فصلنامه علوم پزشکی دانشگاه آزاد اسلامی
سال بیست و یکم شماره 4 (پیاپی 66، زمستان 1390)

High Resolution Melting (HRM) is a method that analyzes genetic variations such as Single Nucleotide Polymorphisms (SNPs) in PCR amplicons. The STK11 gene encodes a member of the serine/threonine kinase and regulates cell polarity and function as a tumor suppressor gene. The germ-line mutations in this gene are associated with Peutz-Jeghers syndrome. The patients with this syndrome are prone to some types of neoplasm.

In this experimental study, genomic DNA was extracted from the whole blood samples of 56 patients with various gastrointestinal cancers. The nucleotide changes in the entire STK11 gene were analyzed by Real-time PCR and H RM technique.

The nucleotide screening by HRM technique showed two types of SNPs in introns 6 and 7 of STK11 gene in 10 patients. Four patients had C / T substitution [cluster id/dsSNP/rs9282860] with homozygous genotype in intron 6, and six patients showed a C/G substitution [cluster id/dsSNP/rs2075607] with heterozygous genotype in intron 7. The direct sequencing of the fragments confirmed that the results obtained by HRM were 100% reliable.

In this study we found no SNP in exons of STK11 gene. However, two SNPs were found in the introns of this gene. Our results show that the primary screening of the STK11gene by the HRM technique is easily applicable to detect the unknown germ line and somatic mutations in patients with neoplasia at a relatively low cost.

Our previous studies revealed an inhibitory effect of ICD-85; a venom derived peptide has been extracted by researchers of Razi vaccine and serum research institute, on breast cancer cell line (MDA-MB231). The present study was undertaken to evaluate the anti proliferative effect of HL60 cells. The effect of ICD-85 on proliferation of HL-60 cancer cells was determined using the MTT assay. A quantitative cytotoxicity assay based on the release of lactate dehydrogenase (LDH; EC 1.1.1.27) was performed 24 h after exposure to various concentrations of ICD-85. The morphological changes of ICD-85 treated HL-60 cancer cells were observed under electron microscope. ICD-85 induced marked concentration inhibition of cancer cell proliferation with an IC50 value of 0.04µg/ml following 24 h incubation. Electron microscopic findings of ICD-85 treated cells showed nuclear material condensation, endoplasmic reticulum dilation, mitochondria swelling or degradation, increased cytoplasmic vacuoles, reduction or disappearance in cytoplasmic process and decreased nuclear/cytoplasmic ratio. LDH determination of cultured media of HL60 cells exposed to various concentration of ICD-85 revealed no significant changes in LDH levels compared with untreated cells. The results of present study indicated that ICD-85 inhibited the cancer cell proliferation by inducing apoptosis rather than necrosis.

HIV virions with replication capacity are needed for HIV researches, like investigating for new anti HIV agents. Here HIV-1 replication assay was optimized with HIV-1 single cycle replicable (SCR) virions to improve biological safety condition. pSPAX2, pmzNL4-3 and pMD2G plasmids were co-transfected to the HEK293T by using polyfect reagent to produce the SCR HIV-1 virions. Virions were quantified using capture ELISA P24. Different MOI of SCR virions were used for infecting of Target cells (HEK) and the load of the supernatant P24 was monitored days after infection. Single cycle replication assay (SCRA) was developed using kinetic studies data. The P24 load of the infected cells supernatant has linear relation to the beginning infectious MOI. 24 hours post infection with HIV-1 SCR virions the viral particle production was detectable. The highest load of P24 in infected cells supernatant was detected 48 hours after infection. Using this developed method, the 50 and 95 percent inhibitory concentration of (IC95 and IC50) Indinavir and Nevirapine were calculated as 25nM and 50nM. In this study, using SCR HIV-1 virions the SCRA was developed. SCR HIV-1 virions are replicable only for one cycle and this improves the safety of developed assays. The accuracy of assay was examined by quantifying the anti HIV-1 potential of two commercial anti-AIDS drugs and the calculated activity for test agents was equal to previously known amounts.

Cisplatin, as an important anti-cancer drug, is a potent hepatotoxicant. The aim of the present study was to evaluate the protective effect of ethanolic extract of Crocus sativus L. stigma (EECSL.S) against cisplatin-induced hepatotoxicity in the rats. 40 male Wistar rats were randomly allocated into 5 groups (1-Normal control, 2- Toxicant control, 3-Positive control and 4&5- Treatment with extract) of 8 animals each. Cisplatin was injected (0.4 mg/kg) to groups 2-4 daily for 8 weeks, intraperitoneally. Simultaneously, normal saline (10 ml/kg) for groups 1 and 2, silymarin (50 mg/kg) for group 3 and EECSL.S (40 and 80 mg/kg) for groups 4 and 5 were administered through the gavage. At the end of experiment, levels of serum biomarkers of hepatic injury as well as antioxidant activity in liver homogenates of the rats were assessed. Moreover, histopathological observation was assayed at the degree of hepatic injury. In the cisplatin receiving rats, EECSL.S (40 and 80 mg/kg) and silymarin significantly decreased the levels of serum biomarkers of hepatic injury and total bilirubin and significantly elevated the levels of serum albumin and total proteins. In these rats, EECSL.S and silymarin decreased the lipid peroxidation and elevated the levels of antioxidant enzymes in a dose dependent manner. Histopathologically, EECSL.S and silymarin ameliorated cisplatin-induced hepatic injuries and the changes were in agreement with biochemical findings. EECSL.S protects hepatic tissue against cisplatin-induced hepatotoxicity in rats due to its anti-oxidant properties.

Kombucha is used for the treatment of several diseases, including wound healing, in traditional medicine. Due to its effective abilities, such as antibiotic activities, detoxification, and its impact on skin luminosity, we attempted to determine its effect on injured skin in in vivo condition. 30 male NMRI strain were randomly assigned in control (injured skin), sham (exposed with sweet tea) and experimental (received Kombucha) groups. A 3mm diameter wound with complete skin thickness was made on the back of each mouse. Treatment with Kombucha was applied three times a day for 18 days. At the end of study, histological study was carried out on the under-treatment tissues. Significant decrease in wound diameter was observed in experimental group compared to control group (P<0.001). Furthermore, significant increased epidermal thickness and decreased derm and hypoderm thickness were seen in experimental group compared to control group (P<0.05). Hair follicle diameter was significantly increased in experimental group compared to control group, while skin thickness diameter was in coordination with control group. External use of Kombucha on open wound skin with complete skin thickness effectively accelerates wound-healing procedure and also increases hair follicle diameter and length in in vivo condition.

Nicotine is an ingredient of cigarette smoke that has a lot of psychological and physical effects on addicted persons. The aim of this study was to evaluate nicotine effects on functional and histological alteration in thyroid gland that has important role on metabolic activities.

In this experimental study, 40 mature male mice (25-30 gr) were divided to one control group and 3 experimental groups that received nicotine with dosage of 100, 200 and 400 µg/kg via oral gavage once a day for 60 days. Then, T3, T4 and TSH serum levels were measured and histomorphometric evaluation of thyroid gland was done.

Significant decrease in body weight and thyroid follicular epithelial height and significant increase in serum level of T3 were noted.

This study showed that nicotine can cause alteration on metabolic activities of thyroid gland via the increase of T3 serum level and the decrease of follicular epithelial height.

Friedrich ataxia (FA) is an autosomal recessive disorder. Cause of about 2-4% of disease is a GAA triplet repeat expansion in the one allele and carries a point mutation as the other allele. This study was performed to investigate exons of FXN gene to find point mutations for the first time in Iran. In this descriptive study, 50 patients suspected to FA who referred to Special Medical Center were investigated. Genomic DNA was investigated by different PCR methods, including PCR for intron, Long PCR and PCR for exons of FXN gene. Then, products were sequenced and finally sequences were analyzed by related software. C to G nucleotide in intron 2 nt:825954, and T to C in intron 3 nt:832729 of FRDA gene were observed by sequencing method. Nucleotide G insertion was detected in exon 5a nt: 822225. Our study showed that diagnosis of FA is not simple because of clinical overlapping with other ataxia, some mutations in intron maybe affect on the disease which need more examination, and because of consanguinity marriage in Iran, some patients with homozygote mutation may show FA phenotype.

The aim of this study was to evaluate the Enterococcus role in treatment of infectious diseases produced by Staphylococcus aureus by producing of Tyrothricin antibiotic. After the culture of Enterococcus faecalis (PTCC 1394) in the BHI agar, centrifuging and isolation of microbial mass, antimicrobial compound of this strain was purified by dialysis and molecular weight was estimated by SDS-PAGE electrophoresis. Antimicrobial effect and minimum inhibitory concentration of antibiotic was evaluated. Molecular weight of antimicrobial compound was approximately 66 KDa. It was similar to molecular weight of tyrothricin as antimicrobial compound of Enterococcus. After dialysis, activity unit, specific activity, purification factor increased and total activity, amount of total protein decreased. The most antimicrobial effect and the lowest inhibitory concentration produced by tyrothricin were against Staphylococcus aureus. Enterococcus faecalis (PTCC1394) can produce antimicrobial compound with broad specterum, so it can play important role in treatment of angina and intestinal inflammations.

Error in the process of diluting the drugs can lead to administration of inappropriate amount of drugs. Therefore, awareness to the dead space of insulin syringes, which are commonly used for diluting the drugs, is important. In this descriptive study, the dead space volumes of 86 samples from three different syringes made in Iran were measured by using atomic absorption of calcium ions in atomic absorption spectroscopy Varian model Specter AA.200. The dead space of SOHA-syringes® (non removable needle 1 ml insulin syringe) was 5.3±2.2 µl, which was significantly less than its counterparts in V MED-syringe® (removable needle 1 ml insulin syringe with T-shape plunger seal) with dead space of 21.05±9.17 µl and Atlas-syringes® (removable needle 1 ml insulin syringe) with dead space of 54.85±13.15µl. SOHA-syringes with non removable needles are the best for medication dilution in neonatology wards. Among syringes with removable needles, V MED-syringes are preferable to Atlas-syringes because of the smaller dead space.

Mental health status of students has important and efficient role in their social and educational performances. The aim of this study was to evaluate mental health status among the new student in Tehran medical branch, Islamic Azad University. This is a descriptive and causal comparative study in which 465 new students in different medical courses i.e. midwifery, medicine, nursing, laboratory science, environment health and family health were evaluated. The MMPI (Minnesota Multiphasic Personality Inventory) along with demographic were used to collect the data. There was a significant association between depression, hysteria and paranoia with mother and father education. Frequency of depression, hysteria, hypochondriasis, social deviation, mental weakness and schizophrenia were statistically different in various courses. Totally, 19.5 percent of the new student has a form of psychological disorders. Based on mental health status, mental disorders and the relation between certain parameters with mental disorders in new students, It is compulsory such type of research for new entry student of different years to be performed, and counseling center should be active for counseling and educational class to be assigned to the difficulty over come.

Mental health is one of the most important factors that influence aptitude growth of the university students. The purpose of the present study was to assess mental health and some related factors among students of Sabzevar Universities. This cross-sectional study was carried out on 270 students of Universities of Sabzevar in 2010. The subjects were selected using cluster random sampling. The data were collected using GHQ-28. The Cut off point of total score (0-84) was set as 23 and in each item (0-21) was set as 6. In addition, we asked all subjects about social support and parent relationship. The data were analyzed by SPSS version 15.0 software using descriptive and analytical statistics, including t-test, ANOVA and Pearson correlation. 105 (39%) students were suspected to have physical or psychological disorders. Frequency of social dysfunction disorders, depression, physical disorders and anxiety were 57.8%, 35.9%, 32.3% and 23.7%, respectively. Frequency of social dysfunction disorders and depression was more prevalent among males and non-domestic students. Also, mean score of physical disorders and depression was higher in students who their parents were separated or their relationships were with the contention (p<0.05). There was a negative correlation between social support to stress(r=-0.134, p<0.05) and depression(r= -0.189, p<0.01). Considering the high frequency of suspected cases of mental disorders in students, more attention to students’ mental health problems and establishment of counseling centers at Universities are necessary.

Arthritis is a most common manifestation of lyme disease, which is caused by Borrelia burgdorferi. The birds may be carriers of disease-carrying ticks from Asia to Iran. The aim of this study was to evaluate frequency of lyme arthritis in patients with unknown subacute arthritis. This cross-sectional study was evaluated 180 patients with unknown subacute arthritis. Patients were asked about age, sex, foreign travel history, jungle travel history, animal contact, tick bites. Then, IgG, IgM Ab ELISA test were done. In patient with positive IgG or IgM ELISA test, Western Blot test was done. SPSS was used to analyze data. 20 (11.9%) patients had positive ELISA serologic test, in which 12(60%) had IgM Ab, 5 (25%) IgG Ab and 3 (15%) both IgM Ab and IgG Ab. Western Blot was positive in 2(1.1%) and equivocal in 5(2.8%) patients. The results of this study showed that lyme disease should be considered in each patient with unknown subacute arthritis in Iran.