majid sadeghizadeh

Artemia or brine shrimp is a small crustacean found in different parts of the world except the Arctic. This research was done considering the importance of molecular studies in identifying the species populations and the genetic diversity in bisexual and unisexual forms of Artemia (together and among populations).

Artemia were sampling from 9 regions (Shoor, Inche Borun, Namak, Hoze Sultan, Mighan, Maharlu, Bakhteghan, Urmia and Nogh Lakes) and 315 samples were used according phenol-chloroform method for DNA extraction. The primers were designed based on the sequence of Artemiaʹs mitochondrial ribosomal gene and PCR was performed. Enzymatic digestion of the PCR product was performed with 10 enzymes (AluI, Eco47I, HaeIII, HindIII, HinfI, MobI, MspI, RsaI, TaqI, EcoRI). After identifying genotypes and calculating haplotypes, haplotype and nucleotide diversity within a population, nucleotide diversity and divergence between populations and genetic distance between haplotypes with Reap analysis software and frequency of geographic heterogeneity of haplotypes with Chi-square test and Monte-Carlo simulation were calculated.

This study showed the presence of 25 different haplotypes, including 9 haplotypes in Urmia, 4 in Shoor and Inche Borun, 4 in Nogh, 1 in Namak and Hoze Sultan, 3 in Mighan, 1 jointly in Bakhteghan and Maharlu, and 3 in Maharlu. The lowest haplotype diversity in the samples was found in Hoze Sultan, Namak and Bakhteghan and the highest amount was seen in Maharlu. The lowest amount of nucleotide diversity in the samples belonged to Hoze Sultan, Namak and Bakhteghan and the most belonged to Urmia. In nucleotide diversity among the samples, the lowest value was observed between Hoze Sultan and Namak and the highest value was observed between Inche Borun and Shoor with Nogh. Nucleotide divergence between the samples was the lowest for the Inche Borun and shoor and the highest value for the Inche Borun and Shoor with Nogh. In the evolutionary distance between haplotypes, the highest amount belonged to Nogh and Mighan haplotypes with Inche Borun and Shoor haplotypes.

The study of population separation based on the frequency of haplotypes showed a significant statistical difference, except in the comparison of Hoze Sultan with Namak and Inche Borun with shoor (p<0.001), and at the haplotype level, it is possible to separate the Artemia population in Iran into 7 population as Hoze Sultan - Namak, Mighan, Maharlu, Bakhteghan, Nogh, Urmia and Inche Borun - Shoor was provided.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen that causes various infections and exhibits antibiotic resistance and virulence factors, requiring alternative therapies. This study aimed to evaluate the effects of nanocurcumin on gene expression of S. aureus isolates from burn wounds in Iraqi patients, focusing on inhibiting resistance and virulence genes.    From March 2023 to May 2024, burn wound samples from Iraqi patients yielded 110 S. aureus isolates. Identification was conducted by Gram staining, biochemical assays, and culture techniques. Fifty isolates were randomly selected for antibiotic susceptibility testing using the VITEK 2 Compact System. Ten isolates showing the highest resistance to multiple antibiotics were selected for molecular characterization via Multiplex polymerase chain reaction (Multiplex PCR) to detect fnbA, icaA, icaB, ftsZ, hla, pvl, femA, and mecA genes. The ten isolates were then divided into two groups: a treatment group exposed to nanocurcumin and an untreated control group. The MIC (minimum inhibitory concentration) of nanocurcumin was determined using the broth microdilution method in a 96-well plate. The 16S rRNA gene served as an internal control for evaluating the molecular effects. A two-tailed t test was used to assess the significance of gene expression differences. All 110 isolates were confirmed as S. aureus. The 50 selected isolates were resistant to cefoxitin, amoxicillin, benzylpenicillin, ampicillin/sulbactam, piperacillin/tazobactam, cloxacillin, oxacillin, and azithromycin. MecA gene was detected in all isolates. Among the ten tested, femA, icaA, hla, and ftsZ were present in 70%; pvl in 50%; icaB in 20%; and fnbA in 10%. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), showed significant downregulation of icaA, hla, pvl, femA, and mecA in treated isolates. No significant changes were seen in fnbA and ftsZ.  Nanocurcumin inhibits S. aureus virulence and resistance genes, reducing biofilm formation and toxin production, but lacks effect on fnbA and ftsZ, requiring further research.

Macrophages are considered a particularly challenging cell type to transfect. Given their importance as therapeutic targets, developing a successful transfection method for these cells is highly desirable.

The efficiency of lentiviral transfection was compared to three commercially transfection reagents (Xfect™ Transfection Reagent, FuGENE® HD, and Lipofectamine TM 3000) in RAW264.7 macrophage cells. Following optimization and production of lentiviral particles in 293T cells, RAW264.7 cells were infected with varying MOI. Transduction efficiency, cell viability, and metabolic activity were measured and compared to the transfection efficiency of the chemical methods.

None of the three chemical transfection reagents successfully transfected RAW264.7 cells. In contrast, the lentiviral method achieved transduction even at the lowest concentration of viral stock, with a green fluorescent signal observable under a fluorescence microscope. Increasing the viral stock concentration and using higher MOIs (up to 30) significantly (p≤0.0001) increased transduction efficiency.

Despite requiring more time and effort than chemical methods, lentiviral transduction exhibited superior efficiency in transfecting hard-to-transfect cells and further improvements were achieved through some modifications such as virus concentration, the use of polybrene, no viral freezing and O/N incubation with concentrated viral particles. Since other parameters, especially the use of retronectin and spinoculation, are effective on the efficiency of the virus infection process, it is suggested that they be considered in future studies and given the encouraging data of this research, this completed method can also be applied to other difficult-to-transfect cells, such as different types of stem cells or primary cells.

Mutations in the p53 gene have been linked to the initiation and progression of breast cancer, as well as resistance to chemotherapy. Therefore, the development of novel treatment approaches is essential to combat this disease.

This study aimed to evaluate the effects of dendrosomal curcumin (DNC) on the breast cancer cell line MDA-MB231.

MDA-MB231 cells were treated with 20 μM DNC, and the apoptosis rate and cell proliferation cycles were assessed using flow cytometry. Additionally, after RNA extraction and cDNA synthesis, the expression levels of Lnc-DANCR, EZH2, Noxa, bcl- 2, bax, PUMA, p21, and p53 genes were analyzed using RT-PCR. Protein expression levels of P53, P21, Bcl-2, and Bax were evaluated through western blotting.

Dendrosomal curcumin induced apoptosis in MDA-MB231 cells and caused cell cycle arrest at the SubG1 phase. Dendrosomal curcumin treatment downregulated Lnc-DANCR, EZH2, bcl-2, and p53 gene expression, while upregulating bax, Noxa, PUMA, and p21 gene expression in a time-dependent manner. Bax and P21 protein levels were significantly upregulated following DNC treatment, whereas Bcl-2 and P53 protein levels were downregulated in DNC-treated breast cancer cells.

In summary, dendrosomal nanocurcumin demonstrated potent anti-tumor effects against breast cancer cells, suggesting its potential as a therapeutic agent in breast cancer treatment.

Breast cancer is the most common cancer in women, and despite many scientific advances, it remains the leading cause of cancer-related death in women. To solve this global problem, deeper molecular studies in the field of breast cancer are needed. Nowadays, the role of piRNAs in various cancers is of great interest. In this study, we aim to identify important piRNAs involved in breast cancer. For this purpose, raw small RNA seq data related to cancerous and normal breast tissue samples were selected and extracted from the GEO database, and the Galaxy platform was used for their bioinformatic analysis. The differential expression of 372 piRNAs was obtained based on Log2 FC ≥ 2, p-value ≤ 0.05, of which 191 showed increased expression and 181 showed decreased expression. The highest increase is related to hsa-piR-33125, whose target is GATAD2A and plays a role in carcinogenesis processes such as blood vessel development, apoptosis, regulation of gene expression at the transcriptional level, etc. The largest decrease is related to hsa-piR-33073 with Log2 FC= -4.20. To find a list of important piRNAs that have a significant difference in expression in breast cancer compared to normal tissue, as well as to determine the increase or decrease of their expression in cancer tissue and to identify the target genes and investigate their role in the biological pathways involved in the development and progression of cancer. This can be the beginning of studies that will ultimately lead to advances in breast cancer research and treatment methods.

 Amyloid beta (Aβ) is the major constituent of harmful plaques in the Alzheimer’s patients. Thus, study of Aβ and understanding its related molecular and cellular mechanisms is essential for diagnosis and therapeutic interventions. This study introduces a rapid, simple, and cost-effective technique for production and purification of this peptide, utilizing the expression of Aβ gene within bacterial system.

Aβ gene was synthesized and transferred into the expression vector pET26b. After induction by Lactose and  24 hours of incubation for Aβ expression the cell sediment was analyzed for presence of recombinant peptide using SDS-PAGE and Western blot.  Then the purification of recombinant peptide was carried using nickel chloride affinity chromatography. Characterization of purified Aβ was performed by evaluating cell cytotoxicity in 25 µM and 50 µM concentrations using MTT assay on Alzheimer cell line model SH-SY5Y.

Colony PCR and sequencing results showed the correct insertion of Aβ coding fragment into the expression vector. Presence of bands with the expected size in the results of SDS PAGE and western blot had confirmed successful expression of his-tagged recombinant peptide. MTT assay results showed the purified peptide has respectively 30 and 50% cytotoxicity for 25 µM and 50 µM concentrations.

Production of amyloid beta peptide in bacterial hosts seems to be favorable. Obtaining Aβ peptide in soluble phase is an important advantage of this study. Hence according to toxicity of the purified peptide, it can be utilized for cell line treatments and further researches on Alzheimer disease.

Breast cancer (BC) is the leading cause of cancer-associated mortality in women worldwide. However, the molecular mechanism underlying the process is still unclear. In this regard, bioinformatics studies play a decisive role in facilitating the path of biological investigations and can ultimately lead to the identification of better molecular candidates for further study.

Due to the abnormal expression of many coding and non-coding genes in all types of cancers and their relationship with various mechanisms of carcinogenesis, this study aimed at evaluating the expression levels of certain coding and non-coding genes involved in BC based on bioinformatics findings and laboratory investigations.

Gene expression dataset, module extraction, functional enrichment analysis, protein-protein interaction network construction, and RT-qPCR were performed based on bioinformatics methods and laboratory investigations. Additionally, the promoter region mutations of these genes were investigated, using sequencing of extracted DNAs from formalin-fixed paraffin-embedded (FFPE) tumor tissues.

A module was selected as a candidate for further investigation. Estrogen receptor 1 (ESR1) and forkhead box A1 (FOXA1) showed the highest degrees in the PPI network with 9 and 7 links, respectively. Furthermore, the expression levels of the FOXA1 gene, RNA component of mitochondrial RNA processing endoribonuclease (RMRP), and nuclear enriched abundant transcript 1 (NEAT1) were significantly upregulated in the tumor group compared to the control group (in order, P = 0.044, P = 0.014, and P = 0.0004). The tumors of patients with positive metastasis displayed significantly higher levels of NEAT1 and RMRP expression compared to those of negative metastasis samples (P < 0.05). Moreover, the expression level of RMRP dramatically decreased in HER2-positive patients compared to negative samples (P = 0.011). Finally, no mutations were observed in the promoter sequencing of positive metastasis samples compared to normal samples.

The upregulation levels of all three examined genes may correlate with BC progression. Therefore, they could potentially be used as biomarkers for detecting BC development.

The countries’ social and economic conditions have been threatened by corona epidemic and a large number of people’s death in the world.  SARS-CoV-2 virus, a form of corona virus family, is responsible for corona disease and its spread in the present century. The study of the receptor binding region (RBD) in the spike protein is very important for scientists because the new corona virus uses its surface spike protein for binding to the ACE2 surface protein and entering its genetic material to the host cells. By this protein and its receptor binding region inhibition, the prevention of virus entrance to the cell is possible. The virus’s genes can be multiplied by cloning and its protein can be purified. The usage of antiviral peptides as the most practical methods and binding inhibitory peptides of RBD to the ACE2 receptor for SARS-CoV-2 treatment, are of great interest to scientists. In the present research, RBD cloning in PET28a expression vector, RBD protein expression and GFP/RBD fusion protein were performed in prokaryotic host. Due to this protein’s insolubility in the prokaryotic host, column refolding was performed with urea gradient with a nickel-agarose column and the synthesized protein was confirmed through western blot technique. Three nominated peptides from articles used to compare their binding to RBD using bioinformatics and their tendency to bind to each other was investigated by molecular docking. The mentioned peptides can be used in this virus infection treatment due to their binding potential to RBD, if their interaction is proven.

Type 1 diabetes (T1Ds) is an autoimmune disease in which the immune system invades and destroys insulin-producing cells. Nevertheless, at the time of diagnosis, about 30-40% of pancreatic beta cells are healthy and capable of producing insulin. Bi-specific antibodies, chimeric antigen receptor regulatory T cells (CAR-Treg cells), and labeled antibodies could be a new emerging option for the treatment or diagnosis of type I diabetic patients. The aim of the study is to choose appropriate cell surface antigens in the pancreas tissue for generating an antibody for type I diabetic patients.

In this bioinformatics study, we extracted pancreas-specific proteins from two large databases; the Human Protein Atlas (HPA) and Genotype-Tissue Expression (GTEx) Portal. Pancreatic-enriched genes were chosen and narrowed down by Protter software for the investigation of accessible extracellular domains. The immunohistochemistry (IHC) data of the protein atlas database were used to evaluate the protein expression of selected antigens. We explored the function of candidate antigens by using the GeneCards database to evaluate the potential dysfunction or activation/hyperactivation of antigens after antibody binding.

The results showed 429 genes are highly expressed in the pancreas tissue. Also, eighteen genes encoded plasma membrane proteins that have high expression in the microarray (GEO) dataset. Our results introduced four structural proteins, including NPHS1, KIRREL2, GP2, and CUZD1, among all seventeen candidate proteins.

The presented antigens can potentially be used to produce specific pancreatic antibodies that guide CARTreg, bi-specific, or labeling molecules to the pancreas for treatment, detection, or other molecular targeted therapy scopes for type I diabetes.

Tamoxifen (TAM) is routinely used for the treatment of estrogen-positive breast carcinoma. Approximately 40% of patients with metastatic breast cancer will develop resistance to TAM. TAM therapeutic failure has been a major challenge in the treatment of TAM-resistant breast cancer cells. Therefore, finding a way to eliminate TAM resistance is very valuable. Curcumin is a polyphenol extracted from the rhizomes of Curcuma longa and has extensive biological and pharmacological effects on many cancers. The purpose of this study was to look into the effects of dendrosomal nano-curcumin (DNC) on cell growth and apoptosis, as well as the effects of DNC on the expression levels of long non-coding RNA CCAT2 in TAM-resistant MCF-7 cells (TAM-R). TAM-R cells were created, and CCAT2 expression was evaluated in TAM-R compared to TAM-sensitive MCF-7 cells (TAM-S). Forty eighth hours after TAM-R treatment with 20 μM of DNC, Q-RT-PCR, and flow cytometry cell cycle and Annexin V-PI assays were performed. P-value < 0.05 was defined as statistical significance. CCAT2 was significantly upregulated in TAM-R compared to TAM-S. DNC administration downregulated CCAT2 expression, and markedly suppressed cell cycle and induced apoptosis in TAM-R. Furthermore, DNC decreased the anti-apoptosis gene (BCL-2) and increased the apoptotic gene (BAX) expression levels respectively in TAM-R. DNC promoted cell cycle arrest and apoptosis, eventually by CCAT2 downregulation in TAM-R. However, the probable mechanisms of how DNC affects CCAT2 expression levels are unknown and need future studies.

Targeting the lytic cycle of the Epstein-Barr virus (EBV) has been considered a new treatment strategy for malignancies caused by this virus.  This study aimed to investigate the effect of Dendrosomal NanoCurcumin (DNC) to prevent cell transformation and inhibit the expression of viral lytic gene expression in the generation of lymphoblastoid cell line (LCL). Cell viability of LCLs and PBMCs was performed by MTT assay, and flow cytometry (Annexin/PI) was used for evaluation of apoptosis. CD markers on the surface of generated LCL (CD19) cells were examined for cell validation. The effect of DNC on transformation was evaluated by examining cell morphology and determining the expression level of lytic genes BZLF1, Zta, BHRF1, and BRLF1 of EBV using Real-time PCR. Student’s t-test was used for statistical analysis. The MTT assay showed that DNC can inhibit the proliferation of LCL in a dose-dependent manner. The 50% cytotoxic concentration (CC50) of DNC and curcumin for LCL was determined 38.8 µg/ml and 75 µg/ml, respectively after 72 hr. Also, Real-time PCR data analysis showed that DNC in 30 µg/ml concentration significantly inhibited cell transformation in the LCL and significantly reduced viral lytic genes such as BZLF1, Zta, BHRF1, and BRLF1expression compared to control. Overall, these findings show that DNC reduces the expression of the viral lytic cycle genes and also the induction of cell apoptosis and finally prevents the generation of LCL.

Chemotherapy drugs used to treat lung cancer are associated with drug resistance and severe side effects. There have been rising demands for new therapeutic candidates and novel approaches, including combination therapy. Here, we aimed to investigate the combinatorial effect of a dendrosomal formulation of curcumin (DNC) and daunorubicin (DNR) on the A549 lung cancer cell line.

We performed cytotoxicity, apoptosis, cell migration, colony-formation capacity, and gene expression analysis to interpret the mechanism of action for a combination of DNC and DNR on A549 cells.

Our results revealed that the combination of DNC and DNR could synergistically inhibit the A549 cells’ growth. This synergistic cytotoxicity was further approved by flow cytometry, migration assessment, colony-forming capacity and gene expression analysis. DNR combination with DNC resulted in increased apoptosis to necrosis ratio compared to DNR alone. In addition, the migration and colony-forming capacity were at the minimal range when DNC was combined with DNR. Combined treatment decreased the expression level of MDR-1, hTERT and Bcl-2 genes significantly. In addition, the ratio of Bax/Bcl2 gene expression significantly increased. Our analysis by free curcumin, dendrosomes and DNC also showed that dendrosomes do not have any significant cytotoxic effect on the A549 cells, suggesting that this carrier has a high potential for enhancing the curcumin’s biological effects.

Our observations suggest that the DNC formulation of curcumin synergistically enhances the antineoplastic effect of DNR on the A549 cell line through the modulation of apoptosis/necrosis ratio, as well as Bax/Bcl2 ratio, MDR-1 and hTERT gene expression.

The crucial and vital player in tumor recurrence is the tumor-initiating cells (TICs). OCT4 is a widely appreciated non-cell surface for TICs, dedicating detrimental properties to these cells, including self-renewal, epithelial-mesenchymal capacity, and drug resistance. OCT4 and its partners Sox2 and Nanog are up-regulated in stem cells; on the other hand, normal stem cells are more resistant to various herbal remedies like curcumin. Based on these facts, the main objective of the present study was to investigate the alteration of the mentioned genes expression after curcumin treatment in breast cancer cell, human bone marrow mesenchymal stem cells (hBM-MSCs), and non-tumor fibroblast cells (HSFPI3).

MTT assay and AnnexinV/PI were performed to calculate the effective concentration of curcumin. To assess the expression level of OCT4 and Nanog, real-time PCR was performed to quantify the alteration of the mRNA expression of the mentioned genes after treatment in MDA-MB231, hBM-MSCs, and HSFPI3.

Curcumin could not induce significant apoptosis in hBM-MSCs and HSFPI3 even after 24 and 36 hours after treatment in a toxic concentration for cancer cells. After 36-hour treatment with DNC, the mRNA expression of Oct4-B1 in both normal cells enhanced significantly compared to untreated samples. Furthermore, in HSFPI3 cells, the Nanog mRNA expression increased after this treatment.  The expression of both genes decreased in the MDA-MB231 after treatment with DNC.

Non-tumor cells are more resistant to the curcumin treatment compared to cancer cells. The reason is at least partially due to the different expression pattern results in these cells after treatment with this reagent. Pluripotent markers, including Oct4-B1 and Nanog are proposed to play a vital role in these non-tumor cells resistant to curcumin.

 The Islets of Langerhans include Alpha, Beta, Delta, and Epsilon cells whose secret hormones play important roles in glucose metabolism as well as some physiological processes in our own body.

In this study, we selected a microarray row data of transplanted pancreatic Islets from Gene Expression Omnibus (GEO) database. The row data of 10 individual samples was analyzed with R programing software. Top expressed genes in human pancreatic islets were chosen and the gene stable IDs were returned to gene names and descriptions by BioMart tools. The selected genes were categorized into biological processes by Protein Analysis Through Evolutionary Relationships (PANTHER) online database. Also, sub-cellular localization of their proteins was investigated by the protein atlas database.

The results showed that the quality controls of all 10 individual samples of microarray chips such as signal intensity and uniformity of the images were passed. From 336 genes, 284 proteins were classified by PANTHER online database. They are categorized into “Translational proteins”, “Metabolism enzymes”, and “Protein modifying enzymes” biological processes, respectively.

In this study, we presented 500 top-ranked expressed genes in human pancreatic islets. We also represented calcification and sub-cellular localization of these high expressed genes in separate supplementary files. Research data can be used for pancreatic research as well as potential drug design for type I diabetes or pancreatic cancers.

During research on lymphoma and its malignancies, scientists have traced the role of the angiogenesis index in patient survival. Epstein-Barr virus (EBV) is a human tumor-causing virus that targets B lymphocytes and causes persistent infection. This virus is also associated with malignancies, such as Burkitt's lymphoma. Using herbal medicines to treat cancer and angiogenesis has been considered, due to the side effects of chemical drugs. This experimentation aimed to research the antiviral impact of nano-curcumin on the Daudi cell line (which belongs to Burkitt's lymphoma) and to evaluate the expression of the vascular endothelial growth factor (VEGF) gene.

The cytotoxicity of nano curcumin, curcumin, and dendrosome on Daudi cells and normal human lymphocytes was evaluated using an MTT assay. Cellular apoptosis was assessed by Annexin / PI flow cytometry. The VEGF angiogenesis gene expression was performed by real-time PCR.

The 50% cytotoxic concentration (CC50) was determined 30 μg/ml for dendrosomal nano-curcumin, 50 μg/ml for curcumin, and 987 μg/ml for dendrosome in the Daudi cell line.Dendrosomal nano curcumin (DNC) caused time and dose-dependent death in Daudi cancer cells compared to curcumin. Dendrosome did not show toxicity on control cells. The results of Flow cytometry are constant with the results of the MTT test. The data obtained from the real-time PCR showed a significant decrease in the expression of the VEGF gene (P <0.01).

The dendrosomal nano curcumin is involved in angiogenesis by reducing the expression of the VEGF gene, and can be a good candidate as a supplement drug in the chemotherapy treatment of Burkitt's lymphoma.

Tamoxifen (TAM) is routinely used for the treatment of estrogen-positive breast carcinoma. Approximately 40% of patients with metastatic breast cancer will develop resistance to TAM. TAM therapeutic failure has been a major challenge in the treatment of TAM-resistant breast cancer cells. Therefore, finding a way to eliminate TAM resistance is very valuable. Curcumin is a polyphenol extracted from the rhizomes of Curcuma longa and has extensive biological and pharmacological effects on many cancers. The purpose of this study was to look into the effects of dendrosomal nano-curcumin (DNC) on cell growth and apoptosis, as well as the effects of DNC on the expression levels of long non-coding RNA CCAT2 in TAM-resistant MCF-7 cells (TAM-R). TAM-R cells were created, and CCAT2 expression was evaluated in TAM-R compared to TAM-sensitive MCF-7 cells (TAM-S). Forty eighth hours after TAM-R treatment with 20 μM of DNC, Q-RT-PCR, and flow cytometry cell cycle and Annexin V-PI assays were performed. P-value < 0.05 was defined as statistical significance. CCAT2 was significantly upregulated in TAM-R compared to TAM-S. DNC administration downregulated CCAT2 expression, and markedly suppressed cell cycle and induced apoptosis in TAM-R. Furthermore, DNC decreased the anti-apoptosis gene (BCL-2) and increased the apoptotic gene (BAX) expression levels respectively in TAM-R. DNC promoted cell cycle arrest and apoptosis, eventually by CCAT2 downregulation in TAM-R. However, the probable mechanisms of how DNC affects CCAT2 expression levels are unknown and need future studies.

Megakaryopoiesis is characterized by progressive polyploidization and the expression of megakaryocytic markers. Numerous transcription factors and physiological signaling pathways regulate this phenomenon. Megakaryocyte differentiation induction in the K562 cell line and hematopoietic stem cells via nanocurcumin drug has been identified in our previous study. K562 cells are typical Chronic Myelogenous Leukemia (CML) cells that are resistant to apoptosis and express the bcr-abl fusion gene. These cells have the potential to differentiate into erythrocytes and megakaryocytes. Curcumin is well known as a component with strong potential to alter NFκB activity in various cells. NFκB pathway regulates various genes such as apoptotic and immune response genes. The aim of the current study is to evaluate the possible role of nanocurcumin in NFκB pathway regulation during the megakaryopoiesis process in the K562 cell line.

Megakaryocyte markers expression and phenotype alteration of nanocurcumin-treated K562 cells have been detected by flow cytometry and microscopy imaging. The nuclear level of the RelA (p65) subunit of NFκB was determined by western blot test in K562 cells during megakaryopoiesis induction via nanocurcumin treatment at different times. The expression of NFκB target genes including c-MYC, BAX, and NQO1 was also analyzed in nanocurcumin-treated K562 cells by quantitative RT-PCR assay at different times.

It was demonstrated that nanocurcumin leads to an increase in NFκB activity transiently during megakaryocyte differentiation, which is followed by a change in the expression of c-MYC, BAX, and NQO1 target genes.

The NFκB pathway can be considered a new pathway for inducing megakaryocyte differentiation by nanocurcumin for the purposes of in vitro and in vivo megakaryopoiesis experiments.

Cervical cancer is the fourth most common cancer among women. In recent years, attention has increased to natural products such as curcumin with anti-cancer potential as a therapeutic supplement. However, due to its poor solubility, its clinical use is limited. In this regard, in this study, to improve clinical parameters, the effects of nanocurcumin on the angiogenesis inhibition of cervical cancer were investigated and compared with free curcumin.

MTT method was used to evaluate the proliferation of the HeLa cell line with free curcumin and nanocurcumin at different doses and time intervals and the rate of apoptosis was assessed by flow cytometry. Then, the expression of the vascular endothelial growth factor (VEGF-A) gene in HeLa cells was measured by Real-Time PCR and Western blotting, respectively.

According to IC50 for 48 hours in the HeLa cell line, which was 15 μM/ml and 50 μM/ml for nanocurcumin and free curcumin, respectively, the nanocurcumin showed a greater lethal effect. VEGF-A gene expression (p <0.0001) and protein level (p <0.01) were significantly lower following nano-curcumin treatment than free curcumin.

Nanocarrier increased the solubility and further inhibited the proliferation of cervical cancer HeLa cells and was three times more effective than curcumin in inhibiting angiogenesis at the same concentration. Therefore, nanocurcumin can be a good option for drug supplementation along with routine cervical cancer treatment.

Acute myeloid leukemia (AML) is a complex disease characterized by clonal expansion of undifferentiated myeloid precursors, resulting in impaired hematopoiesis and bone marrow failure. Different genetic and environmental factors are believed to be involved in the pathogenesis of AML. Notch signaling with a tumor suppressing plays a role in myeloproliferative disorder and is a negative regulator of myeloid progenitor commitment. Crosstalk between Notch signaling and CXCR4 axis is a matter of debate in AML. In the current case-control study, we evaluated the expression level of CXCR4, JAG1, and MIB1, which are all involved or related to Notch signaling in adult AML patients. Blood samples were obtained from 25 AML and 17 healthy individuals and the expression level of the selected genes was evaluated via the real-time polymerase chain reaction. Our results revealed the increased expression of JAG1, but decreased expression of CXCR4 in AML patients in Iranian population of AML patients. Moreover, some gender-associated effects on the expression of JAG1 and CXCR4 were detected, which may be related to sex hormones. The expression level of MIB1 did not change significantly. The correlation analysis showed no correlations between the age of the patients and the expression levels of the genes. Herein, for the first time, we suggested some new evidence regarding the complex role of Notch signaling-related genes (CXCR4 and JAG1) in the pathogenesis of AML in Iranian patients.

Silver carp (Hypophthalmichthys molitrix) as the major species in total production of freshwater fish, has a special place in the aquaculture industry of Iran. The common method for breeding of this species in our country is semi-natural which provides the brooders opportunities for mate selection that is one of the most striking features in this method. Given the importance of MHC gene complex to sexual behaviors and its historical role in the process of evolution of living organisms especially in terms of safety issues, various patterns of mentioned loci between two generations of Silver carp were studied. 33 males and female silver carp were reproduced in two independent groups by semi-natural method after that larvae in each group were sampled. Using data in NCBI and designing the appropriate primer, MHC-DAB loci of the parents and their F1 were studied by SSCP method. Analysis of six different genotypes between the samples indicated an increased level of observed heterozygosity (1) compare to the expected value (0.660 -0.709). Deviation from Hardy-Weinberg equilibrium was observed in most groups and the main share of genetic diversity assigned to individual differences. The pattern of genetic diversity distributed in larvae and high variation of MHC-DAB loci meanwhile retaining number of the alleles, represents a positive selection.

 Curcumin, a compound derived from the root of the Curcuma longa, has been confirmed as an anticancer, chemoprotective, and gene/protein regulatory agent. Nanoformulation of curcumin has been developed to increase its targeting efficiency, solubility, controlled release, and physical and chemical stability.

 This study investigated the effect of new nano-type curcumin, oleic acid-derived dendrosome (OA400 nanoparticles), on the expression of E6 and E7 human papillomavirus oncogenes and P53 and Rb factors in the HeLa cell line. After preparing nano-curcumin by mixing OA400 nano-carrier and curcumin, its effect was considered on the human cervical cancer cell line (HeLa cell line RRID: CVCL_003) and normal fibroblast cells.

 MTT assay and flow cytometry were used to evaluate cell viability and apoptosis. Furthermore, real-time RT-PCR and western blot analyses assessed RNA and protein expression of E6, E7, P53, and Rb. Statistical analyses were performed by GraphPad Prism 7 software.

 The nanoformulation of curcumin could reduce the expression of E6 and E7 oncogenes and increase P53 and Rb tumor suppressors in HeLa cancerous cells at 15 μM concentration; however, it had no significant effect on the viability of normal fibroblast cells. On the other hand, curcumin altered the expression of these genes at a 50-μM concentration. Gene and protein expression analysis indicated the up-regulation of P53 and Rb factors and the down-regulation of E6 and E7 under the influence of nano-curcumin treatment more than curcumin.

 These data indicate the potential of curcumin-loaded OA400 nanoparticles to be considered as a treatment option in cervical cancer investigations.

NIR Laser application in bacteria is often focused on mortality and antibiotic efficacy. The literature records on this point are absolutely diverse from mortality in different degrees to immortality and even viability enhancement. The aim of this study is to investigate 808 nm laser effects on E.coli-DH5α viability and Growth with CFU, MTT and FCM assays. To obtain the purpose, bacteria in LB media put on with 808nm laser on 100 and 200 J/cm2 dosages and were investigated and compared by CFU, MTT and FCM assay. CFU assay results after 24 hours incubation were not significantly different between laser treatments and control. (P=0.06). In contrast, MTT assay results after 1 hours from laser treatment indicated significant deleterious effects in 200 J/cm2 laser treatment compared with control(P=0.006). On the other hand, FCM assay results of laser treatments with using of PI and Triton X100 not only approved MTT assay results but also revealed some dose dependent changes on bacteria ranging from increase membrane permeability to lethal damages. As a conclusion of the results in these method assays, we can state that these different laser doses produce diverse effects on viability and growth in E.coli-DH5α. Consequently the laser treatments could be planned for antibiotic purposes or enhancing gene transformation process.

Colorectal cancer (CRC) is the second and third most common cancer in men and women respectively, and the fourth cause of cancer death of individuals. Mutations in specific genes can lead to colorectal cancer. UCA1 is one of the oncogenic genes that have been shown to stimulate cell proliferation. mTOR1 is another gene that leads to the growth of cancer cells through anabolic processes and autophagy inhibition.

In this study, we evaluate the expression of these two genes in different phases of CRC, that helps the early detection of colorectal cancer which can increase the survival rate.

First, we collected 25 colorectal cancer tumor tissues and 25 adjacent normal tissues as a control group. Then, RNA was extracted from tissue samples and cDNA synthesized. The UCA1 and mTOR1 expression was evaluated in CRC tissues compared to adjacent normal tissues by Real Time PCR.

Our results showed that the UCA1 and mTOR1 expression in the tumor tissues was significantly higher than in the adjacent normal tissues (P < 0.05). There was also a significant difference in Lynph inv and Vescu inv with mTOR1 expression (P < 0.05).

Our results showed that UCA1/mTOR1 may be important genes involved in colorectal cancer. mTOR1 was also identified as one of the possible genes in metastasis of colorectal cancer. Thus, UCA1 and mTOR1 can probably be considered as biomarkers in CRC therapy and diagnosis.

Gene expression ‎regulators and mutations are a kind of biomarker and new ‎therapeutic target for human diseases like cancer. LncRNAs are involved in Several regulatory processes. RNA components of ‎mitochondrial RNA processing ‎endoribonuclease is a lncRNA that its ‎regulatory position has been identified ‎in some cancers. Regarding the importance of the RMRP gene, we are investigating changes in the expression of this gene that are caused by promoter region mutations in breast cancer patients. In this study, LncRNA RMRP gene expression was evaluated on 25 tumor samples and 25 control samples by semi-quantitative RT PCR technique, and confirmed by Real time PCR technique for a number of samples. Also, to detect the mutation in the promoter region of the LncRNA RMRP gene, DNA extraction and sequencing were performed. The expression of the RMRP gene in tumor samples showed a significant increase relative to control samples. Expression of RMRP gene was shown a significant decrease in positive HER2 samples compared to negative HER2 specimens and a significant increase in positive metastatic tumor samples compared to negative metastatic samples . Also, sequencing of the promoter region revealed differences in the region's sequence in tumor and normal samples.Based on sequencing results, there was no mutation in the RMRP promoter sequence of the tumor . The RMRP gene can be suggested as a tumor and metastatic marker for the detection of breast cancer. But more clinical and molecular research is still needed to prove it more accurately.

Human papillomaviruses (HPVs) especially types 16 and 18 are known as the major causes of cervical cancer. Among HPV proteins, L1 and L2 capsid proteins, and also E7 oncoprotein are proposed as target antigens for vaccine design. In the recent years, the recombinant multiepitope polypeptides have attracted a special interest. The goal of this study is the design of L1-L2-E7 fusion construct and evaluation of its expression in bacterial expression system. 

In this study, the immunogenic and conserved epitopes of HPV16/18 L1, L2 and E7 proteins were selected using different bioinformatics analyses. After synthesis of the designed L1-L2-E7 fusion sequence in pUC57 cloning vector, its subcloning was performed in pET24a (+) prokaryotic expression vector using EcoRI/ HindIII restriction enzymes. The expression of the recombinant multiepitope polypeptide was done in E.coli Rosetta strain using IPTG inducer, and confirmed by SDS-PAGE and western blotting using anti-His-tag antibody. The expression was optimized under different conditions such as optical density (OD in wavelength of 600 nm), temperature and time after induction, and IPTG concentration.   

The recombinant pET-L1-L2-E7 vector was confirmed by the presence of a clear band (~525 bp) related to the L1-L2-E7 gene on agarose gel after enzyme digestion. The expression of L1-L2-E7 polypeptide in bacteria showed the presence of a clear band (~20 kDa) in SDS-PAGE and western blotting. The best conditions for expression of the recombinant polypeptide were at temperature of 37◦C, optical density of 0.7-0.8, IPTG concentration of 1mM, and time of 16 h after induction.  

The successful expression of the L1-L2-E7 multiepitope polypeptide was performed in bacterial system. In the next step, the recombinant polypeptide will be purified to use as a vaccine candidate.