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胸腺腫瘤(thymic tumor)是相當少見的腫瘤，90%發生於前縱膈腔。異位胸腺瘤(ectopic thymoma)發生率極低，好發部位多樣性，若無重症肌無力特徵性臨床表現，診斷困難。本文報告2位案例因甲狀腺超音波檢查發現疑似甲狀腺外結節，頸部電腦斷層顯示為低顯影、均質、邊界清楚之腫瘤。腫瘤上端與左葉甲狀腺下方邊界相連，腫瘤與甲狀腺組織呈現不同的顯影程度。2位個案皆接受自頸部路徑之腫瘤切除手術，最後病理報告皆證實為胸腺瘤AB型。根據Masaoka staging system與世界衛生組織(World Health Organization, WHO)分類，診斷為第二期胸腺瘤(thymoma)，WHO type AB。臨床上若表現為前頸部腫塊，無明顯臨床症狀，無重症肌無力症的表徵，甲狀腺超音波等影像學檢查除考慮甲狀腺本身常見疾病外，還要考慮到異位胸腺瘤可能。 Thymic tumors are rare neoplasms that commonly arise in the anterior mediastinum. Ectopic thymoma is even rarer. Aberrant or remnant ectopic thymic tissue can occur anywhere along the entire thymic descent pathway during embryogenesis in the neck or whole mediastinum, rendering diagnosis highly difficult to make. Herein we present 2 cases of ectopic thymoma presenting with an extra-thyroid nodule. Neck computed tomography (CT) revealed a hypodense and heterogeneous capsulated mass with smooth contour. The tumors were located in the anterior area of the neck and adjacent to the lower pole of the left side thyroid gland. These 2 cases both received anterior neck tumor excision. The mass was diagnosed as stage IIa and type AB thymoma based respectively on the Masaoka staging system and World Health Organization histological staging. Although it is well known that thymoma is often associated with myasthenia gravis, ectopic thymoma is seldom presented with myasthenic symptoms. Moreover, ectopic thymoma located in the neck region is apt to be misdiagnosed as a thyroid mass or other infiltrative lesion. Ectopic cervical thymoma presents a diagnostic pitfall especially on a fine needle aspiration cytology. The favored treatment protocol of ectopic thymoma is surgical resection, including an adequate surgical margin from the tumor. It is important to be aware that cervical masses may be thymomas rather than thyroid masses. &nbsp

Dengue virus (DENV) is a mosquito-borne pathogen that is becoming a serious global threat, owing to its rising incidence in inter-tropical regions that yield over 50 million annual infections. There are currently no approved antiviral agents for the management of dengue, and recent shortcomings in its immunization called for immediate action to develop effective drugs with prophylactic ability to better manage its infection. In an attempt to discover novel antiviral sources, we identified the medicinal herb Polygonum cuspidatum (PC) as a bioactive botanical material against DENV infectivity. Specifically, the methanolic extract from PC rhizomes (PCME) potently inhibited DENV infection without causing significant cytotoxicity. Further examination on the viral life cycle demonstrated that PCME particularly targeted the initial stages of DENV infection, while pre- and post-infection treatments had no effect. More importantly, the PCME could efficiently inactivate DENV free virus particles and block the viral attachment and entry/fusion events without apparently influencing viral replication, egress, and cell-to-cell spread. The antiviral effect of PCME was also recapitulated in infection analysis using DENV pseudoparticles displaying viral structural proteins that mediate DENV particle entry. Besides, PCME treatment also inhibited direct DENV entry into several cell types relevant to its infection and reduced viral infectivity of other members of the Flaviviridae family, including the hepatitis C virus (HCV) and Zika virus (ZIKV). Due to its potency against DENV entry, we suggest that the phytobioactive extract from PC is an excellent starting point as an antiviral source material for further development of therapeutic strategies in the prophylactic management of DENV infection.

We evaluated the analytical and clinical performance of a novel circulating tumor cell (CTC)-based blood test for determination of programmed death ligand 1 (PD-L1) protein expression status in real time in treatment-naïve non-small cell lung cancer (NSCLC) patients. CTCs were detected in 86% of patients with NSCLC (I-IV) at the time of diagnosis, with a 67% PD-L1 positivity rate (≥ 1 PDL + CTC). Among 33 NSCLC patients with PD-L1 results available via both tissue immunohistochemistry (IHC) and CTC assays, 78.9% were positive according to both methods. The CTC test identified an additional ten cases that were positive for PD-L1 expression but that tested negative via IHC analysis. Detection of higher PD-L1 expression on CTCs compared to that in the corresponding tissue was concordant with data obtained using other platforms in previously treated patients. The concordance in PD-L1 expression between tissue and CTCs was approximately 57%, which is higher than that reported by others. In summary, evaluation of PD-L1 protein expression status on CTCs isolated from NSCLC patients is feasible. PD-L1 expression status on CTCs can be determined serially during the disease course, thus overcoming the myriad challenges associated with tissue analysis.

Type 2 diabetes mellitus (T2DM) is a chronic disease characterized by hyperglycemia that can lead to long-term complications including heart diseases, stroke, retinopathy, and renal failure. Treatment strategies include stimulating glucose uptake and controlling blood glucose level. Bofutsushosan (BOF) and Daisaikoto (DAI) are two herb-based kampo medicines that have been demonstrated to improve metabolism-associated disorders including obesity, hyperlipidemia, and nonalcoholic fatty liver. Given their bioactivities against metabolic syndromes, we explored in this study the effect of BOF and DAI extracts on glucose absorption and used them as source to identify phytochemical stimulator of glucose absorption. Glucose uptake and mechanistic studies were evaluated in differentiated C2C12 skeletal muscle cells, and HPLC analysis was used to determine the molecular bioactive constituents. Our results indicated that the ethanolic extracts of BOF and DAI (BOFEE and DAIEE, respectively) enhanced the glucose uptake ratio in the differentiated C2C12 cells, and further analysis identified the flavone baicalin as a major constituent capable of efficiently stimulating glucose absorption. Mechanistic studies revealed that the effect from baicalin involved the activation of IRS-1 and GLUT-4, and implicated the AMPK, PI3K/Akt, and MAPK/ERK signaling cascades. Due to its potency, we suggest that baicalin merit further evaluation as a potential candidate anti-hyperglycemic agent for the treatment and management of T2DM. Keywords: Anti-hyperglycemia, Baicalin, Bofutsushosan, Daisaikoto, Glucose uptake

Enterovirus 71 (EV71) infection can cause airway symptoms, brainstem encephalitis, neurogenic shock, and neurogenic pulmonary edema with high morbidity and mortality. There is no proven therapeutic modality. Flos Farfarae is the dried flower bud of Tussilago farfara L. that has been used to manage airway illnesses for thousands of years. It has neuro-protective activity and has been used to manage neuro-inflammatory diseases. However, it is unknown whether Flos Farfarae has activity against EV71-induced neuropathy. The current study used both human foreskin fibroblast (CCFS-1/KMC) and human rhabdomyosarcoma (RD) cell lines to test the hypothesis that a hot water extract of Flos Farfarae could effectively inhibit EV71 infection. The authenticity of Flos Farfarae was confirmed by HPLC-UV fingerprint. Through plaque reduction assays and flow cytometry, Flos Farfarae was found to inhibit EV71 infection ([Formula: see text]). Inhibition of viral replication and protein expression were further confirmed by reverse transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR (qRT-PCR), and western blot, respectively. The estimated IC[Formula: see text]s were 106.3[Formula: see text][Formula: see text]g/mL in CCFS-1/KMC, and 15.0[Formula: see text][Formula: see text]g/mL in RD cells. Therefore, Flos Farfarae could be beneficial to inhibit EV71 infection by preventing viral replication and structural protein expression.

Phytochemical investigation of the ethanolic extract of Grangea maderaspatana led to isolation of gramaderins A-D, together with thirteen known compounds. All isolates were assayed for their anti-inflammatory activities. Consequently, 5,7-dihydroxy-3,6,3',4',5'-pentamethoxyflavone and 5,3'-dihydroxy-3,6,7,4',5'-pentamethoxyflavone showed significant bioactivities by inhibiting superoxide anion generation. 8-Acetoxy-pentadeca-1,9Z,14-trien-4,6-diyne-3-ol also demonstrated potent inhibition on elastase release. The gramaderins A/C (β-alkyl linked γ-lactone) and gramaderins B/D (α-alkyl linked γ-lactone) co-exist in this plant material, of which the latter derivatives are few in nature. Gramaderins C/D possess a special linear dilactone diterpene skeleton, which never been reported.

Ethnopharmacological relevance Bupleurum chinense is a traditional Chinese medicinal herb which has been used to treat various inflammatory and infectious diseases, while Bupleurum kaoi is an endemic plant in Taiwan. We determined whether B. chinense and B. kaoi and their biologically active saikosaponin compounds possess anti-melanoma activity. In addition, we developed a novel saikosaponin-d nanoparticle system to improve its solubility, and evaluated its antiproliferative effects and molecular mechanisms in melanoma cells. Materials and methods Ethanolic extracts from B. chinense and B. kaoi were prepared, and their saikosaponin contents were determined by high performance liquid chromatography analysis. Saikosaponin-d nanoparticles were synthesized, and their physicochemical properties were evaluated by particle size analyzer, transmission electron microscopy, differential scanning calorimetry, X-ray diffractometry, and Fourier transform infrared spectroscopy. Human A375.S2 melanoma cells were cultured, and cell viability determined by the MTT assay. Apoptosis was evaluated by determination of mitochondrial membrane potential, and signal transduction pathways investigated by Western blotting. Results Ethanolic extracts from B. kaoi showed more potent antiproliferative effect on human A375.S2 melanoma cells compared to B. chinense . The saikosaponin-a, -c and -d contents were higher in B. kaoi compared to B. chinense . Saikosaponin-d was the most potent compound in terms of anti-melanoma activity, and saikosaponin-d nanoparticles exhibited increased water solubility due to lowered particle size, amorphous transformation and intermolecular hydrogen bond formation with the excipient. Furthermore, saikosaponin-d nanoparticles showed enhanced antiproliferative activity against melanoma cells, and induced apoptosis through the mitochondrial pathway. The anti-melanoma activity was mediated by phosphorylation of JNK and p38, phosphorylation of p53, increased level of cytochrome c, and activation of caspase 9. Conclusions B. kaoi contains higher saikosaponin content and shows greater anti-melanoma activity than B. chinense . Saikosaponin-d nanoparticles have improved solubility, and may have potential use in the future as a form of treatment for melanoma.

Background Despite the advent of direct-acting antivirals (DAAs), HCV remains an important public health problem globally. There is at present no effective vaccine against the virus, and the DAAs in current use cannot prevent de novo infection, including in liver transplant setting wherein donor livers inevitably become re-infected. Developing inhibitors to HCV entry using nature-derived small molecules may help to expand/complement the current treatment options. Purpose In this study, we explored the effect of the plant alkaloid berberine (BBR) on HCV early viral entry. Methods Cell culture-derived HCV (HCVcc), viral pseudoparticles bearing HCV glycoproteins (HCVpp), and entry-related assays were employed to assess BBR's bioactivity. Molecular docking was used to predict BBR-HCV glycoproteins interaction, and the compound's antiviral activity was confirmed against HCVcc infection of primary human hepatocytes (PHHs). Results BBR specifically impeded HCVcc attachment and entry/fusion steps without inactivating the free virus particles or affecting the expression of host cell entry factors and post-entry viral replication. BBR also effectively inhibited infection by viral pseudoparticles expressing HCV E1/E2 glycoproteins and molecular docking analysis pointed at potential interaction with HCV E2. Finally, BBR could suppress HCVcc infection of PHHs. Conclusions We identified BBR as a potent HCV entry inhibitor, which merits further evaluation particularly for use in transplant setting against graft re-infection by HCV.

Two novel prenylated phenols, cyclocomunoindenol (1) and cyclocomunohexanol (2) were isolated from the cortex of roots of Artocarpus communis. Their structures were determined by spectroscopic methods. Compounds 2 and 6 revealed significant DPPH -scavenging activity with an IC50 values of 435.48 ± 0.93 and 53.55 ± 8.73 μM, respectively. Compounds 1, 2, and 6 displayed significant ABTS + scavenging activity with an IC50 values of 164.26 ± 2.44, 227.01 ± 3.64, and 44.09 ± 0.88 μM, respectively. Compound 6 exhibited an inhibitory effect on XO activity with an IC50 value of 10.91 ± 1.77 μM and the relative oxygen radical absorbance capacity (ORAC) values of 1–6, using ORAC-pyrogallol red (PGR) assay, were determined to be 0.28 ± 0.06, 0.31 ± 0.02, 0.55 ± 0.25, 0.84 ± 0.36, 0.35 ± 0.15, and 0.70 ± 0.09, respectively. Antioxidants 5 and 6 significantly attenuate UVA radiation-induced damage on human HaCaT keratinocytes and Hs68 fibroblasts. These finding showed that 1–6 may be used as antioxidants and 5 and 6 may protect skin against the adverse effects of UV radiation.

A novel benzoylphloroglucinol derivative, garcimultiflorone G (1), was isolated from the fruits of Garcinia multiflora. The structure of 1 was determined through extensive 1D- and 2D-NMR, and MS analyses. Garcimultiflorone G (1) showed inhibitory effects against superoxide anion (O·2(-) generation and elastase release by human neutrophils in response to formyl-L-methionyl-L-leucyl-L-phenylalanine/cytochalasin B (fMLP/CB), with IC50 values of 6.97 ± 1.56 and 11.70 ± 1.58 μM, respectively.

Yakammaoto is a prescription of traditional Chinese medicine (TCM) containing nine ingredients, including Ephedra sinica, Pinellia ternate, Zingiber officinale, Tussilago farfara, Aster tataricus, Ziziphus jujube, Belamcanda chinensis, Asarum sieboldii, and Schisandra chinensis. Yakammaoto has been used against flu-like symptoms for more than two thousand years in China and Japan. Coxsackievirus B4 (CVB4) causes not only flu-like symptoms but life-threatening diseases, such as pneumonia, acute kidney injury, and so forth with severe morbidity and mortality. There is no effective therapeutic modality against CVB4 infection. It is unknown whether yakammaoto is effective against CVB4 infection. We tested the hypothesis that yakammaoto can effectively inhibit CVB4-induced plaque formation in human airway and renal tubular cell lines by preventing viral attachment, internalization, and replication.The fingerprint of yakammaoto was assessed by HPLC. Effects of yakammaoto on CVB4 infection were tested by plaque reduction assay, reverse transcription polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA).Yakammaoto dose-dependently inhibited CVB4-induced plaque formation in HK-2, A549, and HEp-2 cells (p0.0001). Yakammaoto was both effective when supplemented prior to and after viral inoculation (p0.0001) by preventing viral attachment (p0.0001), internalization (p0.0001), and replication (p0.0001). Yakammaoto could decrease NGAL secretion before cytolysis to protect against viral injury.Yakammaoto had antiviral activity against CVB4-induced cellular injuries in airway mucosa and renal tubular epithelia by preventing viral attachment, internalization, and replication. The current study provides a basic support of its potential use against CVB4-induced airway and concomitant renal injuries.

In an effort to develop potent cyclooxygenase-1 (COX-1) inhibitors used as anticancer agent, a series of 2',5'-dimethoxychalcones was screened to evaluate their antiplatelet effect on human washed platelets suspension. Compound 2 exhibited potent inhibition of human washed platelet aggregation induced by collagen, significantly inhibited collagen- and arachidonic acid-induced thromboxane B2 release, and revealed inhibitory effect on COX-1 activity. Molecular docking studies showed that 1, 2, and 4 were bound in the active site of COX-1. These indicated that the antiplatelet effect of these compounds were mainly mediated through the suppression of COX-1 activity and reduced the thromboxane formation. To investigate the mechanistic action of COX-1 inhibitor enhanced the cytotoxic effect against human bladder cancer cells, NTUB1, we assessed the cytotoxic effect of 2 against NTUB1. Treatment of NTUB1 cells with various concentrations of 2 led to a concentration-dependent increase of cell death and decrease of reactive oxygen species levels. The flow-cytometric analysis showed that 2 induced a G1 phase cell cycle arrest but did not accompany an appreciable sub-G1 phase in NTUB1 cells. In addition, compound 2 increased p21 and p27 expressions and did not inhibit the expression of COX-1 in NTUB1 cells. Our results suggested that 2 enhanced cell growth inhibition or antiproliferative activity in NTUB1 cells through G1 arrest by COX-1 independent mechanism.

Ten new lantabetulic acid (1) derivatives 2-11 were synthesized and their cytotoxicities against human prostate cancer cells were evaluated. PC3 cells treated with 10 μM 8 exhibited the most potent G1 phase arrest. In addition, 10 μM 8 markedly decreased the levels of cyclin E and cdk2 and caused an increase in the p21 and p27 levels, while 20 μM 8 mainly led to cell death through the apoptotic pathway, which correlated with an increase in reactive oxygen species levels, decreased expression levels of Bcl-2 and caspase-8, the induction of mitochondrial changes, and decreased levels of cytochrome c in mitochondria. The dual action of 8 could provide a new approach for the development of chemotherapeutic drugs.

Bioassay-guided fractionation of the CHCl3 layer of Solanum violaceum areal parts methanolic extract led to the isolation of four new steroidal sapogenins, indiosides L-O (1-4), along with eight known steroids, one lignin, and a coumarin. Indioside L is a rare spirostanoid possessing a 1,4-dien-3-one moiety in ring A. Moreover, compounds 3 and 4 represent rare examples of spirostene with the 3β,7α-diol-5,6-ene moiety compared to the normal 3β,7β-diol-5,6-ene derivatives. The cytotoxic activity of the isolates (5-14) was evaluated against human hepatoma (HepG2 and Hep3B), human lung carcinoma (A549), and human breast carcinoma (MDA-MB-231 and MCF-7).

Without a vaccine, hepatitis C virus (HCV) remains a significant threat, putting 170–300 million carriers worldwide at risk of cirrhosis and hepatocellular carcinoma. Although the direct-acting antivirals targeting HCV replication have revolutionized the treatment of hepatitis C, several obstacles persist, including resistance development, potential side-effects and the prohibitive cost that limits their availability. Furthermore, treatment of HCV re-infection in liver transplantation remains a significant challenge. Developing novel antivirals that target viral entry could help expand the scope of HCV therapeutics and treatment strategies. Herein, we report (4R,6S)-2-dihydromenisdaurilide (DHMD), a natural butenolide, as an efficient inhibitor of HCV entry. Specifically, DHMD potently inhibited HCV infection at non-cytotoxic concentration. Examination on the viral life cycle demonstrated that DHMD selectively targeted the early steps of infection while leaving viral replication/translation and assembly/release unaffected. Furthermore, DHMD did not induce an antiviral interferon response. Mechanistic dissection of HCV entry revealed that DHMD could inactivate cell-free virus, abrogate viral attachment and inhibit viral entry/fusion, with the most pronounced effect observed against the viral adsorption phase as validated using ELISA and confocal microscopy. Due to its potency, DHMD may be of value for further development as an entry inhibitor against HCV, particularly for application in transplant setting.

Objective Silibinin is a flavonolignan that is well established for its robust antiviral activity against HCV infection and has undergone several clinical trials for the management of hepatitis C. Despite its potency, silibinin suffers from poor solubility and bioavailability, restricting its clinical use. To overcome this limitation, we developed highly bioavailable silibinin nanoparticles (SB-NPs) and evaluated their efficiency against HCV infection. Design SB-NPs were prepared using a nanoemulsification technique and were physicochemically characterised. Infectious HCV culture systems were used to evaluate the influence of SB-NP on the virus life cycle and examine their antioxidant activity against HCV-induced oxidative stress. The safety profiles of SB-NP, in vivo pharmacokinetic studies and antiviral activity against infection of primary human hepatocytes were also assessed. Results SB-NP consisted of nanoscale spherical particles ( 97% efficiency and increasing the compound9s solubility by >75%. Treatment with SB-NP efficiently restricted HCV cell-to-cell transmission, suggesting that they retained silibinin9s robust anti-HCV activity. In addition, SB-NP exerted an antioxidant effect via their free radical scavenging function. Oral administration of SB-NP in rodents produced no apparent in vivo toxicity, and pharmacokinetic studies revealed an enhanced serum level and superior biodistribution to the liver compared with non-modified silibinin. Finally, SB-NP efficiently reduced HCV infection of primary human hepatocytes. Conclusions Due to SB-NP9s enhanced bioavailability, effective anti-HCV activity and an overall hepatoprotective effect, we suggest that SB-NP may be a cost-effective anti-HCV agent that merits further evaluation for the treatment of hepatitis C.

Aim: Patients who undergo continuous ambulatory peritoneal dialysis (CAPD) frequently need second surgical procedures (SSP) for various reasons. After 18 years of practice and observation, we analysed the records of these patients and tried to determine what complications occurred most often and how to deal with them. Patients and methods: We retrospectively reviewed the medical records of patients receiving CAPD treatment in the past 18 years at our hospital. The impact of the most serious complications to the CAPD modality was presented by the technique survival rate. We also classified the SSP and analysed the indications for SSP. Results: All 206 SSP could be classified into 12 types of procedures. More than half of the procedures (51.9 per cent) were catheter removal, followed by debridement (14.1 per cent) and herniorrhaphy (10.2 per cent). There were 18 indications for SSP. The most common indication was peritonitis (21.4 per cent). The 3-year technique survival was only 23.2 per cent in 82 patients whose catheters were removed because of serious complications. Conclusions: During our long-term follow up of CAPD patients, more than one-third (36 per cent) needed SSP. Catheter removal forced by peritonitis still remains a major problem for patients and care providers. Technique survival was very low once a serious complication occurred.

The six new dihydro-β-agarofuranoid sesquiterpenes 1–6 and three known compounds were isolated from the whole plant of Celastrus paniculatus. The structures including relative configurations were elucidated by means of spectroscopic analyses. Compounds 1–6 were evaluated for cytotoxicity against a panel of three human-tumor cell lines.

Introduction The availability of targeted and immunotherapies has provided NSCLC patients with more effective treatment options. However, this has resulted in an increase in the number and modality of tests required for treatment selection. Given 30-50% of advanced lung cancer patients have insufficient or unavailable tissue for comprehensive genomic profiling, there is a need for a non-invasive assay that can accurately detect all guideline-recommended markers for NSCLC treatment selection. To meet this need, we have developed a blood test that detects six classes of alterations (SNV, Indels, Rearrangements, CNA, Microsatellite Instability and PD-L1 expression) for therapy selection. Methods & Results Three tubes of blood from a routine blood draw were sent to our CLIA-certified and/or CAP accredited lab for analysis. PD-L1 expression was evaluated in circulating tumor cells (CTCs) utilizing two different assays; (i) Immunofluorescence (IF) antibody staining, (ii) mRNA qPCR. CTCs were captured on the CMxTM CTC Platform coated with lipid bilayer and antibodies to EpCAM. PD-L1 expression results were highly correlated between IF and qPCR assays in ten solid tumor cell lines (lung, breast, prostate and colorectal cancer) spiked into whole blood to mimic the actual patient CTC capture process. In an ongoing study on clinical samples from NSCLC patients (N=20), we observed greater than 90% concordance between tissue (IHC by 22C3 PD-L1 clone) and blood (CTC IF and mRNA assays). A proprietary Single Molecule Sequencing (SMSEQTM) NGS assay was performed on plasma in order to detect 5 classes of genomic alterations (SNV, Indels, Rearrangements, CNA, MSI) from ctDNA. This assay was validated in accordance with the latest ACMG and AMP guidelines to accurately detect variants at low mutant allele fraction (.1% for SNVs and Indels, 1% for rearrangements and 5 copies for CNA) with high sensitivity and specificity. MSI status was determined by assessing nucleotide repeat sequences in five standard markers (BAT-25, BAT-26, MONO-27, NR-21, NR-24), and was detectable down to a MAF of 1%. In an ongoing study on clinical samples from NSCLC patients (N=20), we observed high concordance of MSI status between tissue (immunohistochemistry for dMMR/MSI status) and blood (ctDNA SMSEQ assay). Conclusion Tissue insufficiency and procurement challenges are the primary reasons why ~90% of patients diagnosed with advanced NSCLC are not comprehensively tested per NCCN-guidelines in the community setting where most cancer is treated, leading to suboptimal treatment selection. An accurate blood test that detects all 6 NCCN-recommended markers for immunotherapy and targeted therapy selection has the potential to significantly improve adherence to NCCN testing guidelines and enable optimal treatment selection. Citation Format: Huangpin B. Hsieh, Jen-chia Wu, Feng-Ming Lin, Julian Lucas, Alex Atkins, Pratyush Gupta, Hung-Jen Shao, Yen-Lin Chen, Wen-Jie Huang, Chia-Hsun Hsieh, Ruey Kuen Hsieh, Kuo-Wei Chen, Ming-Hong Yen, Mana Javey, Shih-En Chang, Twinkal Marfatia, Drew Watson, Mahul Amin, Ashish Nimgaonkar, Oscar Segurado, Rui Mei. CTC and ctDNA profiling to detect 6 NCCN-guideline recommended classes of alterations for immunotherapy and targeted therapy selection using sample from a single blood draw [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3635.

A new multiflorane triterpenoid and two new cucurbitane triterpenoids were isolated from the stems of Momordica charantia. The structures of the new compounds were elucidated by spectroscopic methods. These three new compounds, 1, 2 and 3 displayed ABTS radical cation scavenging activity with IC50 values of 268.5 ± 7.9, 352.1 ± 11.5 and 458.9 ± 13.0 μM, respectively and an inhibitory effect on xanthine oxidase (XO) activity with IC50 values of 142.3 ± 30.2, 36.8 ± 20.5 and 124.9 ± 8.3 μM, respectively. This finding indicated that 1–3 can be used as antioxidants.

Bioassay-guided fractionation of a methanol extract obtained from stems of Microtropis japonica led to the isolation of six new ursane-type triterpenoids (1-6) and a new 2,3-seco-oleanane-type triterpenoid (7), together with seven known compounds. The structures of the new compounds were elucidated using spectroscopic data analysis. Among the known compounds isolated, the main component, 8 (ursolic acid), was active for HL60 cells, and its effects on histone hyperacetylation and the inhibition of histone deacetylase (HDAC) activity were investigated.

Four new diterpenes, acasiane A ( 1), acasiane B ( 2), farnesirane A ( 3), and farnesirane B ( 4), along with three known diterpenes ( 5 - 7), two triterpenes ( 8 and 9), and eight flavonoids ( 10 - 17) were isolated from the roots of Acacia farnesiana. The structures and relative configurations of these compounds were determined by various spectroscopic and x-ray analyses. All isolated compounds were evaluated for their in vitro cytotoxic activities against six human cancer cell lines (Hep G2, Hep 3B, MDA-MB-231, MCF-7, A549, and Ca9 - 22) with the MTT method. Betulinic acid ( 8) displayed moderate cytotoxicity (1.70 - 5.74 microg/mL) towards five human cancer cell lines and the flavonoids had slight effects. In addition, 8, diosmetin ( 13), and 3',4',5-trihydroxy-7-methoxyflavone ( 15) slightly inhibited superoxide anion generation or elastase release by human neutrophils, indicating moderate anti-inflammatory activities.

Background and aim Huang-lian-jie-du-tang (HLJDT; Japanese name, oren-gedoku-to) is a traditional Chinese medicine prescription known to possess anti-inflammatory activity. Our study reports here for the first time the anticancer effect of HLJDT in two human liver cancer cell lines, Hep G2 and PLC/PRF/5. Methods Inhibition of cell proliferation by HLJDT was measured by sodium 3'-(1-(phenylamino-carbonyl)-3,4-tetrazolium)-bis(4-methoxy-6-nitro) benzene-sulfonic acid hydrate (XTT) assay. Clonogenic assay was used to elucidate the possible differences in long-term effects of HLJDT on human liver cancer cells. Cell cycle distribution was determined by flow cytometry. Apoptosis was detected using electrophoresis and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick endlabeling (TUNEL) assay. Protein expressions were determined by immunoblot assay. The activity of nuclear factor-kappa B (NF-kappaB) was determined by Trans-AM ELISA kit. In vivo tumor activity was assessed by xenograft study. Results HLJDT significantly increased the expression of inactivated phospho-Cdc2 and phospho-Cdc25C, and decreased the levels of cyclin A, cyclin B1, Cdc2, and Cdc25C, thereby contributing to cell-cycle arrest. HLJDT increased the expression of Bax and Bak, but decreased the level of Bcl-2 and Bcl-XL, and subsequently triggered the mitochondrial apoptotic pathway. In addition, HLJDT also inhibited cell-survival signaling by enhancing the amount of IkappaBalpha in the cytoplasm, reducing the level and activity of NF-kappaB in the nucleus, and subsequently attenuating the expression of Bcl-XL in Hep G2 and PLC/PRF/5 cells(.) The inhibitory effect mediated by HLJDT on cell growth was also demonstrated in a nude mouse model, in which the liver cancer cells induced tumor xenograft shrank considerably following treatment with HLJDT. Conclusions Taken together, these results suggest a potential anticancer effect of HLJDT against human liver cancer cells.

A new phloroglucinol, hyperielliptone HA (1/1a), a new spirophloroglucinol possessing an unprecedented skeleton, hyperielliptone HB (2/2a), and two new xanthonolignoids, hyperielliptones HC (3) and HD (4), were isolated from the heartwood of Hypericum geminiflorum. Compounds 1/1a and 2/2a were obtained as tautomeric pairs. The structures and relative configurations of these compounds were elucidated by spectroscopic methods. In biological testing, compound 2/ 2a revealed significant inhibition of oxidative DNA damage and an inhibitory effect on xanthine oxidase.

Without a vaccine, hepatitis C virus (HCV) remains a global medical and socio-economic burden, predisposing about 170 million carriers worldwide to end-stage liver diseases including cirrhosis and hepatocellular carcinoma. Although the recently developed direct-acting antivirals (DAAs) have revolutionized hepatitis C treatment, most of them are unsuitable for monotherapy due to risks of resistance, thus necessitating combination with interferon (IFN)-alpha, ribavirin, or additional DAAs. More importantly, the high cost associated with the DAAs restricts their accessibility to most parts of the world. Developing novel cost-effective anti-HCV therapeutics may help expand the scope of antivirals and treatment strategies against hepatitis C. Herein, we applied an activity-based and fraction-guided analysis of extracts from the medicinal plant Phyllanthus urinaria (P. urinaria), which yielded fraction 13 (F13) as possessing the most potent inhibitory activity against early viral entry of cell-culture HCV infection. Chemical analysis (silica gel chromatography followed by ESI LC-MS plus (1)H and (13)C NMR) of F13 identified loliolide (LOD), a monoterpenoid lactone, as a novel inhibitor of HCV entry. Specifically, LOD could efficiently inactivate HCV free virus particles, abrogate viral attachment, and impede viral entry/fusion, with minimal effect on viral replication/translation, particle production, and induction of type I IFN host antiviral immune response. ELISA-based binding analysis confirmed the monoterpenoid's ability in efficiently blocking HCV particle attachment to the host cell surface. Furthermore, LOD could inhibit infection by several genotypic strains of HCV. This is the first report characterizing P. urinaria and its bioactive compound LOD as potent HCV entry inhibitors, which merit further evaluation for development as candidate antiviral agents against hepatitis C.

For centuries, natural plant extracts have played an important role in traditional medicine for curing and preventing diseases. Studies have revealed that Artocarpus communis possess various bioactivities, such as anti-inflammation, anti-oxidant, and anticancer activities. A. communis offers economic value as a source of edible fruit, yields timber, and is widely used in folk medicines. However, little is known about its molecular mechanisms of anticancer activity. Here, we demonstrate the antiproliferative activity of A. communis methanol extract (AM) and its dichloromethane fraction (AD) in two human hepatocellular carcinoma (HCC) cell lines, HepG2 and PLC/PRF/5. Colony assay showed the long-term inhibitory effect of both extracts on cell growth. DNA laddering and immunoblotting analyses revealed that both extracts did not induce apoptosis in the hepatoma cell lines. AM and AD-treated cells demonstrated different cell cycle distribution compared to UV-treated cells, which presented apoptotic cell death with high sub-G1 ratio. Instead, acridine orange staining revealed that AM and AD triggered autophagosome accumulation. Immunoblotting showed a significant expression of autophagy-related proteins, which indicated the autophagic cell death (ACD) of hepatoma cell lines. This study therefore demonstrates that A. communis AM and its dichloromethane fraction can induce ACD in HCC cells and elucidates the potential of A. communis extracts for development as anti tumor therapeutic agents that utilize autophagy as mechanism in mediating cancer cell death.

Acute lymphoblastic leukemia (ALL), especially T-acute lymphoblastic leukemia (T-ALL), is a common childhood malignant neoplastic disorder. Chemotherapy agents, particularly those that can induce apoptosis, are the major intervening strategy in the treatment of ALL. In this study, we investigated in T-ALL cell line, CCRF-CEM, the in vitro cytotoxic effect and the mechanism of action of baicalin, a compound extracted from Scutellaria baicalensis Georgi and S. rivularis Benth (Labiateae). Results demonstrated that baicalin displayed a remarkable cytotoxic effect in CCRF-CEM, with an IC50value of 10.6 μg/ml. It triggered apoptotic effect by fragmentizing cellular DNA and arrested the cell cycle at G0/ G1phase. Baicalin (37.5 μg/ml)had not effected the expression of p53 and Fas protein. It was shown to decline the expression of Bcl-2 (22.0 pg/ml), which consequently caused the loss (52.7%)of transmembrane potential (ΔΨm) in the mitochondria after 72 hours of treatment. Baicalin (37.5 μg/ml) also elevated the amount of cytosolic cytochrome c (19.2 μg/ml), which finally triggered the activation of caspase-3 (50.1 pmol/min). In conclusion, baicalin was found to induce apoptosis in T-ALL cell lines through multiple pathways. This finding encourages further investigation of baicalin in its role as a potential candidate for chemotherapeutic agents in T-ALL.

In the present study, three materials extracted or isolated from the roots of B. kaoi, an endemic plant to Taiwan, were used to be examined the hepatoprotective effect against dimethylnitrosamine (DMN)-induced hepatic fibrosis in rats, they were water extract (BKW), polysaccharide-enriched fractions (BKP) and saponin-enriched fractions (BKS). After treated with DMN for 4 weeks, the levels of aminotrasferases (GOT, GPT) were significantly elevated in serum, and the levels of total protein (TP) and albumin were significantly decreased in serum and liver homogenates. Furthermore, the collagen contents were significantly elevated in liver homogenates and corresponded to the hepatofibrotic pathological examination. As the results showed, treated with groups of BKW, BKP, BKS markedly reduced GOT, GPT levels in rats serum. In addition, treated with groups of BKW, BKP, BKS markedly raised TP levels in rats serum and liver homogenates. Furthermore, treated with groups of BKW, BKP markedly raised albumin levels in rats serum and liver homogenates. Treated with groups of BKW, BKP, BKS markedly raised interferon-gamma (IFN-gamma) levels in rats serum, where only BKS and silymarin markedly raised interkeukin-10 (IL-10) levels in rats serum compared to that of DMN treated rats. None of test materials of B. kaoi except silymarin reduced the malondialdehyde (MDA) levels, but BKW, BKP markedly raised hepatic glutathione (GSH) levels to reveal the activity of anti-lipid peroxidation. Otherwise, treated with groups of BKW, BKP, BKS significantly reduced collagen contents in rats liver homogenates. In conclusion, B. kaoi demonstrated the anti-inflammatory and anti-fibrotic activities followed by anti-oxidant activity of enhanced GSH production, enhanced the liver cell regeneration and concerned with regulations of INF-gamma and IL-10. The ability of hepatoprotective and anti-fibrotic activities of B. kaoi are higher than B. chinense, a Bupleuri Radix imported from China to Taiwan.

Four new cerebrosides, gynuramides I˜IV (1-4), together with 37 known compounds were isolated from the rhizome of Gynura japonica. The structures of cerebrosides 1-4 were determined by chemical and spectroscopic examination to be: (2S,3S,4R,8E)-2-[(R)-2-hydroxypentacosanoylamino]-8-en-1,3,4-octadecanetriol, (2S,3S,4R,8E)-2-[(R)-2-hydroxytetracosanoylamino]-8-en-1,3,4-octadecanetriol, (2S,3S,4R,8E)-2-[(R)-2-hydroxytricosanoylamino]-8-en-1,3,4-octadecanetriol, and (2S,3S,4R,8E)-2-[(R)-2-hydroxydocosanoylamino]-8-en-1,3,4-octadecanetriol.

One new isoflavone, 5,7,4'-trihydroxy-2'-methoxyisoflavone (3) and seven, and four known compounds were isolated from the barks of Crotalaria pallida and the seeds of C. assamica, respectively. The known compounds, apigenin (1) and 2'-hydroxygenistein (2), isolated from C. pallida, showed significant concentration-dependent inhibitory effects on the release of beta-glucuronidase and lysozyme from rat neutrophils in response to formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB) with IC(50) values of 2.8+/-0.1 and 17.7+/-1.9, and 5.9+/-1.4 and 9.7+/-3.5 microM, respectively. The known compounds, daidzein (4) and 2'-hydroxydaidzein (6), isolated from C. pallida, inhibited of the release of lysozyme and beta-glucuronidase from rat neutrophils in response to fMLP/CB with IC(50) values of 26.3+/-5.5 and 13.7+/-2.6 microM, respectively. Compounds 1 and 4 also showed significant concentration-dependent inhibitory effects on superoxide anion generation in rat neutrophils stimulated with fMLP/CB with IC(50) values of 3.4+/-0.3 and 25.1+/-5.0 microM, respectively. Compounds 1 and 5, previously isolated from C. pallida, showed the inhibition of NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and LPS/interferon-gamma (IFN-gamma)-stimulated N9 microglial cells with IC(50) values of 10.7+/-0.1 and 13.9+/-1.1 microM, respectively. Flavonoids, suppressed chemical mediators in inflammatory cells, may have value in treatment and prevention of central and peripheral inflammatory diseases associated with excess production of chemical mediators.

One new pterocarpanoid, crotafuran E (1), and three known compounds were isolated from the bark of Crotalaria pallida. The structure of 1 was determined by spectral methods. Two pterocarpanoids, crotafurans A (2) and B (3), previously isolated from this plant, showed significant concentration-dependent inhibitory effects on the NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage-like cells, with IC(50) values of 23.0 +/- 1.0 and 19.0 +/- 0.2 microM, respectively. Compound 3 also inhibited the LPS/interferon-gamma (IFN-gamma)-stimulated NO production in N9 microglial cells with an IC(50) value of 9.4 +/- 0.9 microM. Moreover, compound 2 produced a concentration-dependent inhibition of the release of beta-glucuronidase and lysozyme from rat neutrophils in response to formyl-Met-Leu-Phe/cytochalasin B (fMLP/CB) with IC(50) values of 7.8 +/- 1.4 and 9.5 +/- 2.1 microM, respectively.

Extracts from natural plants have been used in traditional medicine for many centuries worldwide.Artocarpus communisis one such plant that has been used to treat liver cirrhosis, hypertension, and diabetes. To our knowledge, this study is the first to investigate the antihepatoma activity ofA. communistoward HepG2 and PLC/PRF/5 cells and the first to explore the relationship between antihepatoma activity and the active compound artocarpin content in different fractions ofA. communis.A. communismethanol extract and fractions induced dose-dependent reduction of tumor cell viability. DNA laddering analysis revealed thatA. communisextract and fractions did not induce apoptosis in HepG2 and PLC/PRF/5 cells. Instead, acridine orange staining revealed thatA. communistriggered autophagic cell death in a dose-dependent manner. The antihepatoma activity ofA. communisis attributable to artocarpin. The fractions with the highest artocarpin content were also the fractions with the highest antihepatoma activity in the following order: dichloromethane fraction > methanol extract > ethyl acetate fraction >n-butanol fraction >n-hexane fraction. Taken together,A. communisshowed antihepatoma activity through autophagic cell death. The effect was related to artocarpin content. Artocarpin could be considered an indicator of the anticancer potential ofA. communisextract.

Background & Aims A vaccine against hepatitis C virus (HCV) is unavailable and cost-effective antivirals that prevent HCV infection and re-infection, such as in the transplant setting, do not exist. In a search for novel and economical prophylactic agents, we examined the antiviral activity of saikosaponins (SSa, SSb2, SSc, and SSd) from Bupleurum kaoi root (BK) as entry inhibitors against HCV infection. Methods Infectious HCV culture systems were used to examine the effect of saikosaponins on the complete virus life cycle (entry, RNA replication/translation, and particle production). Antiviral activity against various HCV genotypes, clinical isolates, and infection of primary human hepatocytes were also evaluated. Results BK and the saikosaponins potently inhibited HCV infection at non-cytotoxic concentrations. These natural agents targeted early steps of the viral life cycle, while leaving replication/translation, egress, and spread relatively unaffected. In particular, we identified SSb2 as an efficient inhibitor of early HCV entry, including neutralization of virus particles, preventing viral attachment, and inhibiting viral entry/fusion. Binding analysis, using soluble viral glycoproteins, demonstrated that SSb2 acted on HCV E2. Moreover, SSb2 inhibited infection by several genotypic strains and prevented binding of serum-derived HCV onto hepatoma cells. Finally, treatment with the compound blocked HCV infection of primary human hepatocytes. Conclusions Due to its potency, SSb2 may be of value for development as an antagonist of HCV entry and could be explored as prophylactic treatment during the course of liver transplantation.

A novel compound containing an oxepane ring, hypertricone (1), was isolated from the leaves of Hypericum geminiflorum. Its structure, including the relative configuration, was elucidated from spectroscopic data and a computer-generated plot for the 3D structure.

A dichlorinated alkaloid with a novel skeleton, chingazumianine (1), was isolated from the herb Corydalis koidzumiana, and its structure elucidated by spectroscopic data and X-ray crystallographic analysis. In citrate-treated human platelet-rich plasma (PRP), known compounds, i.e., protopine (2), (±)-tetrahydropalmatine (3), and palmatine (4), isolated from this plant, showed significant inhibition of secondary aggregation induced by adrenaline in a concentration-dependent manner, suggesting that the antiplatelet effects of these compounds is mainly due to an inhibitory effect on thromboxane formation.

A novel phenolic compound, artocarpol A (1), was isolated from the root bark of Artocarpus rigida and its structure determined by spectroscopic methods and by comparison with its diacetate derivative. Compound 1 strongly inhibited superoxide formation in phorbol 12-myristate 13-acetate (PMA) stimulated rat neutrophils in a concentration-dependent manner with an IC50 value of 13.7±0.7 μM. Compound 1 also showed a significant inhibitory effect on tumor necrosis factor-α (TNF-α) formation in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells.