Antioxidant and Anti-diabetic Properties of Aqueous Methanol Extraction of Nephelium mutabile (Pulasan) Leaves

2020 ◽  
Vol 2 (1) ◽  
pp. 27
Keyword(s):  
Column Chromatography ◽  
Inhibitory Activity ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Alpha Amylase ◽  
Total Phenolic ◽  
Dpph Assay ◽  
Aqueous Methanol ◽  
Ic50 Value ◽  
Alpha Glucosidase

Nephelium mutabile is a tropical plant of Sapindaceae family which originated from the Malay Peninsula and used in traditional medicine. This study focused on investigating the antioxidant and anti-diabetic properties of Nephelium mutabile leaves. The aqueous methanol (80%) crude extract of Nephelium mutabile leaves were fractionated via gravity column chromatography and thin-layer chromatography. A total of 17 fractions were obtained from column chromatography and subsequently pooled together to yield eight fractions. The pooled fractions were examined for their antioxidant properties via DPPH assay, Folin-Ciocalteu assay, and aluminum chloride colorimetric assay and tested for anti-diabetic properties using alpha-amylase and alpha-glucosidase inhibitory assay. DPPH assay showed that fraction 8 has the highest free radical scavenging activity (EC50 =88.0 μg/mL) followed by fraction 7 (EC50 =225.0 μg/mL). Fraction 8 showed the highest total phenolic and flavonoid content (221.1 μg GAE/mg, 222.4 μg QE/mg) respectively followed by fraction 7 (166.5 μg GAE/mg, 213.9 μg QE/mg) respectively. The most potent alpha-amylase inhibitory activity is observed in fraction 8 with an IC50 value of (41.0 μg/mL) followed by fraction 7 (90.6 μg/mL). In the alpha-glucosidase inhibitory activity, fraction 8 with IC50 value 160.0 μg/mL followed by fraction 7 with IC50 value 50.6 μg/mL showed the best inhibition activity. In conclusion, Nephelium mutabile leaves showed potential antioxidant, and anti-diabetic properties and fractions 8 and 7 should be the focus of future studies.

scholarly journals In Vitro Antioxidant and Antiproliferative Efficacy of Abrus Precatorius Seed Extracts on Hep2c Cells and its Inhibitory Efficacy with Receptors of Cervical Carcinoma

2022 ◽  
Author(s):  
Amritpal Kaur ◽  
Yash Sharma ◽  
Kamran Waidha ◽  
Madhumita P Ghosh ◽  
Anoop Kumar ◽  
...  
Keyword(s):  
Tannic Acid ◽  
Herbal Remedy ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Seed Extract ◽  
Total Phenolic ◽  
Cervix Carcinoma ◽  
Abrus Precatorius ◽  
Ic50 Value ◽  
Seed Extracts

Abstract Abrus precatorius is a widely distributed tropical medicinal plant with several therapeutic properties, however its seed extract has not been studied against cervical cancer (CaCx) till date. Herein, we have assessed the antioxidant and antiproliferative properties of A. precatorius seed extracts (ethyl acetate and 70% ethanol) prepared from different extraction methods (Soxhlet and maceration) against human cervix carcinoma cells (Hep2C). We observed a significantly higher total flavonoid content of APE (sox) i.e.,112.7±0.127 mg Quercetin Equivalent/g of extract than others; total phenolic content of APA (mac) seed extract was higher i.e.,108.53±0.089 mg Quercetin Equivalent/g of extract and total tannin content of APA (sox) was higher i.e., 98.98± 0.011 mg tannic acid equivalents/g of extract. In addition, tannic acid, rutin and piperine were identified in extracts by HPLC. Furthermore, APA (sox) exhibited the highest radical scavenging activity with an IC50 value of 14.49±0.93µg/ml. APE (mac) showed the most significant antiproliferative activity with IC50 value of 85.90±0.93µg/mL against Hep2c cells. SOD and GST activity was observed as highest in the APA(Mac) extract whereas the catalase activity and GSH content was maximum in the APE(Mac) extract. Further, the MDA content was observed to be the least in APE(Mac) extracts. Docking results suggested maximum binding energy between tannic acid and Her2 receptor compared to doxorubicin (standard). This study provides evidence that A. precatorius seed extracts possess promising bioactive compounds with probable anticancer and antioxidant properties against CaCx which might be utilized as a possible herbal remedy aimed at restricting tumor growth.

scholarly journals The Combination of Mild Salinity Conditions and Exogenously Applied Phenolics Modulates Functional Traits in Lettuce

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1457
Author(s):  
Leilei Zhang ◽  
Erika Martinelli ◽  
Biancamaria Senizza ◽  
Begoña Miras-Moreno ◽  
Evren Yildiztugay ◽  
...  
Keyword(s):  
Functional Traits ◽  
Inhibitory Activity ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Stress Conditions ◽  
Future Application ◽  
Total Phenolic ◽  
Phenolic Profile ◽  
Antioxidant Power ◽  
The Impact

The quest for sustainable strategies aimed at increasing the bioactive properties of plant-based foods has grown quickly. In this work, we investigated the impact of exogenously applied phenolics, i.e., chlorogenic acid (CGA), hesperidin (HES), and their combinations (HES + CGA), on Lactuca sativa L. grown under normal- and mild-salinity conditions. To this aim, the phenolic profile, antioxidant properties, and enzyme inhibitory activity were determined. The untargeted metabolomics profiling revealed that lettuce treated with CGA under non-stressed conditions exhibited the highest total phenolic content (35.98 mg Eq./g). Lettuce samples grown under salt stress showed lower phenolic contents, except for lettuce treated with HES or HES + CGA, when comparing the same treatment between the two conditions. Furthermore, the antioxidant capacity was investigated through DPPH (2,2-diphenyl-1-picrylhydrazyl), ABTS (2,20-azinobis-(3-ethylbenzothiazoline-6-sulfonate)), and FRAP (ferric reducing antioxidant power) assays, coupled with metal-chelating activity and phosphomolybdenum capacity. An exciting increase in radical scavenging capacity was observed in lettuce treated with exogenous phenolics, in both stress and non-stress conditions. The inhibitory activity of the samples was evaluated against target health-related enzymes, namely cholinesterases (acetylcholinesterase; AChE; butyryl cholinesterase; BChE), tyrosinase, α-amylase, and α-glucosidase. Lettuce treated with HES + CGA under non-stress conditions exhibited the strongest inhibition against AChE and BChE, while the same treatment under salinity conditions resulted in the highest inhibition capacity against α-amylase. Additionally, CGA under non-stress conditions exhibited the best inhibitory effect against tyrosinase. All the functional traits investigated were significantly modulated by exogenous phenolics, salinity, and their combination. In more detail, flavonoids, lignans, and stilbenes were the most affected phenolics, whereas glycosidase enzymes and tyrosinase activity were the most affected among enzyme assays. In conclusion, the exogenous application of phenolics to lettuce represents an effective and green strategy to effectively modulate the phenolic profile, antioxidant activity, and enzyme inhibitory effects in lettuce, deserving future application to produce functional plant-based foods in a sustainable way.

scholarly journals In vitro evaluation of the antioxidant, anti-Propioni bacterium acne and antityrosinase effects of Equisetum ramosissimum (Jordanian horsetail)

2020 ◽  
Vol 19 (10) ◽  
pp. 2147-2152
Author(s):  
Noor Sabah Abed Savaya ◽  
Reem Adnan Issa ◽  
Wamidh Hadi Talib
Keyword(s):  
Gallic Acid ◽  
Total Phenolic Content ◽  
Phenolic Content ◽  
Radical Scavenging Activity ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Screening Tests ◽  
Total Phenolic ◽  
Aqueous Methanol

Purpose: The investigation of Equisetum ramosissimum Desf. (Equisetaceae) extracts for total phenolic content, potential antioxidant properties, and antibacterial activity against Propioni bacterium acne and antityrosinase effects.Methods: The aerial parts of the E. ramosissimum from Jordan were extracted by maceration and Soxhlet methods, using solvents of different polarities. The composition of the extracts were qualitatively screened using standard phytochemical tests. Quantitatively, total phenolic content (mg/mL, equivalent to gallic acid), ABTS free radical scavenging activity (IC50), anti-P. acne (MIC and MBC), and tyrosinase inhibitory effects (IC50) were also determined.Results: The aqueous-methanol Soxhlet extract contained the highest total phenolic content (0.675 mg/mL gallic acid equivalents). Besides, phytochemical screening tests revealed the presence of phenols, flavonoids, tannins, alkaloids and saponins in the aqueous methanol Soxhlet extract, contributing to the antioxidant (IC50 = 0.125) and antityrosinase (IC50 = 1.125) effects. This extract also showed potent antimicrobial effects against P. acne (MIC = 6.250 mg/mL; MBC = 12.500 mg/mL). Other extracts, including ethanol, water and ethyl acetate, showed lower total phenolic content with moderate and weak biological activity.Conclusion: E. ramosissimum is a promising plant species to be considered for antioxidant, antiacne, and antityrosinase effects. However, further testing (including invivo, histological examination, and highperformance liquid chromatography (HPLC) analysis) is necessary to understand more about its mechanisms of action. Keywords: Antioxidant, Anti-Propioni bacterium acne, Antityrosinase, Equisetum ramosissimum, Total phenolic content, Jordanian horsetail

scholarly journals The interaction effect and optimal formulation of selected polyherbal extracts towards antioxidant activity

Food Research ◽  
2020 ◽  
Vol 4 (6) ◽  
pp. 2042-2048
Author(s):  
N.F.A. Rahim ◽  
N. Muhammad ◽  
N. Abdullah ◽  
B.A. Talip ◽  
K.H. Poh
Keyword(s):  
Interaction Effect ◽  
Curcuma Longa ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Total Phenolic ◽  
Past Study ◽  
Dpph Assay ◽  
Frap Assay ◽  
Antioxidant Power ◽  
Curry Leaves

Past study showed that lemongrass (Cymbopogon citratus), curry leaves (Murrya koenigii), turmeric (Curcuma longa) and ginger (Zingiber officinale) contain phytochemicals associated with antioxidant properties. However, all the herbs are tested individually and rarely mix together. This study was conducted to examine the antioxidant properties and interaction effect when combined. The plants studied were decocted with distilled water. Eighteen formulations of aqueous extracts were established using simplex lattice mixture design that was generated by Design Expert software. The antioxidant properties were analyzed by 2-diphenyl-2-picryl hydrazyl radical scavenging (DPPH), ferric reducing antioxidant power (FRAP), total phenolic content (TPC) and total flavonoid content (TFC) assays. The result showed that the mixture of lemongrass and curry leaves extracts gave the highest reading in DPPH assay (91.14%), FRAP assay (215.66 mM) and TFC (22.62 µg Rutin/mL). In the DPPH assay, the ratio of one to one (1:1) mixture of lemongrass with other plants extracts showed antagonistic interaction. There were five (5) formulations that showed synergistic interaction in all assays. However, there were two (2) formulations that showed antagonistic interactions on both DPPH and FRAP assays. No additive effect was observed in all formulations. The suggested optimum formulation contains 53.7% lemongrass, 43.4% curry leaves, 2.9% ginger and 0.0% turmeric. Most of the mixtures presented synergistic interactions. This indicated the potential of plant extract mixtures to be developed into nutraceutical products in the future by conducting in-vivo study.

In-Vitro and In-Silico Alpha Glucucosidase Inhibitory activity of Oroxylum indicum

2021 ◽  
pp. 119-125
Author(s):  
Gejalakshmi S. ◽  
Harikrishnan N. ◽  
Anas S. Mohameid
Keyword(s):  
Molecular Docking ◽  
Inhibitory Activity ◽  
In Silico ◽  
Docking Study ◽  
Alpha Amylase ◽  
Scavenging Activity ◽  
Molecular Docking Study ◽  
Ic50 Value ◽  
Alpha Glucosidase

Background: Diabetes mellitus is a metabolic condition characterized by elevated blood glucose levels in the bloodstream. It occurs due to the inadequate amount of insulin secreted in the body or resistance of insulin receptors. Objective: In the present study, for its effect on alpha-amylase and alpha-glucosidase enzymes, Oroxylum indicuma flavone glycoside was assessed using in-vitro assays by removing the respective enzymes from whole wheat and barley in conjunction with in-silico analysis. Method: in-vitro alpha amylase inhibitory activity and in-vitro alpha glucosidase inhibitory activity was performed using acarbose as a standard drug. The molecular docking study was performed using Schrodinger (Maestro V 11.5) software. The parameters glide score, Lipinski rule for drug likeliness, bioactive scoring and ADME properties were assessed in the docking study. In addition, baicalein's antioxidant function was assessed using DPPH assay, nitric oxide scavenging activity. The cytotoxicity of Oroxylum indicumwas evaluated using the Brine shrimp lethality assay. Results: The alpha-amylase assay performed showed IC50 value of 48.40 µg/ml for Oroxylum indicumwhereas alpha-glucosidase assay showed an IC50 value of 16.03 µg/ml. Oroxylum indicumshows the glide score of-5.565 with 5EOF and glide score of -5.339 with 5NN8 in the molecular docking study. The highest percentage of DPPH radical scavenging activity and nitrous oxide scavenging activity were found to be.27% at160 µg/ml and 50.02% at the concentrations of 160 µg/ml respectively. Conclusion: Based on further in vivo and clinical trials, Oroxylum indicummay be used for the management of hyperglycaemia.

Free radical scavenging, Thrombolytic and cytotoxic effects of the medicinal herb, Crassocephalum crepidioides

2021 ◽  
pp. 2205-2210
Author(s):  
Md. Tanvir Kabir ◽  
Mayesha Samiha ◽  
Hasina Yasmin ◽  
Mohammed S. Rahman ◽  
Md. Sohanur Rahman ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Plant Extract ◽  
Radical Scavenging ◽  
Antioxidant Property ◽  
Total Phenolic ◽  
Phytochemical Analysis ◽  
Dpph Assay ◽  
Thrombolytic Activity ◽  
Cytotoxic Effects ◽  
Ic50 Value

Crassocephalum crepidioides belongs to the Asteraceae family and the leaves of the plant were used in the phytochemical screening. The objective of this study was phytochemical analysis of C. crepidioides in order to evaluate its antioxidant, thrombolytic and cytotoxic effects. In this study, DPPH assay and total phenolic content (TPC) were used to evaluate the antioxidant property. Brine shrimp lethality test was performed to find the cytotoxicity. The TPC of methanolic extract of C. crepidioides was 637.22 mg/gm and gallic acid was standard in this procedure. The IC50 value of the plant extract of DPPH assay was 136.016 μg/mL obtained by using ascorbic acid as reference standard. Furthermore, the obtained IC50 value of ascorbic acid was 94.12 μg/mL. In this study, lethal concentrations of vincristine sulfate and the plant extract were 3.064 μg/mL and 69.245 μg/mL respectively. Indeed, the plant exhibited significant antioxidant, cytotoxic and thrombolytic activity.

scholarly journals In Vitro Antioxidant and Antiproliferative Efficacy of Abrus precatorius Seed Extracts On Hep2c Cells and Its Inhibitory Efficacy With Receptors Of Cervical Carcinoma

2021 ◽  
Author(s):  
Amritpal Kaur ◽  
Yash Sharma ◽  
Kamran Waidha ◽  
Madhumita P Ghosh ◽  
Anoop Kumar ◽  
...  
Keyword(s):  
Tannic Acid ◽  
Herbal Remedy ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Seed Extract ◽  
Total Phenolic ◽  
Cervix Carcinoma ◽  
Abrus Precatorius ◽  
Ic50 Value ◽  
Seed Extracts

Abstract Abrus precatorius is a widely distributed tropical medicinal plant with several therapeutic properties, however its seed extract has not been studied against cervical cancer (CaCx) till date. Herein, we have assessed the antioxidant and antiproliferative properties of A. precatorius seed extracts (ethyl acetate and 70% ethanol) prepared from different extraction methods (Soxhlet and maceration) against human cervix carcinoma cells (Hep2C). We observed a significantly higher total flavonoid content of APE (sox) i.e.,112.7±0.127 mg Quercetin Equivalent/g of extract than others; total phenolic content of APA (mac) seed extract was higher i.e.,108.53±0.089 mg Quercetin Equivalent/g of extract and total tannin content of APA (sox) was higher i.e., 98.98± 0.011 mg tannic acid equivalents/g of extract. In addition, tannic acid, rutin and piperine were identified in extracts by HPLC. Furthermore, APA (sox) exhibited the highest radical scavenging activity with an IC50 value of 14.49±0.93µg/ml. APE (mac) showed the most significant antiproliferative activity with IC50 value of 85.90±0.93µg/mL against Hep2c cells. SOD and GST activity was observed as highest in the APA(Mac) extract whereas the catalase activity and GSH content was maximum in the APE(Mac) extract. Further, the MDA content was observed to be the least in APE(Mac) extracts. Docking results suggested maximum binding energy between tannic acid and Her2 receptor compared to doxorubicin (standard). This study provides evidence that A. precatorius seed extracts possess promising bioactive compounds with probable anticancer and antioxidant properties against CaCx which might be utilized as a possible herbal remedy aimed at restricting tumor growth.

scholarly journals Pharmacological Approach to Glycemic Treatment Using Black, Green and Herbal Tea Extracts

2021 ◽  
Vol 50 (3) ◽  
pp. 491-498
Author(s):  
K Akhter ◽  
T Ghous ◽  
Zain ul Abdin ◽  
E Sadaf ◽  
A Hassan ◽  
...  
Keyword(s):  
Green Tea ◽  
Biological Activities ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Black Tea ◽  
In Vitro Assays ◽  
Total Phenolic ◽  
Aqueous Extracts ◽  
Methanolic Extracts ◽  
Alpha Glucosidase

Tea is the second most widely used drink all over the world after water. It has several health benefits. The aqueous and methanolic extracts of black, green and herbal teas for the treatment of hyperglycemia and its associated complications were investigated. Nine tea samples including black, green and herbal teas namely Lipton black tea (LBT), Supreme black tea (SBT), Tapal black tea (TBT), Jasmine green tea (JGT), Lemon grass green tea (LGrT), Lemon green tea (LGT), Berginia ciliata, Bistorta affinis and Paeonia emodi were selected for the study. Total phenolic contents (TPC), total flavonoid contents (TFC), antioxidant properties including inhibitory activity against 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical (ABTS•+), iron chelation and enzymes inhibitory potential for alpha glucosidase and alpha amylase were analyzed based on in vitro assays. The percentage yield of extracts was in the range of 7.2 to 47 %. Methanolic extracts showed greater TPC values (μg GAE/g) while aqueous extracts showed greater TFC values (μg Rutin E/g). The aqueous extracts of most of the studied tea samples showed strong biological activities with very low IC50 (50% inhibitory concentration) values (0.2-69.0 μg/ml) for ABTS•+ radical scavenging, Fe2+ chelation and alpha glucosidase inhibition assays. For amylase inhibition activity, some aqueous extracts including LBT, SBT, JGT and P. emodi exhibited greater IC50 values. Bangladesh J. Bot. 50(3): 491-498, 2021 (September)

scholarly journals Hepatoprotective studies on methanolic extract fractions of Lindernia ciliata and development of qualitative analytical profile for the bioactive extract

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Praneetha Pallerla ◽  
Narsimha Reddy Yellu ◽  
Ravi Kumar Bobbala
Keyword(s):  
Methanolic Extract ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Liver Homogenate ◽  
Reducing Power ◽  
Hepatoprotective Activity ◽  
Total Phenolic ◽  
Effective Fraction

Abstract Background The objective of the study is to evaluate the hepatoprotective activity of methanolic extract fractions of Lindernia ciliata (LC) and development of qualitative analytical profile of the bioactive fraction using HPLC fingerprinting analysis. All the fractions of methanolic extract of Lindernia ciliata (LCME) are assessed for their total phenolic, flavonoid contents and in vitro antioxidant properties by using DPPH, superoxide, nitric oxide, hydroxyl radical scavenging activities and reducing power assay. Acute toxicity study was conducted for all the fractions and the two test doses 50 and 100 mg/kg were selected for the hepatoprotective study. Liver damage was induced in different groups of rats by administering 3 g/kg.b.w.p.o. paracetamol and the effect of fractions were tested for hepatoprotective potential by evaluating serum biochemical parameters and histology of liver of rats. The effective fraction was evaluated for its antihepatotoxic activity against D-Galactosamine (400 mg/kg b.w. i.p.) and in vivo antioxidant parameters viz., Glutathione (GSH), Melondialdehyde (MDA) and Catalase (CAT) levels are estimated using liver homogenate. Results Among all the fractions, butanone fraction of LCME, (BNF-LCME) has shown better hepatoprotective activity and hence it is selected to evaluate the antihepatotoxicity against D-GaIN. The activity of BNF-LCME is well supported in in vitro and in vivo antioxidant studies and may be attributed to flavonoidal, phenolic compounds present in the fraction. Hence, BNF-LCME was subjected to the development of qualitative analytical profile using HPLC finger printing analysis. Conclusions All the fractions of LCME exhibited significant hepatoprotective activity and BNF-LCME (50 mg/kg) was identified as the most effective fraction.

scholarly journals Comparison of the Phytochemical Properties, Antioxidant Activity and Cytotoxic Effect on HepG2 Cells in Mongolian and Taiwanese Rhubarb Species

Molecules ◽  
2021 ◽  
Vol 26 (5) ◽  
pp. 1217
Author(s):  
Ganbolor Jargalsaikhan ◽  
Jin-Yi Wu ◽  
Yen-Chou Chen ◽  
Ling-Ling Yang ◽  
Ming-Shun Wu
Keyword(s):  
Liver Cancer ◽  
Bioactive Compounds ◽  
Cytotoxic Effect ◽  
Antioxidant Properties ◽  
Radical Scavenging ◽  
Ethanol Extract ◽  
Parp Cleavage ◽  
Total Phenolic ◽  
Antioxidant Capacities ◽  
Phytochemical Contents

The Mongolian rhubarb—Rheum undulatum L. (RU)—and Rumex crispus L. (RC)—a Taiwanese local rhubarb belonging to the family of Polygonaceae—are principal therapeutic materials in integrative medicine due to their rich quantities of bioactive compounds; however, their phytochemical and antioxidant properties, and anti-cancer activity is poorly investigated. Furthermore, the phytochemical characteristics of both species may be affected by their different geographical distribution and climatic variance. The current study aimed to compare RU with RC extracts in different polarity solvents (n-hexane, ethyl acetate, acetone, ethanol, and water) for their phytochemical contents including the total phenolic content (TPC), total anthraquinone content (TAC), total flavonoid content (TFC), antioxidant and free radical scavenging capacities, and anticancer ability on the HepG2 cell. Except for the n-hexane extract, all of the RU extracts had considerably higher TPCs than RC extracts, ranging from 8.39 to 11.16 mg gallic acid equivalent (GAE) per gram of dry weight, and the TPCs of each extract were also significantly correlated with their antioxidant capacities by ABTS, DPPH, and FRAP assays (p < 0.05). Moreover, there was no remarkable association between the antioxidant capacities and either TACs or TFCs in both the RU and RC extracts. Besides, high-performance liquid chromatography (HPLC) analysis revealed that both the RU and RC extracts contained chrysophanol, emodin, and physcion, and those bioactive compounds were relatively higher in the n-hexane solvent extracts. Additionally, we observed different levels of dose-dependent cytotoxic effects in all the extracts by cell viability assay. Notably, the ethanol extract of RU had a compelling cytotoxic effect with the lowest half-maximum inhibition concentration (IC50-171.94 ± 6.56 µg/mL at 48 h) among the RU extracts than the ethanol extract of RC. Interestingly, the ethanol extract of RU but not RC significantly induced apoptosis in the human liver cancer cell line, HepG2, with a distinct pattern in caspase-3 activation, resulting in increased PARP cleavage and DNA damage. In summary, Mongolian Rhubarb, RU, showed more phytochemical contents, as well as a higher antioxidant capacity and apoptotic effect to HepG2 than RC; thus, it can be exploited for the proper source of natural antioxidants and liver cancer treatment in further investigation.

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