FMO family may serve as novel marker and potential therapeutic target for the peritoneal metastasis in gastric cancer.

Objective: To explore the relationship between flavin-containing monooxygenases (FMOs) and peritoneal metastasis (PM) in gastric cancer (GC). Materials and methods: TIMER 2.0 was used to perform pan-cancer analysis and assess the correlation between the expression of FMOs and cancers. A dataset from The Cancer Genome Atlas (TCGA) was used to analyze the correlation between FMOs and clinicopathological features of GC. PM is well established as the most common mode of metastasis in GC. To further analyze the correlation between FMOs and PM of GC, a dataset was obtained from the National Center for Biotechnology Information Gene Expression Omnibus (GEO) database. The results were validated by immunohistochemistry. The relationship between FMOs and PM of GC was explored, and a novel PM risk signature was constructed by least absolute shrinkage and selection operator (LASSO) regression analysis. The regression model's validity was tested by multisampling. A nomogram was established based on the model for predicting PM in GC patients. The mechanism of FMOs in GC patients presenting with PM was assessed by conducting Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses in TCGA and GEO datasets. Finally, the potential relationship between FMOs and immunotherapy was analyzed. Results: The pan-cancer analysis in TCGA and GEO datasets showed that FMO1 was upregulated, while FMO2 and FMO4 were downregulated in GC. Moreover, FMO1 and FMO2 correlated positively with the T and N stage of GC in the TCGA dataset. FMO1 and FMO2 expression was a risk factor for GC (hazard ratio: 1.112 and 1.185). The overexpression of FMO1 was significantly correlated with worse disease-free-survival (DFS) and overall survival (OS). However, no relationship was found between FMO2 expression in GC and DFS and OS. PM was highly prevalent among GC patients and typically associated with a worse prognosis. FMO1 was highly expressed in GC with PM. FMO1 and FMO2 were positively correlated with PM in GC. We identified a 12-gene panel for predicting the PM risk signature by LASSO (Area Under Curve (AUC) = 0.948, 95%CI: 0.896-1.000). A 10-gene panel for PM prediction was identified (AUC = 0.932, 95%CI: 0.874-0.990), comprising FMO1 and FMO2. To establish a model for clinical application, a 7-gene panel was established (AUC = 0.927, 95% CI: 0.877-0.977) and successfully validated by multisampling. (AUC = 0.892, 95% CI: 0.878-0.906). GO and KEGG analyses suggest that FMO1 and FMO2 regulate the extracellular matrix and cell adhesion. FMO1 and FMO2 were positively correlated with the immune score of GC, and their expression was associated with the infiltration of immune cells. Conclusion: PM in GC is strongly correlated with FMOs. Overall, FMO1 and FMO2 have huge prospects for application as novel diagnostic and therapeutic targets.

Circ_0001777 Affects Triple-negative Breast Cancer Progression Through the miR-95-3p/AKAP12 Axis.

BACKGROUND: Triple Negative Breast Cancer (TNBC) is 1 of the most serious cancer. Circular RNA_0001777 (circ_0001777) expression was decreased in TNBC tissues. However, the molecular mechanism of circ_0001777 remains unknown. METHODS: The expression of circ_0001777, microRNA-95-3p (miR-95-3p) and A-kinase anchor protein 12 (AKAP12) were detected by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR). A series of in vitro experiments were designed to explore the function of circ_0001777 in TNBC cells and the regulatory mechanism between circ_0001777 and miR-95-3p and AKAP12 in TNBC cells. Western blot examined the relative protein levels in TNBC cells. Bioinformatics prediction site predicted the relationship between miR-95-3p and circ_0001777 or AKAP12 and was verified by Dual-luciferase reporter assays. The xenotransplantation model was established to study the role of circ_0001777 in vivo. RESULTS: The expression of circ_0001777 and AKAP12 was decreased in TNBC tissues, while the expression of miR-95-3p was increased. Circ_0001777 can sponge miR-95-3p, and AKAP12 is the target of miR-95-3p. In vitro complement experiments, overexpression of circ_0001777 significantly decreased the malignant behavior of TNBC, while co-transfection of miR-95-3p partially up-regulated this change. In addition, AKAP12 knockdown increased the proliferation, migration, and invasion of TNBC cells inhibited by overexpression of circ_0001777. Mechanically, circ_0001777 regulates AKAP12 expression in TNBC cells by sponge miR-95-3p. In addition, in vivo studies have shown that overexpression of circ_0001777 inhibits tumor growth. CONCLUSION: Overexpression of circ_0001777 decreased proliferation, migration, and invasion of TNBC cells by regulating the miR-95-3p/AKAP12 axis, suggesting that circ_0001777/miR-95-3p/AKAP12 axis may be a potential regulatory mechanism for the treatment of TNBC.

Development of a model for predicting mortality of breast cancer admitted to Intensive Care Unit.

Background: There is still not a mortality prediction model built for breast cancer admitted to intensive care unit (ICU). Objectives: We aimed to build a prognostic model with comprehensive data achieved from eICU database. Methods: Outcome was defined as all-cause in-hospital mortality. Least absolute shrinkage and selection operator (LASSO) was conducted to select important variables which were then taken into logistic regression to build the model. Bootstrap method was then conducted for internal validation. Results: 448 patients were included in this study and 79 (17.6%) died in hospital. Only 5 items were included in the model and the area under the curve (AUC) was 0.844 (95% confidence interval [CI]: 0.804-0.884). Calibration curve and Brier score (0.111, 95% CI: 0.090-0.127) showed good calibration of the model. After internal validation, corrected AUC and Brier score were 0.834 and 0.116. Decision curve analysis (DCA) also showed effective clinical use of the model. The model can be easily assessed on website of https://breastcancer123.shinyapps.io/BreastCancerICU/. Conclusions: The model derived in this study can provide an accurate prognosis for breast cancer admitted to ICU easily, which can help better clinical management.

Survival outcomes of liver transplantation versus liver resection among patients with hepatocellular carcinoma: A SEER-based longitudinal study.

BACKGROUND/PURPOSE: This study aimed to compare the survival benefit in hepatocellular carcinoma (HCC) patients following liver transplantation or surgical resection utilizing Surveillance, Epidemiology, and End Results Program (SEER) database (2004-2013). METHODS: Overall and cancer-specific mortality were evaluated in HCC patients who were treated by liver resection or transplantation. Patients newly-diagnosed with primary HCC were included. RESULTS: Kaplan-Meier survival curves found that patients with liver transplantation had lower risk of overall mortality and cancer-free mortality than patients who received liver resection (P < 0.001). Multivariate analysis found the risk of overall and cancer-specific mortality were lower with liver transplantation than with resection (aHR = 0.51 for overall mortality and aHR = 0.37 for cancer specific mortality), and that the risk of overall mortality decreased for patients with liver transplantation relative to surgical resection as disease severity increased (T1: aHR = 0.53; T2: aHR = 0.47; T3 and T4: aHR = 0.33). CONCLUSION: The findings indicated that transplantation has survival advantages compared with resection in treating patients with HCC, particularly in later stage disease.

Pyrroloquinoline quinone protects nucleus pulposus cells from hydrogen peroxide-induced apoptosis by inhibiting the mitochondria-mediated pathway.

PURPOSE: Intervertebral disc cell apoptosis has been suggested to play a key role in promoting disc degeneration, and many studies have shown that the mechanism may be related to oxidative stress. Pyrroloquinoline quinone (PQQ), a redox cofactor for bacterial dehydrogenases, possesses the potential to scavenge reactive oxygen species (ROS) and inhibit cell apoptosis. The objective of this study was to evaluate the effects of PQQ on cultured rat nucleus pulposus (NP) cells under conditions of oxidative injury induced by hydrogen peroxide (H2O2) and to investigate the underlying mechanisms in vitro. METHODS: Cell viability was determined by CCK8 assay. Changes in the apoptosis rate, intracellular ROS levels and the mitochondrial membrane potential were measured by flow cytometry. Extracellular matrix (ECM)-related proteins (collagen-2 and aggrecan) and apoptosis-related proteins (Bcl-2, Bax, cytochrome c, and caspase-3) were investigated by western blotting. RESULTS: The results show that NP cells pretreated with PQQ before H2O2 exposure exhibited increased cell viability, decreased ROS formation, maintained mitochondrial membrane potential, and reduced apoptosis. In the presence of PQQ, ECM production was maintained by the cells despite being in an apoptotic environment. In addition, pretreatment with PQQ increased the expression of Bcl-2, inhibited the release of mitochondrial cytochrome c, and decreased the expressions of Bax and cleaved caspase-3. CONCLUSIONS: Our results suggest that PQQ can protect rat NP cells against oxidative stress via a mitochondria-mediated pathway. PQQ might be useful as a potential pharmaceutical agent in the prevention of intervertebral disc degeneration.

[Establishment of a rat model of acute liver failure by a modified 90% bloodless hepatectomy and by D-galactosamine and lipopolysaccharide injection].

OBJECTIVE: To compare the effects of different approaches to establishing rat models of acute liver failure (ALF). METHODS: Sixty-eight Sprague-Dawley rats were randomly divided into 3 groups for establishing ALF models using 3 different approaches, namely conventional hepatectomy for resecting 90% liver tissue as described by Higgins and Anderson, modified bloodless hepatectomy for resecting 90% liver tissue, and intraperitoneal injections of 700 mg/kg D-galactosamine (D-gal) and 5 µg/kg lipopolysaccharide (LPS). The mortality of the rats due to postoperative bleeding and survival rate at 7 days after the surgery were recorded. The levels of alanine aminotransferase (ALT), total bilimbin (Tbil), albumin (ALB), NH3, glucose (Glu) and prothrombin time (PT) were monitored, and histopathologies of the liver were examined at 24 and 72 h after the surgery. RESULTS: The mortality rate due to postoperative bleeding was higher in conventional hepatectomy group than in the modified surgical group (15% vs 0). The survival rate at 7 days was 25%, 0%, 15% in conventional surgical group, modified surgical group and drug injection group, respectively. In the latter two groups, significant changes of ALT, Tbil, ALB, NH3, Glu, and PT were recorded at 24 and 72 h after the modeling (P<0.05), and these changes were the most obvious at 24 h in modified surgical group and at 72 h in the drug injection group; ALB in both groups declined to the lowest at 7 days and then increased gradually. Liver cell degeneration and necrosis were found in modified surgical group and drug injection group at 24 h and 72 h after the modeling. CONCLUSION: Both the modified 90% bloodless hepatectomy and injections of D-gal and LPS can be used to establish ideal rat models of ALF to suit different ALF-related researches.

[A modified approach to establishing a rat model of liver cancer with concurrent cirrhosis].

OBJECTIVE: To establish a modified rat model of liver cancer with concurrent cirrhosis for the study of carcinogenesis characteristics and drug intervention of liver cancer. METHODS: Fifty male Wistar rats weighing 100-120 g were randomly divided into normal control group (20 rats) and model group (30 rats). In the model group, the rats were subjected to intraperitoneal injection of 50 mg/kg DEN N-diethylnitrosamine (DEN) twice a week for 4 consecutive weeks, followed then by weekly injections for another 10 weeks. The control rats received injections of 0.1 ml saline in the same manner. At 2, 4, 8, 12, 14, and 18 weeks, 3 rats from each group were sacrificed for assessing tumor formation and liver cirrhosis. RESULTS: Liver cancer with concurrent cirrhosis was induced successfully after 14 weeks of DEN injections. At the 14th week, 3 out of the 5 rats were found to have cirrhosis and LC, and at the 18th week, all the 3 rats examined had cirrhosis and liver cancer. The total carcinogenesis rate in the rats was 75% at 18 weeks with an overall mortality of 33%. CONCLUSION: This approach to establishing rat models of liver cancer with concurrent cirrhosis requires simple operation, shortens the time of carcinogenesis, and ensures a high success rate of carcinogenesis and a low mortality rate. The carcinogenesis characteristics in this model are similar to those in human.