Journal of

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Introduction: DNA extraction methods are very important for genetics research and diagnostic tests. In These methods are also important for detecting genetic diseases or in cancers for investigating genetic changes during cancer progression or treatment. Therefore, selection of the best method for DNA extraction from different samples such as bone marrow (BM) or peripheral blood (PB) and their slides is very important. Methods: In the present research, DNA was extracted from 5 different samples including 1-PB, 2-BM slides stained by Gimsa method, 3-Gimsa stained PB slides from archives, 4-new Gimsa stained PB slides and 5-non stained new PB slides by 3 different methods salting-out, boiling and phonal chloroform method. In all of the groups, three DNA parameters were investigated 1-OD (Optimal Density), 2-DNA concentration and 3-Outcome (PCR results). Results: The best DNA quality was achieved by salting-out method (OD=1.74), while the worst quality was by the boiling method (OD<1.0). The DNA quality of all the samples was similar in the salting-out and phenol chloroform methods. Regarding DNA quantity, the best result was from boiling method (6.7µg / ml). The least amount of DNA was obtained by the phenol chloroform method and salting-out method also resulted in the least quantity of extracted DNA. Regarding the outcome of DNA extraction or the PCR results, all the three methods showed 100% positive results for peripheral blood samples, while boiling method had the best outcome for BM slides, archive stained PB slides, new stained PB slides and non-stained PB slides (100%, 88%, 84% and 72% respectively). Discussion: The present research indicated that except non stained PB slides, the DNA extraction from all other samples showed very good results. In addition, the research showed that there is no difference in DNA extraction of new or archive slides.

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Introduction: Medicinal plants are valuable natural resources that are nowadays known as safe pre-materials for drug industries in the advanced countries. Black caraway is one of the most important and economical medicinal plants that can be grown naturally in some regions of Yazd province. The essence of the plant are used in treatment of diarrhea, dyspepsia, fever, flatulence, stomachache, hemorrhoids, hiccoughs, as an antihistaminic and more numerous uses in medicine industries. This plant shows several therapeutic effects on digestive and urinary tract disorders and is well known as an anti-convulsant, anthelmintic, anti-asthma and antidyspnic drug in Iranian traditional medicine. Bunium Persicum oil is capable of suppressing the initial stage of an inflammatory process. Also, this plant is used for culinary purposes and for flavoring foods and beverages. Methods: In this research, the fractionization of seed essence was done by GC method and the compounds were recognized. The DPPH test was used for estimating anti oxidant effects and the Follin-Ciocalteu method was used for estimating quantity of phenol compounds. Results: The results revealed that- γTerpinene had the highest percentage in the essence of the native black caraway of Yazd province. Also, the antioxidant test showed the high antioxidant effect of the native black caraway of Yazd province with an IC50 of 2.85 µg.mg-1 and the phenolic component percentage of 117.09mg.g1. Conclusion: This result was much higher than previous similar studies about black caraway.

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Introduction: The importance of scientific research in prevention of drug addiction is obvious. The main site of drug action is the brain. Mu receptor is the first place of drug action. In this study, the rate of mu receptor gene single nucleotide polymorphism (SNP) was compared in addicts and non-addicts. Methods: In this study AA, AG and AA polymorphism of mu receptor gene was investigated in 47 addicts and 30 non-addicts (controls). DNA was extracted by salting out method from peripheral blood and the PCR-Arms was performed. Results: In control group, 66.7% had AA polymorphism while 68.1% of addict group showed this polymorphism. The AG polymorphism was 30% and 27.7% in control and addict groups, respectively. The GG polymorphism was 3.3% in control and 4.3% in addict group. There were no significant differences between the two groups. Discussion: Our results indicated that there is no relationship between the mu receptor gene SNP and addiction. The other SNPs in this gene and other genes must be investigated. In addition, it seems that other factors such as environmental factors could play a role in addiction.

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Introdution: Presence of mentally retarded children as a source of pressure can jeopardize the general health of parents, especially mothers. The range of effect depends on the recognitive evaluation and the individual. The aim of this study was to investigate the effect of coping-therapy on mental health of mothers with genetically and non genetically mentally retarded children referring to Yazd clinical center. Methods: This study was semi experimental and included 40 mothers with mentally retarded children studying in schools supported by the welfare organization of Yazd in 2009- 2010 and were selected by available sampling method. They were divided to two groups case and control. Before any therapy, all of the mothers answered a general health questionnaire(GHQ28). In the next step, coping-therapy was performed on the case group. In the end, all of the mothers answered the same questionnaire(GHQ28) and data were analyzed by covariance method and t test. Results: The research indicated that coping-therapy has a positive effect on the mental health of mothers with genetically mentally retarded children. This effect is similar on mothers of children with non genetically mental retarded children. Coping-therapy decreases the somatic signs of depression in mothers and improves their sleeping and social efficacy. There was no association of age and educational level of mothers with coping-therapy. Conclusion: Coping-therapy can improve the mental health of mothers of both genetically and non genetically mentally retarded children

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Introduction: Undoubtedly, reproduction is a necessity for survival and successful pregnancy is an immunological paradox. In the present study, we investigated the proportional changes of CD4+CD25bright T cells, CD4+CD25dim T cells in peripheral blood in unexplained recurrent spontaneous abortions (URSA) and compared it with normal pregnant women by antibody monoclonal method. Methods: The study group comprised of women with miscarriages of unexplained etiology who had normal karyotypes, anticardiolipin antibodies, prolactin levels and normal spousal spermograms. They did not have polycystic ovaries and also did not receive any drugs at the time of the study. PBLs lymphocytes were isolated, then FITC-conjugated and anti-CD4 and PE-conjugated anti-CD25 antibody levels were measured. Then results of the study and control group were analyzed and compared. Results: The absolute number of CD25 bright cells in the CD4‏+T cells in peripheral blood was statistically significantly lower in the study group as compared to the control group(P=0.000). The absolute number of CD4+CD25dimT cells in peripheral blood was statistically significantly higher in the study group as compared to the control group (P=0.000). Conclusion: As decrease in the number of CD4+CD25+Tcells or their functional deficiency may be linked with miscarriage, CD4+CD25+‏ Tells could serve as a novel biomarker for monitoring in URSA patients, but more studies are needed in this field.

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Introduction: Immunological processes are most important in recurrent spontaneous abortion(RSA). According to studies T lymphocytes and natural killer cells(NK cells) are two effective cell groups in RSA. The aim of this study was to investigate the percentage and absolute number of natural killer(NK) cells in women with RSA with unknown etiology. Methods: 24 Women with a history of recurrent pregnancy losses with unknown etiology. We compared the percentage of peripheral blood NK cells with a group of fertile patients. Lymphocytes from peripheral blood were isolated by ficoll paque density centrifugation. Lymphocytes were stained using anti CD56 and(FITC)-anti CD16 and CYQ-CD3 monoclonal antibodies for identification of NK cells and was used anti CD56 and(FITC)-anti CD69 for detection of activated NK cells. We used BD FACS calibure flow cytometry for data analysis. Results: On the basis of the obtained results, absolute number of CD16+56+ cells showed significant increases in Recurrent spontaneous abortion(RSA) in comparison with control group(P= 0.43). Also absolute number of CD16+56bright cells had significant increase in RSA(P=0.00). There was no significant difference(P= 0.08) of CD16+56dim cells between RSA and control group. In RSA, the absolute number of CD69+cells significantly increased(P=0.02). Also, results showed significant increase in the absolute number of CD56+/CD69+ cells in RSA(P=0.04). Conclusion: The results suggested that the higher percentage of NK cells in peripheral blood of RSA patients compared to control group may indicate the same increase in number and cytotoxicity of uterin NK cells.

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Introduction: GSTM and GSTT are subclasses of glutathione s-transferase that is present on human sperm surface and plays an important role against oxidative stress. This study aimed to investigate the polymorphisms of GSTT1 and GSTM1 in regard to sperm parameters. Methods: This case-control study involved 46 men with varicocele and 48 men without varicocele. Semen analyses were carried out according to WHO guidelines. Blood DNA was extracted using salting out procedures. Polymorphism of GSTT1 and GSTM1 genes was determined trough multiplex-PCR respectively. Results: Frequencies of GSTM1 null genotype in men with varicocelc and men without varicocele groups were 60.9 and 41.7 respectively. There were no statistically significant differences between Gstm1 null and positive genotype in two groups (p>0.05). Frequencies of gstt1 null genotype in case and control groups were 47.8 and 50 respectively. There were no statistically significant relationship between gstt1 null and positive genotype in two groups (p>0.05). Conclusions: Deficiency of enzyme activity in gstm1 null genotype did not affect morphology as well as slow and quick progressive of sperm but caused the significant decrease in count of sperm between gstm1 null and positive genotype. In the case of gstt1, gstt1 null genotype did not affect sperm parameters that may be related to compensate activity of other genes in this super family.

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Introduction: Previous studies have shown altered levels of n-3LCPUFA in the pathophysiologcal conditions such as preeclampsia .Also elevated expression of sFlt-1 in preeclampsia plays a major role in the pathogenesis of this serious disorder especially in reduced placental oxygenation. The present study examines the hypothesis that Eicosapentaenoic acid (EPA 20:5), an omega-3 long-chain polyunsaturated fatty acids (n-3LCPUFAs), may attenuate sFlt-1 gene and protein expression in JEG-3 cells treated with induced hypoxia-like conditions by (DMOG) -induced hypoxia-like conditions. Methods: JEG-3 cells were pretreated with DMOG incubated with EPA. Protein expression of sFlt-1 was measured by enzyme-linked immunosorbent assay (ELISA). Messenger RNA expressions of sFlt-1 was determined by and RT Real Time-PCR. Results: Our results showed that incubation of JEG-3 cells with DMOG cause a significant elevation in mRNA levels and protein secretion of sFlt-1(P < 0.05). In contrast, EPA decreased the mRNA expression and protein secretion of sFlt-1(P < 0.05). Also mRNA expression and protein secretion of sFlt-1 inhibited cells treated by both EPA and DMOG (P=0.261, P =0.077 respectively). Conclusion: These findings confirm previous studies that hypoxia caused elevation in sFlt-1 gene expression and protein secretion. Also our studies reveal that effects of n-3 fatty acids in restraining preeclampsia complications may be mediated by suppressing the gene expression and protein secretion of sFlt-1 under hypoxia conditions. This data provide evidence that n-3 LCPUFA can exert its effects through inhibition of the HIF pathway

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Introduction: Infertility is described as the inability to get pregnant after one year of unprotected intercourse. About half of infertility cases are because of male factors. Idiopathic azoospermia or severe oligozoospermia caused by genetic alterations is a significant part of male infertility. A key step of spermatogenesis is crossover events during meiotic reciprocal recombination. MLH3 protein has a crucial role in meiotic recombination and in spermatogenesis. We evaluated this function of MLH3 protein by examining the contribution of functional polymorphism in MLH3 (C2531T) to the risk of male infertility. Methods: We studied this polymorphism in 110 infertile male with idiopathic azoospermia or severe oligozoospermia, and 110 fertile men with normozoospermia as a control group. MLH3 C2531T polymorphism was analyzed using the tetra-amplification refractory mutation system-PCR (4P-ARMS-PCR) method. Results: Genotypes CC, CT and TT of the MLH3 gene presented frequencies of 13.6%,59.1% and 27.3%, respectively, in the men with idiopathic azoospermia or severe oligozoospermia and 37.3%,53.6% and 9.1% in the control group (p<0.001). Conclusion: The data suggest that the MLH3 C2531T polymorphism can be associated with risk of male infertility. The research data showed that presence of the polymorphic allele T leads to an increased risk of 2.35 times (OR =2.35, 95% CI =1.57-3.51 p<0.001) to develop infertility in relation to the normal control group. Therefore, the MLH3 gene polymorphism may be genetic determinant for defective spermatogenesis in the humans.

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Introduction: Age estimation plays a critical role in dentistry especial in orthodontics, pediatric Dentistry and other majors such as archaeology, etc. The aim of this study was to evaluate the accuracy of dental age estimation in 5-16 years-old children and adolescents in Gilan using Demirjian method. Methods: This retrospective cross-sectional study was carried out on 314 panoramic radiographs of 212 girls and 102 boys, referring to Gilan dental clinics. Simple sampling method was used. Based on Demirjian method two dental radiologists calculated the dental maturity score (DMS) by assessing the seven left mandibular teeth and estimated the dental age for each case. The chronologic age of each patient was obtained from demographic data. Finally, the correlation between estimated dental age and chronologic age was determined by Pearson’s correlation coefficient and paired t-test using SPSS statistical software (α = 0.05). Results: Mean dental age in all the subjects was estimated 0.02 year less than the chronologic age. In addition, there was no significant difference between estimated and chronological ages (p value = 0.71). Pearson’s correlation coefficient between estimated and chronological ages was 0.73, indicating statistical significance (pvalue < 0.001). Conclusion: Based on the results of the present study, Demirjian method is sufficiently accurate to estimate dental age of children in Gilan and can be used acceptably.

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Introduction: The physiological changes in pregnancy lead to the psychological pressures. Therefore, there is a need for applying cognitive behavioral and emotional confronting to encounter these problems. The present research investigated the effect of coping therapy on general health of pregnant women with high risk of genetic abnormalities in their fetus.

Methods: The present study was a semi experimental research. Pre and post tests were used to investigate coping therapy between 30 pregnant women who were referred to Khatomolanbia Genetic Clinic, Yazd, Iran. All the women had pregnancy screening test with high risk of genetics abnormalities in their fetus. They were divided randomly into two groups of case and controls. The test of GHQ was performed in both groups, then the case groups went under 8 sessions of teaching coping therapy each lasting 120 min. After finishing the sessions, post test was performed and analyzing the data using descriptive statistical index and covariance analysis test.

Results: Teaching coping therapy to case group caused improvement in their GHQ mark, and this change was significantly different from the change in the GHQ mark of control group. In addition, there was a significant decrease in anxiety, depression and physical signs and an improvement of social function in case group compare to the control group.

Discussion: Teaching coping therapy can improve the general health of pregnant women with high risk of genetic abnormalities in their fetus. Therefore, presenting educational courses to the women can improve their general health indices in addition to preventing the probable effects of stress on fetus.

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Introduction: Cancer is one of the most harmful disease throughout the world. Doxorubicin is an anti-cancer agent, used in the treatment of various types of the cancer such as bone cancer. There are several adverse effects related to clinical usage of Doxorubicin for long time. The present study aimed to investigate the reducing side effects and enhancing the therapeutic effect by liposomal carrier.

Methods: Liposomes containing DPPG and cholesterol with the molar ratio of 70:30 with the Doxorubicin were synthesized by pH- gradient method. The average diameter of nanoparticles and surface charge was determined by Zeta-Sizer instrument. The amount of drug loaded and drug-released was determined using dialysis. The surface morphology and internal lamella was evaluated by TEM and SEM.

Results: The average size of liposomal Doxorubicin obtained using Zeta-Sizer was 126 nm. The encapsulation efficacy of liposomal Doxorubicin was 89%. The total amount of drug release during 48 hours in acidic medium studied by dialysis technique was 46%.

Conclusion: In this study, investigation of loading Doxorubicin into nano-liposome with the slow- released kinetic was carried out to improve the solubility and bioavailability of Doxorubicin in order to delivery to osteosarcoma cell line.

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Introduction: Since the disability may be associated with functional impairments and the fact that exercise can reduce disability and maintain the function of individuals, so, we decided to conduct a study on the effects of resistance training on shoulder strength, pain and function in wheelchair users with impingement syndrome of shoulder.

Methods: In this randomised clinical trial study, In this randomized controlled intervention study, 20 wheelchair users with spinal cord injury suffered from shoulder pain (the mean age (39/3±6/4)) were chosen and divided randomly into two groups: the control (age:37/5±5/81, height:82/25±4/20, weight:77/25±12/98) and experimental groups (age:42/1±8/11, height:84/95±4/7, weight:74/23±11/39). Before performing the training programs, the pretests of the level of muscle strength, pain and function were measured for both the control and experimental groups. The experimental group performed selected exercises for 8 weeks, 3 sessions a week, each session lasted 60 minutes. After performing the training course, post-tests were administered in similar circumstances in both groups.  Data were analyzed by using covariance and dependent t-test.

Results: The results showed that the experimental group had a significantly improvement in the strength of external shoulder rotator muscles (from 7/66±1/38 to 10/71±1/73), pain (from 52/48±7/96 to 30/31±5/24) and the shoulder function (from 55/49±6/66 to 72/88±6/52) (P≤0/05).

Conclusions: This study showed that strength training has improved the strength, pain and shoulder function in wheelchair users, so participation in exercise training programs can be suggested for these persons.

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Introdution: Endometriosis is one of the most common gynecologic disorders and shows significantly elevated prevalence in industrial regions. Additionally, a possible genetic predisposition is assumed for the disease. Glutathione S-transferases (GSTs) are enzymes participated in the metabolism of many human disease-causing mutagens, carcinogens and environmental pollutants. A functionally significant A to G transition in GSTP1 gene causes substitution of in the codon 105 can influence the enzyme activity. The aim of the present study was to investigate association of GSTP1 polymorphism and endometriosis in women from central and southern Iran.
Methods: In this case-control study, after obtaining informed consent, samples were obtained from 101 endometriosis patients and 126 healthy controls. Genomic DNA was isolated from peripheral blood cells and genotyping was performed using polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) analysis. Data were compared in both groups by using Pearson chi-square and Hardy-Weinberg equilibrium tests.
Results: Results showed no significant association between GSTP1 Ile105Val Polymorphism and endometriosis susceptibility (P = 0.370). Frequencies of the AA, AG and GG genotype of GSTP1 gene polymorphism in the patients were 42.6%, 47.5% and 9.9%, while the frequencies in the controls were 49.2%, 45.2% and 5.6%, respectively.
Conclusion: According to our study, GSTP1 Ile105Val polymorphism appears to be not associated with the risk of endometriosis in the studied population. However, additional studies, especially with larger sample size are needed to validate these findings.
 

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Introdution: In the present study, Lipofactamine 2000 was used as a cationic liposome for miR-101 transfection in order to investigate its cytotoxicity and its effect on the expression of ubiquitin ligase HECTH9 in acute myeloid leukemia cells (AML).
Methods: MiR- 101 was transferred to KG-1 cells (myeloid cells) and HBMF-SPH (healthy bone marrow cells) using lipofectamine 2000 as a nano carrier. Then, using the MTT test, the 48-hour cell toxicity in both cell lines was evaluated. The effect of this miRNA on the expression of HECTH9 gene (ubiquitin Ligaase E3) was evaluated using qRT-PCR technique.
Results: The findings of this study showed that Lipofactamine alone was not toxic to any of the cell lines, but lipofectamine-containing miR-101 (Lipo / miR-101) in KG-1 cells produced the highest toxicity compared to other treatments. The results of qRT-PCR test showed that Lipo / miR-101 treatment in KG-1 cells caused the highest expression in HECTH9 gene at the mRNA level.
 
Conclusion: Lipofactamine, as a cationic liposome, can effectively transfect miR-101 into the cells and can cause miR-101 to specifically display its antitumor effect by increasing the expression of HECTH9 and regulating pathways of mitochondrial apoptotic pathway. Therefore, miR-101 can be used as a potent tumor suppressor and an effective therapeutic agent for gene therapy in the patients with AML.

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Introdution: The aim of this study was to investigate the physico-chemical properties, surface charge and cytotoxicity of nanocomposite formulations for the design of lipid nano-systems with maximum loading for nucleic acids in bone marrow cancer.
Methods For this study, six types of liposomes with different compositions were synthesized using DPPC, cholesterol, DSPE-mPEG (2000) and DOTAP. Then, nanosystems were evaluated for particle size, surface charge, cytotoxicity and loading rate of miRNA in two MG-63 and SaOs2 cell lines, which are human cell bone marrow cell lines.
Results: All nano-systems were monodisperse. Among the systems, the F6 formula has the lowest toxicity and the highest load of miRNA-143, due to the presence of a suitable amount of DOTAP and PEG phospholipid in the liposome structure.
Conclusion: Liposomal formulation with a suitable percentage of DOTAP cationic phospholipid can be used as a successful carrier for the transport of miRNA in the gene therapy process for the treatment of various cancers, especially metastatic types.

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Introduction: In the recent decades, the prevalence of Klebsiella pneumoniae beta-lactamase producing strains has a significant role in nosocomial infections. The aim of this study was to determine the prevalence of different types of CTX-M enzymes among clinical isolates of K. pneumoniae.
Methods: In this descriptive cross-sectional study, during six months, K. pneumoniae strains were isolated from laboratory ward of Babol educational hospitals and referred to microbiology laboratory of Babol University of Sciences for final confirmation. Thereafter, antibiogram pattern analysis and the frequency of CTX-M1 and CTX-M15 genes were evaluated. Finally, the results were analyzed by SPSS version 24 software.
Results: During six months of this study in 2017, 65 K. pneumoniae and also other bacterial strains such as Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii were collected from laboratory ward of Shahid Beheshti and Ayatollah Rouhani Hospitals. By disc diffusion method, the highest resistance belonged to Amoxicillin (91%), Cefoxitin (45%), Piperacillin (45%) and the lowest belonged to Amikacin (24%) and Imipenem (0%). On the other hand, the percentage of CTX-M1 and CTX-M15 genes were also found to be 62% and 69%, respectively.
Conclusion: Regarding to the high prevalence of strains containing broad-spectrum beta-lactamase enzymes and due to the antibiotic resistance among K. pneumoniae strains isolated from clinical specimens and also by considering this issue that the current study was the first research for evaluating the frequency of these genes in Babol educational hospitals therefore, it is necessary to adopt appropriate drug regimens to reduce antibiotic resistances

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Introdution: The present study was conducted to evaluate the effect of miR-143 on the expression of Bcl2 gene at the level of mRNA, cell cytotoxicity in the SaOs-2 cell line (Osteosarcoma) and Hum 63 (Human primary bone cells).
Methods: In this experimental study, miR-143 was introduced into the SaOs-2  and Hum 63 cell by the lipofectam 2000 system. Then, the effects of miR-143 on cytotoxicity (72 hours) and expression of Bcl2 gene were measured by the MTT tests and Real Time Pcr protocols, respectively.
Results: The results showed that the lipofectamine 2000 system had no toxicity in both Hum63 and SaOs2 cell lines by itself. SaOs2 cancer cells were more susceptible to miR-143 and Lipo / miR-143 compared to Hum63 cells. The highest reduction in SaOs2 cells was observed after the treatment with Lipo / miR-143 (p <0.05). Furthermore, according to the results, transfection of miR-143 by lipofectamine 2000 into the cell, compared to cell-treated alone by miR-143, significantly reduced Bcl2 expression (p <0.05).
Conclusion: The lipoprotein 2000 system can be effective in absorbing miR-143 into the cells. By targeting various cellular pathways, including the expression level of the Bcl2 gene, miR-143 can be effective in the treatment of bone cancer (Osteosarcoma).

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Introdution: Nanotechnology introduced new methods to chemotherapy drugs delivery into cancer cells to reduce the side effect of drugs it increases the quality of cancer treatment and reduces the side effects of chemotherapy. . In this study, different lipid formulations of nucliposomes containing curcumin was prepared by thin-layer method and evaluated for chemical-physical evaluation on MCF-7 cell line.
Methods: This study was a descriptive-analytic. Various formulations of liposomes prepared from Span 60, SPC and cholesterol. Curcumin was loaded in the liposomes. Final liposome formulation selected and PEGylated. The Optimal formulation characterized by DLS, FT-IRT, Fe-SEM and SEM. Statistical analysis was performed with Prism6 software.
Results: The average size of liposomes, encapsulation efficacy of liposomal loaded curcumin and Zeta potential was about 63.9 nm, -41mV and %95.58, respectively. Liposomes exhibited the slowest release rate in normal and turmeric cell culture condition. To compare the function of free curcumin in similar concentrations, liposomal curcumin formulation was %63 toxic to MCF7 Cell line.
Conclusion: The present study shows that an optimally optimized nanoliposome with curcumin having appropriate physicochemical properties, improving drug toxicity in cancerous cell lines and increasing drug stability can be a good vector for drug delivery to breast cancer cells.
Conclusion: The present study shows that an optimally optimized nanoliposome with curcumin having appropriate physicochemical properties, improving drug toxicity in cancerous cell lines and increasing drug stability can be a good vector for drug delivery to breast cancer cells.