Prevalence and Risk Factors of Metabolic Dysfunction-Associated Fatty Liver Disease with Renal Insufficiency in Overweight/Obese Adults.

INTRODUCTION: The prevalence of metabolic dysfunction-associated fatty liver disease (MAFLD) with renal insufficiency in recent years and the association between MAFLD and renal insufficiency are not entirely clear, especially in overweight/obesity. The aim of this study was to analyze the prevalence and risk factors of MAFLD with renal insufficiency in overweight/obese adults. METHODS: Individuals who attended checkup at the Second Affiliated Hospital of Xi'an Jiaotong University from 2016 to 2021 were included. The prevalence of MAFLD with renal insufficiency (estimated glomerular filtration rate ≤90 mL/min/1.73 m2) in overweight/obesity was estimated. Propensity score-matched analysis, univariate and multivariate analyses were used to determine the risk factors for MAFLD with renal insufficiency. RESULTS: From 2016 to 2021, the prevalence of MAFLD in overweight/obesity reached its highest of 44.7% in 2017 and its lowest of 36.9% in 2018; and 33.9% in 2021 and 21.8% in 2019 is the highest and lowest prevalence of MAFLD with renal insufficiency, respectively. MAFLD was more common in men, old individuals, and persons with a higher body mass index (BMI) and was characterized by significant renal insufficiency. MAFLD with renal insufficiency was more common in women, old individuals, and persons with a higher BMI and was characterized by significant metabolic dysfunction and liver fibrosis. Multivariable analysis showed that BMI, uric acid, and fibrosis (evaluated with noninvasive liver fibrosis score [fibrosis-4]) were independent risk factors for MAFLD with renal insufficiency. CONCLUSION: The prevalence of MAFLD with renal insufficiency in overweight/obese adults is quite high in the last 5 years. BMI, uric acid, and fibrosis are independent risk factors for MAFLD with renal insufficiency.

Effects of SDF-1/CXCR7 on the Migration, Invasion and Epithelial-Mesenchymal Transition of Gastric Cancer Cells.

We found that SDF-1/CXCR7 axis played an important role in the growth and proliferation of gastric cancer in the previous studies. The objectives of this study were to explore the effects of SDF-1/CXCR7 on the metastatic ability of gastric cancer cells and the possible mechanisms. CXCR7 expression in SGC-7901 gastric cancer cells was stably knocked down via lentiviral vectors. The cell migration and invasion abilities were detected by transwell migration and invasion assays. The expressions of matrix metalloproteinase 2 (MMP-2), MMP-9, vascular endothelial growth factor (VEGF), epithelial-mesenchymal transition (EMT) markers and Akt phosphorylation were detected with real-time PCR and/or western blot. We found that SDF-1 markedly enhanced the migration and invasion abilities of SGC-7901 gastric cancer cells; CXCR7 knockdown inhibited these effects. SDF-1/CXCR7 increased the expressions of MMP-2, MMP-9 and VEGF. SDF-1/CXCR7 also downregulated E-cadherin expression but upregulated N-cadherin, vimentin and Snail expressions, suggesting that SDF-1/CXCR7 could promote the development of EMT in gastric cancer cells. Furthermore, SDF-1/CXCR7 could promote Akt phosphorylation. Our results indicated that SDF-1/CXCR7 enhanced the migration, invasion and EMT of gastric cancer cells and thus CXCR7 supression may be a strategy for inhibiting gastric cancer metastasis.

Chlorogenic Acid Improves NAFLD by Regulating gut Microbiota and GLP-1.

Our previous studies have shown that chlorogenic acid (CGA) could significantly improve acute and chronic liver injury through antioxidant and anti-inflammatory activities. However, its effect on non-alcoholic fatty liver disease (NAFLD) are not entirely clear. This study aims to explore the effect of CGA on NAFLD induced by high-fat diet (HFD) and whether it regulates the gut microbiota and Glucagon-like peptide-1 (GLP-1). NAFLD mice were established by HFD and treated with or without CGA. Serum transaminase, fasting blood glucose (FBG), blood lipids, insulin, GLP-1 and lipopolysaccharide (LPS) were detected. Liver histology was evaluated with Hematoxylin-eosin staining. Toll like receptor 4 (TLR4) signaling pathway was analyzed with western blot and inflammatory cytokines were detected with real-time PCR. The content of gut microbiota were determined with real-time PCR of the bacterial 16S rRNA gene. Expressions of intestine tight junctional protein were examined with immunohistochemistry. CGA could alleviate HFD-induced hepatic steatosis and inflammation, reduce serum transaminase, FBG and blood lipids, increase insulin sensitivity. CGA also could reverse HFD-induced activation of TLR4 signaling pathway and expression of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in liver. Meanwhile, CGA increased the content of Bifidobacterium and reduced the content of Escherichia coli in feces. Furthermore, CGA could increase the expression of tight junction proteins Occludin and zonula occludens-1 (ZO-1) in intestinal tissue. Moreover, CGA could the level of LPS and increased the level of GLP-1 in portal vein. These results indicated that CGA protected against HFD-induced hepatic steatosis and inflammation probably through its anti-inflammatory effects associated with regulation of gut microbiota and an increase of GLP-1 secretion and thus could be used as a potential drug for prevention and treatment of NAFLD.

Activation of Nrf2 pathway and inhibition of NLRP3 inflammasome activation contribute to the protective effect of chlorogenic acid on acute liver injury.

Chlorogenic acid (CGA), a kind of polyphenol found in coffee, fruits and vegetables, has potent anti-oxidant and anti-inflammatory properties. Our previous studies showed CGA could efficiently alleviate liver fibrosis in rats. However, whether CGA regulates nuclear factor erythroid-2-related factor 2 (Nrf2) anti-oxidant pathway and NLRP3 inflammasome activation and protects against carbon tetrachloride (CCl4)-induced acute liver injury are unknown. We found that CGA could increase Nrf2 activation and expression of Nrf2-related anti-oxidant genes, including HO-1, NQO1 and GCLC. Pretreatment with CGA could reduce CCl4-induced elevation of serum transaminases and alleviate liver pathological abnormalities. CGA also reversed CCl4-induced increase in MDA level and decrease in the levels of GSH, SOD and CAT in liver tissues. Meanwhile, CGA inhibited NLRP3 inflammasome activation, as indicated by the reduced protein expression of NLRP3, Pro-Caspase-1, Caspase-1, Pro-IL-1ß and IL-1ß. Moreover, CGA reduced serum levels and liver mRNA expression of TNF-α, IL-6 and IL-1ß. These results demonstrate that CGA protects against CCl4-induced acute liver injury probably through enhancing Nrf2-mediated anti-oxidant pathway and inhibiting NLRP3 inflammasome activation.

Oxytocin inhibited stress induced visceral hypersensitivity, enteric glial cells activation, and release of proinflammatory cytokines in maternal separated rats.

Visceral hypersensitivity (VH) is a significant contributor to irritable bowel syndrome (IBS). Oxytocin (OT) possesses analgesic effects on the central nervous system (CNS) and attenuates microglial activation, however, little is known about its peripheral effects and involvement in VH of IBS. Reactive enteric glial cells (EGCs) contributes to abnormal motility in gastrointestinal (GI) diseases. The aim of this study was to evaluate the peripheral use of OT to maintain VH and activation of EGCs through involvement of the Toll-like receptor (TLR) 4/MyD88/NF-κB signaling. After assessing a baseline visceromotor response (VMR) to colorectal distension (CRD), rats were exposed to a 1h water avoidance stress (WAS) session. Before each WAS session, intraperitoneal injection of OT (1mg/kg body weight, in phosphate-buffered saline (PBS)) atosiban (0.5mg/kg body weight, in PBS) or PBS (as a vehicle control, 1ml/kg body weight) was administered. Animas are killed 24h after the last WAS session. EGCs activity, relative OT receptor expression, glial fibrillary acidic protein (GFAP) expression and TLR4/MyD88/NF-κB signaling were evaluated. Neonatal maternal separation (MS) significantly increased the OT receptor expression and enhanced VMR to CRD. WAS improved VMR to CRD only during neonatal MS. OT treatment prevented WAS-induced higher VMRs to CRD, which was reversed by an OT receptor antagonist administration. Compared to the vehicle, OT pre-treated rats reduced EGCs activation, GFAP expression and TLR4/MyD88/NF-κB signaling. We conclude that neonatal MS induces VH and visceral pain in rats. Furthermore, exogenous OT attenuated stress-induced VH and EGCs activation, which was mediated by TLR4/MyD88/NF-κB signaling.

Tumor-associated autoantibodies are useful biomarkers in immunodiagnosis of α-fetoprotein-negative hepatocellular carcinoma.

AIM: To determine the prevalence and diagnostic value of autoantibodies in α-fetoprotein (AFP)-negative hepatocellular carcinoma (HCC). METHODS: Fifty-six serum samples from AFP-negative HCC cases, 86 from AFP-positive HCC cases, 168 from chronic liver disease cases, and 59 from normal human controls were included in this study. Autoantibodies to nucleophosmin (NPM)1, 14-3-3zeta and mouse double minute 2 homolog (MDM2) proteins in AFP-negative HCC serum were evaluated by enzyme-linked immunosorbent assay. Partially positive sera were further evaluated by western blotting. Immunohistochemistry was used to detect the expression of three tumor-associated antigens (TAAs) in AFP-negative HCC and normal control tissues. RESULTS: The frequency of autoantibodies to the three TAAs in AFP-negative HCC sera was 21.4%, 19.6% and 19.6%, which was significantly higher than in the chronic liver disease cases and normal human controls (P < 0.01) as well as AFP-positive HCC cases. The sensitivity of the three autoantibodies for diagnosis of AFP-negative HCC ranged from 19.6% to 21.4%, and the specificity was approximately 95%. When the three autoantibodies were combined, the sensitivity reached 30.4% and the specificity reached 91.6%. CONCLUSION: Autoantibodies to NPM1, 14-3-3zeta and MDM2 may be useful biomarkers for immunodiagnosis of AFP-negative HCC.

CXCR7 as a chemokine receptor for SDF-1 promotes gastric cancer progression via MAPK pathways.

OBJECTIVE: As the alternate receptor for stromal cell-derived factor-1 (SDF-1) except CXCR4, CXCR7 has been shown to be involved in the progression of some malignancies. However, the role of SDF-1/CXCR7 in gastric cancer (GC) remains unclear. MATERIALS AND METHODS: CXCR7 expression was examined in 83 human GC tissues and adjacent non-cancer tissues (ANCTs) by immunohistochemistry, in three human GC cell lines (MGC-803, BGC-823 and SGC-7901) by reverse transcription-PCR and western blot. CXCR7 was stably knocked down via lentiviral vectors. The cells proliferation was evaluated using CCK-8 and colony formation assay; MAPK pathways (ERK1/2, p38 and SAPK/JNK) were detected using western blot. Besides, the xenograft model of nude mice for GC growth was performed. RESULTS: CXCR7 expression in GC tissues was significantly higher than that in ANCTs and associated with tumor size, TNM stage and lymph node metastasis. CXCR7 and CXCR4 were both detectable in three GC cell lines and SGC-7901 cells expressed CXCR7 the most abundantly. SDF-1 promoted the proliferation of SGC-7901 cells, the phosphorylation of ERK1/2, p38 and CXCR7 knockdown distinctly reversed these changes; the proliferation stimulated with SDF-1 was attenuated by U0126 (MEK1/2 inhibitor). Furthermore, CXCR7 knockdown inhibited the growth of GC subcutaneous xenografts and decreased the microvessel density and VEGF expression in vivo. CONCLUSION: CXCR7 was identified as a novel promoter in GC initiation and progression.

Chlorogenic acid protects against liver fibrosis in vivo and in vitro through inhibition of oxidative stress.

Liver fibrosis is a scaring process related to chronic liver injury of all causes and as yet no truly effective treatment is available. Chlorogenic acid (CGA) is a phenolic compound and exerts anti-inflammatory and anti-oxidant activities. Our former studies suggested that CGA could prevent CCl4-induced liver fibrosis through inhibition of inflammatory signaling pathway in rats. However, whether the anti-oxidant activity is involved in the anti-fibrotic effect of CGA on liver fibrosis is not yet fully understood. This study examined whether CGA may prevent CCl4-induced liver fibrosis by improving anti-oxidant capacity via activation of Nrf2 pathway and suppressing the PDGF-induced profibrotic action via inhibition of NOX/ROS/MAPK pathway. The studies in vivo showed that the liver fibrosis degree, hydroxyproline content and expression of α-SMA, Collagen Ⅰ, Collagen Ⅲ and TIMP-1 were increased in CCl4-injected rats and which were alleviated markedly by CGA. Furthermore, CGA significantly decreased CYP2E1 expression and increased the expression of nuclear Nrf2 and Nrf2-regulated anti-oxidant genes (HO-1, GCLC and NQO1). CGA decreased MDA level and increased GSH, SOD and CAT levels in liver tissues. In vitro studies PDGF could induce NOX subunits (p47phox and gp91phox) expression, ROS production, p38 and ERK1/2 phosphorylation, HSCs proliferation and profibrotic genes expression in HSCs, all of which were reduced by CGA treatment. In conclusion, the results suggest that CGA protects against CCl4-induced liver fibrosis, at least in part, through the suppression of oxidative stress in liver and hepatic stellate cells.

[Expression of CXCR7 in Gastric Cancer Cells and Its Effect on the Migration and Invasion of SGC-7901 Cells].

OBJECTIVES: To determine the expression of chemokine (C-X-C motif) receptor 7 (CXCR7) in five gastric cancer cell lines with various degrees of differentiation, and the effect of silencing CXCR7 on the migration and invasion of SGC-7901 cells. METHODS: The expression of CXCR7 in gastric cell lines (HGC-27, MGC-803, SGC-7901, BGC-823 and MKN-28) was detected by Western bolt and RT-PCR. The SGC-7901 cells were transfected with liposome of CXCR7 siRNA to silence CXCR7 gene, and then treated with stromal-derived factor-1 (SDF-1)-the ligand of CXCR7. Transwell assay was used for determining the migratory and invasive ability of SGC-7901 cells in the four groups: NC siRNA, NC siRNA+SDF-1, CXCR7 siRNA and CXCR7 siRNA+SDF-1. RESULTS: CXCR7 was expressed in the five gastric cancer cell lines, with the highest intensity in SGC-7901. The migrated and invasive cells increased in the NC siRNA+SDF-1 group and reduced in the CXCR7-siRNA group compared with the NC siRNA group (P<0.05). The CXCR7-siRNA+SDF-1 group had less migrated and invasive cells than the NC siRNA+SDF-1 group (P<0.05). CONCLUSIONS: CXCR7 is highly expressed in SGC-7901. SDF-1 promotes the migratory and invasive capability of SGC-7901 cells, but such an effect can be inhibited by silencing it with CXCR7siRNA.

[Effect of Chlorogenic Acid on Hepatic Stellate Cell Proliferation, Generation and Degradation of Extracelluar Matrix].

OBJECTIVE: To investigate the effects of chlorogenic acid (CGA) on hepatic stellate cell proliferation and the expression and secretion of Collagen I, Collagen III, tissue inhibitors of metalloproteinase-1 (TIMP-1) and matrix metalloproteinase-2 (MMP-2). METHODS: An immortalized rat hepatic stellate cell (HSC) line was cultured in vitro. The cells were divided into 5 groups: control group; platelet-derived growth factor (PDGF) (10 ng/mL), PDGF+CGA (12.5 µg/mL), PDGF+CGA (25 µg/mL), PDGF+CGA (50 µg/mL) and CGA (50 µg/mL) group. After 24 hours treatment, the proliferation of HSC was detected by MTT method. The mRNA expression of Collagen I, Collagen III, TIMP-1 and MMP-2 were detected by RT-PCR. The protein levels of Collagen I, Collagen III, TIMP-1 and MMP-2 in the culture supernatant of HSC were measured by ELISA. RESULTS: PDGF increased the hepatic stellate cell proliferation, the mRNA expression and the protein levels of Collagen I, Collagen III and TIMP-1. (P < 0.05), which were significantly decreased by CGA (P < 0. 05). However, CGA had no significant influence on the expression of MMP-2. CONCLUSION: The antifibrotic effect of CGA may be related with the inhibition of hepatic stellate cell proliferation and generation of extracelluar matrix and promotion of extracelluar matrix degradation.

Association between the expression of T-cadherin and vascular endothelial growth factor and the prognosis of patients with gastric cancer.

T-cadherin has been identified as a tumor-suppressor gene in several types of cancer. The present study aimed to investigate the association of the expression of T-cadherin with angiogenesis, and to evaluate its prognostic value for patients with primary gastric cancer. Gastric cancer tissues and matched adjacent tissues from 166 patients receiving surgical resection were included in the present study. The expression of T-cadherin was detected using immunohistochemistry, western blotting and reverse transcription-quantitative polymerase chain reaction. The expression of vascular epidermal growth factor (VEGF) was detected using immunohistochemistry, and its association with the expression of T-cadherin was analyzed. In addition, the association between the expression of T-cadherin and clinicopathological features were analyzed. The mRNA and protein expression levels of T-cadherin were significantly lower in the gastric cancer tissue compared with the corresponding adjacent normal tissue (P<0.05). The expression of VEGF was not associated with the expression of T-cadherin in the gastric cancer tissue. The decreased protein expression of T-cadherin correlated with smoking, larger tumor size (diameter, >4 cm), lymph node metastasis and a higher tumor-lymph node-metastasis stage (P<0.05 or P<0.01). However, the expression of T-cadherin was not correlated with gender, age, alcohol intake, Helecobacter pylori infection or differentiation (P>0.05). The multivariate analysis demonstrated that the expression of T-cadherin was an independent prognostic factor for the overall survival rate of patients with gastric cancer. This data suggested that the downregulation of T-cadherin may contribute to gastric cancer progression, representing a useful biomarker for predicting the biological behavior and prognosis of gastric cancer. However, no significant association was observed between the expression of VEGF and T-cadherin.

[Expression of chemokine receptor CXCR7 in gastric cancer tissues and cell lines].

OBJECTIVE: To investigate the expression status of CXCR7 in gastric cancer tissues and cell lines. METHODS: The expression status of CXCR7 was detected in 35 primary gastric cancer tissues and matched adjacent tissues by immunohistochemistry and RT-PCR. The correlation of CXCR7 expression with the clinicopathological parameters and risk factors of gastric cancer was analyzed. The expression of CXCR7 in gastric cell lines (HGC-27, MGC-803, BGC-823, SGC-7901 and MKN-28) was also detected by immunofluorescence assay. RESULTS: The expression of CXCR7 was significantly higher in gastric cancer tissues than in adjacent tissues (P<0.01). CXCR7 expression was not correlated with age, gender, smoking history, Helicobacter pylori infection, tumor location or the pathological type, but showed a higher expression level in patients with a alcohol-drinking history than in those without (P<0.05). CXCR7 was expressed with variable intensities in the 5 gastric cancer cell lines without correlation with the degrees of cell differentiation; its expression was the highest in SGC-7901 cells, a moderately differentiated human gastric adenocarcinoma cell line. CONCLUSIONS: CXCR7 is highly expressed in gastric cancer tissues with variable intensities in 5 gastric cancer cell lines, suggesting its important role in gastric cancer progression.

Decreased expression of gastrokine 1 in gastric mucosa of gastric cancer patients.

AIM: To investigate the expression of gastrokine 1 (GKN1) in normal gastric mucosa, precancerous lesions and gastric cancer tissues, and to analyse its correlations with tumour site and pathological pattern. METHODS: Thirty gastric cancer patients (12 cases of diffuse type and 18 cases of intestinal type), 13 atrophic gastritis patients and 15 healthy volunteers with almost normal gastric mucosa (superficial gastritis) were enrolled in this study. Helicobacter pylori (H. pylori) infection was examined in all subjects. All gastric mucosa biopsy specimens were obtained. Cancer-adjacent specimens were taken from corresponding gastric cancer patients. Immunohistochemistry and real-time PCR were performed to determine the expressions of the GKN1 protein and mRNA, respectively. RESULTS: H. pylori infection had no significant association with age, gender, tumour site or pathological pattern in all subjects. Compared with the superficial gastritis and atrophic gastritis groups, the expression of GKN1 protein (P = 0.011) and mRNA (P < 0.001) in gastric cancer was significantly decreased. The GKN1 mRNA level in diffuse type gastric cancer was significantly lower than in intestinal type gastric cancer (0.296 ± 0.076 vs 0.525 ± 0.164, P < 0.001). CONCLUSION: Compared with almost normal gastric mucosa, GKN1 expression in the gastric mucosa of gastric cancer patients is decreased; this is associated with progression and prognosis of gastric cancer.