Prevalence of and associated factors for sarcopenia in patients with liver cirrhosis: A systematic review and meta-analysis.

BACKGROUND: Sarcopenia is associated with poor outcomes in patients with cirrhosis. However, the prevalence of and associated factors for developing sarcopenia in this population remain to be determined. OBJECTIVES: This study aimed to summarize the prevalence, characteristics, and associated factors of sarcopenia in patients with cirrhosis. METHODS: Electronic searches were performed from inception to June 9, 2022 to identify the eligible studies. We meta-analyzed the prevalence of sarcopenia in overall patients with cirrhosis and subgroups. Both crude and adjusted odds ratios (ORs) were pooled using the random effects model. RESULTS: A total of 55 studies involving 13,158 patients from 17 countries were included. The overall prevalence of sarcopenia was 40.1 % (95 % CI 35.4%-44.9 %) in patients with cirrhosis. The pooled prevalence was higher in males, Child-Pugh class C cirrhosis, decompensated stage, ascites, subjective global assessment class C cirrhosis, and when sarcopenia was defined by L3-SMI (third lumbar-skeletal muscle index) at a higher cutoff. In multivariate analysis, older age (adjusted OR 1.04, 95 % CI 1.00-1.07), male (adjusted OR 4.75, 95 % CI 2.72-8.28), lower body mass index (BMI) (adjusted OR 0.78, 95 % CI 0.73-0.83), alcoholic liver disease (ALD) (adjusted OR 1.43, 95 % CI 1.19-1.72), but not ascites and hepatic encephalopathy, were significantly associated with an increased risk of sarcopenia in patients with cirrhosis. CONCLUSION: Sarcopenia is a prevalent complication, and older age, male patients, lower BMI, and patients with ALD are associated with an increased risk of sarcopenia in patients with cirrhosis.

Cuproptosis regulatory genes greatly contribute to clinical assessments of hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is a common abdominal cancer with dissatisfactory therapeutic effects. The discovery of cuproptosis lights on new approach for cancer treatment and assessment. So far, there is extremely limited research investigating the roles of cuproptosis-related (CR) genes in cancers. METHODS: A novel CR risk signature was constructed using the Lasso regression analysis. Its prognostic value was assessed via a series of survival analyses and validated in three GEO cohorts. The effects of CR risk signature on tumor immune microenvironment (TIM) were explored through CIBERSORT, ESTIMATE, and ssGSEA algorithms. Using GESA, we investigated its impacts on various metabolism process. The somatic mutation features of CR signature genes were also explored via cBioPortal database. Using tumor mutation burden, expressions of immune checkpoints, TIDE score, IMvigor 210 cohort, and GSE109211 dataset, we explored the potential associations of CR risk score with the efficacy of immune checkpoint inhibitors (ICIs) and sorafenib. Finally, the biofunctions of DLAT in HCC cells were ascertained through qPCR, immunohistochemistry, colony formation, and Transwell assays. RESULTS: FDX1, DLAT, CDKN2A and GLS constituted the CR risk signature. CR risk signature possessed high prognostic value and was also applicable to three validation cohorts. Meanwhile, it could improve the accuracy and clinical making-decision benefit of traditional prognostic model. Moreover, high CR risk was indicative of unfavorable anti-tumor immune response and active metabolisms of glycolysis and nucleotide. As for therapeutic correlation, CR risk score was a potential biomarker for predicting the efficacy of ICIs and sorafenib. Through qPCR and immunohistochemistry detection in clinical samples, we reconfirmed DLAT was significantly upregulated in HCC samples. Overexpression of DLAT could promote the proliferation, migration, and invasion of HepG2 and HuH-7 cells. CONCLUSIONS: The novel CR risk signature greatly contributed to the clinical assessment of HCC. Cuproptosis regulatory gene DLAT possessed cancer-promoting capacities and was expected to be a promising therapeutic target for HCC.

Suppression of rhomboid domain-containing 1 produces anticancer effects in pancreatic adenocarcinoma through affection of the AKT/GSK-3ß/ß-catenin pathway.

The protumor role of rhomboid domain-containing 1 (RHBDD1) has been observed in multiple cancers. However, the relationship between RHBDD1 and pancreatic adenocarcinoma has not been addressed. This project focused on the potential relevance of RHBDD1 in pancreatic adenocarcinoma. Bioinformatic analysis by publicly available data revealed that RHBDD1 was abundantly expressed in pancreatic adenocarcinoma. We further verified that RHBDD1 was expressed highly in clinical specimens of pancreatic adenocarcinoma. The Kaplan-Meier curve demonstrated that high-RHBDD1 expression was associated with poor prognosis in pancreatic adenocarcinoma patients. The functional studies revealed that depletion of RHBDD1 produced in vitro anticancer effects in pancreatic adenocarcinoma cells, including retardation of proliferation, reduction of metastatic potential, and induction of cell-cycle arrest at the G0/G1 phase and apoptosis. Mechanistic studies indicated that loss of RHBDD1 affected the activation of ß-catenin via regulation of AKT. Forced expression of ß-catenin reversed the RHBDD1-loss-induced anticancer effects in pancreatic adenocarcinoma cells. Crucially, depletion of RHBDD1 retarded the growth of pancreatic adenocarcinoma xenografts in vivo, a phenomenon associated with the AKT/ß-catenin pathway. Collectively, these findings delineated that restraint of RHBDD1 displayed remarkable anticancer effects in pancreatic adenocarcinoma by affecting the AKT/ß-catenin pathway. Our work unveils a pivotal role of RHBDD1 in pancreatic adenocarcinoma and proposes it as a novel candidate target for anticancer therapy of pancreatic adenocarcinoma.

Long noncoding RNA RP11-909N17.2 promotes proliferation, invasion, and migration of hepatocellular carcinoma by regulating microRNA-767-3p.

Hepatocellular carcinoma (HCC) is one of the most common causes of cancer-related deaths worldwide, especially in developing countries. Although advances in surgical procedures and targeted medicine have improved the overall survival of patients with HCC, the prognosis is poor. Hence, there is a need to identify novel therapeutic targets for HCC. Here, we report that the expression of RP11-909N17.2, a novel, long, noncoding RNA (lncRNA), is dysregulated in patients with HCC and cell lines. Additionally, this study demonstrated that RP11-909N17.2 facilitates the proliferation and invasion of HCC cells by binding to miRNA-767-3p, a tumor-suppressive microRNA (miRNA). Small integral membrane protein 7 (SMIM7) was identified as the downstream target of miRNA-767-3p. The expression of SMIM7 was upregulated in HCC clinical samples and cell lines. Moreover, SMIM7 was involved in the proliferation and invasion of HCC cells. Furthermore, SMIM7 inhibited the apoptosis of HCC cells, which indicated the oncogenic role of SMIM7 in HCC. The findings of this study suggest that the lncRNA-miRNA-mRNA regulatory axis, which regulates the pathogenesis of HCC, can be a potential novel diagnostic and therapeutic target for HCC.

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.

Schizandrin B inhibits the cis­DDP­induced apoptosis of HK­2 cells by activating ERK/NF­κB signaling to regulate the expression of survivin.

The nephrotoxicity of cisplatin limits its clinical application. Schizandrin B (SchB) has been demonstrated to have a variety of potential cytoprotective activities. The present study explored the molecular mechanisms by which SchB inhibits the dichlorodiammine platinum (DDP)­induced apoptosis of HK­2 proximal tubule epithelial cells. In vitro assays demonstrated that SchB increased the viability of HK­2 cells, alleviated the cis­DDP­induced activation of caspase­3, reduced apoptosis and improved the nuclear morphology of HK­2 cells. Additionally, the mechanism underlying the cis­DDP­induced apoptosis was indicated to involve the activation of p53, c­Jun­N­terminal kinase (JNK) and p38 signaling. Furthermore, SchB was demonstrated to activate extracellular signal­regulated kinase (ERK) and nuclear factor κB (NF­κB) signaling, and induce the expression of survivin. The inhibition of ERK and NF­κB signaling using U0126 and pyrollidine dithiocarbamate, respectively, inhibited the expression of survivin, whereas blocking the expression of survivin using small interfering RNA inhibited the alleviating effect of SchB on cis­DDP­induced apoptosis as indicated by a reduction in cleaved caspase­3 expression. In conclusion, SchB regulates ERK/NF­κB signaling to induce the expression of survivin, thereby alleviating cis­DDP­induced renal injury.

Paeoniflorin ameliorates renal function in cyclophosphamide-induced mice via AMPK suppressed inflammation and apoptosis.

The study is to investigate the effects of paeoniflorin (PA) on renal function in cyclophosphamide-induced mice. Mice were injected with intraperitoneal cyclophosphamide (CYP, 200mg/kg) or saline respectively. Mice were treated with PA (15, 30mg/kg/day) or vehicle for the next 7 days. Then, mice were sacrificed to analyze the biochemical, histological parameters and mechanism research. Our results shown that PA significantly decreased the urine levels of uric acid and creatinine, serum and kidney levels of cytokines such as interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α), PA also obviously attenuated the histological changes of the kidney tissues caused by CYP. Moreover, Western blot demonstrated that PA increased the AMPK levels and inhibited NF-κB signaling pathway and apoptosis in CYP-stimulated kidney tissues. In conclusion, PA might be considered as an effective agent in the amelioration of the kidney toxicity resulting from CYP treatment.

Activation of the NRF2-ARE signalling pathway by the Lentinula edodes polysaccharose LNT alleviates ROS-mediated cisplatin nephrotoxicity.

The nephrotoxicity of cisplatin (cis-DDP) limits its general clinical applications. Lentinan (LNT), a dextran extracted from the mushroom Lentinula edodes, has been shown to have multiple pharmacological activities. The primary objective of the current study was to determine whether and how LNT alleviates cis-DDP- induced cytotoxicity in HK-2 cells and nephrotoxicity in mice. LNT did not interfere with cisplatin's anti-tumour efficacy in vitro and functioned cooperatively with cis-DDP to inhibit activity in HeLa and A549 tumour cells. LNT alleviated the cis-DDP-induced decrease in HK-2 cell viability, caspase-3 activation and cleavage of the DNA repair enzyme PARP, decreased HK-2 cell apoptosis and inhibited reactive oxygen species (ROS) accumulation in HK-2 cells. The inhibitor of ROS (N-acetyl-L-cysteine, NAC) could decreased the apoptosis of HK-2 cell. In addition, LNT significantly prevented cis-DDP-induced kidney injury in vivo. LNT itself could not eliminate ROS levels in vitro. Further studies demonstrated that LNT induced NF-E2 p45-related factor 2 (Nrf2) protein and mRNA expression in a time- and dose-dependent manner. LNT promoted Nrf2 translocation to the nucleus and binding to the antioxidant-response element (ARE) sequence and induced the transcription and translation of heme oxygenase 1 (HO-1), aldo-keto reductases 1C1 and 1C2 (AKR1C), and NADP(H):quinone oxidoreductase 1 (NQO1). Finally, we used hNrf2 siRNA and an Nrf2 agonist (tBHQ) to inhibit or enhance Nrf2 expression. The results demonstrated that the LNT-mediated alleviation of cis-DDP-induced nephrotoxicity was achieved by preventing the accumulation of ROS in a manner that depended on the activation of the Nrf2-ARE signalling pathway.

Lentinan mitigates therarubicin-induced myelosuppression by activating bone marrow-derived macrophages in an MAPK/NF-κB-dependent manner.

Bone marrow (BM) suppression (also known as myelosuppression) is the most common and most severe side-effect of therarubicin (THP) and thereby limits the clinical application of this anticancer agent. Lentinan (LNT), a glucan extracted from dried shiitake mushrooms (Lentinula edodes), exhibits a variety of pharmacological activities. The objectives of the present study were to determine the effect of LNT on the myelosuppression of THP-treated mice and to examine the pharmacological mechanism of these effects. In vivo experiments indicated that non-cytotoxic levels of LNT strongly increased blood myeloperoxidase (MPO) activity; improved BM structural injuries; increased the numbers of leukocytes and neutrophils in the blood and BM; elevated the blood levels of granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF); and reduced the self-healing period in THP-treated mice. In vitro experiments indicated that LNT increased the viability of BM-derived macrophages (BMDMs) in a time- and dose-dependent manner without toxic side-effects and markedly increased the release of G-CSF, GM-CSF and M-CSF by BMDMs. Further analyses revealed that LNT activated the NF-κB and MAPK signalling pathways and promoted the nuclear import of p65 and that BAY 11-7082 (a specific inhibitor of NF-κB) suppressed the release of G-CSF, GM-CSF and M-CSF. Furthermore, we found that U0126, SB203580 and SP600125 (specific inhibitors of ERK, p38 and JNK, respectively) markedly inhibited the IKK/IκB/NF-κB-dependent release of G-CSF, GM-CSF and M-CSF. In conclusion, LNT induces the production of G-CSF, GM-CSF and M-CSF by activating the MAPK/NF-κB signalling pathway in BM cells, thereby mitigating THP-induced myelosuppression.

[Changes of left ventricular function in cirrhotic patients and their correlation with the model for end-stage liver disease score].

OBJECTIVE: To investigate the changes of left ventricular structure and function in patients with liver cirrhosis and their correlation with the model for end-stage liver disease (MELD) score. METHODS: A total of 89 cirrhotic patients admitted between June, 2012 and June, 2014 and 30 healthy control subjects were enrolled in the study. According to MELD score, the cirrhotic patients were divided into 3 groups with MELD scores ≤9, between 10 and 19, and ≥20. The parameters of the left ventricle in resting state were measured using Doppler echocardiography, including left ventricular end systolic diameter (LVESD), left ventricular end diastolic diameter (LVEDD), interventricular septal thickness (IVST), left ventricular posterior wall thickness (LVPWT), left atrial diameter (LAD), ejection fraction (LVEF), cardiac output (CO), mitral flow velocity, and E wave deceleration time (DT), and evaluated their relationship with MELD score. RESULTS: Compared with the control subjects, the cirrhotic patients showed significantly increased LVESD, LVEDD, IVST, LAD, CO and DT but reduced VE/VA ratio (P<0.05 or 0.01). The values of LVESD, LVEDD, IVST, LAD and DT increased gradually with MELD scores (P<0.05 or 0.01). VE/VA ratio was higher in patients with MELD score of 10-19 than in those with MELD score ≤9, and decreased significantly in those with MELD score ≥20. Of the cirrhotic patients, 55% were found to have left atrial enlargement and 44% had a VE/VA ratio ≤1; left atrial enlargement and a VE/VA ratio below 1 were more common in patients with a MELD score ≥20 than in those with lower MELD scores. The LAD, LVEDD and DT were positively correlated with MELD scores (r=0.208, 0.319 and 0.197, respectively; P<0.05 or 0.01). CONCLUSIONS: The patients with liver cirrhosis can have cardiac function deficiency manifested mainly by left ventricular diastolic dysfunction in positive correlation with the severity of liver disease.

Silencing of NUF2 inhibits tumor growth and induces apoptosis in human hepatocellular carcinomas.

BACKGROUND: As an important component of the NDC80 kinetochore complex, NUF2 is essential for kinetochore-microtubule attachment and chromosome segregation. Previous studies also suggested its involvement in development of various kinds of human cancers, however, its expression and functions in human hepatocellular carcinoma (HCC) are still unclear. MATERIALS AND METHODS: In the present study, we aimed to test the hypothesis that NUF2 is aberrant in human HCCs and associated with cell growth. RESULTS: Our results showed significantly elevated expression of NUF2 in human HCC tissues compared to adjacent normal tissues, and high expression of NUF2 in HCC cell lines. Using lentivirus-mediated silencing of NUF2 in HepG2 human HCC cells, we found that NUF2 depletion markedly suppressed proliferation and colony formation capacity in vitro, and dramatically hampered tumor growth of xenografts in vivo. Moreover, NUF2 silencing could induce cell cycle arrest and trigger cell apoptosis. Additionally, altered levels of cell cycle and apoptosis related proteins including cyclin B1, Cdc25A, Cdc2, Bad and Bax were also observed. CONCLUSIONS: In conclusion, these results demonstrate that NUF2 plays a critical role in the regulation of HCC cell proliferation and apoptosis, indicating that NUF2 may serve as a potential molecular target for therapeutic approaches.

Cirrhosis-related changes in left ventricular function and correlation with the model for end-stage liver disease score.

OBJECTIVE: The purpose of our study is to investigate cirrhosis-related left ventricular remodeling and functional changes, further to analyze the correlations with model for end-stage liver disease (MELD) score. METHODS: A total of 89 cirrhotic patients were enrolled for study and subgrouped according to MELD score: ≤ 9, 10-19, and ≥ 20. Thirty healthy individuals were enrolled as controls. All study participants underwent cardiac assessment of the left ventricle with Doppler echocardiography; the parameters assessed included left ventricular-end systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD), interventricular septal thickness (IVST), left ventricular posterior wall thickness (LVPWT), left atrial diameter (LAD), left ventricular ejection fraction (LVEF), cardiac output (CO), mitral flow velocity (VE/VA ratio), and E-wave deceleration time (DT). RESULTS: The cirrhotic patients had significantly higher LVESD, LVEDD, IVST, LAD, CO and DT than the control group, but significantly lower VE/VA ratio (all P < 0.05). Subgroup analysis showed that the higher the MELD score, the greater the increase in LVESD, LVEDD, IVST, LAD and DT (all P < 0.05). Nearly one-half of the cirrhotic patients showed left atrial enlargement and a VE/VA ratio ≤ 1, and these features were more common in patients with MELD score ≥ 20. LAD, LVEDD and DT were positively correlated with MELD score (r = 0.208, 0.319 and 0.197, respectively; all P < 0.05). CONCLUSIONS: Patients with cirrhosis had impaired cardiac function, mainly present as left ventricular diastolic dysfunction, and the extent of dysfunction was correlated with the MELD score. Left atrial enlargement and VE/VA ratio ≤ 1 may serve as useful diagnostic indexes for cirrhotic cardiomyopathy.