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COVID-19, an infectious viral disease caused by severe acute respiratory syndrome (SARS-CoV-2) with more than 260 million infections as of December 2019, is a serious threat to the health and economy of human societies. Designing and producing a suitable vaccine can Reduce the incidence of this disease. Therefore, it is very important to find effective and safe neutralizing antibodies and vaccines for COVID-19. The RDB section of the spike protein is a suitable option for the production of subunit vaccines and neutralizing antibodies. This research aims to introduce the RBD section as a vaccine candidate.

The RBD was recombinantly expressed in two hosts, Escherichia coli (E.coli) and insect cells, and its production rate was compared. Using the western blotting method, protein production was confirmed. To evaluate the function of the protein expressed in two prokaryotic and eukaryotic hosts, using the serum of patients recovered from COVID-19 (wild type and delta), the ELISA method was used.

Despite the higher production of recombinant protein by E.coli, the affinity of the antibodies in the serum of the patients to the protein expressed in the insect cell was higher.

In this study, the RBD was expressed in two different hosts. The results show that RBD is mentioned as a vaccine candidate. Homozygous in the insect cell preserves the biological activity of the protein by carrying out the process of post-translational changes such as glycosylation.

Given the global epidemic of COVID-19, it is important to design a vaccine for prevention. The virus belongs to the beta-coronavirus family and forms appendages on the surface of the glycoprotein spike virus membrane. Studies on SARS-CoV-1 and related MERS-CoV vaccines have shown that the spike protein on the virus surface is a suitable target for the vaccine. In this experimental study, we compare the recombinant fragment of spike protein (rfsp) expressed in the eukaryotic host CHO-K1 cell and prokaryotic E. coli in terms of immunogenicity, neutralizing activity, and epitopes recognition Similar to the virus strain and the ability to bind to the serum of improved patients, in two types of alpha and delta variants. The results showed that both rfSP proteins are a potential new antigen candidate for the development of the Covid 19 vaccine, but the CHO cell maintains the biological activity of the protein by performing post-translational modification processes such as glycolysis. This increases the present likelihood of developing virus-like epitopes and increases the titer of rfsp-specific antibodies in the serum of immunized mice. Therefore, priority is given to rfsp expressed in CHO cells to evaluate vaccine efficacy.

PD-L1, one of the most important immune-regulating molecules in all cancers including breast cancer, suppresses the immune system against tumor cells. MiRNAs control PD-L1 expression as therapeutic tools and targets. This study investigated the role of miR-200c in reducing PD-L1 expression and breast cancer cell proliferation.

In this study, miR-200c targeting the PD-L1 3'UTR was confirmed by bioinformatics and dual-luciferase assay. PD-L1 expression in breast cancer tissues and cell lines was checked compared to normal tissues and cell lines. qRT-PCR was used to measure PD-L1 expression after miR-200c transfection into breast cancer cell line. The MTT test was used to determine the effect of miR-200c introduction on cell proliferation. 

The 3'UTR region of the PD-L1 gene was identified as a target for miR-200c using bioinformatics software and a dual luciferase assay. The decrease in miR-200c expression in breast cancer cells is directly related to the increase in PD-L1 expression (r =-0.68, P value₌ 0.0158). Also, increasing the expression of miR-200c in MDA-MB231, BT549, and MCF7 breast cancer cells decreased the expression of PD-L1 by (0.217±2.3, 0.021±2.11, and 0.012±0.146, respectively). The increased expression of miR-200c decreased the proliferation rate in MDA-MB231 by 0.45 ± 0.012, BT549 by 0.256 ± 0.2, and in MCF7 by 0.164 ± 0.02, compared to non-transfected cells. (Mean ± SE)

MiR-200c may be able to prevent breast cancer progression by targeting the PD-1/PD-L1 pathway, reducing PD-L1 expression, and reducing breast cancer cell proliferation.

Kidney diseases are an important medical problem worldwide. Since there are limited treatment options for damaged kidneys, stem cell therapy has become an alternative treatment. The aim of present study is to investigate effect of culture medium obtained from mesenchymal stem cells (MSCs-CM) of newborn mice kidneys in percentages of 10, 30 and 50 on differentiation of embryonic stem cells towards kidney epithelial cells. Mesenchymal stem cells were isolated from kidneys of newborn mice, passaged and propagated. Determination of the identity of cells was done using flow cytometry and checking the expression of surface markers CD105, CD29, CD90. In third passage of extracted cells, supernatant culture medium was collected, hESC were cultured and multiplied in the complete culture medium of cells and the differentiation of hES cells into progenitor cells was investigated. The expression of PAX2, ZO1 and CK18 genes was investigated using RT-PCR, expression of CD133, CD24 and CD44 surface markers was investigated using flow cytometry. Flow cytometry results confirmed the mesenchymal nature of the cells. The results of differentiation of hESCs showed that expression of PAX2, ZO1 and CK18 genes increased significantly (p<0.05) in the groups containing supernatant. The results of flow cytometry show an increase in expression of CD133 and CD24 markers in groups containing CM and the expression of CD44 marker in the group containing 50% CM, compared to control group. In general, results showed supernatant culture process of cells has a positive effect on inducing differentiation of human embryonic stem cells into kidney progenitor cells.

 Defensive barrier is important in preventing infection and disease progression caused by E.coli O157:H7. Among beneficial effects of probiotics, immune system modulation by producing cytokines is one of the most common reported benefits. The aim of this study was to investigate the Lactobacillus acidophilus probiotic effect on il10 cytokine gene expression in Zebrafish infected with E. coli O157:H7 as a laboratory model.

 The 300 pieces of fish were grouped equally; control group (C1A1) received basic diet, group (B1A1) received basic diet and faced E.coli O157:H7, treatment 1 group (T1A1) received basic diet containing L. acidophilus and no exposure to E. coli O157:H7, treatment 2 group (T2A2) received basic diet containing L. acidophilus and exposed to E. coli O157:H7. During the 30-day period of testing, intestinal tissue samples were taken on days 15, 27 and 30 for il10 gene expression and bacterial colonization studies.

 The findings indicated bacterial colonization in fish intestines. Also, the expression of il10 gene in the experimental groups decreased after exposure to E. coli O157:H7 (P<0.01). This decrement was less with the regulation of il10 in the groups fed with L. acidophilus compared to the control group. In Zebrafish fed with L. acidophilus, the expression of the il10 gene increased gradually (day15, day27 and 30; P<0. 01).

 Overall, the results showed that the L. acidophilus was able to increase il10 gene expression, boost the immune system in Zebrafish as a laboratory model. Therefore, L. acidophilus La5 probiotic can be used as a possible useful candidate against E. coli O157:H7.

Background &

In today's societies, breast cancer has significantly threatened women's health (1). Successful cancer treatment can also destroy tumor germ cells (2, 3). Treatment methods based on immune checkpoint molecules are considered (4). PD-L1 overexpression suppresses the immune response to cancer cells and causes epithelial-to-mesenchymal transition (EMT) and cancer stem cell (CSC) phenotypes, metastasis and chemical resistance (5). Overexpression of PD-L1 and MUC1 is observed in approximately 90 TNBCs. In this present study, we investigated the inhibitory effect of miR-143-3p on MUC1 expression as an oncogene that has the most impact on PDL-1 overexpression in BC cell lines MCF-7 (less aggressive BC), MDA‐MB‐231, and BT-549 (metastatic BC model). The suppressive effect of miR-143-3p on the expression of this oncogene as a direct effect and PDL1 level as an indirect result in BC cell lines was investigated. Besides, the effect of miR-143-3p overexpression on BC cell lines proliferation through downregulation of candidate oncogene was investigated.

First, the targeting of miR-143-3P on MUC1 3 'UTR was confirmed by using bioinformatics software and then by dual luciferase assay. The expression level of miR-143-3P and MUC1 were measured in breast cancer lines compared to the normal line. After transfection of miR-143-3P into breast cancer cell lines, MUC1 and PD-L1 expression in mRNA levels were evaluated by using qRT-PCR. The miR-143-3p effects on MUC1 and PD-L1 expression and its impact on cell proliferation was investigated by using MTT assay, (mean ± SD).

The result of the dual luciferase assay was obtained from cloning the 3'UTR of the MUC1 gene in the psi-check2 vector and sequencing. According to the prediction analysis, the 3′-UTR of MUC1 gene has been considered as a putative miR-143-3p binding site (Target scan database). Transfecting miR-143-3p into MDA-MB-231, BT-549, MCF-7 cells harboring MUC1 3′-UTR decreased the luciferase activity to to 55% ± 1/73, 57% ± 1 /15 , 59 % ± 3/51% respectively, in comparison with the control (P < 0.01) (mean ± SD). he expression of miR-143-3p was significantly downregulated in BC cell lines compared to their control, and its downregulation was negatively correlated with MUC1 overexpression (r= -0.6419, p = 0.0244). Increased expression of miR-143-3P, decreased the mRNA level of MUC1 in MDA-MB231 to 1/3 ± 0/7, BT549 to 1/1± 0/45 and in MCF7 to 0/4/0 ±21, compared whit untransfected cells.
MiR-143-3p introduction to BC cells downregulates PDL-1 indirectly. Overexpression of miR-143-3P reduces the MUC1expression, and the reduction of MUC1 expression reduces PD-L1 expression mRNA level in MDA-MB231 to 3/4±0.45, BT549 to 3/1±0.56, MCF7 to 0/53±0.48, compared whit un transfected cells (fig4A). miR-143-3p inhibits BC cellular proliferation. An MTT assay revealed that miR-143-3p overexpression significantly suppressed the proliferation in MCF-7 to 52% ± 1.55, BT549 to 48% ± 2.45 MDA-MB231 to 43% ± 3.36, compared whit non-transfected cells 72 h after transfection.

 Breast cancer treatment has made significant progress in recent years, and a new treatment called immunotherapy, which uses the body's immune system to treat cancer, has become popular (6). One of the symptoms of breast cancer tumors is the overexpression of MUC1. A high expression level of MUC1 is associated with poor prognosis in cancer patients (7, 8). In an animal model of breast cancer, intravenous injection of MUC1 suppresses the immune system and accelerates the death of tumor-bearing mice. The serum level of MUCl expression is also associated with suppression of the immune system in patients with metastatic adenocarcinoma treated with active specific immunotherapy (8, 9). As we mentioned earlier, MUC1 can increase the expression of PD-L1 by recruiting MYC and NF -kB p65 to the PD-L1 promoter in some solid tumors, such as triple negative breast cancer (10). the mechanisms underlying PD-L1 expression in MUC1-positive tumor tissue were investigated, and the potential of targeting PD-L1 to inhibit the progression of MUC1-overexpressing colon cancer in mice, Therefore, the role of MUC1 in tumor immune evasion has been confirmed and can be a basis for using PD-L1 inhibitors in MUC1-positive colon cancer (8). microRNAs can regulate the expression of immune checkpoint receptors and their ligands (11). Changes in the expression of miR-143 often play a role in tumorigenesis (12). In 2020, Y Tokumaru and his colleagues showed by studying breast cancer samples with estrogen receptors that the high expression of microRNA-143 is associated with a favorable tumor immune microenvironment and better survival in breast cancer. They reported that miR-143 is a tumor suppressor microRNA that exhibits anti-tumor function by targeting KRAS signalling pathways in various malignancies. The high expression of miR-143 in cancer cells with a favorable tumor immune microenvironment regulates the positivity of anti-cancer immune cells. The suppression of pre-cancerous immune cells is related and leads to better survival of breast cancer patients (13).
In this study, we demonstrated MUC1 was potential as a novel target of miR-143-3p, therefore, the dual-luciferase reporter assay verified that miR-143-3p directly targeted MUC1 and inhibited this transcription. We found miR-143-3p downregulated in breast cancer cell lines compared to the normal line via qRT-PCR. In the current study, the qRT-PCR results illustrated an inverse relation between MUC1 mRNA expression and miR-143-3p expression in breast cancer cell lines. We investigated indirect targeting of PD-L1 via MUC1 targeting by miR-143-3p., be able to be prevent the cell proliferation in breast cancer cell line. Therefore we recognized, targeting MUC1 may be a novel strategy for BC therapy and prevent the progression of breast cancer and miR-143-3p as a promising candidate for miR restoration therapy in BC patients.

Genome editing is an efficient and accurate method in biological and medical studies. Among the wide range of genome editing techniques, clustered regularly interspaced short palindromic repeats (CRISPR) is one of the simplest and yet promising methods. The CRISPR system consists of two key components; an endonuclease enzyme called Cas9 and guide RNA (gRNA), ensuring that Cas9 enzyme cuts at the right point in the genome. Although CRISPR genome editing is one of the useful methods to modify the genome, there are still multiple challenges in the technique steps.

sgRNAs were designed and ordered accordingly on the basis of the target site and vector of interest. Cloning procedures were performed and confirmed by sequencing as a gold standard. The list of high efficient sgRNAs for the LAMP-2 gene was prepared based on the available outstanding databases by analyzing and comparing the specificity and functionality as two main characteristics.

In this study, we tried to evaluate the main challenges in the process of preparing CRISPR/Cas9 popular vectors (pX330/pX459). A list of high efficient sgRNAs for an autophagy gene is also prepared as an example for clarification of the sgRNA design procedures.

This study tried to depict problems that may be encountered during sgRNA design and plasmid preparation, followed by giving appropriate recommendations.

The protective role of astaxanthin on human health, including the prevention of cardiovascular disease, prevention of cell aging, refers to the very strong antioxidant properties of this substance. Cadmium is one of the pollutants in ecosystems. This heavy metal acts as a toxin on the reproductive system and destroys the process of spermatogenesis in testicular tissue. The aim of this study was to evaluate the effect of astaxanthin on cadmium chloride induced testicular damage in adult male Wistar rats. In this study, 45 rats were randomly divided into 9 groups. The healthy control group did not receive any treatment. Control group, physiological serum, infertile control group, cadmium chloride (1 mg / kg body weight) by intraperitoneal injection, astaxanthin receiving groups (5, 10 and 20 mg / kg body weight) by gavage and groups They received cadmium chloride and astaxanthin (5, 10 and 20 mg / kg) at the same time. Test samples from testicular tissue homogenates were used to measure oxidative stress parameters. The parameters were analyzed by photometric method with ELISA reader and microplate reader. The results showed that astaxanthin (10 and 20 mg / kg body weight) increased the antioxidant enzymes SOD, GPX and CAT and astaxanthin (10 mg / kg body weight) decreased MDA. Thus, astaxanthin may act as an antioxidant in protecting the rat testis against oxidative stress induced by cadmium chloride.

Iran is one of the most important hot spots for cutaneous leishmaniasis (CL) in the world. To date, no studies have been done on both epidemiological aspects along with the length of the treatment course of CL. This study aimed to determine the relative frequency of CL in patients with suspected skin lesions and the duration of healing after treatment with different regimens.

This cross-sectional study was conducted on patients with CL referred to the skin diseases and leishmaniasis research center (SDLRC) in Isfahan during the years 2018 to 2019. Among 389 patients with suspected skin lesions, 150 cases were included with proven CL. Information such as age, sex, education, location, size of the lesion, duration of treatment, and the rate of recovery were recorded. SPSS software version 20 was used for data analysis, the chi-square, Fisher´s Exact, and one Way ANOVA tests were used with a significant level of p < 0.05.

Among 350 admitted cases, 150 cases were CL. positive (42.85%). The rate of complete recovery was higher in cases with an average age of 33.55 ±18.9 years, but these differences were not statistically significant (P =0.077). There was 34 cases more than the other groups in this range of age. ( The rate of complete recovery in patients with a history of migration to endemic areas was higher than in patients without a history of migration (P = 0.81)). The rate of complete recovery in patients whose means treatment duration was 59.03 ± 41.43 days was higher than other recovery periods (P = 0.23).

The rate of complete recovery was higher in adult cases than the other groups. In this study, it was proved that the rate of recovery of patients had the significant relationship with the average duration of treatment.

Akkermansia muciniphila, an important member of the gastrointestinal microbiota, is involved in the functioning of the host immune system. We aimed to investigate the effect of A. muciniphila and its OMVs on the expression of microRNA-21 and microRNA-34a involved in the maturation of human dendritic cells. A. muciniphila was cultured on a mucin-containing medium and its OMVs were extracted by ultracentrifugation. Extraction of peripheral blood mononuclear cells (PBMCs) was performed using Ficoll’s density gradient centrifugation method. Monocytes were differentiated into dendritic cells in the presence of interleukin-4 cytokines and granulocyte-monocyte colony-stimulating factor. Then, A. muciniphila at MOI 50 and 100, OMVs at a dose of 50 µg/µl, dexamethasone (to induce tolerogenic dendritic cells), and the LPS (to induce mature dendritic cells) were incubated. Finally, the expression of microRNA-21 and microRNA-34a was measured by real-time PCR. The morphological characteristics of the extracted OMVs showed that their size was between 20-200 nm. In LPS treatment, there were longer cytoplasmic growths than OMVs of dexamethasone and A. muciniphila groups. MicroRNA-21 expression was decreased in the LPS group compared to dexamethasone, A. muciniphila, and OMV groups, although this decrease was not significant (P> 0.05). In addition, microRNA-34a expression was significantly increased in the LPS group, which was reversed by treatment with dexamethasone, A. muciniphila, and OMV (P <0.001). MicroRNA-21 and microRNA-34a can exhibit anti-inflammatory and pro-inflammatory functions. A. muciniphila and its OMVs appear to be a viable option for introduction as a new generation probiotic.

Consuming a high-fat diet, as a stressor, during pregnancy and lactation can change the programming of the neuroendocrine system, and cause long -lasting adverse metabolic and behavioral effects in later life. In this study, the effect of high-fat diet feeding during pregnancy-lactation on basal plasma corticosterone level in adult male rat offspring was investigated. This study was performed in 1398, at Physiology Department, Shahid Beheshti University of Medical Sciences.

In this experimental study, dams received high-fat and normal diets from the beginning of pregnancy to the end of lactation. At the end of weaning, male offspring were divided into high-fat diet (HFD) and normal diet (ND) (6 rats per group according to previous studies) based on the dams’ diet and received normal diet until six weeks of age. Then the body weight of the animals was measured using a digital balance and blood samples were taken from their tails to determine the plasma concentration of corticosterone by a rat corticosterone Elisa Kit. Subsequently, their adrenal glands removed and weighed using a digital balance. The results were analyzed using independent t test.

Compared with the offspring of the ND group, the plasma concentration of corticosterone was higher in the offspring of the HFD group (2.11±0.13) (P<0.01), whereas the body weight of these offspring (114.8±2.37) (P<0.01) was lower. However, the adrenal gland weight of HFD group (28.25±2.80) was not significantly different from that of ND group (29.52±1.68). There were not any reduction in the samples of the study groups.

  Regarding the unchanged adrenal glands weight of the HFD group, it seems that the increase in plasma concentrations of corticosterone in this group is due to impaired corticosterone clearance.

Multiple sclerosis (MS) is one of the most common diseases of the nervous system, characterized by inflammation of the central nervous system and oxidative stress. Carvacrol is a monoterpenoid phenol with antioxidant effects against free radical. The aim of this study was to evaluate the effect of carvacrol on the expression of Hmox-1, iNOS, Nrf2 and NF-ҚB genes in the spinal cord of Experimental Autoimmune Encephalomyelitis (EAE) rats as an animal model of MS.

EAE was induced in female Lewis rats and they were then divided into three groups: control, EAE model, and EAE treated with carvacrol. Carvacrol was daily injected intraperitoneally for 17 days after immunization. RNA was extracted from rat spinal cord tissue and changes in the expression of the Hmox-1, iNOS, Nrf2 and NF-ҚB genes were examined by Real-Time PCR.

The results showed that carvacrol treatment stopped severe weight loss in the EAE model group and their mean weight increased. Furthermore, carvacrol was found to reduce the expression of iNOS and NF-ҚB genes and increase the levels of Hmox-1 mRNA and Nrf2 mRNA.

The effect of carvacrol on weight changes in treated rats, along with the increased HO-1 gene expression in their spinal cord, associated with increased Nrf2 mRNA and decreased expression of iNOS and NF-ҚB, led to a reduction in inflammation and oxidative stress, and an increase in the neuroprotection. Furthermore, the invasive environment created for some cells, such as oligodendrocytes, was modulated.

PD-L1 is one of the most important immune control molecules in breast cancer and plays an important role in suppressing the immune system against tumor cells. Controlling the expression of PD-L1 at mRNA level using miRNA inhibitors could be helpful strategy for cancer treatment. In this study, considering the possible role of miR-145 as a tumor suppression in breast cancer, its involvement in reducing PD-L1 expression in breast cancer cell lines has been investigated.

First, the targeting of miRNA-145 on 3 'UTR of PD-L1 gene was confirmed using bioinformatics software and then by luciferase dual reporter assay. The expression level of miRNA-145 was measured in breast cancer cell lines compared to normal line. After transfection of miRNA-145 into breast cancer cell lines, qRT-PCR was used to evaluate the effect of miRNA-145 on PD-L1 expression.

we showed that decreased expression of miRNA-145 in breast cancer cell lines was directly related to increased PD-L1 expression (r= -0.6175, P value₌0.0457). In addition, increased expression of miRNA-145 in breast cancer cell lines MDA-MB231, BT549 and MCF7 significantly reduced PD-L1 expression (1.938±0.212, 1.784±0.03 and 0.083±0.02 respectively).

Our findings suggest that miRNA-145, by targeting the PD1/PD-L1 pathway and reducing PD-L1 expression, may be a therapeutic agent to prevent the progression of breast cancer.

Transforming growth factor-beta-induced factor X-linked (TGIF2LX) as a homeodomain protein and TGF-βcorepressor, could regulate proliferation of some cancer cells including colorectal cancer by some signaling pathways. Small non-coding RNAs (microRNA; miRNA) are known as molecular regulators of colorectal cancer that are involved in the processes of cell growth, proliferation, differentiation and apoptosis. The aim of this study was to evaluate the biological significance of TGIF2LX protein and its effect on the expression of oncogenic miRNAs miR-34a, miR-20a and miR-21 in colorectal cancer cells SW1116.

Human SW1116 cell line and cell line transfected with cDNA encoding TGIF2LX gene were cultured in RPMI 1640 medium under appropriate conditions. MTT assay was used to assess cell viability in vitro. After RNA extraction from all cell groups and cDNA synthesis, miRNA expression analysis was performed using qReal-time PCR technique.

The results showed that the increased expression of TGIF2LX could reduce the proliferation of SW1116 cell line. Gene expression analysis showed that increased expression of TGIF2LX could significantly reduce the expression level of miR-21 (P<0.038). However, the expression levels of miR-34a and miR-20a in SW1116 cells transfected with TGIF2LX did not show a significant change compared to non-transfected cells (P>0.05).

Our findings provide evidence of molecular mechanisms that the homeodomain protein TGIF2LX can act as a tumor suppressor in colorectal cancer cells by reducing miR-21 expression. Therefore, this protein can potentially be considered as a promising option for gene-based therapeutic strategies in this cancer.

Human Papillomaviruses (HPV) infection was known to be the leading cause of cervical cancer. Cervical cancer is the fourth most common cancer among women in the world. Designing a DNA vaccine with therapeutic properties as well as prevention can have a significant impact on reducing morbidity and mortality.

Gene sequences including the immunogenic and conserved epitopes of L1 and E7 proteins of high risk papillomaviruses were designed. After synthesis of the L1-E7 fusion construct in pUC57 cloning vector, its subcloning was performed in pcDNA3.1 (-) eukaryotic vector. The concentration and purity of the recombinant pcDNA-L1E7 plasmid was determined by NanoDrop spectrophotometry.  

The gene sequence after cutting from pUC57-L1E7 by BamHI/HindIII restriction enzymes, was subcloned in the same cloning site of pcDNA3.1 (-) vector. The presence of gene was confirmed by digestion with BamHI/HindIII enzymes, as a 639 bp fragment on 1% agarose gel. The recombinant pcDNA-L1E7 plasmid was purified by endotoxin-free extraction kit for isolation of bacterial lipopolysaccharide. The concentration of the purified DNA was obtained about 1474 ng/ µl.

Cloning of the L1-E7 fusion construct in the eukaryotic vector was successful. In the next steps, the recombinant vector will be used for gene vaccine studies in vivo.

Lung cancer is the most common malignancy with the highest mortality worldwide. LncRNAs play complex roles in the initiation and progression of lung cancer, thereby affecting the prognosis of patients. In this study, the expression changes of CCAT1, CCHE1, ANRIL, BANCR, and PICART1 lncRNAs in tumor tissues compared to normal tissues adjacent to the tumor were investigated and their potential as biomarkers for the diagnosis and prognosis of lung cancer was evaluated. In order to conduct this case-control study, 30 samples of lung cancer tissue and 30 samples of adjacent non-cancerous tissues (ANCTs) were collected and after total RNA extraction and cDNA synthesis, the expression levels of target lncRNAs and β2M reference gene was measured using SYBR Green real-time PCR assay. The statistical analysis of Real-time PCR data showed no significant difference between the expression of these lncRNAs in tumor tissue and normal tissue adjacent to the tumor (P> 0.05). Furthermore, no significant association was observed between the expression level of these lncRNAs and any of the demographic and pathological characteristics of patients (P> 0.05). Based on the findings of this study, it can be concluded that these lncRNAs play no role in lung cancer in Iranian patients. However, to confirm our results, it is necessary to evaluate the expression of these lncRNAs in a larger number of samples..

Bacteria naturally secret nano-scale vesicles containing a wide range of biomolecules, such as proteins, DNA, and RNA. These vesicles are called extracellular vesicles (EVs). EVs play important roles in host-microbiota interactions. For isolating EVs, different methods have been proposed and each method has its advantages and also limitations. Therefore, in the current study, efficacy of two methods used for extraction of EVs was investigated.

For this purpose, Bifidobacterium bifidum was cultured in MRS broth under anaerobic conditions. In the first isolation protocol, ultra-centrifugation was used (Ultra-method) and in the second protocol, ultra-centrifugation (Non-Ultra method) was not used. After isolation, protein content was measured by the NanoDrop system. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) technique was utilized to compare protein pattern of the EVs. Scanning electron microscopy (SEM) images of the EVs҆ samples were taken and size of the EVs was evaluated by Digimizer software.

The results showed that the EVs isolated by the Ultra-method had significantly higher vesicle-associated protein content compared to those isolated by the Non-Ultra method (3.42 and 0.26 mg/ml, respectively). More and larger EVs (up to 235 nm and with frequent size ranging between 100 – 125 nm) were isolated by the Ultra-method compared to the Non-Ultra method (up to 117 nm and with frequent size ranging between 50–75 nm). Also, protein patterns of the EVs were similar in both methods and protein bands were observed at 25 to 250 KDa in both methods.

Our results showed that ultra-centrifugation method is a more proper method for isolation of B. bifidum-derived EVs and produces a higher amount of EVs with higher protein content and proper sizes. However, further studies are required to confirm our results.

Renal ischemia-reperfusion can lead to acute renal failure. The use of antioxidants is a good way to reduce the damage caused by renal ischemia-reperfusion. Therefore, the aim of this study was to investigate the protective effect of indole-3-acetic acid on renal ischemia-reperfusion injury through its antioxidant properties in a rat model.

Forty male rats were treated for 2 weeks in four groups included (control group, ischemia-reperfusion control group, ischemia-reperfusion group with 40 mg/kg indole acetic acid treatment and ischemia-reperfusion group with 60 mg/kg indole acetic acid treatment). Then, the blood sample was taken from the heart. Serum urea and creatinine levels were measured using biochemical kits and superoxide dismutase enzyme levels were measured using photometric methods.

The results showed that ischemia-reperfusion caused a significant increase in urea )P≤0/05) and creatinine) P≤0/05) levels and a significant decrease in the level of superoxide dismutase in ischemia-reperfusion rats compared to the control group. However, indole acetic acid caused a significant decrease in serum urea) P≤0/05) and creatinine e)P≤0/05) levels and a non-significant increase in superoxide dismutase enzyme compared to the ischemia-reperfusion control group )P>0/05).

The use of indol acetic acid was involved in reducing the renal biochemical parameters of urea and creatinine, which were increased due to ischemia-reperfusion injury, and caused the level of Super oxide dismutase enzyme to increase non-significant.

The current study aims to utilize varicocele induction in male rat models to evaluate effect and impact of varicocele on DNA integrity.

66 male Wistar rats were divided into 3 groups: control, sham and varicocele-induced (VI). 2 months post-VI, Sperm parameters, percentage of sperm DNA damage, persistent histone and lipid peroxidation were assessed.

Animal weight and their epididymal length shown no significant changes but there were significant differences in testis volume and sperm parameters between VI group versus control and sham groups. Similarly, increased level of DNA damage, persistent histone and lipid peroxidation were evaluated, and compared between varicocele and control group.

Based on literature and the result of this study, it is likely that increase oxidative stress following varicocele induction leads to lipide peroxidation, DNA damages and reduced sperm parameters. In the current study, by taking the advantage of homogenous genetic background and high number of animals, we showed that varicocele have damaging effect on DNA and chromatin integrity and should be reversed in human before aiming for pregnancy.

There is a circadian rhythm and diurnal variation in the redox capacity in blood plasma and there is a decrease in antioxidant capacity (TAC) during the RBCs storage in blood banking. So it is possible that donated blood, based on donation time, has a different antioxidant capacity and as a result a different biochemical response during storage. This study attempts to uncover this issue.
 

In this experimental study, Twenty RBC units were collected from two donor groups in the morning (8-11am) and evening (5-8pm) and were stored for 3 to 42 days at 4 to 8 degrees Celsius. Glucose, Sodium, potassium, lactate, lactate dehydrogenase (LDH), pH, malondialdehyde (MDA), and total antioxidant capacity were measured by a commercial kit. Nitrate/nitrite was measured by Griess method.
 

The levels of nitrate/nitrite, MDA, and LDH in the evening group increased significantly when compared to the morning group (p < 0.05). However, a decreasing trend in TAC, pH, Glucose, and sodium levels (p < 0.05) versus an increasing trend in potassium, lactate, LDH, and MDA was seen in both groups during storage (p < 0.05).

This study showed that oxidative damage in donated blood in the evening was higher than in the morning during storage, which may reduce RBC survival. Therefore, more studies are required with more sample sizes to examine the effect of donation time on the RBC storage lesion.

Background and Objectives One of the most important mechanisms of hematologic malignancies is DNA methylation in the promoter region of tumor suppressor genes such as P15. Harmalin is one of the alkaloids derived from plants that show anti-proliferative activity on leukemic cell lines. The effect of harmalin on Dnmt1 gene expression and hypomethylation of P15 promoter in NB4 cell line was evaluated.
Materials and Methods In this experimental study, cytotoxic effects of harmaline were investigated until 72 hours on the NB4 cell line using MTT. Flow cytometry was performed in order to evaluate the cell cycle of NB4 cell line after treatment with harmalin. Gene promoter methylation status of P15 was evaluated by Methylation Specific PCR. Dnmt1 and P15 gene expression was analyzed by Real-time PCR technique.
Results 15μg/ml harmalin after 48 hours showed a dose and time-dependent anti-proliferative properties on NB4 cell line, cell cycle analysis would stop NB4 cells in G0G1 phase of the cell cycle. Real-time PCR results showed that 15μg/ml harmalin reduced gene expression of Dnmt1, induced hypomethylation of P15 gene promoter and increased P15 gene expression in NB4 cell line.
Conclusions According to our data, the effect of harmalin on reduction of Dnmt1 expression, P15 promoter hypomethylation and P15 reactivation in this cell line showed that it could be suggested as a therapeutic strategy, suitable for monotherapy or supplement for medication which is commonly used for APL (Acute Promyeloid Leukemia).

Mesenchymal Stem Cells (MSCs) are well known as the regulator of the immune system. These multipotent non-hematopoietic progenitor cells have been originally isolated from bone marrow, and later on found in several other tissues, such as skeletal muscle, umbilical cord blood, adipose and fetal liver tissues. Immunomodulatory effects of MSCs on a variety of immune cells such as T and B lymphocytes, Natural Killer cells (NK), neutrophils, macrophages and dendritic cells, has made a good candidate of them for the treatment of inflammatory disorders, particularly autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. In addition, several studies have indicated mechanisms by which MSCs could reduce immune cell proliferation and activation leading to immune tolerance induction. Since T lymphocytes are considered as the most important immune cells, effect of MSCs on the activity of these cells has a very special significance to direct immune response. Under various conditions, T-lymphocytes have different phenotype and performance and can be differentiated into particular subtype such as regulatory T cells. Both in vitro and in vivo studies have indicated that MSCs modulate innate and adaptive immune system by promoting generation of CD4+CD25+ T regulatory cells which have important role in immune tolerance induction and autoimmune disease prevention. MSCs are able to block pro-inflammatory and increase anti-inflammatory cytokines secretion. So such unique immunomodulatory features make MSCs ideal candidates for clinical application as immunosuppressants which can be considered for autoimmune diseases treatment. Therefore, in this short-review, we attempt to focus mainly on the existing information about MSCs in association with immunomodulatory function of them on the immune system. In addition, the possible mechanisms and the performance impact of MSCs in autoimmune diseases improvement are discussed here. However, increasing knowledge of how MSCs will influence on the immune system suppression, leading us to better use of these cells as a promising tool in the treatment of autoimmune diseases.