Enhancing Temozolomide (TMZ) chemosensitivity using CRISPR-dCas9-mediated downregulation of O6-methylguanine DNA methyltransferase (MGMT).

PURPOSE: Glioblastoma (GBM) stands out as the most prevalent and aggressive intracranial tumor, notorious for its poor prognosis. The current standard-of-care for GBM patients involves surgical resection followed by radiotherapy, combined with concurrent and adjuvant chemotherapy using Temozolomide (TMZ). The effectiveness of TMZ primarily relies on the activity of O6-methylguanine DNA methyltransferase (MGMT), which removes alkyl adducts from the O6 position of guanine at the DNA level, thereby counteracting the toxic effects of TMZ. METHOD: In this study, we employed fusions of catalytically-inactive Cas9 (dCas9) to DNA methyltransferases (dCas9-DNMT3A) to selectively downregulation MGMT transcription by inducing methylation at MGMT promoter and K-M enhancer. RESULT: Our findings demonstrate a significant reduction in MGMT expression, leading to intensified TMZ sensitivity in the HEK293T cell line. CONCLUSION: This study serves as a proof of concept for the utilization of CRISPR-based gene suppression to overcome TMZ resistance and enhance the lethal effect of TMZ in glioblastoma tumor cells.

Development of a highly porous bioscaffold by the combination of bubble entrapping and freezing-thawing techniques to fabricate hyaluronic acid/gelatin tri-layer wound dressing.

The electrospun nanofibers and porous scaffolds hold great promise in regenerative medicine. A novel nanofiber-hydrogel­silicone tri-layer wound dressing has been designed and fabricated to address the limitations of each platform. The bottom nanofiber layer with a 110 mm diameter meets the wound surface and regulates cell attachment and migration. The middle hydrogel layer was fabricated through the optimization of chemical crosslink formation and freezing-thawing cycles (physical crosslink). The fabricated hydrogel with interconnected porous structure has optimized properties (gel fraction (89.45 %) and porosity (80 %)) for wound dressing application. The silicone layer on the outer surface was designed to fix the wound dressing on the skin and prevent the penetration of pathogens. The scanning electron microscope micrograph showed structural integrity in the tri-layer scaffold. In vivo data showed that the tri-layer scaffold accelerates wound healing in the mice model and angiogenesis in the chorioallantoic membrane model. Therefore, the designed scaffold inspired by the skin's structure can be used as a wound dressing to treat wounds.

Synthesis of optimized propolis solid lipid nanoparticles with desirable antimicrobial, antioxidant, and anti-cancer properties.

This study aimed to produce stable propolis nanoparticles with a size below 100 nm, suitable for various applications in industries such as pharmaceuticals, medicine, cosmetics, food, and packaging. To achieve this, propolis solid lipid nanoparticles (PSLNs) were synthesized using the hot homogenization method, and the optimized nanoparticles were analyzed using Design Expert software. The properties of the synthesized PSLN were characterized using UV-visible spectroscopy, FTIR, XRD, PSA, TEM, and zeta potential analysis. The results indicated that PSLNs with a size range of 57 ± 15 nm remained stable in an aqueous medium at pH 7.4. HPLC analysis showed that the active ingredient of phenols and flavonoids in the extract remained stable after the formation of PSLNs. Antioxidant and antibacterial properties of the extract and nanoparticles were also evaluated. The results demonstrated that the biological properties of the extract were effectively preserved in PSLNs, Additionally, the PSLN synthesized exhibited remarkable anticancer properties against the A549 cell line and with IC50 of 0.01 mg/ml after 72 h-treatment. In conclusion, the optimized PSLNs can be utilized as antioxidant and antibacterial additives and have the potential to be used as a drug or drug carrier for the treatment of lung cancer.

Comparison of different methods for synthesis of iron oxide nanoparticles and investigation of their cellular properties, and antioxidant potential.

Iron oxide nanoparticles could play a useful role in lung cancer therapy. Iron oxide nanoparticles (NPs) were synthesized by plant mediated synthesis, chemical, and microbial mediated synthesis. iron oxide nanoparticle polyethylene glycol cis-diamminedichloroplatinum (Fe2O3@PEG@CDDP(, iron oxide nanoparticle polyethylene glycol (Fe2O3@PEG), and cis-diamminedichloroplatinum (CDDP) were evaluated for their antioxidant,and in vitro cytotoxicity tests. X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Field emission scanning electron microscopy (FE-SEM), mapping, and zeta potential were used to characterize the synthesized iron oxides NPs. Cell toxicity was determined using A549 and HFF cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The antioxidant scavenging activity of Fe2O3@PEG@CDDP, Fe2O3@PEG, and CDDP displayed IC50 values (11.96, 26.74, and 3.17 µg/ml) and (8.54, 11.4, and 1.14 µg/ml) in 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays, respectively. Nanoparticles obtained from plant mediated synthesis method showed the great antioxidant activity. Results showed that, green-method synthesized nanoparticles were the most effective at killing cancer cells. Thus, the characteristics of nanoparticles from green synthesis are more valuable than the other methods. Green synthesis is environmental friendly cost-effective, and easy approach for synthesize NPs.

Co-administration of curcumin with other phytochemicals improves anticancer activity by regulating multiple molecular targets.

Natural plant phytochemicals are effective against different types of diseases, including cancer. Curcumin, a powerful herbal polyphenol, exerts inhibitory effects on cancer cell proliferation, angiogenesis, invasion, and metastasis through interaction with different molecular targets. However, the clinical use of curcumin is limited due to poor solubility in water and metabolism in the liver and intestine. The synergistic effects of curcumin with some phytochemicals such as resveratrol, quercetin, epigallocatechin-3-gallate, and piperine can improve its clinical efficacy in cancer treatment. The present review specifically focuses on anticancer mechanisms related to the co-administration of curcumin with other phytochemicals, including resveratrol, quercetin, epigallocatechin-3-gallate, and piperine. According to the molecular evidence, the phytochemical combinations exert synergistic effects on suppressing cell proliferation, reducing cellular invasion, and inducing apoptosis and cell cycle arrest. This review also emphasizes the significance of the co-delivery vehicles-based nanoparticles of such bioactive phytochemicals that could improve their bioavailability and reduce their systemic dose. Further high-quality studies are needed to firmly establish the clinical efficacy of the phytochemical combinations.

AR13 peptide-conjugated liposomes improve the antitumor efficacy of doxorubicin in mice bearing C26 colon carcinoma; in silico, in vitro, and in vivo study.

Currently, liposomes have emerged as efficient and safer nano-carriers for targeted therapy in different cancers. This work aimed to employ PEGylated liposomal doxorubicin (Doxil®/PLD), modified with AR13 peptide, to target Muc1 on the surface of colon cancerous cells. We performed molecular docking and simulation studies (using Gromacs package) of AR13 peptide against Muc1 to analyze and visualize the peptide-Muc1 binding combination. For in vitro analysis, the AR13 peptide was post-inserted into Doxil® and verified by TLC, 1H NMR, and HPLC techniques. The zeta potential, TEM, release, cell uptake, competition assay, and cytotoxicity studies were performed. In vivo antitumor activities and survival analysis on mice bearing C26 colon carcinoma were studied. Results showed that after 100 ns simulation, a stable complex between AR13 and Muc1 formed, and molecular dynamics analysis confirmed this interaction. In vitro analysis demonstrated significant enhancement of cellular binding and cell uptake. The results of in vivo study on BALB/c mice bearing C26 colon carcinoma, revealed an extended survival time to 44 days and higher tumor growth inhibition compared to Doxil®. Thus, the AR13 peptide could be explored as a potent ligand for Muc1, improving therapeutic antitumor efficiency in colon cancer cells.

Cross-Linked Electrospun pH-Sensitive Nanofibers Adsorbed with Temporin-Ra for Promoting Wound Healing.

Bioresponsive nanodrug delivery systems have excellent potential in tissue engineering applications. Poly-anionic and poly-cationic biopolymers have provided a superior platform for designing pH-sensitive drug delivery systems. In this regard, hyaluronic acid-chitosan-polyvinyl alcohol complex nanofibers with high quality and reproducibility were produced by optimizing the solution preparation process. In addition, the synthesized composite nanofiber, with 66.82 kN/mm toughness, 200% swelling ratio, and 60% porosity, exhibited excellent properties to meet the requirements of the ideal wound dressing. Green cross-linking with citric acid prevented the destruction of the nanofiber even after prolonged immersion in biological solutions. ζ potential studies demonstrated that the synthesized nanofiber has a negative surface charge (∼-30) at physiological pH. The pKa of the temporin-Ra peptide is about 10, and as a result the peptide molecules have a net positive charge in physiological conditions. Therefore, peptide molecules immobilized on the synthesized scaffold based on surface adsorption. In vivo evaluation has proven that the wound bed has an alkaline environment, facilitating peptide release from the nanofiber scaffold. Electrospun nanofibers can imitate the architecture of the extracellular matrix for accelerating wound healing. In vitro investigation showed better adhesion, proliferation, migration, and fibroblast cell growth on peptide-loaded nanofiber samples than other groups. In vivo studies on full-thickness wounds in the mouse model indicated that the designed nanofiber was gradually absorbed without causing dryness or infection. On day 6, the peptide-loaded nanofiber revealed 60% wound closure compared to the control group (17%). In addition, based on histological studies, the composite nanofiber demonstrated excellent tissue repair ability, hence these active nanofiber mats can be a good alternative to existing wound dressings. Gene expression studies show that the antimicrobial peptide promotes the inflammatory phase of wound healing in a shorter time frame by accelerating the tumor necrosis factor-α cytokine response.

Preparation of chitosan nanoparticle containing recombinant CD44v antigen and evaluation of its immunization capacity against breast cancer in BALB/c mice.

OBJECTIVE(S): Breast tumors show heterogeneity containing cancer stem cells as a small subpopulation of a tumor mass. CD44 as a cancer stem cells antigen is abnormally expressed by carcinomas of epithelial origin. Also, overexpression of CD44 variable isoforms (CD44v) is associated with malignancy in breast cancer. In the present research, our objective was to evaluate the immunogenicity of prepared nanoparticles containing a novel recombinant CD44v (rCD44v) protein in the mouse model. MATERIALS AND METHODS: CD44 gene was expressed in E. coli BL21 DE3 using the pET28a-CD44 vector. The expressed rCD44v protein was purified, encapsulated into the chitosan nanoparticles, and administered to BALB/c mice. ELISA was used to evaluate the immunoglobulin levels of immunized animals. For challenge experiment, 2 × 106 4T1-CD44 tumor cells were injected subcutaneously in mice, and tumor size, necrosis, and metastases were measured. Finally, cell proliferation assay, cytokines assay, and neutralization assay of the mouse anti-rCD44v on the human breast cancer cell line were examined. RESULTS: The measured size of chitosan-rCD44v nanoparticles was 146.5 nm. Recombinant CD44v encapsulated by chitosan nanoparticles increases immunological responses via the adjuvant nature of chitosan nanoparticles. In the immunized mice, IgG and IgA titers were significantly increased. Tumor growth in injection and nano-injection test groups compared with the mice control groups displayed a significant reduction (P < 0.05). A high amount of splenocytes secreting IFNγ and IL-17 was seen in immunized mice with rCD44v (P < 0.05). Furthermore, a smaller size of lung metastases compared to the control mice groups was detected. CONCLUSION: The encapsulated rCD44v within the chitosan nanoparticles induced a significant immune response in mice and can establish significant protection against breast cancer. Therefore, it can be considered a vaccine candidate for breast cancer therapeutic modalities.

Olive oil-based quercetin nanoemulsion (QuNE)'s interactions with human serum proteins (HSA and HTF) and its anticancer activity.

The current study produced Quercetin nanoemulsions (QuNEs) for the purpose of improving Quercetin solubility in an aqueous polar condition and to analyze QuNE-protein formation (QuNE-human serum albumin (HSA) and QuNE-holo-transferrin (HTF)).QuNE was produced by utilizing an ultrasound-based emulsification method and was characterized by DLS, TEM, and SEM. Its interaction with HSA and HTF proteins was studied by analyzing the results of FRET and RLS spectroscopy, Stern-Volmer plotting, the Van't Hoff equation, CD spectroscopy, and molecular docking methods. Finally, QuNE's cytotoxic impact, cell death type induction, and antioxidant properties were evaluated by applying an MTT assay on a human hepatocyte cancer cell (HepG2), measuring Cas-3 gene expression, and conducting a DPPH antioxidant test, respectively. Compared to the non-entrapped Quercetin, Quercetin-entrapped nano-emulsions formed stable complexes with HSA and HTF by improving hydrophilic-hydrophobic interactions. The binding constant (BC), ΔH0, and ΔS0 indices for both the QuNE-HSA and QuNE-HTF complexes were measured at (4.92 × 105 and 11.99 × 104 M-1), (170.96 and -131.19 KJ.mol-1), and (-464.86 and 342.83J.mol-1K-1), respectively.QuNE lowered the HepG2 viability by up-regulating Cas-3 gene expression and thus inducing apoptosis. Moreover, a notable antioxidant impact on the QuNE was detected. Due to its ability in delivering Quercetin to HSA and HTF proteins and stabilizing their protein complexes, QuNE can be used as a suitable primary transporting agent whose formation of stable bio-accessible QuNE-HSA and -HTF protein complexes creates a safe and natural secondary delivery system, which has potential to be used as an efficient anticancer compound.Communicated by Ramaswamy H. Sarma.

New insights on the binding of butyl-paraben to trypsin: experimental and computational approaches.

Butyl-paraben (BP) is one of the most widely used preservatives in numerous foodstuffs, skin care products, and a variety of drugs, and trypsin is the main digestive enzyme, the research on the binding between the two is essential for human health. In the present paper, the effect of BP on trypsin has been explored using experimental and computational techniques to evaluate BP toxicity at the protein level. The obtained results from molecular docking and kinetic assay revealed BP was embedded in the hydrophobic cavity-S1 binding pocket of the enzyme to inhibit its activity by a competitive model. Intrinsic fluorescence of trypsin after interaction with BP revealed the static mode of quenching. FRET indicated that the distance of the enzyme to BP is 1.89 nm with high energy efficiency. Thermodynamic results proved that BP spontaneously bound to trypsin in an enthalpy-driven manner, the van der Waals interactions and H-bonds serving as the predominant forces in binding processes. CD spectroscopy and molecular dynamics (MD) simulation revealed that the trypsin structure transformed from the ß-Sheet structure to the unordered Coil structure upon interacting with BP. Resonance light scattering (RLS), synchronous fluorescence, and three-dimensional (3 D) spectroscopies further supported the alteration in the conformation of trypsin. Differential scanning calorimetry (DSC) showed that trypsin was somewhat destabilized in the presence of BP. Accordingly, all of the experimental data were confirmed by MD simulation.Communicated by Ramaswamy H. Sarma.

In silico designing and expression of novel recombinant construct containing the variable part of CD44 extracellular domain for prediagnostic breast cancer.

BACKGROUND: CD44, as a tumor-associated marker, can be used to detect stem cells in breast cancer. While CD44 is expressed in normal epithelial cells, carcinoma cells overexpress CD44. AIMS: In the current study, we designed a recombinant protein that included the variable component of the CD44 (CD44v) extracellular domain to apply in clinical diagnosis of breast cancer. METHODS: A total of 100 CD44v amino-acid residues were determined, and the structure was examined using bioinformatics tools. The construct was inserted into the PET28a vector and transformed in E. coli BL21(DE3). A nearly 12 kDa fusion protein was obtained by Ni-NTA affinity metal chromatography. Recombinant CD44v was examined by Western blotting, ELISA, and immunohistochemistry (IHC) assays. RESULTS: The findings revealed that the structure of rCD44v was stable, and its antigenic domain was exposed. The recombinant CD44v was confirmed by western blotting, and the presence of antibodies against recombinant CD44v protein in the patient's serum was detected by the ELISA. Our data demonstrated a link between CD44v serum levels and the prevalence of breast cancer. CONCLUSION: Assessments of antiCD44v antibodies with rCD44v could be a useful tool for identifying breast cancer in its early stages, which can lead to better outcomes.

Association of hypoxia-inducible factor 1 expressions with prognosis role as a survival prognostic biomarker in the patients with osteosarcoma: a meta-analysis.

BACKGROUND: Osteosarcoma, the most prevalent primary bone cancer, tends to relapse or metastasize quickly. Hypoxia-inducible factor-1 alpha (HIF-1α) affects tumor metabolism, differentiation, angiogenesis, proliferation, and metastasis. Many studies have investigated the possible inconsistent prognostic value of HIF-1 α. This study evaluated the correlation between HIF-1 α expression and prognosis in osteosarcoma patients. METHODS: : A total of 978 patients from 12 studies were followed up. A meta-analysis was conducted on articles investigating HIF-1 α prognostic value in osteosarcoma patients. The authors excluded articles with overlapping data, duplicate data, reviews, case reports, and letters that did not provide original data. Calculation of the hazard ratios (HR) and pooled risk ratios (RR) with corresponding 95% confidence intervals were used to determine the association degree (CIs). RESULTS: It was determined that HIF-1 α in osteosarcoma patients had a prognostic value based on the RRs and HRs. The results showed that high HIF-1 α expression was associated with a worse prognosis when compared to low or undetectable HIF-1 α expression. CONCLUSION: HIF-1 α overexpression was found to predict poor outcomes in osteosarcomas. The present study suggests that HIF-1α is a useful prognostic biomarker to predict OS in patients with osteosarcoma.

Study of FA12 peptide-modified PEGylated liposomal doxorubicin (PLD) as an effective ligand to target Muc1 in mice bearing C26 colon carcinoma: in silico, in vitro, and in vivo study.

OBJECTIVES: This study tried to achieve active targeting of Muc1 in cancer; the surface of PEGylated liposomal doxorubicin (PLD/Doxil®) was decorated with FA12 peptide. METHODS: According to docking results, FA12 was selected for this study, among four different peptides. MD simulation was also conducted as an additional confirmation of the binding interaction between FA12 and Muc1. Liposomal formulations were prepared; 1HNMR and HPLC techniques were used to verify peptide conjugation to DSPE-PEG2000-COOH. Afterward, DSPE-PEG2000-FA12 was post-inserted into the PLD at 50, 100, 200, and 400 peptides per liposome. The size, zeta potential, release profile, cytotoxicity (IC50), and cell uptake (using fluorescence microscopy and flow cytometry) were evaluated. In vivo biodistribution and antitumor activities were studied on mice bearing C-26 colon carcinoma. RESULTS: Cell uptake and cytotoxicity results revealed that PLD-100 (targeted PLD with 100 FA12 per liposome) could significantly enhance cellular binding. Furthermore, PLD-100 demonstrated higher antitumor efficacy, indicating more remarkable survival compared to PLD and other targeted PLDs. PLD-100 exhibited higher doxorubicin tumor accumulation compared to PLD. CONCLUSIONS: FA12 peptide is a promising targeting ligand for PLD to treat cancers with a high level of Muc1 expression and merits further investigations.

In this work, we used an antimicrobial peptide, FA12, to target Muc1 glycoprotein on the surface of colon cancer cells. The interaction between the peptide and Muc1 as a receptor was verified by molecular dynamics simulation. FA12 peptide was affixed to the surface of the liposomal form of doxorubicin (PLD) specifically to facilitate drug delivery transfer to the tumor site. The main benefits of this novel formulation were improvement of therapeutic efficacy and enhancement of survival time in mice bearing colon cancer.[Figure: see text].

The expression of long non-coding RNA LINC01389, LINC00365, RP11-138J23.1, and RP11-354K4.2 in gastric cancer and their impacts on EMT.

BACKGROUND: Epithelial cancers acquire the epithelial to mesenchymal transition (EMT), which leads tumor cells to invade and metastasize to adjacent and distant tissues. The mechanisms involved in EMT phenotype are controlled by numerous markers as well as signalling pathways. Recently, long non-coding RNAs (lncRNAs) were introduced that play the regulatory role in EMT via crosstalk with EMT-related transcription factors and signalling pathways. The present study aimed to investigate the expression of four lncRNAs in human GC and elucidate their probable role in EMT procedure and the pathogenesis of gastric cancer (GC). METHODS: The expression profile of lncRNAs (LINC01389, LINC00365, RP11-138J23.1, and RP11-354K4.2) and mRNAs (TWIST1, MMP13, MAML1, CD44s, and SALL4) between eighty-three GC and adjacent non-cancerous tissues were assessed by quantitative real-time PCR. RESULTS: The significant downregulation of LINC00365 (66.3%) and RP11-354K4.2 (62.7%) were observed in GC samples; while the upregulation of LINC01389, RP11-138J23.1, TWIST1, MMP13, MAML1, CD44s, and SALL4 were found in 67.5%, 45.8%, 56.6%, 44.6%, 59%, 55.4%, and 62.7% tumors samples at the mRNA level, respectively. Dysregulation of these lncRNAs and EMT-related markers was significantly related to each other in a variety of clinicopathological features of patients (P < 0.05), indicating positive correlations between LINC01389, LINC00365, RP11-138J23.1, and RP11-354K4.2 with EMT status in GC. CONCLUSION: These EMT-regulating lncRNAs may play a key role in transforming gastric epithelial to mesenchymal phenotype and can be novel therapeutic targets for GC. Our results highlight the importance of discovering new lncRNAs involved in gastric carcinogenesis. Detailed molecular mechanisms of these noncoding-coding markers in GC are urgently required.

Recent advances in the bio-application of microalgae-derived biochemical metabolites and development trends of photobioreactor-based culture systems.

Microalgae are microscopic algae in sizes ranging from a few micrometers to several hundred micrometers. On average, half of the oxygen in the atmosphere is produced by the photosynthetic process of microalgae, so the role of these microorganisms in the life cycle of the planet is very significant. Pharmaceutical products derived from microalgae and commercial developments of a variety of supplements extracted from them originate from a variety of their specific secondary metabolites. Many of these microalgae are a reservoir of unique biological compounds including carotenoids, antioxidants, fatty acids, polysaccharides, enzymes, polymers, peptides, pigments, toxins and sterols with antimicrobial, antiviral, antifungal, antiparasitic, anticoagulant, and anticancer properties. The present work begins with an introduction of the importance of microalgae in renewable fuels and biodiesel production, the development of healthy food industry, and the creation of optimal conditions for efficient biomass yield. This paper provides the latest research related to microalgae-derived substances in the field of improving drug delivery, immunomodulatory, and anticancer attributes. Also, the latest advances in algal biocompounds to combat the COVID-19 pandemic are presented. In the subject of cultivation and growth of microalgae, the characteristics of different types of photobioreactors, especially their latest forms, are fully discussed along with their advantages and obstacles. Finally, the potential of microalgae biomass in biotechnological applications, biofuel production, as well as various biomass harvesting methods are described.

The effect of angiotensin receptor blockers and angiotensin-converting enzyme inhibitors on progression of gastric cancer: systematic review and meta-analysis.

Angiotensin receptor blockers (ARB), as well as angiotensin-converting enzyme inhibitors (ACEI), are mostly used as therapy for hypertension and cardiovascular disease. However, they can increase the risk of cancer progression including gastric cancer. Here we aimed to analyze the assessment between ARB and ACEI on the progression of gastric cancer. Cochrane Library, PubMed and EMBASE were searched for articles and abstracts describing ARBs, ACEIs, and incidence of gastric cancer. Risk ratio, hazard ratio and 95% confidence interval (CI) were extracted from each outcome by using a random-effects model. Six studies met our inclusion criteria. These results demonstrated that there is a significant association between ARB with gastric cancer progression (risk ratio = 0.63; 95% CI, 0.5-0.7; P = 0.00; I 2 = 27.299; df (Q) = 2; Q-value = 2.75). However, there was not any link between ACEIs and gastric cancer development (risk ratio = 1.1; 95% CI, 0.92-1.31; P = 0.26; I 2 = 0.00; df (Q) = 3; Q-value = 1.26). All these findings indicated that using the ARBs has raised the progression of gastric cancer in these patients.

Inhibitory role of LINC00332 in gastric cancer progression through regulating cell EMT and stemness.

OBJECTIVE: Gastric cancer (GC) is one of the most common lethal malignancies worldwide. The molecular mechanisms underlying GC early detection are poorly understood. Identifying potential coding and non-coding markers and related pathways in the GC progression is essential. Some Long non-coding RNAs (lncRNAs) reportedly play vital roles during gastric GC development. However, the clinical significance and biological function of LINC00332 in GC remain largely unclear. METHODS: The gene expression patterns of GC from an RNAseq dataset (GSE122401) were retrieved from the Gene Expression Omnibus (GEO) database to recognize differentially expressed genes (DEGs) and lncRNAs (DELs) between normal and GC samples through several bioinformatic analysis. The expression of LINC00332 and MMP-13 as a target gene was quantified in fresh frozen tissues obtained from GC patients. In addition, we investigated the potential function of LINC00332 in silico and in vitro. RESULTS: The expressions of LINC00332 and MMP-13 were significantly downregulated and upregulated in GC tissues, respectively. A significant inverse correlation between LINC00332 and MMP-13 mRNA expression was observed in tumor samples. The mRNA expression level of mesenchymal markers, stem cell factors, and MMP genes were significantly decreased after the LINC00332 ectopic expression, while epithelial markers expression was significantly increased. The LINC00332 overexpression markedly repressed proliferation, migration, and invasion and did not induce apoptosis in AGS cells. In addition, LINC00332 overexpression notably promoted the E-cadherin protein expression. Moreover, LINC00332 significantly decreased the cisplatin resistance. CONCLUSION: Our findings indicated that LINC00332 may be a critical anti-EMT factor and provided a new efficient therapeutic strategy for GC treatment.

Fabrication, characterization and in vitro cell exposure study of zein-chitosan nanoparticles for co-delivery of curcumin and berberine.

For the first time, we synthesized the co-delivery nanopolymers using zein protein as the core and chitosan polysaccharide as the shell to deliver curcumin (Cur) and berberine (Ber) in MDA-MB-231 breast cancer cells. It has been shown that Cur and Ber altogether have synergistic effects on multiple cancers. Herein, the curcumin-zein-berberine-chitosan (Cur-Z-Ber-Ch) nanoparticles were fabricated and their organization procedure was reported. Physicochemical properties of synthesized nanoparticles were determined by Fourier transform infrared (FTIR) spectroscopy, XRD and fluorescence spectroscopy analyses. The nanoparticles included relatively small particles (d = 168.24 nm) with +36.76 mV zeta potential. The resulting nanoparticles had high entrapment efficiency (about 75%) for Cur and 60% for Ber. The Cur-Z-Ber-Ch nanoparticles represented ideal redispersibility and storage stability after 4 months. Drug release of the freeze-dried nanoparticles had pH-sensitive characteristic. In vitro cytoxicity assay demonstrated that Cur-Z-Ber-Ch nanoparticles induced elevated cytotoxic effect in MDA-MB-231 and A549 cancer cells. In vitro studies in MDA-MB-231 cells demonstrated that the Cur-Z-Ber-Ch nanoparticles could successfully increase cellular uptake and apoptosis with significant inhibition of IL-8 pro-inflammatory cytokines in comparison to the free Cur + Ber bioactive molecules. These bio-nanoparticles are the co-delivery vehicle for Cur and Ber which could be beneficial for participating them into pharmaceutical products.

A novel approach towards obesity: The use of a bacterial product, gassericin A, in 3T3-L1 cells.

BACKGROUND AND OBJECTIVE: The problem of obesity and its related complications are adversely affecting human society. We studied the effects of gassericin A, a bacteriocin produced by the intestinal bacteria, on adipocyte differentiation and development. DESIGN: Gassericin A was purified from Lactobacillus gasseri LA39 and was added to the culture medium of 3T3-L1 cells in two phases: Phase 1, 3T3-L1 cells were incubated with gassericin A while being induced to adipocytes (days 1-7); phase 2, the cells were incubated with the bacteriocin after being induced to adipocytes (days 8-12). The resultant changes in the pattern of expression of some of the important genes involved in adipogenesis were evaluated by RT-qPCR. The viability of cells and their numbers were also studied. RESULTS: In phase 1 of the study, the levels of transcripts for stearoyl CoA desaturase (SCD-1), zinc finger protein 423 (zfp-423), and glucose transporter 4 (GLUT4) genes were significantly reduced, while that of 422ap2 gene showed a significant increment (p < 0.05). In phase 2, the zfp-423 gene showed a reduction of expression and the 422ap2 gene showed an increase in expression (p < 0.05). The other genes including UCP-1 and TNF-α did not show any significant changes in neither of the groups. Gassericin A did not affect the morphology or viability of the cells, however, the numbers of cells had nearly doubled in the treatment groups. CONCLUSION: It seems that gassericin A could significantly alter the properties of adipocytes while they are in the process of development and after they have developed.

Evaluation of the in vivo antihypertensive effect and antioxidant activity of HL-7 and HL-10 peptide in mice.

BACKGROUND: The tendency to use bioactive peptides has increased in recent decades, and research would be essential for recognizing the therapeutic effects of peptides present in animals or food resource. In this study, the in vivo antioxidant and antihypertensive properties of peptides HL-7 with the sequence of YLYELR and HL-10 with the sequence of AFPYYGHHLG were identified from scorpion venom of H. lepturus were evaluated. METHODS AND RESULTS: To study the in vivo effects of peptides, D-galactose-induced and DOCA salt-induced mice models were used. The results of the antioxidant assay for both peptides showed that the activity of serum and liver catalase (CAT), as well as superoxide dismutase (SOD) enzymes, was significantly decreased in the D-galactose-induced group (NC), while MDA levels were increased in serum and the liver tissue samples (p < 0.01). Compared with the D-galactose-induced mice, the peptide treated mice group had a higher activity of antioxidant enzymes namely CAT and SOD, as well as a lower lipid peroxidation level. Also, the results of antihypertensive activity for both peptides showed that systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the mice treated with the HL-7 and HL-10 peptides were significantly reduced in a dose-dependent manner (p < 0.01). The administration of the HL-7 peptide at doses of 2 mg/kg BW (LP1), 5 mg/kg BW (-IP1) and 15 mg/kg BW (HP1) significantly diminished the mean arterial blood pressure (MAP) by 11 mmHg, 31 mmHg and 40.47 mmHg, respectively. Accordingly, treatment of mice with the HL-10 peptide at doses of 2 mg/kg BW (LP2), 5 mg/kg BW (IP2) and 15 mg/kg BW (HP2) considerably lowered the MAP by 8 mmHg, 18.3 mmHg and 21.93 mmHg, respectively. CONCLUSION: Our findings suggest that both the HL-7 and HL-10 peptides could be potentially utilized as antihypertensive and antioxidant components.