International Journal of Medical Laboratory

89-299

Authors: Zeinab Mahdian1 Ph.D., Fereshteh Amiri2 M.Sc., Zohreh Mohammadi3 M.Sc., Parviz Farzadinia4 Ph.D., Fahimeh Safizadeh5 M.Sc, Zahra Zare6 Ph.D., Rahimeh Rahimi7 M.Sc., Zahra Dehghani8 M.D., P.hD., Fariba Mohammadi Tahroodi9* M.Sc.

Background and Aims: Burn injury remain as a major medical problem throughout the world. This injury is accompanied with inflammatory and wound healing responses. Since royal jelly (RJ) has anti-inflammatory and wound healing activity therefore, the aim of this study was to evaluate the repairing effects of RJ on skin burn- damage. Materials and Methods: In an experimental study, 40 male Wistar rats (8 weeks old) were engaged. The animals were divided into five equal groups. Group 1 was considered healthy control. Group 2 (positive control) was treated topically with Silver Sulfadiazine Cream, group 3 received Eucerin as negative control, and group 4, 5 treated with RJ (10 and 30%). Sampling was performed after observing the second-degree burns on the first, seventh and fourteenth days. Then after 28 days, rats were sacrificed and their skin tissues were used for morphological and morphometric assessments. Results: The results of this study showed that the amount and arrangement of collagen type 1 protein was higher in the RJ treatment groups versus control group. Reconstruction and thickening of the epithelium in RJ treated groups confirmed therapeutic effects of RJ. In addition, RJ increased angiogenesis compared to the control group. The wound's surface area was reduced in the RJ treatment groups compared to the control group. In addition, fibroblast cell proliferation was increased in the groups receiving RJ versus control. Conclusions: It could be concluded that, RJ induce wound healing effects and might be considered as potential treatment option to improve the burn wound healing.

251-257

Authors: Tahmine Zafari1 M.D., Narges Ajilian1 M.D., Atena Mansouri2,3 Ph.D., Ariane Sadr-Nabavi4,5,6 Ph.D., Seyed-Alireza Esmaeili7,8 Ph.D., Reza Jafarzadeh Esfehani4,5,6 Ph.D., Saeed Abtahi1* M.D.

Background and Aims: Warfarin is an anticoagulant agent used for many years in treating various clinical conditions such as thromboembolisms in cardiovascular disease. Some patients require different doses of warfarin to reach the therapeutic international normalized ratio ratio. These patients have specific demographic characteristics. Genetic polymorphisms in specific genes have been reported to be an essential factor in response to warfarin. The present study investigated the effect of these polymorphisms of genes on warfarin dose necessities in pediatric of VCORC1 gene in patients. Material and Methods: Ninety-five patients with cardiovascular disease, who were receiving warfarin for at least three months, enrolled in the present cross-sectional study. Their genomic DNA was extracted from their peripheral blood, and the VKORC1 (rs9923231) polymorphism was evaluated by polymerase chain reaction and sequencing. Results: Among the study population, 48 patients (50.5%) had TC genotype and, 21 (22.1%) and 9 (9.5%) patients have TT and CC genotype, respectively. There was no significant relation between Warfarin dose and VCORC1 genotype in our population (p<0.05). Conclusions: The VKORC1 polymorphism (rs9923231) did not significantly affect the warfarin required for cardiovascular disease patients. Further studies evaluating other genes such as CYP2C9 polymorphisms in our population are warranted.

267-279

Authors: Hoshang Roshanmehr1* M.Sc., Morad Rostami2 M.Sc., Maryam Seyedtabib3 Ph.D., Naser Kamyari4 Ph.D.

Background and Aims: Due to the high prevalence of carbohydrate and lipid metabolism disorders worldwide and in Iran and as recommended by the international diabetes federation, about more investment in diabetes research in Iran, it is decided to perform of this study in the urban and rural area of Gotvad city in the Khuzestan Province of Iran. Materials and Methods: In this cross-sectional study, among 31822 eligible people, 4138 people aged 29 years or more participated between November 2017 and March 2018. Samples were obtained from patients, and after centrifugation and separation of serum, fasting blood sugar (FBS) and total cholesterol were measured employing the Glucose and Cholesterol Assay Kit of Pars Azmun Company. Since FBS and cholesterol statuses are ordinal, an ordinal logistic regression proportional odds model was used to identify the risk factors of responses if the proportional odds assumption satisfies. Univariate analysis was performed with independent t-test and ANOVA with a Tukey posthoc. Results: Two models were determined, and except the area in predicting cholesterol status; all other covariates, including age, FBS value, cholesterol value, sex, and area in predicting FBS, were the significant predictors of cholesterol and FBS levels. Conclusions: Although there where a high prevalence of FBS and cholesterol level disturbances, from 24.9% and 25.9% of the population with disturbances in serum cholesterol and glucose levels, respectively, nearly 80% and 25.9% of them are in the borderline high group and impaired fasting glucose states, respectively which can return them to a healthy state with appropriate and prompt interventions.

300-311

Authors: Samaneh Borooni1 Ph.D., Fahimeh Nourbakhsh2* Ph.D., Elahe Tajbakhsh3 Ph.D., Parisa Behshood4 Ph.D.

Background and Aims: The therapeutic effects of the olibanum, the resin of Boswellia serrata (B. serrata) from the Burseraceae family in inflammatory disease have been reported. There are more than 200 active ingredients in this resin, including Boswellic acid. It is proposed that aqueous extract of B. serrata can improve memory impairment induced by cerebral inflammation result in the administration of lipopolysaccharide (LPS). Materials and Methods: In this study, after the treatment of rats with LPS, brain toxicity induction was performed, and finally, the behavioral tests were evaluated. Following cerebral inflammation induction and treatment, behavioral performance biochemistry tests and molecular methods were assessed in all groups. Results: LPS administration increased the duration and distance to find the platform in the Morris water maze test compared to the control group in 5 days (p<0.05 to p<0.001). Furthermore, LPS reduced the peripheral, central, and total locomotion compared to the control group (p< 0.001) in the open field test. Pretreatment with both doses of aqueous extract of B. serrata enhanced performances of the rats in Morris water maze (p<0.05 to p<0.01) and open field test (p<0.01 to p<0.001). LPS also increased hippocampus Interleukin-6, malondialdehyde levels (p<0.001). Conclusions: Aqueous extract of B. serrata can be a useful drug in memory impairment caused by LPS-induced inflammation.

153-158

Authors: Elham Rezaee 1 M.Sc., Seyed Adnan Kashfi 2 M.Sc., Parisa Bagheri 3* M.Sc.

Background and Aims: Urinary tract infections are one of the most common human infections seen in all age groups and both sexes. Inappropriate use of antibiotics to treat urinary tract infection causes the resistance of the pathogens to the drug. The present study aimed to determine the frequency of gramnegative and gram-positive bacteria and antibiotic resistance patterns in patients with urinary tract infection. Materials and Methods: Samples were cultured on Blood Agar and Eosin Methylene Blue. Colonies' growth was identified by biochemical tests and standard microbiological and antibiotic sensitivity tests, which were performed with the disc diffusion method according to the Clinical and Laboratory Standards Institute 2016 Standard. Results and Conclusions: The isolated bacteria showed the highest susceptibility to imipenem (89.66%) and meropenem (87.21%) and the highest resistance to sulfamethoxazole (50.00%) and nalidixic acid (44.09%). So, using imipenem is recommended as the most effective antibiotic for the treatment of infection.

168-178

Authors: Amos Dangana 1 M.Sc., Anthony Uchenna Emeribe 2 M.Sc., Sanusi Musa 3 M.Sc., Lugos D. Moses 4 Ph.D., Christopher Ogar 2 M.Sc., Charles Ugwu 5 B.MLS, Chibueze Obi-George 6 B.MLS, Jessy Thomas Medugu 7 M.Sc., Olusoji Matthew Adeyemi Billyrose 1 M.Sc., Idris Nasir Abdullahi 3* M.Sc.

Background and Aims: Iron Deficiency (ID) is a common obstetric problem and nutritional disorder that occurs mostly in developing countries. Hence, nutritional studies are required every few years to determine the necessary healthcare interventions for pregnant women. This cross-sectional study evaluated the plasma iron, ferritin, transferrin, total iron-binding capacity (TIBC), and unsaturated ironbinding capacity (UIBC) levels of pregnant women referred to the University of Abuja Teaching Hospital, Gwagwalada, Abuja, Nigeria. Materials and Methods: Blood samples were collected from 58 non-pregnant women, 22 pregnant women in the first trimester, 52 in second, and 44 in the third trimester. The plasma ferritin concentration of these samples was measured by chemiluminescence assay, while the plasma transferrin, TIBC, and iron concentrations were measured by enzyme-linked immunosorbent assay. Plasma ferritin levels <15μg/L were considered ID. Results: The overall prevalence of ID in pregnant women was 33.1%. However, the prevalence of ID was 29.3%, 22.7%, 34.6%, and 36.4% among non-pregnant women, women in the first trimester, second and third trimester, respectively. The mean±SEM iron levels were significantly higher among pregnant women compared to non-pregnant women (p=0.004). There was no significant difference in the mean±SEM of plasma ferritin, transferrin and TIBC concentrations between pregnant and non-pregnant women (p>0.05). Nevertheless, the mean±SEM plasma iron, ferritin, and UIBC significantly differed across the gestational ages of pregnant women (p<0.05). Conclusions: This study revealed a high prevalence of ID during pregnancy, which increases with the trimester of affected women. Healthcare interventional measures that can address ID are recommended.

191-196

Authors: Zahra Salari 1*M.Sc., Atefeh Ranjkesh 1 M.Sc., Emad Behboudi 2 M.Sc.

Background and Aims: Helicobacter pylori (H. pylori) is a gram-negative, microaerophilic, spiral-shaped flagellated bacterium that is urease, catalase and oxidase positive. One of its pathogenicity factors is the iceA gene. H. pylori has recently been recognized as a genetic indicator for the development and evolution of duodenal ulcer disease in the East. This study aimed to determine the presence of this bacterium in gingival plaques in non-endocrine patients in Bojnourd city, and the polymerase chain reaction technique examined the percentage of iceA gene. Materials and Methods: A total of 100 samples of dental plaque were taken and transferred to a tube that has been physiologically placed. After DNA extraction, primer design was performed, and then the polymerase chain reaction was performed for the whole sample. Results: Of 100 samples examined in this study, two samples of H. pylori were positive (2%), and the frequency of the iceA gene of two samples was positive (100%). Conclusions: In the Bojnord city, the frequency of iceA gene in people is high, and the frequency of H. pylori in tooth plaques is low. Also, iceA gene can be considered as an indicator for predicting the contamination and risk of H. pylori infection in the region. To confirm the results, more molecular studies are required in other populations.

212-223

Authors: Ogechukwu F Odo 1,2* Ph.D., Nkiruka C Azubuike 2 Ph.D., Peter U Achukwu 2 Ph.D., Michael C Ugwu 3 M.Sc., Chisom H Udeogu 3 M.Sc., Ozoemena C Ike 4 M.Sc.

Background and Aims: Cola acuminata pod extract (CAPE) is used in Nigerian traditional medicine to treat low sperm count in males. Hence, this study aimed to investigate the effects of CAPE on the reproductive system of male albino rats. Materials and Methods: Preliminary acute toxicity testing, phytochemical screening, proximate analysis, and determination of vitamin E concentration were performed on the extract. Three groups (I, II, and III) of 12 rats were used in the study. Each group comprised three batches (A, B, and C) of 4 rats each for different periods (4, 6, and 8 weeks, respectively). Results: Acute toxicity testing showed that CAPE had an oral lethal dose of 50% (LD50) of >5 g/kg body weight (b.wt) in rats. Vitamin E concentration was 0.511 mg/l. The caudal epididymal sperm count increased marginally from 4 to 6 weeks for rats treated with 800 mg/kg b.wt., but reduced significantly at 8 weeks for both CAPE treatment groups (II and III) (p<0.05) compared with the control group (I). Significant reduction (p<0.05) of sperm motility and serum testosterone levels at 8 weeks were observed. The histology of the reproductive organs revealed evident histo-architectural changes. Conclusions: These results suggest that the aqueous pod extract of Cola acuminata causes marked alterations in reproductive organs and shows antispermatogenic and antiandrogenic effects when administered orally over 8 weeks in mature male rats leading to contradicting its use as a traditional remedy for low sperm count in males.

67-71

Authors: Asma Kheirollahi , Azam Karkhaneh *

Background and Aims: Non-high density lipoprotein cholesterol (non-HDL-C), which reflects all cholesterol present in potentially atherogenic lipoprotein particles, might be a useful marker of atherosclerosis in diabetic subjects. In the present study, we evaluated the prevalence of high non-HDL-C in patients with dyslipidemia in diabetic and non-diabetic subjects following LDL-C assessment as the first goal of therapy. Materials and Methods: A data set of 2142 individuals was included in the study. All values of lipid profile were compared between non-diabetic and diabetic groups and the prevalence of dyslipidemia was evaluated in two groups. Results and Conclusions: According to the results, 48% of patients with diabetes achieved combined LDL-C ≤2.5 mmol/L and non-HDL ≤3.3 mmol/L targets, and 58.2% of diabetic patients achieved LDL-C goal while only 50.2% attained non-HDL-C goal. Also, the results indicated that non-HDL-C significantly heightened in patients with diabetes. Therefore, non-HDL-C needs to be calculated as a routine assessment in patients with diabetes.

90-101

Authors: Hamid Reza Jamshidi * , Hasti Kalantar

Background and Aims: Mercury, with its oxidative activity, causes damage to the antioxidant enzymes thus resulting in physiological disorders. Sodium selenide is an antioxidant that protects antioxidant enzymes. The aim of this study was to investigate the protective effect of sodium selenide on renal toxicity induced by mercuric chloride in rats. Materials and Methods: Animals were divided into 6 groups of 6 each. Control group was treated with mercuric chloride (2.5 mg/kg) subcutaneously. Groups 2, 3 and 4 of the rats were treated with 2.5 mg/kg of mercuric chloride and sodium selenide subcutaneously administered at doses of 0.02, 0.04 and 0.08 mg/kg, respectively. The fifth group received 0.04 mg/kg sodium selenide subcutaneously after 25 days of mercuric chloride (2.5 mg/kg) treatment. The sixth group was treated with 2.5 mg/kg of mercuric chloride subcutaneously and 100 mg/kg of vitamin E by gavage. Results: After data analysis of the activity of the enzyme catalase, superoxide dismutase, glutathione peroxide, creatinine and urea, there was a significant difference in the activity of these factors compared with the control group in blood and kidney tissue (p<0.05). Conclusions: This study indicated that mercuric chloride has toxic effects in blood and kidney and sodium selenide may be able to reduce its’ blood and renal toxicity.

110-120

Authors: Sakineh Jam Shahriari, Fahimeh Alizadeh * , Alireza Khodavandi

Background and Aims: Candida albicans (C. albicans) is the most common opportunistic human pathogen. Therapeutic options for Candida infections are limited to available antifungal drugs. The aim of this study was to investigate the effects of fluconazole/clotrimazole (FLU/CLT) on C. albicans hyphae formation. Materials and Methods: We have established the effectiveness of the combination of FLU/CLT on C. albicans hyphae formation. Interaction of C. albicans with combination of FLU/CLT was performed using the CLSI guidelines and time-killing curves. We investigated the anti-hyphal activities of combination of FLU/CLT against C. albicans using XTT and crystal violet assays as well as scanning electron microscopy and expression of HWP1 gene. Results: The interaction of C. albicans with FLU/CLT resulted in synergistic, partial synergistic and indifferent effects. The interaction of FLU/CLT were confirmed by time-killing curves. FLU/CLT combined resulted in the reduction of metabolic activity and hyphae formation in C. albicans. Images taken by scanning electron microscopy indicated the effectiveness on hyphae disruption. According to relative real time polymerase chain reaction analysis, the mean Ct values revealed the significant decrease in expression level of the HWP1 gene. A 2.86- and 2.33-fold decrease in HWP1 gene expression was observed in combination of FLU/CLT treatment at 2× minimum inhibitory concentration and 1× minimum inhibitory concentration, respectively (p=0.002). Conclusions: We confirmed that the hyphae is a target for the combination of FLU/CLT in C. albicans. HWP1 gene is likely to be considered as a probable targets synergistic interaction of FLU/CLT against C. albicans.

128-137

Authors: Zeinolabedin Sharifian, Batool Hashemibeni , Majid Pourentezari * , Ali Valiani , Mohammad Mardani , Mohammad Zamani Rarani , Hamidreza Pourentezari

Background and Aims: Tissue engineering is a relatively novel field that has been intensely developing during recent years and has shown to be excessively promising when used for cartilage regeneration. Scaffolds represent important components for tissue engineering. Materials and Methods: The Poly Lactic-Co-Glycolic Acid (PLGA) impregnated with fibrin and hyaluronic acid (HA) produce hybrid scaffolds. human adipose-derived stem cells (hADSCs) were seeded in scaffolds and cultured in chondrogenic media. The viability of cells in different groups was assessed by MTT. The expression of chondrogenic related genes [Sox9, type II collagen (Col II), Aggrecan(AGG)] and type X collagen (Col X) was quantified by real-time polymerase chain reaction. Results: The results of the real-time PCR showed SOX9, AGG and Col X gene expression in the control groups being significantly lower than the other groups (p≤0.05). It also demonstrated Col II gene expression in the control group being significantly lower than the PLGA/Fibrin and PLGA/Fibrin/HA groups (p≤0.05). The Col X gene expression of cells in PLGA/HA and PLGA/Fibrin/HA groups significantly decreased in comparison with the PLGA/Fibrin group (p≤0.05). Conclusions: These conclusions indicate that administration of PLGA/ Fibrin and PLGA/HA scaffolds, particularly PLGA/Fibrin/ HA, motivates chondrogenesis in hADSCs. This can be diminished by decreasing hypertrophic markers and increasing characteristic markers of hyaline cartilage.

1-8

Authors: Nasrin Hadi, Faezeh Namazi, Fatemeh Ketabchi Farinaz Khosravian, Parisa Ravaghi, Mansoor Salehi

Background and Aims: Multiple sclerosis (MS) has been assumed to be a complex and indecipherable disease, and poorly understood with regard to etiology which is characterized by relapses and remissions. The expression of microRNAs (miRNAs) is known to be associated with the regulation of immune responses. Recently, investigations have reported that miRNA expression profiles in blood cells become changed in MS. The aim of this study was to elucidate the alterations in the expression of circulating miR-377 and miR-98 in 60 relapsing-remitting MS (RRMS) patients in comparison with controls. Materials and Methods: This study was conducted using a quantitative real-time polymerase chain reaction method to explore the expression of circulating miR-377 and miR-98 in peripheral blood mononuclear cells from 60 RRMS patients, 30 of whom were recurring patients, 30 were two months after relapse patients, and 30 others were controls, in order to examine the association of expression level of these miRNAs with RRMS. Results: Results indicated that the expression of miR-377 significantly increases in recurring patients and two months after relapse patients in comparison with controls (p=0.0017 and p=0.0001, respectively). However, miR-98 demonstrated down regulation in recurring patients and two months after relapse patients (p=0.0002 and p=0.0001, respectively). Conclusions: It can be concluded that miR-377 and miR-98 may be prospective biomarkers with the potential use for diagnosis of RRMS patients in the future investigations.

15-22

Authors: Hamideh Valizadeh, Gilda Eslami, Reyhane Rahnama, Abolghasem Rahimdel, Fateme Sadat Dashti, Reza Mansouri

Background and Aims: Multiple sclerosis (MS) is known as a partially inheritable inflammatory autoimmune disease which involves the nervous system. Different studies suggest that immune dysregulation has an important role in the pathogenesis of MS, but its exact pathomechanism has not yet been explicated. The receptor for advanced glycation end products (RAGE) is a member of the immunoglobulin superfamily of cell surface molecules. In MS patients, the expression of membrane-bound RAGE on peripheral blood mononuclear cells, as well as the serum levels of the RAGE ligands are upregulated. Materials and Methods: In this case-control study, we evaluated the association between MS incidence and RAGE p.82G>S polymorphism (rs2070600) compared with healthy controls in an Iranian population. A total of 158 patients and 156 healthy blood donors were enrolled. This polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism. Results: Allele frequencies (p=0.319) and genotype distribution (p=0.315) of the RAGE p.82G>S gene polymorphism in MS patients and controls were not significantly different. Conclusions: The present study indicated no relationship between RAGE p.82G>S polymorphism and the risk of MS in Iranian population.

30-34

Authors: Danyal Daneshdoust , Durdi Qujeq , Mohesn Vakili Sadeghi , Ali Mahmoudi, Hamed Ghasmtabar

Background and Aims: There is now an increasing body of evidence that survivin is a protein, expressed highly in breast cancer. The signaling interaction of protein survivin in breast cancer is still unclear, but physiological regulation of survivin seems to be linked to the breast cancer occurrence and severity of it. Materials and Methods: Serum samples were obtained from April 2015 to November 2017 out of women enrolled in a group undergoing annual breast cancer testing. Routine blood samples were analyzed at the Biochemical Laboratory of Rouhani Babol University Hospital . Results: Serum levels of survivin in patients with breast cancer group increased compared to the healthy controls [207.520±110.284 (mean±SD) vs. 126.212±53.130, ng/L, p<0.001]. Also, we detected a positive correlation between elevated serum survivin level and clinical characteristics of patients with breast cancer. Conclusions: Serum survivin measurement was shown to discriminate patients with breast cancer from healthy controls. However, further studies are needed to confirm this role and its benefits.

41-48

Authors: Khodayar Ghorban, Maryam Dadmanesh, Mahmood Sheikh Fathollahi, Zeynab Rezai, Paria Ezati, Mohammad Hossein Salmani, Fatemeh Akrami Mohajeri, Hasan Yousefi-Daredor,Hasan Ebrahimi,Derek Kennedy, Mohammad Kazemi Arababadi

Background and Aims: The critical aim of this study was to evaluate the effects of breast-milk contamination with Nickel and Arsenic, as well as aflatoxin M1 (AFM1) on the milk levels of IL-6 and IL-8. Materials and Methods: Breast milk was collected from 76 mothers on the 30th day post-parturition and milk levels of AFM1, Nickel, Arsenic, IL-6 and IL-8 were evaluated. Results: The results showed 8 and 29 out of 76 collected samples being contaminated with AFM1, Nickel and Arsenic, respectively. Levels of IL-6 and IL-8 in the milk samples did not differ in AFM1 and Nickel and Arsenic contaminated milk compared with the normal controls (non-contaminated milks). There was a moderate positive relation between milk levels of IL-8 and Arsenic. Conclusions: Although AFM1, Nickel and Arsenic are the inducers of cytokine production; they are unable to induce secretion of IL-6 in breast milk. Arsenic can be a trigger of IL-8, which is a strong chemoattract for neutrophils. Thus, it appears that Arsenic may induce an inflammation in milk in IL-8 dependent manner.