Evaluation of corn steep liquor as fermentation media for recombinant Lactococcus lactis producing antifreeze proteins.

BACKGROUND: Corn processing byproducts corn steep liquor (CSL), and thin stillage were evaluated as growth media for recombinant Lactococcus lactis modified to produce antifreeze proteins (AFPs) that could have important food and non-food applications. The AFP III sequence from ocean pout was cloned into a shuttle vector to make an expression vector that facilitated the production of recombinant AFP III in Lactococcus lactis. Light CSL from yellow dent corn and thin stillage from the industrial corn bioethanol process were optimized as fermentation media with a combination of the following additives and trace elements: disodium-ß-glycerophosphate (DG), tryptone (T), ascorbic acid (AA), iron (Fe), zinc (Zn), and magnesium (Mg). The growth of wild-type and recombinant Lactococcus lactis strains were compared over a 72 h period in 96-well plates and 250 mL shake flasks. RESULTS: The corn coproducts media consisting of 50% (v/v) light steep in water supplemented with DG-5 g L-1 , T-5 g L-1 , AA-0.5 g L-1 , and Zn-4 ppm resulted in best growth and was considered as the best-optimized media. The addition of additives and trace elements better supported the growth of both wild-type and recombinant Lactococcus lactis strains compared to control media without any additives. Respective fermentation supernatants were frozen to -20 °C, and the time to supercool and freeze was compared. A distinct supercooling effect was observed for the supernatants from recombinant strains thus, extending the time and temperature of supercooling and freezing. The maximum time of supercooling extended was 17.55 ± 4.45 min for thin stillage followed by M17 media (17.25 ± 4.45 min), Kent Corporation CSL (10.80 ± 2.12 min), and yellow dent CSL (6.9 ± 0.85 min) when fermented with recombinant Lactococcus lactis strains. CONCLUSION: The supplemented corn coproduct-based media enhanced the growth of both wild-type and recombinant Lactococcus lactis strains. These optimized media can replace or supplement more expensive media (e.g. M17), potentially reducing cost. The fermentation supernatants exhibited longer times to supercool, and freeze compared to control supernatants, indicating potential use as antifreeze compounds in frozen food and non-food applications. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Transcriptome profile and its partly verification of human hepatocellular carcinoma cells exposed to Yuzhizi () seed extract.

OBJECTIVE: To investigate the efficacy of Yuzhizi seed extract (FAQSE) on inhibiting the proliferation of hepatocellular carcinoma (HCC) cells in vitro and to explore the anti-HCC action mechanism of FAQSE. METHODS: Human HCC HepG2 and Huh7 cells were used to investigate the anti-HCC effect of FAQSE. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) method was used to measure cell viability. Affymetrix microarray was adopted to detect the expression of transcriptome. The differentially expressed genes (DEGs) of each cell line were identified. For co-DEGs of both cell lines, the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway were enriched using the Database for Annotation, Visualization and Integrated Discovery (DAVID), and the network analysis of protein-protein interaction (PPI) was mapped using the Retrieval of Interacting Genes/Proteins (STRING) and Cytoscape software. Some important genes in the PPI network of co-DEGs were selected to verify by quantitative real-time reverse transcription-polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay. RESULTS: FAQSE decreased the viability of HepG2 and Huh7 cells. There were 211 co-upregulated and 86 co-downregualted genes in both cell lines after FAQSE treatment. The enriched GO terms of co-upregulated DEGs were primarily involved cell-cell adhesion, viral process, transcription initiation from RNA polymerase II promoter, positive regulation of transcription from RNA polymerase II promoter and actin cytoskeleton organization. The GO terms of co-downregulated DEGs were mainly enriched in the processes of SRP-dependent cotranslational protein targeting to membrane, viral transcription, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, translational initiation and rRNA processing. Main KEGG pathways of co-upregulated DEGs were endocytosis, glutathione metabolism, protein processing in endoplasmic reticulum, synaptic vesicle cycle and lysosome. The major KEGG pathways of co-downregulated DEGs were ribosome, biosynthesis of amino acids, arginine and proline metabolism, systemic lupus erythematosus and complement and coagulation cascades. The top 10 co-DEGs with high hub nodes in STRING analysis were ribosomal protein S27a, transferrin, ribosomal protein S20, ribosomal protein L9, protein phosphatase 2 regulatory subunit B alpha, transthyretin, thioredoxin reductase 1, ribosomal protein L3, ribophorin I and ribosomal protein L24. Alpha-fetoprotein (AFP) was also co-downregulated and contained in the PPI network. The mRNA and protein expression of most verified genes was consistent with the results of co-DEGs analysis. And the AFP level was significantly reduced after FAQSE treatment. CONCLUSIONS: A series of genes and pathways of HepG2 and Huh7 cells were changed after FAQSE treatment, which might be the targets of FAQSE against HCC and worthy of further study. AFP might be important one of them.

Chemoprotective Effect of Decalactone on Hepatic Cancer via Diminishing the Inflammatory Response and Oxidative Stress.

Hepatocellular Carcinoma (HCC) is the 5th most common type of cancer in all types of cancers, globally. It is well known that the frequency of inflammatory reaction and oxidative stress increases during the HCC. The goal of this study was to see if decalactone could prevent rats against HCC caused by diethylnitrosamine (DEN). Single intraperitoneal administration of DEN (200 mg/kg) used as inducer and weekly intraperitoneal injection of phenobarbital (8 mg/kg) was used as promotor for induction the HCC in rats. Serum alpha fetoprotein (AFP) was used for the confirmation of HCC. Different doses of decalactone (5, 10 and 15 mg/kg) were orally administered to the rats. The body weight was determined at regular time. The hepatic, non-hepatic, antioxidant markers and inflammatory mediators were scrutinized. All groups of animals were scarified and macroscopically examination of the liver tissue was performed and the weight of organ (hepatic tissue) were estimated. Decalactone increased body weight while also suppressing hepatic nodules and tissue weight. Decalactone treatment reduced AFP, total bilirubin, and direct bilirubin levels while increasing albumin and total protein levels in a dose-dependent manner. Decalactone reduced lipid peroxidation (LPO) and increased catalase (CAT), glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels significantly (p < 0.001) (SOD). Decalactone lowered the levels of significantly (p < 0.001) inflammatory cytokines and inflammatory markers in the liver. Based on the findings, we may conclude that decalactone inhibited HCC in DEN-induced HCC animals via reducing oxidative stress and inflammatory mediators.

Treatment Patterns and Health Resource Utilization in Patients With Hepatocellular Carcinoma After Failure of Sorafenib in Real-World Setting in Taiwan.

OBJECTIVES: This study aimed to characterize current treatment patterns and healthcare resource utilization (HRU) observed among patients with hepatocellular carcinoma (HCC) after the failure of sorafenib in real-world setting in Taiwan. METHODS: A chart review was conducted in 130 patients; the inclusion criteria were patients with HCC who were aged 20 years or older and had received systemic therapy or best supportive care after failure of first-line systemic treatment with sorafenib between 2016 and 2018. Anonymized data on patient characteristics, treatment pathways, and survival were abstracted. RESULTS: The mean age of patients was 61.7 years (range 27-84); of these 130 patients, 103 (79%) were male, 81 (62%) had high alpha-fetoprotein (AFP) levels (≥400 ng/mL), and 96 (78.0%) were deceased at the time of data abstraction. After sorafenib therapy, 60 patients (46%) received systemic therapy, including nivolumab monotherapy (42%) and chemotherapy (25%). Oncologist visits at a semiannual per-patient rate of 3.7 (95% confidence interval [CI] 3.4-4.0) and hospitalizations at rate of 1.1 (95% CI 1.0-1.3) were the key contributors to HRU. Semiannual per-patient hospitalization rate was 1.3 (95% CI 1.1-1.5) in the high-AFP group. Median survival from discontinuation of sorafenib was 6.9 months (95% CI 5.9-9.0). CONCLUSIONS: This real-world evidence research on treatment patterns reflected substantial HRU consistent with the severity of HCC, particularly in the high-AFP group. Findings highlighted continuing high mortality in HCC, underlying a need for new treatments that can lengthen survival. Results can inform future evaluations of new HCC treatments that estimate the health economic impact of their adoption in Taiwan.

Simple Scoring System for Predicting TACE Unsuitable among Intermediate-Stage Hepatocellular Carcinoma Patients in the Multiple Systemic Treatment Era.

BACKGROUND/AIM: With the development of systemic treatment methods for unresectable hepatocellular carcinoma (uHCC), the concept of unsuitable for transcatheter arterial chemoembolization (TACE) has become important. This study aimed to establish a simple predictive scoring system for determining TACE unsuitable status. MATERIALS/METHODS: From 1998 to 2015, 196 patients with intermediate-stage uHCC with Child-Pugh A (score 5:6 = 108:88) and given TACE as the initial treatment were enrolled. At the baseline, tumor burden (Milan criteria-out, up-to-7 in/out, and up-to-11 in/out: 0-2 points) and modified albumin-bilirubin grade 1/2a or 2b (0-1 point) were added to determine the score for TACE unsuitable (CITRUS-MICAN score; low <2 and high ≥2). In addition, a previously reported tumor marker (TM) score, in which alpha-fetoprotein (AFP) was ≥100 ng/mL, fucosylated AFP ≥10%, and des-gamma-carboxy prothrombin ≥100 mAU/mL (each 1 point) (total 0, 1, or ≥2 points), was used for additionally evaluating tumor malignancy potential. Prognosis was retrospectively evaluated based on those scores. RESULTS: Median survival time (MST) was better for low compared to high CITRUS-MICAN score (42.0 vs. 26.4 months) (p = 0.002). A 2-step evaluation using the combination of CITRUS-MICAN and TM scores showed an MST of 43.2 months for low CITRUS-MICAN/TM score 0/1 (rank-A) and 39.6 months for low CITRUS-MICAN/TM score ≥2 (rank-B2), while it was 46.8 months for high CITRUS-MICAN/TM score 0 (rank-B1), 28.8 months for high CITRUS-MICAN/TM score 1 (rank-B2), and 22.8 months for high CITRUS-MICAN/TM score ≥2 (rank-C). For rank-A cases (n = 51), MST was 43.2 months, while it was 46.8 months for rank-B1 (n = 12), 31.2 months for rank-B2 (n = 82), and 22.8 months for rank-C (n = 51) (p = 0.001). CONCLUSION: The results showed that rank-C indicates absolute TACE unsuitable status. For rank-A patients, good prognosis with TACE can be expected, while TACE refractoriness status during the clinical course should be carefully evaluated so as to anticipate the appropriate timing for switching to systemic treatment in rank-B1 and -B2 patients.

Regorafenib for Taiwanese patients with unresectable hepatocellular carcinoma after sorafenib failure: Impact of alpha-fetoprotein levels.

BACKGROUND AND AIMS: Regorafenib has demonstrated its survival benefit for unresectable hepatocellular carcinoma (uHCC) patients in a phase III clinical trial. We aimed to assess the efficacy and tolerability of regorafenib and the predictors of treatment outcomes in Taiwanese patients. METHODS: We analyzed the survival, best overall response, predictors of treatment outcomes, and safety for uHCC patients who had tumor progression on sorafenib therapy and received regorafenib as salvage therapy between March 2018 and November 2020. RESULTS: Eighty-six patients with uHCC were enrolled (median age, 66.5 years; 76.7% male). The median regorafenib treatment duration was 4.0 months (95% confidence interval [CI], 3.6-4.6). The most frequently reported adverse events were hand-foot skin reaction (44.2%), diarrhea (36.0%), and fatigue (29.1%). No unpredictable toxicity was observed during treatment. The median overall survival (OS) with regorafenib was 12.4 months (95% CI, 7.8-17.0) and the median progression-free survival (PFS) was 4.2 months (95% CI, 3.7-4.7). Of 82 patients with regorafenib responses assessable, 4 patients (4.9%) achieved a partial response, and 33 (40.2%) had stable disease, leading to a disease control rate (DCR) of 45.1% (n = 37). Patients possessing baseline AFP < 400 ng/ml exhibited a markedly longer median OS, median PFS, and higher DCR compared with their counterparts (15.7 vs. 8.1 months, 4.6 vs. 3.7 months, 60.9% vs. 27.5%, respectively). Despite possessing high baseline AFP levels, patients with early AFP response (>10% reduction at 4 weeks or >20% reduction at 8 weeks after regorafenib administration) exhibited comparable treatment outcomes to those with baseline AFP < 400 ng/ml. CONCLUSIONS: The results of this real-world study verified the tolerability and efficacy of regorafenib treatment for uHCC patients who failed prior sorafenib therapy, especially for those with lower baseline AFP levels or with early AFP response.

Efficacy of Ramucirumab Versus Sorafenib as Subsequent Treatment for Hepatocellular Carcinoma.

BACKGROUND/AIM: The present study aimed to examine the therapeutic efficacy of ramucirumab compared with that of sorafenib as subsequent systemic therapy for patients with hepatocellular carcinoma (HCC) and serum α-fetoprotein (AFP) levels ≥400 ng/ml. PATIENTS AND METHODS: In our prospectively registered, real-world cohort, 13 and 11 patients treated with ramucirumab or sorafenib, respectively, were analyzed. Progression-free survival (PFS) was primarily compared between the ramucirumab and sorafenib groups. RESULTS: The PFS was significantly longer in the ramucirumab group than in the sorafenib group (median, 2.7 vs. 0.9 months, respectively; p=0.005). There were no significant differences in the objective response rates or the disease control rates between the ramucirumab and sorafenib groups (9.1% and 54.5% vs. 0.0% and 22.2%, respectively). CONCLUSION: Subsequent systemic therapy with ramucirumab showed a better ability to control tumor progression than sorafenib in HCC patients with serum AFP levels ≥400 ng/ml.

Lower Serum Sodium Levels Are Associated with the Therapeutic Effect of Sorafenib on Hepatocellular Carcinoma.

BACKGROUND AND AIM: Although the serum sodium level has been reported to be a prognostic and predictive marker for the therapeutic effects of lung cancer and renal cell carcinoma treated with molecular targeted therapy, the serum sodium level has not been investigated in hepatocellular carcinoma (HCC) patients treated with sorafenib. The aim of our analysis was to assess the prognostic role of serum sodium levels in these patients. METHODS: We retrospectively analyzed 341 HCC patients treated with sorafenib between 2009 and 2012 in our hospital and other related institutions. RESULTS: A total of 178 patients were enrolled in this study. The median age was 72 years (44-88), and 148 patients (83%) were male. The median overall survival (OS) was 12.9 months, and the median time to progression (TTP) was 3.1 months. Hyponatremia (hazard ratio (HR) 1.78, 95% confidence interval (CI) 1.26-2.52), a lower sodium level (HR 1.57, 95% CI 1.07-2.80), and a high level of α-fetoprotein (AFP) (≥ 200 ng/mL) (HR 1.78, 95% CI 1.26-2.52) were independent prognostic factors for TTP. We also categorized the patients into three groups according to serum sodium and AFP levels: Group A (n = 39) (serum sodium > 140 mEq/L, AFP < 200 ng/mL), Group C (n = 58) (serum sodium ≤ 140 mEq/L, AFP ≥ 200 ng/mL), and Group B (n = 81) (other patients). Significantly longer TTP and OS were observed in the following order: Groups A, C, and B. CONCLUSION: Serum sodium levels are associated with the effectiveness of sorafenib. The serum sodium level can predict the therapeutic effect of sorafenib in advanced HCC patients.

Thirty days oral Aframomum melegueta extract elicited analgesic effect but influenced cytochrome p4501BI, cardiac troponin T, testicular alfa-fetoprotein and other biomarkers in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Pain is the commonest symptom of a disease and the percentage of persons manifesting one form of pain is growing globally. Aframomum melegueta (AM) is commonly used by traditional doctors as medication for many ailments such as body pains and rheumatism because it possesses anti-inflammatory, anti-allergenic, antiviral, anti-ageing and anti-tumour phytochemical agents. AIM OF THE STUDY: Traditionally a botanical remedy in the management of pain was assessed. A common tropical plant Aframomum melegueta (AM) was evaluated for the amelioration of pain. For further pharmacologic understanding sensitive marker were used to assess the effect of the extract on the organ as a multifaceted approach to the evaluation of safety and analgesic efficacy. MATERIALS AND METHOD: Sensitive biomarkers such as troponin-T (CTnT), cardiac troponin-I (CTnI), interleukin-beta (IL-ß), interleukin-6 (IL-6), tumor necrosis factor-alfa (TNF-α) were evaluated using the enzyme-linked immunosorbent assay (ELISA) method and electrocardiographic parameters were also evaluated. The dynamics of concentrations of the various subfamilies of cytochrome were also assessed using ELISA in the evaluation of thirty-day oral AM, while histopathological changes of organs were also observed. RESULTS: Thirty-day oral AM doses 40 mg/kg and 80 mg/kg showed analgesic potential but influenced IL-6 level, IL-1ß, TNF-α and P-LCR. Electrocardiographic parameters showed the extract had arrhythmogenic effects the other cardiac parameters influenced was CTnT. The testicular alfa-fetoprotein and prostate specific antigen were also influenced. There were also some histopathological changes. CONCLUSIONS: The extract showed analgesic, anti-oxidant and anti-inflammatory potential with possible adverse effects consistent with testicular and prostate cancers, cardiovascular complication, hepatic congestion and cholestasis.

Serum Des-gamma-carboxy Prothrombin for Diagnosis of Adult Primary Cancer in Liver.

OBJECTIVE: To explore the diagnostic accuracy of serum des-gamma-carboxy prothrombin (DCP) in adult primary cancer in liver. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Department of Traditional Chinese Medicine, the First Affiliated Hospital, College of Medicine, Zhejing University, Hangzhou, China, from June 2016 to March 2018. METHODOLOGY: A retrospective study of 1,190 cases was undertaken and all the participants were divided into Group 1 (non-cancer patients) and Group 2 (primary cancer in liver patients). The records of DCP and alpha fetoprotein (AFP) levels were extracted and compared between Group 1 and Group 2. The sensitivity, specificity, and cutoff point of DCP in Group 2 and its different histological types were calculated. The receiver operating characteristic (ROC) curves were obtained, and the areas under the receiver operating characteristic curve (AUROC) were calculated and compared in different histological types. RESULTS: The DCP and AFP levels in Group 2 were markedly higher than Group 1 (p <0.001). For primary cancer in liver, the sensitivity, specificity, cutoff value, and AUROC of DCP were 64.2%, 91%, 56.5 mAU/mL, and 0.807 (95% CI: 0.783- 0.832), respectively. For hepatocellular carcinoma (HCC), the sensitivity, specificity, cutoff value, and AUROC of DCP were 69.3%, 92.2%,60.5mAU/mL, and 0.846 (95% CI: 0.817-0.875), respectively. For intrahepatic cholangiocarcinoma (ICC), the sensitivity, specificity, cutoff value, and AUROC of DCP were 13.6%, 93.2%, 76.5 mAU/mL, and 0.512 (95% CI: 0.439- 0.586), respectively. For combined hepatocellular-cholangiocarcinoma (cHCC-CC), the sensitivity, specificity, cutoff value, and AUROC of DCP were 66.7%, 81.5%, 40.5 mAU/mL, and 0.737 (95% CI: 0.585-0.888), respectively. CONCLUSION: DCP could be considered not only for surveillance and diagnosis of primary cancer in liver, but also for differential diagnosis of its different histological types.

Alpha-fetoprotein assessment for hepatocellular carcinoma after transarterial chemoembolization.

PURPOSE: To evaluate whether AFP classification criteria correlate with tumor response measured using the European Association for the Study of the Liver (EASL) and predicate survival in patients with hepatocellular carcinoma (HCC) after transarterial chemoembolization (TACE). METHODS: Data from 143 consecutive patients with unresectable HCC and elevated AFP (> 20 ng/mL), who underwent TACE as initial treatment between January 2011 and December 2015 were collected, retrospectively. AFP response was classified as follows: complete response, normalization of AFP; partial response, > 50% decrease from baseline; stable disease, - 50 to + 30% change from baseline; or progressive disease, > 30% increase from baseline. Response rates according to AFP and EASL criteria were compared, and associations between the AFP response and overall survival (OS) were evaluated. RESULTS: The k value for agreement between AFP criteria and EASL criteria was 0.52 (moderate), with response rates of 42.7% and 41.3%, respectively (P = 0.811). The OS of responders was significantly longer compared with non-responders for both AFP (21 vs. 6 months, P < 0.001) and EASL (23 vs. 6 months, P < 0.001). Multivariate analysis revealed that the AFP response (hazard ratio [HR], 0.430, 95% CI, 0.233-0.794; P = 0.007), EASL response (HR, 0.343; 95% CI, 0.176-0.666; P = 0.002), and macroscopic vascular invasion (HR, 2.104; 95% CI, 1.403-3.154; P < 0.001) were significantly associated with OS. CONCLUSIONS: The defined AFP classification criteria was moderate correlated with EASL criteria and predicted the outcome in patients with HCC who underwent TACE.

Transarterial Chemoembolization in Treatment-Naïve and Recurrent Hepatocellular Carcinoma: A Propensity-Matched Outcome Analysis.

OBJECTIVES: Transarterial chemoembolization (TACE) improves the survival of patients with hepatocellular carcinoma (HCC); however, TACE treatment outcomes of patients with treatment-naïve HCC (TN-HCC) and those with recurrent HCC after curative resection (R-HCC) have not yet been compared. METHODS: We recruited 448 patients with TN-HCC, and 275 patients with R-HCC treated with TACE as first-line anti-cancer treatment. RESULTS: At first TACE, patients with TN-HCC showed a significantly lower proportion of male gender (74.9% vs. 84.3%), higher proportion of liver cirrhosis (61.9% vs. 49.3%), higher aspartate aminotransferase (median 48 vs. 31 IU/L), alanine aminotransferase (median 38 vs. 26 IU/L), alpha-fetoprotein (AFP) (median 96.6 vs. 7.7 ng/mL), and total bilirubin (mean 1.0 vs. 0.8 mg/dL) levels, longer prothrombin time (median 1.05 vs. 1.01 international normalized ratio), higher tumor number (mean 2.1 vs. 1.7), larger tumor size (median 3.1 vs. 1.6 cm), and lower proportion of Barcelona Clinic Liver Cancer stage 0-A (55.6% vs. 71.9%) than patients with R-HCC (all P < 0.05). Multivariate analysis showed that TACE for TN-HCC (vs. R-HCC) was an independent predictor of mortality (hazard ratio, 1.328; P = 0.024) with AFP level and tumor number (all P < 0.05). However, treatment outcomes between TN-HCC and R-HCC became statistically similar after propensity score-matched (PSM) analysis using liver cirrhosis, tumor size, and multiple tumors (P < 0.05). CONCLUSIONS: Based on the similar TACE treatment outcomes observed with the PSM analysis, the current TACE treatment guideline for patients with TN-HCC might similarly be applied for patients with R-HCC.

Efficacy of anti-PD-1 antibody SHR-1210 as second-line treatment in hepatocellular carcinoma patient with sorafenib resistance: A case report.

RATIONALE: Hepatocellular carcinoma (HCC), one of the most common cancers worldwide, is an aggressive tumor with very poor prognosis. Regorafenib was the first agent to show a survival benefit over placebo in patients who showed progression while being treated with sorafenib, but it remains an unsatisfactory agent owing to its serious side effects. Therefore, more efficient and milder therapies are needed. PATIENT CONCERNS: Herein, we report a patient with advanced HCC with many lung metastases who showed progression during sorafenib treatment. DIAGNOSES: HCC with lung metastases (stage IVB). INTERVENTIONS: SHR-1210 alone was used as second-line treatment. OUTCOMES: Although the lung metastases did not decrease 3 months after the treatment, they decreased significantly at 6 months after the treatment and partially disappeared. The tumor response indicated partial response. Furthermore, all of the lung metastases continued to decrease at about 17 months after treatment. The alpha-fetoprotein levels showed a similar trend. After a follow up of 19 months, the patient remains in good health. LESSONS: SHR-1210 alone as a second-line treatment for a patient with HCC showed excellent antitumor effects. We think that SHR-1210 may exert its antitumor effects through a late-onset model, which persist for a long time. The side effects were mild and well tolerated.

Hepatocellular carcinoma macroscopic gross appearance on imaging: predictor of outcome after transarterial chemoembolization in a real-life multicenter French cohort.

BACKGROUND: Conventional transarterial chemoembolization (cTACE) with lipiodol is widely performed in patients with hepatocellular carcinoma (HCC) unsuitable for curative treatment. Additional tumor parameters such as HCC macroscopic appearance based on imaging might be helpful for transarterial chemoembolization prognostication and management. PATIENTS AND METHODS: A total of 405 patients with HCC who underwent cTACE between 2008 and 2016 from a real-life multicenter French cohort were retrospectively reviewed. Tumors were classified into two macroscopic types according to HCC gross appearance on imaging: nodular versus non-nodular. The study population was stratified into two groups: derivation and validation cohorts. Independent prognostic factors of survival based on multivariate cox regression models were determined and then assessed in the validation set. Thereafter, time to progression (TTP) and radiological response rate were investigated for each prognostic factors of survival. RESULTS: Median overall survival (OS) was 35 months for Barcelona Clinic Liver Cancer (BCLC) stage A, 22 months for BCLC stage B and 12 months for BCLC stage C patients (P < 0.0001). The corresponding TTP for these patients was 12 (7-17) months, 5 (3-6) months and 1.2 (1.2-3) months (P < 0.0001). Multivariate analysis revealed that tumors size and number, non-nodular type, alpha-fetoprotein, aspartate aminotransferase serum levels and impairment of performance status-1 were independent predictors of survival among the study groups. Non-nodular type was the most powerful factor that influences OS, TTP and radiological response rate for the recommended transarterial chemoembolization candidates. TTP was consistent with OS within each stage. CONCLUSION: HCC macroscopic appearance on imaging is a determinant predictor of outcome after cTACE in a real-life multicenter cohort.

Alpha-fetoprotein in animal reproduction.

Alpha-fetoprotein (AFP) is a serum glycoprotein with structural and physico-chemical properties similar to albumin. However, the exact physiological functions of AFP remain unknown; those known to date include markers to pathological conditions including neoplastic and non-neoplastic diseases, antioxidant effects, growth regulator in different cells and in cancer, immune response modulator, and carrier for fatty acids and oestrogens. This review aimed to present an overview of the different functions of AFP, particularly its role in the sexual differentiation of the hypothalamus, because its ability to bind oestrogens prevents their passage to the brain, where they inhibit the surge centre development. AFP and anti-Mullerian hormone are known to be involved in the development of freemartins, or genetically female foetuses masculinised in the presence of a male co-twin.

Prognostic predictors for patients with hepatocellular carcinoma receiving adjuvant transcatheter arterial chemoembolization.

OBJECTIVES: We aimed to confirm the clinical effectiveness of postoperative adjuvant transcatheter arterial chemoembolization (PA-TACE) in patients with hepatocellular carcinoma after liver resection, and further identify the patients who could benefit most from PA-TACE. PATIENTS AND METHODS: Propensity score matching at a ratio of 1 : 2 was used between hepatectomy patients with and without receiving PA-TACE. Kaplan-Meier analysis was performed to compare overall survival and recurrence-free survival between two groups. Univariate COX regression and stratified analyses were performed to screen and identify survival predictors for PA-TACE patients. The identified predictive markers were validated in an external cohort. RESULTS: The propensity analysis matched 116 patients in PA-TACE group to 232 in the control group. Visible protective effect of PA-TACE was shown by survival curves in matched series (log-rank P=0.009 and 0.008), with hazard ratio of being 0.599 (95% confidence interval: 0.420-0.855) and 0.623 (95% confidence interval: 0.449-0.866), respectively, for overall survival and recurrence-free survival. The identified prognostic predictors for PA-TACE included TNM stage, tumor size and number, hepatitis B infection, spleen diameter, preoperative serum α-fetoprotein, alkaline phosphatase, γ-glutamyl transpeptidase and monocyte, and three risk signatures (aspartate aminotransferase-to-alanine aminotransferase ratio, neutrophil-to-lymphocyte ratio, and systemic immune-inflammation index). CONCLUSION: The treatment effectiveness of adjuvant transcatheter arterial chemoembolization for patients with hepatocellular carcinoma after surgery was validated in this study, and the best candidates for PA-TACE were identified as well, including patients with late-stage tumor, portal hypertension, and high preoperative serum levels of α-fetoprotein, alkaline phosphatase, γ-glutamyl transpeptidase, and monocytes.

A feasibility study of the toxic responses of human induced pluripotent stem cell-derived hepatocytes to phytochemicals.

Herbal medicines have been increasingly used in the last three decades. Despite their popularity, safety issues with herbal products need to be addressed. We performed a feasibility study of the toxic responses of human induced pluripotent stem cell-derived hepatocytes (iHep cells) to phytochemicals in comparison with hepatoblasoma-derived HepG2 cells and long-term human hepatocytes (LTHHs). The iHep cells expressed typical hepatocyte markers cytochrome P450 3A4 (CYP3A4), hepatocyte nuclear factor 4α, and albumin despite the expression of immature markers α-fetoprotein and cytokeratin 19. We studied the responses of iHep cells to phytochemicals saikosaponin D, triptolide, deoxycalyciphylline B, and monocrotaline with different mode of toxicity employing MTS and lactate dehydrogenase (LDH) assays. Saikosaponin D and triptolide caused dose-dependent cytotoxicity in the iHep cells, which were more sensitive than LTHHs and HepG2 cells. Saikosaponin D-induced cytotoxicity tightly correlated with increased LDH leakage in the iHep cells. Although deoxycalyciphylline B did not exhibit toxic effect on the iHep and HepG2 cells when compared with LTHHs, it decreased CYP3A7 expression in the iHep cells and increased CYP1A2 expression in HepG2 cells. We hereby show the feasibility of using iHep cells to detect toxic effects of phytochemicals.

Rheum palmatum root extract inhibits hepatocellular carcinoma in rats treated with diethylnitrosamine.

OBJECTIVES: The aim of this study was to investigate the potential anticancer properties of a methanol extract of Rheum palmatum roots against diethylnitrosamine (DENA)-induced hepatocellular carcinoma (HCC) in rats and to characterize its phytoconstituents. METHODS: HPLC-PDA-MS/MS was used to profile the secondary metabolites in R. palmatum root extract. HCC was induced using diethylnitrosamine (DENA). The activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT), alpha-fetoprotein (AFP), total proteins, serum albumin and serum globulin was determined. DNA fragmentation and histopathological examination and GST-P immunostaining were also studied. KEY FINDINGS: LC-MS/MS analysis identified 16 compounds belonging to anthraquinones, flavonoids and tannins. The root extract significantly reduced the elevated liver enzymes ALT and AST and increased total proteins, albumin and globulin in HCC-rats. Also, the tumour markers AFP and GGT levels were significantly reduced in HCC-rats treated with the extract. In addition, the extract significantly reduced elevated DNA fragmentation and decreased the numbers and areas of GST-P positive putative foci in HCC-rats. CONCLUSIONS: Rheum palmatum is a potential candidate to be explored for the treatment of hepatocellular carcinoma.

The outcome of sorafenib monotherapy on hepatocellular carcinoma with portal vein tumor thrombosis.

Sorafenib is not recommended for advanced hepatocellular carcinoma (HCC) patients with Vp4 (portal invasion at the main trunk) by the Japan Society of Hepatology (JSH) due to a risk of hepatic failure. This study aimed to elucidate the safety and efficacy of sorafenib monotherapy on HCC with macro-vascular invasion (MVI). A total of 415 consecutive advanced HCC patients received sorafenib in our hospital. Patients with only MVI and sorafenib monotherapy were retrospectively enrolled. We enrolled 113 (27.2%) patients, including 56 (49.5%) Vp3 (portal invasion at the first branch) and 57 (50.5%) Vp4. Their median intervals of follow-up and sorafenib-use were 7.8 months and 2.7 months respectively. Using sorafenib, more Vp4 had hepatic decompensation (HD) (37% VS 18.2%, p = 0.028) than Vp3 patients. The multivariate analysis showed Vp4 (Odds ratio: 2.91; 95% CI: 1.02-8.3, p = 0.041) and baseline alpha-fetoprotein (AFP) ≥ 200 ng/ml were associated with HD. Dividing our patients into four subgroups as Vp3 + AFP < 200 ng/ml, Vp3 + AFP ≥ 200 ng/ml, Vp4 + AFP < 200 ng/ml and Vp4 + AFP ≥ 200 ng/ml, the proportions of HD were 16.7%, 19.4%, 16.7% and 55.2% respectively (p = 0.002). The overall survival rates were distributed with a significant decreasing trend as 10.2 ± 4.4 months, 6.5 ± 1.0 months, 6.0 ± 1.3 months and 2.5 ± 0.5 months (p = 0.001). We found only Vp4 plus AFP ≥ 200 ng/ml could induce more HD and a poorer prognosis than Vp3 patients. Hence, in Vp4 patients with higher AFP, sorafenib should not be the first-line treatment due to its limited survival benefit.

Elevation of tumour markers TGF-ß, M2-PK, OV-6 and AFP in hepatocellular carcinoma (HCC)-induced rats and their suppression by microalgae Chlorella vulgaris.

BACKGROUND: Chlorella vulgaris (ChV), a unicellular green algae has been reported to have anticancer and antioxidant effects. The aim of this study was to determine the chemopreventive effect of ChV on liver cancer induced rats by determining the level and expression of several liver tumour markers. METHODS: Male Wistar rats (200-250 g) were divided into 4 groups according to the diet given: control group (normal diet), ChV group with three different doses (50, 150 and 300 mg/kg body weight), liver cancer- induced group (choline deficient diet + 0.1% ethionine in drinking water or CDE group), and the treatment group (CDE group treated with three different doses of ChV). Rats were killed at 0, 4, 8 and 12 weeks of experiment and blood and tissue samples were taken from all groups for the determination of tumour markers expression alpha-fetoprotein (AFP), transforming growth factor-ß (TGF-ß), M2-pyruvate kinase (M2-PK) and specific antigen for oval cells (OV-6). RESULTS: Serum level of TGF-ß increased significantly (p < 0.05) in CDE rats. However, ChV at all doses managed to decrease (p < 0.05) its levels to control values. Expressions of liver tumour markers AFP, TGF-ß, M2-PK and OV-6 were significantly higher (p < 0.05) in tissues of CDE rats when compared to control showing an increased number of cancer cells during hepatocarcinogenesis. ChV at all doses reduced their expressions significantly (p < 0.05). CONCLUSIONS: Chlorella vulgaris has chemopreventive effect by downregulating the expression of tumour markers M2-PK, OV-6, AFP and TGF-ß, in HCC-induced rats.