hassan noorbazargan

Peptic ulcer disease is a multifactorial disease that affects up to 10% of people. The use of natural product remedies has received much attention for its treatment. In this research, the healing effect of metabiotic extracted from Bifidobacterium bifidum was investigated.

45 male wistar rats were divided into 3 groups (Ctrl-, drug, and metabiotic), and stomach ulcers were induced by ethanol administration and treated by drug and metabiotic. The healing process was investigated on different days by histological analysis and qRT-PCR.

The metabiotic increased IL-8 and PDGF expression and stimulated the recruitment of polymorphonuclear cells to the wound site. It caused a faster onset of the inflammation phase followed by the proliferation phase. The metabiotic increased the expression of SOD and GPx genes and the antioxidant capacity of the wound. The increase in EGF expression led to faster re-epithelization, which was evident in the wound closure process.

Metabiotic extracted from B. bifidum is a promising candidate for the treatment of PUD. It causes a faster onset of the inflammation phase. Improving the antioxidant status of the wound, causes a faster resolution of inflammation, which leads to the acceleration of the wound-healing process.

Emphasis on substitutional medications with the elevated bacterial resistance to current antibiotics is pivotal. We evaluated the antibacterial effect of AK2 by Minimum Bactericidal Concentration (MBC) and Minimum inhibitory Concentration (MIC) and its impact on macrophage responses in 17 strains of pathogenic bacteria. The gene expression of macrophage’s cytokines was evaluated. Accordingly, the bioinformatic assessment predicted this peptide’s physicochemical characteristics, behavior, and structures. The present study aimed to assess the antibacterial effect of AK2 peptides on Macrophage responses.

Cytotoxicity level was assessed by MTT assay on the HeLa cell line. The hemolytic activity of peptides on red blood cells was evaluated. The Griess assay was performed to assess the amount of macrophage nitric oxide production. The real-time PCR method measured the iNOS, IFN-γ, and TNF-α gene expression in isolated macrophages. 

Peptide concentrations (13-60 µg/mL) were observed as the MBC and MIC value results for various bacteria. No remarkable cytotoxicity was observed at 30 and 60 µg/ml concentrations after 24h. iNOS, IFN-γ, and TNF-α gene expression were upregulated. There was also a higher secretion of nitric oxide in 48 hour-culture of the cell line with peptide. Great antibacterial activity was observed in some bacterial strains, particularly B. melitensis. 

AK2 peptides display suitable antibacterial activity with negligible toxicity for host cells. This peptide could also stimulate macrophage responses through nitric oxide production and gene expression in proinflammatory cytokines.

Torque Teno virus or transfusion-transmitted virus (TTV) is a non-enveloped virus with a single strand circular DNA genome that currently is classified in the Alphatorquevirus genus and the family of Anelloviridae. Unlike other DNA viruses, TTV has an extremely wide genomic diversity. This virus, based on previous studies, infects both healthy people, as well as those who have HCV and human papillomavirus (HPV). This study aimed to evaluate the coinfection of torque teno virus (TTV) and HPV in cervical samples from Iranian women.

In this case-control study, the fresh cervical cytobrush specimens were collected from 150 women referred to Dena laboratory in Tehran. Viral DNA was extracted from samples. The HPV-DNA was detected and genotyped. Then, nested polymerase chain reaction (Nested PCR) was performed for TTV using specific primers.

Among 50 cervical specimens without HPV, 14 were TTV positive (28%); among 50 low-risk HPV cervical specimen, 23 were TTV positive (46%), and from 50 high-risk HPV cervical specimen, 48 were TTV positive (96%). There is a significantly higher prevalence of TTV virus in low-risk and high-risk papillomavirus-infected specimens than in healthy specimens (p 0.0001). Additionally, TTV is more prevalent in samples containing high-risk papillomaviruses than in samples with low-risk papillomaviruses (P = 0.048).

The higher prevalence of TTV among people infected with papillomavirus than in non-infected people indicates that both viruses are transmitted by the same mechanism (sexual route). In addition, the prevalence of TTV in samples containing high-risk papillomavirus is significantly higher than that in samples containing low-risk papillomavirus. The presence of papillomaviruses, particularly high-risk types, may be associated with TTV proliferation, which requires further research in the future.

Klebsiella pneumoniae is one of the most important causes of nosocomial infections, especially wound infection after surgery. One of the mechanisms of antibiotic resistance in K. pneumoniae strains, especially ciprofloxacin, is the presence of efflux pump.

The aim of this study was to evaluate the anti-efflux activity of Grammosciadium platycarpum Boiss. & Hausskn. extract on the expression of oqxA efflux pump in K. pneumoniae strains which were resistant to antibiotics.

In this experimental study, clinical specimens were collected from hospitals in Tehran and the K. pneumoniae strains were isolated. Subsequently, ciprofloxacin-resistant strains containing the oqxA efflux gene were detected using PCR. Finally, the gene expression of oqxA efflux pump in the strains treated with G. platycarpum extract was investigated using Real Time PCR.

In this study, 50 K. pneumoniae strains were isolated from clinical specimens and the results of antibiotic susceptibility showed that 70% (35 strains) of isolates were resistant to ciprofloxacin and oqxA gene was observed in 43% (15 strains) of ciprofloxacin resistant K. pneumoniae strains. Moreover, Real Time PCR results showed that the expression of oqxA gene in the strains which are treated with extract, down-regulated significantly.

The results of this study showed that the G. platycarpum extract can inhibits the expression of the oqxA efflux pump in K. pneumoniae strains, and with further studies, the G. platycarpum extract can be used as a candidate for the drug design.

The treatment of bladder cancer is usually performed by Bacillus Calmette-Guerin (BCG) instillation. BCG therapy is a common therapeutic method with fewer side effects compared with chemotherapy, radiotherapy, etc. BCG can also inhibit the progression and recurrence of bladder cancer by inducing apoptosis pathways, arrest cell cycle, autophagy, and neutrophil extracellular traps (NETs) formation. However, BCG therapy cannot be effective in the situation that the metastasis occurs. NETs are induced by BCG and help suppress the growth of tumor cells, especially in the primary stages of bladder cancer. Activated neutrophils can stimulate cellular pathways, such as autophagy and NETs release in the presence of microbial pathogenesis, inflammatory agents, and tumor cells. Autophagy can also regulate NETs formation and induce Reactive Oxygen Species (ROS) and NETs production. Moreover, miRNAs are key regulators of gene expression. These small non-coding RNAs are also considered as an essential factor in controlling tumor development. The interaction between BCG and miRNAs is still unclear. However, in the present study and for the first time, we intend to discuss the role of miRNAs in BCG therapy and how NETs formation can be effective on BCG performance to treat the bladder cancer.

Staphylococcus aureus is one of the important nosocomial infection agents. Recently, S. aureus strains have become resistant to ciprofloxacin and the efflux pump is considered as its contributor. Herein, we investigated the presence, expression, and activity of efflux pump genes (norA and norB) among ciprofloxacin-resistant S. aureus isolates.

A total of 250 clinical samples were subjected to isolation of S. aureus strains. The antibiotic resistance pattern was characterized and the presence and expression level of norA and norB genes was assessed using PCR test and real-time PCR test, respectively. Finally, active efflux pumps were detected in ciprofloxacin-resistant S. aureus strains using the ethidium bromide test.

Among total clinical samples, 50 S. aureus strains were recovered. Of this 12 samples (24%) were resistant to ciprofloxacin. Moreover, norA and norB genes were found in 100 % and 83% of ciprofloxacin-resistant isolates, respectively. All ciprofloxacin-resistant isolates exhibited active efflux pumps. Real-time PCR results revealed that the isolates are more resistant to ciprofloxacin having a high level of efflux pump gene expression.

The results of this study showed that norA and norB efflux pump genes play an important role in resistance to ciprofloxacin in S. aureus strains.

Ciprofloxacin resistance in Staphylococcus aureus strains due to efflux pumps has become a significant challenge. This study was performed to evaluate the frequency and gene expression of norA and norB efflux pump genes and their role in resistance to ciprofloxacin in clinical isolates of S. aureus. In this experimental study, a total of 250 clinical samples were collected from different hospitals in Tehran and S. aureus isolates were identified. Antimicrobial susceptibility patterns were determined by disk diffusion method based on CLSI standard. The presence of norA and norB efflux pump genes in ciprofloxacin isolates were detected using PCR method. Finally, active efflux pump was evaluating using MIC of ciprofloxacin-Ethidium bromide and Real Time PCR method. Among 250 clinical samples, 50 S. aureus isolates were recovered and the results of antibiotic susceptibility tests show that 34 out of 50 S. aureus isolates (68%) were resistant to methicillin (MRSA) and from the 34 MRSA, 12 isolates (24%) were resistant to ciprofloxacin. Moreover, the norA and norB genes were found in 100 % and 83% in isolates, respectively. Real Time PCR results show that more resistant strains had increased expression in norA and norB efflux genes. The potential role played by norA and norB efflux pumps in the development of resistance to ciprofloxacin in clinical isolates of S. aureus and the detection of these genes could be important for suggestion of an effective treatment model for the S. aureus infections.

norA efflux pump is one of the mechanisms of ciprofloxacin resistance in Staphylococcus aureus strains, and finding natural herbal compounds with the ability to inhibit efflux pumps is one of the recent challenges. The aim of this study was to determine the inhibitory effect of Artemisa quttensis extract on norA efflux pump gene expression in ciprofloxacin resistant S. aureus strains.
At first, A. quttensis ethanolic extract, was prepared by ethanol solvent. Then, existence of norA efflux pump was detected in 10 ciprofloxacin resistant clinical strains of S. aureus using polymerase chain reaction (PCR) method. Moreover, anti-efflux activity of the extract was determined using ethidium bromide method. Finally, after treatment of isolates with Sub-MIC concentration of the extract, norA gene expression level, was evaluated using real-time PCR method.
The PCR results showed that all of the strains had norA efflux pump, and and ethidium bromide method revealed that the A. quttensis extract had inhibitory effect on resistant strains that had norA efflux pump. Moreover, real-time PCR results showed that norA gene expression level was down-regulated by Sub-MIC concentrations of the extract in the ciprofloxacin resistant isolates.
Considering the anti-efflux pump effect of A. quttensis extract, it seems that this extract has the potential to be used as a native plant in pharmaceutical industries.

Medicinal plants have been identified and used from prehistoric times and these plants make many chemical compounds for biological functions. Trifolium cherleri is an herbaceous species belonging to the family of the Fabaceae to Africa, Eurasia and Australia. T. cherleri is an important member of the Fabaceae family that is well-known herbal medicine in Iran. The aim of this study was to investigate the phytochemical composition, antibacterial and anti-cancer activities of T. cherleri extract.
This experimental study was performed in Islamic Azad University, from December 2016 to February 2017. At first, the phytochemical constituents of T. cherleri extract were determined using gas chromatography-mass spectrometry (GC-MS) method. Subsequently, the antibacterial activity of the extract was evaluated against some gram positive and negative pathogenic bacteria included Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC 19615, Salmonella enteritidis ATCC 13076 and Listeria monocytogenes ATCC 35152 via minimum inhibitory concentration (MIC) method. Moreover, anticancer potential of extract was examined by colorimetric MTT assay toward lung cancer (A549) cell line. Then, the evaluation of caspase 3 and 9 apoptosis gene expression was determined using Real-Time Polymerase Chain Reaction (Real-Time PCR) technique. Moreover, the Real-Time PCR was performed using relative quantitative method.
The phytochemical analyses of T. cherleri extract showed the 20 major components and the most frequent component was belonged to hexadecanoic acid, ethyl ester (20.7%) and 2-Pentadecanone, 6,10,14-trimethyl (19.9%). The extract had maximum antibacterial effects against Staphylococcus aureus and Streptococcus pyogenes. There was a dose dependent increase in the cytotoxicity effect of extract against A549 cancer cell. Moreover, the Real-Time PCR results indicated that the caspase 3 and caspase 9 gene expression was significantly up-regulated 2.57±0.27 (P The results of this study showed that the T. cherleri extract had significant anti-bacterial and anti-cancer effects and it appear that the extract has potential uses for pharmaceutical industries. Moreover, it could be considered as a promising source for novel drug compounds, but more studies are needed.

Staphylococcus aureus is a major causative agent of nosocomial infections which produces a wide range of diseases due to biofilm formation. This study aimed to investigate the phytochemical composition of Helichrysum artemisioides extract, its anti-biofilm effects on clinical isolates of ­methicillin-resistant Staphylococcus aureus, as well as, analysis of icaD gene expression.
First, the phytochemical composition of H. artemisioides extract was studied by GC/MS method. Then, the biofilm formation capacity of 50 clinical isolates of S. aureus was evaluated by Congo red agar and polymerase chain reaction (PCR). Finally, after treatment of strains with SubMIC ­concentration of the extract within 24 hours, the expression level of icaD gene was evaluated by Real Time PCR.
Findings: The phytochemical analysis of extract showed 52 compounds, including α-Pinene (12.5%) and Carvacrol (8.6%) as the major components. The evaluation of biofilm formation test shows that out of 50 strains of S. aureus, 10 strains (20%) were positive for biofilms formation. Also, after treatment of selected strains by SubMIC concentration of the extract, the gene expression quantitation of icaD was down-regulated significantly (PDiscusion & Due to the significant effects of H. artemisioides extract on biofilm formation, it seems that the extract could be used as complementary therapy to inhibit S. aureus biofilm.

Nowadays, the probiotic bacteria such as lactobacilli are known as prevention factor for various disease especially cancer. The aim of this study was to investigate the cytotoxic effect of Lactobacillus casei PTCC 1608 cell extract as probiotic bacteria on colon cancer cell line (HT29) and analysis of Bax and Bcl2 apoptosis gene expression.
In this experimental study, the cell extract of heat killed L. casei was prepared in 0.01, 0.1, 1, 10, 100 and 1000 µg/ml concentration and subsequently, the cytotoxicity of various cell extracts on HT29 and HEC293 cell lines were evaluated in 24 hours using MTT assay. Moreover, the Bax and Bcl2 apoptosis gene expression level in HT29 cell line was analyzed using Real Time PCR. The apoptotic effects of cell extract was determined using Flow-cytometry technique. Finally, the collected data were statistically analyzed using one-way anal­ysis of variance with the SPSS/18 software.
The results of MTT test show that cell extracts of L. casei is able to reduce the survival rate of HT29 cell line to 0.95±0.44, 73.45±0.21, 51.49±0.87, 39.5±0.45 and 19.7±0.55. In addition to, the Real Time PCR results indicated the expression level of Bax and Bcl2 was increased and decreased respectively, in HT29 cell line (2.76 ± 0.54 (P The results show that the cell extract of L. casei PTCC 1608 could induced the apoptosis in HT29 cell line and it had low toxicity on HEC293 cell line. Therefore, it seems that L. casei has potential uses as probiotic for pharmaceutical applications including prevention and treatment of colon cancer.

Aloysia citrodora belongs to the Verbenaceae family of plants, a well-known herbal medicine in Iran. The aim of the present study was to investigate the chemical composition, antioxidant, antibacterial, cytotoxic and apoptotic effect of A. citrodora extract against human colon cancer (HT29) cells by using real-time polymerase chain reaction and flow-cytometry methods.
This experimental study was carried out in Islamic Azad University, East Tehran Branch, from March to September of 2014. At first, the A. citrodora chemical constituents were analyzed by gas chromatography-mass spectrometry (GC-MS) technique. In addition, antioxidant assay, antibacterial and anti-cancer effect was performed using 1,1-diphenyl-2-picrylhydrazyl (DPPH), disk diffusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods, respectively. The half maximal inhibitory concentration (IC50) value was calculated. We extracted total RNA molecules by using RNX solution, after which cDNA was synthesized. Finally, the pro-apoptotic (Bax) and anti-apoptotic (Bcl2) gene expression was performed by real-time polymerase chain reaction and apoptotic effects were analyzed using Flow-cytometry method.
GC-MS analysis of Aloysia citrodora extract was shown 37 major components and the most frequent component was belonged to Spathulenol (17.57%) and Caryophyllene oxide (15.15%) The antioxidant activity of the extract was IC50= 0.6±0.03 mg/ml. The maximum and minimum antibacterial effects of extract were belonged to gram-negative and gram-positive bacteria, respectively. Cytotoxic results revealed that the A.citrodora extract have IC50= 20.1±0.78 mg/ml against colon cancer (HT29) cell line and real-time polymerase chain reaction results showed the expression level of Bax and Bcl2 was increased and decreased respectively in colon cancer cell line (3.470±0.72 (P According to the results, it seems that A. citrodora extract has potential antioxidant, antibacterial and anticancer effects and it suggested that further studies were performed for A. citrodora pharmaceutical importance.

Human papilloma virus is a DNA virus from the papillomavirus family that is most prevalent in human cervical cancers and many studies showed the E6 and E7 proteins are present in the majority of cervical cancer cases. Development of universal HPV peptide-based vaccine with more serotypes coverage has considerable value. The aim of the study was to design a multi-epitope universal vaccine for major HPV based on E6 and E7 proteins and optimization the expression of polytopic construct contains E6 and E7 genes from different genotypes of human papilloma virus as a candid vaccine. In this experimental study that was carried out in Pasteur Institute of Iran, Virology Department from October 2013 to November 2014. In order to design the polytypic construct, we predicted the most probable immunogenic epitopes of E6 and E7 from common high risk HPV16, 18, 31, 45 along with high prevalent type 6 and 11 using bioinformatics methods. The synthetic pET28a expression vector harboring E6 and E7 protein was transformed into Escherichia coli hosts and its expression was analyzed by SDS-PAGE and western blotting. Finally, in order to expression optimization of recombinant protein, cell density, induction time, growth temperature, IPTG (Isopropyl β-D-1-thiogalactopyranoside) concentration and cultures media were studied. In the present study the recombinant fusion protein was expressed successfully and the highest expression of target protein was achieved in super broth medium containing 0.1% glucose and 0.2% L-arabinose. In Super broth medium, the optimum condition for recombinant protein expression was occurred at OD600 of 0.8, 0.1mM IPTG, one hour’s incubation time at 37 °C and BL21 (A1) host. The results of this study show that the optimum expression of E6 and E7 proteins from different genotypes of human papilloma virus can be performed. Moreover, by purification of recombinant protein and evaluation of its immunogenicity in mice, it can be used as a vaccine candidate against the human papilloma virus.

Core+1 or F protein is recently identified to be encoded by hepatitis C virus (HCV) genome. Although functional properties of this protein are not still discovered, core+1 seems to play important roles in viral life cycle and pathogenesis. This study seeks to clone and express HCV core+1 gene in a eukaryotic system with the final aim of evaluating its potential roles in cell signaling pathways and resistance of HCV to therapy.

Core+1 gene corresponding to the amino acids 1-162 was PCR-amplified from the original gene of subtype b that was previously cloned in a prokaryotic vector (14). The amplicon harboring 6xHis-tag at C-terminal was subsequently subcloned in the eukaryotic pCDNA3.1+ vector. The Nhe I / BamHI restriction sites, initiation and stop codons as well as kozak sequences were designed in the primers. Constructed plasmid was verified by restriction enzyme analysis and sequencing reactions. Liver cell line of Huh7 was transfected with this plasmid using electroporation and lipofection techniques. Transfection efficiency was checked by co-transfection of pEGFPN1 plasmid and flowcytometry. Finally expression and localization of core+1 protein was evaluated by means of confocal microscopic technique.

Restriction enzyme analysis and sequencing reactions indicated the accuracy of cloning procedure. Flow cytometric analysis showed higher electroporation efficiency for transfection of Huh7 cells, in comparison with lipofection method. Finally, results of confocal microscopic microscopy using anti-His antibody confirmed the transcription and expression of core+1 protein in Huh7 cells.

Our study resulted in the eukaryotic expression of core+1 protein in Huh7 cells and provided an appropriate tool for future studies on the potential interference of core+1 with intracellular signaling pathways and cellular protein interaction.