Type 2 diabetes mellitus stimulated pulmonary vascular inflammation and exosome biogenesis in rats.

It has been shown that type 2 Diabetes Mellitus (T2DM) changes the paracrine activity of several cell types. Whether the biogenesis of exosomes is changed during diabetic conditions is the subject of debate. Here, we investigated the effect of T2M on exosome biogenesis in rat pulmonary tissue. Rats received a high-fat diet regime and a single low dose of Streptozocin to mimic the T2DM-like condition. A total of 8 weeks after induction of T2DM, rats were subjected to several analyses. Besides histological examination, vascular cell adhesion molecule 1 (VCAM-1) levels were detected using immunohistochemistry (IHC) staining. Transcription of several genes such as IL-1ß, Alix, and Rab27b was calculated by real-time polymerase chain reaction assay. Using western blot analysis, intracellular CD63 levels were measured. The morphology and exosome secretion activity were assessed using acetylcholinesterase (AChE) assay and scanning electron microscopy, respectively. Histological results exhibited a moderate-to-high rate of interstitial pneumonia with emphysematous changes. IHC staining showed an increased VCAM-1 expression in the diabetic lungs compared with the normal conditions (p < .05). Likewise, we found the induction of IL-1ß, and exosome-related genes Alix and Rab27b under diabetic conditions compared with the control group (p < .05). Along with these changes, protein levels of CD63 and AChE activity were induced upon the initiation of T2DM, indicating accelerated exosome biogenesis. Taken together, current data indicated the induction of exosome biogenesis in rat pulmonary tissue affected by T2DM. It seems that the induction of inflammatory niche is touted as a stimulatory factor to accelerate exosome secretion.

Thymoquinone Potentiates Methotrexate Mediated-Apoptosis in Saos-2 Osteosarcoma Cell Line.

INTRODUCTION: Recently, various studies have concentrated on the therapeutic potential of thymoquinone (TQ), a natural polyphenol, in various human malignancies, including osteosarcoma. However, the underlying mechanisms in TQ-mediated anti-cancer effects are not yet fully understood. Therefore, the present study investigated the effect of TQ on methotrexate (MTX)-induced apoptosis in Saos-2 cells. METHODS: Saos-2 cells were treated with MTX, TQ, and a combination of both, and cell viability was assessed by MTT assay. mRNA expression of apoptotic markers, including Bax, Bcl-2, and caspase-3, was assessed using quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: MTX resulted in significant inhibition of cell proliferation in a dose-dependent manner. The combination of TQ and MTX inhibited proliferation compared to single treatments (P<0.05). TQ also induced apoptosis by regulating pro-apoptotic markers including Bax and caspase-3 and reducing anti-apoptotic mediators including Bcl-2. In addition, TQ increased MTX-induced apoptosis in Saos-2 cells. CONCLUSION: The findings of the present study highlight new insights into understanding the role of TQ as a potential therapeutic agent in osteosarcoma by increasing MTX-induced apoptosis.

Melatonin Increases the Sensitivity of Osteosarcoma Cells to Chemotherapy Drug Cisplatin.

Chemotherapy, which is one of the common treatments for osteosarcoma (OS), has many side effects and in some cases has low effectiveness due to chemoresistance, hence it is vital to study new therapies for OS. In this regard, we combined melatonin with cisplatin and evaluate their effect on MG63 OS cells. Since melatonin has anti-cancer properties, we hypothesized that its combination with cisplatin could increase the effectiveness of cisplatin. Firstly, MTT assay was used to evaluate the cell viability and cytotoxicity of cisplatin on MG63 cells and the results showed that melatonin in combination with cisplatin increases the sensitivity of MG63 cells to cisplatin. In addition, qRT-PCR results showed that the expressions of miR-181 and P53, CYLD, CBX7 and BCL2 genes change in MG63 cells after treatment with the combination of cisplatin and melatonin, so that the expression of P53, CYLD and CBX7 increased and the expression of BCL2 and miR-181b decreases significantly. Furthermore, analysis of Annexin V/FITC assay data revealed that the rate of apoptosis in MG63 OS cell line remarkably promoted after treated with cisplatin and melatonin combination. As a result, our findings show that melatonin in combination with cisplatin increases the effectiveness of cisplatin in osteosarcoma cells and this study provides a new therapeutic approach for OS.

Thymoquinone Augments Methotrexate-Induced Apoptosis on Osteosarcoma Cells.

BACKGROUND: Osteosarcoma (OS) as the most frequent primary bone malignancy in children and adolescents has a short survival rate in advanced stages. Alternative herbal medicines with fewer side effects or the potency to protect common therapy's side effects can be helpful in combinational therapies. Herein, we aim to explore the effects of Thymoquinone (TQ) combined with Methotrexate (MTX) on Saos-2 cells apoptosis. METHODS: The effects of TQ and MTX alone or in combination on Saos-2 cell viability were measured by MTT assay. Real-time PCR was applied for the measurement of Bax, BCL-2, and caspase-9 mRNA expression. Apoptosis evaluation was conducted by flow cytometry. RESULTS: TQ improves the cytotoxic effects of MTX on Saos-2 cells proliferation at lower doses. Indeed, the IC50 of MTX decreased from 26 µM to 15 µM when it combined with TQ. TQ and MTX can induce the expression level of pro-apoptotic factors, Bax and caspase-9 while inhibiting anti-apoptotic protein BCL-2. Moreover, the combination of TQ and MTX potentiates apoptosis to 73%, compared to either TQ (48%) or MTX (53%) treated cells. CONCLUSION: The co-treatment of TQ and MTX is associated with the up-regulation of apoptotic factors and down-regulation of anti-apoptotic factors, conducting apoptosis aggravation and OS cell death.

DNA damage response signaling pathways as important targets for combination therapy and chemotherapy sensitization in osteosarcoma.

Osteosarcoma (OS) is the most common bone malignancy that occurs most often in young adults, and adolescents with a survival rate of 20% in its advanced stages. Nowadays, increasing the effectiveness of common treatments used in OS has become one of the main problems for clinicians due to cancer cells becoming resistant to chemotherapy. One of the most important mechanisms of resistance to chemotherapy is through increasing the ability of DNA repair because most chemotherapy drugs damage the DNA of cancer cells. DNA damage response (DDR) is a signal transduction pathway involved in preserving the genome stability upon exposure to endogenous and exogenous DNA-damaging factors such as chemotherapy agents. There is evidence that the suppression of DDR may reduce chemoresistance and increase the effectiveness of chemotherapy in OS. In this review, we aim to summarize these studies to better understand the role of DDR in OS chemoresistance in pursuit of overcoming the obstacles to the success of chemotherapy.

Chronic asthmatic condition modulated the onset of aging in bone marrow mesenchymal stem cells.

The emergence of an inflammatory condition such as asthma could affect the therapeutic potential of stem cells. Synopsis of previous documents yielded controversial outcomes, leading to a limitation of stem cell-based therapy in the clinical setting. This study aimed to assess the impact of asthmatic serum on the MSCs aging and dynamic growth in vitro. Rats were divided into control and asthmatic groups randomly. The asthmatic change was induced using OVA sensitization. The asthmatic structural changes are monitored by conventional Haematoxylin-Eosin staining. Thereafter, blood samples were taken and sera provided from each group. In this study, primary bone marrow mesenchymal stem cells were cultured in culture medium supplemented with normal and asthmatic serum for 7 days. The MSCs viability was examined using the MTT assay. The expression of the aging-related gene (ß-galactosidase), and stemness-related markers such as Sox2, Kfl-4 and p16INK4a were analysed by real-time PCR assay. Histological examination revealed chronic inflammatory remodelling which is identical to asthmatic changes. MTT assay showed a reduction of mesenchymal stem cell viability compared to the control group (P < .05). Real-time PCR analysis revealed a down-regulation of stemness-related markers Sox2, Kfl-4 and p16INK4a coincided with aging changes (ß-galactosidase) compared to the control group (P < .05). These data show the detrimental effect of asthmatic condition on bone marrow regenerative potential by accelerating early-stage aging in different stem cells and further progenitor cell depletion. SIGNIFICANCE OF THE STUDY: In such inflammatory conditions as asthma, the therapeutic potential of stem cells may be altered. We demonstrate that serum from asthmatic rats had the potential to reduce the viability of mesenchymal stem cells in vitro. Furthermore, we observed that the expression of the aging-related gene known ß-galactosidase was statistically increased in cells co-cultured with asthmatic serum. At the same time, expression of stemness-related markers Sox2, Kfl-4 and p16INK4a down-regulated. These results support the damaging effect of asthmatic condition on bone marrow regenerative ability by inducing early-stage aging in stem cells and additional progenitor cell reduction.

CRISPR/Cas9 novel therapeutic road for the treatment of neurodegenerative diseases.

CRISPR (clustered regularly interspaced short palindromic Repeats)/Cas9 is a new genetic editing technology that can be a beneficial method to advance gene therapy. CRISPR technology is a defense system of some bacteria against invading viruses. Genome editing based on the CRISPR/Cas9 system is an efficient and potential technology that can be a viable alternative to traditional methods. This system is a compound of a short guide RNAs (gRNAs) for identifying the target DNA sequence and Cas9 protein as nuclease for breaking and cutting of DNA. In this review, recent advances in the CRISPR/Cas9-mediated genome editing tools are presented as well as their use in gene therapy strategies for the treatment of neurological disorders including Parkinson's disease, Alzheimer's disease, and Huntington's disease.

Orthopedic injuries following the East azerbaijan earthquake.

BACKGROUND: Evaluating demographic characteristics, distribution and types of orthopedic injuries following major earthquakes may be helpful in future planning for disasters. OBJECTIVES: This study aimed to analyze data from trauma patients with extremity injury resulting from the earthquakes of East Azerbaijan, Iran. PATIENTS AND METHODS: Medical records of 686 patients admitted to Shohada hospital, Trauma Center of Tabriz University of Medical Sciences were reviewed. There were 200 patients with extremity injury assessed. Demographic characteristics and patterns of injuries in these patients were evaluated. RESULTS: In this study, there were 105 females (52.5%) and 95 males (47.5%), out of which, 6 (3%) patients with associated severe head injuries died. The most common sites of injury were lower extremities (81 patients, 41.5% of total victims) while 32 patients (16%) suffered from both upper and lower extremity injuries. Open Fractures were seen in lower extremities of 26 (13%) patients. Compartment syndrome was observed in 3 (1.5%) patients with lower limb fractures. Also, 42 (21%) patients living in Tabriz were injured while they were running away (falling down the stairs and jumping out of windows). CONCLUSIONS: Extremity injuries especially open fractures of lower limbs account for the majority of hospitalized victims. Compartment syndrome is one of the main problems that should be addressed. Our study demonstrates that people still need more education regarding earthquakes and the government should direct more attention to this issue.

Adolescence spinal epidural abscess with neurological symptoms: case report, a lesson to be re-learnt.

Epidural abscess of the spinal column is a rare condition that can be fatal if left untreated. It promptly progresses and can cause neurologic paralysis, urinary retention or cauda equina syndrome. Compromised immune system that occurs in patients with diabetes mellitus, AIDS, chronic renal failure, alcoholism, or cancer is a predisposing factor. It mostly occurs in adults. Here we would like to report a case of spontaneous pyogenic lumbar epidural abscess with neurological deficit diagnosed in a 15 year old boy. We treated this case successfully with surgical microscopic decompression and drainage.

Comparison of titanium elastic nails with traction and spica cast in treatment of children's femoral shaft fractures.

Titanium Elastic Nails (TEN) are commonly used to stabilize femoral fractures in school-aged children, but there have been few studies assessing the risk of traditional traction and application of spica cast. The aim of this study was to compare of titanium elastic nails with traction and spica cast in treatment of children's femoral shaft fractures. A group of thirty children aged 6-12 years with one-sided femoral shaft fracture were randomly allocated either to traction with spica casting group or titanium elastic nails group and were followed up to 1 year. Factors such as age, sex, time needed for walking with aids, time needed for independent walking, time needed for callus formation, time absent from school, time spent in hospital, malunion, malalignment and wound complication were recorded and compared. Fifteen patients (10 boys, 5 girls) with a mean age of 8.33 +/- 1.63 years were treated by traction and spica casting. The other 15 (9 boys, 6 girls) with a mean age of 8.73 +/- 1.53 years underwent surgery using TEN. Mean absence time from school, length of hospital stay, time needed for walking with and without help and angular deviation (varus or valgus) were significantly lower in the group treated by TEN (p-value < 0.05). Time needed for callus formation was significantly lower in spica casting group (p-value < 0.001). No statistically significant difference was observed between the two groups regarding malunion wound complications and hospital charges. The results indicated that a child in whom a femoral fracture is treated with TEN achieves recovery milestones significantly faster than a child treated with traction and spica cast.

A short term follow up comparison of genu varum corrective surgery using open and closed wedge high tibial osteotomy.

INTRODUCTION: Knee deformity associated with osteoarthritis (OA) is one of the most common complications seen in patients referred to orthopaedic surgeons. High tibial osteotomy (HTO) is an accepted method for treatment of medial knee osteoarthritis with varus deformity. The aim of this study was to compare results of osteotomy methods in patients with genu varum (GV) deformity. METHODS: In this cohort study, the sample consisted of 32 patients with genu varum deformity (42 knees) who were divided into two groups and matched according to age and gender. The patients were treated with open or closed wedge osteotomy. After surgery, they were followed-up and compared for 6 months. RESULTS: The sample consisted of 25 women (87.2%) and 7 men (21.8%). Ten patients (31.2%) presented with bilateral deformity. The incidence of complications was the same for both procedures (12.5%); this included one peroneal nerve injury following closed wedge surgery. Overall, patient satisfaction was 87.5% and 75% for the open and closed wedge methods respectively. Operative time, days to full weight bearing, and days to return to routine activities were significantly shorter for patients treated with the open wedge method (p<0.001). CONCLUSION: Open wedged HTO is associated with shorter operating time, shorter recovery time, higher patient satisfaction, and probable lower risk of neurological injury. KEY WORDS: High Tibial Osteotomy, Open Wedge Osteotomy, Closed Wedge Osteotomy.

AO tubular external fixation vs. unreamed intramedullary nailing in open grade IIIA-IIIB tibial shaft fractures: a single-center randomized clinical trial.

Although the intramedullary nailing is thought to be the method of choice for treatment of closed tibial shaft, there is ongoing debate on the optimal surgical approach in patients with open types of these fractures. In addition, choosing between the reamed and unreamed intramedullary nailing is still an issue for the orthopedic surgeons. In present study, we aimed to compare the outcome and consequences of OA tubular external fixation vs. unreamed intramedullary nailing in open grade IIIA-IIIB tibial shaft fractures. In a randomized clinical trial, 50 patients with open tibial shaft fractures (grades IIIA-IIIB) were recruited in Tabriz Shohada teaching centre in a 2-year period of time. They randomized in two equal groups underwent either AO tubular external fixation or unreamed intramedullary nailing. These 2 groups were matched for sex, age and fracture-grade. The follow-up time was one year. Union time, surgical outcome, postoperative complications and the ambulation time were compared between the two groups. Twenty five patients, 20 males and 5 females with a mean age of 30.80 +/- 5.24 years were allocated in unreamed intramedullary group and 25 other patients, 22 males and 3 females with a mean age of 28.92 +/- 8.88 years were studied in the external fixation group (p = 0.70 and 0.37, respectively). The two groups were matched for sex (p = 0.70) and age (p = 0.37). The time of union was 3, 4, 5 and 6 < or = weeks after operation in 28, 12, 32 and 28% of the cases in unreamed intramedullary group vs. 4, 12, 48 and 36% of the cases in external fixation group, respectively (p = 0.14). Post-operative infection, soft tissue injury, malunion and nonunion were documented in 16, 8, 0 and 4% of the cases in unreamed intramedullary group vs. 32, 12, 24 and 8% of the cases in external fixation group, respectively (p = 0.19, 0.50, 0.02 and 0.50, respectively). The mean ambulation time after operation was 2.92 +/- 2.43 weeks in the unreamed intramedullary nailing group vs. 2.68 +/- 2.14 weeks in the external fixation group (p = 0.71). Our results are in favor of unreamed intramedullary nailing against external fixation in treatment of open tibial shaft fractures.

Lenke and King classification systems for adolescent idiopathic scoliosis: interobserver agreement and postoperative results.

PURPOSE: The aim of this study was to investigate the interobserver agreement of the Lenke and King classifications for adolescent idiopathic scoliosis, and to compare the results of surgery performed based on classification of the scoliosis according to each of these classification systems. METHODS: The study was conducted in Shohada Hospital in Tabriz, Iran, between 2009 and 2010. First, a reliability assessment was undertaken to assess interobserver agreement of the Lenke and King classifications for adolescent idiopathic scoliosis. Second, postoperative efficacy and safety of surgery performed based on the Lenke and King classifications were compared. Kappa coefficients of agreement were calculated to assess the agreement. Outcomes were compared using bivariate tests and repeated measures analysis of variance. RESULTS: A low to moderate interobserver agreement was observed for the King classification; the Lenke classification yielded mostly high agreement coefficients. The outcome of surgery was not found to be substantially different between the two systems. CONCLUSION: Based on the results, the Lenke classification method seems advantageous. This takes into consideration the Lenke classification's priority in providing details of curvatures in different anatomical surfaces to explain precise intensity of scoliosis, that it has higher interobserver agreement scores, and also that it leads to noninferior postoperative results compared with the King classification method.

Effect of low-intensity pulsed ultrasound on fracture callus mineral density and flexural strength in rabbit tibial fresh fracture.

BACKGROUND: Low-intensity ultrasound is a biophysical intervention on a fracture repair process. However, the effect of low-intensity ultrasound therapy on fracture healing is controversial. The aim of the present study was to evaluate the effect of low-intensity pulsed ultrasound (LIPUS) therapy on the fracture healing process, including mineral density and strength of callus using a rabbit model. METHODS: A total of 30 rabbits underwent unilateral, transverse, and mid-tibia open osteotomies that were stabilized with external fixators. Then, the animals were divided into two study groups composed of 15 rabbits each: the case group (US), which were exposed to low-intensity pulsed ultrasound with 30 mW/cm(2) intensity and 1.5 MHz sine waves; and the control group (C), which underwent sham ultrasound treatment. Callus development and mineral density were evaluated using multidetector computed tomography at 2, 5, and 8 weeks, after which the animals were killed. Three-point bending tests of both healed and intact bones were assessed and compared. RESULTS: The results demonstrated that the callus mineral density in the US group was higher than in the C group (1202.20 +/- 81.30 vs. 940.66 +/- 151.58 HU; P = 0.001) at the end of the 8th week. The mean recorded three-point bending test score of healed bones in the US group was not significantly different from that of the C group (359.35 +/- 173.39 vs. 311.02 +/- 80.58 N; P = 0.114). CONCLUSIONS: The present study showed that low-intensity pulsed ultrasound enhanced callus mineral density with an insignificant increase in the strength of the fractured bone.

Effect of low-level laser therapy on the fracture healing process.

Low-level laser therapy (LLLT) is a biophysical form of intervention in the fracture-repair process, which, through several mechanisms, accelerates the healing of fractures and enhances callus formation. The effect of laser on fracture healing is controversial. Some authors affirm that LLLT can accelerate bone formation by increasing osteoblastic activity. The objective of our study was to evaluate the effect of laser therapy on fracture healing. Thirty rabbits were subjected to tibial bone open osteotomies that were stabilized with external fixators. The animals were divided into two study groups: laser group and control group. Callus development and bone mineral density were quantitatively evaluated by CT; the animals were then killed and the fractures were assessed for biomechanical properties. The results demonstrated that the increasing rate of bone mineral density was higher in the laser (L) group than in the control (C) group. CT at 5 weeks revealed a mean callus density of 297 Hounsfield units (HU) for the control group and 691 HU for the L group, which was statistically significant (P = 0.001). In the L group, the mean recorded fracture tension was 190.5 N and 359.3 N for healed and intact bones, respectively, which was statistically significant (P < 0.001). The result of the study showed that the use of laser could enhance callus development in the early stage of the healing process, with doubtful improvement in biomechanical properties of the healing bone; therefore, laser therapy may be recommended as an additional treatment in non-union fractures in humans.

Comparison between ender and unreamed interlocking nails in tibial shaft fractures.

OBJECTIVE: To compare ender nails and unreamed interlocking nails in a randomized clinical trial study. METHODS: From March 2001 to March 2003, 131 patients with tibial fractures of the central two thirds of the tibia were admitted to the emergency ward of Shohada Hospital, Tabriz, Iran and were randomized to either an ender tibial nail or an undreamed interlocking tibial nail as their primary treatment. Age, gender, types of fracture, and surgical outcomes were recorded at the time of surgery, during hospital stay, and during the course of 9 months follow-up. RESULTS: Sixty-seven patients underwent surgery with interlocking nails and 64 with ender nails. No statistically significant difference was found between the 2 groups concerning mal-union, delayed union, and device failure. Durations of hospital stay, and time needed to achieve union was significantly higher in the unreamed interlocking group. Non-union and infection rate were higher in the ender group (p<0.05). CONCLUSION: The results of our study support those of other studies that indicate the choice between the 2 methods of unreamed nailing is mainly controversial, depending on the priorities of the patient and surgeon.