally, the prognostic role and immune correlation of the APOE protein in 92 HCC individuals was determined. RESULTS: Based on data from TCGA, GEO, and ICGC datasets, the APOE mRNA was differentially expressed in HCC tissues compared with normal liver tissues. Further, APOE methylation was down-regulated in HCC tissues compared to normal liver tissues. APOE methylation was negatively correlated with APOE transcription in HCC (r=-0.52, p < 0.0001). Based on APOE methylation, the HCC patients were stratified into hypermethylation and hypomethylation subgroups as they exhibited different immune activation statuses. Further, HCC individuals with APOE hypermethylation had a closer immune correlation than those with hypomethylation. Notably, APOE transcription was associated with weak immune infiltrates and activation. Finally, over-expression of the APOE protein was correlated with better survival outcomes, but not correlated with PD-1 or CTLA4 protein in HCC revealed by immunohistochemistry. CONCLUSION: APOE methylation had a closer correlation with immune cells than APOE mRNA, indicating that APOE methylation might play an important role in immune regulation in HCC.

An inflammation-associated ferroptosis signature optimizes the diagnosis, prognosis evaluation and immunotherapy options in hepatocellular carcinoma.

Inflammation and ferroptosis crosstalk complexly with immune microenvironment of hepatocellular carcinoma (HCC), thus affecting the efficacy of immunotherapy. Herein, our aim was to identify the inflammation-associated ferroptosis (IAF) biomarkers for contributing HCC. A total of 224 intersecting DEGs identified from different inflammation- and ferroptosis-subtypes were set as IAF genes. Seven of them including ADH4, APOA5, CFHR3, CXCL8, FTCD, G6PD and PON1 were used for construction of a risk model which classified HCC patients into two groups (high and low risk). HCC patients in the high-risk group exhibited shorter survival rate and higher immune score, and were predicted to have higher respond rate in immune checkpoint inhibition (ICI) therapy. Levels of the seven genes were significantly changed in HCC tissues in comparison to adjacent tissues. After inserting the gene expression into the risk model, we found that the risk model exhibited the higher diagnostic value for distinguish HCC tissues compared each single gene. Furthermore, HCC tissues from our research group with high-risk score exhibited more cases of microsatellite instability (MSI), heavier tumour mutational burden (TMB), higher expression level of PDL1 and cells with CD8. Knockdown of APOA5 reduced HCC cell proliferation combining with elevating inflammation and ferroptosis levels. In conclusion, we considered APOA5 maybe a novel target for suppressing HCC via simultaneously elevating inflammation and ferroptosis levels, and signature constructed by seven IAF genes including ADH4, APOA5, CFHR3, CXCL8, FTCD, G6PD and PON1 can act as a biomarker for optimising the diagnosis, prognosis evaluation and immunotherapy options in HCC patients.

A specific immune signature for predicting the prognosis of glioma patients with IDH1-mutation and guiding immune checkpoint blockade therapy.

Isocitrate dehydrogenase (IDH1) is frequently mutated in glioma tissues, and this mutation mediates specific tumor-promoting mechanisms in glioma cells. We aimed to identify specific immune biomarkers for IDH1-mutation (IDH1mt) glioma. The Cancer Genome Atlas (TCGA) and Chinese Glioma Genome Atlas (CGGA) were used to obtain RNA sequencing data and clinical characteristics of glioma tissues, while the stromal and immune scores of TCGA glioma tissues were determined using the ESTIMATE algorithm. Differentially expressed genes (DEGs), the protein-protein interaction(PPI) network, and least absolute shrinkage and selection operator (LASSO) and Cox regression analyses were used to select hub genes associated with stroma and immune scores and the prognoses of patients and to construct the risk model. The practicability and specificity of the risk model in both IDH1mt and IDH1-wildtype (wtIDH1) gliomas in TCGA and CGGA were evaluated. Molecular mechanisms, immunological characteristics and benefits of immune checkpoint blockade therapy in glioma tissues with IDH1mt were analyzed using GSEA, immunohistochemical staining, CIBERSORT, and T-cell dysfunction and exclusion (TIDE) analysis. The overall survival rate for IDH1mt-glioma patients with high stroma/immune scores was lower than that for those with low stroma/immune scores. A total of 222 DEGs were identified in IDH1mt glioma tissues with high stroma/immune scores. Among them, 72 genes had interactions in the PPI network, while three genes, HLA-DQA2, HOXA3, and SAA2, were selected as hub genes and used to construct risk models classifying patients into high- and low-risk score groups, followed by LASSO and Cox regression analyses. This risk model showed prognostic value in IDH1mt glioma in both TCGA and CCGA; nevertheless, the model was not suitable for wtIDH1 glioma. The risk model may act as an independent prognostic factor for IDH1mt glioma. IDH1mt glioma tissues from patients with high-risk scores showed more infiltration of M1 and CD8 T cells than those from patients with low-risk scores. Moreover, TIDE analysis showed that immune checkpoint blockade(ICB) therapy was highly beneficial for IDH1mt patients with high-risk scores. The risk model showed specific potential to predict the prognosis of IDH1mt-glioma patients, as well as guide ICB, contributing to the diagnosis and therapy of IDH1mt-glioma patients.

The Clinical Relevance and Immune Correlation of SLC10 Family Genes in Liver Cancer.

Background and Aim: This study was aimed to reveal the clinical relevance and immune correlation of the SLC10 family genes in liver cancer. Methods: A comprehensive bioinformatics analysis was utilized to determine the gene expression, genetic alterations, DNA methylation, clinical significance, survival association and immune correlation of seven SLC10 family genes in liver cancer. The multiplexed immunohistochemical technique was applied to determine the association between SLC10A3 protein expression and immune cells, and the correlation between SLC10A3 protein and immune checkpoints (PD1 and PD-L1) in a cohort of 32 individuals with liver cancer. Results: The expression of SLC10 family genes was different between normal liver tissues and malignant liver tissues. SLC10A5 showed the highest alteration rate (8%), followed by SLC10A3 (2.8%). Low expression of SLC10A1 was indicative of poor tumor grade and advanced tumor stage in liver cancer. Scatter plots uncovered that expression of SLC10A3 was inversely associated with SLC10A1 and SLC10A5 expression in liver cancer. The expression of SLC10A1 and SLC10A5 was strongly associated with their DNA methylation. SLC10A1 expression was a reliable genetic biomarker for the prediction of survival outcomes in liver cancer population. Expression of SLC10 family genes was remarkably linked with the abundance of most immune infiltrating cells in liver cancer, and SLC10A3 was the most significant member. The multiplexed immunohistochemical technique confirmed that there existed the significant correlations between SLC10A3 protein expression and CD4 T cells, CD20 B cells and the close association with PD-1 in the stromal area from malignant tissues. Conclusion: The expressions of SLC10 family genes were different between normal liver tissues and malignant liver tissues, and they were correlated with each other in liver cancer. SLC10A1 possesses the most significant correlation with survival outcomes. SLC10A3 exhibited the most significant relationship with immune cells, as revealed by bioinformatics analysis and multispectral imaging technique.

Pachymic Acid Ameliorates Pulmonary Hypertension by Regulating Nrf2-Keap1-ARE Pathway.

OBJECTIVE: Pulmonary hypertension (PH) is a severe pulmonary vascular disease that eventually leads to right ventricular failure and death. The purpose of this study was to investigate the mechanism by which pachymic acid (PA) pretreatment affects PH and pulmonary vascular remodeling in rats. METHODS: PH was induced via hypoxia exposure and administration of PA (5 mg/kg per day) in male Sprague-Dawley rats. Hemodynamic parameters were measured using a right ventricular floating catheter and pulmonary vascular morphometry was measured by hematoxylin-eosin (HE), α-SMA and Masson staining. MTT assays and EdU staining were used to detect cell proliferation, and apoptosis was analyzed by TUNEL staining. Western blotting and immunohistochemistry were used to detect the expression of proteins related to the Nrf2-Keap1-ARE pathway. RESULTS: PA significantly alleviated hypoxic PH and reversed right ventricular hypertrophy and pulmonary vascular remodeling. In addition, PA effectively inhibited proliferation and promoted apoptosis in hypoxia-induced pulmonary artery smooth muscle cells (PASMCs). Moreover, PA pretreatment inhibited the expression of peroxy-related factor (MDA) and promoted the expression of antioxidant-related factors (GSH-PX and SOD). Furthermore, hypoxia inhibited the Nrf2-Keap1-ARE signaling pathway, while PA effectively activated this pathway. Most importantly, addition of the Nrf2 inhibitor ML385 reversed the inhibitory effects of PA on ROS generation, proliferation, and apoptosis tolerance in hypoxia-induced PASMCs. CONCLUSION: Our study suggests that PA may reverse PH by regulating the Nrf2-Keap1-ARE signaling pathway.

Gustave Roussy Immune Score as a Novel Prognostic Scoring System for Colorectal Cancer Patients: A Propensity Score Matching Analysis.

AIM: The Gustave Roussy Immune Score (GRIm-Score) was originally designed to select cancer patients for immunotherapy, and later was reported to be a novel prognostic scoring system in lung cancer and esophageal cancer. This study was aimed to determine the prognostic role and predictive performance of GRIm-Score in colorectal cancer (CRC) CRC patients. METHODS: We conducted a single-institution study of 1,579 adult CRC patients receiving surgical removal, and those patients were divided into low GRIm-Score group (scores 0, 1) and high GRIm-Score group (scores 2, 3). Propensity score matching (PSM) was executed to balance the potential confounding factors between the two groups. Survival and time-dependent receiver operating characteristic (Td-ROC) analyses were applied to depict the prognostic role and predictive significance of GRIm-Score in CRC patients. RESULTS: There were 200 cases CRC patients in high GRIm-Score group and 1,379 cases in low GRIm-Score group. CRC patients with high GRIm-Score correspond with higher level of CEA, CA125, and inflammatory indexes, such as NLR, PLR, SII, PNI, and ALRI. Correlation analysis exhibited that GRIm-Score correlated well with the established inflammatory indexes. Survival analysis revealed that CRC patients in high GRIm-Score group showed worse overall survival (OS, P <0.0001) and disease-free survival (DFS, P <0.0001) compared with those in low GRIm-Score group. Results from multivariate Cox regression implicated that high GRIm-Score was not only a potent prognostic index for unfavorable OS (HR = 1.622, 95%CI: 1.118-2.355, P = 0.0109), but also a potent risk factor for worse DFS (HR = 1.743, 95%CI: 1.188-2.558, P = 0.0045). Td-ROC analysis demonstrated that GRIm-Score exhibited the superior discriminatory power in the prediction of OS and DFS when compared to SII, PNI, and ALRI. Such strong associations between high levels of preoperative GRIm-Score and unfavorable survival outcomes remained robust after PSM analysis. CONCLUSION: GRIm-Score, a novel inflammatory and nutritional risk scoring system, is a potent prognostic index in CRC patients receiving surgical removal. GRIm-Score can be used as an effective and simplified risk stratification tool for postoperative survival prediction of CRC patients.

RWR-algorithm-based dissection of microRNA-506-3p and microRNA-140-5p as radiosensitive biomarkers in colorectal cancer.

Radiotherapy resistance is one of the main causes for treatment failure in colorectal cancer (CRC), and it is associated with the deregulation of certain microRNAs. In this study, we constructed the microRNA-mRNA network consisting of 2275 microRNAs and 7045 target genes, collected the known microRNAs related to CRC-radiosensitivity (CRCR) (n=18) as the seed nodes, and applied the algorithm of random walk with restart (RWR) to the network to identify novel CRCR-related microRNAs (n=263). In functional analysis, 263 novel microRNAs shared a high proportion of the same biological processes and pathways with the known microRNAs. In topological analysis of the sub-network of the 263 microRNAs and their targets, hsa-mir-506-3p and hsa-mir-140-5p were identified as network hub nodes. In plasma, radiosensitive patients had a higher expression level of hsa-mir-506-3p and hsa-mir-140-5p than radioresistant patients. In experimental validation, both hsa-mir-506-3p and hsa-mir-140-5p over-expression could obviously decrease the cell proliferation, survival rate and colonality in CRC cells after radiation. In conclusion, this study combined the novel network-based method with experimental validation, and identified two novel radiosensitive biomarkers of hsa-mir-506-3p and hsa-mir-140-5p in CRC.

SERPINH1 regulates EMT and gastric cancer metastasis via the Wnt/ß-catenin signaling pathway.

In this study, we investigated the role of SERPINH1 in gastric cancer (GC) progression. GC patient tissues show significantly higher SERPINH1 mRNA and protein levels than normal gastric mucosal tissues. GC patients with high SERPINH1 expression are associated with lymph node metastasis and poor prognosis. SERPINH1 mRNA levels negatively correlate with E-cadherin mRNA levels and positively correlate with levels of N-cadherin, MMP2, and MMP9 mRNA levels. This suggests SERPINH1 regulates epithelial to mesenchymal transition (EMT). SERPINH1 expression was significantly higher in the HGC-27, AGS, MGC-803, and SGC-7901 GC cell lines than in the GES-1 normal gastric mucosal cell line. In SERPINH1-silenced SGC-7901 cells, survival, colony formation, migration and invasion were all reduced, whereas they were all enhanced in SERPINH1-overexpressing MGC-803 cells. Levels of WNT/ß-catenin signaling pathway proteins, including ß-catenin, Wnt2, GSK-3ß, p-GSK-3ß, NF-κB P65, Snail1, Slug and TWIST, were all reduced in SERPINH1-silenced SGC-7901 cells, and increased in the SERPINH1-overexpressing MGC-803 cells. Inhibition of SERPINH1 protein using Co1003 significantly decreased survival, invasion, and migration of GC cells. SERPINH1 thus appears to regulate EMT and GC progression via the Wnt/ß-catenin pathway, making SERPINH1 a potential prognostic biomarker and therapeutic target in GC patients.

A 4-gene prognostic signature predicting survival in hepatocellular carcinoma.

OBJECTIVE: To develop an independent prognostic signature for patients with hepatocellular carcinoma (HCC). METHODS: HCC gene expression profile the cancer genome atlas-liver hepatocellular carcinoma and GSE14520 were used as discovery and test set, respectively. Differentially expressed genes (DEGs) were identified between HCC tissues and adjacent normal liver tissues. Univariate Cox proportional hazards regression analysis was performed to identify DEGs correlated with survival of HCC patients. A 4-gene-based signature was constructed based on a least absolute shrinkage and selection operator Cox penalized regression model. The predictive value of the signature was analyzed and validated. RESULTS: Two hundred sixty-three DEGs were identified between HCC and adjacent liver tissues. After univariate survival analysis, 90 DEGs were found to be significantly correlated with the overall survival (OS) of HCC patients, of which 4 genes (KPNA2, CDC20, SPP1, and TOP2A) with non-zero coefficient were used to construct a prognostic signature. The 4-gene signature was significantly associated with the age (P = 0.046), grade ( P = 0.022), and T stage ( P = 0.023) of HCC patients in the discovery set and it also significantly associated with TNM stage ( P = 0.033), and serum alpha-fetoprotein lever ( P = 0.034). Patients in the 4-gene low-risk group were associated with better OS and recurrence-free survival (RFS) than those in the high-risk group in the discovery and test set. Meanwhile, the 4-gene signature is an independent prognostic factor regarding OS and RFS in the discovery and test set. CONCLUSION: We developed a 4-gene-based signature, which could be a candidate prognostic factor for patients with HCC.

Identification of a novel gene pairs signature in the prognosis of gastric cancer.

Current prognostic signatures need to be improved in identifying high-risk patients of gastric cancer (GC). Thus, we aimed to develop a reliable prognostic signature that could assess the prognosis risk in GC patients. Two microarray datasets of GSE662254 (n = 300, training set) and GSE15459 (n = 192, test set) were included into analysis. Prognostic genes were screened to construct prognosis-related gene pairs (PRGPs). Then, a penalized Cox proportional hazards regression model identified seven PRGPs, which constructed a prognostic signature and divided patients into high- and low-risk groups according to the signature score. High-risk patients showed a poorer prognosis than low-risk patients in both the training set (hazard ratios [HR]: 6.086, 95% confidence interval [CI]: 4.341-8.533) and test set (1.773 [1.107-2.840]). The PRGPs signature also achieved a higher predictive accuracy (concordance index [C-index]: 0.872, 95% CI: 0.846-0.897) than two existing molecular signatures (0.706 [0.667-0.744] for a 11-gene signature and 0.684 [0.642-0.726] for a 24-lncRNA signature) and TNM stage (0.764 [0.715-0.814]). In conclusion, our study identified a novel gene pairs signature in the prognosis of GC.

Regulatory role of NKG2D+ NK cells in intestinal lamina propria by secreting double-edged Th1 cytokines in ulcerative colitis.

The role of intestinal lamina propria (LP) NKG2D+ NK cells is unclear in regulating Th1/Th2 balance in ulcerative colitis (UC). In this study, we investigated the frequency of LP NKG2D+ NK cells in DSS-induced colitis model and intestinal mucosal samples of UC patients, as well as the secretion of Th1/Th2/Th17 cytokines in NK cell lines after MICA stimulation. The role of Th1 cytokines in UC was validated by bioinformatics analysis. We found that DSS-induced colitis in mice was characterized by a Th2-mediated process. In acute phrase, the frequency of LP NKG2D+ lymphocytes increased significantly and decreased in remission, while the frequency of LP NKG2D+ NK cells decreased significantly in acute phase and increased in remission. No obvious change was found in the frequency of total LP NK cells. Similarly, severe UC patients had a higher expression of mucosal NKG2D and a lower number of NKG2D+ NK cells than mild to moderate UC. In NK cell lines, the MICA stimulation could induce a predominant secretion of Th1 cytokines (TNF, IFN-γ). Furthermore, in bioinformatics analysis, mucosal Th1 cytokine of TNF, showed a double-edged role in UC when compared to the Th1-mediated disease of Crohn's colitis. In conclusion, LP NKG2D+ NK cells partially played a regulatory role in UC through secreting Th1 cytokines to regulate the Th2-predominant Th1/Th2 imbalance, despite of the concomitant pro-inflammatory effects of Th1 cytokines.

Probiotics in 14-day triple therapy for Asian pediatric patients with Helicobacter pylori infection: a network meta-analysis.

Although 14-day triple therapy has been widely administrated for eradicating Helicobacter pylori (H. pylori) in Asia, its antibiotic-associated side effects restrict the effectivity of eradication therapy in pediatric patients. Therefore, a network meta-analysis (NMA) was conducted to compare the efficacy and safety of probiotics supplemented in 14-day triple therapy in Asian pediatric patients. MATERIALS AND METHODS: Randomized controlled trials (RCTs) were retrieved comprehensively in electronic databases (such as PubMed, Cochrane library, Embase, CNKI, Wan fang database, VIP database and CBM) until April 2017. Additional references were obtained from reviewed articles. NMA was performed using a random-effects model under a frequentist framework. RESULTS: Seventeen RCTs were included. NMA indicated that Bifidobacterium infantis+Clostridium butyricum was most beneficial for H. pylori eradication rates (P-score = 0.82) and Bacillus mesentericus+Clostridium butyricum+Streptococcus faecalis for total side effects (P-score = 0.77). Taken together, Bacillus mesentericus+Clostridium butyricum+Streptococcus faecalis was the best one to supplement in 14-day triple therapy due to its efficacy (P-score = 0.72) and safety (P-score = 0.77). Additionally, pairwise meta-analysis indicated that probiotics supplemented 14-day triple therapy significantly increased H. pylori eradication rates (RR: 1.16, 95%CI: 1.07-1.26) and reduced the incidence of total side effects (RR: 0.40, 95%CI: 0.34-0.48) compared with placebo. CONCLUSIONS: Bacillus mesentericus+Clostridium butyricum+Streptococcus faecalis is the optimal probiotic regime of reducing total side effects and improving eradication rates when supplemented 14-day triple therapy. Further direct evidence is needed to warrant it.

[Long-fragment RNA inhibits hepatitis B virus gene replication and expression in HepG2.2.15 cells].

To evaluate the inhibitory effects of long antisense RNA on HBV replication in HepG2.2.15 cells. The coding region of HBV S gene was cloned into pTARGET vector in sense and antisense orientations and the recombinant plasmids were transfected into HepG2.2.15 cells which were divided into HBS2 (antisense RNA) group, HBS4 (sense RNA) group and control group. HBsAg and HBeAg in the culture supernate were detected by ELISA. The HBV DNA in the supernate was quantified by real-time PCR. After treatment, the levels of HBsAg in HepG2.2.15 cell supernatants of three groups were 0.621+/-0.027, 3.399+/-0.018 and 2.232+/-0.187 respectively; the levels of HBeAg were 0.749+/-0.019, 1.548+/-0.025 and 1.570+/-0.044 respectively and the levels of HBV DNA were 1.597+/-0.082, 3.381+/-0.297 and 3.610+/-0.063 respectively. The expressions of HBsAg and HBeAg and the HBV DNA level in HBS2 group were remarkably reduced as compared to the control (Z = -2.309, P value is less than 0.05); whereas the sense plasmid transfection (HBS4) did not affect HBeAg (Z = -0.866) and HBV DNA (Z = -1.155) levels in the culture supernate but slightly increased the HBsAg level (Z = -2.309). Antisense RNA might be a useful tool to repress HBV replication.