mohammad reza sarookhani

In this study, using evaluation of the expression of two specific proteins of endoplasmic reticulum (ER) stress, GRP78 and CHOP in the striatum and substantia nigra (SN) in the 6-hydroxydopamine (6-OHDA) animal model of Parkinson’s disease (PD), the role of this stress was evaluated in the progress of Parkinson's disease (PD). 

6-OHDA was injected into medial forebrain bundle. Behavioral tests were carried out in the second, forth, sixth and eighth weeks after the toxin. In the eighth week, the brain of some rats was perfused and immunohistochemical (IHC) assessments were performed to evaluate the survival of dopaminergic neurons in SN and also the expression of GRP78 and CHOP in striatum. The brain of other rats was freshly removed and the expression of GRP78 and CHOP in SN was evaluated using western blotting.

The severity of behavioral symptoms increased progressively in 6-OHDA- treated rats and reached to maximum in the eighth week. IHC assessments revealed that more than 80% of dopaminergic neurons in SN were lost in these rats. These assessments also showed that only 5% of the cells in striatum of control rats expressed GRP78 and CHOP. On the other hand, about 42% of these cells in 6-OHDA- treated rats expressed these proteins. Furthermore, expression of GRP78 and CHOP in SN of 6-OHDA- treated rats increased 400% as compared to control rats.

ER stress involves in progress of 6-OHDA-induced parkinsonism in rat indicating this stress may have a role in progress of PD in human beings.

Multidrug resistance (MDR) is a major obstacle in the successful chemotherapy of ovarian cancer. Inhibition of P-glycoprotein (P-gp), a member of ATP-binding cassette (ABC) transporters, is a well-known strategy to overcome MDR in cancer. The aim of this study was to investigate the efficiency and ability of CRISPR/Cas9 genome editing technology to knockdown ABCB1 gene expression in adriamycin resistant (A2780/ADR) ovarian cancer cell line and evaluate the sensitivity changes to doxorubicin.
Three single-guide RNAs (sgRNAs) targeting the fourth and fifth exons of human ABCB1 gene were designed in this study. Expression level of ABCB1 was detected using quantitative real time PCR (qRT-PCR) after co-transfection of all three sgRNAs into A2780/ADR cell line and subsequent antibiotic selection. Drug sensitivity to doxorubicin was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
The results showed that CRISPR/Cas9 system could significantly reduce the expression of P-gp. The dramatic decline in ABCB1 gene expression was associated with increased sensitivity of cells transfected with sgRNAs to doxorubicin.
Based on the results of this study, it is concluded that the CRISPR-based systems, used in the present study, effectively down-regulated the target gene and acted as an ideal and cost-effective tool for gene editing of A2780/ADR cell line resulting in restoration of nonmalignant phenotype.

We aimed to determine the relationship between serum glutathione peroxidase and febrile seizure.
In this case-control study, 43 children with simple febrile seizure (case group) were compared with 43 febrile children without seizure (control group) in terms of serum glutathione peroxidase level, measured by ELISA method. This study was conducted in Qazvin Children Hospital, Qazvin University of Medical Sciences in Qazvin, Iran in 2012-2013. The results were analyzed and compared in two groups.
From 43 children 24 (53%) were male and 19 (47%) were female in children with simple febrile seizure, and 26 (60%) were male and 17 (40%) were female in febrile children without seizure (control group) (P=0.827). Serum glutathione peroxidase level was 166 U/ml (SD=107) in the case group and 141 U/ml (SD=90.5) in the control group of no significant difference.
There was no significant relationship between serum glutathione peroxidase and simple febrile seizure. Thus, it seems that glutathione peroxidase, an essential component of antioxidant system, does not play any role in the pathogenesis of simple febrile seizure.

Micro-RNAs (miRNAs) are a class of posttranscriptional regulators that play crucial roles in various biological processes. Emerging evidence suggests a direct link between miRNAs and development of several diseases including type 2 diabetes (T2D). In this study, we aimed to investigate the effect of predicted miRNA and target genes on insulin resistance.
This experimental study was conducted on the C2C12 cell line. Using bioinformatics tools miRNA-135 and two respective target genes-insulin receptor (Insr) and vesicle associated membrane protein 2 (Vamp2)- were selected as potential factors involved in insulin resistance process. Levels of glucose uptake miRNA expression and respective gene targets were determined after cell transfaction by miR-135.
It was determined that Insr gene expression was significantly down-regulated in miR-135 transfected C2C12 cell line (P≤0.05). Interestingly; these transfected cells have shown a significant difference in glucose uptake incomparision the positive control cells, while it was similar to the insulin resistant cell line (P≤0.05). In contrast, no significant alteration of Vamp2 gene expression was observed.
Our data indicated no change on the Vamp2 expression level after miRNA transfection, while expression level of Insr was reduced and miR-135 expression was contrarily increased leading to poor stimulation of glucose uptake through insulin, and development of insulin resistance phenotype in C2C12 cell line.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that play pivotal roles in many biological processes such as regulating skeletal muscle development where alterations in miRNA expression are reported during myogenesis. In this study, we aimed to investigate the impact of predicted miRNAs and their target genes on the myoblast to myocyte differentiation process. This experimental study was conducted on the C2C12 cell line. Using a bioinformatics approach, miR-214 and miR-135 were selected according to their targets as potential factors in myoblast to myocyte differentiation induced by 3% horse serum. Immunocytochemistry (ICC) was undertaken to confirm the differentiation process and quantitative real-time polymerase chain reaction (PCR) to determine the expression level of miRNAs and their targets. During myoblast to myocyte differentiation, miR-214 was significantly downregulated while miRNA-135, Irs2, Akt2 and Insr were overexpressed during the process. miR-214 and miR-135 are potential regulators of myogenesis and are involved in skeletal muscle development through regulating the IRS/PI3K pathway.

Early diagnosis and treatment of leukemia patients remains a fundamental aim in clinical oncology, especially in developing country. Present study highlights the basic requirements of these patients in Iran. Better understanding of these issues may lead to improve the healthcare standards toward leukemia diagnosis and treatment. This descriptive study included 101 specialists in hematology-oncology and pathology serving in oncology centers. The participants were then asked to fill out a standard questionnaire on the issues around diagnosis and treatment of blood malignancies. According to specialists, unfair distribution of facilities across the country, delayed diagnosis of disease, absence of psychological support for patients, and insufficient financial support were the main reasons of inappropriate diagnosis and treatment in leukemia patients. Our results show that making an amendment to health policies by preparing well-equipped medical centers in all provinces, improving the morale of patients through consultation during the process of treatment, and above all, subsiding leukemia patients'' financial problems will promote the health standard regarding the leukemia diagnosis and treatment in Iran.

Vulvovaginal candidiasis (VVC) is a common problem in women. The objectives of this study were to compare two identification methods، i. e. new PCR analysis (with DNA extracted from sapmles) and conventional culture technique in the detection of Candida species in vulvo-vaginal samples. In an experimental-analytical study، 150 sexually active women participated in this study Vaginal discharge was sampled using two sterile Dacron swabs، of which one used for Direct DNA extraction as well as PCR and the other for culture and phenotypic evaluations. Phenotypic evaluations was performed by germ tube and chlamidospore formation and specially culture in chrome agar medium to detect color of the colonies. PCR was performed by DNA extracted from samples as templates and finally size of Candida specific amplicons were determined. From 150 samples، 87 were positive in culture and 128 were positive in new PCR technique. In culture method، from total 87 Candida species، 73 C. albicans، 12 C. glabrata and 2 C. tropicalis were isolated whereas in new PCR technique، from total 128 candida species، 108 C. albicans، 18 C. glabrata and 2 C. tropicalis were identified. Concordance of the two methods were calculated as 68%.

Early and accurate diagnosis of bacterial meningitis is of critical concern. Optimum and rapid laboratory facilities are not routinely available for detecting the etiologic agents of meningitis. The objective of this study was to compare polymerase chain reaction (PCR) assay with culture for detection of bacteria in central nervous system (CNS) samples from patients suspected to have meningitis. One-hundred CSF samples were obtained and divided into two parts. One part of samples was used for standard bacterial culture and gram staining. The remaining was used for DNA extraction. PCR assay was performed with universal primers for 16S rDNA gene of bacteria. Performance characteristics of the test were determined. The PCR method was able to detect bacteria in all 36 culture-positive and in 38 of 64 culture-negative cases showing sensitivity and specificity of 100% and 40.6% respectively. Positive predictive value was 48.6% and negative predictive value 100%, however, Kappa coefficient showed the correlation of the 2 methods to be at 0.33. There are advantages and disadvantages in performance characteristics of the conventional CSF culture and universal CSF 16S rDNA PCR. Therefore, it is recommended to use both methods in clinical practice, particularly in suspicious contaminated samples, with presumable presence of fastidious or slow growing bacteria because of antibiotic consumption.

Secondary hyperparathyroidism (SHPT) is one of the serious complications in patients with chronic kidney disease (CKD). This study aimed to identify biochemical alterations of renal bone disease in hemodialysis patients of Qazvin province. In a descriptive study, fasting blood samples of arterio-venul shunt, before starting hemodialysis, were taken from all CKD patients and Ca++, P-- and ALP were measured by colorimetric methods and PTH by IRMA method. Descriptive statistics was used to present data. In 4% of cases there were no abnormalities of mentioned parameters but in 96% of patients one or more parameters were abnormal. The most prevalent abnormality was related to P-- (increased) and the least one to ALP (increased). 51% of patients had raised PTH level (hyperparathyroidism) and higher abnormalities of other biochemical parameters. No differences were seen in the mean of age, duration and number of hemodialysis and also sex ratio of hyperparathyroid patients and all studied patients. The Biochemical and hormonal results revealed a predominance of mild to moderate secondary hyperparathyroidism and renal bone disease in CKD patients, so there is a need to control the disease with specific treatments.