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Inhibiting exosomal MIC-A and MIC-B shedding of cancer cells to overcome immune escape: new insight of approved drugs

Abstract: Our knowledge of the role of innate immunity in protecting against cancers has expanded greatly in recent years. An early focus was on the adoptive transfer of natural killer (NK) cells and, although this approach has demonstrated promising results in many patients, a few limitations including immune escape of tumors from cytotoxic killing by NK cells have caused treatment failures. Downregulation of the expression of activating ligands on the surface of cancer cells and prevention of the activity of soluble factors are among the mechanisms employed by cancer cells to overcome NK-mediated immunity. It has become evident that a class of small membranous structures of endosomal origin known as exosomes play a key role in regulating the local tumor microenvironment. Here we hypothesize that exosome secretion by cancer cells, which is greater than that of normal cells, is an important escape mechanism employed by cancer cells. Interruption of exosome release by various inhibitory agents in combination with the adoptive transfer of NK cells may overcome, at least in part, the treatment failures that occur with adoptive NK cell transfer. In this regard, repositioning of approved drugs with previously shown effects on exosome release may be a good strategy to bypass the safety issues of newly identified agents and will also dramatically reduce the huge costs of drug approval process. © 2019, Springer Nature Switzerland AG.

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Diagnostic accuracy of interferon (IFN)-γ inducible protein 10 (IP-10) as a biomarker for the discrimination of active and latent tuberculosis

Abstract:To assess the potency of Interferon (IFN)-γ inducible protein 10 (IP-10) stimulated by recombinant PE35 and PPE68 as a biomarker in differentiating between active and latent tuberculosis. Patients with active pulmonary TB (PTB) (n = 30), latent TB infection (LTBI) (n = 29), and BCG-vaccinated healthy controls (HCs) (n = 30) were enrolled and blood samples were taken from them. The diagnostic performance of IP-10 was evaluated by the Receiver operator characteristic (ROC) curve and the area under the curve (AUC) and their 95% confidence intervals (CI) were calculated. The median IP-10 concentrations following stimulation with recombinant PE35 and PPE68 were significantly higher in TB-infected group (both PTB and LTBI) compared with HCs (P < 0.05). It was also significantly higher in PTB patients compared with individuals with LTBI (P < 0.05). The discriminatory performance of IP-10 following stimulation with recombinant PE35 and PPE68 (assessed by AUC) between TB patients and HCs were similar (AUC: 0.79 [95% CI 0.68–0.89] and 0.79 [95% CI 0.69–0.89], respectively). AUCs of IP-10 following stimulation with recombinant PE35 and PPE68 for distinguishing between PTB and LTBI groups were 0.63 (95% CI 0.47–0.79) and 0.61 (0.45–0.77), respectively. Under the selected cut-off values, the sensitivity and specificity of IP-10 for distinguishing of TB-infected and HCs after stimulation with recombinant PE35 was 74.5% and 73%, respectively and after stimulation with recombinant PPE68 were 76.5% and 63%, respectively. Moreover, the sensitivity and specificity of IP-10 for differentiating of PTB and LTBI following stimulation with recombinant PE35 and PPE68 were 770 pg/ml (sensitivity: 63%; specificity: 62%) and 502 pg/ml (sensitivity: 80%; specificity: 52%), respectively. IP-10 stimulated by recombinant PE35 and PPE68 is a promising biomarker for TB diagnosis. However, it doesn’t have desirable sensitivity and specificity in distinguishing between PTB and LTBI. © 2019, Springer Nature B.V.

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Histomorphometric and immunohistochemical evaluation of angiogenesis in ischemia by tissue engineering in rats: Role of mast cells

Abstract:The aim of this study was to find a proper method for improvement of ischemic condition in the rat hind limb and also to observe the efficacy of cell engraftment with alginate/gelatin three-dimensional scaffolds. Eighteen male Wistar rats weighing 200 to 250 g were randomly divided into three groups (n = 6) including a) ischemia group; in which femoral artery was removed after ligation at the distance of 5 mm, b) scaffold group; in which hydrogel scaffold was added to the site of transected femoral artery and c) test group; in which in addition to hydrogel scaffold, mast cells (MCs) were also added (1 × 106 cells). Analysis of capillary density, artery diameter, histomorphometric parameters and immunohistochemistry in transected location were done on day 14 after femoral artery transection. The average number of blood capillary was significantly higher in the test group than other groups. Also, the average number of medium and large blood vessels was significantly higher in the test group compared to ischemia and scaffold groups. Application of MCs through the use of hydrogel scaffolds (alginate/gelatin) can be considered as a new approach in the application of stem cells for therapeutic angiogenesis under ischemic conditions which can improve the angiogenesis process in patients with peripheral artery diseases. © 2019 Urmia University. All rights reserved.

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Correction: Exhaled nitric oxide is not a biomarker for idiopathic pulmonary arterial hypertension or for treatment efficacy (BMC Pulmonary Medicine DOI: 10.1186/s12890-019-0954-z)

Abstract:Following publication of the original article [1], the authors flagged that name of the author Batoul Khoundabi had been provided with an incorrect spelling: Batoutl was given in place of Batoul. This typo has now been corrected in the original article and the corrected version is provided with this correction. © 2019 The Author(s). Reference:.

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Exhaled nitric oxide is not a biomarker for idiopathic pulmonary arterial hypertension or for treatment efficacy

Abstract:Background: Idiopathic pulmonary arterial hypertension (IPAH) is a fatal illness. Despite many improvements in the treatment of these patients, there is no unique prognostic variable available to track these patients. The aim of this study was to evaluate the association between fractional exhaled nitric oxide (FeNO) levels, as a noninvasive biomarker, with disease severity and treatment outcome. Methods: Thirty-six patients (29 women and 7 men, mean age 38.4 ± 11.3 years) with IPAH referred to the outpatients clinic of Masih Daneshvari Hospital, Tehran, Iran, were enrolled into this pilot observational study. Echocardiography, six-minute walking test (6MWT), FeNO, brain natriuretic peptide (BNP) levels and the functional class of patients was assessed before patients started treatment. Assessments were repeated after three months. 30 healthy non-IPAH subjects were recruited as control subjects. Results: There was no significant difference in FeNO levels at baseline between patients with IPAH and subjects in the control group. There was also no significant increase in FeNO levels during the three months of treatment and levels did not correlate with other disease measures. In contrast, other markers of disease severity were correlated with treatment effect over the three months. Conclusion: FeNO levels are a poor non-invasive measure of IPAH severity and of treatment response in patients in this pilot study. © 2019 The Author(s).

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Characterization of Clostridioides difficile isolates recovered from hospitalized patients and the hospitals environment and air: A multicenter study

Abstract:In healthcare settings, contamination of environment with toxigenic and hypervirulent Clostridioides difficile strains is a serious concern. Here, we assessed whether patients with C. difficile have a role to play in the dissemination of C. difficile in our settings or other sources are implicated in its circulation. A total of 700 fecal specimens and 1435 environmental samples from surfaces, equipment and air of rooms occupied by patients suspected of C. difficile infection were taken from 4 tertiary hospitals in Tehran, Iran between April 2016 and August 2017. Antibiotic susceptibility testing and detection of resistance genes were performed for the environmental isolates. The clinical and environmental isolates of C. difficile were subjected to Pulsed Field Gel Electrophoresis (PFGE) analysis. Forty three (6.14%) and 2 (0.13%) isolates of C. difficile were recovered from the clinical and environmental samples, respectively. In the clinical settings, 2 patients were suspected of recurrent C. difficile infection. Thirty distinct pulsotypes were found among the C. difficile isolates including 28 singletons and 2 common types. One of the two environmental isolates was isolated from floor in the Medical ward, of pulsotype/ribotype/toxinotype PT10/New ribotype/toxinotype V, harbored cdtA/B and tcdC-A, and resistant to ciprofloxacin. The other one was isolated from air of a room in ICU, assigned to PT11/RT001/toxinotype 0, belonged to tcdC-sc3 genotypes and resistant to metronidazole. The environmental isolates did not generate amplicons in PCR assays targeting vanA and nim genes. This study provided evidence for dissemination of genetically diverse strains of C. difficile in hospitalized patients, presence of C. difficile in hospital air, existence of binary toxin positive/antibiotic-resistant isolate on the floor and intra-hospital dissemination of this pathogen. © 2019 Elsevier Ltd

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Does Neutrophil Phenotype Predict the Survival of Trauma Patients?

Abstract:According to the World Health Organization (WHO), trauma is responsible for 10% of deaths and 16% of disabilities worldwide. This is considerably higher than those for malaria, tuberculosis, and HIV/AIDS combined. While the human suffering and death caused by injury is well-recognized, injury has a significant medical care cost. Better prediction of the state of trauma patients in the days immediately after trauma may reduce costs. Traumatic injuries to multiple organs can cause dysfunction in all systems of the body especially the immune system placing patients at high risk of infections and inflammatory complications which are often fatal. Neutrophils are the most abundant leukocyte in the human circulation and are crucial for the prevention of microbial disease. Significant changes in neutrophil functions such as enhanced chemotaxis, Neutrophil extracellular trap (NET)-induced cell death (NETosis), and phagocytosis occur early after injury followed by prolonged functional defects such as phagocytosis, killing mechanisms, and receptor expression. Analysis of these changes may improve the prediction of the patients condition over time. We provide a comprehensive and up-to-date review of the literature investigating the effect of trauma on neutrophil phenotype with an underlying goal of using this knowledge to examine the predictive potential of neutrophil alterations on secondary complications in patients with traumatic injuries. We conclude that alterations in neutrophil surface markers and functions may be potential biomarkers that predict the outcome of trauma patients. © Copyright © 2019 Mortaz, Zadian, Shahir, Folkerts, Garssen, Mumby and Adcock.

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A bioinformatics analysis of exosomal microRNAs released following mycobacterial infection

Background: Tuberculosis (TB) still remains a major health threat worldwide. The current TB diagnostics are suboptimal, and there is a high clinical need for identifying novel biomarkers of disease prevalence. Circulating exosomes have been currently attractive as novel biomarkers in a wide range of pathological conditions. Methods: In this study, we performed bioinformatics analysis on the downstream targets of a dysregulated microRNA (miRNA) cluster induced by Bacillus Calmette-Guerin infection of human macrophages to provide greater understanding of their potential roles in disease pathogenesis. Results: Our analysis demonstrated that these dysregulated miRNAs have central roles in the host metabolic and energy pathways. Conclusion: This suggests that the host miRNA network is perturbed by Mycobacterium to re-patterning host metabolism machinery to favor its intracellular survival. The dysregulated miRNAs can be delivered to local and distal cells by exosomes and thereby modulate their function. © 2019 International Journal of Mycobacteriology | Published by Wolters Kluwer-Medknow.

We evaluated the potential role of IL-2 in whole blood stimulated with Mycobacterium tuberculosis-specific antigens in the QuantiFERON-TB Gold In Tube (QFT-G-IT) for the discrimination of active and latent tuberculosis. In this study, patients were recruited from the infectious diseases ward at the Masih Daneshvari Hospital, National Research Institute of Tuberculosis and Lung Diseases (NRITD), affiliated to Shahid Beheshti University of Medical Sciences, Tehran, Iran. Individuals were classified as having active TB when the diagnosis was confirmed by positive M. tuberculosis culture from sputum specimens and positive response to QFT-G-IT test. LTBI was defined as a positive response to QFT-G-IT in the absence of symptomatic, microbiological, or radiological evidence of active tuberculosis. Healthy controls were BCG vaccinated individuals with no known exposure to M. tuberculosis and a negative response to the QFT-G-IT. We do not include in the study individuals who had positive human immunodeficiency virus screening result. The study received approval from the Ethical Committee of Tehran University of medical sciences.

In this study, thirty patients with active TB and 30 cases with LTBI had positive QFT-G-IT test and all of the controls (N = 30) had negative QFT-G-IT result. After 72-h of stimulation by antigens from the QFT-G-IT assay, IL-2 secretion was quantitated in supernatants by using ELISA (Mabtech AB, Sweden). All the plasma specimens were tested in duplicate and expressed in pg/ml. The differences in levels of IL-2 among groups were analyzed using non-parametric analysis of variance with the Kruskall–Wallis test. As shown in Fig. 1, LTBI group induced significantly greater production of IL-2 than patients with active TB infection (P value = 0.019, Kruskal–Wallis test). The discrimination performance (assessed by the area under ROC curve) between LTBI and patients with active TB was 0.816 (95%CI: 0.72–0.97) (Fig. 2). Cut-offs were estimated at various sensitivities and specificities and at the maximum Youdens index (YI), i.e. sensitivity specificity–1. Maximum discrimination was reached at a cut-off of 13.9 pg/mL for IL-2 following stimulation with 82% sensitivity, 86% specificity, 85% PPV and 83% NPV.

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Successful control of exacerbation of Allergic Bronchopulmonary Aspergillosis due to Aspergillus terreus in a cystic fibrosis patient with short-term adjunctive therapy with voriconazole: A case report

Abstract:Background: A 12-year-old boy with cystic fibrosis (CF) and a history of glucocorticoid-dependent allergic bronchopulmonary aspergillosis (ABPA) was referred to our hospital. The ABPA was diagnosed when he was 8 years old and he had been treated with several course of oral glucocorticoids for recurrent exacerbations. He was readmitted when aged 12 with a history of worsening shortness of breath and chest tightness. A recurrence of ABPA was diagnosed based on eosinophilia and elevation of Aspergillusspecific IgE and IgG, and total IgE. Thoracic high-resolution computed tomography (HRCT) showed central bronchiectasis with parenchymal infiltrates. The treatment started with itraconazole and oral corticosteroid. After 2 months of treatment, he was re-admitted to the hospital due to a progressive worsening of respiratory symptoms. Chest HRCT revealed the a sub segmental atelectasis in the left lung. Microscopic examination of sputum and BAL samples demonstrated septate hyphae consistent with Aspergillus species. Sputum and BAL culture yielded Aspergillus ochraceus and Aspergillus terreus, which were both sensitive to itraconazole and voriconazole. The treatment was switched to voriconazole and the patient showed significant clinical, serological and mycological improvement after three months. This case shows that voriconazole may be used as an alternative for treatment of ABPA due to Aspergillus terreus. © 2019 Elsevier Masson SAS

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Common Infections and Target Organs Associated with Chronic Granulomatous Disease in Iran

Abstract:Recurrent severe bacterial and fungal infections are characteristic features of the rare genetic immunodeficiency disorder chronic granulomatous disease (CGD). The disease usually manifests within the first years of life with an incidence of 1 in approximately 200,000 live births. The incidence is higher in Iran and Morocco where it reaches 1.5 per 100,000 live births. Mutations have been described in the 5 subunits of NADPH oxidase, mostly in gp91 phox and p47 phox, with fewer mutations reported in p67 phox, p22 phox, and p40 phox. These mutations cause loss of superoxide production in phagocytic cells. CYBB, the gene encoding the large gp91 phox subunit of the transmembrane component cytochrome b 558 of the NADPH oxidase complex, is localized on the X-chromosome. Genetic defects in CYBB are responsible for the disease in the majority of male CGD patients. CGD is associated with the development of granulomatous reactions in the skin, lungs, bones, and lymph nodes, and chronic infections may be seen in the liver, gastrointestinal tract, brain, and eyes. There is usually a history of repeated infections, including inflammation of the lymph glands, skin infections, and pneumonia. There may also be a persistent runny nose, inflammation of the skin, and inflammation of the mucous membranes of the mouth. Gastrointestinal problems can also occur, including diarrhea, abdominal pain, and perianal abscesses. Infection of the bones, brain abscesses, obstruction of the genitourinary tract and/or gastrointestinal tract due to the formation of granulomatous tissue, and delayed growth are also symptomatic of CGD. The prevention of infectious complications in patients with CGD involves targeted prophylaxis against opportunistic microorganisms such as Staphylococcus aureus, Klebsiella spp., Salmonella spp. and Aspergillus spp. In this review, we provide an update on organ involvement and the association with specific isolated microorganisms in CGD patients. © 2019 S. Karger AG, Basel. All rights reserved.

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Procalcitonin and proinflammatory cytokines in early diagnosis of bacterial infections after bronchoscopy

Abstract:BACKGROUND: Fiberoptic bronchoscopy (FOB) guided bronchoalveolar lavage (BAL) remains as the chief diagnostic tool in respiratory disorders. 1.2-16% of patients frequently experience fever after bronchoscopy. To exclude the need for multiple antibiotic prescribing in patients with post-bronchoscopy fever, the presence of the self-limiting inflammatory responses should be excluded. AIM: The current study was conducted to test the serum of patients undergoing bronchoscopy for some proinflammatory cytokines including Tumor Necrosis Factor-alpha (TNF-?), Interleukin-1beta (IL-1β), Interleukin-8 (IL-8) and Interleukin-6 (IL-6) and the value of Procalcitonin (PCT). MATERIAL AND METHODS: Current case-control study was conducted at the National Research Institute of Tuberculosis and Lung Disease in Iran. Nineteen patients (48.72%) that attended with a reasonable sign for a diagnostic bronchoscopy from January 2016 to December 2017 were included in the case group. The control group consisted of 20 patients who underwent a simple bronchoscopy and without FOB-BAL. The laboratory findings for PCT concentrations and cytokine levels in the three serum samples (before FOB-BAL (t0), after 6 hr. (t1), and at 24 hr. past (t2) FOB-BAL) were compared between two groups. RESULTS: The frequency of post-bronchoscopy fever was 5.12, and the prevalence of post-bronchoscopy infectious fever was 2.56%. PCT level was considerably higher in the patient with a confirmed bacterial infection when compared to other participants (p-value < 0. 05). Interestingly, IL-8 level in the bacterial infection proven fever patient was higher than in other patients (p < 0.001). IL-8 levels displayed a specificity of 72.7% and a sensitivity of 100%, at the threshold point of 5.820 pg/ml. PCT levels had a specificity of 84% and a sensitivity of 81%, at the threshold point of 0.5 ng/ml. CONCLUSION: The present findings show that in patients with fever after bronchoscopy, PCT levels and IL-8 levels are valuable indicators for antibiotic therapy, proving adequate proof for bacterial infection. The current findings also illustrate that to monitor the serum levels of PCT and proinflammatory cytokines in the patients undergoing FOB-BAL, the best time is the 24-hour postoperative bronchoscopy. © 2019 Mohsen Farrokhpour, Arda Kiani, Esmaeil Mortaz, Kimia Taghavi, Amir Masoud Farahbod, Atefeh Fakharian, Mehdi Kazempour-Dizaji, Atefeh Abedini.

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Phenotypical and functional evaluation of dendritic cells after exosomal delivery of miRNA-155

Abstract:Aims: The clinical efficiency of dendritic cell (DC) therapy needs to be improved. Exosomes, as membrane nano-vesicles, carry bio-macromolecules and play essential roles in intercellular crosstalk. Here, it is proposed that tumor cell-derived exosomes could function as vehicles to deliver exogenous miRNA-155 into DCs, for simultaneous miRNA delivery and antigen priming of DCs. Following optimization of the miRNA-155 delivery, the effect of exogenous miRNA-155 overexpression on DCs is evaluated. Main methods: For this purpose, exogenous miRNA-155 was electroporated with various voltages (0.100, 0.200, and 0.300 kV) into tumor cell-derived exosomes with various concentrations, and then DCs were treated with miRNA-155 loaded exosomes. To assess the effect of miRNA-155 loaded exosomes on DCs, the expression levels of IL12p70, IFN-γ and IL10 in culture supernatants were measured by ELISA. Then, the expression profiles of DC surface markers, including CD11C, MHCII (I/A-I/E), CD86, CD40, and CD83 were investigated by flow cytometry. Key findings: Concerning the results, exogenous miRNA-155 can be successfully inserted into tumor cell derived exosomes. Loading conditions for tumor cell-derived exosomes were enhanced for utilization as vehicles to deliver miRNA-155 into DCs. Analysis of the surface molecule revealed that miRNA-155 can increase the expression levels of MHCII (I/A-I/E), CD86, CD40, and CD83. ELISA analysis indicates that miRNA-155 can significantly increase, the levels of IL12p70, IFN-γ and IL10. Significance: Finally, it can be stated that miRNA-155 could be a candidate for dendritic cell maturation. This method can be applied in the modification of target cells in in vitro studies. © 2019

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Clinical Presentation of Nontuberculous Mycobacteria Using Radiological and CT Scan Imagining

Abstract:In this chapter, chest X-ray and computed tomography findings are used to differentiate between nontuberculous mycobacteria (NTM) and tuberculosis infection. Common and recognized patterns of NTM infection include cavitary diseases, bronchiectasis, and pulmonary infections in AIDS patients. Less commonly, the nodular pattern, a mass mimicking malignancy and of hypersensitivity pneumonitis can be seen. Any diagnosis requires a combination of laboratory, clinical and radiological findings. © 2019 Elsevier Inc. All rights reserved.

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Serum interleukin-27 level in different clinical stages of lung cancer

Abstract:BACKGROUND: Advanced lung cancer is indicated with rapid disease development. Interleukin 27 (IL-27) is regarded as a cytokine with anti-tumour activities. AIM: Since, the impact of type of lung cancer on the level of IL-27 in patient’s serum has not yet been investigated; current study evaluated the clinical stages according to American Joint Committee on Cancer (AJCC) criteria, Tumor-Node-Metastasis (TNM) stage and the lung cancer spread (localized or widespread) and its correlation with serum IL-27. MATERIAL AND METHODS: Thirty patients with confirmed histopathological lung cancer and 30 cancer-free healthy individuals as the control group were included in the current study. Patients group were assigned to either small cell lung cancer group (SCLC) or non-small cell lung cancer (NSCLC) according to the clinical features and the results of lung biopsy specimens. Level of IL-27 was quantified with enzyme-linked immunosorbent assay (ELISA) test in serum samples. RESULTS: A significant increase in serum IL-27 level was noticed in individuals with lung cancer in comparison with the control group. The level of serum IL-27 in the NSCL squamous carcinoma (NSCLC-Sc) type was significantly greater than in the NSCLC adenocarcinoma (NSCLC-Ad) type, and in both groups, this variable was more than the control group. The serum IL-27 content level was greater in stage III versus stage IV. CONCLUSION: The current research confirmed the existence of the anti-tumour components in patients with NSCLC. IL-27 can be utilised in diagnosis and screening in early stages of lung cancer along with the management of patients. Different levels of IL-27 in different types of lung cancers in the current study can lead to design more comprehensive studies in the future. © 2019 Akbar Soleimani Babadi, Arda Kiani, Esmaeil Mortaz, Kimia Taghavi, Adnan Khosravi, Majid Marjani, Sharareh Seifi, Habib Emami, Atefeh Abedini.

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Toothpick aspiration induces massive hemoptysis: A case report

Abstract:Background: Massive hemoptysis refers to bleeding from the sputum exceeding 100 ml/day. This condition is known to have a poor prognosis. Although foreign body aspiration is not as common as other risk factors, it may result in massive hemoptysis. In the current study, we presented a case of massive hemoptysis due to the aspiration of a toothpick. Case Presentation: The patient was a 49-year-old woman who was primarily suspected of having tuberculosis. After observing blood in the sputum, interventions, including chest computed tomography (CT) scan and conservative management, were performed. The CT scan showed no malignancy, and paraclinical investigations were negative. However, hemoptysis was progressing into an acute phase; therefore, a surgical intervention was performed for the patient. After the surgery, the cause of the lesion was found to be a toothpick. The patient was under intensive care after surgery and was discharged from the hospital in a good general condition. The morphological evaluation of the lesion showed a bronchial wall with ulceration, besides granulation tissue formation, hematoma, and fibrinoid necrosis due to foreign body aspiration into the lung, resulting in inflammatory reactions. Conclusion: In this case report, foreign body aspiration resulted in massive hemoptysis. Our primary attempts to diagnose the cause of lesion were unsuccessful, and surgery was performed due to the life-threatening condition of the patient. Overall, unexplained hemoptysis may occur following a serious accident due to foreign body aspiration. ©2019 NRITLD, National Research Institute of Tuberculosis and Lung Disease, Iran.

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Evaluation of Silymarin for management of anti-tuberculosis drug induced liver injury: A randomized clinical trial

Abstract:Aim: This study was performed to evaluate the potential efficacy of silymarin in the management of anti-tuberculosis medication’s induced liver injury. Background: Hepatic toxicity is the most serious complication in treatment of tuberculosis. Methods: In a randomized double blind clinical trial (ACTRN12610000643077), 55 cases with hepatotoxicity caused by antituberculosis drugs were divided into two groups. Informed consents were obtained. The intervention group received silymarin and the control group received placebo. Severity of liver injury, the duration necessary for normalization of liver function and hospital stay were compared between the two groups. Results: There was not any statistically significant difference in the rate of adverse effects between silymarin and placebo groups. Conclusion: Although silymarin is considered a safe herbal medication, it was not effective to treat hepatic toxicity of anti-tuberculosis drugs. © 2019 RIGLD.

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Developing rat bone marrow derived mast cells by the splenic cells culture supernatant of rat and mouse

Abstract: Background: Mast cells play a critical role in the pathogenesis of various immunological and non-immunological diseases. It is now accepted that culturing primary mast cells considered as a tool for investigation role of mast cells in diseases. Development of various animal primary mast cells and their function could be used for the translational studies in the pathogenesis of human diseases. The aim of the study was to develop simple and cost-efficient method for differentiation and culture of rat mast cells from bone marrow by using rat and mouse spleen supernatant. Materials and Methods: Bone marrow cells from 10 to15-weeks-old male rats was obtained and cultured for three weeks on cell culture medium. After that, purity of cells was approved by FC?RI and CD117 antibodies, toluidine blue and Immunohistochemistry (IHC). Results: After 3 weeks continuous culturing, high purity of cells was found. CD117, CD34 expression and tryptase were 80.1, 76.89 and 87.9%, respectively by rat splenic supernatant, whereas 85.4, 83.07 and 82.1%, respectively with mouse splenic supernatants. Besides, rat spleen supernatant developed 91.4% and mouse splenocyte supernatant developed 89.7% mast cells based on surface markers. Conclusion: The data presented in this study indicated equal maturation and differentiation of bone marrow derived rat mast cells by using both spleen supernatants. © 2019 NRITLD, National Research Institute of Tuberculosis and Lung Disease, Iran.

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