سید یونس حسینی

 Determination of HPV genotypes in cervical lesions is essential in screening programs and the impact of current HPV vaccines. Therefore, we investigated the frequency of various HPV genotypes among different grades of cervical lesions.

 In total, 101 biopsy samples of different grades of cervical lesions were collected. Extracted DNA was subjected to PCR amplification of beta-globin to assess sample integrity. Duplex Taq-Man Real-Time PCR was performed to identify HPV types 16 and 18. Nested PCR following sequencing was done on those samples with HPV16/18 negative results.

The mean age of participants was 43±1. Of 101 samples, 91 (90%) were infected with HPVs. In the LSIL group, HPV genotypes 16, 18, and 51; in the HSIL group, genotypes 6, 16, 18, 31, and 53; in the SCC group, genotypes 6, 16, 18, and 3; and in the AD group, HPV genotypes 16 and 18 were detected. Also, 49 (61%) samples had co-infection with HPV16 and HPV18.

 Our results showed that HPV 16 and 18 were the most prevalent genotypes among different grades of lesions. The presence of low-risk HPV types, including genotypes 6 and 53, might need more attention in HPV screening and prevention programs.

Reservoirs operation problems have various and diverse objectives that rarely lead to a global optimized answer, and usually there are a set of optimal answers (Pareto Front). Solving these problems in the past has been possible only with the use of simpler methods, including the use of weight coefficients for various purposes and the conversion of them into a just one objective function. But in recent years, with the development of multi-objective methods, there is a good way to solve them. In this regard, the purpose of the present study was to investigate the efficiency of multi-objective optimization algorithms MOPSO, SPEA-II and NSGA-II in the reservoir operation problem using LDR, N-LDR operation models to generate the optimal Pareto front in order to sustainable development. In order to obtain the mentioned purpose, First, the NSGA-II, MOPSO, and SPEA-II optimization methods were evaluated using some benchmark problems such as standard ZDT family functions with identical conditions (equal population number and number of runs). After evaluating multi-objective optimization methods using standard ZDT family functions, NLDR, LDR and SOP operation models were coded in MATLAB programming environment and linked with multi-objective optimization methods. For each of the two optimization-simulation models (LDR and NLDR), the two objective functions were defined as follows: the first objective function was to minimize the sum of the MSI and the second objective function was the reliability maximization that the maximum of which is usually obtained under the SOP operation conditions. It turns out the two aforementioned objective functions are have a reverse relationship with each other and by increasing one of them, the other one will decrease. Reservoir operation models for a 37-year period from 1356 to 1392 were executed to find the coefficients in both linear and nonlinear models in the same conditions for each multi-objective optimization algorithm with 5000 itrations. The optimal Pareto front was obtained. By addressing obtained results, It was observed the SPEA-II optimization method in NLDR operation model, has reached to the best optimal Pareto front among others. So from its three answers were selected (by using various criteria) as sample and then sustainability indices such as reliability, resilience, vulnerability and MSI were computed for those three answers. By comparing the values of objective functions and other sustainable development criteria in SOP conditions and the NLDR-C response, it can be seen that using the multi-objective optimization model, we can by less shortages (less MSI 21, 79 compared to 32, 29), reach to approximately maximum reliability (reliability in SOP conditions). However in NLDR-A solution, the least shortage (MSI equal to 13.02) can be seen compare to other sample solutions and SOP policy. Due to research findings, it was observed the SPEA-II optimization method in all of the standard ZDT family functions has reached to a more optimal Pareto front than other optimization methods. Also this method compared to other optimization methods, has a more complete front of Pareto's solutions, which indicates the efficiency of this optimization method in multi-objective optimization problems. In the case of LDR and NLDR models with the same number of iterations, also It was observed that the NLDR model has shown better results, which can be due to the higher degree of freedom of nonlinear relationship in determining the operation policy of the reservoir releasing. In this research, a series of optimal (Pareto optimal) solutions was presented to meet the stated objectives.

High morbidity and limited therapies of hepatic fibro genesis are important factor for better understanding the molecular mechanisms of the disease. Advances in the understanding of the molecular behavior of hepatic stellate cells (HSC) allow the progress of a field dedicated to anti-fibrotic therapy. Melanoma differentiation associated gene-7 (IL-24/mda-7) as a gene induced during terminal differentiation in human melanoma cells, but the inflammatory response of cells to IL-24/mda-7 is not entirely cleared.
Materias and LX-2 cells (a human hepatic stellate cell) were treated by leptin (positive control), media (control negative), or were transfected by empty plasmid and pcDNA3.1/mda-7. The inflammatory state was evaluated through measuring the mRNA expression level of inflammatory molecule, IL-1β. The role of IL-24/mda-7 modulation on inflammatory response was assayed using SOCS1 and SOCS3 gene expressions.
The expression levels of IL-1β, SOCS1 and SOCS3 were compared in LX-2 cell line groups. The expression of the IL-1β in the transfected cells was higher than the control cell, but it was not significant. The results indicated that the expressions of SOCS1 and SOCS3 were up-regulated following pcDNA 3.1/mda-7 transfection into LX-2 cells compared to control plasmids (p=0.0179, p=0.0428).
The endogenous IL-24/mda-7 exhibited a significant modulatory effect on stellate cells. Therefore, IL-24/mda-7 and relevant signaling pathways could be employed as a target for fibrosis treatment.

Melanoma differentiation association gene-7 (MDA-7)/IL24 is a tumor suppressor gene. The iNGR peptide sequence, with excellent tropism to surface integrins has been employed for targeting of therapeutic molecules toward tumor cells. The purpose of our study was to construct a plasmid expressing modified MDA-7 fused with an iNGR peptide for better targeting to tumor cells.
At first, we amplified the MDA-7 sequence by PCR, while the reverse primer contained the iNGR peptide sequence to add it to the end of a new MDA-7 gene. The resultant MDA7-iNGR and MDA-7 were cloned into a pCDNA3.1 eukaryotic expression vector. The accuracy of cloning methods, integrity of the plasmids, and sequence were sequentially evaluated by digestions, colony-PCR, and sequencing. The expressions of the plasmid constructs were assayed by ELISA following their transfection into Ad-293 cells. Next, the plasmids were transfected into Hep-G2 cells and their mRNA were converted to cDNA. We assessed the gene expression levels of Gadd153 and Bax. As the final step, apoptosis induction of Hep-G2 cells following transfection was evaluated by the help of PI/Annexin V staining according to flow cytometry.
The results showed the integrity of construct backbone in addition to reading frame of the MDA-7-iNGR sequence. A suitable expression/secretion of modified the MDA-7.iNGR protein was detected by ELISA assay of the culture supernatant when compared to the control construct that expressed unmodified MDA-7. The viabilty test demonstrated no benefit for this kind of modification of the MDA-7 protein. Real-time PCR and flow cytometry analyses revealed that the addition of iNGR to MDA-7 caused a decrease in its apoptotic effect on hepatic tumor cells compared to the normal protein. The modified protein had significant apoptosis induction compared to the negative control group (PAlthough the new pCDNA/MDA-7.iNGR plasmid expressed iNGR-fused MDA-7 protein efficiently, it could not improve the natural apoptosis property of normal MDA-7.

Despite the cost and millions loss of life due to industrial accidents, often are preventable through risk assessment methods and control measures.
To assess the safety of gas storage tanks in Kermanshah oil refinery by Energy Trace and Barrier Analysis (ETBA).
This case-descriptive study was conducted in gas storage tanks of Kermanshah oil refinery. Energy checklist was used for identification of energy types. Energy flows were tracked and then, management and administrative procedures, and personal protective equipment were considered as safeguard. Exposed and vulnerable targets are also specified. Preliminary levels of risks were determined by combination of severity and likelihood. After suggestion of corrective action for unacceptable risks, risk assessment took place again. Identified risks were expressed using descriptive statistics such as frequency and percentage.
Findings: Overall, 74 risks and 121 dangerous energies were identified. Of these, 25 risks were unacceptable, 46 were low risk, and 3 were acceptable risk with revised. Frequency of risks related to electric power was 20, and followed by risk of displacement- pressure-volume, potential and chemical energies, with frequencies of 13, 12 and 9, respectively.
Given the environmental and protection conditions of the tanks, in addition, the high percentage of some of the damaging risks in this industry, use of appropriate control measures to prevention the event of future disasters will be inevitable.

Background & aim: One of the strategies to improve the therapeutic gene is targeting gene therapy. A method which can be considered, is adding code sequences peptide or protein with high tendency to target cells and secreting the therapeutic gene encodes a protein. However, evaluating the effectiveness of such changes in the targeted cell binding protein gene product with the usual therapeutic methods produced in prokaryotic system is directly impossible. The purpose of this study was to evaluate the design methods of cell adhesion proteins based on ELISA usable in-vitro in gene therapy.
In order to target the therapeutic gene Mda-7 by using genetic engineering, peptide coding sequence RGD4C with the tendency to cancerous cell surface integrin were inserted shortly after the artificial signal peptide sequence and the N-terminal coding region of the protein. Then, the modified and unmodified cDNA eukaryotic expression vector pCDNA3.1 were matched. Vectors were transfected in HEK-293 cell line. Then Mda-7 secreted expression levels were measured in cell culture by ELISA. After adjusting the protein concentration of Mda-7 and RGD.Mda-7, in cells transfected media, they were used as a source of protein. Reduce the concentration of these genes was assessed two hours after exposure to the integrin cell lines with HepG2, M21 and lacking integrin Saos-2 were also determined by ELISA. The present study was conducted three times independently. Data were analyzed using t-test.
Statistical analysis of the results suggested that the gene product of the gene product RGD.Mda-7 and Mda-7 to connect to HepG2 cells and M21 were more likely to have integrin. While binding to the cell lines of Saos-2, no significant difference were observed.
It seems the present ELISA based method was a suitabl e strategy for cell attachment assay in gene therapy research.

Occupational consequences and their effects on health and healthcare workers have been examined in many studies. Occupational burnout is affected by quality of life and it can be produced by long term exposures to occupational stress. The aim of this study was to determine the association between occupational burnout and quality of life among healthcare workers. This study was conducted among 80 subjects of healthcare workers of Kamyaran city in three occupational groups. Data on quality of life and occupational burn-out was collected using SF-12 and standard Geldard (GBI)questionnaires, respectively. In general, 55, 18.75 and 26.25 percent of personnel were employed in health, Nursing-Obstetric and Administrative-Services groups, respectively. Good, moderate and poor quality of life was seen in 33.75, 60 and 6.25 percent of employees, respectively. While only 22.5 percent of workers were not exposed to occupational burnout, 77.5 percent of them suffered from mild condition of this disorder. The Correlation between occupational burn-out as well quality of life with demographic characteristics was not significant. A reverse and statistical significant correlation was found between quality of life and occupational burnout (P˂0.001). Considering the reverse and significant association between occupational burn-out and quality of life, this disorder and reduction of productivity can be prevented by improving the quality of life of employees in different ways.

Chlorine is an irritant gas that has many applications including in chemical and paper industries. Although this substance is an upper and lower respiratory tract irritant، acute and chronic respiratory effects of occupational exposure to this substance have not been thoroughly investigated. Therefore، the aim of the present study was to assess the respiratory effects of occupational exposure to chlorine gas in a group of workers in the petrochemical industry. This was a cross sectional study in which a group of 54 workers with cur-rent exposure to chlorine gas (exposed subjects) and 38 healthy male office workers without history of exposure to chlorine gas (referent subjects) were investigated. Standard respiratory questionnaires were administrated to them. Additionally، the parameters of pulmonary function were measured both prior to shift and at the end of shift. The prevalence of respiratory symptoms in the exposed group was significantly higher than that of referent subjects (P<0. 05). Pulmonary function tests (before shift) showed that chlorine gas causes a significant reduction in the average of some pulmonary parameters. Spirometry (after shift) also showed that the average levels of all parameters decreased at the end of the work shift compared to the corresponding values of before exposure، although these changes were not significant. Exposure to chlorine gas is associated with irreversible chronic changes in the parameters of lung function and prevalence of respiratory symptoms.

Backgrounds and Semi automated processes of Iranian tire manufacturing factories lead to significant occupational exposures to benzene and toluene compounds. This study was conducted to evaluate the magnitude and risk of workers exposures. Personal monitoring of 100 workers (10 groups) was performed using NIOSH method No.1501 in two tire manufacturing factories in Tehran city. Workplace atmospheric conditions were measured on sampling days. Semi quantitative risk assessment (COSHH) was carried out using questionnaire and respective matrix. Data were analyzed using t-test, correlation coefficient, linear regression and one-way ANOVA. Quantitative values were reported as mean±standard error. Age, work experience and atmospheric parameters had no significant statistical difference in two factories. Occupational exposures to benzene and toluene were 1.09±0.18, 1.88±0.19 and 2.07±0.30, 3.20±0.39 ppm in factories A and B that showed significant increase in B factory and inverse significant correlation with air relative humidity. Exposure to high risk of benzene and medium risk of toluene were 10, 14 and 10, 16 percent of workers in factories A and B, respectively. The correlation between occupational exposure and COSHH assessment was significant. occupational exposure to benzene in tire manufacturing personnel was higher than Iranian standard, therefore, improvement of work conditions using appropriate control measures including automated processes and setting of air relative humidity is recommended. COSHH assessment can be used as a simple, cheap and quick method in workers’ health promotion. Control of personnel exposure through ongoing training is imperative.

Backgrounds and Aluminum and in particular its compounds make up a large proportion of the pollutants coming from the aluminum foundry. In several studies conducted on the harmful effects of aluminum in recent decades, it has been identified as a neurotoxic metal. Chronic occupational exposure through inhalation of dust is a common problem in aluminum foundries. Investigation of the exposure of various occupational groups in two aluminum foundry plants to aluminum aerosols. To study the occupational exposure of foundry workers to respirable aerosols of aluminum, personal sampling was conducted from the breathing zone of 63 workers at two foundries in the south of Tehran city following NIOSH method No. 0600. Then samples were treated using NIOSH Institute optimized method No. 7013 and analyzed by high sensitivity graphite atomic absorption.Collected data were analyzed with SPSS V.16 statistical software using an independent sample t-test and ANOVA. The average of aluminum respirable aerosols in A and B factories were 3.21±2.33 and 3.31±2.15 mg/m3 respectively. The one-way ANOVA indicated that occupational exposure among various occupational groups (Similar Exposure Group) in Foundry A had no significant difference (p=0.089), but differences between the exposure of melting and assembling groups in Foundry B are significant (p=0.044). In general, in exposure frequency to aerosols in aluminum foundries, a significant difference was observed between melting and assembling groups (p=0.005) as well among abrading and assembling groups (p=0.02). Most of the exposures to aluminum in foundry workers exceeded the current limits given by ACGIH Institute and the Iranian occupational exposure limits. According to the occupational exposure of both foundries and the absence of differences among variables involved in the exposure, the high exposure of melting and abrading groups compared to assembling groups can be attributed to the nature of these units in the production of aluminum aerosols.

In this project, our aim was to construct a novel expressing vector harboring a new sequence from overlapping region of NS3 gene of HCV from infected Iranian patient.

The partial NS3 (pNS3) gene was amplified by Nested-RT-PCR method using sera of HCV infected patients harboring genotype 1a. After purification and cloning the pNS3 into TA-cloning vector, the best colony was selected based on Blue/White screening and colony-PCR following by confirmation with sequencing and restriction digestion with BglII. The sequenced gene was compared with other reference sequences using alignment softwares. The resultant pNS3 gene subcloned into the expression vector, IRES vector, followed by selection the suitable clones by 2 different colony-PCRs. The gene expression was evaluated using GFP detection, RT-PCR and western blotting techniques after transfection of the IRES-pNS3 vector into the 293 cell line.

After pNS3 sequence amplification by RT-PCR, sequencing results showed high homology among the sequences with other reference sequences. This result also showed that it belonged to genotype 1 of HCV. Colony-PCR showed the insertion of gene into expressing vector with the right orientation. GFP expression, RT-PCR and western blotting confirmed transfection of vector, expression of pNS3 gene and production of its protein in 293 cells respectively.

This novel expressing vector harboring partial region of NS3 gene in compare to full NS3 gene maybe more useful in immune induction by antigen presenting cells due to absence of genes responsible for protease activity of the protein in the setting of HCV vaccine.

Background and ObjectivesHuman immunodeficiency virus (HIV) is the retrovirus that causes the disease known as AIDS. In the current study, we examined the genetic diversity of HIV-1 strains circulating in Iran. Materials and MethodsSequencing and phylogenetic analysis of human immunodeficiency virus type 1 (HIV-1) pol gene is a useful method for subtyping strains of HIV-1. HIV-1 RNA was derived from plasma of 24 infected patients in Imam Khomeini Hospital in Tehran. RT Nested-PCR was developed and the product was extracted from agarose gel. The products were sequenced and phylogenetic analysis by Mega 4 software was constructed. ResultsThe developed PCR method successfully amplified the specific sequence. The sequence and phylogenetic analysis showed subtypes A1 and B in 14 and 10 patients, respectively. ConclusionsThe results indicated intravenous drug users infected with HIV-1 subtype A1, and hemophiliacs infected with HIV-1 subtype B. The results also showed that the subtype A1 is the most prevalent in HIV infected patients. Such information can be useful in intervention strategies and control programs.

Hepatitis G virus (GBV-C) is a single strand RNApositive virus and is a member of flaviviridae family. Infection with the virus is prevalent add has worldwide distribution. Accurate diagnosis of the virus is very important, because of co-infection with other important viruses like HCV, HBV and HIV, it may influence on pathogenesis, disease progression and response to viral therapy.

In the current investigation a sensitive and accurate RT-Nested PCR for isolation and detection of 5'-UTR sequences of Hepatitis G virus (GBV-C) was developed and applied. First genome of the virus was extracted and then, by Reverse Transcription method, cDNA was synthesized. Then, by use of a double stage PCR and two pairs of specific primers, the specified region of the virus genome was amplified and the final product was visualized on agarose gel electrophoresis.

Based on five positive samples, the method was developed and optimized. Then, by use of the developed method, 71 sera samples from chronic HCV infected patients, was checked for Hepatitis G virus (GBV-C). Finally, it was shown that 31(43.6%) patients were positive for Hepatitis G virus.

Based in the results in the current study, it was shown that the developed assay has acceptable performance for diagnosis of Hepatitis G virus infection.Also it was concluded that the infection rate among chronic HCV infected patients is high.