The prevalence of 17 common respiratory viruses in patients with respiratory illness but negative for COVID-19: A cross-sectional study.

Background and Aims: Second to COVID-19 pandemic, other viral respiratory infections are still important causes of human diseases or co-infections. Hence, the present study was carried out to investigate the common respiratory viruses in patients with respiratory illness diagnosed negative for severe acute respiratory syndrome coronavirus-2 in primary screening. Methods: In a cross-sectional study, a real-time PCR was carried out using HiTeq. 17 Viro Respiratory pathogen One Step RT-PCR Kit (Genova, Bonda Faravar, Bioluence, Tehran, Iran). Results: A total of 311 individuals (mean age ± SD: 48.2 ± 21.7 years, range: 1-97 years) underwent second PCR. Among these, 161 (51.7%) were female. In total, 55 (17.6%) cases (mean age ± SD: 45.7 ± 18.1 years) were found positive for respiratory viruses panel in the second PCR. The HCoV-OC43/HKU1 was in 5.4% (17/311), Flu A in 4.5% (14/311), HCoV-229E/NL63 in 2.8% (9/311), HMPV in 1.9% (6/311), HPiV 1, 2, 3 in 1.2% (4/311), HRSV in 0.9% (3/311), and HAdV in 0.6% (2/311) of the cases studies. Also, co-infection was detected in 4 samples (1.2%). In addition, sore throat (0.028), headache (p = 0.016), and body pain (p = 0.0001) were statistically the most significant symptoms in studied cases. Conclusion: According to the findings of our study, respiratory virus infections and co-infections were 17.6% and 1.2% frequent, respectively. Interestingly, nearly half of our positive cases (47.2%) were identified by coronaviruses (ОС43, Е229, NL63, and HKUI), followed by influenza A virus (25.4%). However, for more comprehensive results, we recommend using greater sample size.

Simvastatin-Loaded Nanoniosome Protects H9c2 Cells from Oxygen-Glucose Deprivation/ Reperfusion Injury by Downregulating Inflammation.

Background: Simvastatin (SIM) has anti-inflammatory and antioxidant properties against cardiac ischemia/reperfusion injury (I/RI). However, it suffers from low bioavailability and a short half-life. Nanoniosomes are novel drug delivery systems that may increase SIM effectiveness. The present research evaluates the impact of SIM-loaded nanoniosomes on the oxygen-glucose deprivation/reperfusion (OGD/R) injury model of H9c2 cells. Methods: Cells were seeded based on five groups: (1) control; (2) OGD/R; (3) OGD/R receiving SIM; (4) OGD/R receiving nanoniosomes; and (5) OGD/R receiving SIM loaded nanoniosomes. OGD/R injury of the H9c2 cells was treated with SIM or SIM loaded nanoniosomes. Cell viability, two inflammatory factors, necroptosis factors, along with HMGB1 and Nrf2 gene expressions were assessed. Results: The cells treated with SIM loaded nanoniosomes showed a significant elevation in the cell viability and a reduction in HMGB1, Nrf2, TNF-α, IL-1ß, RIPK1, and ROCK1 expression levels compared to the OGD/R and SIM groups. Conclusion: Based on our findings, nanoniosomes could safely serve as a drug delivery system to counterbalance the disadvantages of SIM, resulting in improved aqueous solubility and stability.

Tyrosol-loaded Nano-niosomes Attenuate Diabetic Injury by Targeting Glucose Metabolism, Inflammation, and Glucose Transfer.

INTRODUCTION: Regarding the increasing prevalence of type 2 diabetes, it has become a global concern, making it imperative to control. Chemical drugs commonly recommended for diabetes treatment cause many complications and drug resistance over time. METHODS: The polyphenol tyrosol has many health benefits, including anti-diabetic properties. Tyrosol's efficacy can be significantly increased when it is used as a niosome in the treatment of diabetes. In this study, Tyrosol and nano-Tyrosol are examined for their effects on genes implicated in type 2 diabetes in streptozotocin-treated rats. Niosome nanoparticles containing 300 mg surfactant (span60: tween60) and 10 mg cholesterol were hydrated in thin films with equal molar ratios. After 72 hours, nano-niosomal formulas were assessed for their physicochemical properties. MTT assays were conducted on HFF cells to assess the cellular toxicity of the nano niosome contacting optimal Tyrosol. Finally, the expression of PEPCK, GCK, TNF-ɑ, IL6, GLUT2 and GLUT9 was measured by real time PCR. RESULTS: Physiochemical properties of the SEM images of niosomes loaded with Tyrosol revealed that the nanoparticles had a vehicular structure. In this study, there were two stages of release: initial release (8 hours) and sustainable release (72 hours). Meanwhile, free form drugs were considerably more toxic than niosomal drugs in terms of their cellular toxicity. An in vivo comparison of oral Tyrosol gavage with nano-Tyrosol showed a significant increase in GCK (P<0.001), GLUT2 (P<0.001), and GLUT9 (P<0.001). Furthermore, nano-Tyrosol decreased the expression of TNF-ɑ (P<0.05), PEPCK (P<0.001), and IL-6 (P<0.05) that had been increased by diabetes mellitus. The results confirmed nano-Tyrosol's anti-diabetic and anti-inflammatory effects. CONCLUSION: These findings suggest that nano-Tyrosol has potential applications in diabetes treatment and associated inflammation. Further research is needed to better understand the mechanism of action.

Nano-Stevia reduces the liver injury caused by streptozotocin (STZ)-induced diabetes in rats by targeting PEPCK/GCK genes, INSR pathway and apoptosis.

Objectives: Extensive application of stevia in the treatment of type 2 diabetes mellitus (DM) has been proven by a large number of previous studies. We prepared stevia loaded in nanoniosomes (nanostevia) to improve its bioavailability, functionality, and stability and explore its protective effects and underlying mechanisms in the liver of STZ-induced diabetic rats. Methods: Single-dose intraperitoneal injection of STZ (50 mg/kg body weight) was used to establish diabetic model. The mRNA levels of PEPCK and GCK genes and the protein level of INSR were evaluated by Real time-PCR and Western blot assays, respectively. TUNEL assay was used to detect apoptotic cell death in the liver tissue. Results: Diabetic rats exhibited significantly reduced levels of INSR (*** P < 0.001) as well as elevated levels of PEPCK (*** P < 0.001). Both stevia and nano-stevia were capable of increasing levels of GCK and INSR and reducing levels of PEPCK (## P < 0.01 and ### P < 0.001, respectively). In addition, significantly increased number of apoptotic cell death was seen in the liver tissue of diabetic rats (*** P < 0.001) which was markedly mitigated by treatment with both Stevia and nano-Stevia (#P < 0.05 and ## P < 0.01, respectively). Conclusion: Both stevia and nano-stevia demonstrates potent anti-apoptotic activity in the liver tissue of diabetic rats by targeting PEPCK/GCK genes and INSR pathway. These finding show that nano-stevia has more potential to reduce the liver injury caused by STZ-induced diabetes in rats and hence can be considered a valid agent and alternative therapy for attenuating complications of type 2 DM. Supplementary Information: The online version contains supplementary material available at 10.1007/s40200-023-01278-2.

Efficient targeting of HIF-1α mediated by YC-1 and PX-12 encapsulated niosomes: potential application in colon cancer therapy.

A number of molecular biofactors have been documented in pathogenesis and poor prognosis of colorectal cancer (CRC). Among them, the Hypoxia-Inducible Factor (HIF-1a) is frequently reported to become over-expressed, and its targeting could restrict and control a variety of essential hallmarks of CRC. Niosomes are innovative drug delivery vehicles with the encapsulating capacity for co-loading both hydrophilic and hydrophobic drugs at the same time. Also, they can enhance the local accumulation while minimizing the dose and side effects of drugs. YC-1 and PX-12 are two inhibitors of HIF-1a. The purpose of this work was to synthesize dual-loaded YC-1 and PX-12 niosomes to efficiently target HIF-1α in CRC, HT-29 cells. The niosomes were prepared by the thin-film hydration method, then the niosomal formulation of YC-1 and PX-12 (NIO/PX-YC) was developed and optimized by the central composition method (CCD) using the Box-Behnken design in terms of size, polydispersity index (PDI), entrapment efficiency (EE). Also, they are characterized by DLS, FESEM, and TEM microscopy, as well as FTIR spectroscopy. Additionally, entrapment efficiency, in vitro drug release kinetics, and stability were assessed. Cytotoxicity, apoptosis, and cell cycle studies were performed after the treatment of HT-29 cells with NIO/PX-YC. The expression of HIF-1αat both mRNA and protein levels were studied after NIO/PX-YC treatment. The prepared NIO/PX-YC showed a mean particle size of 185 nm with a zeta potential of about-7.10 mv and a spherical morphology. Also, PX-12 and YC-1 represented the entrapment efficiency of about %78 and %91, respectively, with a sustainable and controllable release. The greater effect of NIO/PX-YC than the free state of PX-YC on the cell survival rate, cell apoptosis, and HIF-1α gene/protein expression were detected (p < 0.05). In conclusion, dual loading of niosomes with YC-1 and PX-12 enhanced the effect of drugs on HIF-1α inhibition, thus boosting their anticancer effects.

Oral administration of nano-tyrosol reversed the diabetes-induced liver damage in streptozotocin-induced diabetic rats.

Objectives: The present study was designed to evaluate the effects of Tyrosol and Nano-tyrosol on the cellular arrangement, collagen disposition, protein level of insulin receptor (INSR), and superoxide dismutase (SOD) activity in both control and streptozotocin-induced diabetic rats. Methods: Type 2 Diabetes (T2D) was induced in rats by a single intraperitoneal injection of streptozotocin (50 mg/kg). Experimental rats were administered Tyrosol and Nano-tyrosol 1 ml intra-gastrically at a dose of 20 mg/kg once a day for 30 days. Then, rats were sacrificed according to ethical principles. Livers were removed and processed for histological studies using the paraffin technique. Furthermore, non-paraffin sections were used for the INSR-1 western blot technique. Results: At the end of the experiments, the rats in diabetic control and plain niosome groups exhibited a significant increase in collagen disposition (p < 0.001), and apoptotic cells (p < 0.001), as well as decreased total protein levels of INSR (p < 0.001), and SOD activity (p < 0.001) in the hepatic cells. Oral administration of Tyrosol and Nano-tyrosol to diabetic rats reversed all the above-mentioned parameters to near normal levels (p < 0.001). Nano-tyrosol showed the highest significant effect rather than Tyrosol. Conclusion: The results of the present study suggested the beneficial effects of Tyrosol and especially Nano-tyrosol on decreasing the adverse effects of diabetes.

DNAzyme loaded nano-niosomes attenuate myocardial ischemia/reperfusion injury by targeting apoptosis, inflammation in a NF-κB dependent mechanism.

Although DNAzymes have been found to reduce injury after myocardial ischemia/reperfusion (MI/R), their efficiency have been limited due to rapid degradation in vivo. Thus, this study was conducted to extend their half-life by encapsulation into nano­niosomes and examine their cardioprotective effects in a rat model of myocardial infarction (MI). In order to synthesize nano­niosomes, surface active agent film hydration method was used. Characterization of nano­niosomes was performed using the atomic force microscopy (AFM). In order to establish MI/R model in rats, left anterior descending coronary artery (LAD) was ligated for 30 min. A single dose (150µL) of drug formulations was injected into the infarcted region. The cardiac function was evaluated using echocardiography. The expression of pro-inflammatory cytokines, apoptotic factors, and nuclear factor-κB (NF-κB) were evaluated using Western blot and immunohistochemistry, respectively. Particle size of only nano-niosomes was in the range of 60-90 nm, while a shift to 70-110 nm was seen after DNAzyme encapsulation. MI rats treated with DNAzyme­loaded nano­niosomes could markedly reduce Bax, caspase3, TNF-α, IL-1ß, and NF-κB as well as increase Bcl-2 compared to only MI/R group. Collectively, our finding show that nano­niosomes can be considered excellent drug delivery platforms to extend half-life and stability of DNAzyme, when it is used to reduce myocardial I/R injury.

Intra-gastrically administration of Stevia and particularly Nano-Stevia reversed the hyperglycemia, anxiety, and memory impairment in streptozotocin-induced diabetic rats.

Type II diabetes mellitus is a group of metabolic disorders considered chronic hyperglycemia resulting from deficits in insulin secretion or insulin function. This disease usually links with various psychological problems such as anxiety and cognitive dysfunctions. Stevia (Stevia rebaudiana Bertoni) is a natural and healthy substitute sweetener for sugar and artificial sweeteners. It has become essential for human diets and food manufacturers. The aim of this research was to investigate the effects of Stevia and Nano-stevia on the regulation of anxiety and memory processes in male diabetic rats. The elevated plus-maze (EPM) test-retest procedure was used to assess anxiety and memory in male diabetic rats. The findings exhibited that induction of diabetes caused a distorted cellular arrangement in the liver tissue of male rats. On the other hand, intra-gastrically administration of Stevia (1 ml/kg) and nano-Stevia (1 ml/kg) indicated a normal appearance in the liver tissue of male diabetic rats. Moreover, induction of diabetes caused the augmentation of blood glucose, reduction in time spent in%open-arm time (%OAT) on the test day, and enhancement of%OAT on the retest day. Therefore, induction of diabetes in rats produced hyperglycemia, anxiogenic effect, and memory impairment and these responses were reversed by drug treatment. Furthermore, intra-gastrically application of Stevia (1 ml/kg) and nano-Stevia (1 ml/kg) reversed the hyperglycemia, anxiogenic effect, and memory impairment in male diabetic rats. Interestingly, Nano-Stevia exhibited the highest significant response rather than Stevia. In conclusion, the results of this research suggested the beneficial properties of Stevia and particularly Nano-Stevia on inducing anti-diabetic effects, anxiolytic behavior, as well as memory improvement in male diabetic rats.

Chitosan -based nanoniosome for potential wound healing applications: Synergy of controlled drug release and antibacterial activity.

This study aims to develop a niosomal platform which can delivery drugs such as tetracycline hydrochloride (TCH) to treat bacterial infections in wounds. To this end, chitosan (CS) was used to obtain a controlled drug release and at the same time antibacterial activity. By design of experiments the niosome encapsulated TCH (TCH-Nio) were optimized for their particle size and encapsulation efficiency, followed by analysis of the release profile of TCH and stability of TCH-Nio and TCH-Nio@CS. The antibacterial activity and cytotoxicity of the fabricated nanoparticles were investigated as well. The release rate of TCH from TCH-Nio@CS in all conditions is less than TCH-Nio. In addition, higher temperature increases the release rate of drug from these formulations. The size, polydispersity index, and encapsulation efficacy of TCH-Nio and TCH-Nio@CS were more stable in 4 °C compared to 25 °C. TCH, TCH-Nio, and TCH-Nio@CS had MIC values of 7.82, 3.91, and 1.95 µg/mL for Escherichia coli, 3.91, 1.95, and 0.98 µg/mL for Pseudomonas aeruginosa, and 1.96, 0.98, and 0.49 µg/mL for Staphylococcus aureus, respectively. Coating of chitosan on niosome encapsulated TCH (TCH-Nio@CS) led to a reduced burst release of TCH from niosome (TCH-Nio), and enabled 2-fold higher antibacterial and anti-biofilm activity against the tested bacterial pathogens E. coli, P. aeruginosa and S. aureus, compared to the uncoated TCH-Nio, and 4-folder higher than the TCH solution, suggesting the synergetic effect of niosome encapsulation and chitosan coating. Moreover, the formulated niosomes displayed no in vitro toxicity toward the human foreskin fibroblast cells (HFF). Both TCH-Nio and TCH-Nio@CS were found to down-regulate the expression of certain biofilm genes, i.e., csgA, ndvB, and icaA in the tested bacteria, which might partially explain the improved antibacterial activity compared to TCH. The obtained results demonstrated that TCH-Nio@CS is capable of controlled drug release, leading to high antibacterial efficacy. The established platform of TCH-Nio@CS enlighten a clinic potential toward the treatment of bacterial infections in skin wounds, dental implants and urinary catheter.

Nesfatin-1 attenuates injury in a rat model of myocardial infarction by targeting autophagy, inflammation, and apoptosis.

Nesfatin-1 plays an important role in the modulation of heart performance. However, it remains unclear how nesfatin-1 contributes to cell survival in acute myocardial infarction (MI). A rat model of MI was established via ligation of left anterior descending coronary artery (LAD) for 30 min and 20 µg/kg concentration of nesfatin-1 was intraperitoneally infused prior to reperfusion. At 24 h after reperfusion, oxidative stress markers, the expression of caspase3, beclin-1, pro-inflammatory cytokines, and the mRNA levels of Bax and Bcl-2 were evaluated. Results showed that nesfatin-1 markedly restored GSH content and SOD activity as well as reduced MDA levels compared to only the MI group (p < .05). Likewise, nesfatin-1 contributed to cell survival by inhibiting autophagy and apoptosis markers such as caspase3 and Bax (p < .05). Collectively, these findings support the idea that nasfatin-1 can be used as a good candidate to treat MI by targeting oxidative stress, apoptosis, and autophagy.

In vitro and in silico characterization of a novel glutamate carboxypeptidase from Cohnella sp. A01.

Glutamate carboxypeptidase is a bacterial enzyme of metallopeptidase superfamily. This enzyme is an exo-peptidase that catalyzes the hydrolysis of glutamate residues at the C-terminus of folic acid. The rCP302 is a novel zinc ion-dependent recombinant glutamate carboxypeptidase derived from a thermophilic bacterium, Cohnella sp. A01 (PTCC No: 1921). By simulating the structure of rCP302, analyzing its activity in various environmental settings, and contrasting it with that of related enzymes, we wanted to evaluate the heterologous production, purification, and characterization of this enzyme. The bioinformatics study showed that rCP302 had maximum similarity to M20 family of metallopeptidases. The purified rCP302 molecular weight was about 41.6 kDa. The optimum temperature and pH for the catalytic activity of rCP302 were 50 °C and 7.2, respectively. Fluorescence spectroscopy data elucidated the secondary structure of rCP302 and determined conformational changes caused by alterations in ambient conditions. Using folate as a substrate, Km and specific activity values were calculated as 0.108 µM and 687 µmol/min/mg, respectively. The enzyme activity was strongly inhibited when EDTA sequestered zinc ions. The half-life of this enzyme at 30 °C was 2012 min. Regarding the ability of rCP302 to degrade folic acid, and its long half-life at 37 °C, the normal temperature of many mammals, this enzyme can be introduced for further study for use in the pharmaceutical industry.

Association of D299G Polymorphism of TLR4 Gene and CagA Virulence Factor of H. pylori among the Iranian Patients with Colorectal Cancer.

Background: Colorectal cancer (CRC) represents 9% of all malignancies globally. TLR4 gene defenses against Helicobacter pylori infection (HPI), so its mutations are a risk factor for CRC. As there is a correlation between (HPI) and gastric cancer, we investigated whether there is an association between CagA virulence factor in HPI and D299G polymorphism of TLR4 gene with developing CRC among Iranians. Methods: This retrospective study included 85 biopsies of confirmed colorectal lesions out of 230 subjects, which were divided into two age groups. Single nucleotide polymorphism (SNP) D299G in the TLR4 gene was assessed using Tetra-primer ARMS-PCR. The expression of TLR4 and the CagA virulence factor in H.pylori was assessed using real-time PCR (RT-PCR). Results: Chi-squared test showed genotype frequencies of GG were 79% and 62%in patients 51> and 51

Simvastatin-loaded nano-niosomes efficiently downregulates the MAPK-NF-κB pathway during the acute phase of myocardial ischemia-reperfusion injury.

BACKGROUND: Simvastatin can potentially mitigate acute inflammatory phase of myocardial ischemia-reperfusion injury. However, these effects negatively influenced by its poor bioavailability, low water solubility and high metabolism. Here, we investigated the effects of SIM-loaded nano-niosomes on a rat model of MI/R injury to find a drug delivery method to tackle the barriers. METHODS: Nano-niosomes' characteristics were identified using dynamic light scattering and transmission electron microscopy. Fifty male Wistar rats were divided into five groups: Sham; MI/R; MI/R + nano-niosome; MI/R + SIM; MI/R + SIM-loaded nano-niosomes. Left anterior descending artery was ligated for 45 min, and 3 mg/kg SIM, nano-niosomes, or SIM-loaded nano-niosomes was intramyocardially injected ten min before the onset of reperfusion. ELISA assay was used to assess cardiac injury markers (cTnI, CK-MB) and inflammatory cytokines (TNF-α, IL-6, TGF-ß, MPC-1). Expression level of MAPK-NF-κB and histopathological changes were evaluated by western blot and hematoxylin & eosin staining, respectively. RESULTS: the size of nano-niosome was 137 nm, reached to 163 nm when simvastatin was loaded. To achieve optimized niosomes span 80, a drug/cholesterol ratio of 0.4 and seven min of sonication time was applied. Optimized entrapment efficiency of SIM-loaded nano-niosomes was 98.21%. Inflammatory cytokines and the expression level of MAPK and NF-κB were reduced in rats receiving SIM-loaded nano-niosomes compared to MI/R + SIM and MI/R + SIM-loaded nano-niosomes. CONCLUSION: Our results showed that SIM-loaded nano-niosomes could act more efficiently than SIM in alleviating the acute inflammatory response of reperfusion injury via downregulating the activation of MAPK-NF-κB.

Human amniotic membrane mesenchymal stem cells exert cardioprotective effects against isoproterenol (ISO)-induced myocardial injury through suppression of inflammation and modulation of inflammatory MAPK/NF-κB pathway.

A common cause of mortality around the world is ischemic myocardial injury. The study was conducted to examine the ability of amniotic membrane mesenchymal stem cells (AMSCs) for protection against isoproterenol (ISO)-induced myocardial injury and attempted to show the possible mechanisms by which AMSCs that can be linked to inhibition of inflammation by targeting inflammatory MAPK/NF-κB pathway. Model was established by subcutaneous injection of 170 mg/kg/day of ISO for four consecutive days. Flow cytometry and echocardiography were carried out to evaluate characterization of hAMSCs and cardiac function, respectively. The expression of inflammatory cytokines was determined using ELISA assay. The activities of NF-κB and phosphorylated p38 MAPK were measured using immunohistochemical assessments. The results showed that ISO administration was resulted in cardiac dysfunction, increased levels of inflammatory cytokines that reversed by intramyocardially administration of AMSCs (P < 0. 05). Cardioprotective effects of AMSCs were associated with a significant decreased expression of NF-κB and reduced levels of phosphorylated p38 MAPK (P < 0. 05). In conclusion, our finding showed that intramyocardially administration of AMSCs could contribute to improvement of heart function and inhibition of inflammation in the site of injury by targeting inflammatory MAPK/NF-κB pathway.

Simvastatin-loaded nano-niosomes confer cardioprotection against myocardial ischemia/reperfusion injury.

Although simvastatin (SIM) has been proven to be a powerful agent against myocardial ischemia/reperfusion (MI/R) injury, poor water solubility, short half-life, and low bioavailability have made it futile while using conventional drug delivery system. Hence, this study aims to investigate therapeutic efficacy of SIM-loaded nano-niosomes on MI/R injury. Surface active agent film hydration method was used to synthesize nano-niosomes. The physicochemical properties of nano-niosomes were characterized using dynamic light scattering (DLS) and transmission electron microscopy (TEM). Moreover, niosomes were characterized in entrapment efficiency (EE) and releasing pattern. Male Wistar rats were assigned into five groups (sham, MI/R, SIM, nano-niosomes, and SIM-loaded nano-niosomes). To induce MI/R, left thoracotomy was performed along mid-axillary line. The LAD ligation lasted for 45 min. A single dose (3 mg/kg) of drug formulations was injected into myocardial. Echocardiography was performed to evaluate cardiac function. The expression of the necroptosis markers was evaluated using western blot assay. Particle size of only nano-niosomes was about 137 nm, whereas a shift to 163 nm was observed in nano-niosomes containing SIM. Optimized niosomes were achieved by span 80, drug to cholesterol ratio of 0.4 with 7-min sonication time. EE of optimized nano-niosomes containing SIM was 98.21%. The effects of nano-niosomes containing on improving cardiac function and inhibiting necroptosis pathway was more efficient than the SIM group. Our findings have suggested that nano-niosomes can be applied as a notable drug delivery method to augment stability, bioavailability, and therapeutic efficacy of SIM, when it used against myocardial I/R injury.

Which one of the thermal approaches (heating DNA or cells) enhances the gene expression in mammalian cells?

OBJECTIVES: Heat treatment as a physical method could increase the cellular uptake of nucleic acids. In this study, the effects of heat shock were evaluated to enhance the transfection efficiency of three plasmid DNAs into HeLa and TC-1 cancerous, and HEK-293 T and Vero non-cancerous cell lines using lipofectamine 2000 reagent. METHODS: Two methods of cell- and DNA-based heat treatment were used. Heating DNA solution was performed at 94 °C for 5, 10 and 15 min, and also 72 °C for 30, 60 and 120 min, individually. Moreover, heating the cells was done by incubation at 42 °C for 2 h in different times such as before, during and after DNA transfection. RESULTS: Our data showed that the conformation of plasmid DNAs was changed at different temperatures with increasing time. The heat-treated plasmid DNAs (94 °C for 10 min or 72 °C for 30 min) indicated higher transfection efficiency than untreated plasmid DNAs (p < 0.05). Furthermore, heat treatment of cells before and during the transfection was higher than untreated cells (p < 0.01). Our results demonstrated that DNA transfection efficiency in cancerous cells was less than non-cancerous cells (p < 0.01). CONCLUSION: Generally, these findings showed that transfection mediated by thermal stimulation could enhance gene transfection in mammalian cell lines.

Amniotic membrane mesenchymal stem cells labeled by iron oxide nanoparticles exert cardioprotective effects against isoproterenol (ISO)-induced myocardial damage by targeting inflammatory MAPK/NF-κB pathway.

The aim of the present study is to investigate the protective effects of human amniotic membrane-derived mesenchymal stem cells (hAMSCs) labeled by superparamagnetic iron oxide nanoparticles (SPIONs) against isoproterenol (ISO)-induced myocardial injury in the presence and absence of a magnetic field. ISO was injected subcutaneously for 4 consecutive days to induce myocardial injury in male Wistar rats. The hAMSCs were incubated with 100 µg/ml SPIONs and injected to rats in magnet-dependent and magnet-independent groups via the tail vein. The size and shape of nanoparticles were determined by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Prussian blue staining was used to determine cell uptake of nanoparticles. Myocardial fibrosis, heart function, characterization of hAMSCs, and histopathological changes were determined using Masson's trichrome, echocardiography, flow cytometry, and H&E staining, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to the expression pro-inflammatory cytokines. Immunohistochemistry assay was used to determine the expression of nuclear factor-κB (NF-κB) and the Ras/mitogen-activated protein kinase (MAPK). SPION-labeled MSCs in the presence of magnetic field significantly improved cardiac function and reduced fibrosis and tissue damage by suppressing inflammation in a NF-κB/MAPK-dependent mechanism (p < 0. 05). Collectively, our findings demonstrate that SPION-labeled MSCs in the presence of magnetic field can be a good treatment option to reduce inflammation following myocardial injury. Graphical abstract.

Anti-regulatory T cell vaccines in immunotherapy: focusing on FoxP3 as target.

Anti- tumor vaccination elicits imperfect immune responses against tumor cells; that is related to the presence of suppressive obstacles in the tumor microenvironment. The main members of suppressive milieu of tumor are heteroogenous groups of immune cells in which regulatory T cell is a substantial component. Tregs express different immunomodulatory molecules such as FoxP3. Transcription factor, FoxP3, is a specific intracellular marker of Treg and crucial for Treg development. Therefore it is an attractive target for cancer treatment. This article reviews some recent anti-Treg vaccine focusing on FoxP3 to ameliorate anti-tumor immune responses. Among them, fusion vaccine of FoxP3-Fc(IgG) recombinant DNA vaccine and its accordant protein vaccine represents effective results.