Tehran University Medical Journal

A glance into the future cardiac stem cells: review article

Ali Mohammad Alizadeh

It was assumed that the loss of cardiomyocytes is irreversible. The main goal is to develop widely available and clinically applicable treatments for heart diseases. The several studies have showed that the use of stem cells can improve complicacies such as cardiovascular diseases. Stem cells have a potential benefit of the self-renewal and cell differentiation into the cell types that can play an important role in the organogenesis and the embryonic development. In a lifetime, the heart muscle has a population of cardiac stem cells (CSCs) in which a dramatically increase after cardiovascular damages. So far, seven types of CSCs have been discovered with the different molecular phenotype and the cell differentiation potential. In this regard, the proliferation and the differentiation increase of CSCs in the cardiac ischemic areas can be a key factor to improve heart complicacies. Paracrine and/or autocrine factors, the extracellular matrix and the genetic mediators including microRNA can control the function of CSCs. It has clearly been understood that the factors mentioned previously have the ability to improve these complicacies. The differentiation, the survival and the self-renewal of CSCs are largely under the control of factors in the heart microenvironment. Several studies showed that the cytokines and the growth factors play the important role in the proliferation and the migration of CSCs. Taking advantage of these factors together CSCs to repair damaged heart can enhance this method efficiency. This review will discuss the different kinds of CSCs, their molecular phenotype and cardiac regeneration potential in order to improve cardiovascular diseases. It seems that CSCs-based therapy is emerging as a novel approach for myocardial repair over conventional cardiovascular therapies. Therefore, understanding the new aspects on the molecular mechanisms and the signaling pathways involving CSCs is critical for the development of the therapeutic strategies in cardiac patients that would be valuable for researchers in both fields of molecular and clinical cardiology.

The effect of BLA GABAB receptors in anxiolytic-like effect and aversive memory deficit induced by ACPA

Mohamreza Zarindast

Background: As a psychoactive plant, Cannabis sativa (Marijuana) is widely used throughout the world. Several investigations have indicated that administration of Marijuana affects various cognitive and non-cognitive behaviors. These include anxiety-like behaviors and learning and memory deficit. It has been shown that three main cannabinoid receptors [i.e. CB1, CB2 and CB3 are involved in cannabinoids’ functions. CB1 receptors are abundantly expressed in the central nervous system regions such as hippocampus, amygdala, cerebellum and the cortex. Therefore, the neuropsychological functions of endocannabinoids are thought to be more linked to CB1 receptors. Among other brain regions, CB1 is highly expressed in the amygdala which is an integral component of the limbic circuitry. The amygdala plays a major role in the control of emotional behavior, including conditioned fear and anxiety. In present study we examined the possible roles of basolateral amygdala (BLA) GABA_B receptors in arachydonilcyclopropylamide (ACPA)-induced anxiolytic-like effect and aversive memory deficit in adult male mice.

Methods: This experimental study was conducted from September 2013 to December 2014 in Institute for Studies in Theoretical Physics and Mathematics, School of Cognitive Sciences, Tehran and Male albino NMRI mice (Pasture Institute, Iran), weighting 27-30 g, were used. Bilateral guide-cannulae were implanted to allow intra BLA microinjection of the drugs. We used Elevated Plus Maze (EPM) to examine memory and anxiety behavior (test-retest protocol). ACPA administrate intra-peritoneal and GABAB agonist and antagonist administrated intra-amygdala.

Results: Data showed that pre-test treatment with ACPA induced anxiolytic-like and aversive memory deficit The results revealed that pre-test intra-BLA infusion of baclofen (GABA_B receptor agonist) impaired the aversive memory while phaclofen (GABA_B receptor antagonist) improved it. Interestingly, pretreatment with a sub-threshold dose of baclofen reversed and potentiated anxiolytic-like effect and aversive memory deficit induced by ACPA, respectively. Conversely, similar effect with sub-threshold dose of phaclofen showed that this drug only restored aversive memory deficit but did not alter anxiolytic-like effect induced by ACPA.

Conclusion: Data indicated that BLA GABA_B receptors have critical and different roles in anxiolytic-like effect and aversive memory deficit induced by ACPA.

Evaluation of fluconazole effect in prevention of fungal infection and mortality and morbidity in very low-birth-weight infants

Samileh Noorbakhsh

Background: Fungal infections especially Candida species are frequent cause of mortality and morbidity in very low-birth-weight (VLBW) infants receiving intensive care; Candida infections are tissue invasive. This infection increases the risks of adverse neurodevelopmental sequelae. Prevention and treatment of fungal infection is so important in very VLBW infants. The aim of this study was to determine the prophylactic effect of fluconazole in decreasing the mortality and morbidity in VLBW infants (less than 1500gr) admitted in NICU.

Methods: This prospective case control study were conducted among 102 Infants (weighing less than 1500gr at birth at born) admitted in NICU department of Ali Asghar University Hospital from 2012 to 2013, Tehran, Iran. Weigh of birth in cases and groups were less than 1500 gr and both were culture negative. Cases received oral fluconazole 3 mg/kg in 3 days in 1^st and 2^nd weeks, alternate day in 3^rd and 4^th weeks, daily in 5^th and 6^th weeks. Control groups had not received fluconazole. Mortality and morbidity and hospital stay were compared between cases and controls groups.

Results: We studied 49 very low-birth-weight infants with negative culture as cases (received fluconazole prophylaxis), 46 VLBW infants without fluconazole profilaxy (controls). No significant difference in gestational age (P=0.2), and mean weights (P=0.4) were observed between cases and controls. The mortality rate 8.7% (n=4) in controls (without prophylactic fluconazole) observed vs 2% (n=1) mortality rate in VLBW cases (with prophylactic fluconazole). Although the mortality rate in controls was 4 times higher than cases, but without significant differences (P=0.1). Indeed, mean duration of hospital stay in controls was longer than cases (28.41±9.93 vs 19.85±6.19 days, P=0.00001).

Conclusion: Although prophylactic fluconazole in VLBW could decrease the mortality of cases (control the fungal infection) 4 fold in compare with controls (no treatment), it was not significant. The prophylactic effect of fluconazole might decrease the length of hospital stay of VLBW neonates in NICU. Due to limited number of cases and control. For further decision about prophylactic use of fluconazole, prospective RCT studies with larger cases and control would be helpful in future.

Urinary incontinence and pelvic girdle pain during pregnancy: a clinical trial

Mahboobeh Shirazi

Background: To examine the association among pelvic girdle pain (PGP), urinary incontinence (UI) and pelvic floor muscle (PFM) function in pregnant women in second and third trimester.

Methods: 300 pregnant women who admitted for standard pregnancy care were enrolled in the study. Urinary incontinency was measured via the international consultation on incontinence questionnaire short form. Pelvic girdle pain was diagnosed according to existing guidelines. Vaginal examination assessed pelvic muscles contract- relax patterns and muscle strength. The software stata version 13 (Stata Corp., TX, USA) was used for data analysis.

Results: Overall 300 women (150 with PGP and 150 without PGP) were included in final analyses. There was not significant differences between the demographic data including, body mass index before pregnancy, maternal age, mode of delivery. Prevalence of urinary incontinence in women with pelvic girdle pain was 41.5 percent (CI 95%: 32.01- 51.48) while the prevalence of urinary incontinence in women without pain was 21.9 percent (CI 95%: 14.99-30.03). Using logistic regression, the relationship between urinary incontinence and pelvic girdle pain was significant. (CI 95%: 1.07-3.31, P=0.02). Multivariate logistic regression analysis was used to evaluate the relationship between PGP and pelvic floor muscle function and results showed that pelvic floor muscle strength in women with PGP was significantly lower than women without PGP. (CI 95%: 0.24-0.68, OR= 0.4, P<0.001). The duration of the contraction of the pelvic floor muscles in patients with PGP was significantly shorter than women without PGP. (CI 95%: 0.21-0.60, OR =0.35, P<0.001).

Conclusion: There was a significant association between pelvic girdle pain and urinary incontinence during the second and third trimesters of pregnancy and also the pelvic floor muscle strength and duration of the contraction of the pelvic floor muscles in women with pelvic girdle pain was significantly lower than those without PGP.

Investigation of hTERT gene expression levels in two cell lines infected by high-risk human papilloma virus

Mahdi Kamali

Background: Human papilloma virus (HPV) is one of the most important factors in cervical cancer. Viral sequences are integrated into the host cell genome. In mild cases the virus causes skin damages, in severe cases it leads to cancer. Like many other cancers, telomerase gene expression was increased in cervical cancer. This enzyme is a reverse transcriptase that contains two common subunits: i) catalytic protein called human telomerase reverse transcriptase (hTERT) and, ii) RNA sequence called hTR. hTERT expression is hardly found in any somatic tissues. Detection of high telomerase activity in human cells, lead to tumor genesis. So hTERT can be used as a diagnostic tool in cancer detection.

Methods: This experimental study was carried out from May 2013 to April 2014 in Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences in Tehran, Iran. Caski and Hela cancer cell lines were used which contain HPV16 and HPV18 respectively. Cell lines were cultured and total RNA was extracted. Following normalization agent glyceraldehyde-3-phosphate dehydrogenase (GADPH), hTERT expression level was determining by real-time PCR method. For each sample, the expression level of hTERT and GAPDH were quantified as copy numbers (per reaction) using the standard curve. Finally, hTERT levels in Hela and Caski cell lines were compared quantitatively by t-test using GraphPad statistic software version 5 (San Diego, CA, USA).

Results: According to the charts real-time PCR, hTERT gene expression in Hela and Caski cancer cell lines is significantly different (t=0.0319).

Conclusion: All results confirm that hTERT expression levels in Hela and Caski cell lines are significantly different and the level of hTERT expression in the Caski cell line was slightly higher than that of Hela cell line. The significant difference between hTERT mRNA expression levels reported here could be used as a tumor marker for HPV16 and HPV18 in cervical cancer.

Comparison of serum vitamin D levels in women with gestational trophoblastic neoplasia and healthy women

Malihe Hasanzadeh

Background: Gestational trophoblastic neoplasia (GTN) consists of a broad spectrum of benign and malignant tumors which are stem in human placental trophoblast. Vitamin D has several biologic rules. Among the effects of vitamin D on cells, we could mention induction of differentiation and apoptosis and also inhibition of proliferation, angiogenesis and metastatic potency. As nutrition plays a pivotal rule in GTN, in this study we compared serum 25-hydroxy-vitamin D [25(OH) vitamin D] in patients with GTN and normal pregnant women.

Methods: In this prospective case-control study, 30 GTN patients and 48 normal pregnant women were considered as control group who referred to Qaem University Hospital, Mashhad, Iran, from July 2013 to October 2015 were included. All included persons to the study had no history of chemotherapy or using vitamin D supplements. After filling informed consent forms and recording demographic data, 25(OH) vitamin D serum level were measured in both group by enzyme-linked immunosorbent assay (ELISA) method.

Results: Age in two groups was the same (P=0.565). There was no significant difference in gestational age between two groups (P=0.887). Pathologic diagnosis in 83.33% (25 patients) was complete hydatidiform mole and in 16.67% (5 patients) was partial hydatidiform mole. 25(OH) vitamin D serum level in 73.3% of GTN patients and 2.1% of normal pregnant women was lower than 10 ng/ml and among all participants, only 6.3% of pregnant patients had 25(OH) vitamin D serum level higher than 30 ng/ml. Mean serum level of 25(OH) vitamin D in GTN patients was significantly lower than pregnant women group (9.09±5.46 vs. 20.06±6.37, P=0.000). 25(OH) vitamin D serum level between complete and partial hydatidiform mole groups had no significant difference (P=0.384).

Conclusion: Altogether, it was observed that 25(OH) vitamin D serum level is significantly lower in women with GTN than normal pregnant women. Modifying serum levels of vitamin D in molar pregnancy with low level of vitamin D may prevent the development of GTN.

Association of vitamin D levels and receptor gene polymorphisms with medullary thyroid cancer

Mohammad Mazani

Background: Thyroid cancer is the most common endocrine malignancy. Accounting for approximately 1-2% of all cancers. Thyroid cancers have been divided into four main types: papillary, follicular, medullary and anaplastic. The active form of vitamin D (1,25- (OH) 2-vitamin D3) by binding to its receptor, using genomic and non-genomic mechanisms inhibits the proliferative effect of TSH on thyroid cells. Therefore, vitamin D may have a role in regulating of thyroid gland cell proliferation. Many studies have shown anti-cancer effects of vitamin D in cancers. Polymorphisms of Vitamin D receptor can influence the prevalence to various cancers. In the present study, serum level of vitamin D and FokI, BsmI and Tru9I polymorphism of vitamin D receptor was investigated.

Methods: This case-control study was performed in the summer of 2015 in Endocrinology and Metabolism Center of Shahid Beheshti University of Medical Sciences, Tehran, Iran. Forty medullary thyroid cancer patients and 40 healthy controls were investigated. Genomic DNA of subjects was extracted with saturated salt/proteinase K and polymorphisms of vitamin D receptor gene investigated by polymerase chain reaction-sequencing. Serum level of vitamin D evaluated by ELISA technique. The results were analyzed by SPSS, ver. 20 (Chicago, IL, USA) and GraphPad Prism, ver. 5 (GraphPad, Inc., CA, USA) softwares.

Results: Genotypic and allelic abundance of FokI and BsmI polymorphisms between test and control groups have not shown significant different. In Tru9I polymorphism, Tt genotype abundance in test group were 45 percent and in control group were 17.5 percent and t allelic abundance in test group were 25 percent and in control group were 8.7 percent which this different were significant. Average serum level of vitamin D in test group was 23.32 ng/ml and in control group was 18.95 ng/ml which was statistically significant.

Conclusion: Unexpectedly, serum levels of vitamin D in test group were higher than control group. Tru9I polymorphism is significantly correlated to medullary thyroid carcinoma prevalence.

Comparison of the expression levels of Fas and Apaf-1 genes in systemic sclerosis dermal fibroblasts

Majid Abed Khojasteh

Background: Systemic sclerosis (SSc) is an autoimmune rheumatic connective tissue disease. In normal wound healing process, fibroblasts are activated, proliferated and involved in tissue repair, and then removed by apoptosis. In systemic sclerosis, patient’s fibrosis occurs when fibroblasts become resistant to apoptosis and secrete a large amount of collagen and other extracellular matrixes. As the primary causes the disease are very complex and often unknown, it is necessary to consider or target the secondary causes of disease, such as the unresponsiveness of activated fibroblasts to apoptosis as the major factor in the creation and deployment of illness. In this study, we examined the expression levels of two key pro-apoptotic genes, Fas and Apaf-1, which are respectively involved in external and internal pathway of apoptosis.

Methods: In a case-control study skin biopsy samples were obtained from 19 patients with diffuse SSc, and 16 healthy controls. Dermal fibroblasts were cultured and total RNA was isolated from cell populations using High Pure RNA Isolation Kit (Roche Applied Science, Mannheim, Germany), followed by cDNA synthesis using RevertAid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific Inc., Massachusetts, USA). Real-time PCR was performed using SYBRGreen gene expression master mix (Takara Shuzo, Co., Ltd, Shiga, Japan) and specific primers for Fas and Apaf-1. Real-time data were analyzed using the (2^-ΔCT)×1000 method. Statistical analysis was accomplished by using the SPSS software, v22 (IBM, Armonk, NY, USA). The P value less than 0.05 were recognized as a significant threshold. All data are represented as the mean ± SEM.

Results: Our results showed no significant difference in Fas (P=0.8) and Apaf-1 (P=0.17) mRNA expression levels between skin fibroblasts of systemic sclerosis patients and healthy controls.

Conclusion: In this study we observed no significant change in Apaf-1 and Fas mRNA levels in systemic sclerosis fibroblasts compared to control group. Hence, Apaf-1 and Fas are not transcriptionally activated in SSc fibroblasts. Further studies need to take place on protein levels and function of these proteins to confirm the mRNA transcription results.

High-sensitive C-reactive protein in patients with acute coronary syndrome in statin therapy and its impact on prognosis

Hamed Tavan

Background: Since high plasma level for C-reactive protein (CRP) is a risk factor for cardiovascular disease, thereby decrease in the level of high- sensitivity C-reactive protein (hs-CRP) in acute coronary syndrome (ACS) patients through anti-inflammatory drugs can reduce mortality and the incidence of heart failure. Accordingly, this research aims to investigate the effect of hs-CRP on ACS patients before and after treatment with astatines.

Methods: This cross-sectional and cohort study was performed for the population of 90 patients with acute coronary syndrome (ACS) martyrs at the Mustafa Khomeini University Hospital in the Ilam city, Iran, From July to September, 2014. Blood samples were collected at admission and demographic and clinical symptoms, echocardiography and electrocardiography were recorded. At admission, the questionnaire including demographic information and medical history of patients was filled by the researchers and echocardiography and physical examination was carried out by cardiologist. The obtained data are further explored and analyzed via SPSS software, ver. 19 (Chicago, IL, USA).

Results: The sample under study was 52.2% and 48.8% men and women, respectively. Phi correlation coefficient of 73% and positive Cramer's V of 0.879 was observed between re-admission and arrhythmia admission for the group received 40 mg atorvastatin. It means that we have more re-admission when arrhythmia increases. Only 4% correlation coefficient and very low positive Cramer's V of 0.293 was seen for the group who receive 80 mg atorvastatin. It indicates that no significant correlation exists between eject fraction of admission and re-admission (P=0.18). The results showed that hs-CRP of the group that received 80 mg atorvastatin was 0.179 which is lower than 0.37 for the group who received 40 mg atorvastatin.

Conclusion: By increasing the astatine dose, the amount of hs-CRP and consequently the risk of subsequent cardiovascular events were reduced. Hence, high starting dose of atorvastatin at preliminary stages of hospitalizing can reduce re-admission and cardiovascular consequents.

Fetal cardiac rhabdomyoma: case report

Fakhrosadat Sajjadian

Background: The primary manifestation of cardiac tumors in embryonic period is a very rare condition. Cardiac rhabdomyomas most frequently arise in the ventricular myocardium, they may also occur in the atria and the epicardial surface. In spite of its benign nature, the critical location of the tumor inside the heart can lead to lethal arrhythmias and chamber obstruction. Multiple rhabdomyomas are strongly associated with tuberous sclerosis which is associated with mental retardation and epilepsy of variable severity. Ultrasonography as a part of routine prenatal screening, is the best method for the diagnosis of cardiac rhabdomyomas. In the review of articles published in Iran, fetal cardiac rhabdomyoma was not reported.

Case presentation: We report a case of cardiac rhabdomyoma on a 24-year-old gravid 1, referred to Day Medical Imaging Center for routine evaluation of fetal abnormalities at 31 weeks of her gestational age. Ultrasonographic examination displayed a homogenous echogenic mass (13×9mm), originating from the left ventricle of the fetal heart. It was a normal pregnancy without any specific complications. Other organs of the fetus were found normal and no cardiac abnormalities were appeared. No Pericardial fluid effusion was found. The parents did not have consanguineous marriage. They did not also have any specific disease such as tuberous sclerosis.

Conclusion: The clinical features of cardiac rhabdomyomas vary widely, depending on the location, size, and number of tumors in the heart. Although cardiac rhabdomyoma is a benign tumor in many affected fetuses, an early prenatal diagnosis of the tumor is of great significance in making efficient planning and providing adequate follow up visits of the patients and the complications such as, heart failure and outlet obstruction of cardiac chambers.

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Masoud Mohammadi

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