Bosnian Journal of Basic Medical Sciences <p>The Bosnian Journal of Basic Medical Sciences (BJBMS) reaches readers across a wide range of medical disciplines. The journal publishes basic and translational/clinical research submissions in all biomedical specialties, including Genetics and Molecular biology, Immunology, Microbiology, Pathology, Biochemistry, Pharmacology, Anatomy, Biomaterials, new and emerging research and diagnostic methods, new and emerging medical entities, and others.</p> en-US <p>© Association of Basic Medical Sciences of FBIH.</p> (Faruk Skenderi) (Adnan Sabic) Tue, 20 Aug 2019 00:00:00 +0200 OJS 60 Inflammatory cells in perivascular adipose tissue and the integrity of the tunica media in atherosclerotic coronary arteries <p>Obstructive coronary artery disease (CAD) is characterized by inflammation within the atherosclerotic coronary arteries. Infiltration of inflammatory cells into muscular media can lead to remodeling and weakening of the arterial wall. We examined the relationship between inflammatory infiltration in perivascular adipose tissue (PVAT), state of the external elastic membrane, and the intensity of inflammatory infiltration in the tunica media of coronary arteries obtained by endarterectomy from symptomatic patients with diffuse CAD. We analyzed endarterectomy sequesters from 22 coronary arteries that contained the intima, media, a part of the adventitia, and PVAT in at least one part of the sequester. The coronary arteries were divided into two groups according to the presence or absence of inflammatory infiltration in PVAT. Staining with hematoxylin-eosin and by the Movat's method showed atherosclerotic changes in the intima and media. Immunohistochemistry (anti-leukocyte common antigen [LCA] antibody) was used for the detection of leukocytes. We found a significant positive correlation between inflammatory infiltration in PVAT and preservation of the external elastic membrane of coronary arteries. Furthermore, we found a significant negative correlation between inflammatory infiltration in PVAT and the intensity of inflammatory infiltration in the media. It seems that the integrity of the external elastic membrane and the proinflammatory properties of PVAT restrain inflammatory cells within PVAT. Both effects may prevent the migration of inflammatory cells into the media and delay the development of CAD.</p> Ruda Zorc-Pleskovič, Marjeta Zorc, Dušan Šuput, Aleksandra Milutinović Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Fri, 27 Sep 2019 01:31:08 +0200 The use of cellular matrix in symptomatic knee osteoarthritis <p>Knee osteoarthritis is a degenerative "wear and tear" disorder affecting mainly population over 50 years old. It can also present in younger people, especially after an injury or as a part of other diseases. While many therapeutic options exist for knee osteoarthritis, none of them has the potential to cure this condition. Cellular Matrix represents a combination of natural non-crosslinked hyaluronic acid (HA), thixotropic cell separation gel, and sodium citrate anticoagulant solution. A combination of Cellular Matrix with autologous platelet-rich plasma (A-PRP) is a novel therapeutic approach to the management of knee osteoarthritis. It is assumed that the active components HA and PRP have a synergistic effect contributing to a better therapeutic outcome in patients with knee osteoarthritis. Physiotherapy could provide an additional benefit. This is a retrospective pilot study assessing the potential benefit of Cellular Matrix and A-PRP combined with physiotherapy in the management of chronic knee osteoarthritis. Twenty-five patients were enrolled in the study and injected with three doses of Cellular Matrix combined with A-PRP with a time span of 2 weeks between each injection. All patients received standardized physiotherapy. The results showed that 68% of patients achieved more than 50% improvement in pain, stiffness, and function of the knee joints. There were no adverse reactions. This retrospective pilot study confirmed the positive effect of PRP and HA combination in the management of mild and moderate knee osteoarthritis. These preliminary results need to be verified in randomized control trials.</p> Abir Aly Abbassy, Suad Trebinjac, Nehad Kotb Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Fri, 20 Sep 2019 15:42:21 +0200 A male-specific association between AGTR1 hypermethylation and coronary heart disease <p>The <em>AGTR1</em> gene encodes angiotensin II receptor type 1, which is involved in cardiovascular diseases such as coronary heart disease (CHD). In the current study, we analyzed <em>AGTR1</em> methylation level in a Han Chinese population by SYBR green-based quantitative methylation-specific PCR (qMSP). We collected blood samples from 761 CHD patients and 398 non-CHD controls at the Ningbo First Hospital. A data mining analysis was also performed to explore the association between <em>AGTR1</em> methylation and <em>AGTR1 </em>gene expression, using datasets from the cBioPortal for Cancer Genomics and the Gene Expression Omnibus (GEO) database. Our results showed a significantly higher percentage of methylated reference (PMR) of <em>AGTR1</em> in male CHD patients compared with male non-CHD controls (median PMR: 2.12% vs. 0.59%, <em>p</em> = 0.037). The data mining analysis showed that <em>AGTR1</em> expression was significantly increased in human hepatoma HepG2 cells treated with the demethylation agent 5-aza-2'-deoxycytidine (fold = 3.12, <em>p</em> = 0.009). Further data mining analysis using the cholangiocarcinoma (TCGA, PanCancer Atlas) data indicated an inverse association between <em>AGTR1</em> methylation and <em>AGTR1</em> expression (r = -0.595, <em>p</em> = 1.29E-04). Overall, our results suggest that <em>AGTR1</em> methylation is involved in the regulation of <em>AGTR1</em> gene expression and that <em>AGTR1 </em>hypermethylation is associated with CHD in males. These findings may provide new clues about the pathogenesis of CHD.</p> Xiaojing Li, Nan Wu, Huihui Ji, Yi Huang, Haochang Hu, Jiyi Li, Siyu Mi, Shiwei Duan, Xiaomin Chen Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 19 Sep 2019 10:55:48 +0200 Mesenchymal stem cells from bone marrow regulate invasion and drug resistance of multiple myeloma cells by secreting chemokine CXCL13 <p>Multiple myeloma (MM) is a hematologic cancer arising from plasma cells. Mesenchymal stem cells (MSCs) are a heterogeneous cell population in the bone marrow microenvironment. In this study, we evaluated the regulatory effects of MSCs on the invasion and drug resistance of MM cells U266 and LP-1. Bone marrow samples from MM patients and normal subjects were collected. MSCs were extracted from bone marrow and cultured, and their phenotypes were identified by flow cytometry. The level of CXCL13 in the supernatant of cultured MSCs was detected by ELISA. The protein expression of CXCR5 (a specific receptor of CXCL13) in U266 and LP-1 cells was detected by Western blot. The effects of MSCs on the invasion of U266 and LP-1 cells and the resistance to bortezomib were assessed by Transwell and CCK-8 assay, respectively. The mRNA and protein expressions of BTK, NF-κB, BCL-2, and MDR-1 were detected by RT-PCR and Western blot, respectively. CXCL13 was secreted by MSCs in the bone marrow microenvironment, and the level in MSCs from MM patients was significantly higher than that of healthy subjects. CXCR5 was expressed in both U266 and LP-1 cells. The resistance of MM cells to bortezomib was enhanced by MSCs through CXCL13 secretion. The invasion and proliferation of U266 and LP-1 cells were promoted, and the mRNA and protein expressions of BTK, NF-κB, BCL-2, and MDR-1 were upregulated by MSCs. The basic biological functions of MM cells U266 and LP-1 were affected by MSCs via the CXCL13-mediated signaling pathway. This study provides valuable experimental evidence for clinical MM therapy.</p> Guihua Zhang, Faan Miao, Jinge Xu, Rui Wang Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 17 Sep 2019 10:34:53 +0200 HPV infection screening using surface plasmon resonance in women from Kunming (southwest China) <p>No study examined the frequency of human papillomavirus (HPV) genotypes by surface plasmon resonance (SPR) in southwest China. This was a cross-sectional survey (The Second Affiliated Hospital of Kunming Medical University, 10/2010 to 12/2011) in 150 patients who were hospitalized or volunteered for cervical cancer (CC) screening. A HPV typing kit was used to detect 24 types of HPV by the SPR technique. The HPV-positive rate was 34.8% in women with normal cytology and 92.9% in women with CC. The frequency of HPV16 increased from 9.4% for women with normal cytology to 28.9% for cervical intraepithelial neoplasia (CIN)1, 41.4% for CIN2, 54.1% for CIN3, and 71.4% for CC (<em>p</em> &lt; 0.001). The frequency of HPV18 increased from 0% for women with normal cytology to 2.6% for CIN1, 3.4% for CIN2, 5.4% for CIN3, and 21.4% for CC (<em>p</em> = 0.03). HPV40 was only found in one patient with CC (<em>p</em> = 0.04). There was no relation between HPV genotype and women’s age. In Kunming (southwest China), the frequency of HPV infection was 74.0% among women who underwent CC screening. HPV16 and HPV18 were the two most frequent genotypes. SPR could be of value for the screening of HPV infection.</p> Yang Liu, Yue Quan, Changjun Xu, Yajuan Huang, Daizhu Li, Qing Qing, Chunyi Sun, Honglin Zhou Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Wed, 11 Sep 2019 23:14:00 +0200 The effects of exercise on vascular markers and C-reactive protein among obese children and adolescents: An evidence-based review <p>Numerous studies have evaluated the effects of exercise training on obese children and adolescents. However, the impact of aerobic and/or resistance exercise alone, without any other interventions, on vascular markers and C-reactive protein (CRP) in obese children and adolescents is still not clear. We performed a literature search in Ovid Medline, PubMed, and SCOPUS databases to identify articles on the effects of exercise on vascular markers and CRP among obese children and adolescents, published between January 2009 and May 2019. Only full-text articles in English that reported on the effect of aerobic and/or resistance exercise on the vascular markers pulse wave velocity (PWV), carotid intima-media thickness (CIMT), flow-mediated dilatation (FMD), augmentation index (AIx) or CRP in obese children and adolescents (5–19 years old) were included. The literature search identified 36 relevant articles; 9 articles that fulfilled all the inclusion criteria were selected by two independent reviewers. Aerobic exercise or a combination of aerobic and resistance exercise training significantly improved CIMT and PWV in obese children and adolescents in all studies in which they were measured (2 studies for PWV and 4 studies for CIMT). However, the effects of exercise on FMD and CRP levels were inconclusive, as only half of the studies demonstrated significant improvements (1/2 studies for FMD and 4/8 studies for CRP). The results of our review support the ability of exercise to improve vascular markers such as PWV and CIMT in obese children and adolescents. This finding is important as obesity is a modifiable risk factor of CVD, and exercise may help in reducing the future occurrence of CVD in this population.</p> Norizam Salamt, Musilawati Muhajir, Amilia Aminuddin, Azizah Ugusman Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 10 Sep 2019 15:24:35 +0200 Comparison of the effects of preoperative melatonin or vitamin C administration on postoperative analgesia <p>The analgesic benefit of melatonin and vitamin C as primary or adjuvant agents has been reported in various studies; however, their analgesic effects in the treatment of postoperative pain remain unclear. Thus, we aimed to evaluate the effect of single preoperative dose of oral melatonin or vitamin C administration on postoperative analgesia. In this study, we recruited 165 adult patients undergoing elective major abdominal surgery under general anesthesia. Patients were randomly divided into three equal (n = 55) groups. One hour before surgery, patients received orally melatonin (6 mg) in group M, vitamin C (2 g) in group C, or a placebo tablet in group P. Pain, sedation, patient satisfaction, total morphine consumption from a patient-controlled analgesia device, supplemental analgesic requirement, and the incidence of nausea and vomiting were recorded throughout 24 h after surgery. The mean pain score and total morphine consumption were found significantly lower in both M and C groups compared with group P (<em>p </em>&lt; 0.001). There were no significant differences between group M and C with respect to pain scores (<em>p </em>= 0.117) and total morphine consumption (<em>p </em>= 0.090). Patients requested less supplemental analgesic and experienced less nausea and vomiting in groups M and C compared with group P. In conclusion, preoperative oral administration of 6 mg melatonin or 2 g vitamin C led to a reduction in pain scores, total morphine consumption, supplemental analgesic requirement, and the incidence of nausea and vomiting compared with placebo.</p> Demet Laflı Tunay, Murat Türkeün Ilgınel, Hakkı Ünlügenç, Merthan Tunay, Feride Karacaer, Ebru Biricik Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Fri, 30 Aug 2019 01:02:22 +0200 Tissue-based metabolomics reveals potential biomarkers for cervical carcinoma and HPV infection <p>Aberrant metabolic regulation has been observed in human cancers, but the corresponding regulation in human papillomavirus (HPV) infection-associated cervical cancer is not well understood. Here, we explored potential biomarkers for the early prediction of cervical carcinoma based on the metabolic profile of uterine cervical tissue specimens that were positive for HPV16 infection. Fifty-two fresh cervical tissues were collected from women confirmed to have cervical squamous cell carcinoma (SCC; n = 21) or cervical intraepithelial neoplasia (CIN) stages II-III (n = 20). Eleven healthy women constituted the controls (NCs). Real-time polymerase chain reaction (PCR) was performed to detect HPV infection in the tissues. High-resolution magic angle spinning nuclear magnetic resonance was utilized for the analysis of the metabolic profile in the tissues. The expression of rate-limiting enzymes involved in key metabolic pathways was detected by reverse-transcription quantitative PCR. An independent immunohistochemical analysis was performed using 123 cases of paraffin-embedded cervical specimens. A profile of 17 small molecular metabolites that showed differential expression in HPV16-positive cervical SCC or CIN II-III compared with HPV-negative NCs was identified. According to the profile, the levels of α- and β-glucose decreased, those of lactate and low-density lipoproteins increased, and the expression of multiple amino acids was altered. Significantly increased transcript and protein levels of glycogen synthase kinase 3 beta (GSK3β) and glutamate decarboxylase 1 (GAD1) and decreased transcript and protein levels of pyruvate kinase M2 (PKM2) and carnitine palmitoyltransferase 1A (CPT1A) were observed in the patient group (<em>p </em>&lt; 0.05). HPV infection and cervical carcinogenesis drive metabolic modifications that might be associated with the aberrant regulation of enzymes related to metabolic pathways.</p> Abulizi Abudula, Nuermanguli Rouzi, Lixiu Xu, Yun Yang, Axiangu Hasimu Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 29 Aug 2019 11:18:33 +0200 Pathogenesis of atherosclerosis in the tunica intima, media, and adventitia of coronary arteries: An updated review <p>Atherosclerosis is a chronic inflammatory disease of arteries and it affects the structure and function of all three layers of the coronary artery wall. Current theories suggest that the dysfunction of endothelial cells is one of the initial steps in the development of atherosclerosis. The view that the tunica intima normally consists of a single layer of endothelial cells attached to the subendothelial layer and internal elastic membrane has been questioned in recent years. The structure of intima changes with age and it becomes multilayered due to migration of smooth muscle cells from the media to intima. At this stage, the migration and proliferation of smooth muscle cells do not cause pathological changes in the intima. The multilayering of intima is classically considered to be an important stage in the development of atherosclerosis, but in fact atherosclerotic plaques develop only focally due to the interplay of various processes that involve the resident and invading inflammatory cells. The tunica media consists of multiple layers of smooth muscle cells that produce the extracellular matrix, and this layer normally does not contain microvessels. During the development of atherosclerosis, the microvessels from the tunica adventitia or from the lumen may penetrate thickened media to provide nutrition and oxygenation. According to some theories, the endothelial dysfunction of these nutritive vessels may significantly contribute to the atherosclerosis of coronary arteries. The adventitia contains fibroblasts, progenitor cells, immune cells, microvessels, and adrenergic nerves. The degree of inflammatory cell infiltration into the adventitia, which can lead to the formation of tertiary lymphoid organs, correlates with the severity of atherosclerotic plaques. Coronary arteries are surrounded by perivascular adipose tissue that also participates in the atherosclerotic process.</p> Aleksandra Milutinović, Dušan Šuput, Ruda Zorc-Pleskovič Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 27 Aug 2019 14:35:04 +0200 A preliminary study of microRNA expression in different types of primary melanoma <p>MicroRNAs (miRNAs) have been proven to regulate the development and progression of cancer through various mechanisms. The aim of the present study was to compare miRNA expression between primary melanomas from different sites. We analyzed the expression of 84 miRNAs in 27 primary melanoma and 5 nevus formalin-fixed paraffin-embedded (FFPE) samples using the Human Cancer PathwayFinder miScript miRNA PCR Array. The FFPE samples were obtained from the archives of the Municipal Clinical Emergency Hospital of Timisoara and included 10 cutaneous melanomas, 10 uveal melanomas, 7 mucosal melanomas, and 5 cutaneous nevi. Out of 84 miRNAs, 11 miRNAs showed altered expression in all types of melanoma compared with the nevi. Among these, miR-155-5p, miR-9-5p, miR-142-5p, miR-19a-3p, miR-134-5p, and miR-301a-3p were upregulated, while miR-205-5p, miR-203a-3p, miR-27b-3p, miR-218-5p, and miR-23b-3p were downregulated. The highest similarity in miRNA expression pattern was found between uveal and mucosal melanoma groups, i.e., 15 miRNAs had altered expression in both groups. Overall, we identified several miRNAs with significantly altered expression in primary melanomas, including those reported for the first time in this type of cancer. Among them, mir-9-5p, mir-203a-3p, mir-19a-3p, mir-27b-3p, and mir-218-5p showed altered expression in all three melanoma types vs. nevi. Further research should explore the potential of these miRNAs in melanoma.</p> Ioana Gencia, Flavia Baderca, Stefania Avram, Armand Gogulescu, Anca Marcu, Edward Seclaman, Catalin Marian, Caius Solovan Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 27 Aug 2019 10:39:52 +0200 Effect of SOD2 methylation on mitochondrial DNA4834-bp deletion mutation in marginal cells under oxidative stress <p>Presbycusis, or age-related hearing loss, is a prevalent disease that severely affects the physical and mental health of the elderly. Oxidative stress and mitochondrial (mt)DNA deletion mutation are considered as major factors in the pathophysiology of age-related hearing loss. The 4977-bp deletion in human mtDNA (common deletion, corresponding to the 4834-bp mtDNA deletion in rats) is suggested to be closely associated with the pathogenesis of age-related hearing loss. Superoxide dismutase 2 (SOD2), an isoform of SOD that is exclusively expressed in the intracellular mitochondrial matrix, plays a crucial role in oxidative resistance against mitochondrial superoxide. Previous research has shown that methylation of the promoter region of the <em>SOD2</em> gene decreased the expression of SOD2 in marginal cells (MCs) extracted from the inner ear of rats subjected to D-galactose-induced mtDNA4834 deletion. However, the relationship between <em>SOD2</em> methylation and mtDNA4834 deletion under oxidative stress remains to be elucidated. Herein, an oxidative damage model was established in the extracted MCs using hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), which increased the methylation level of <em>SOD2 </em>and the copy number of mtDNA4834 mutation in MCs. Decreasing the methylation level of <em>SOD2</em> using 5-azacytidine, a DNA methylation inhibitor, reduced oxidative stress and the copy number of mtDNA4834 mutation and inhibited H<sub>2</sub>O<sub>2</sub>-induced apoptosis. The present work demonstrates that decreasing the methylation of <em>SOD2</em> suppresses the mtDNA4834 deletion in MCs under oxidative stress and provides potential insights to the intervention therapy of aging-related hearing loss.</p> Jun Li, Xiang Dai, Xuelian He, Rong Yang, Zhongfang Xia, Han Xiao Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Thu, 22 Aug 2019 13:34:01 +0200 Internodal HER2 heterogeneity of axillary lymph node metastases in breast cancer patients <p>Determination of human epidermal growth factor receptor 2 (HER2) status is important for adequate treatment of breast cancer (BC) patients. The novel HER2 gene protein assay (GPA) is particularly convenient, as it allows the simultaneous assessment of HER2 protein expression and gene amplification at individual cell level. Here we investigated the frequency of internodal HER2 heterogeneity in axillary lymph node macrometastases of BC patients and compared HER2 status between primary breast tumor and its metastases. We included a total of 41 female patients operated between 2014 and 2015 for primary BC with axillary lymph node macrometastases. Representative paraffin blocks of metastatic lymph nodes were sectioned and the slides were stained using the GPA in 38 BC cases. GPA results were assessed according to the ASCO/CAP 2013 criteria. We analyzed 12586 individual tumor cells, 120 cells per section of each metastatic lymph node. HER2 status differed between the primary tumor and its metastases in 5/38 cases (13.2%). In patients with at least two metastatic nodes, the HER2 status of lymph node metastases was only slightly different in 4/23 cases (17.4%). Our results indicate rare but substantial differences in HER2 status between primary breast tumor and its axillary lymph node metastases that may direct the choice and outcomes of targeted therapy in BC patients. The impact of the rare and subtle internodal HER2 heterogeneity evidenced in this study remains uncertain. Determining the HER2 status of lymph node metastases in BC seems to be rational, but assessing a limited number of metastatic nodes may be sufficient.</p> Ilija Vladimir Baroš, Nataša Tanasković, Ulrika Pellas, Živka Eri, Ljiljana Tadić Latinović, Tibor Tot Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 Expression patterns and prognostic value of miR-210, miR-494, and miR-205 in middle-aged and old patients with sepsis-induced acute kidney injury <p>Septic patients suffer a ‘cytokine storm’ from proinflammatory cytokines, chemokines and other inflammatory mediators, resulting in acute kidney injury (AKI) and death. The purpose of the present study was to determine the expression patterns of microRNA-210 (miR-210), miR-494, and miR-205 in middle-aged and old patients with sepsis-induced AKI and to evaluate their association with patient prognosis. Serum blood urea nitrogen (BUN), creatinine (Cr) and cystatin C levels were determined in peripheral venous blood collected from 110 patients with sepsis-induced AKI and 110 healthy controls. The expression profile of 30 miRNAs was analyzed by TaqMan low-density array (TLDA) in plasma samples from patients and controls. Association of miRNAs with prognosis and survival of patients was analyzed by Spearman’s rank correlation coefficient, Cox multivariate analysis, and ROC curve analysis. TILDA analysis showed 11 upregulated and 11 downregulated miRNAs in patients with sepsis-induced AKI. MiR-210 and miR-494 were the most upregulated and miR-205 was the most downregulated miRNA. High expression of miR-210 and miR-494 was positively correlated with BUN, Cr and cystatin C levels of patients, while low expression of miR-205 was negatively correlated. MiR-210 and miR-494 expression was significantly decreased and miR-205 expression was increased in survivors with sepsis-induced AKI (28-day survival, n = 68) vs. non-survivors (n = 42). BUN, Cr, and miR-205 were independent risk factors for prognosis in sepsis-induced AKI. Our study showed the predictive value of miR-210, miR-494, and miR-205 in prognosis and survival of patients with sepsis-induced AKI. MiR-205 is an independent risk factor for sepsis-induced AKI and its decreased expression is associated with shorter patient survival.</p> Yongjun Lin, Ying Ding, Shuping Song, Man Li, Tao Wang, Feng Guo Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 Thymic stromal lymphopoietin levels are increased in patients with celiac disease <p>Thymic stromal lymphopoietin (TSLP) is a cytokine produced by epithelial cells in the lungs, skin, and intestinal mucosa and is involved in several physiological and pathological processes. In this study, we evaluated serum TSLP levels in patients with celiac disease (CD). The prospective study was conducted at a gastroenterology outpatient clinic between March 2018 and August 2018. Eighty-nine participants aged between 18 and 75 years were classified into following groups: 22 patients with newly diagnosed CD; 20 patients with CD who were compliant with a gluten-free diet (GFD); 32 patients with CD who were not compliant with a GFD; and 15 healthy controls. Demographic characteristics, disease duration, and selected biochemical and hematologic parameters were recorded and compared between groups. Median serum TSLP levels were 1193.65 pg/mL (range: 480.1–1547.1) in newly diagnosed CD patients, 110.25 pg/mL (range: 60.3–216.7) in CD patients who were compliant with a GFD, 113.1 pg/mL (range: 76.3–303.4) in CD patients who were not compliant with a GFD, and 57 pg/mL (range: 49–67.8) in healthy controls. Overall, there was a significant difference in serum TSLP levels between groups (<em>p</em> = 0.001). Patients with newly diagnosed CD had the highest serum TSLP levels. There was no significant difference in serum TSLP levels between patients with CD who were and were not compliant with a GFD. TSLP appears to be involved in the pathogenesis of CD. Further studies are required to determine if the TSLP signaling pathway can be used in the treatment of CD.</p> Evrim Kahramanoğlu Aksoy, Muhammet Yener Akpınar, Ferdane Pirinççi Sapmaz, Özlem Doğan, Metin Uzman, Yaşar Nazlıgül Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 Epidemiology and mechanism of drug resistance of Mycoplasma pneumoniae in Beijing, China: A multicenter study <p><em>Mycoplasma pneumoniae</em> (<em>M. pneumoniae</em>) is one of the most common causes of community-acquired respiratory tract infections (RTIs). We aimed to investigate the prevalence of <em>M. pneumoniae </em>infection, antibiotic resistance and genetic diversity of <em>M. pneumoniae</em> isolates across multiple centers in Beijing, China. P1 protein was detected by Nested PCR to analyze the occurrence of <em>M. pneumoniae </em>in pediatric patients with RTI. <em>M. pneumoniae</em> isolates were cultured and analyzed by Nested-PCR to determine their genotypes. Broth microdilution method was used to determine the minimum inhibitory concentration (MIC) of antibiotics. Out of 822 children with RTI admitted to 11 hospitals in Beijing, 341 (41.48%) were positive for <em>M. pneumoniae</em> by Nested PCR and 236 (69.21%) samples had mutations in 23S rRNA domain V. The highest proportion of <em>M. pneumoniae </em>positive samples was observed in school-age children (118/190; 62.11%) and in pediatric patients with pneumonia (220/389; 56.56%). Out of 341 <em>M. pneumoniae</em> positive samples, 99 (12.04%) isolates were successfully cultured and the MIC values were determined for 65 <em>M. pneumoniae</em> strains. Out of these, 57 (87.69%) strains were resistant to macrolides, and all 65 strains were sensitive to tetracyclines or quinolones. <em>M. pneumoniae</em> P1 type I and P1 type II strains were found in 57/65 (87.69%) and 8/65 (12.31%) of cultured isolates, respectively. Overall, we demonstrated a high prevalence of <em>M. pneumoniae </em>infection and high macrolide resistance of<em> M. pneumoniae </em>strains in Beijing. School-age children were more susceptible to<em> M. pneumoniae</em>, particularly the children with pneumonia. Thus, establishment of a systematic surveillance program to fully understand the epidemiology of <em>M. pneumoniae</em> is critical for the standardized use of antibiotics in China.</p> Dong-Xing Guo, Wen-Juan Hu, Ran Wei, Hong Wang, Bao-Ping Xu, Wei Zhou, Shao-Jie Ma, Hui Huang, Xuan-Guang Qin, Yue Jiang, Xiao-Pei Dong, Xiao-Yan Fu, Da-Wei Shi, Liang-Yu Wang, A-Dong Shen, De-Li Xin Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 EP1 receptor is involved in prostaglandin E2-induced osteosarcoma growth <p>Recent studies showed that the activation of prostaglandin (PG) receptor EP1 promotes cell migration and invasion in different cancers. The aim of this study was to investigate the role of EP1 in the proliferation of osteosarcoma (OS) cells <em>in vitro</em> and <em>in vivo</em>. EP1 mRNA and protein levels were analyzed by real-time RT-PCR and Western blot, respectively in human OS cell lines MG63, OS732, U-2OS, and 143B compared to human fetal osteoblastic hFOB 1.19 cells. MG63 cells were treated with PGE<sub>2</sub>, EP1 specific agonist 17-PT-PGE<sub>2</sub>, 17-PT-PGE<sub>2</sub> + EP1 specific antagonist SC51089, or DMSO (control). EP1R-siRNA or a non-silencing irrelevant RNA duplex (negative control) were used for the transfection of MG63 cells, followed by PGE<sub>2</sub> treatment. Nude mice carrying MG63 xenografts were treated with SC51089 (2 mg/kg/day). MG63 cells/xenografts were analyzed by MTT assay, TUNEL assay, PKC enzyme activity assay, and Western blot (EP1 and apoptotic proteins), and tumor growth/volume was evaluated in mice. EP1 levels were significantly higher in OS cells compared to osteoblasts. PGE<sub>2</sub> or 17-PT-PGE<sub>2</sub> treatment increased the proliferation and decreased the apoptosis of MG63 cells. Inhibition of EP1 by SC51089 or siRNA markedly decreased the viability of MG63 cells. Similarly, SC51089 treatment significantly inhibited MG63 cell proliferation and promoted apoptosis <em>in</em> <em>vivo</em>. The silencing of EP1 receptor by siRNA or blockade of EP1 signaling by SC51089 activated extrinsic and intrinsic apoptotic pathways both <em>in</em> <em>vivo</em> and <em>in</em> <em>vitro</em>, as evidenced by increased levels of Bax, cyt c, cleaved caspase-3, caspase-8 and caspase-9. EP1 appears to be involved in PGE<sub>2</sub>-induced proliferative activity of MG63 cells. Antagonizing EP1 may provide a novel therapeutic approach to the treatment of OS.</p> Jing-cai Niu, Nan Ma, Wei Liu, Pei-ji Wang Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 Isoflurane exposure in infant rats acutely increases aquaporin 4 and does not cause neurocognitive impairment <p>Isoflurane is commonly used in pediatric population, but its mechanism of action in cognition is unclear. Aquaporin 4 (AQP4) regulates water content in blood, brain, and cerebrospinal fluid. Various studies have provided evidence for the role of AQP4 in synaptic plasticity and neurocognition. In this study, we aimed to determine whether a prolonged exposure to isoflurane in infant rats is associated with cognition and what effect this exposure has on AQP4 expression. Ten-day-old [postnatal day (P) 10] Wistar albino rats were randomly allocated to isoflurane group (n = 32; 1.5% isoflurane in 50% oxygen for 6 hours) or control group (n = 32; only 50% oxygen for 6 hours). Acute (P11) and long-term (P33) effects of 6-hour anesthetic isoflurane exposure on AQP4 expression were analyzed in whole brains of P11 and P33 rats by RT-qPCR and Western blot. Spatial learning and memory were assessed on P28 to P33 days by Morris Water Maze (MWM) test. The analysis revealed that isoflurane increased acutely both mRNA (~4.5 fold) and protein (~90%) levels of AQP4 in P11 rats compared with control group. The increasing levels of AQP4 in P11 were not observed in P33 rats. Also, no statistically significant change between isoflurane and control groups was observed in the latency to find the platform during MWM training and probe trial. Our results indicate that a single exposure to isoflurane anesthesia does not influence cognition in infant rats. In this case, acutely increased AQP4 after isoflurane anesthesia may have a protective role in neurocognition.</p> Serdar Demirgan, Onat Akyol, Zeynep Temel, Aslıhan Şengelen, Murat Pekmez, Recep Demirgan, Mehmet Salih Sevdi, Kerem Erkalp, Ayşin Selcan Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 PCAT1 is a poor prognostic factor in endometrial carcinoma and associated with cancer cell proliferation, migration and invasion <p>Long non-coding RNAs (lncRNAs) are emerging as important modulators of cancer progression, among which prostate cancer-associated transcript 1 (PCAT1) has been shown to be an oncogene in several tumors. However, the clinical significance and biological function of PCAT1 in endometrial carcinoma (EC) remain unclear. In this study, we used 89 EC tissues and HEC-1B, Ishikawa, RL95-2 and AN3CA EC cell lines. We found elevated expression levels of PCAT1 in EC tissues and cell lines using reverse transcription qPCR (RT-qPCR). The prognostic value of PCAT1 was determined using Kaplan–Meier survival and Cox regression analysis. The results showed that higher PCAT1 expression was positively correlated with FIGO stage, myometrial invasion, lymph node metastasis, and a shorter overall survival. A series of functional assays showed that the knockdown of PCAT1 by small interfering RNA (siRNA) targeting PCAT1 (siPCAT1) suppressed cell proliferation, migration and invasion, but promoted apoptosis. Western blot analysis further showed that B-cell lymphoma 2 (Bcl-2), vimentin and N-cadherin were downregulated, but E-cadherin and Bcl-2-associated death promoter (Bad) were upregulated in PCAT1-silenced EC cells. Taken together, our results underscore the oncogenic role of PCAT1 in EC and show that PCAT1 may be a potential therapeutic target in EC treatment.</p> Xiaohuan Zhao, Yali Fan, Changqiong Lu, Hongfang Li, Ning Zhou, Gaogao Sun, Hong Fan Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 Valspodar-modulated chemotherapy in human ovarian cancer cells SK-OV-3 and MDAH-2774 <p>Overcoming drug resistance in ovarian cancer is the overarching goal in gynecologic oncology. One way to increase drug cytotoxicity without increasing the drug dose is to simultaneously apply multidrug resistance modulator. Valspodar is the second generation P-glycoprotein 1 modulator capable of reversing multidrug resistance in different cancers. In this study, we evaluated the effect of valspodar and cisplatin co-treatment on cell viability, cell death and oxidative status in ovarian cancer cells. Two human ovarian cancer cell lines SK-OV-3 and MDAH-2774 were treated with cisplatin, valspodar, or cisplatin + valspodar for 24 or 48 hours. Untreated cells were used as control group. Cell viability was evaluated by MTT assay. Cell death was assessed by TUNEL and comet assay. Lipid peroxidation (malondialdehyde) and protein thiol groups were analyzed as oxidative stress markers. The expression of mitochondrial superoxide dismutase (MnSOD) was assessed by immunocytochemistry. Valspodar effectively reduced the resistance of SK-OV-3 cells to cisplatin, as demonstrated by increased oxidative stress, decreased cell viability and increased apoptosis in SK-OV-3 cells co-treated with valspodar and cisplatin compared to other groups. However, valspodar did not significantly affect the resistance of MDAH-2774 cells to cisplatin. Stronger staining for MnSOD in MDAH-2774 vs. SK-OV-3 cells after co-treatment with cisplatin and valspodar may determine the resistance of MDAH-2774 cell line to cisplatin.</p> Maciej Zalewski, Julita Kulbacka, Jolanta Saczko, Małgorzata Drag-Zalesinska, Anna Choromanska Copyright (c) 2019 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200 How latent viruses cause breast cancer: An explanation based on the microcompetition model <p>Most breast cancer cases show a decrease in the concentration of the breast cancer type 1 susceptibility protein (BRCA1). However, only a small portion of these cases have a mutated <em>BRCA1</em> gene. Although many attempts have been made to identify the reason for the decrease in BRCA1 concentration in sporadic, non-heritable breast cancer cases, the cause is still unknown. In this review, we use the Microcompetition Model to explain how certain latent viruses, which are frequently detected in breast cancer tumors, can decrease the expression of the <em>BRCA1</em> gene and cause the development of breast tumors.</p> Hanan Polansky, Hava Schwab Copyright (c) 2018 Bosnian Journal of Basic Medical Sciences Tue, 20 Aug 2019 00:00:00 +0200