Cancer-associated fibroblasts induce sorafenib resistance of hepatocellular carcinoma cells through CXCL12/FOLR1.

BACKGROUND: Due to the high drug resistance of hepatocellular carcinoma (HCC), sorafenib has limited efficacy in the treatment of advanced HCC. Cancer-associated fibroblasts (CAFs) play an important regulatory role in the induction of chemoresistance. This study aimed to clarify the mechanism underlying CAF-mediated resistance to sorafenib in HCC. METHODS: Immunohistochemistry and immunofluorescence showed that the activation of CAFs was enhanced in HCC tissues. CAFs and paracancerous normal fibroblasts (NFs) were isolated from the cancer and paracancerous tissues of HCC, respectively. Cell cloning assays, ELISAs, and flow cytometry were used to detect whether CAFs induced sorafenib resistance in HCC cells via CXCL12. Western blotting and qPCR showed that CXCL12 induces sorafenib resistance in HCC cells by upregulating FOLR1. We investigated whether FOLR1 was the target molecule of CAFs regulating sorafenib resistance in HCC cells by querying gene expression data for human HCC specimens from the GEO database. RESULTS: High levels of activated CAFs were present in HCC tissues but not in paracancerous tissues. CAFs decreased the sensitivity of HCC cells to sorafenib. We found that CAFs secrete CXCL12, which upregulates FOLR1 in HCC cells to induce sorafenib resistance. CONCLUSIONS: CAFs induce sorafenib resistance in HCC cells through CXCL12/FOLR1.

Quantitative N-glycoproteomic analysis of egg yolk powder during thermal processing.

The production of egg yolk powder often involves critical processes such as pasteurization and spray drying, however, these thermal processes will inevitably affect the functional properties of egg yolk (especially gelation and emulsification). The aim of this study was to elucidate the mechanism of the effect of pasteurized egg yolk (P-EY) and spray-dried egg yolk (SD-EY) on the functional properties through quantitative N-glycoproteomic. The results showed, compared with fresh egg yolk (F-EY), emulsifying property of mild heat-treated P-EY was slightly reduced while the gelation property did not undergo significant changes, whereas emulsifying activity (EAI) and gelation strength of vigorously heat-treated SD-EY were significantly reduced by 48.72 % and 35.73 %, respectively. During thermal processing in SD-EY, larger aggregate particles (particle size ∼10 um) were formed, and the surface hydrophobicity was reduced (93.0 %) and the zeta potential was enhanced (62.8 %). The results of quantitative N-glycoproteomic showed that 13 N-glycosylated proteins (APOB, vitellogenin, etc.) were down-regulated while only 2 N-glycosylated proteins were up-regulated; 21 N-glycosylation sites were down-regulated and 2 N-glycosylation sites were up-regulated in SD-EY, suggesting that covalent cross-linking of protein N-glycoproteins may have occurred in the process of spray-drying, which altered molecular physicochemical characteristics of the yolk solution that further affecting the processing properties of egg yolk.

Uncovering the Carboxylated Metabolome in Gut Microbiota-Host Co-metabolism: A Chemical Derivatization-Molecular Networking Approach.

Gut microbiota-host co-metabolites serve as essential mediators of communication between the host and gut microbiota. They provide nutrient sources for host cells and regulate gut microenvironment, which are associated with a variety of diseases. Analysis of gut microbiota-host co-metabolites is of great significance to explore the host-gut microbiota interaction. In this study, we integrated chemical derivatization, liquid chromatography-mass spectrometry, and molecular networking (MN) to establish a novel CD-MN strategy for the analysis of carboxylated metabolites in gut microbial-host co-metabolism. Using this strategy, 261 carboxylated metabolites from mouse feces were detected, which grouped to various classes including fatty acids, bile acids, N-acyl amino acids, benzoheterocyclic acids, aromatic acids, and other unknown small-scale molecular clusters in MN. Based on the interpretation of the bile acid cluster, a novel type of phenylacetylated conjugates of host bile acids was identified, which were mediated by gut microbiota and exhibited a strong binding ability to Farnesoid X receptor and Takeda G protein-coupled receptor 5. Our proposed strategy offers a promising platform for uncovering carboxylated metabolites in gut microbial-host co-metabolism.

Quantitative lipidomics analysis of changes in egg yolk lipids during spray-drying and subsequent accelerated storage.

Egg yolks are rich in lipids that are easily altered during processing and storage. In this study, a liquid chromatography-tandem mass spectrometry strategy was used for quantitative lipidomics analysis of egg yolk after spray-drying processing and accelerated storage. Spray-drying treatment caused lipid oxidation (especially the oxidation of free fatty acids), potential hydrolysis of phospholipids, and alteration of the form of certain polyunsaturated fatty acids (docosahexaenoic acid, eicosapentaenoic acid, linolenic acid, and eicosatetraenoic acid) in egg yolk. These lipid alterations caused by the spray-drying process were further aggravated by the accelerated storage process. In detail, following storage, the abundance of free fatty acids, phosphatidic acid and phosphatidylethanolamine decreased further; and the abundance of polyunsaturated fatty acids in the form of triglycerides increased significantly. These results provide new insight into the mechanism underlying egg yolk property changes during spray-drying and storage, and offer valuable reference data for egg yolk powder promotion and application in food processing.

Self-Training for Class-Incremental Semantic Segmentation.

In class-incremental semantic segmentation, we have no access to the labeled data of previous tasks. Therefore, when incrementally learning new classes, deep neural networks suffer from catastrophic forgetting of previously learned knowledge. To address this problem, we propose to apply a self-training approach that leverages unlabeled data, which is used for rehearsal of previous knowledge. Specifically, we first learn a temporary model for the current task, and then, pseudo labels for the unlabeled data are computed by fusing information from the old model of the previous task and the current temporary model. In addition, conflict reduction is proposed to resolve the conflicts of pseudo labels generated from both the old and temporary models. We show that maximizing self-entropy can further improve results by smoothing the overconfident predictions. Interestingly, in the experiments, we show that the auxiliary data can be different from the training data and that even general-purpose, but diverse auxiliary data can lead to large performance gains. The experiments demonstrate the state-of-the-art results: obtaining a relative gain of up to 114% on Pascal-VOC 2012 and 8.5% on the more challenging ADE20K compared to previous state-of-the-art methods.

Class-Incremental Learning: Survey and Performance Evaluation on Image Classification.

For future learning systems, incremental learning is desirable because it allows for: efficient resource usage by eliminating the need to retrain from scratch at the arrival of new data; reduced memory usage by preventing or limiting the amount of data required to be stored - also important when privacy limitations are imposed; and learning that more closely resembles human learning. The main challenge for incremental learning is catastrophic forgetting, which refers to the precipitous drop in performance on previously learned tasks after learning a new one. Incremental learning of deep neural networks has seen explosive growth in recent years. Initial work focused on task-incremental learning, where a task-ID is provided at inference time. Recently, we have seen a shift towards class-incremental learning where the learner must discriminate at inference time between all classes seen in previous tasks without recourse to a task-ID. In this paper, we provide a complete survey of existing class-incremental learning methods for image classification, and in particular, we perform an extensive experimental evaluation on thirteen class-incremental methods. We consider several new experimental scenarios, including a comparison of class-incremental methods on multiple large-scale image classification datasets, an investigation into small and large domain shifts, and a comparison of various network architectures.

Efficacy of Near-Infrared Fluorescence-Guided Hepatectomy for the Detection of Colorectal Liver Metastases: A Randomized Controlled Trial.

BACKGROUND: The application of indocyanine green fluorescence-guided hepatectomy for liver metastases from colorectal cancer is in the preliminary stage of clinical practice; thus, its efficacy needs to be determined. This study compared the number of intrahepatic colorectal liver metastases detected intraoperatively and postoperative recovery data between patients who underwent traditional hepatectomy (nonindocyanine green group) and traditional hepatectomy plus intraoperative indocyanine green fluorescence imaging (indocyanine green group). STUDY DESIGN: Between January 2018 and March 2020, patients with potentially resectable colorectal liver metastases were randomly assigned to the nonindocyanine green or indocyanine green group. The number of intrahepatic colorectal liver metastases identified intraoperatively and based on postoperative recovery data were compared between both groups. RESULTS: Overall, we recruited 80 patients, among whom 72 eligible patients were randomly assigned. After allocation, 64 patients, comprising 32 in each group, underwent the allocated intervention and follow-up. Compared with the nonindocyanine green group, the mean number of intrahepatic colorectal liver metastases identified intraoperatively in the indocyanine green group was significantly greater (mean [standard deviation], 3.03 [1.58] vs 2.28 [1.35]; p = 0.045), the postoperative hospital stay was shorter (p = 0.012) and the 1-year recurrence rate was also lower (p = 0.017). Postoperative complications and 90-day mortality were comparable, with no statistical differences. CONCLUSIONS: Indocyanine green fluorescence imaging significantly increases the number of intrahepatic colorectal liver metastases identified and reduces postoperative hospital stay and 1-year recurrence rate without increasing hepatectomy-related complications and mortality rates.

Efficacy of Adjuvant Transarterial Chemoembolization after Radical Hepatectomy in Solitary Hepatocellular Carcinoma Patients: A Retrospective Study.

Background: More and more studies have suggested that hepatocellular carcinoma (HCC) patients with high-risk recurrence factors can benefit the most from postoperative adjuvant transarterial chemoembolization (PA-TACE) for its potential effect in delaying cancer recurrence. However, it remains unclear if solitary HCC (SHCC) patients particularly those without high-risk recurrence factors should also receive PA-TACE. This study aimed to analyze the efficacy of PA-TACE in them. Methods: Retrospectively, we enrolled 123 SHCC patients who either received radical hepatectomy alone (No TACE group, n = 39) or followed by PA-TACE (PA-TACE group, n = 84) in our institution. Prognostic risk factors, disease-free survival (DFS), and overall survival (OS) were analyzed using the Cox proportional hazard regression model, the Kaplan-Meier method, and the log-rank test. Results: Liver cirrhosis was the only independent risk factor for SHCC patients. Overall, the PA-TACE group had no improved OS (P = 0.977) but worse DFS compared with the No TACE group (P = 0.045). Consistently, in subgroup analysis, SHCC patients with negative microvascular invasion (MVI), tumor size ≤ 5 cm and preoperative alpha-fetoprotein (AFP) < 400 ng/ml had similar OS (P = 0.466, P = 0.864, P = 0.488, respectively) but even worse DFS (P = 0.035, P = 0.040, P = 0.019, respectively) than those in the No TACE group. Besides, there was no significant difference in DFS and OS between the two groups of SHCC patients with liver cirrhosis (P = 0.342, P = 0.941, respectively). Conclusions: PA-TACE may not improve the long-term survival of SHCC patients, but may even potentially promote their postoperative tumor recurrence, especially for those with MVI-negative, tumor size ≤ 5 cm, and preoperative AFP < 400 ng/ml.

Erratum: TGF-ß induces growth suppression in multiple myeloma MM.1S cells via E2F1.

[This corrects the article DOI: 10.3892/ol.2017.6360.].

Circ-DENND4C up-regulates TCF4 expression to modulate hepatocellular carcinoma cell proliferation and apoptosis via activating Wnt/ß-catenin signal pathway.

Background: Hepatocellular carcinoma (HCC) is a common malignant tumor in China. Advanced treatment like transcatheter hepatic arterial chemoembolization (TACE) has prolonged the lives of many HCC patients. However, the prognosis of most HCC patients remains unsatisfactory. Recently, circular RNAs (circRNAs) have been gradually unveiled to exert considerable functions in cancer. Promising circRNAs in HCC remains to be further elucidated. Methods: Gene expression was assessed by qRT-PCR and western blot. The function of circ-DENND4C in HCC was estimated by both in vitro and in vivo experiments. The location of circ-DENND4C in HCC cells was determined by subcellular fractionation and FISH assays. The association among molecules were analyzed through RNA pull down, RIP and luciferase reporter assays. Results: circ-DENND4C (DENN domain containing 4C), an oncogene identified in breast cancer, was overexpressed in HCC cells. Also, circ-DENND4C exerted pro-tumor functions in HCC through activating Wnt/ß-catenin pathway. Importantly, circ-DENND4C could augment transcription factor 4 (TCF4) expression to activate Wnt/ß-catenin signaling via sequestering miR-195-5p. Moreover, following rescue assays disclosed that circ-DENND4C mediated malignant phenotypes in HCC cells via up-regulating TCF4 through sponging miR-195-5p. Conclusion: circ-DENND4C boosted TCF4 expression to modulate malignant behaviors of HCC cells via activating Wnt/ß-catenin pathway, which might offer a promising target for HCC treatment.

A new method of near-infrared fluorescence image-guided hepatectomy for patients with hepatolithiasis: a randomized controlled trial.

BACKGROUND: Hepatectomy is a definitive treatment for hepatolithiasis because it simultaneously removes intrahepatic duct (IHD) stones and biliary tract strictures together with the involved liver region en bloc. Unlike cystic or solid liver tumors, hepatolithiasis is usually associated with alterations of anatomical structures and perihepatic adhesions because of chronic recurrent inflammation. This complicates identification of the target hepatic region and location of biliary strictures. METHODS: To determine the efficacy of near-infrared fluorescence (NIRF) imaging using indocyanine green (ICG), we performed a comparative trial and developed a white-light and near-infrared dual-channel image-guided device (DPM-I) for both open and endoscopic surgery. Forty-four eligible patients were randomly assigned to Group A (NIRF imaging) or Group B (traditional hepatectomy). We injected ICG via peripheral veins for patients in Group A. RESULTS: The NIRF imaging method was associated with less blood loss (OR 1.004, 95% CI 0.999-1.010; P = 0.016), briefer hospitalization (OR 1.336, 95% CI 1.016-1.756; P = 0.001), lower rates of margins with dilated bile ducts (OR 1.278, 95% CI 1.030-1.585; P = 0.023), lower postoperative white blood cell counts (OR 1.262, 95% CI 0.931-1.712; P = 0.038), lower procalcitonin levels (OR 1.316, 95% CI 1.020-1.513; P = 0.002), and lower alanine aminotransferase levels (OR 1.013, 95% CI 1.003-1.023; P = 0.002) compared with traditional hepatectomy. CONCLUSIONS: These data demonstrate the efficacy of NIRF imaging with ICG using DPM-I for treating hepatolithiasis.

Long noncoding RNA HCG22 suppresses proliferation and metastasis of bladder cancer cells by regulation of PTBP1.

Multifarious biological functions of long noncoding RNAs (lncRNAs) have been reported in various cancers including bladder cancer (BCa). This study aims to determine the biological role of a certain lncRNA in BCa. Consistent with the data of The Cancer Genome Atlas database, it was validated that lncRNA HLA complex group 22 (HCG22) was weakly expressed in BCa samples and lowly expressed HCG22 was closely correlated with low overall survival of the BCa patient. To verify the role of HCG22 in BCa progression, functional experiments were carried out in two representative BCa cells (J82 and T24) and the negative effects of HCG22 expression on the cell proliferation, migration, and epithelial-mesenchymal transition were identified. Mechanistically, polypyrimidine tract-binding protein 1 (PTBP1), which was highly expressed in BCa tissues and cell lines, was negatively regulated by HCG22 and the PTBP1-mediated Warburg effect was also obstructed by HCG22. Furthermore, HCG22 modulated the expression of PTBP1 through destabilizing human antigen R (HuR). And functional rescue assays confirmed that HCG22 functioned in bladder cancer through downregulating PTBP1. In conclusion, the present study revealed that HCG22 inhibited BCa progression via the HuR/PTBP1 axis, opening new prospects for potent therapeutic regimens for BCa patients.

C14orf166 Is a Biomarker for Predicting Hepatocellular Carcinoma Recurrence.

Aim: Chromosome 14 open reading frame 166 (C14orf166) acts as a transcriptional repressor and is correlated with centrosome architecture manipulation. Nevertheless, the function of C14orf166 in hepatocellular carcinoma (HCC) progression remains unclear. We aimed to investigate the role C14orf166 plays in HCC and further compared the prognostic value of C14orf166 with that of clinicopathological features. Methods: C14orf166 expression was evaluated in a human liver cell line, HCC cell lines, HCC tissues and adjacent noncancerous liver tissues with qRT-PCR, western blot and immunohistochemistry. Patients were divided into two different groups according to C14orf166 level. The relationship between C14orf166 expression and clinicopathological features was assessed by Pearson chi-squared test and receiver operating characteristic curves. Cumulative disease-free survival (DFS) and overall survival (OS) curves were evaluated using the Kaplan-Meier method. Results: C14orf166 mRNA and protein expression is upregulated in HCC cell lines and tissues. The level of C14orf166 was correlated with serum alpha-fetoprotein level, lymph node metastasis, tumor size and recurrence, with high C14orf166 expression correlating with high HCC recurrence risk. The poor OS and DFS of HCC patients are partly due to the persistently high HCC recurrence risk. When combined with serum alpha-fetoprotein level, the predictive accuracy of C14orf166 for HCC recurrence was enhanced (AUC = 0.712, 95% CI 0.603-0.821; p = 0.001). Conclusions: This study demonstrated that C14orf166 is a high-risk biomarker and predictive factor for HCC recurrence, providing information for the selection of appropriate treatment strategies.

HIF-2α upregulation mediated by hypoxia promotes NAFLD-HCC progression by activating lipid synthesis via the PI3K-AKT-mTOR pathway.

Non-alcoholic fatty liver disease (NAFLD) is a relevant risk factor for developing hepatocellular carcinoma (HCC). Steatohepatitic HCC (SH-HCC), characterized by HCC with steatosis, is influenced by lipid metabolism disorders. A hypoxic microenvironment is common in HCC and affects lipid metabolism. However, whether hypoxia-induced HIF-2α upregulation exacerbates lipid accumulation to contribute to SH-HCC progression remains unclear. In this study, we demonstrated that HIF-2α was elevated in tissues from NAFLD-HCC patients and was associated with survival. Under hypoxic conditions, upregulated HIF-2α was accompanied by lipid accumulation and PI3K-AKT-mTOR pathway activation. HIF-2α knockdown (KD) in steatotic HCC ameliorated triglyceride accumulation and steatosis. HIF-2α-KD steatotic HCC showed minimal lipid synthesis in a hypoxic environment, which contributes to a reduction in malignant behaviours. However, treatment with MHY1485 restored these behaviours. STAM mice, a mouse model that develops NAFLD-HCC, exhibit more rapid tumour progression upon exposure to hypoxia. STAM mice treated with INK-128 presented abrogated mTOR expression and tumour progression under hypoxic conditions with lower triglycerides and steatosis. In conclusion, in a hypoxic microenvironment, HIF-2α upregulation promotes steatotic HCC progression by activating lipid synthesis via the PI3K-AKT-mTOR pathway. Therefore, HIF-2α can be a biomarker and target in developing specific therapeutic measures for NAFLD-HCC patients.

Nuclear GAPDH is vital for hypoxia-induced hepatic stellate cell apoptosis and is indicative of aggressive hepatocellular carcinoma behavior.

Background/aim: Hepatic stellate cells (HSCs) are critical determinants of liver tumor behavior such as vascular invasion, cell proliferation and migration. The apoptosis of HSCs can inhibit tumor growth and contribute to repressing hepatocellular carcinoma (HCC) progression. Our study aims to investigate the impact of nuclear glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on HSCs under hypoxic conditions and the association of nuclear GAPDH with HCC patient outcomes and tumor progression. Patients and methods: Following stable cell passage, 0.3% O2 was used to induce hypoxia. Cell proliferation and apoptosis were analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assays and flow cytometry, respectively. Proteins expression were detected by extracting nuclear and cytoplasmic proteins and performing Western blots. GAPDH nuclear translocation was blocked by the agent deprenyl. Immunohistochemical staining for GAPDH was investigated in 137 HCC tissue samples from our center. An analysis of the clinicopathological features, Kaplan-Meier analysis and Cox proportional hazards regression analysis were applied. Results: MTT assays and flow cytometry analyses showed that the nuclear accumulation of GAPDH led to the apoptotic death of HSCs, while blockade of this process with deprenyl significantly decreased apoptosis. Western blots revealed that deprenyl inhibited the nuclear translocation of GAPDH. An analysis of the immunohistochemical staining of HSCs in HCC tissue samples (137) revealed that nuclear GAPDH expression was significantly positively correlated with HIF-1α expression. Overall survival (OS) and time-to-recurrence (TTR) estimated by Kaplan-Meier analyses showed that patients with high HIF-1α or low nuclear GAPDH levels in HSCs had significantly poorer prognosis compared with patients with low HIF-1α or high nuclear GAPDH expression in HSCs. Moreover, patients with combined high HIF-1α/low nuclear GAPDH expression in HSCs had the worst prognosis. The Cox regression analysis revealed that the combination of nuclear GAPDH/HIF-1α expression in HSCs was an independent prognostic factor for OS and TTR in HCC patients. Conclusions: These findings provide a novel mechanism underlying the involvement of intranuclear GAPDH in hypoxia-induced HSCs apoptosis and a correlation between nuclear GAPDH levels and the clinical prognosis, which may prompt the development of a novel therapeutic strategy for HCC.

Lactobacillus rhamnosus GG Ameliorates Liver Injury and Hypoxic Hepatitis in Rat Model of CLP-Induced Sepsis.

BACKGROUND: Probiotic use to prevent gastrointestinal infections in critical care has shown great promise in recent clinical trials. Although well-documented benefits of probiotic use in intestinal disorders, the potential for probiotic treatment to ameliorate liver injury and hypoxic hepatitis following sepsis has not been well explored. METHODS: In order to evaluate, if Lactobacillus rhamnosus GG (LGG) treatment in septic rats will protect against liver injury, this study used 20-22-week-old Sprague-Dawley rats which were subjected to cecal ligation and puncture to establish sepsis model and examine mRNA and protein levels of IL-1ß, NLRP3, IL-6, TNF-a, VEGF, MCP1, NF-kB and HIF-1α in the liver via real-time PCR, Elisa and Western blot. RESULTS: This study showed that LGG treatment significantly ameliorated liver injury following experimental infection and sepsis. Liver mRNA and protein levels of IL-1ß, NLRP3, IL-6, TNF-a, VEGF, MCP1, NF-kB and HIF-1α were significantly reduced in rats receiving LGG. CONCLUSIONS: Thus, our study demonstrated that LGG treatment can reduce liver injury following experimental infection and sepsis and is associated with improved hypoxic hepatitis. Probiotic therapy may be a promising intervention to ameliorate clinical liver injury and hypoxic hepatitis following systemic infection and sepsis.

Exploiting Unlabeled Data in CNNs by Self-Supervised Learning to Rank.

For many applications the collection of labeled data is expensive laborious. Exploitation of unlabeled data during training is thus a long pursued objective of machine learning. Self-supervised learning addresses this by positing an auxiliary task (different, but related to the supervised task) for which data is abundantly available. In this paper, we show how ranking can be used as a proxy task for some regression problems. As another contribution, we propose an efficient backpropagation technique for Siamese networks which prevents the redundant computation introduced by the multi-branch network architecture. We apply our framework to two regression problems: Image Quality Assessment (IQA) and Crowd Counting. For both we show how to automatically generate ranked image sets from unlabeled data. Our results show that networks trained to regress to the ground truth targets for labeled data and to simultaneously learn to rank unlabeled data obtain significantly better, state-of-the-art results for both IQA and crowd counting. In addition, we show that measuring network uncertainty on the self-supervised proxy task is a good measure of informativeness of unlabeled data. This can be used to drive an algorithm for active learning and we show that this reduces labeling effort by up to 50 percent.

Clonorchiasis sinensis detected by laparoscopic exploration of biliary tracts in two patients with obstructive jaundice.

BACKGROUND: Hepatic clonorchiasis is one of the most prevalent food-borne parasitic diseases worldwide. Clonorchis sinensis, the pathogen, is the major parasitic trigger contributing to cholangitis, cholelithiasis, and even cholangiocarcinoma. Unfortunately, unspecific clinical manifestations of patients with hepatic clonorchiasis tend to mislead clinicians to neglect or misdiagnose them, following ignorance of appropriate therapy. Our case report may shed light on definite diagnosis of clonorchiasis with concomitant cholelithiasis, methodology for surgical drainage of the parasites, and postoperative anthelmintic therapy. CASE PRESENTATION: Two patients with habit of eating infected raw or undercooked freshwater fish were hospitalized due to right upper quadrant pain and jaundice. Magnetic resonance cholangiopancreatography (MRCP)/computed tomography (CT) detection indicated cholangiolithiasis and cholangiolithiasis with concurrent cholecystolithiasis, respectively. Fecal examinations were both negative for adult worms or eggs of parasites. However, adults of Clonrochis sinensis were detected within hepatobiliary tracts during laparoscopic cholecystectomy. Postoperative drainage and anthelmintic therapy contributed to complete recovery with good prognosis. CONCLUSIONS: Clonorchiasis provokes cholangiolithiasis and cholecystolithiasis. Standardized treatments for these gallstone patients with concomitant clonorchiasis include surgical removal of the calculus, postoperative T tubule drainage and anthelmintic therapy. Serological test or polymerase chain reaction (PCR)-based approaches might be helpful for diagnosis of clonorchiasis when no eggs are found by stool microscopy. Public health promotion on ceasing to eat raw freshwater fish is essential for prevention and control of clonorchiasis.

Kaempferol inhibits proliferation, migration, and invasion of liver cancer HepG2 cells by down-regulation of microRNA-21.

Liver cancer is one of the most common and lethal cancers in human digestive system, which kills more than half a million people every year worldwide. This study aimed to investigate the effects of kaempferol, a flavonoid compound isolated from vegetables and fruits, on hepatic cancer HepG2 cell proliferation, migration, invasion, and apoptosis, as well as microRNA-21 (miR-21) expression. Cell viability was detected using cell counting kit-8 (CCK-8) assay. Cell proliferation was measured using 5-bromo-2'-deoxyuridine (BrdU) incorporation assay. Cell apoptosis was assessed using Guava Nexin assay. Cell migration and invasion were determined using two-chamber migration (invasion) assay. Cell transfection was used to change the expression of miR-21. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to analyze the expressions of miR-21 and phosphatase and tensin homologue (PTEN). Expression of key proteins involved in proliferation, apoptosis, migration, invasion, and phosphatidylinositol 3-kinase/protein kinase 3/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway were evaluated using western blotting. Results showed that kaempferol significantly inhibited HepG2 cell proliferation, migration, and invasion, and induced cell apoptosis. Kaempferol remarkably reduce the expression of miR-21 in HepG2 cells. Overexpression of miR-21 obviously reversed the effects of kaempferol on HepG2 cell proliferation, migration, invasion, and apoptosis. Moreover, miR-21 negatively regulated the expression of PTEN in HepG2 cells. Kaempferol enhanced the expression of PTEN and inactivated PI3K/AKT/mTOR signaling pathway in HepG2 cells. In conclusion, kaempferol inhibited proliferation, migration, and invasion of HepG2 cells by down-regulating miR-21 and up-regulating PTEN, as well as inactivating PI3K/AKT/mTOR signaling pathway.

A novel 12-marker panel of cancer-associated fibroblasts involved in progression of hepatocellular carcinoma.

BACKGROUND/AIM: Cancer-associated fibroblasts (CAFs) are important factors in the progression of hepatocellular carcinoma (HCC). But the characterization of these cells remains incomplete. This study aims to identify a panel of markers for CAFs that are associated with HCC progression. MATERIALS AND METHODS: The sequencing data and clinicopathological characteristics of 366 patients were obtained from the Cancer Genome Atlas (TCGA) database (366 HCC tissues and there were 50/366 cases with corresponding normal liver tissues). In vitro validation of the markers was performed by quantitative real-time PCR using the hepatic stellate cell line LX2 induced by the HCC cell line Huh7. The activation of LX2 was confirmed by α-smooth muscle actin and fibroblast activation protein, using quantitative real-time PCR and immunofluorescence staining. In vivo detections of the 12 markers were done in 40 tissue samples (30 HCC and 10 normal). RESULTS: We successfully identified 12 CAF markers from TCGA data: FGF5, CXCL5, IGFL2, MMP1, ADAM32, ADAM18, IGFL1, FGF8, FGF17, FGF19, FGF4, and FGF23. The 12-marker panel was associated with the pathological and clinical progressions of HCC. All 12 markers were upregulated in vitro. In vivo expressions of these markers were paralleled with those in TCGA data. CONCLUSION: A 12-marker panel of CAFs in HCC is identified, which is associated with both pathological and clinical progressions of cancer.