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The spike (S) protein is a leading vaccine candidate against SARS-CoV-2 infection. The S1 domain of S protein, which contains a critical receptor-binding domain (RBD) antigen, potentially induces protective immunoreactivities against SARS-CoV-2. In this study, we presented preclinical evaluations of a novel insect cell-derived SARS-CoV-2 recombinant S1 (rS1) protein as a potent COVID-19 vaccine candidate. The native antigenicity of rS1 was characterized by enzyme-linked immunosorbent assay with a neutralizing monoclonal antibody targeting the RBD antigen. To improve its immunogenicity, rS1-adjuvanted with fucoidan/trimethylchitosan nanoparticles (FUC-TMC NPs) and cytosine-phosphate-guanosine-oligodeoxynucleotides (CpG-ODNs) were investigated using a mouse model. The S1-specific immunoglobulin G (IgG) titers, FluoroSpot assay, pseudovirus- and prototype SARS-CoV-2-based neutralization assays were assessed. The results showed that the rS1/CpG/ FUC-TMC NPs (rS1/CpG/NPs) formulation induced a broad-spectrum IgG response with potent, long-lasting, and cross-protective neutralizing activity against the emerging SARS-CoV-2 variant of concern, along with a Th1-biased cellular response. Thus, the rS1/CpG/NPs formulation presents a promising vaccination approach against COVID-19.

Infections caused by extensively drug-resistant (XDR) Acinetobacter nosocomialis have become a challenging problem. The frequent use of colistin as the last resort drug for XDR bacteria has led to the emergence of colistin-resistant A. nosocomialis (ColRAN) in hospitals. The mechanism of colistin resistance in A. nosocomialis remains unclear. This study aimed to investigate the mechanisms underlying colistin resistance in clinical ColRAN isolates. We collected 36 A. nosocomialis isolates from clinical blood cultures, including 24 ColRAN and 12 colistin-susceptible A. nosocomialis (ColSAN). The 24 ColRAN isolates clustered with ST1272 (13), ST433 (eight), ST1275 (two), and ST410 (one) by multilocus sequence typing. There was a positive relationship between pmrCAB operon expression and colistin resistance. Further analysis showed that colistin resistance was related to an amino acid substitution, Ser253Leu in PmrB. By introducing a series of recombinant PmrB constructs into a PmrB knockout strain and protein structural model analyses, we demonstrated that the association between Ser253Leu and Leu244 in PmrB was coupled with colistin resistance in ColRAN. To the best of our knowledge, this is the first study demonstrating that the key amino acid Ser253Leu in PmrB is associated with overexpression of the pmrCAB operon and hence colistin resistance. This study provides insight into the mechanism of colistin resistance in A. nosocomialis.

The ongoing COVID-19 (i.e., coronavirus) pandemic continues to adversely affect the human life, economy, and the world's ecosystem. Although significant progress has been made in developing antiviral materials for the coronavirus, much more work is still needed. In this work, N-functionalized graphene quantum dots (GQDs) were designed and synthesized as the antiviral nanomaterial for Feline Coronavirus NTU156 (FCoV NTU156) and Enterovirus 71 (EV71)) with ultra-high inhibition (99.9%). To prepare the GQD samples, a unique solid-phase microwave-assisted technique was developed and the cell toxicity was established on the H171 and H184 cell lines after 72 h incubation, indicating superior biocompatibility. The surface functionality of GQDs (i.e., the phenolic and amino groups) plays a vital role in interacting with the receptor-binding-domain of the spike protein. It was also found that the addition of polyethylene glycol is advantageous for the dispersion and the adsorption of functionalized GQDs onto the virus surface, leading to an enhanced virus inhibition. The functionality of as-prepared GQD nanomaterials was further confirmed where a functionalized GQD-coated glass was shown to be extremely effective in hindering the virus spread for a relatively long period (20 h).

Backgrounds While severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection presents with mild or no symptoms in most cases, a significant number of patients become critically ill. Remdesivir has been approved for the treatment of coronavirus disease 2019 (COVID-19) in several countries, but its use as monotherapy has not substantially lowered mortality rates. Because agents from traditional Chinese medicine (TCM) have been successfully utilized to treat pandemic and endemic diseases, we designed the current study to identify novel anti-SARS-CoV-2 agents from TCM. Materials and methods We initially used an antivirus-induced cell death assay to screen a panel of herbal extracts. The inhibition of the viral infection step was investigated through a time-of-drug-addition assay, whereas a plaque reduction assay was carried out to validate the antiviral activity. Direct interaction of the candidate TCM compound with viral particles was assessed using a viral inactivation assay. Finally, the potential synergistic efficacy of remdesivir and the TCM compound was examined with a combination assay. Results The herbal medicine Perilla leaf extract (PLE, approval number 022427 issued by the Department of Health and Welfare of Taiwan) had EC50 of 0.12 ± 0.06 mg/mL against SARS-CoV-2 in Vero E6 cells – with a selectivity index of 40.65. Non-cytotoxic PLE concentrations were capable of blocking viral RNA and protein synthesis. In addition, they significantly decreased virus-induced cytokine release and viral protein/RNA levels in the human lung epithelial cell line Calu-3. PLE inhibited viral replication by inactivating the virion and showed additive-to-synergistic efficacy against SARS-CoV-2 when used in combination with remdesivir. Conclusions Our results demonstrate for the first time that PLE is capable of inhibiting SARS-CoV-2 replication by inactivating the virion. Our data may prompt additional investigation on the clinical usefulness of PLE for preventing or treating COVID-19., Graphical abstract Image 1

Nowadays, chronic kidney disease (CKD) becomes a principal barrier in clinical diagnosis and treatment. For clinical patients, chronic kidney disease will potentially lead to multiple lesion that increase the risk of mortality. The most significant challenge in CKD is detecting uremic toxins, including small water-soluble solutes (uric acid, urea and creatinine) and protein-bound solutes (p-cresol and indoxyl sulfate). Surface-enhanced Raman scattering (SERS) platform is rapid and sensitive nanotechnology for bio-detection. Thus, the floating-typed SERS substrate is prepared by embedded silver nanoparticles (AgNPs) on the poly (diallyldimethyl-ammonium) chloride (PDDA) modified graphene oxide (GO) nanosheets for the biomolecules and uremic toxins detection. The optimal interparticle distances of AgNPs are modulated to generate the strong “hot spots” for enhancing Raman signals. The characterizations of AgNPs/GO-PDDA nanosheets are evaluated by transmission electron microscopy, zeta potential, FTIR, X-ray photoelectron spectroscopy, and Raman spectroscopy. The results show that the floating SERS-active substrate provides quantitatively linear measurement (i.e., 3.8 × 10−2 ˜10-5 M of urea) and ultrasensitive SERS detection (i.e., detection limit of adenine: lower than 10-10 M) of biomolecules, which offers great potential for practical clinical applications in rapid and label-free detection of clinical uremic toxins.

Grail is a well-characterized mediator of metabolic disease, tumour progression, and immune response. However, its role in influenza A virus (IAV) infection remains poorly understood. In this study, we demonstrated that Grail knockdown potentiates IAV infection, whereas Grail overexpression blocks IAV replication. The intranasal administration of IAV to Grail KO mice led to a lower survival rate than in similarly infected wild-type mice. Additionally, IAV-infected Grail KO mice had higher viral titres, greater immune cell infiltration, and increased expression of inflammatory cytokines in the lungs. Mechanistically, we showed that Grail interacts with viral nucleoprotein (NP), targeting it for degradation and inhibiting IAV replication. NP expression was increased in Grail knockdown cells and reduced in cells overexpressing Grail. Collectively, our results demonstrate that Grail acts as a negative regulator of IAV infection and replication by degrading viral NP. These data increase our understanding of the host antiviral response to infection with IAV.

In obese adults, nonalcoholic fatty liver disease (NAFLD) is accompanied by multiple metabolic dysfunctions. Although upregulated hepatic fatty acid synthesis has been identified as a crucial mediator of NAFLD development, the underlying mechanisms are yet to be elucidated. In this study, we reported upregulated expression of gene related to anergy in lymphocytes (GRAIL) in the livers of humans and mice with hepatic steatosis. Grail ablation markedly alleviated the high-fat diet-induced hepatic fat accumulation and expression of genes related to the lipid metabolism, in vitro and in vivo. Conversely, overexpression of GRAIL exacerbated lipid accumulation and enhanced the expression of lipid metabolic genes in mice and liver cells. Our results demonstrated that Grail regulated the lipid accumulation in hepatic steatosis via interaction with sirtuin 1. Thus, Grail poses as a significant molecular regulator in the development of NAFLD.

Grail is a crucial regulator of various biological processes, including the development of T-cell anergy, antiviral innate immune response, and cancer. However, the role of Grail in adipogenesis and obesity remains unclear. Here, we demonstrated that Grail knockdown in vitro leads to a decrease in PPARγ expression, resulting in adipogenesis inhibition. However, Grail overexpression induced the same effects. Grail was shown to interact with PPARγ, targeting it for degradation and modulating its adipogenic activity. PPARγ expression was shown to be considerably reduced in Grail knockout (KO) mice fed normal diet or high-fat diet (HFD). The administration of both normal diet or HFD to Grail KO mice led to lower adipose mass and body weight than those in the wild-type mice. HFD-fed Grail KO mice had improved glucose and insulin tolerance. Taken together, our results indicate that Grail plays a pivotal role in adipogenesis and diet-induced obesity by regulating PPARγ activity.

We report an extremely sensitive and ultra-low-cost PVDF-based SERS substrate using an easy fabrication process that can be conducted in a laboratory and scaled up for industrial applications. Hydrophilic PVDF membranes were sequentially dipped in ethanol and aqueous sodium chloride solution, following which multi-shaped Ag nanoparticles were deposited on the surface of the membranes by suction filtration. The simple solvating pretreatment led to an approximately 188-fold enhancement in the SERS signal intensity of R6G. Using the hydrophilic PVDF-based SERS substrate, subnanomolar sensitivity for R6G was achieved using 3 μL of sample volume on a 3×3 mm2 SERS substrate within 1 min detection time and without the need for concentration or drying processes. To demonstrate the feasibility of the developed SERS substrate as a sensor for bacterial spore detection, it was applied for the detection of the representative biomarker dipicolinic acid (DPA). The limits of detection of DPA and Bacillus spores were measured as 1 ppm and 5×103 spores/mL, respectively. The optimized configuration of low substrate area and wet micro-analyte volume makes the PVDF-based SERS substrate a simple and low-cost tool for the trace-level detection of small molecules in a rapid, sensitive, and high-throughput manner.

Fucoidan/trimethylchitosan nanoparticles (FUC-TMC-NPs) have the potential to improve the immunostimulating efficiency of anthrax vaccine adsorbed (AVA). FUC-TMC-NPs with positive (+) or negative (–) surface charges were prepared via polyelectrolyte complexation, both charged NP types permitted high viability and presented no cytotoxicity on L929, A549 and JAWS II dendritic cells. Flow cytometry measurements indicated lower (+)-FUC-TMC-NPs internalization levels than (–)-FUC-TMC-NPs, yet produced high levels of pro-inflammatory cytokines IFN-γ, IL12p40, and IL-4. Moreover, fluorescence microscope images proved that both charged NP could deliver drugs into the nucleus. In vivo studies on A/J mice showed that (+)-FUC-TMC-NPs carrying AVA triggered an efficient response with a higher IgG anti-PA antibody titer than AVA with CpG oligodeoxynucleotides, and yielded 100 % protection when challenged with the anthracis spores. Furthermore, PA-specific IgG1 and IgG2a analysis confirmed that (+)-FUC-TMC-NPs strongly stimulated humoral immunity. In conclusion, (+)-FUC-TMC-NP is promising anthrax vaccine adjuvant as an alternative to CpG.

We examined the efficacy of fucoidan-N-(2-hydroxy-3-trimethylammonium)propylchitosan nanoparticles (FUC-HTCC NPs) as adjuvants for anthrax vaccine adsorbed (AVA). Positively and negatively surface-charged FUC-HTCC NPs were prepared via polyelectrolyte complexation by varying the mass ratio of FUC and HTCC. When cultured with L929 cells or JAWS II dendritic cells, both charged NPs showed high cell viability and low cytotoxicity, observed via MTT assay and lactate dehydrogenase release assay, respectively. In addition, we have monitored excellent NPs uptake efficacy by dendritic cells and observed that combining FUC-HTCC NPs with AVA significantly increases the magnitude of IgG-anti-protective antigen titers in A/J mice compared to that by CpG oligodeoxynucleotides plus AVA or AVA alone, and PA-specific IgG1 and IgG2a analysis confirmed that FUC-HTCC NPs strongly stimulated humoral immunity. Furthermore, FUC-HTCC NPs plus AVA provided a superior survival rate (100%) of A/J mice compared to CpG oligodeoxynucleotides plus AVA (75%) or AVA alone (50%) following anthrax lethal toxin challenge. The findings support FUC-HTCC NPs as a potential adjuvant of AVA for rapid induction of protective immunity.

Herein, positively surface-charged silver nanoparticles (AgNPs) capped with trimethylchitosan nitrate (TMCN) were synthesized using an environmentally friendly method. Nano-sized TMCN-AgNPs (~80 nm) with high zeta potential (>30 mV) provide sufficient static repulsion to stabilize colloid AgNPs in aqueous solutions without aggregation for >3 months. In in vitro cell cycle assays, TMCN-AgNPs showed low cytotoxicity towards L929 cells. A microdilution inhibition assay demonstrated the antifungal potential of TMCN-AgNPs, with a minimum inhibitory concentration of 0.06 mM against Candida tropicalis ATCC 750, and 0.46 mM against both Candida albicans ATCC 76615 and Candida glabrata ATCC 15545. Moreover, the addition of TMCN-AgNPs at 0.23 mM significantly reduced biofilm formation in 96-well plates with C. albicans and C. tropicalis. Importantly, when zebrafish eggs were infected with Candida cells, 0.23 mM TMCN-AgNPs greatly diminished the amount of biofilm on eggs and rescued the survival of embryos by up to 70%.

A facile polyelectrolyte complexation method for the preparation of both positively and negatively surface charged nanoparticles composed of chondroitin sulfate (ChS) and N-[(2-hydroxy-3-trimethylammonium)propyl]chitosan (HTCC) is reported. Production of ChS-HTCC nanoparticles with reverse zeta potential was easily controlled by varying the ChS/HTCC mass ratio. The encapsulation efficiency increased with the increase in initial FITC-BSA concentration in positively charged NPs and reached 75%. However, a maximum of 20% encapsulation efficiency was achieved in the case of negatively charged NPs. In vitro release studies of positively charged ChS-HTCC NPs showed a small burst effect followed by a continued and controlled release. Both charges of ChS-HTCC NPs showed no cytotoxicity in HUVECs. The confocal images showed that ChS-HTCC NPs of both charges can be incorporated and retained by the A549 cells. Flow cytometric analysis data demonstrated that ChS-HTCC NPs of both charges were detected in more than 80% of the A549 cells.

Herein, we describe an improved procedure for the green synthesis of chondroitin sulfate stabilized silver nanoparticles (ChS-AgNPs). Glucose was used as a reducing agent under alkaline conditions to obtain a small particle size (10 nm), and the reduction was complete within one hour at room temperature. The concentration of NaOH affected the reaction rate, formation yield, and particle size of ChS-AgNPs. The formation of AgNPs was confirmed using UV-vis, TEM, XRD, and XPS. ChS-AgNPs showed excellent catalytic activities in the reduction of 4-nitrophenol by NaBH

In this study, we synthesized various quaternary chitosan derivatives and used them to stabilize gold nanoparticles (AuNPs). These chitosan derivatives comprised N-(2-hydroxy)propyl-3-trimethylammonium chitosan chloride (HTCC), folate-HTCC, galactosyl-HTCC, and their fluorescein isothiocyanate-conjugated derivatives. Various positively surface-charged AuNPs were prepared under alkaline conditions using glucose as a reducing agent in the presence of the HTCC derivatives (HTCCs). The effects of the concentration of NaOH, glucose, and HTCCs on the particles size, zeta potential, and stability were studied in detail. Cell cycle assays verify that none of the HTCCs or HTCCs-AuNPs was cytotoxic to human umbilical vein endothelial cells. Flow cytometry analysis showed that the folate HTCC-AuNPs were internalized in Caco-2, HepG2, and HeLa cancer cells to a significantly greater extent than AuNPs without folate. But, galactosyl HTCC-AuNPs only showed high cell uptake by HepG2 cells.

We report a facile route for the green synthesis of trimethylchitosan nitrate-capped silver nanoparticles (TMCN-AgNPs) with positive surface charge. In this synthesis, silver nitrate, glucose, and trimethyl chitosan nitrate (TMCN) were used as silver precursor, reducing agent, and stabilizer, respectively. The reaction was carried out in a stirred basic aqueous medium at room temperature without the use of energy-consuming or expensive equipment. We investigated the effects of the concentrations of NaOH, glucose, and TMCN on the particle size, zeta potential, and formation yield. The AgNPs were characterized by UV-visible spectroscopy, photon correlation spectroscopy, laser Doppler anemometry, transmission electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The catalytic activity of the TMCN-AgNPs was studied by the reduction of 4-nitrophenol using NaBH4 as a reducing agent. We evaluated the antibacterial effects of the TMCN-AgNPs on Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus using the broth microdilution method. The results showed that both gram-positive and gram-negative bacteria were killed by the TMCN-AgNPs at very low concentration (< 6.13 μg/mL). Moreover, the TMCN-AgNPs also showed high antibacterial activity against clinically isolated multidrug-resistant A. baumannii strains, and the minimum inhibitory concentration (MIC) was ≤ 12.25 μg/mL.

Background and Aim: Botulinum neurotoxin Type E (BoNT/E), one of the most lethal toxin known, is the common contamination in fishery products or fish consumption that causes foodborne botulism. It is necessary to establish a sensitive and specific method for the detection of BoNT/E because of its extremely low lethal dose. Methods: In this study, a practical enzyme-linked immunosorbent assay for BoNT/E detection was developed. The assay is based on the sandwich format using monoclonal antibodies of two distinct specificities. An affinity-purified anti-BoNT/E light chain Mab (1E1) is utilized to adsorb BoNT/E from solution, and the second anti-BoNT/E heavy chain C terminus Mab (5E1) conjugated with peroxidase is then used to form sandwich complexes, and peroxidase allows color development and measurement of optical density at 450 nm. Results: Standard curves were linear over the range of 5–100 ng/ml BoNT/E. The limit of detection was below 10 ng/ml in phosphate-buffered saline buffer. The developed BoNT/E assay also showed no cross-reaction to Type A neurotoxin (BoNT/A) and Type B neurotoxin (BoNT/B). Conclusion: Herein, a sensitive and accurate ELISA for BoNT/E detection was presented. It has the potential to utilize in vivo BoNT/E analysis and contamination monitoring.

Positively and negatively surface-charged nanoparticles (NPs) were prepared with chondroitin sulfate (ChS) and trimethylchitosan (TMC). NP size, surface charge, formation yield, and water content were investigated as a function of weight ratio and concentration. Size and zeta potential were controlled by varying the ChS/TMC mass ratio. FTIR spectra revealed interactions among composite NP constituents. TEM images showed that the NPs were nearly spherical, with an average size of ∼ 300 nm. Encapsulation efficiency increased in positively charged NPs with increases in fluorescein isothiocyanate-bovine serum albumin concentration. Negatively charged NPs had only 10-20% encapsulation efficiency. The release profile, release kinetics and mechanism of positively charged ChS-TMC NPs were studied in vitro. NP cytocompatibility and uptake were verified ex vivo. Both types of NPs were taken up and retained in cells. A549 cells took up more positively charged (49.4%) than negatively charged (35.5%) NPs.

Primary sternal osteomyelitis (PSO) is a rare syndrome. In adults, it usually occurs with underlying predisposing factors, such as immunodeficiency, or intravenous (IV) drug abuse. The infecting organism in these patients is usually Staphylococcus aureus or Pseudomonas aeruginosa. Peptostreptococcus species are Gram-positive anaerobic cocci and are part of the normal flora of human mucocutaneous surfaces. Peptostreptococcus infection can occur in all body sites, including the central nervous system, head, neck, chest, abdomen, pelvis, skin, bone, joint, and soft tissue. Here, we report on a 32-year-old previously healthy Chinese man who was diagnosed with PSO and P. anaerobius was yielded in the bacterial culture. He was treated empirically with antibiotics, but these failed. After additional limited surgical intervention with debridement, the PSO was cured.

A one-step route for the green synthesis of highly stable and nanosized silver metal particles with narrow distribution is reported. In this environmentally friendly synthetic method, silver nitrate was used as silver precursor and biocompatible chondroitin sulfate (ChS) was used as both reducing agent and stabilizing agent. The reaction was carried out in a stirring aqueous medium at the room temperature without any assisted by microwave, autoclave, laser irradiation, γ-ray irradiation or UV irradiation. The transparent colorless solution was converted to the characteristics light red then deep red-brown color as the reaction proceeds, indicating the formation of silver nanoparticles (Ag NPs). The Ag NPs were characterized by UV–visible spectroscopy (UV–vis), photon correlation spectroscopy, laser Doppler anemometry, transmission electron microscopy (TEM), and Fourier-transform infrared spectroscopy (FT-IR). The results demonstrated that the obtained metallic nanoparticles were Ag NPs capped with ChS. In this report, dynamic light scattering (DLS) was used as a routinely analytical tool for measuring size and distribution in a liquid environment. The effects of the reaction time, reaction temperature, concentration and the weight ratio of ChS/Ag+ on the particle size and zeta potential were investigated. The TEM image clearly shows the morphology of the well-dispersed ChS-capped Ag NPs are spherical in shape, and the average size (

Dar-Shih Hsieh1,2, Hsiu-Chin Lu5, Cheng-Cheung Chen1,3, Chang-Jer Wu1,*, Ming-Kung Yeh4,* 1Department of Food Science, National Taiwan Ocean University, Keelung, Taiwan; 2Division of Urology, 3Department of Pathology, Tri-Service General Hospital, 4Institute of Preventive Medicine, National Defence Medical Center, Taipei, Taiwan; 5Department of Laboratory Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan*These authors contributed equally to this workAbstract: Nanogold particles are commonly used in nanomedicine. We generated physical nanogold (pNG) conjugated with different ratios of epigallocatechin-3-gallate (EGCG) and evaluated its physicochemical properties, antioxidant activity, and cytotoxicity in vitro as well as anticancer activity in vivo. Results showed that the EGCG-pNG conjugates were successfully prepared at ratios between 23:1 and 23:5, with the percentage of EGCG content increasing with the EGCG:pNG ratio from 23:1 (2.0% ± 0.02%) to 23:5 (28% ± 0.3%). EGCG-pNG particles at ratios of 23:1 and 23:5 demonstrated significantly decreased size from 500 to 20 nm and decreasing zeta potentials of 21 mV to –22 mV, respectively. At a ratio of 23:2.5, the EGCG-pNG particles (27% EGCG, 50 nm in size, zeta potential of –8 mV) showed longer EGCG activity half-life (110 days vs 5 hours), controlled release (2 hours vs 30 minutes), and higher antioxidant activity (four times), as well as inhibition of tumor cell growth, than controls. The present study indicated that EGCG-pNG possesses promising therapeutic potential, based on its strong free-radical scavenging and anticancer activities.Keywords: EGCG, nanoparticles, antioxidant, antitumor activity, nanogold

Cheng-Cheung Chen,1,2 Dar-Shih Hsieh,1,3 Kao-Jean Huang,4 Yi-Lin Chan,5 Po-Da Hong,6 Ming-Kung Yeh,6–8,* Chang-Jer Wu1,*1Department of Food Science, National Taiwan Ocean University, Keelung, 2Institute of Preventive Medicine, National Defense Medical Center, Taipei, 3Division of Urology, Department of Surgery, Ren-Ai Hospital, Shulin, New Taipei City, 4Department of Life Science and Institute of Biotechnology, National Dong Hwa University, Hualien, 5Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, 6Materials Technology Program, Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei, 7School of Pharmacy, National Defense Medical Center, Taipei, 8Food and Drug Administration, Ministry of Health and Welfare, Taipei, Taiwan, Republic of China*These authors contributed equally to this workAbstract: (-)-Epigallocatechin-3-gallate (EGCG), the major bioactive constituent in green tea, has been reported to effectively inhibit the formation and development of tumors. To maximize the effectiveness of EGCG, we attached it to nanogold particles (EGCG-pNG) in various ratios to examine in vitro cytotoxicity and in vivo anti-cancer activity. EGCG-pNG showed improved anti-cancer efficacy in B16F10 murine melanoma cells; the cytotoxic effect in the melanoma cells treated with EGCG-pNG was 4.91 times higher than those treated with EGCG. The enhancement is achieved through mitochondrial pathway-mediated apoptosis as determined by annexin V assay, JC-10 staining, and caspase-3, -8, -9 activity assay. Moreover, EGCG-pNG was 1.66 times more potent than EGCG for inhibition of tumor growth in a murine melanoma model. In the hemolysis assay, the pNG surface conjugated with EGCG is most likely the key factor that contributes to the decreased release of hemoglobin from human red blood cells.Keywords: gold nanoparticles, EGCG, anticancer, melanoma

Cancer is a major cause of deaths. More recently, drug delivery systems (DDSs) using gold nanoparticles (GNPs) have been widely developed in cancer therapy for their easily synthesised, functionalised, better biocompatible, and low toxicity properties. Gold nanoparticle–based platforms as cancer–targeted molecules delivery to tumour tissues have been proposed as an effective strategy by overcoming various biological barriers and increasing retention effect. Coupled with the active targeting on the specific sites during cancer process they can facilitate anticancer effects. This review is focused on the current applications of GNPs as improving cancer–targeted delivery platforms.