Tehran University Medical Journal

Background: Numerous studies have shown the protective effects of saffron against oxidative damage in a global model of cerebral ischemia, but its effects on brain edema and oxidative ischemic injury in focal ischemic stroke are not completely understood. Therefore, this study was designed to investigate the effects of saffron on brain edema, infarct volume, antioxidant enzyme activity (glutathione peroxidase and superoxide dismutase) and concentration of malondialdehyde (MDA) in ischemic brain tissue in an experimental model of stroke.

Methods: Focal brain ischemia was established with the temporary occlusion of the middle cerebral artery for one hour in rats. Saffron (100 mg/kg) was given intra-peritoneally at the onset of ischemia. 24 hours later, brain edema and infarct volume were evaluated and glutathione peroxidase and superoxide dismutase activities and MDA concentration were measured in the ischemic brain tissue using a specific kit.

Results: The results showed that saffron reduced infarct volume by 77% (P<0.001) and brain edema by 60% (P<0.001) compared with the control group in 24 hours following ischemia. Moreover, saffron significantly reduced the content of MDA (P<0.001) and increased the activity of superoxide dismutase (P<0.001) and glutathione peroxidase (P<0.001) in the cortex of the ischemic brain tissue.

Conclusion: Saffron has protective effects against oxidative ischemic damage and brain edema in a transient model of focal cerebral ischemia in rats. This protective effect is probably induced by increasing the capacity of antioxidant enzymes and decreasing the production of free radicals.

Background: Anterior cruciate ligament (ACL) reconstruction is a first choice treatment for ACL-deficient knees, and arthroscopic single-bundle reconstruction has been widely accepted around the world in this regard. Although, such single-bundle reconstructions result in sufficient knee stability in most cases, but some patients have not been satisfied with postsurgical results in both short-term and long term clinical studies. One of the reasons for these unsatisfactory results could be related to the fact that normal function of the native ACL has not been restored by the traditional ACL reconstruction which uses only a single-bundle graft. The natural ACL consists of a 3-dimensional structure with multibundle fascicles, which can be anatomically divided into 2 main bundles, the anteromedial (AM) and the posterolateral (PL) bundles named for the orientation of their tibial insertions. The purpose of the present study was to compare double-bundle and single-bundle ACL reconstruction.
Methods: Twenty-two patients with anterior cruciate ligament tear who were candidates for ACL reconstruction were enrolled in the study undertaken in Shariati Hospital from 2009 to 2010. Fourteen patients underwent single-bundle and 8 patients double-bundle ACL reconstruction. The patients were evaluated by arthrometer and physical examination in postoperation follow up visit at least 9 months after the operations.
Results: Four out of 14 patients with single-bundle reconstruction had knee joint translation greater than 5 mm but nobody had knee translation more than 5 mm in the double-bundle reconstruction group.
Conclusion: Double-bundle ACL reconstruction seems to be more stable than single-bundle ACL reconstruction.

Background: Bone Marrow Transplantations (BMT) are limited by low CD34+ cell counts in umbilical cord blood (UCB) and these cells need to be expanded for success in such procedures. To achieve this goal, ex vivo expansion of hematopoietic stem cells (HSCs) by enhancing their self-renewal activity on demineralized bone matrix (DBM) scaffold coated with mesenchymal progenitor cells (MPCs) and unrestricted somatic stem cells (USSCs) was recommended. TGF-b pathway is a key inhibitory factor for HSCs self-renewal. In this study ex vivo expansion and downregulation of TGF-b pathway were simultaneously performed.

Methods: USSC cells were isolated from UCB and then coated on DBM scaffold as a feeder layer. UCB CD34+ cells were isolated from UCB by magnetic activated cell sorting (MACS) method and were transfected by siRNA against TGFbR2 in two-dimensional (2D) and three-dimensional (3D) cultures by co-cultivation with USSC. TGFbR2 expression levels were evaluated by quantitative real-time PCR. Cell count and flow cytometry were performed and clonogenic activity was evaluated.

Results: Ex vivo expansion of CD34+ cells was significantly enhanced (41±0.7 folds) by TGFbR2 downregulation, especially in 2D than 3D cultures. Finally, 2D culture showed less TGFbR2 expression levels and higher increase in the percentage of CD34 markers by flow cytometry assay.

Conclusion: The 3D siRNA delivery system would be of lower efficiency in contrast to 2D settings where the cells have less freedom and are in more contact with the feeder layer.

Background: Perioperative administration of tranexamic acid (TA), decreases bleeding and the need for transfusion after cardiac procedures. Hence, the results may vary in different clinical settings and the most appropriate timing to get the best results is unclear. The primary objectives of the present study were to determine the efficacy of TA in decreasing chest tube drainage, the need for perioperative allogeneic transfusions and the best timing for TA administration following primary, elective, coronary artery bypass grafting (CABG) in patients with a low baseline risk of postoperative bleeding.

Methods: In this double-blind, prospective, placebo-controlled clinical trial in Seiedoshohada Hospital during 2011-2012, we evaluated 150 patients scheduled for elective, primary coronary revascularization. They were randomly divided into three groups. Group B received tranexamic 10 mg/kg prior to, Group A received tranexamic acid 10 mg/kg after cardiopulmonary bypass and group C received an equivalent volume of saline solution. Blood requirement and postoperative chest tube drainage were recorded.

Results: The placebo group (group C) had a greater postoperative blood loss 12 h after surgery (501±288 vs. 395±184 in group B and 353±181 mL in group A, P=0.004). The placebo group also had greater postoperative total blood loss (800±347 vs. 614±276 in group B and 577±228 mL in group A, P=0.001). There was a significant increase in allogeneic blood requirement in the placebo group (P=0.001).

Conclusion: For elective, first time coronary artery bypass surgery, a single dose of tranexamic acid before or after cardiopulmonary bypass is equally effective.

Background: Obsessive-compulsive disorders and depression have a high prevalence during pregnancy therefore, pregnant women may take clomipramine and also take other drugs or consume foods that contain caffeine. As investigations about the teratogenic effects of clomipramine and its concurrent administration with caffeine during organogenesis period are scarce, we aimed to study the teratogenicity of simultaneous administration of clomipramine and caffeine in rat fetus.
Methods: After dividing 42 pregnant rats to several case and control groups, we injected different doses of caffeine and clomipramine to the animals. All the injections were performed on the eighth until the 15th day of pregnancy. We removed the fetuses on the 17th day of pregnancy and studied the morphological features and apparent anomalies of the fetuses macroscopically.
Results: We found a significant rate of mortality, apparent anomalies, abnormal torsion, shrinkage of skin and subcutaneous bleeding in fetuses of rats receiving high doses of caffeine or a combination of caffeine and clomipramine. Statistical analysis of the data revealed a significant increase (P?0.001) in teratogenicity of high doses of caffeine and its combination with clomipramine.
Conclusion: This study implies simultaneous intake of high amounts of caffeine and clomipramine lead to teratogenicity. We recommend pregnant women to avoid uncontrolled consumption of foods that contain caffeine or drugs that contain high amounts of this substance. They should not also take clomipramine with caffeine in the first trimester of pregnancy.

Background: Autograft is the best option in nerve defects when end-to-end repair can not sufficiently preserve nerve continuity. Theoretically, if the severed nerve is reversely grafted, it may prevent axonal growth into nerve branches, and larger amounts of axons will reach the target organ and more satisfactory results will be obtained. In this study we aimed to compare conventional versus reverse nerve grafting.
Methods: This study was performed in Animal laboratory of Hazrat Fatemeh Hospital from April till August 2011. We randomly divided 40 Wistar rats into two groups. We excised 1.5 cm of the right sciatic nerve and anastomosed it conventionally between the proximal and distal ends of the nerve in rats in group A and in a reverse manner in rats in group B. The rats’ footprints were recorded in the first and 16th weeks after surgery. In week 16, the grafted nerves were removed under anesthesia for pathological examination and axon count. Subsequently, the results were compared clinically by sciatic functional index (SFI) through footprint analysis and paraclinically by axon count. A p-value smaller than 0.05 was considered statistically significant.
Results: Conventional and reverse nerve grafting no had statistically significant differences in clinical assessment in the first and 16th weeks (P=0.87) post-surgically and also no difference in paraclinical assessment in week 16 (P=0.68).
Conclusion: We had no significant clinically or para clinically differences between two approaches. It should be considered that the diameter and length of nerves and muscles in human is larger than rats, so the results of nerve repair may differ in human. We suggest a study in animal model which is anatomically more similar to human.

Thymoglobulin is a purified polyclonal immunoglobulin that has been used widely over the last decades in the prevention and treatment of rejection following renal transplantation. This immunoglobulin works against human thymocytes. Since thymoglobulin does not contain the nephrotoxic properties therefore it can be used in induction therapy especially in patients with higher risk of graft rejection such as patients who receive graft from cadavers. Recent research showed also its beneficial role in cross-match-positive transplantation, a role that is mediated through conjunction with inhibitors of terminal complement activation. This immunoglobulin has also been used for treatment of rejection following renal transplantation. Thymoglobulin can have various effects on various Immune system cells including T cells, B cells and also plasma cells. Thymoglobulin also affects the Tcell surface antigens, natural killer-cell antigens, B cell antigens, plasma cell antigens, adhesion molecules and chemokine receptors. Diverse effects of thymoglobulin on the immune system includes: T cell depletion, induce apoptosis in B cell lineage and interference with dendritic cell functional properties. Thymoglobulin can cause acute complications, delayed complications as well as infectious complications. Acute reaction events includes: anaphylaxis, fever, chills, dyspnea, nausea, vomiting and diarrhea. Thymoglobulin also induces cytokine release syndrome manifested by high grade fevers and chills and treated by steroid therapy. Delayed reactions events usually present as serum sickness and infections. Infectious complications are more important and include cytomegalovirus (CMV) infection, sepsis, candidiasis, herpes simplex and urinary infections. Thymoglobulin can also induce cytokine release syndrome. It has been thought that thymoglobulin increases the risk of post-transplant lymphoproliferative disorder (PTLD), however, debate still exists whether such an association is present when lower dosing regimens are used. In this review, we aimed to present first a brief history of thymoglobulin development and its mechanism of action and then assess the most recent published data regarding the role of thymoglobulin in following issues: immunological tolerance, ischemia-reperfusion injury, delayed graft function, prevention and treatment of acute allograft rejection, live donor transplantation, graft and patient survival and posttrans-plant lymphoproliferative disorder. This review can help specialist in transplant domain to appropriately used thymoglobulin in transplant patients.

Background: Graft survival has been considered the major problem in reconstructive surgery. Clinical studies have helped us to understand the role of PRP in increasing skin survival. Our goal in this study was to examine the treatment effects of platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) on autologous full thickness skin graft survival in male rats.

Methods: This experimental study was performed on 36 rats of Sprague-Dawley race with weighing approximately 250 to 300 gr on May 2015 in animal laboratory of Hazrat Fatima Hospital. After anesthesia, rats were divided into 3 groups. We injected platelet-rich plasma (PRP) in the first group, platelet-rich fibrin (PRF) in the second and saline in the third group after removing the skin. Microscopic analysis was performed with camera (Canon powershot SX200, Tokyo, Japan) on days 7, 14, 21 and 28 after surgery. We used image analysis system (ImageJ, ver. 1.45) to examine necrosis and survival rate. Samples were studied with H&E staining on day 28 microscopically for histological analysis of vascular density and angiogenesis.

Results: Our findings showed the area of necrosis in animals injected with PRP on days 7 and 14, was meaningfully less than control group (P= 0.0001). There was no meaningful difference between control and PRP groups (P> 0.05). The area of necrosis in animals injected with PRF did not have any significant difference with control group from beginning to 21st day (P< 0.0001). there was no meaningful difference in vascular density between control and PRP group, whereas in animals injected with PRF the vascular density was significantly less than control group (P= 0.002).

Conclusion: According to our results in this study, we can conclude that using autologous PRP can enhance the process of healing soft tissue injury and be affective at increasing graft survival. This method is suggested to be conducted for patients highly at risk of graft loss and also for those who are in need of early treatments.

Background: Cartilage grafting is used in rhinoplasty and reconstructive surgeries. Autologous rib and nasal septum cartilage (auto graft) is the preferred source of graft material in rhinoplasty, however, homologous cartilage (allograft) has been extensively used to correct the nasal framework in nasal deformities. Autologous cartilage graft usage is restricted with complication of operation and limiting availability of tissue for extensive deformities. Alternatively, preserved costal cartilage allograft represents a readily available and easily contoured material. The current study was a formal systematic review of complications associated with autologous versus homologous cartilage grafting in rhinoplasty patients.

Methods: In this cohort retrospective study, a total of 124 patients undergone primary or revision rhinoplasty using homologous or autologus grafts with postoperative follow-up ranging from 6 to 60 months were studied. The types of grafts and complications related to the grafts were evaluated. This included evaluation for warping, infection, resorption, mobility and fracture.

Results: The total complications related to the cartilage grafts were 7 cases, which included 1 warped in auto graft group, three cases of graft displacement (two in allograft group and one in auto graft group) and three fractures in allograft group. No infection and resorption was recorded. Complication rate (confidence interval 0.95) in autologous and homologous group were 1.25(0.4-3.88) and 2.08(0.78-5.55) in 1000 months follow up. There was no statistically significant difference between autologous and homologous group complications. Onset of complication in autologous and homologous group were 51.23(49.27-53.19) and 58.7(54.51-62.91) month respectively (P=0.81).

Conclusion: The allograft cartilage has the advantage of avoiding donor-site scar. Moreover, it provides the same benefits as autologous costal cartilage with comparable complication rate. Therefore, it can be a reliable alternative material for rhinoplasty surgeries. A longer follow-up may be necessary to confirm the structural stability of the allograft cartilage grafts.

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.

Background: The use of pesticides is essential to pests control in horticultural crops for the production of an adequate food supply for an increasing world population and for the control of insect-borne diseases. Therefore, pesticide residues in environment and food have a negative impact on the health of living organisms especially human. In this regard, the present study was carried out for assessment and comparing of propineb residues in Iranian tea and imported consumed tea.

Methods: In this experimental study, according to the Cochran's sample size formula, 12 samples from 4 brand of imported tea and 6 samples from 2 brand of Iranian tea were collected from market basket of Hamadan City in 2015. After preparation and processing the samples in the laboratory, propineb residues in samples determined using a spectrophotometric method to a wavelength range of 400 nm in 5 replications. Recovery tests were carried out using pesticide-free samples. Also, all statistical analyses were performed using the SPSS statistical package version 19 (Chicago, IL, USA).

Results: The mean concentration of propineb residues in Iranian tea samples were 1.60±0.27 µg/kg and in imported tea samples were 0.78±0.63 µg/kg, respectively and lower than the maximum residue limit (MRL) provided by the European Commission (100 ppb). The results of the comparison of the mean concentration of pesticide residues between Iranian and imported tea samples revealed no statistically significant differences between all the samples together (P=0.17).

Conclusion: The results showed that the residues of propineb were found in all analyzed tea specimens. But according to the mean concentrations of propineb residues in tea samples marketed in Hamedan City were lower than MRL, therefore, no adverse health effects have been established from controlled consuming of tea. Also, concerning increased use of agricultural inputs especially kind of pesticides for pest management by farmers, regular periodic monitoring of chemical pollutants content in Iranian and imported tea are recommended for food safety.

Background: Protective antigen of anthrax toxin, after touching the cell receptors, plays an important role in the pathogenesis of toxin. The purpose of this study was to investigate the interaction of anthrax toxin protective antigen and four great combination propolis included caffeic acid, benzyl caffeate, cinnamic acid and kaempferol using the softwares and bioinformatics web servers.

Background: Acute and chronic wound healing has always been problematic. Stem cells with or without the scaffold carrying these cells have been proposed as new methods in the treatment of wounds. In this case study we have tried to examine the effect of scaffold made of polyether sulfone (PES) alone, with stem cells and along with stem cell and growth factor on wound healing in rats.

Methods: This experimental study was conducted in Animal Laboratory of Hazrat Fatemeh Hospital in 2012. In this study, 48 rats were randomly divided into four groups. A wound created on the back of each rat at the size of 3×3 cm. The surface of the wound in the first group is covered with PES seeded with adipose-derived stem cell (ASC) and growth factor (GF), in the second group with polyether Sulfone seeded with ASC, in the third group only with PEWS, and in the fourth group (control) with Vaseline gauze. On 20^th and 35^th days, the surface of the wound was assessed by photography in order to understand the process of healing. In addition, on days 20 and 45, the histopathology characteristics of the samples were studied with a biopsy of the wounds.