فهرست مطالب

  • Volume:19 Issue: 1, 2020
  • تاریخ انتشار: 1398/11/12
  • تعداد عناوین: 31
  • Farzaneh Dastan, Payam Tabarsi*, Majid Marjani, Afshin Moniri, Seyed MohammadReza Hashemian, Maria Tavakoli Ardakani, Ali Saffaei Pages 1-2
  • Ali PARLAR *, Seyfullah ARSLAN Pages 3-17
    Asthma is a disease characterized by spontaneous contraction of the airways in response to a wide variety of endogenous and exogenous stimuli. Many asthma models are used to mimic the human asthma model in the literature. In order to better understand the role of the cannabinoid (CB) 2 receptor in the ovalbumin (OVA)-induced asthma model, a combination of both selective CB2 agonist (AM1241) and antagonist (AM630) was used to improve inflammatory hypersensitivity and edema in rats. In the present study, it was found that OVA decreased body weight (p<0.05), increased lung weights (p<0.05), increased diastolic and systolic blood pressure (p<0.001), and caused to irregularity in pulmonary functions (p<0.001). Moreover, CB2 agonist was found not to reduce body weight, cause blood pressure and respiratory irregularities (p< 0.05). OVA led to increase in IgE, TNF-α, IL-4, MDA level (p<0.001) and total WBC count (p<0.05). CB2 treatment caused to reduce the number of total WBC and the level of total protein in BALF, to hinder to increase level of MDA, IgE, TNF-α and IL-4 (p<0.05) in BALF or serum or lung tissue. But CB2-antagonist treatment prevented the protective effect of CB2 agonist. The aim of this study was to study the role of the CB2 receptor in the OVA induced asthma model, for improve inflammatory hypersensitivity, and edema in rats. The results suggested that CB2 agonist administration to OVA induced asthmatic rats via anti-asthmatic potential through inhibition of parameters such as IgE, IL-4, TNF-α, microvascular escape, and oxidative stress.
    Keywords: CB2 receptor, Ovalbumin, Asthma, CB2 agonist, AM1241
  • Emrah Dural*, Nilay Sedes Baskak, Hatice Özcan, Yağmur Kırd, Bora Başkak, Halil Sinan Süzen Pages 18-30

    Determination of mirtazapine during psychopharmacotherapy in biological fluids is essential to achieve successful therapy, to avoid toxicity related to drug interactions, genetic variability and poor compliance. A new, rapid and sensitive high-performance liquid chromatography method has been developed in human plasma for the determination of mirtazapine (MRP) and N-desmethylmirtazapine (NDM) that is an active metabolite. The separation was achieved on a reverse phase C18 250 x 4.6 mm i.d., ODS-3 column at 40°C, using a programmed gradient elution. 20 mM potassium phosphate buffer (pH 3.9), acetonitrile and triethylamine (75.0:24.9:0.1, v/v/v) were used as mobile phase A. Mobile phase B consisted of absolute acetonitrile. Clozapine was used as an internal standard. The method showed linearity with good determination coefficients (r2>0.998) for each analyte. Intra-day and interday assay precisions (RSD%) were found less than 3.4 and 2.9 for MRP and NDM respectively. The intra-day and interday accuracy (RE%) of the method were calculated between (-2.8) and 3.5. A new extraction method was used in the study and an excellent efficiency values for MRP and NDM (94.4%, 106.6%; respectively) was obtained. The method was specific and sensitive as the limits of detection were 0.17 for MRP and 0.15 ng/mL for NDM. This method was applied to patients who received MRZ (n=62) at the 15-30 mg/day. The obtained and statistically evaluated plasma MRP and NDM levels which were 28.6±13.8 and 12.3±6.5 (mean±SD). The described procedure is relatively simple, precise, and applicable for routine therapeutic drug monitoring, especially in psychiatry clinics and toxicology reference laboratories.

    Keywords: Mirtazapine, N-desmethylmirtazapine, Plasma, HPLC-UV, Therapeutic Drug Monitoring, Validation
  • Hamidreza Jamaati, Farzaneh Dastan *, Payam Tabarsi, Majid Marjani, Ali Saffaei, Seyed Mohammadreza Hashemian Pages 31-36
    COVID-19 is currently causing concern in the medical community as the virus is spreadingaround the world. It has a heavy global burden, particularly in low-income countries. The clinicalspectrum of COVID-19 pneumonia ranges from mild to critically ill cases and Acute RespiratoryDistress Syndrome. An expert panel was held and an internal protocol was developed to managethe COVID-19 induced ARDS according to WHO recommendations and NIH guidelines.Different therapeutic regimens were employed on this protocol based on the ARDS severity andthe patients’ special characteristics. The mortality rate, the rate of survivors, and non-survivorswere reported. Of the 231 suspected cases of COVID-19 admitted to the hospital during twoweeks, 72 patients were admitted to ICU with diagnosis confirmed by RT-PCR. In total, mortalityin the ICU was 25% (n = 18) among ARDS patients over two weeks. COVID-19 induced ARDSis a major concern. The rapid progression of ARDS needs specific protocol based on patients’characteristics and rapid action.
    Keywords: COVID-19, Coronavirus, Acute respiratory distress syndrome, Treatment
  • Mahdiyeh Moudi, Ramin Saravani *, Saman Sargazi Pages 37-45
    One of the most important modulators involved in controlling apoptosis induction and viability of cancerous cells is nucleostemin. Some studies revealed that NS is also needed to maintain the proliferation of embryonic neural stem cells and early embryogenesis. This study was designed to better elucidate the association between NS depletion status and apoptosis induction of both MCF-7 and MDA-MB-468 cell lines. We examined the effects of nucleostemin-targeting siRNA on the expression of NS in MCF-7 and MDA-MB-468 human breast cancer cell lines by the Real Time polymerase chain reaction method. In addition, we investigated the correlation between knockdown of NS and viability rates and apoptosis induction in MCF-7 and MDA-MB-468 cell lines using the MTT assay and flow-cytometry technique, respectively. The NS-targeting siRNA inhibited the viability of cells in a dose- and time-dependent manner and induced apoptosis after 48 h in the cells. Thus, consistent with previous articles, this protein can be one of the regulators related to the inhibition of apoptosis and the increased viability of tumor-initiating cells in human breast cancer cell lines as well as other cancers.
    Keywords: nucleostemin, MCF-7, MDA-MB-468, siRNA, Apoptosis
  • Ofelia Banihashemi *, RamazanAli Khavarinejad, Narguess Yassa, Farzaneh Najafi Pages 46-56

    Atropa komarovii generates tropane alkaloids and three other compounds such as hyoscyamine. Racemate atropine and scopolamine (hyoscine) are the main alkaloids that have anticholinergic, antispasmodic and sedative agents. A proficient convention has been reported for the formation of transgenic Atropa komarovii by the use of Agrobacterium rhizogenes. Root culture by utilizing leaves explants that were contaminated by Agrobacterium rhizogenes ATCC 15834, a strain with the paired vector. The hairy roots after contamination for three weeks shaped specifically from the cut edges of leaves. The PCR intensification demonstrated that rol B genes of Ri plasmid of Agrobacterium rhizogenes were coordinated and communicated into the genome of changed hairy roots. Examination of HPLC demonstrated that hairy roots can produce scopolamine and hyocyamine and it was appeared that scopolamine content essentially expanded in changed roots and hyoscyamine extremely expanded in non-transgenic roots. According to the results it was perceived that the scopolamine content in hairy roots was raised significantly since it was compared to control roots. it was evidenced that hairy roots gather a great number of metabolites that have a commercial significance. Thus later on we can enhance efficiency for example by building up the biosynthetic route overexpression of gene codifying enzymes in the metabolic route for expanding valuable secondary metabolites in plant cures.

    Keywords: Atropa komarovii, rol B, Secondary metabolite, Transformation, Tropane alkaloids
  • Atieh Modarresi, Mohsen Nafar, Zahra Sahraei *, Jamshid Salamzadeh, Shadi Ziaie Pages 57-67
    Reduced graft function (RGF) in donor renal transplant recipients is caused by oxidative damage due to extensive ischemia-reperfusion (I/R) injury during transplantation. Neutrophil gelatinase-associated lipocalin (NGAL) is a promising biomarker to detect tubular injury early after renal transplantation. N-acetylcysteine (NAC) is a potent antioxidant that can reduce I/R injury by improving oxidative damage. The aim of the present study is to assess the efficacy of NAC in improving graft function and reducing renal tubular injury in deceased donor renal transplant recipients. A double-blind, randomized clinical trial was conducted on 50 deceased donor renal transplant recipients. Patients were randomized into two groups, receiving either 600 mg NAC twice daily, or placebo (days 0 to 5). Results were assessed based on the rate of RGF, levels of plasma NGAL (p-NGAL) and the estimated glomerular filtration rate (eGFR). The rate of RGF was significantly lower in patients receiving NAC vs. placebo (21.4% vs. 50%). The measurement of p-NGAL levels showed that patients in the NAC group had significantly greater reduction of p-NGAL by both days 1 and 5 post-transplantation than those in the placebo group. A near steady-state eGFR level was reached by week 1 in the NAC group, however, the improvement of eGFR was significantly slower in the placebo group and a near steady-state was only achieved by week 4. NAC has promising potential in reducing tubular injury and improving graft function, evidenced by significant reduction in the rate of RGF and levels of p-NGAL.
    Keywords: Transplants, kidney transplantation, Reperfusion injury, Acetylcysteine, Lipocalin-2, Glomerular Filtration Rate
  • Fatemeh Mohammad Pour, Ava Mnsouri *, Molouk Hadjibabaie Pages 68-76

    Vaccination against HPV seems to be a good approach for prevention of cervical cancer and genital warts. But in Iran we are confronted with lack of evidence for its cost-effectiveness whereas its consumption is dramatically increasing. This was a cross- sectional study. We used a questionnaire included 5 sections as follows; Patients demographic information, Patients medical history, Pregnancy and lactation considerations, Gardasil prescription characteristics and HPV infection characteristics. Prescriber with adherence to guideline were defined as those prescribing Gardasil for correct age range and indication with accurate dosage and administration. Descriptive statistics for variables was shown by frequency (percent) or mean (±SD) and evaluation of relation between categorical variables was performed by using Chi-square test. Total 566 Gardasil recipients participated in the study. There were mostly female with mean age of 28.1 (±6.68). For 128 (22.6%) participants Gardasil prescribed correctly considering both age range (9-26 years) and indication (prophylactic). From this group, 80 participants (14.1%/566) have received accurate Gardasil dosage and interval (prescriber had adherence to guideline). Patients’ out of pocket payment in guidelines adherent prescriptions was a seventh of total costs obtained from 566 consumers. Gynecologists significantly prescribed for prophylaxis higher than other specialties (p-value=0.01). Prescribers’ practice in administrating Gardasil is obviously not appropriate and it is imposing burdensome cost to community and government. On the other hand we encounter with increase rate of use in Iran for past years. Therefor we are in urgent need for appropriate interventions in national level and prompt supervision to regulate Gardasil consumption.

    Keywords: Gardasil, human papilloma virus vaccine, utilization, Iran, guideline adherence
  • Sahar Mofidi Tabatabaei, Mahdi Moridi Farimani *, Samad Nejad Ebrahimi, Peyman Salehi Pages 77-83

    The genus Tanacetum includes some popular endemic species of the flora of Iran, with important medicinal properties. In a project directed at structurally interesting bioactive metabolites from Iranian endemic species, we studied Tanacetum sonbolii Mozaff. Eight compounds comprising six phenolic and two terpenoidal compounds were isolated from the ethyl acetate extract of the aerial parts of the plant by normal and reverse phase chromatography. Their structures were established mainly by 1D and 2D NMR spectroscopic techniques, including 1H-1H COSY, HSQC and HMBC methods and confirmed by comparing their NMR data with those reported in the literature. The compounds namely: 2,4-dihydroxy-6-methoxyacetophenone (1), apigenin (2), 5-desmethylsinensetin (3), 5-desmethylnobiletin (4), 8-methoxycirsilineol (5), scopoletin (6), ursolic acid (7) and β-sitosterol (8). In vitro antiprotozoal activity of compounds 1, 3, and 5 were evaluated against Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum parasites and also toxicity against rat myoblast (L6) cells. Compound 5 showed promising activity against T. b. rhodesiense.

    Keywords: Tanacetum sonbolii, Asteraceae, Flavonoids, Coumarin, antiprotozoal activity
  • Azar Mostoufi, Nargese Chamkouri, Samaneh Kordrostami, Elham Alghasibabaahmadi, Ayyub Mojaddami * Pages 84-97
    AIDS, as a lethal disease, is caused by infection with the HIV virus that affects millions of people. Three essential enzymes should be encoded for replication of HIV virus: protease, integrase and reverse transcriptase (RT). RT has two different activities including DNA polymerase and ribonuclease H (RNase H). However, all of the marketed RT inhibitors target only the DNA polymerase activity. Therefore, ribonuclease H activity may serve as a new target for drug discovery. In the present study, a series of 3-Hydroxypyrimidine-2,4-dione derivatives as potent RT-associated RNase H inhibitors were applied to QSAR analysis. Two methods including multiple linear regressions (MLR) and partial least squared based on genetic algorithm (GA-PLS) were utilized to find the relationship between the structural feathers and inhibitory activities of these compounds. The best multiple linear regression equation was generated by GA-PLS method. A combination of 2D autocorrelations, topological, atom-centered and geometrical descriptors were selected by GA-PLS as they had more effects on the inhibitory activity. Then, the molecular docking studies were carried out. The results showed that the important amino acids inside the active site of the enzyme responsible for essential interactions were Gln475, Asp549, Tyr501, Ser515, Trp534, Asp493, Tyr472 and Gln480 which took part in hydrogen bond formation. Furthermore, docking energy was plotted against pIC50 predicted by GA-PLS method. The result showed that there is a good correlation with R2=0.71. Consequently, these findings suggest that the better method, GA-PLS, could be applied to design new compounds and predict their inhibitory activity.
    Keywords: QSAR, Molecular docking, reverse transcriptase, AIDS, Antiviral Agents
  • Samira Asgharzadeh, Zahra Rabiei, Sana Rabiei, Elham Bijad, Mahmoud Rafieian Kopaei * Pages 98-110

    Prolonged epileptic seizures are the cause of neuronal death and brain damage. Lesions in different regions of the brain can lead to memory loss and cognitive disorders. It is therefore essential to seek out new neuroprotective drugs. Our aim was to investigate the therapeutic effects of oleuropein to improve seizure, oxidative stress and cognitive disorder in pentylenetetrazole (PTZ) kindling mouse model of epilepsy. Mice were randomly assigned to four groups; (1) PTZ group that intraperitoneally received PTZ for 10 days, (2) oleuropein group that received oleuropein (20mg/kg) 30 minutes before PTZ administration, (3) diazepam group that received diazepam 30 minutes before PTZ administration and (4) flumazenil group that was administered with flumazenil and then oleuropein 30 minutes before PTZ administration. Epilepsy severity was determined after final administration of PTZ. Then, the hippocampal tissue was removed and stored at -70°C to measure the interleukin-1 (IL-1) and glutamate transporter 1 (GLT-1) gene expression. Oleuropein treatment caused a significant increase in seizure latency and a significant decrease in total frequencies of head ticks, head and upper limbs seizures, the whole body seizures, frequent spinning and jumping and tonic seizures in PTZ receiving mice. IL-1 expression decreased and GLT-1 levels did not change significantly in oleuropein group. Oleuropein treatment caused significant improvement of passive avoidance memory in PTZ receiving mice in shuttle box test. Oleuropein can decrease PTZ-induced seizures and memory disorders due to its antioxidant and anti-inflammatory properties and thus can be used to produce antiepileptic drugs.

    Keywords: Oleuropein, Seizure, Pentylenetetrazol, Anti-inflammatory, Antioxidant
  • Kaveh Kazemian, Shahram Ala *, Mojtaba Mojtahedzadeh, Mahmoud Abedini, Abbas Alipour, Saeid Abediankenari, Mohammadreza Rafati, Ali Abaskhanidavanloo, Fatemeh Mohajerani Pages 111-119
    Stroke was the second most frequent cause of death after coronary artery disease. The purpose of this study was to evaluate the effects of L-carnitine and Intralipid Fat emulsion alone and in combination in patients with ischemic stroke. In a prospective, RCT, and double-blind study 100 patients with MCA ischemic cerebrovascular accident who were admitted in the first 24 hours of injury entered the study. Patients were randomly assigned into four groups of L-carnitine, Fat emulsion, L-carnitine plus Fat emulsion and control. 500 ml of Fat emulsion 10% was infused over 6 to 12 hours and 1 gr of l-carnitine was given orally to patients in addition to common therapies, according to the (AHA/ASA) guidelines. Blood samples before the intervention, then after 24 hours, 48 hours and 7 days later were taken and immunoenzymatic colorimetric method was used for quantitative determination of S100B concentration in patient’s serum. All of our treatment interventions have decreased S100B levels statistically significant (p < 0.05), Changes in observed levels of S100B before and after intervention were different between the groups and the observed differences were statistically significant (p= 0.01). In the GEE model, it was found that S100B levels in the L-carnitine plus Fat emulsion group decreased more than the control group and this decline has been statistically significant [20.47 (CI 95%: 6.25-34.41)]. Based on the results obtained from this study, it seems that L-carnitine with Intralipid Fat emulsion could lead to neuroprotective effects with a significant reduction in the S100B biomarker
    Keywords: l-carnitine, fat emulsion, Neuroprotection, CVA, Biomarker
  • Rezvan Dehdari Vais, Hossein Yadegari, Hossein Heli * Pages 120-137
    An electrochemical sensor was designed and fabricated for electrocatalytic oxidation and determination of famotidine in pharmaceutical forms. The electrochemical oxidation process and its kinetics were investigated using cyclic voltammetry, steady-state polarization measurements, and chronoamperometry techniques, and also the analytical measurements were performed by amperometry. Upon addition of the drug into the solution, cyclic voltammograms of the fabricated sensor exhibited an increased anodic peak current associated with a decrease in the corresponding cathodic current. These results suggested an electrocatalytic EC’ oxidation mechanism for famotidine on the oxyhydroxide species immobilized on the electrode surface. Accordingly, a mechanism involving generation of Ni3+ active sites and their subsequent consumption by the drug was proposed. Moreover, the corresponding rate law under the control of charge transfer was developed and kinetic parameters were derived. A sensitive and time-saving amperometric procedure was also developed for the analysis of famotidine with a detection limit of 5.91 mmol L-1. Using the developed amperometric procedure, famotidine was successfully analyzed in the presence of ibuprofen. The developed sensor in this study displayed enhanced sensitivity and selectivity, compared to some other reported methods.
    Keywords: Nickel, Pepcid, Nanoflower, Nanomaterial, Electrocatalysis, Electroanalysis
  • Ericka Anguiano Almazan, Pablo Serrano Castañeda, Roberto Diaz Torres, Jose Escobar Chavez * Pages 138-152

    The development of a losartan potassium patch for the treatment of hypertension established that a combination of hydrophobic and hydrophilic polymers, using as a plasticizer citroflex and succinic acid as a cohesion promoter result in homogeneous films. The effect of the Eudragit® E100, PVP K30, citroflex and succinic acid in the bioadhesion, postwetting bioadhesion, resistance to rupture and drug release, was studied. The succinic acid in synergy with the plasticizer (citroflex) modifies the characteristics of the polymeric matrix of Eudragit® E100, increasing the release and the resistance to rupture of transdermal patches. For the case of the hydrophilic polymer PVP K30, it increases the bioadhesion and drug release by creating porous matrices. From a previous experimental design, the optimal formulation was acquired, this formulation was characterized physicochemically. A transdermal patch was obtained with the next dimensions and characteristics: 28.46 ± 0.055 mm in diameter and 0.430 ± 0.008 mm in thickness, a bioadhesion of 1063.05 ± 60.33 gf, postwetting bioadhesion 995.9 ± 72.53 gf significantly decreased. The breaking strength was of 1301.5 ± 96.5 gf, surface pH patch of 6, constriction of 0% at 7 days, and 94.0366 ± 1.8617% of losartan content. The 93% of the drug is released at 4 h (n = 6), adjusting to the kinetic model of Higuchi and Peppas. In the in-vitro penetration studies by passive diffusion, a flow (J) of 42.2 μg/cm2h, a permeability constant (kp) of 2.1793E-03 cm/h and a latency time (tL) of 17.20 h and with the use of microneedles a flow (J) of 61.7 μg/cm2h, a permeability constant (kp) of 3.1869E-03 cm/h and a latency time (tL) of 17.74 h were obtained.

    Keywords: Transdermal patch, Solid microneedles, Polyvinylpyrrolidone, skin, Losartan, Eudragit E100®
  • Mohammad Sayyadi, Ava Safaroghli Azar, Majid Safa, Hassan Abolghasemi, Majid Momeny, Davood Bashash * Pages 153-165

    Due to the frequent contribution in the pathogenesis of different human malignancies, c-Myc is among those transcription factors that are believed to be pharmacologically targeted for cancer therapeutic approaches. In the present study, we examined the anti-leukemic effect of a well-known c-Myc inhibitor 10058-F4 on a panel of hematologic malignant cells harboring either mutant or wild-type p53. Notably, we found that the suppression of c-Myc was coupled with the reduction in the survival of all the tested leukemic cells; however, as far as we are aware, this study suggests for the first time that the cytotoxic effect of 10058-F4 was not significantly affected by the molecular status of p53. Delving into the molecular mechanisms of the inhibitor in the most sensitive cell line revealed that 10058-F4 could induce apoptotic cell death in mutant p53-expressing NB4 cells through the suppression of NF-κB pathway coupled with a significant induction of intracellular reactive oxygen species (ROS). In addition, we found that the anti-leukemic effect of 10058-F4 was overshadowed, at least partially, through the compensatory activation of the PI3K signaling pathway; highlighting a plausible attenuating role of this axis on 10058-F4 cytotoxicity. In conclusion, the results of the present study shed light on the favorable anti-leukemic effect of 10058-F4, especially in combination with PI3K inhibitors in acute promyelocytic leukemia; however, further investigations should be accomplished to determine the efficacy of the inhibitor, either as a single agent or in a combined-modal strategy, in leukemia treatment.

    Keywords: 10058-F4, c-Myc, p53, NF-κB pathway, PI3K pathway, Autophagy
  • Mahmood Barani, Masoud Torkzadeh Mahani *, Mohammad Mirzaei, MohammadHadi Nematollahi Pages 166-180

    An efficient gene delivery system has some critical factors that enhance the efficiency of nanocarrier. These factors are low production cost, high bioavailability, high encapsulation efficiency, controllable release, and targeting ability. Niosome (the nonionic surfactant vesicles) was considered as a promising gene delivery system. Niosome can increase stability and uptake of active agents. We used all mentioned factors in one optimized formulation entitled plasmid- loaded magnetic niosomes (PMN). To increase the bioavailability of niosomes, we used ergosterol (a natural lipid) instead of cholesterol in structure of niosome. Also, cetyl trimethyl ammonium bromide (CTAB) in different concentrations was used to improve encapsulation of plasmid and compared to niosomes that did not have CTAB (negative niosome). Afterward, magnetic nanoparticle (Fe3O4@SiO2) was synthesized and loaded into niosome to obtain targeting ability. Prepared formulations were evaluated regarding size, zeta potential, morphology, encapsulation of magnetic nanoparticles and plasmid (Pm-cherry-N1), release rate, and transfection efficiency.  Results demonstrated that optimum formulation (Nio/CTAB3%/Fe/P) has a nanometric size (118 ± 2.31 nm, positive zeta potential (+25 ± 0.67 mV), high loading of plasmid (72%), and good gene expression (35%). Interestingly, after applying a magnetic field below the cell plate, we obtained ac increased gene expression from 35% to 42%. These results showed that this new formulation would have a promising future and also can be used for delivering the other drugs and active agents.

    Keywords: Gene expression, Targeting, Niosome, Magnetic nano particle, Plasmid
  • Zahra Haghighijoo, Omidreza Firuzi, Savis Meili, Najmeh Edraki, Mehdi Khoshneviszadeh *, Ramin Miri Pages 181-191
    Among Recent advances in the identification of anti-inflammation agents, anti-cytokines (like Interleukin-1), related to p38 MAPK families play an important role; Here in we designed new effective and low toxic anti-cytokine agents based on 1-Hydroxy-2,4,5-triaryl imidazole derivatives. The reaction of oximoinoketone intermediate with ten different aromatic aldehyde and ammonium acetate in refluxing acetic acid condition give imidazole derived product, the IL-1β inhibitory assay were performed on Human PBMCs (peripheral blood mononuclear cells) using an enzyme-linked immunosorbent assay (ELISA) kit and then in computational part the binding mode of the best compound was accomplished  by docking in Crystal structure of p38 MAP kinase (PDB ID: 1A9U) compared with SB202190 as standard drug. All compounds were synthesized and evaluated in biological assay showing the inhibitory activity from 28% to 82% compared to SB202190 and binding mode analysis revealed that the hydrogen-bond interactions with residues (Met109, Val30) were key point in inhibitor binding. Compound 5g clearly proved the best inhibitory action and could be further utilized for designing newer anti-cytokine agents and p38α MAP kinase potentially inhibitory action.
    Keywords: Anti-cytokine, IL-1β, Imidazole inhibitor, inflammation, Interleukin, P38 MAPK
  • Saeed Dezianian, Jamil Zargan *, HamidReza Goudarzi, Ashkan Haji Noormohamadi, Mohsen Mousavi, Hani Keshavarz Alikhani, Behrooz Johari Pages 192-202

    Scorpion venoms contain potentially useful pharmacological agents. Several studies demonstrate that the venoms of some scorpions induce apoptosis and inhibit the growth of cancer cells; therefore, they have been investigated for isolating anticancer components. In this study, antitumor effects of Hottentotta schach crude venom on MCF-7 (breast cancer cell line) as test group and Vero (African green monkey kidney normal cell line) as control group were analyzed. Cell toxicity was analyzed using MTT and neutral red (NR) uptake assays and apoptosis induction was analyzed using comet assay and caspase-3 activity. Oxidative stress following Hottentotta schach crude venom treatment was analyzed using nitrite oxide (NO) determination assay, reduced glutathione (GSH) and catalase enzyme activity assays. Results showed that crude venom (25-200 μg/mL) induced apoptosis and inhibited the growth of MCF-7 and to a lesser extent in Vero cell lines. Nitrite oxide concentration increased while glutathione concentration and catalase enzyme activity were decreased in MCF-7 cells; however, results in Vero cells were reversed completely. It can be concluded that Hottentotta schach crude venom disturbs the oxidation and reduction potential in cancer cells and ultimately induce apoptosis. So this venom can be used as a good source for isolation and designing new anticancer drugs.

    Keywords: Cancer therapy, Anti-cancer drug, Hottentotta schach, Scorpion crude venom
  • Gita Afsharmanesh, Gholamhossein Mehralian *, Farzad Peiravian Pages 203-217

    Discrete choice experiments (DCES) as a stated preference method have used increasingly todetermine preferences attached to some attributes associated to health. Although, the validity ofthis type of studies comprehensively depends on the appropriate determination of attributes andattribute-levels for DCES, there is little rigorous evidence regarding which factors or attributes andattribute- levels should be counted for eliciting public preferences in health resource allocation.This paper responds to such question by carefully doing a qualitative study. A qualitative studyused semi-structured interviews, which were audio recorded, transcribed and subject to thematicanalysis. Sixteen participants had been key informants and decision makers of pharmaceutical andhealth system. Initially, by conducting a meticulous literature review, an inclusive list of attributesassociated with intended policy was identified. Qualitative data for the development of attributesand their levels were collected using 16 key informant interviews and were analyzed by softwareMAXQDA followed by a focus group discussion (FGD) with 7 people, well-familiar with thenotion pharmaceutical policy and Pharmacoeconomics. The 311 codes in four main dimensionswere initially identified by conducting interviews. However, for being manageable within aDCE, they were classified and limited to four attributes, including severity of disease withouttreatment, health gain after treatment, frequency of patients, and cost of treatment per patient.This qualitative study provides enough evidence for designing and doing a precise discrete choiceexperiment answering the question about public preferences in pharmaceutical subsidization andcontributes empirical evidence to the limited methodological literature on attributes developmentfor DCE, specifically within low and middle-income countries.

    Keywords: Discrete choice experiment, Attribute, attribute-level, qualitative research, Pharmaceutical subsidization
  • Tiegsti Bahta, Aman Karim *, Gomathi Periasamy, Gereziher Gebremedhin, Najeeb Ur Rehman, Helen Bitew, Kalay Hagazi Pages 218-230

    Otostegia fruticosa is traditionally used to treat tonsillitis, stomach ache, asthma, arthritis, and febrile illness in different parts of Ethiopia and other countries. In this experiment 70% ethanolic crude extract and fractions of the leaf of Otostegia fruticosa (Forssk.) Schweinf. ex Penzig were evaluated for their in-vivo anti-inflammatory and analgesic activities and in-vitro hyaluronidase inhibition properties at different concentrations. Tail immersion, acetic acid induced writhing and carrageenan-induced paw edema model were used to assess the in-vivo analgesic and anti-inflammatory activities, respectively. Swiss albino mice of either sex were randomly divided into five groups of six mice per group and for evaluation of the fractions randomly divided into six groups of six mice per group. The test groups were treated with hydroalcoholic extract of O. fruticosa at doses of 100, 200, and 400 mg/kg. The positive control groups received either pethidine 5 mg/kg or aspirin at 100 mg/kg or 150 mg/kg. The negative control groups were orally given sunflower oil. All the fractions were administered at the dose of 400 mg/kg. In all models, the higher dose (400 mg/kg) of the crude extract and chloroform fraction showed a significant central and peripheral analgesic and anti-inflammatory activities with comparable effects to standards used.  The hyaluronidase inhibition assay result showed that the test samples displayed concentration-dependent inhibitory activities. These findings indicate that 70% ethanol extract and organic solvent fractions of O. fruticosa leaves have potential analgesic, anti-inflammatory, and enzyme inhibitory activities.

    Keywords: Analgesic activity, anti-inflammatory activity, Otostegia fruticosa, Tail immersion test, Acetic acid- induced writhing, Hyaluronidase inhibition
  • Nahid Ghaedi, Iran Pouraboli *, Nayereh Askari Pages 231-250
    Levisticum officinale (Apiaceae) is a favorite food spice. Iranian folk medicine claims that it has a prominent antidyslipidemic property but this is not documented scientifically so far. This study evaluated this and the other antidiabetic aspects of the stem and leaf hydroalcoholic extract of it (LOE). Regarding to oral glucose tolerance test results, LOE (500 mg/kg) administration 30 min before glucose loading significantly decreased the blood glucose level (13%) at 90 min in male rats. Additionally LOE treatment (500 mg/kg, orally, once a day) for 14 days significantly reduced the serum glucose level (24.97%) and markedly improved the lipid profile and the insulin, creatinine, alanine aminotransferase and aspartate aminotransferase serum levels in diabetic rats. Moreover, LOE effectively amended the impaired antioxidant status and ameliorated lipid peroxidation in the plasma and pancreas and liver tissues of diabetics. Also, 14 days LOE treatment, significantly decreased the renal sodium-glucose cotransporter 2 and facilitated glucose transporter 2 (GLUT2) mRNA levels and GLUT2 gene expression in the enterocytes of jejunum tissue in comparison with diabetic untreated rats. HPLC method revealed the presence of chlorogenic acid, rosmarinic acid, caffeic acid, quercetin and luteolin and GC-MS analysis detected bioactive compounds like phthalides, thymol, phytol, hexanoic acid, carene and menthofuran. LOE showed α-amylase (αΑ) inhibitory activity and in silico studies predicted that among extract ingredients luteolin, quercetin, rosmarinic, caffeic and hexanoic acids have the greatest αΑ inhibition potecy. Thus, current results justify antidyslipidemic value of L. officinale and shed light on more antidiabetic health benefits of it.
    Keywords: Diabetes mellitus, Levisticum officinale, antioxidants, Glucose transporters, Phytochemicals, Molecular docking
  • Reza Araqi, Bi Bi Fatemeh Mirjalili *, Leila Zamani, Soghra Khabnadideh, Kamiar Zomorodian, Zeinab Faghih, Hamhdreza Arabi Pages 251-259
    In order to expand the application of Fe3O4@SiO2-SnCl4 in the synthesis of heterocyclic compounds, in this study, we wish to report the use of one-pot three component synthesis of pyrimido[4,5-b]quinolone derivatives (D1-D16) through reaction of 6-amino-2-(methylthio)pyrimidin-4(3H)-one, dimedone, or 1,3- cyclohexadione and aldehydes in the presence of Fe3O4@SiO2-SnCl4 as an efficient eco-friendly catalyst under ultrasound irradiation. The final aim of this study is evaluation of antifungal activity of resulted products.
    Synthesis of pyrimido[4,5-b]quinolin derivatives were done via three components coupling reaction of aldehyde, dimedone or 1,3-cyclohexadione and 6-amino-2-(methylthio)pyrimidin-4(3H)-one in the presence of Fe3O4@SiO2-SnCl4 under ultrasonic irradiation in water at 60 oC. The products structure were studied by FT-IRI, 1H NMRII and 13C NMRII. All the compounds were screened for antimicrobial activity by broth microdilution methods as recommended by CLSIIII.
    Considering our results showed that compound (D13) had the most antifungal activity against C. dubliniensis, C. Albicans, C. Tropicalis and C. Neoformance at concentrations ranging (MIC90) from 1-4 μg/mL. Compounds (D9), (D10), (D14) and (D15) had significant inhibitory activities against C. dubliniensis at concentrations ranging (MIC90) from 4-8 μg/mL respectively.
    5-(3,4-dihydroxyphenyl)-8,8-dimethyl-2-(methylthio)-5,8,9,10 tetrahydropyrimido[4,5-b]quinoline-4,6(3H,7H)-dione (D13) exhibited inhibitory and fungicidal activities against the tested yeasts. The specific binding mode or the binding orientation of more efficient compounds to CYP51 active site, have been also performed by molecular modeling investigations and showed that there is a good correlation with biological test.
    Keywords: Pyrimido[4, 5-b]quinolin, Antifungal activity, heterogeneous solid acid, Fe3O4@SiO2-SnCl4, Molecular modeling
  • Afsoon Akbarzadeh, Pezhman Sasanpour, HamidReza Moghimi * Pages 260-270

    LED light is used for many medical and cosmetic applications such as phototherapy and skin rejuvenation. Such physical methods can be combined with drug therapy, such as LED-responsive drug delivery system, the subject of present investigation. To perform this investigation, a nanoliposome composed of DPPC, DSPE-PEG2000 and DC8,9PC, was prepared as LED-sensitive systems. Calcein was loaded in the liposomes as a fluorescent probe for drug release studies. Different LED wavelengths (blue, green and red) were used for triggering release of calcein from nanoliposome. Indoor daylight, darkness and sunlight were applied as controls. Results showed that liposomes do not release their cargo in darkness, but they released it in response to indoor daylight, sunlight and LEDs, with the blue light showing the highest effect. Results also showed that release of calcein was sensitive to wavelength. Our results reveal potential of LED-sensitive liposomes for medical and cosmetic applications and that such system can be combined with phototherapy. Such concomitant therapies can increase medical/cosmetic effects and decrease adverse reactions to phototherapy.

    Keywords: Liposome, LED light, Trigger release, Controlled release, Drug delivery
  • Fateme Azemati, Bahman Jalali Kondori *, Hadi Esmaeili Gouvarchin Ghaleh Pages 271-281
    Although hydroxyurea is one of the most widely used drugs in treating breast cancer, the use of it leads to some side effects. Hence, in order to reduce complications of treatment and increase its efficiency, drug delivery has been attracted more attention. Present study included three stages. The first stage was involved in the synthesis of nanoparticles-loaded hydroxyurea that its characteristics were evaluated by using scanning electron microscopy and Zetasizer system. In the second stage, cultured MCF-7 cells were undergone treatments by hydroxyurea and Nanoparticles-loaded  hydroxyurea in various concentrations. In the third stage, the MCF-7 was treated by IC50 of hydroxyurea and nanoparticles-loaded hydroxyurea which are in combination with radiation and hyperthermia. Afterward, the viable of cell, apoptosis, and levels of caspase-8 and-9 proteins were assessed. The average size and the potential surface of nanoparticles and nanoparticles-loaded hydroxyurea were 26 nm, 48 nm, 3.86 mV, and -29.3 mV, respectively. Results of MTT assay and apoptosis represented that the percentage of cytotoxicity in the treated groups by in combination group and nanoparticles-loaded hydroxyurea was significantly increased in comparison with hydroxyurea. This increase was dependent on the concentration of nanoparticles-loaded hydroxyurea. Nevertheless, the activity of caspase-8 shows any significant changes, the activity of caspase-9 was significantly increased in the control and treatment groups. We concluded that nanoparticles-loaded hydroxyurea and it in combination with radiation and hyperthermia have higher toxicity effect on MCF-7 cells in comparison with pure hydroxyurea and induced mitochondrial-dependent apoptosis by down-regulation of caspase-8 and up-regulation of caspase-9 expressions.
    Keywords: Hydroxyurea, Fe3O4 NP, polyethylene glycol, MCF-7 Cells, Cytotoxicity, Radiation, Hyperthermia
  • Nabila Zaabat, Anne Emmanuelle Hay, Serge Michalet, Inès Skandrani, Leila Chekir Ghedirac, Marie Geneviève Dijoux Francaa, Salah Akkal * Pages 282-291

    In the present work, chemical investigation of the aerial parts of Phlomis bovei de Noé an endemic species from Algeria, led to the isolation and identification of seven known compounds including five flavones glycosides: Chrysoeriol 7-O-(3’’-(E et Z)-p-coumaroyl)-β-glucoside (1), terniflorin (apigenin-7-O-(6’’-E-p-coumaroyl)glucoside) (3), apigenin-7-O-(6’’-(5’’’-methoxy-coumaryl) glucoside (4), apigenin 7-O-(3″-p-coumaryl)glucoside(5), hispidulin-7-O-glucuronide (6) and two cinnamic acid derivatives: p-coumaric acid methyl ester (E et Z) (2), chlorogenic acid (7). Compound 4 is described for the first time in the species bovei de Noé , the genus Phlomis and the Lamiaceae family. Structures elucidation was performed by comprehensive 1D and 2D NMR analyses, mass spectrometry and by comparison with literature data. Some pure compounds and extracts have been evaluated for their antioxidant activities through different


    DPPH and ABTS assays as well as CUPRAC assay. Genotoxic and antigenotoxic activities of pure compounds were also evaluated in-vitro on Escherichia coli PQ37 cells by the SOS Chromotest.

    Keywords: Phlomis bovei De Noe, Chemical constituents, Antioxidant, Genotoxic effect, Antigenotoxic activity
  • Ameeduzzafar Zafar, Nazia Khan, Nabil K Alruwaili, Syed Nasir Abbas Bukhari, Bader Alsuwayt, Muhammad Afzal, Sana Akhtar, Mohd Yasir, Mohammed Elmowafy, Khaled Shalaby, Asgar Ali * Pages 292-311
    Conjunctivitis is considered as a common infection of ocular surfaces. Eye drop is most commonly used for treatment of conjunctivitis, but has some drawback like 95% drug eliminated after administration. Administration of levofloxacin to the anterior site in form of chitosan coated poly (lactic-co-glycolic acid) nanoparticles (LFV-CS-PLGA-NPs) expected to overcome these problem and increasing corneal contact time and permeability for effective treatment of bacterial conjunctivitis. TheNanoparticles were developed by single emulsion solvent evaporation technique and optimized for different variables (chitosan, poly (lactide-co-glycolic acid) and polyvinyl alcohol concentration) by employing three factors, three levels Box–Behnken statistical design. The nanoparticles were evaluated for particle size, drug loading, entrapment efficiency, drug release, ex-vivo permeation, ocular tolerance, antimicrobial study, confocal laser microscopy, and Gamma scintigraphy study. The particle size and PdI of the optimized nanoparticles were 169.968 ± 15.23 nm and 0.13 ± 0.03, respectively, where as entrapment efficiency and drug loading is 49.54 ± 2.43% and 11.29 ± 2.13% with extended release profile and strong mucoadhesion. DSC data indicated levofloxacin formed molecular dispersion within coated nanoparticles. Corneal flux showed significantly (P < 0.05) higher permeation as compared to marketed formulation. Formulation was nonirritant and possessed good antibacterial activity. Gamma Scintigraphy showed slow drainage compared to drug solution, indicating reduction in nasolachrymal drainage. The Gamma Scintigraphy study indicated the CS coated PLGA-NPs have high corneal residence time as compared to drug solution. So, it is revealed that LFV-CS-PLGA-NPs increase the drug concentration over ocular tissue and potential usefulness for sustained drug delivery.
    Keywords: Levofloxacin loaded coated PLGA nanoparticles, Box-Behnken, Ocular irritation, Confocal microscopy antimicrobial study, Gamma scintigraphy
  • Nasser Gholijani *, Samira Sadat Abolmaali, Kurosh Kalantar, MohammadHadi Ravanrooy Pages 312-320

    Rheumatoid arthritis (RA) is one of the most common autoimmune diseases. Carvacrol,an important natural terpenoid product in aromatic plants such as thyme, has shown antiinflammatoryeffects in animal models of arthritis. However, its poor water solubility andhigh volatility have limited its application. In the present study in order to overcome thisproblem, we encapsulated carvacrol in the bovine serum albumin (BSA) nanoparticles andexamined its therapeutic and immunomodulatory effects in adjuvant-induced arthritis (AIA).Carvacrol-loaded BSA nanoparticles were prepared by desolvation method. Nanoparticleshad encapsulation efficiency (EE) of 67.7 ± 6.9% and loading capacity (LC) of 26.6 ± 2%.The size of particles was 148 ± 25 nm and they had monomodal distribution. After arthritisinduction, the rats were treated intraperitoneally with nanoparticle for every 3 days until day28. The treatment of the rats with 375 mg/mL carvacrol-loaded BSA nanoparticle significantlydecreased clinical severity score (27.5 ± 9.8%, p = 0.008), erythrocyte sedimentation rate(33.4 ± 10%, p = 0.02), nitric oxide production (82.3 ± 2.6%, p = 0.004) and interleukin (IL)-17 gene expression (55.1 ± 8.2%, p = 0.003) compared to the untreated arthritic group. Ahigher reduction in inflammation severity in arthritic rats treated with carvacrol-loaded BSA incomparison to those treated with carvacrol alone was observed. In conclusion, encapsulationof carvacrol in nanoparticles reduced arthritis signs and release of NO and IL-17 inflammatorycytokine and therefore is suggested to be considered as a good approach for improving thetherapeutic applications of carvacrol in RA.

    Keywords: Carvacrol, Albumin nanoparticles, Adjuvant-induced arthritis, Anti-inflammatory effect, Rheumatoid Arthritis
  • Josiane Elia, Karina Petit *, Jean Michel Huvelin, Mona Tannoury, Mona Diab Assaf, Delphine Carbonnelle, Hassan Nazih Pages 321-330

    Marine macroalgae have attracted much attention in recent years as a valuable source of bioactive metabolites. The cytotoxic potential of the Laurencia papillosa red alga collected from the Lebanese coast has been investigated on human breast cancer cells MCF-7. The crude extract of Laurencia papillosa (L. papillosa) was fractionated by column chromatography using a series of increasingly polar solvents (methylene chloride, acetone and methanol). Cytotoxicity of the crude extract and fractions was determined by MTT assay in MCF-7 cells. Apoptosis was detected by annexin V/propidium iodide assay and by measurement of Bcl-2 expression. Flotillin-2 expression was examined using RT-qPCR and Western blot. The crude extract, and the fractions of CH2Cl2 and acetone exhibited a dose-dependent cytotoxic effect on MCF-7 cells. Apoptosis was specifically induced by one of the acetone fractions having the highest cytotoxicity. It has been demonstrated by an increase in late phase apoptotic cell populations, and a decrease in Bcl-2 anti-apoptotic marker expression on mRNA and protein levels in a dose- and time- dependent manner. Furthermore, this active fraction decreased Flotillin-2 expression associated with cancer progression. Our data suggest that L. papillosa is an important source of cytotoxic metabolites. Further studies are needed for the chemical characterization of the metabolite associated with observed biological activities.

    Keywords: Laurencia papillosa, MCF-7 Cells, Cytotoxic activity, Apoptosis, Flotillin-2
  • Gelareh Vahabzadeh *, Nahid Rahbar Roshandel, Soltan Ahmad Ebrahimi Pages 331-342

    In the present study, we investigated the effects of noscapine (0.5-2 µM), an alkaloid from the opium poppy(Papaver somniferum), on primary murine cortical neurons exposed to 60 min oxygen–glucose deprivation (OGD) in the presence of 5 µM BD-1047, a selective sigma-1 receptor antagonist. The experiments were performed on cortical neurons after 11–16 days of culture. To initiate oxygen–glucose deprivation, the culture medium was transferred to glucose-free DMEM, and placed in a humidified incubation chamber containing a mixture of 95% N2 and 5% CO2 at 37 °C for 60 min. In order to explore the effect on neurons under oxygen–glucose deprivation in this condition, some cultures were pretreated with noscapine and BD1047 together, 24 h prior to OGD followed by 24 h recovery. Cell viability, nitric oxide (NO) production and intracellular calcium concentration ([Ca2+]i) levels were evaluated by MTT assay, the modified Griess method, and Fura-2, respectively. Pretreatment of the cultures with noscapine in the presence of BD1047 significantly increased cell viability and decreased NO generation in a dose-dependent manner compared to BD1047 alone. Pretreatment with 2 μM noscapine and BD-1047 was shown to decrease the rise in [Ca2+]i induced by sodium azide (NaN3) and glucose deprivation. We concluded that noscapine in the presence of BD1047 could protect primary cortical neurons after oxygen–glucose deprivation-induced cell injury but this effect was not complete. Our results indicate that neuroprotective effects of noscapine could be mediated partially through activation of sigma-1 receptor and by decreasing NO production and [Ca2+]i levels.

    Keywords: Noscapine, NO, Oxygen-Glucose Deprivation, Cortical culture, Sigma receptor, Intracellular calcium
  • Ravieh Golchoobian, Fatemeh Nabavizadeh *, Mehrdad Roghani, Alireza Foroumadi, Maryam Izad, Maryam Bahrami, Hafseh Fanaei Pages 343-354
    MDMA (3,4-methylenedioxymethamphetamine, ecstasy) is often abused by youth as a recreational drug. MDMA abuse is a growing problem in different parts of the world. An important adverse consequence of the drug consumption is hepatotoxicity of different intensities. However, the underlying mechanism of this toxicity has not been completely understood. Ghrelin is a gut hormone with growth hormone stimulatory effect. It expresses in liver, albeit at a much lower level than in stomach, and exerts a hepatoprotective effect. In this study, we investigated hepatotoxicity effect of MDMA alone and its combination with ghrelin as a hepatoprotective agent. MDMA and MDMA+ ghrelin could transiently increase serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) followed by tissue necrosis. However, they could significantly decrease liver tumor necrosis factor-a (TNF-α) in both treatment groups. Unexpectedly, in MDMA treated rats, Bax, Bcl-xl, Bcl-2, Fas, Fas ligand (Fas-L), caspase 8, cytochrome c, caspase 3 gene expression, and DNA fragmentation were nearly unchanged. In addition, apoptosis in MDMA+ ghrelin group was significantly reduced when compared with MDMA treated animals. In all,MDMA could transiently increase serum transaminases and induce tissue necrosis and liver toxicity. Ghrelin, however, could not stop liver enzyme rise and MDMA hepatotoxicity. MDMA hepatotoxicity seems to be mediated via tissue necrosis than apoptotic and inflammatory pathways. Conceivably, ghrelin as an anti-inflammatory and anti-apoptotic agent may not protect hepatocytes against MDMA liver toxicity.
    Keywords: hepatotoxicity, 3, 4-methylenedioxymethamphetamine, Ghrelin, TNF-α, Apoptosis, Necrosis
  • Vahideh Montazeri, MohammadHossein Ghahremani, Hamed Montazeri, Mandana Hasanzad, Majid Safavi, Mohsan Ayati, Mohammad Chehrazi, Baharak Arefi Moghaddam, Seyed Nasser Ostad * Pages 355-365

    One of the main genotoxic drugs used in bladder cancer chemotherapy is cisplatin. While it is applied in most types of cancers, resistance to cisplatin is wildly common. In order to overcome drug resistance, it is necessary to determine a predictive marker. This study was conducted to provide basic data for selecting and designing a gene profile for further cohort and RCT studies in the future to improve response to treatment in bladder cancer. The expression levels of ERCC1, MLH1, MSH2, and CTR1 mRNA were determined in the tumor tissue using real-time q-PCR. Progression-free survival (PFS) was analyzed in term of the level of genes expression. The results revealed that the level of ERCC1 mRNA expression was higher in the recurrence (R) group compared to the no recurrence (NR) group. Moreover, the PFS time was increased in the patients with an ERCC1 expression level of below 1.57. The level of MLH1 and MSH2 mRNA expression was lower in the R group compared to the NR group; therefore, PFS time was increased in the patients with MLH1 and MSH2 gene expression levels above the cutoff point. While the level of CTR1 mRNA expression was higher in the R group versus the NR group, the PFS time was longer in the patients with CTR1 expression levels of below 1.265 compared to the patients with high levels of CTR1 expression. It can be concluded that the level of ERCC1, MLH1, MSH2, and CTR1 mRNA expression may be associated with PFS time as possible therapeutic targets for decreasing cisplatin resistance.

    Keywords: carcinoma, Transitional cell, Progression-free survival, Copper transporter 1 protein, Human, Cisplatin