Bosnian Journal of Basic Medical Sciences 2019-03-22T15:01:12+01:00 Faruk Skenderi Open Journal Systems <p>The Bosnian Journal of Basic Medical Sciences (BJBMS) is an international, English-language, peer-reviewed journal, publishing articles in different disciplines of basic medical sciences. BJBMS welcomes original research and comprehensive reviews as well as short research communications in the field of molecular biology, biochemistry, genetics, immunology, microbiology, pathology, pharmacology, pharmaceutical sciences, physiology and translational research.</p> Histological and biochemical effects of Cinnamomum cassia nanoparticles in kidneys of diabetic Sprague-Dawley rats 2019-03-22T15:01:11+01:00 Koffi Kouame Aniekan Imo Peter Edidiong Nnamso Akang Roshila Moodley Edwin Coleridge Naidu Onyemaechi Okpara Azu <p>This study investigated the antidiabetic activity of<em> Cinnamomum cassia</em> (<em>C. cassia</em>, Cc) silver nanoparticles (CcAgNPS) and effects of <em>C. cassia</em> on the kidneys of rats with induced type 2 diabetes. Twenty-four Sprague-Dawley rats weighing 250 ± 20 g were induced with diabetes by intraperitoneal injection of streptozotocin (STZ, 60 mg/kg). Animals were randomly assigned to one of four groups (n = 6) and treated for eight weeks with normal saline (control, group A), 5 mg/kg of CcAgNPs (group B), 10 mg/kg of CcAgNPs (group C), or 200 mg/kg of Cc (group D). Body weight and fasting blood glucose (FBG) was measured weekly and fortnightly, respectively. At the end of experiments animals were euthanized, blood and kidney tissue samples were collected for biochemistry (oxidative stress markers and renal function parameters) and kidneys were harvested for histology (PAS and H&amp;E staining). Body weight was significantly higher in group B and C vs. control (<em>p</em> &lt; 0.05), while no significant differences were observed in the kidney-to-body weight ratio between groups. FBG, glutathione, malondialdehyde, alanine aminotransferase, aspartate aminotransferase, serum urea and creatinine were significantly lower in group B, C and/or D vs. control (all <em>p</em> &lt; 0.05). In group A, severe distortion of the glomerular network was observed, marked by the loss of capsular integrity, thickened basement membrane, tubular cells with pyknotic nuclei, vacuolization, and interstitial space with infiltrations. These adverse effects were mitigated by 5 mg/kg and 10 mg/kg of CcAgNPs. Our study confirms structural and functional damage to kidneys caused by diabetes. CcAgNPs have a regenerative potential in diabetes-induced kidney damage and may be used as an antidiabetic agent.</p> 2019-03-22T10:55:49+01:00 ##submission.copyrightStatement## Recombinant deoxyribonucleoside kinase from Drosophila melanogaster can improve gemcitabine based combined gene/chemotherapy for targeting cancer cells 2019-03-22T15:01:12+01:00 Mahak Fatima M. Mubasshar Iqbal Ahmed Faiza Batool Anjum Riaz Moazzam Ali Birgitte Munch-Petersen Zeeshan Mutahir <p>A recombinant deoxyribonucleoside kinase from <em>Drosophila melanogaster</em> with a deletion of the last 20 amino acid residues (named <em>Dm</em>dNKΔC20) was hypothesized as a potential therapeutic tool for gene therapy due to its broad substrate specificity and better catalytic efficiency towards nucleosides and nucleoside analogs. This study was designed to evaluate the effect of <em>Dm</em>dNKΔC20 for sensitizing human cancer cell lines towards gemcitabine and to further investigate its role in reversal of acquired drug resistance in gemcitabine-resistant cancer cell line. The <em>DmdNKΔC20</em> gene was delivered to three different cancer cell lines, including breast, colon and liver cancer cells, using lipid-mediated transfection reagent. After transfection, gene expression of <em>DmdNKΔC20 </em>was confirmed by reverse transcription quantitative PCR (qRT-PCR) and the combined effect of <em>Dm</em>dNKΔC20 and gemcitabine based cytotoxicity was observed by cell viability assay. We further evolved a gemcitabine-resistant breast cancer cell line (named MCF7-R) through directed evolution in the laboratory, which showed 375-fold more resistance compared to parental MCF7 cells. Upon transfection with <em>DmdNKΔC20</em> gene, MCF7-R cells showed 83-fold higher sensitivity to gemcitabine compared to the control group of MCF7-R cells. Moreover, we observed 79% higher expression of p21 protein in transfected MCF7-R cells, which may indicate induction of apoptosis. Our findings highlight the importance and therapeutic potential of <em>DmdNKΔC20 </em>in combined gene/chemotherapy approach to target a wide range of cancers<em>,</em> particularly gemcitabine-resistant cancers.</p> 2019-03-19T09:18:36+01:00 ##submission.copyrightStatement## Epidemiology and mechanism of drug resistance of Mycoplasma pneumoniae in Beijing, China: A multi-center study 2019-03-21T15:01:04+01:00 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 <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> 2019-03-17T21:06:48+01:00 ##submission.copyrightStatement## Soft tissue grafts for dural reconstruction after meningioma surgery 2019-03-21T15:01:04+01:00 Tomaz Velnar Lidija Gradisnik <p>The meninges are involved in various pathologies and are often directly or indirectly severed during surgical procedures, especially the dura mater. This can pose a real challenge for the surgeon, as a proper reconstruction of the meninges is important to prevent complications such as cerebrospinal fluid leak (CSF). A variety of techniques for dural reconstruction have been described, employing natural and artificial materials. A novel technique for dural reconstruction involves soft tissue grafts in the form of fibrous or fibromuscular flaps, which are placed on the dural defects to seal the gaps. These soft tissue grafts represent an appropriate scaffold for cell ingrowth and fibrosis, thus preventing CSF. In this pilot study, we described the application of soft tissue grafts for dural reconstruction in 10 patients who underwent convexity meningioma surgery.</p> 2019-03-11T11:50:29+01:00 ##submission.copyrightStatement## Outcomes and prognostic factors for patients with cervical esophageal cancer undergoing definitive radiotherapy or chemoradiotherapy 2019-03-21T15:01:05+01:00 Xin-xin Du Rong Yu Zhen-fei Wang De-cheng Du Qiao-yun Liu Run-mei Wang Shi-rong Kang Hao Yang <p>Cervical esophageal cancer (CEC) is uncommon, accounting for less than 5% of all esophageal cancers. The management of CEC is controversial. This study investigated treatment outcomes and prognostic factors of survival in CEC patients undergoing definitive radiotherapy or concurrent chemoradiotherapy (CCRT). Ninety-one CEC patients were treated by intensity-modulated radiation therapy (IMRT) and three-dimensional conformal radiation therapy (3DCRT) between July 2007 and September 2017. The mean prescription dose was 64 Gy (range 54-70 Gy) delivered as 1.8-2.2 Gy per fraction per day, 5 days a week. Out of 91 patients, 34 received concurrent cisplatin-based chemotherapy (CT) including 18 patients who also received neoadjuvant CT. Overall survival (OS), locoregional failure-free survival (LRFFS), and progression-free survival (PFS) were estimated by the Kaplan–Meier method. Prognostic factors of survival were determined in univariate (log-rank test) and multivariate (Cox proportional hazard model) analysis. Treatment-related toxicity was also assessed. Median follow-up time for all patients was 19 months. Two-year OS, LRFFS and PFS of all patients were 58.2%, 52.5% and 48.1%, respectively. Clinical stage was an independent prognostic factor for OS (HR = 2.35, 95% CI: 1.03-5.37, <em>p</em> = 0.042), LRFFS (HR = 3.84, 95% CI: 1.38-10.69, <em>p</em> = 0.011), and PFS (HR = 2.68, 95% CI: 1.11-6.45, <em>p</em> = 0.028). Hoarseness was an independent prognostic factor for OS (HR = 2.10, 95% CI: 1.05-4.19, <em>p</em> = 0.036). CCRT was independently associated with better LRFFS (HR = 0.33, 95% CI: 0.14-0.79, <em>p</em> = 0.012). 3DCRT and IMRT with concurrent CT is well-tolerated and may improve local tumor control in CEC patients. Advanced clinical stage and hoarseness are adverse prognostic factors for OS, LRFFS, and PFS in CEC.</p> 2019-03-11T10:04:00+01:00 ##submission.copyrightStatement## Thymic stromal lymphopoietin levels are increased in patients with celiac disease 2019-03-21T15:01:05+01:00 Evrim Kahramanoğlu Aksoy Muhammet Yener Akpınar Ferdane Pirinççi Sapmaz Özlem Doğan Metin Uzman Yaşar Nazlıgül <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 or 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> 2019-03-01T11:43:19+01:00 ##submission.copyrightStatement## Upregulated expression of MNX1-AS1 long noncoding RNA predicts poor prognosis in gastric cancer 2019-03-21T15:01:05+01:00 Wei Zhang Lunhua Huang Xinyang Lu Kecheng Wang Xiaofei Ning Zhiqiang Liu <p>As important regulators of gene expression long noncoding RNAs (lncRNAs) are implicated in various physiological and pathological processes, including cancer. An oncogenic role of MNX1 antisense RNA 1 (MNX1-AS1) lncRNA has been suggested in cervical cancer and glioblastoma. In this study, we investigated the clinicopathological significance and biological function of MNX1-AS1 in gastric cancer (GC). The expression of MNX1-AS1 was analyzed by qRT-PCR in 96 GC and adjacent non-tumor tissues in relation to clinicopathological features and overall survival (OS) of patients, and in five human GC cell lines compared to a normal gastric epithelial cell line. Loss-of-function experiments using small interfering RNA (siRNA) targeting MNX1-AS1 (si-MNX1-AS1) were carried out in AGS and MGC-803 GC cell lines. Cell proliferation (CCK-8 assay), migration (Transwell) and invasion (Transwell Matrigel), and protein expression of proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, vimentin and matrix metallopeptidase 9 (MMP-9) were analyzed in transfected GC cells. Expression of MNX1-AS1 was significantly higher in GC vs. adjacent non-tumor tissues. Higher MNX1-AS1 expression was significantly associated with tumor size, TNM stage and lymph node metastasis. Kaplan–Meier analysis showed that GC patients with higher MNX1-AS1 expression had worse OS compared to patients with lower MNX1-AS1 expression. Multivariate analysis showed that MNX1-AS1 is an independent poor prognostic factor in GC. Knockdown of MNX1-AS1 significantly inhibited proliferation, migration and invasion of AGS and MGC-803 cells, and resulted in increased E-cadherin and decreased PCNA, N-cadherin, vimentin and MMP-9 expression. Taken together, these results suggest that MNX1-AS1 has an oncogenic function in GC and potential as a molecular target in GC therapy.</p> 2019-02-26T02:17:58+01:00 ##submission.copyrightStatement## Isoflurane exposure in infant rats acutely increases aquaporin 4 and does not cause neurocognitive impairment 2019-03-21T15:01:07+01:00 Serdar Demirgan Onat Akyol Zeynep Temel Aslıhan Şengelen Murat Pekmez Recep Demirgan Mehmet Salih Sevdi Kerem Erkalp Ayşin Selcan <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 to control group. The increasing levels of AQP4 in P11 were not observed in P33. 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> 2019-02-26T01:37:41+01:00 ##submission.copyrightStatement## Enhancement of bone consolidation using high-frequency pulsed electromagnetic fields (HF-PEMFs): An experimental study on rats 2019-03-21T15:01:07+01:00 Daniel Oltean-Dan Gabriela Bombonica Dogaru Dragos Apostu Alexandru Mester Horea Rares Ciprian Benea Mihai Gheorghe Paiusan Catalin Ovidiu Popa Elena Mihaela Jianu Gyorgy Istvan Bodizs Cristian Berce Alina Mihaela Toader Gheorghe Tomoaia <p><em>In vitro</em> studies showed that high-frequency pulsed electromagnetic fields (HF-PEMFs) increase the activity/expression of early and late osteogenic markers and enhance bone mineralization. The main aim of this study was to investigate the <em>in vivo</em> effects of HF-PEMFs on fracture healing using a rat model. A femur fracture was established by surgery in 20 male Wistar rats. Titanium nails were implanted to reduce and stabilize the fracture. After surgery, 20 rats were equally divided into untreated control and treated group (from the first postoperative day HF-PEMFs at 400 pulses/sec [pps] were applied for 10 minutes/day, for two weeks). Quantitative and qualitative assessment of bone formation was made at two and eight weeks following surgery and included morphological and histological analysis, serological analysis by ELISA, micro-computed tomography (micro-CT), and three-point bending test. At two weeks in HF-PEMF group, soft callus was at a more advanced fibrocartilaginous stage and the bone volume/total tissue volume (BV/TV) ratio in the callus area was significantly higher compared to control group (<em>p</em> = 0.047). Serum concentration of alkaline phosphatase (ALP) and osteocalcin (OC) was significantly higher in HF-PEMF group (ALP <em>p</em> = 0.026, OC <em>p</em> = 0.006) as well as the mechanical strength of femurs (<em>p</em> = 0.03). At eight weeks, femurs from HF-PEMF group had a completely formed woven bone with dense trabeculae, active bone marrow, and had a significantly higher BV/TV ratio compared to control (<em>p</em> = 0.01). HF-PEMFs applied from the first postoperative day, 10 minutes/day for two weeks, enhance bone consolidation in rats, especially in the early phase of fracture healing.</p> 2019-02-20T13:51:44+01:00 ##submission.copyrightStatement## Vascular endothelial growth factor (VEGF)-related polymorphisms rs10738760 and rs6921438 are not risk factors for proliferative diabetic retinopathy (PDR) in patients with type 2 diabetes mellitus (T2DM) 2019-03-21T15:01:11+01:00 Jana Sajovic Ines Cilenšek Sara Mankoč Špela Tajnšek Tanja Kunej Daniel Petrovič Mojca Globočnik Petrovič <p>Vascular endothelial growth factor (VEGF) is an important regulator of angiogenesis and has been investigated as a candidate gene in a number of conditions, including diabetes and its microvascular complications (e.g., retinopathy and nephropathy). Several <em>VEGF</em>-related polymorphisms have been shown to contribute to nearly half of the variability in circulating VEGF levels in healthy individuals. Our aim was to assess the association between <em>VEGF</em>-related rs10738760 and rs6921438 polymorphisms and proliferative diabetic retinopathy (PDR) in Slovenian patients with type 2 diabetes mellitus (T2DM). We also investigated the effect of these polymorphisms on VEGF receptor 2 (VEGFR-2) expression in fibrovascular membranes (FVMs) from patients with PDR. This case-control study enrolled 505 unrelated patients with T2DM: 143 diabetic patients with PDR as a study group, and 362 patients with T2DM of &gt;10 years duration and with no clinical signs of PDR as a control group. Patient clinical and laboratory data were obtained from their medical records. rs10738760 and rs6921438 polymorphisms were genotyped using TaqMan SNP Genotyping assay. VEGFR-2 expression was assessed by immunohistochemistry in 20 FVMs from patients with PDR, and numerical areal density of VEGFR-2-positive cells was calculated. The occurrence of PDR was 1.7 times higher in diabetic patients carrying GA genotype of rs6921438 compared to patients with GG genotype, with a borderline statistical significance (OR = 1.7, 95% CI = 1.00 – 2.86, <em>p</em> = 0.05). In addition, A allele of rs6921438 was associated with increased VEGFR-2 expression in FVMs from PDR patients. However, we observed no association between AA genotype of rs6921438 nor between rs10738760 variants and PDR, indicating that the two polymorphisms are not genetic risk factors for PDR.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## The association of early postoperative lactate levels with morbidity after elective major abdominal surgery 2019-03-21T15:01:12+01:00 Jelena Veličković Ivan Palibrk Biljana Miličić Dejan Veličković Bojan Jovanović Goran Rakić Milorad Petrović Vesna Bumbaširević <p>Lactate levels are widely used as an indicator of outcome in critically ill patients. We investigated the prognostic value of postoperative lactate levels for postoperative complications (POCs), mortality and length of hospital stay after elective major abdominal surgery. A total of 195 patients were prospectively evaluated. Lactate levels were assessed on admission to the intensive care unit (ICU) [L<sub>0</sub>], at 4 hours (L<sub>4</sub>), 12 hours (L<sub>12</sub>), and 24 hours (L<sub>24</sub>) after the operation. Demographic and perioperative clinical data were collected. Patients were monitored for complications until discharge or death. Receiver operating characteristic (ROC) curves were used to determine the predictive value of lactate levels for postoperative outcomes. The best cut-off lactate values were calculated to differentiate between patients with and without complications, and outcomes in patients with lactate levels above and below the cut-off thresholds were compared. Univariate and multivariate analyses were used to identify variables associated with POCs and mortality. Seventy-six patients developed 184 complications (18 deaths), while 119 had no complications. Serum lactate levels were higher in patients with complications at all time points compared to those without complications (<em>p</em> &lt; 0.001). L<sub>12</sub> had the highest predictive value for complications (AUROC<sub>12</sub> = 0.787; 95% CI: 0.719–0.854; <em>p</em> &lt; 0.001) and mortality (AUROC<sub>12</sub> = 0.872; 95% CI: 0.794–0.950; <em>p</em> &lt; 0.001). The best L<sub>12 </sub>cut-off value for complications and mortality was 1.35 mmol/l and 1.85 mmol/l, respectively. Multivariate analysis revealed that L<sub>12</sub> ≥ 1.35 mmol/l was an independent predictor of postoperative morbidity (OR 2.58; 95% CI 1.27–5.24, <em>p</em> = 0.001). L<sub>24 </sub>was predictive of POCs after major abdominal surgery. L<sub>12</sub> had the best power to discriminate between patients with and without POCs and was associated with a longer hospital stay.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Delirium following total joint replacement surgery 2019-03-21T15:01:10+01:00 Jeffrey Huang Juraj Sprung Toby N. Weingarten <p>Postoperative delirium (POD) is a common complication associated with increased resource utilization, morbidity and mortality. Our institution screens all postsurgical patients for postoperative delirium. The study aim was to perform an automated interrogation of the electronic health records to estimate the incidence of and identify associated risk factors for POD following total joint arthroplasty (TJA). Adult patients who underwent TJA with a multimodal analgesia protocol, including peripheral nerve blockade, from 2008 through 2012, underwent automated chart review. POD was identified by routine nursing assessment and administrative billing codes. Of 11,970 patients, 181 (1.5%) were identified to have POD. Older age (odds ratio, 95% CI 2.20, 1.80–2.71 per decade, <em>p</em> &lt; 0.001), dementia (7.44, 3.54–14.60, <em>p</em> &lt; 0.001), diabetes mellitus (1.70, 1.1.5–2.47, <em>p</em> = 0.009), renal disease (1.68, 1.03–2.65, <em>p</em> = 0.039), blood transfusions (2.04, 1.14–3.52, <em>p</em> = 0.017), and sedation during anesthesia recovery (1.76, 1.23–2.51, <em>p</em> = 0.002) were associated with POD. Anesthetic management was not associated with POD risk. Patients who developed POD required greater healthcare resources. Dementia is strongly associated with POD. The association between POD and transfusions may reflect higher acuity patients or detrimental effect of blood. Postoperative sedation should be recognized as a warning sign of increased risk.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Decreased heart rate recovery may predict a high SYNTAX score in patients with stable coronary artery disease 2019-03-21T15:01:09+01:00 Sadık Volkan Emren Rahman Bilal Gediz Oktay Şenöz Uğur Karagöz Ersin Çağrı Şimşek Fatih Levent Emre Özdemir Mustafa Ozan Gürsoy Cem Nazlı <p>An impaired heart rate recovery (HRR) has been associated with increased risk of cardiovascular events, cardiovascular, and all‐cause mortality. However, the diagnostic ability of HRR for the presence and severity of coronary artery disease (CAD) has not been clearly elucidated. Our aim was to investigate the relationship between HRR and the SYNTAX (SYNergy between percutaneous coronary intervention with TAXus and cardiac surgery) score in patients with stable CAD (SCAD). A total of 406 patients with an abnormal treadmill exercise test and ≥50% coronary stenosis on coronary angiography were included. The HRR was calculated by subtracting the HR in the first minute of the recovery period from the maximum HR during exercise. The SYNTAX score ≥23 was accepted as high. Correlation of HRR with SYNTAX score and independent predictors of high SYNTAX score were determined. A high SYNTAX score was present in 172 (42%) patients. Mean HRR was lower in patients with a high SYNTAX score (9.8 ± 4.5 vs. 21.3 ± 9, <em>p</em> &lt; 0.001). The SYNTAX score was negatively correlated with HRR (r: -0.580, <em>p</em> &lt; 0.001). In multivariate logistic regression analysis, peripheral arterial disease (OR: 13.3; 95% CI: 3.120–34.520; <em>p</em> &lt; 0.001), decreased HRR (OR: 0.780; 95% CI: 0.674–0.902; <em>p</em> = 0.001), peak systolic blood pressure (OR: 1.054; 95% CI: 1.023–1.087; <em>p</em> = 0.001), and peak HR (OR: 0.950; 95% CI: 0.923–0.977; <em>p</em> &lt; 0.001) were found to be independent predictors of a high SYNTAX score. Our results showed that HRR is significantly correlated with the SYNTAX score, and a decreased HRR is an independent predictor of a high SYNTAX score in patients with SCAD.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Predictors of acute kidney injury (AKI) in high-risk ST-elevation myocardial infarction (STEMI) patients: A single-center retrospective observational study 2019-03-21T15:01:08+01:00 Andreja Sinkovič Klara Masnik Matic Mihevc <p>Acute kidney injury (AKI) is a frequent complication in ST-elevation myocardial infarction (STEMI) patients. Factors other than contrast exposure have been suggested as major contributors to renal dysfunction in patients undergoing primary percutaneous coronary intervention (PPCI). Our aim was to assess the incidence and risk factors of AKI in high-risk STEMI patients, mostly treated by PPCI with implemented measures to prevent contrast-induced AKI. We retrospectively analyzed data of 245 STEMI patients (165 men, mean age 63.9 ± 11.9 years) admitted to the Department of Medical Intensive Care Unit. Demographic, clinical, and mortality data were compared between AKI and non-AKI group. AKI was defined as a 1.5-fold increase in serum creatinine from baseline level within 24–48 hours. AKI developed in 34/245 (13.9%) patients. PPCI was performed in 226/245 (92.2%) of all STEMI cases, with no difference between AKI and non-AKI group. There were significant differences between AKI and non-AKI group in diabetes mellitus (41.2% vs. 20.9%), prior MI (26.5% vs. 11.8%), prior resuscitation (38.2% vs. 12.4%), admission acute heart failure [AHF] (44.1% vs. 12.8%), in-hospital AHF (70.6% vs. 17.5%), and hospital-acquired infection [HAI] (79.4% vs. 18.0%). Significantly more AKI patients had increased admission CRP ≥25 mg/L (38.2% vs. 11.8%), peak CRP ≥50 mg/L (91.2% vs. 36%), admission troponin I ≥10 mg/L (44.1% vs. 24.6%), peak troponin I ≥50 mg/L (64.7% vs. 44.1%), peak NT-proBNP ≥400 pmol/L (82.4% vs. 27.5%), and ejection fraction &lt;45% (76.5% vs. 33.6%). Mortality was significantly increased in AKI group, including in-hospital (52.9% vs. 7.1%), 30-day (64.7% vs. 10.7%) and 6-month mortality (70.6% vs. 13.7%). Significant independent predictors of AKI were prior resuscitation (OR 4.171, 95% CI 1.088–15.998), HAI (OR 7.974, 95% CI 1.992–31.912), and peak NT-proBNP (OR 21.261, 95% CI 2.357–191.795). To reduce the risk of AKI in STEMI patients, early diagnosis and treatment of AHF and HAIs are advisable.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Analysis of multidrug-resistant bacteria in 3223 patients with hospital-acquired infections (HAI) from a tertiary general hospital in China 2019-03-21T15:01:08+01:00 Meng Wang Hongyan Wei Yaxin Zhao Linlin Shang Linlin Di Chuanfeng Lyu Jun Liu <p align="left">The frequency of antimicrobial resistance has increased globally due to misuse and overuse of antibiotics, and multi-drug resistant (MDR) bacteria are now recognized as a major cause of hospital-acquired infections (HAI). Our aim was to investigate the prevalence, distribution, and antimicrobial susceptibility rates of MDR bacteria in patients with HAI from a tertiary hospital in China. We retrospectively evaluated all patients with a confirmed diagnosis of bacterial infection at a tertiary general hospital in Jining, for the period between January 2012 and December 2014. The following clinical and demographic data were collected: age, sex, specimens, treatment, microbiology results, and antibiotic resistance patterns of isolates. Bacterial identification and susceptibility testing were performed using VITEK 2 COMPACT system. We screened a total of 15,588 patients, out of which 7579 (48.6%) had an HAI. MDR showed 3223 out of 7579 isolates (42.5%). The most frequently isolated MDR bacteria in patients with HAI were extended-spectrum beta-lactamase (ESBL)-producing <em>Escherichia coli</em> (n = 1216/3223, 37.7%), MDR <em>Pseudomonas aeruginosa </em>(n = 627/3223, 19.5%) and MDR <em>Acinetobacter baumannii </em>(n = 588/3223, 18.2%). MDR-HAI were more common in males (2074/3223, 64.4%) and in elderly patients (≥60 years; 1196/3223, 37.1%). Sputum was the main source of MDR isolates (2056/3223, 63.8%). Patients with MDR-HAI were predominantly distributed in different types of intensive care units. MDR strains in our study showed resistance to most current antibiotics. Overall, patients with HAI infections attributed to MDR bacteria were widely distributed in our hospital. Enhanced surveillance of MDR bacteria is critical for guiding the rational use of antibiotics and reducing the incidence of HAI.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Overexpression of miR-361-5p in triple-negative breast cancer (TNBC) inhibits migration and invasion by targeting RQCD1 and inhibiting the EGFR/PI3K/Akt pathway 2019-03-21T15:01:11+01:00 Jianjun Han Jingjing Yu Yu'na Dai Jumei Li Meiyan Guo Jingzhen Song Xuefeng Zhou <p align="left">Triple-negative breast cancer (TNBC) is the leading cause of cancer-related death in women. Previous studies indicated that miR-361-5p was downregulated in breast cancer, however, the exact effect of miR-361-5p on TNBC requires further investigation. In the present study, we investigated whether miR-361-5p can act as a tumor suppressor by targeting required for cell differentiation 1 homolog (RQCD1) and inhibiting epidermal growth factor receptor (EGFR)/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway in TNBC. The expression of miR-361-5p and RQCD1 was determined by quantitative reverse transcription PCR (qRT-PCR) and/or western blot in TNBC and the adjacent tissues. miR-361-5p mimics were constructed and transfected to TNBC cell line MDA-MB-231. Cells were divided into three groups: blank control group, miRNA mimic negative control (NC) group, and miR-361-5p mimics group. Expression of miR-361-5p, mRNA and protein expression of PI3K, Akt, EGFR, phosphorylated (p)-EGFR/PI3K/Akt, and protein expression of RQCD1 and matrix metallopeptidase 9 (MMP-9) in MDA-MB-231 were measured by qRT-PCR/western blot after transfection. Cell viability was determined by CCK-8 assay. Cell migration and invasion ability were evaluated by scratch and transwell assay, respectively. miR-361-5p target gene was determined by bioinformatics analysis and luciferase reporter assay. RQCD1 was identified as a target of miR-361-5p by TargetScan and confirmed by luciferase reporter assay. Downregulated miR-361-5p and upregulated RQCD1 were observed in TNBC tissues. Expression of EGFR, PI3K, Akt and MMP-9 was inhibited in cells treated with miR-361-5p mimics. Transfection of miR-361-5p mimics also inhibited the phosphorylation of EGFR, PI3K, and Akt. Suppressed cell viability, migration, and invasion was found in miR-361-5p mimics groups. Our results indicated that overexpression of miR-361-5p might act as a suppressor in TNBC by targeting RQCD1 to inhibit the EGFR/PI3K/Akt signaling pathway.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Human amniotic fluid stem cells (hAFSCs) expressing p21 and cyclin D1 genes retain excellent viability after freezing with (dimethyl sulfoxide) DMSO 2019-03-21T15:01:13+01:00 Shiva Gholizadeh-Ghaleh Aziz Zahra Fardyazar Fatima Pashaei-Asl Mohammad Rahmati-Yamchi Khodadad Khodadadi Maryam Pashaiasl <p>Human amniotic fluid stem cells (hAFSCs) have features intermediate between embryonic and adult SCs, can differentiate into lineages of all three germ layers, and do not develop into tumors <em>in vivo</em>. Moreover, hAFSCs can be easily obtained in routine procedures and there is no ethical or legal limitations regarding their use for clinical and experimental applications. The aim of this study was to assess the effect of slow freezing/thawing and two different concentrations of DMSO (10% DMSO + 90% fetal bovine serum [FBS] and 5% DMSO + 95% FBS) on the survival of hAFSCs. hAFSCs were obtained from 5 pregnant women during amniocentesis at 16–22 weeks of gestation. The expression of pluripotency markers (Octamer-binding transcription factor 4 [Oct4] and NANOG) by reverse transcription polymerase chain reaction and cell surface markers (cluster of differentiation [CD31], CD44, CD45, and CD90) by flow cytometry was analyzed before and after the slow-freezing. Cell viability was assessed by trypan blue exclusion or MTT assay. Quantitative mRNA expression of Oct4, NANOG, cyclin D1 and p21 was determined by real-time PCR before and after the slow-freezing. Pluripotency of hAFSCs was confirmed by <em>NANOG</em> and <em>POU5F1</em> (Oct4) gene expression before and after slow-freezing. All hAFSC cultures were positive for CD44 and CD90. A higher viability of hAFSCs was observed after freezing with 90% FBS + 10% DMSO. There was increased expression of <em>NANOG</em> and decreased expression of <em>POU5F1 </em>gene after freezing, compared to control cells (before freezing). DMSO and the process of freezing did not significantly change the expression of p21 and cyclin D1 genes in hAFSCs. Overall, our results indicate the applicability of slow-freezing and DMSO in cryopreservation of SCs.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## MicroRNAs: Recent insights towards their role in male infertility and reproductive cancers 2019-03-21T15:01:11+01:00 Muhammad Babar Khawar Rabia Mehmood Nabila Roohi <p>Spermatogenesis is a tightly controlled, multi-step process in which mature spermatozoa are produced. Disruption of regulatory mechanisms in spermatogenesis can lead to male infertility, various diseases of male reproductive system, or even cancer. The spermatogenic impairment in infertile men can be associated with different etiologies, and the exact molecular mechanisms are yet to be determined. MicroRNAs (miRNAs) are a type of non-protein coding RNAs, about 22 nucleotides long, with an essential role in post-transcriptional regulation. miRNAs have been recognized as important regulators of various biological processes, including spermatogenesis. The aim of this review is to summarize the recent literature on the role of miRNAs in spermatogenesis, male infertility and reproductive cancers, and to evaluate their potential in diagnosis, prognosis and therapy of disease. Experimental evidence shows that aberrant expression of miRNAs affects spermatogenesis at multiple stages and in different cell types, most often resulting in infertility. In more severe cases, dysregulation of miRNAs leads to cancer. miRNAs have enormous potential to be used as diagnostic and prognostic markers as well as therapeutic targets in male infertility and reproductive system diseases. However, to exploit this potential fully, we need a better understanding of miRNA-mediated regulation of spermatogenesis, including the characterization of yet unidentified miRNAs and related regulatory mechanisms.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## A plea for an extension of the anatomical nomenclature: Organ systems 2019-03-21T15:01:12+01:00 Vladimir Musil Alzbeta Blankova Vlasta Dvorakova Radovan Turyna Vaclav Baca <p>This article is the third part of a series aimed at correcting and extending the anatomical nomenclature. Communication in clinical medicine as well as in medical education is extensively composed of anatomical, histological, and embryological terms. Thus, to avoid any confusion, it is essential to have a concise, exact, perfect and correct anatomical nomenclature. The Terminologia Anatomica (TA) was published 20 years ago and during this period several revisions have been made. Nevertheless, some important anatomical structures are still not included in the nomenclature. Here we list a collection of 156 defined and explained technical terms related to the anatomical structures of the human body focusing on the digestive, respiratory, urinary and genital systems. These terms are set for discussion to be added into the new version of the TA.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement## Cannabinoids in cancer treatment: Therapeutic potential and legislation 2019-03-21T15:01:10+01:00 Barbara Dariš Mojca Tancer Verboten Željko Knez Polonca Ferk <p align="left">The plant <em>Cannabis sativa</em> L. has been used as an herbal remedy for centuries and is the most important source of phytocannabinoids. The endocannabinoid system (ECS) consists of receptors, endogenous ligands (endocannabinoids) and metabolizing enzymes, and plays an important role in different physiological and pathological processes. Phytocannabinoids and synthetic cannabinoids can interact with the components of ECS or other cellular pathways and thus affect the development/progression of diseases, including cancer. In cancer patients, cannabinoids have primarily been used as a part of palliative care to alleviate pain, relieve nausea and stimulate appetite. In addition, numerous cell culture and animal studies showed antitumor effects of cannabinoids in various cancer types. Here we reviewed the literature on anticancer effects of plant-derived and synthetic cannabinoids, to better understand their mechanisms of action and role in cancer treatment. We also reviewed the current legislative updates on the use of cannabinoids for medical and therapeutic purposes, primarily in the EU countries. <em>In vitro</em> and <em>in vivo</em> cancer models show that cannabinoids can effectively modulate tumor growth, however, the antitumor effects appear to be largely dependent on cancer type and drug dose/concentration. Understanding how cannabinoids are able to regulate essential cellular processes involved in tumorigenesis, such as progression through the cell cycle, cell proliferation and cell death, as well as the interactions between cannabinoids and the immune system, are crucial for improving existing and developing new therapeutic approaches for cancer patients. The national legislation of the EU Member States defines the legal boundaries of permissible use of cannabinoids for medical and therapeutic purposes, however, these legislative guidelines may not be aligned with the current scientific knowledge.</p> 2019-02-12T00:00:00+01:00 ##submission.copyrightStatement##