javad verdi

 The activities and functions of natural killer (NK) cells are regulated by a limited repertoire of activating and inhibitory receptors. Thus, we provided a study of inhibition of the NKG2A using monoclonal antibodies (mAbs), and as a primary endpoint, we evaluated whether it can be translated to enhance adoptive NK cell immunotherapy, as the secondary endpoint, we investigated safety and feasibility.

 In this study, we investigated the safety of anti-NKG2A-pretreated NK cells in improving ADCC function to manage hepatocellular carcinoma (HCC). After a conditioning regimen, we initiated a pilot study of expanded donor haploidentical NK cell infusion. Patients received a fludarabine/cyclophosphamide conditioning followed by adoptive immunotherapy with IL2–activated haploidentical NK cells. Anti-NKG2A pretreated NK cells were infused on days 0,+5, and+10 post-conditioning regimens at a dose of 7×108 cells (n=3). The median follow-up was 4 months for all patients.

 Although all patients were alive at the last follow-up, two of them showed progressive disease and an increase in tumor size. In addition, all patients showed a relative decrease in alpha-fetoprotein (AFP) expression levels after one month.

 This study demonstrated the safety and feasibility of infusing high doses of ex vivo expanded NK cells after conditioning with transient side effects.

Natural killer (NK) cells are crucial components of the innate immune system and have emerged as significant players in the pathogenesis of heart diseases. This review discusses recent findings regarding the multifaceted roles of NK cells in various cardiac conditions, including coronary artery disease, myocardial infarction, heart failure, myocarditis, and heart transplantation. It outlines the NK cell subsets, particularly CD56-bright and CD56-dim variations, their functional characteristics, cytokine profiles, and the inflammatory pathways they are involved. The review discusses both the beneficial and detrimental effects of NK cell activity on cardiac pathology by underlining their participation in immune regulation, tissue repair, and graft rejection dynamics. Additionally, we have addressed the impact of NK-cell–oriented environmental signals and discussed potential therapeutic approaches, such as immunomodulatory and anti-inflammatory strategies targeting NK cells. This review was therefore geared towards integrating available studies in understanding NK cell dynamics in heart disease and offering insights for future clinical interventions.

The important role of SMAD6 and several microRNAs (miRNAs), such as miR-17-5p, miR-26b-5p, and miR-32-5p, has been demonstrated in controlling the proliferation and differentiation of cardiomyocytes (CMs). Hence, this study was designed to assess the role of these regulatory factors in cardiac cell generation from human endometrium-derived mesenchymal stem cells (hEMSCs).

To induce transdifferentiation into CMs, hEMSCs were treated with a cardiac-inducing medium containing 5-azacytidine and bFGF for 30 days. Immunofluorescence staining and qRT-PCR, respectively, were used to measure the protein levels of SMAD6 and the mRNA expression of SMAD6 and the three miRNAs every six days.

Our findings demonstrated the mesenchymal stem cell properties of hEMSCs and their ability to differentiate into various types of mesenchymal stem cells. The differentiated hEMSCs exhibited morphological features resembling CMs. During the induction period, the number of positive cells for SMAD6 protein and the expression level of miR-26b-5p increased and peaking on days 24 and 30, while the expression levels of miR-17-5p and miR-32-5p decreased. The Pearson correlation coefficients revealed that SMAD6 level is inversely correlated with miR-17-5p and miR-32-5p and directly correlated with miR-26b-5p.

Our results indicate that miR-17-5p, miR-26b-5p, miR-32-5p, and SMAD6 are potentially involved in the molecular signaling pathways of transdifferentiation of hEMSCs to CMs.

Context: 

Adoptive T-cell therapy with chimeric antigen receptor (CAR) has shown tremendous progress in hematological cancers. However, some obstacles, such as high price tag, cytokine release syndrome, inability to penetrate solid tumors, and manufacturing complexity limit the wide application of this therapy. Natural killer (NK) cells can kill target cells via mechanisms similar to those of CD8+ cytotoxic T cells; therefore, CAR-NK cell therapy is a promising strategy for cancer treatment.

Evidence Acquisition:

 In this manuscript, all articles published in English regarding CAR-NKs and their application for the treatment of different types of cancers were collected from several databases, including PubMed, Scopus, and Google Scholar, using related keywords such as "Cancer, CAR construction, NK cells, and CAR-NK cells".

Compared with CAR-T cells, CAR-NK cells have several advantages, including less toxicity, a high potential opportunity for universal off-the-shelf manufacturing, increased infiltration into solid tumors, overcoming resistant tumor microenvironment, and absence of graft-versus-host disease (GVHD).

In this review, we discuss NK cell biology, the source of CAR-NK cells, CAR structure, advances, challenges, and ways to overcome these challenges in CAR-NK cell therapy. Furthermore, we have summarized and highlighted some preclinical and clinical studies in this field.

Smad4 regulates the expression of the genes required for heart homeostasis. Regarding the central role of microRNAs in cardiac biology, we investigated the expression of the three Smad4-targeting miRNAs, namely miR-18a-5p, miR-19a-3p, and miR-20a-5p, as well as Smad4 during differentiation of human endometrium-derived mesenchymal stem cells (hEMSCs) into cardiomyocytes (CMs).

To evaluate mesenchymal phenotype and multi-lineage differentiation ability of hEMSCs, immunophenotyping by flow cytometry and differentiation into osteoblasts and adipocytes were performed, respectively. For transdifferentiation into CMs, hEMSCs were exposed to a cardiomyogenic medium composed of 5-aza and bFGF for 30 days. The comparison between transcriptional expression levels of Nkx2-5, GATA4, Smad4, TNNT2, TBX5, miR-18a-5p, miR-19a-3p, and miR-20a-5p by qRT-PCR, as well as protein levels of Nkx2-5, Smad4, and cTnT by immunofluorescence staining, was conducted in every 6 days.

In vitro, the mesenchymal stem cell phenotype of hEMSCs and their potency for differentiation into other MSCs were confirmed. Differentiated hEMSCs had morphological characteristics of CMs. The percentage of positive cells for Nkx2-5, Smad4, and cTnT proteins was increased following induction and culminated on the 24th day. Also, mRNA levels of Nkx2-5, GATA4, Smad4, TNNT2, and TBX5 exhibited the same trend. The expression of investigated miRNAs was significantly decreased sequentially. A significant negative correlation between expressions of Smad4 and investigated miRNAs was observed.

Our results indicate that miR-18a-5p, miR-19a-3p, and miR-20a-5p are involved in the cardiac differentiation propensity of hEMSCs potentially by regulation of Smad levels. Although, more mechanistic experiments are required to confirm this idea.

Gastric cancer is one of the most commonly known malignancies and is the fifth cancer-related death globally. Whereas natural killer (NK) cells play a critical role in tumor elimination; therefore, adoptive NK cell therapy has become a promising approach in cancer cytotherapy. Hence, this study investigated the chemoimmune cell therapy in MKN-45 derived xenograft gastric cancer model.

Three groups of animals have received the following treatments separately: activated NK cells, capecitabine, the combination of capecitabine and activated NK cells, and one was considered as the control group. Morphometric properties of tumor samples were evaluated at the end of the study. NK cells infiltration was evaluated by immunohistochemistry (IHC) of hCD56. Mitotic count and treatment response was assessed by hematoxylin and eosin (H&E) staining. The proliferation ratio to apoptosis was determined by IHC assessment of Ki67 and caspase 3.

The results indicated that the NK cell therapy could effectively decrease the mitotic count in pathology assessment, but the tumor was not completely eradicated. In combination with metronomic chemotherapy (MC) of capecitabine, NK cell therapy demonstrated a significant difference in tumor morphometric properties compared to the control group. The proliferation ratio to apoptosis was also in line with pathology data.

Although NK cell therapy could effectively decrease the mitotic count in vivo, the obtained findings indicated lesser potency than MC despite ex vivo activation. In order to enhance NK cell therapy effectiveness, suppressive features of the tumor microenvironment and inhibitory immune checkpoints blockade should be considered.

Angiogenesis is a characteristic of glioblastoma (GBM), the most fatal and therapeutic-resistant brain tumor. Highly expressed angiogenic cytokines and proliferated microvascular system made anti-angiogenesis treatments a thoroughly plausible approach for GBM treatment. Many trials have proved to be not only as a safe but also as an effective approach in GBM retardation in a certain time window as seen in radiographic response rates; however, they have failed to implement significant improvements in clinical manifestation whether alone or in combination with radio/chemotherapy. Bevasizumab, an anti-vascular endothelial growth factor-A (VEGF-A) antibody, is the only agent that exerts meaningful clinical influence by improving progression-free survival (PFS) and partially alleviate clinical symptoms, nevertheless, it could not prolong the overall survival (OS) in patients with GBM. The data generated from phase II trials clearly revealed a correlation between elevated reperfusion, subsequent to vascular normalization induction, and improved clinical outcomes which explicitly indicates anti-angiogenesis treatments are beneficial. In order to prolong these initial benefits observed in a certain period of time after anti-angiogenesis targeting, some aspects of the therapy should be tackled: recognition of other bypass angiogenesis pathways activated following antiangiogenesis therapy, identification of probable pathways that induce insensitivity to shortage of blood supply, and classifying the patients by mapping their GBM-related gene profile as biomarkers to predict their responsiveness to therapy. Herein, the molecular basis of brain vasculature development in normal and tumoral conditions is briefly discussed and it is explained how "vascular normalization" concept opened a window to a better comprehension of some adverse effects observed in anti-angiogenesis therapy in clinical condition. Then, the most targeted angiogenesis pathways focused on ligand/receptor interactions in GBM clinical trials are reviewed. Lastly, different targeting strategies applied in anti-angiogenesis treatment are discussed.

Recently, menstrual blood-derived stem cells as a unique source of stem cells with some features such as ease of access, high ability to proliferate and regenerate, lack of immune system stimulation and no tumorigenesis have raised great hopes for heart disease cell therapy. However, the regulatory mechanisms and role of miRNAs in controlling the differentiation of stem cells into cardiomyocytes are not fully understood. In this study, the level of human miR-26b-5p microRNA were investigated before and after differentiation of endometrial stem cells into heart cardiomyocytes.

Endometrial mesenchymal stem cells were differentiated into cardiomyocyte-like cells for 30 days in the presence of 5-azacitidine and fibroblast growth factor. Then, using bioinformatics studies, human miR-26b-5p microRNA was selected and its expression pattern was performed during days 0, 6, 12, 18, 24 and 30 of differentiation by Real Time qRT PCR

Expression Level of human miR-26b-5p showed an uptrend differentiation between days 0 and 6 and then showed a significant decreasing trend differentiation from day 6 to day 18 and an uptrend again after day 18.

The non-uniformity in the expression of hsa-miR-26b-5p microRNA during the 24-day differentiation induction period indicates the existence of different messaging pathways involved in the differentiation process as well as different phases in the evolution and differentiation of cardiomyocytes. The miRNAs involved in the differentiation process and their possible role in turning off and on these messaging pathways at the beginning or end of a phase seems necessary.

Diabetes is one of the metabolic diseases characterized by hyperglycemia, with many complications. Diabetic foot ulcer (DFU) is a significant complication of diabetes. Various therapy procedures have been recently described for DFU improvement.

Using PubMed, Scopus, Science Direct, and Google Scholar to discover the therapeutic effects of bee products, this review study was conducted in 2018-2019 by searching PubMed, Scopus, Science Direct, and Google Scholar databases.

Cell therapies with various cell candidates such as mesenchymal stem cells (MSCs) are increasingly introduced into routine medical care to manage skin wounds. The applying of these cells for tissue regeneration was initially based on the capability of MSCs to differentiate into specialized cells within the injured tissue. Paracrine signaling and differentiation mechanisms have both been contributed to improving tissue repair by MCSs. However, the role of MSCs differentiation is less due to the poor survival of these cells at the site of injury.

At the same time, paracrine signaling or their secretome is the primary mechanism of MSCs that stimulate neovascularization and re-epithelialization and mobilization of inhabitant stem cells. In this review study, we discuss the role of MSCs and their secretome that can improve the use of this new approach in treating ulcers and DFU.

Neuroblastoma (NB) is one of the most prevalent childrenchr('39')s neoplasm and some of which still do not respond to common treatments and is an ideal candidate for NK cell immune therapy. In this research, we have performed cytokine activation of NKs by using proper interleukins to obtain more cytolysis which was evaluated in vitro and in vivo.

Isolation of NKs from human peripheral blood was performed using CD56 marker via magnetic activated cell sorter (MACS). The IL-2 and IL-15 cytokines were used for proliferation, and IL-21 for activation of cells. The neuroblastoma model was developed using the SK-N-SH cell line in Nude mice. The in vitro effect of the NKs on the two neuroblastoma lines were assessed after activation and via flow cytometry confirmation.

The results showed that at the ratio of 1:10 the IL-21 activated NKs lysed 94% of the SK-N-SH and 91% of the CHLA-255 cell lines (P<0.05), in spite of the only 40% and 38% of lysis for these cell lines without activation (P<0.05), respectively. The NKs activated by IL-21 were able to eliminate 70% (P<0.05) of the xenograft neuroblastoma tumors in nude mice.

The present investigation was showed that the activation of NK cells using IL-21 can be useful in the treatment of neuroblastoma and could be applied in future clinical trials.

Program death 1 (PD-1)/ program death-ligand 1 (PD-L1) pathways, as the main inhibitory checkpoints, induce immunosuppression in the tumor microenvironment (TME). Despite the importance of inhibitor checkpoint receptor (ICR) blockers, their outcomes have been limited by the low immune response rate and induced acquired resistance. Pre-existing tumor-specific T cells is related to the improvement of their therapeutic efficacy. In the present study, we show that the combination of liposomal gp100 nanovaccine with anti PD-1 monoclonal antibody (mAb) potentiates the therapeutic effect in the melanoma model.

In this study, we first decorate the cationic liposome with gp10025-33 self-antigen and then characterize it. Mice bearing B16F10 melanoma tumors were vaccinated with different formulations of gp100 peptide (free or liposomal form) with or without CpG ODN adjuvant in combination with anti PD-1 mAb.

Therapeutic combination of liposomal nanovaccine and CpG with anti PD-1 mAb, demonstrated the increased number of tumor infiltrated lymphocytes (TILs) in TME with the highest IFN-γ production and cytotoxic activity, which led to remarkable tumor regression.

Our results demonstrated the synergism between Lip-peptide+CpG nanovaccine and anti PD-1 regime, which improved the therapeutic efficacy of PD-1 checkpoint blocker in melanoma mice models.

Endometrium is recently introduced as an available source of mesenchymal stem cells (EnMSCs), which can be obtained without anesthesia and side effects. Regarding the issues and complexities of cell-based therapies, exosomes gain tremendous attention as a novel tool for cell-free therapies. Although several clinical trials are recently established based on therapeutic potential of EnMSCs, biological roles of EnMSC-derived exosomes are still unclear.

The current study was conducted to investigate the potential effects of EnMSC- derived exosomes on proliferation, migration, and angiogenesis of human umbilical cord vein endothelial cells (HUVECs). For this purpose, EnMSCs and then EnMSC-derived exosomes were isolated and characterized. MTT assay and wound healing assay as well as tube formation assay were applied.

The collected data showed that EnMSC-derived exosomes significantly increased proliferation, migration, and angiogenesis of HUVECs. It was observed that the effects of exosomes were applied in a dose dependent manner. In addition, expression analysis by quantitative real-time PCR showed that increased expression of proliferation and angiogenesis genes in HUVECs were treated with EnMSC-derived exosomes in a dose dependent manner.

The current study results showed that EnMSC-derived exosomes can exert biological effects such as their source cells and become new candidates for cell-free therapies. Taken together, increased angiogenesis makes EnMSC-derived exosomes a promising tool in regenerative medicine, especially wound healing and treatment of vascular disease

Fluoxetine is a common antidepressant which selectively inhibits serotonin reuptake at synaptic level. Some research findings have proposed the effect this drug on neurogenesis, neuronal survival as well as proliferation of the neural progenitor cells. Endometrium is a part of uterus which harbors mesenchymal stem cells. This source of stem cells can be differentiated into neural cells which may potentially be used in treating many diseases. Given the above, this study was designed to assess the effect of fluoxetine on neural cells differentiation from endometrial stem cells. Endometrial stem cells obtained from stem cell banking were cultured in Dulbecco’s modified eagle’s medium (DMED) containing fetal bovine serum (FBS) in the presence of Retinoic Acid, fluoxetine and Retinoic Acid+fluoxetine for 10 days. To assess the differentiation of endometrial stem cells into neural-like cells, we used immunocytochemistry and RT-PCR. The viability of cells was assessed using the trypan blue test. Data analysis revealed that 61% of endometrial stem cells differentiated to neural-like cells. Moreover, the biopotency of neural-like cells on fluoxetine treatment was more pronounced across differentiation days. Based on our findings, fluoxetine was shown to be a suitable inducer for the differentiation of neural-like cells from endometrial stem cells.

Due to increasing clinical demand for adipose tissue, a suitable cell for reconstructive adipose tissue constructs is needed. In this study, we investigated the ability of Human Endometrial-derived stem cells (EnSCs) as a new source of mesenchymal stem cells to differentiate into adipocytes. EnSCs are the abundant and easy available source with no immunological response, for cell replacement therapy. Single-cell suspensions of EnSCs were obtained from endometrial tissues from 10 women experiencing normal menstrual cycles, and were cultured at clonal density (10 cells/cm2) or limiting dilution. Endometrial mesenchymal stem cell markers were examined flow cytometry. These cells were treated with adipogenic-inducing medium for 28 days. The adipogenic differentiation of the EnSC was assessed by cellular morphology and further confirmed by Oil Red O staining and RT-PCR. The BM-MSC differentiated into adipocytes in the presence of adipogenic stimuli for 3 weeks. The flow cytometric analysis showed that the cells were positive for CD90, CD105, CD146 and were negative for CD31, CD34.We showed that the key adipocytes marker PPARa was expressed in mRNA level after 28 days post treatment (PT). According to our finding, it can be concluded that EnSCs represent a useful in vitro model for human adipogenesis, and provide opportunities to study the stages prior to commitment to the adipocyte lineage.

Zinc is an essential micronutrient for growth and proper immune function. Zinc deficiency may contribute to the incidence, prevalence and severity of diarrhea and can also cause failure to thrive in children. The purpose of this study was to evaluate the serum zinc level in children with diarrhea.

This cross-sectional study was performed on 105 children with acute watery diarrhea referred to Kashan Shahid Beheshti hospital during 2008-9. To test the serum zink level, a sample of venous blood (5-cc) was taken. In addition, the age, sex, the duration of diarrhea, the length of hospitalization and FTT were evaluated. Data were analyzed using Chi-square, T-test and ANOVA.

Among the 105 children, 48.6% were male and half of them aged more than 12 months. The results showed that the mean serum zinc level in hospitalized children and diarrhea duration more than 3 days was lower than the other children (P=0.023 and P=0.004, respectively). Moreover, the mean serum zinc level in children with FTT was lower than the children without FTT (P<0.001).

Duration of diarrhea, the length of hospitalization and FTT in children with low serum zinc levels was more than the other children.

Nosocomial infections is a common source of infection in hospitalized patients. Prior reports have shown the possibility of venous catheter infection, too. The present study was carried out to determine the venous catheter infection of patients admitted to ICU and its related factors in Shaheed Beheshti hospital in Kashan in 2000. It was descriptive study. Patients for whom IV catheters were removed were included. Then the 3cm of the catheters were cut aseptically and put in Thayer-glycolate. In case of growth within the next 7 days, the sample would be passage on agar. Finally the type of cultured bacteria as well as its antibiogram was determined. Infectious related factors were considered as: Age, sex, duration of catheter usage, duration of ICU hospitalization, catheter-induced phlebitis and the location of catheter. Of 100 cultured catheter removed from 36 patients, 29% revealed to have infection, among which, staphylococcus coagulase-negative was the most prevalent (18%). Age, duration of catheter usage, duration of ICU hospitalization, catheter-induced phlebitis and the location of catheter had no effects on infection rate (NS). Staphylococcus coagulase-negative is the skin normal flora and do not cause severe infection, thus the true rate of infection is less. Needless to say, the venous catheter infection is a critical problem seeks further attention in hospitalized patients.