Exosome-derived miR-23a-5p inhibits HCC proliferation and angiogenesis by regulating PRDX2 expression: MiR-23a-5p/PRDX2 axis in HCC progression.

microRNAs (miRNAs) are closely related to the progression of hepatocellular carcinoma (HCC). Cancer-derived exosomes play an essential role in the establishment of the HCC microenvironment. However, the possible effects and underlying mechanisms of exosome (exo) microRNA-23a-5p (miR-23a-5p) in the progression of HCC remain unknown. In this study, we aimed to determine the role and specific molecular mechanism of exo miR-23a-5p in regulating HCC progression and to investigate whether exo miR-23a-5p levels can serve as an indicator of the prognosis of transarterial chemoembolization in patients with HCC. Our findings illustrated that miR-23a-5p was downregulated in exosomes separated from the serum of HCC patients and that miR-23a-5p carried by exosomes inhibited HCC cell proliferation and angiogenesis. Mechanistically, miR-23a-5p negatively targeted peroxiredoxin-2 (PRDX2). Functionally, PRDX2 overexpression relieved exosome-induced inhibition of HCC cell proliferation and angiogenesis by promoting vascular endothelial growth factor (VEGF) expression. In conclusion, Exo miR-23a-5p inhibited HCC proliferation and angiogenesis by regulating PRDX2 expression. Our results revealed the role and specific molecular mechanism of exo miR-23a-5p in regulating HCC progression.

The Predictive Role of Inflammatory Biomarkers for Treatment Response and Progression-Free Survival in Patients with Hepatocellular Carcinoma Receiving Hepatic Arterial Infusion Chemotherapy with FOLFOX Regimen: A Preliminary Study.

Objective: This retrospective study aimed to investigate whether pre-treatment inflammatory biomarkers, including the prognostic nutritional index (PNI), monocyte-lymphocyte ratio (MLR), systemic immune inflammation index (SII), and platelet-lymphocyte ratio (PLR), could predict treatment response and prognosis in patients with hepatocellular carcinoma (HCC) receiving hepatic arterial infusion chemotherapy (HAIC) with the oxaliplatin, leucovorin, and fluorouracil (FOLFOX) regimen. Methods: Based on the cut-off values identified using the receiver-operating characteristic (ROC) curve, 124 patients with HCC who received HAIC with the FOLFOX regimen were divided into low- and high-score MLR, PLR, PNI, and SII groups. Univariate and multivariate regression analyses were performed to identify independent predictors of treatment response and progression-free survival (PFS). Results: The cut-off values were 0.569 for MLR (area under the curve [AUC]: 0.621), 177.01 for PLR (AUC: 0.554), 713.05 for SII (AUC: 0.570), and 46.85 for PNI (AUC: 0.665). Multivariate Cox regression analysis revealed that the modified albumin-bilirubin (mALBI) grade (hazard ratio [HR]: 2.027; P=0.032), high MLR (HR: 7.250; P=0.002), and low PNI (HR: 0.296; P=0.003) were independent predictors of HAIC non-response, with an AUC value of 0.746 (95% CI: 0.658-0.833). A high MLR (HR: 1.714, 95% CI: 1.086-2.704, P=0.021) was also an independent predictor of PFS. Kaplan-Meier analysis showed that the patients with a high MLR had shorter PFS than those with a low MLR (median PFS: 6 vs 10 months, P=0.011). Conclusion: The pre-treatment MLR and PNI were predictors of non-response in patients with HCC receiving HAIC with the FOLFOX regimen. The MLR also was an independent predictor of PFS.

A Phase 1/2 Multicenter Randomized Trial of Local Ablation plus Toripalimab versus Toripalimab Alone for Previously Treated Unresectable Hepatocellular Carcinoma.

PURPOSE: To assess the safety and efficacy of local ablation plus PD-1 inhibitor toripalimab in previously treated unresectable hepatocellular carcinoma (HCC). PATIENTS AND METHODS: In the multicenter, two-stage, and randomized phase 1/2 trial, patients were randomly assigned to receive toripalimab alone (240 mg, every 3 weeks), subtotal local ablation followed by toripalimab starting on post-ablation day 3 (Schedule D3), or on post-ablation day 14 (Schedule D14). The first endpoint of stage 1 was to determine which combination schedule could continue and progression-free survival (PFS) as the primary endpoint for stage 1/2. RESULTS: A total of 146 patients were recruited. During stage 1, Schedule D3 achieved numerically higher objective response rate (ORR) than Schedule D14 for non-ablation lesions (37.5% vs. 31.3%), and was chosen for stage 2 evaluation. For the entire cohort of both stages, patients with Schedule D3 had a significantly higher ORR than with toripalimab alone (33.8% vs. 16.9%; P = 0.027). Moreover, patients with Schedule D3 had improved median PFS (7.1 vs. 3.8 months; P < 0.001) and median overall survival (18.4 vs. 13.2 months; P = 0.005), as compared with toripalimab alone. In addition, six (9%) patients with toripalimab, eight (12%) with Schedule D3, and 4 (25%) with Schedule D14 developed grade 3 or 4 adverse events, and one patient (2%) with Schedule D3 manifested grade 5 treatment-related pneumonitis. CONCLUSIONS: In patients with previously treated unresectable HCC, subtotal ablation plus toripalimab improved the clinical efficacy as compared with toripalimab alone, with an acceptable safety profile.

Differentiating Peripherally Located Pulmonary Noncalcified Hamartoma From Carcinoid Using CT Radiomics Approaches.

OBJECTIVE: This article aimed to differentiate noncalcified hamartoma from pulmonary carcinoid preoperatively using computed tomography (CT) radiomics approaches. MATERIALS AND METHODS: The unenhanced CT (UECT) and contrast-enhanced CT (CECT) data of noncalcified hamartoma (n = 73) and pulmonary carcinoid (n = 54; typical/atypical carcinoid = 13/41) were retrospectively analyzed. The patients were randomly divided into the training and validation sets. A total of 396 radiomics features were extracted from UECT and CECT, respectively. The features were selected by using the minimum redundancy maximum relevance and the least absolute shrinkage and selection operator to construct a radiomics model. Clinical factors and radiomics features were integrated to build a nomogram model. The performance of clinical factors, radiomics, and nomogram models on the differential diagnosis between noncalcified hamartoma and carcinoid were investigated. Diagnostic performance of radiologists was also explored. RESULT: In regard to distinguishing noncalcified hamartoma from carcinoid, the areas under the receiver operating characteristic curves of the clinical, radiomics, and nomogram models were 0.88, 0.94, and 0.96 in the training set UECT, and were 0.85, 0.92, and 0.96 in the training set CECT, respectively. The areas under the curve of the 3 models were 0.89, 0.96, and 0.96 in the validation set UECT, and were 0.79, 0.90, and 0.94 in the validation set CECT, respectively. The nomogram model exhibited good calibration and was clinically useful by decision curve analysis. Nomogram did not show significant improvement compared with radiomics, neither for UECT nor for CECT. Diagnostic performance of radiologists was lower than both radiomics and nomogram model. CONCLUSIONS: Radiomics approaches may be useful in distinguishing peripheral pulmonary noncalcified hamartoma from carcinoid. Radiomics features extracted from CECT provided no significant benefit when compared with UECT.

Apatinib Inhibits Angiogenesis in Intrahepatic Cholangiocarcinoma by Regulating the Vascular Endothelial Growth Factor Receptor-2/Signal Transducer and Activator of Transcription Factor 3/Hypoxia Inducible Factor 1 Subunit Alpha Signaling Axis.

INTRODUCTION: Intrahepatic cholangiocarcinoma (ICC), which is difficult to diagnose and is usually fatal due to its late clinical presentation and a lack of effective treatment, has risen over the past decades but without much improvement in prognosis. OBJECTIVE: The study aimed to investigate the role of apatinib that targets vascular endothelial growth factor receptor-2 (VEGFR2) in ICC. METHODS: MTT assays, cell scratch assays, and tube formation assays were used to assess the effect of apatinib on human ICC cell line (HuCCT-1) and RBE cells proliferation, migration, and angiogenic capacity, respectively. Expression of vascular endothelial growth factor (VEGF), VEGFR2, signal transducer and activator of transcription factor 3 (STAT3), pSTAT3, and hypoxia inducible factor 1 subunit alpha (HIF-1α) pathway proteins was assessed using Western blotting and mRNA expression analysis in HuCCT-1 was performed using RT-qPCR assays. The pcDNA 3.1(-)-VEGFR2 and pcDNA 3.1(-)-HIF-1α were transfected into HuCCT-1 and RBE cells using Lipofectamine 2,000 to obtain overexpressed HuCCT-1 and RBE cells. RESULTS: We found that apatinib-inhibited proliferation, migration, and angiogenesis of HuCCT-1 and RBE cells in vitro in a dose-dependent manner. We also proved that apatinib effectively inhibits angiogenesis in tumor cells by blocking the expression of VEGF and VEGFR2 in these cells. In addition, we demonstrated that apatinib regulates the expression of STAT3 phosphorylation by inhibiting VEGFR2. Finally, we showed that apatinib regulates ICC angiogenesis and HIF-1α/VEGF expression via STAT3. CONCLUSIONS: Based on the above findings, we conclude that apatinib inhibits HuCCT-1 and RBE cell proliferation, migration, and tumor angiogenesis by inhibiting the VEGFR2/STAT3/HIF-1α axis signaling pathway. Apatinib can be a promising drug for ICC-targeted molecular therapy.

[Ultrasound-guided microwave ablation with artificial pleural effusion for liver tumor adjacent to diaphragmatic dome].

OBJECTIVE: To explore the value of ultrasound-guided microwave ablation with artificial pleural effusion for liver tumor adjacent to diaphragmatic dome.
 Methods: A total of 34 patients with liver tumors located at diaphragmatic dome in Hunan Provincial Tumor Hospital were recruited from January 2014 to October 2015. The number of lesions ≤3 or lesion diameter ≤5 cm was in line with the microwave ablation indications. B ultrasound-guided microwave ablation for the liver tumors was undertaken after the artificial pleural effusion being established. 3-4 weeks later after the microwave ablation, all patients were imaged with enhance CT or MRI. The effect of ablation and the complications were evaluated.
 Results: There were 49 lesions in 34 patients, including 30 cases (88.2%) of complete ablation (CA), 3 cases (8.8%) of partial ablation (PA) and one case with new lesions after ablation (2.9%). Thirty-four patients had (1 580±230.7) mL of pleural effusion volume, while one case had bloody pleural effusion. One case had a diaphragmatic thermal injury, and one case had a biliary tumor infection. All of them showed remission after symptomatic treatment. 
 Conclusion: Combination of ultrasound-guided microwave ablation with artificial pleural effusion is a safe and effective therapy for liver tumor adjacent to diaphragmatic dome.

New approach for development of sensitive and environmentally friendly immunoassay for mycotoxin fumonisin B(1) based on using peptide-MBP fusion protein as substitute for coating antigen.

Here, on the basis of mimotope of small analytes, we demonstrated a new approach for development of sensitive and environmentally friendly immunoassay for toxic small analytes based on the peptide-MBP fusion protein. In this work, using mycotoxin fumonisin B1 (FB1) as a model hapten, phage displayed peptide (mimotope) that binds to the anti-FB1 antibody were selected by biopanning from a 12-mer peptide library. The DNA coding for the sequence of peptide was cloned into Escherichia coli ER2738 as a fusion protein with a maltose binding protein (MBP). The prepared peptide-MBP fusion protein are "clonable" homogeneous and FB1-free products and can be used as a coating antigen in the immunoassay. The half inhibition concentration of the quantitative immunoassay setup with fusion protein (F1-MBP and F15-MBP) was 2.15 ± 0.13 ng/mL and 1.26 ± 0.08 ng/mL, respectively. The fusion protein (F1-MBP) was also used to develop a qualitative Elispot assay with a cutoff level of 2.5 ng/mL, which was 10-fold more sensitive than that measured for chemically synthesized FB1-BSA conjugates based Elispot immunoassay. The peptide-MBP fusion protein not only can be prepared reproducibly as homogeneous and FB1-free products in a large-scale but also can contribute to the development of a highly sensitive immunoassay for analyzing FB1. Furthermore, the novel concept might provide potential applications to a general method for the immunoassay of various toxic small molecules.

Application of mimotope peptides of fumonisin b1 in Peptide ELISA.

Anti-fumonisin B(1) (FB(1)) McAb 1D11 was used as the target for biopanning from a phage random loop-constrained heptapeptide library. After three cycles of panning, seven phages with three mimotope peptides were selected to mimic the binding of FB(1) to 1D11. After the identification of phage ELISA, the phage clone that showed the best linear range of detection was chosen for further research. One peptide with the inserted peptide sequence of the phage was synthetized, named CT-452. An indirect competitive ELISA (peptide ELISA) for detecting FB(1) was established using the CT-452-bovine serum albumin conjugate as coating antigen. The linear range of the inhibition curve was 1.77-20.73 ng/mL. The half inhibitory concentration (IC50) was 6.06 ng/mL, and the limit of detection was 1.18 ng/mL. This method was compared with conventional indirect ELISA (commercial ELISA kit) and high-performance liquid chromatography (HPLC), and the results showed the reliability of the peptide ELISA for the determination of FB(1) in cereal samples. The relationship between the CT-452 and FB(1) standard concentrations in peptide ELISA was evaluated. The results indicated that synthetic peptide CT-452 can replace the FB(1) standard to establish an immunoassay free of FB(1).

Association between insulin-like growth factor-2 expression and prognosis after transcatheter arterial chemoembolization and octreotide in patients with hepatocellular carcinoma.

OBJECTIVE: To investigate the association between the change of IGF-2 level in serum after transcatheter arterial chemoembolization (TACE) and hepatocellular carcinoma (HCC) progression, especially in relation to metastasis. METHODS: IGF-2 in serum was measured by quantitative sandwich enzyme-linked immunosorbent assay before, 3 days and 4 weeks after TACE in 60 patients with HCC. The occurrence of HCC metastasis was also evaluated, 3 months after TACE. RESULTS: (1) The average serum level of IGF-2 in the 60 patients with HCC was 136.5 ± 87.3 pg/ml; (2) A tendency for increase was observed with heterogenous uptake of octreotide and portal vein thrombosis. Metastatic foci were found in 37/38 patients in the group with IGF-2 increasing (97.0%), in contrast to 3/22 (13.6%) patients with IGF-2 decrease. CONCLUSION: The increase of IGF-2 level in serum appears to be associated with the occurrence of metastatic HCC after TACE and chemotherapy.

[Transhepatic arterial chemoembolization with gemcitabine and carboplatin for the treatment of stage III hepatocellular carcinoma].

OBJECTIVE: To evaluate the efficiency and safety of transhepatic arterial chemoembolization (TACE) with gemcitabine and carboplatin for the treatment of stage III hepatocellular carcinoma (HCC). METHODS: Sixty-one HCC patients were treated by TACE. During TACE, At first, intra-arterial infusion of carboplatin 300 mg/m2, then gemcitabine 1000 mg/m2 with 5-30 ml of ultra-lipoidal iodide oil emulsion was used for arterial embolization. The toxicity and hepatic damage were observed according to WHO anticancer drug toxicity criteria and Child-Pugh classification criteria, respectively. The survival time was also observed during follow-up. RESULTS: The blood toxicity was bone marrow suppression presented as grade I leucopenia in 39.3%, grade II in 29.5%, grade III-IV in 18.0%. Grade II-III nausea and vomiting developed in 96.8% of the patients. Hepatic function damage became aggravated in 16 patients from A to B class, in 2 from A to C class, and in 6 from B to C class according to Child-Pugh classification criteria. The median survival time was 20 months with a range of 5 to 3 5 months. CONCLUSION: Transhepatic arterial chemoembolization using carboplatin and mixture of gemcitabine with ultra-lipoidal iodide oil emulsion is safe and effective in the management of stage III hepatocellular carcinoma. This regimen can also improve their quality of life.

Association between vascular endothelial growth factor and metastasis after transcatheter arterial chemoembolization in patients with hepatocellular carcinoma.

BACKGROUND: Hypoxia up-regulates vascular endothelial growth factor (VEGF) and stimulates the growth of hepatocellular carcinoma (HCC) cells. This study was designed to investigate the association between changes in plasma VEGF levels after transcatheter arterial chemoembolization (TACE) and HCC progression, especially in relation to metastasis. METHODS: Plasma VEGF levels were measured by quantitative sandwich enzyme-linked immunosorbent assay (ELISA R&D system). Plasma VEGF levels were measured before, 3 days and 4 weeks after TACE in 30 patients with HCC. The development of metastasis was evaluated at the end of the third month after TACE. RESULTS: The plasma VEGF levels of the 30 patients with HCC were 154.47+/-90.17 pg/ml. The total plasma VEGF levels after TACE increased compared with their basal levels (P<0.05), and the plasma VEGF levels had a tendency to increase in patients with heterogeneous uptake of iodizdoil and portal vein thrombosis. Follow-up for six months showed metastatic foci in 20 patients (74%) with increased plasma VEGF, but none of the patients with decreased plasma VEGF developed metastasis. CONCLUSION: Increased plasma VEGF expression is associated with the development of metastasis in HCC after TACE.

[Relation between vascular endothelial growth factor and reoccurence-metastasis after transcatheter arterial chemoembolization in hepatocellar carcinoma].

OBJECTIVE: To investigate the relation between changes in serum vascular endothelial growth factor (VEGF) level after transcatheter arterial chemoembolization (TACE) and hepatocellular carcinoma (HCC) progression, especially in relation to metastasis. METHODS: Serum VEGF expression level, measured by quatitative sandwich enzyme-linked immunosorbent assay (ELISA, R&D system), was measured before, 3 days and 4 weeks after TACE in 30 patients with HCC. The development of metastasis was evaluated at the end of the third month after TACE. RESULTS: 1. The serum VEGF level in 30 patients was 154.47 +/- 90.17 pg/ml, 2. Post-TACE total serum VEGF level increased as compared with their basal level in 30 patients (P < 0.05) and serum VEGF level had a tendency to increase in patients with heterogeneous uptake of iodized oil and portal vein thrombosis. During the follow-up of 1 - 2 years, metastatic foci were found in 74% (20) patients with SVEGF increase, while none of the patients showing SVEGF decrease developed metastasis. CONCLUSION: Serum VEGF expression increase is associated with the development of metastasis in hepatocellular carcinoma after TACE.