The flavonoid fisetin ameliorates renal fibrosis by inhibiting SMAD3 phosphorylation, oxidative damage, and inflammation in ureteral obstructed kidney in mice

Renal fibrosis is characterized by the accumulation of extracellular matrix and inflammatory cells and kidney dysfunction, which is a major pathway in the progression of chronic kidney disease (CKD). Accumulating evidence indicates that oxidative stress plays a critical role in the initiation and progression of CKD via proinflammatory and profibrotic signaling pathways. Fisetin (3,3′,4′,7-tetrahydroxyflavone) has biological activities including antioxidant, anti-inflammatory, and anti-aging effects. Therefore, we evaluated the antifibrotic effects of fisetin on unilateral ureteral obstruction (UUO)-induced kidneys. Methods: C57BL/6 female mice were subjected to right UUO and intraperitoneally injected every other day with fisetin (25 mg/kg/ day) or vehicle from 1 hour before surgery to 7 days after surgery. Kidney samples were analyzed for renal fibrosis (α-smooth muscle actin [α-SMA] expression, collagen deposition, and transforming growth factor [TGF] β1/SMAD3 signaling pathway), oxidative damage (4-HNE and 8-OHdG expression), inflammation (proinflammatory cytokine/chemokine, macrophage, and neutrophil infiltration), and apoptosis (TUNEL staining). Cultured human proximal tubule cells were treated with fisetin before TGF-β to confirm the TGF-β downstream pathway (SMAD2/3 phosphorylation). Results: We found that fisetin treatment protected against renal fibrosis by inhibiting the phosphorylation of SMAD3, oxidative damage, inflammation, apoptotic cell death, and accumulation of profibrotic M2 macrophages in the obstructed kidneys. In cultured human proximal tubular cells, fisetin treatment inhibited TGF-β1–induced phosphorylation of SMAD3 and SMAD2. Conclusion: Fisetin alleviates kidney fibrosis to protect against UUO-induced renal fibrosis, and could be a novel therapeutic drug for obstructive nephropathy.

Diminazene aceturate exacerbates renal fibrosis after unilateral ureteral obstruction in female mice

Diminazene aceturate (DIZE), an angiotensin-converting enzyme 2 (ACE2) activator, exerts anti-inflammatory and antifibrotic effects in a variety of human chronic diseases. However, the role of DIZE in kidney fibrosis and the underlying mechanism remain unclear. Therefore, we investigated the effects of DIZE on the progression of renal fibrosis after unilateral ureteral obstruction (UUO), a well-established model of chronic kidney disease. Methods: C57BL/6 female or male mice were subjected to right UUO. Mice received 15 mg/kg DIZE or vehicle (saline) daily. On the 7th day after UUO, kidneys were collected for analysis of renal fibrosis (α-smooth muscle actin, phosphorylated SMAD3, transforming growth factor (TGF)-β, Masson’s trichrome, and Sirius red staining), inflammation (macrophage infiltration, proinflammatory cytokines/ chemokines), apoptosisecrotic cell death (TUNEL and periodic acid-Schiff staining), and ACE2 activity and messenger RNA (mRNA) expression. Results: Treatment with DIZE exacerbated renal fibrosis by upregulating the profibrotic TGF-β/SMAD3 pathway, proinflammatory cytokine/chemokines (interleukin [IL]-1β, monocyte chemoattractant protein-1, IL-6, and macrophage inflammatory protein-2) levels, M2 macrophage accumulation (CD206, IL-4, IL-10, and CX3CL1), and apoptoticecrotic cell death in the obstructed kidneys of female mice but not male mice. However, DIZE treatment had no effect on ACE2 activity or mRNA expression. Conclusion: DIZE exacerbates UUO-induced renal fibrosis by aggravating tubular damage, apoptosis, and inflammation through independent of angiotensin (1–7), angiotensin II levels, and ACE2 expression/activity, rather than protecting against renal fibrosis after UUO. DIZE also has powerful effects on recruiting macrophages, including the M2-polarized subtype, in female UUO mice.

Yes-associated protein 1 as a prognostic biomarker and its correlation with telomerase in various cancers

Objectives@#The aims of this study were to investigate the expression of Yes-associated protein 1 (YAP1), its prognostic significance, and the correlation between YAP1 and telomerase in various cancers. @*Methods@#The Gene Expression Profiling Interactive Analysis database was used to analyze RNA sequencing data and the survival rate of patients with various cancers in The Cancer Genome Atlas (TCGA) database. PrognoScan was used to analyze the prognostic value of YAP1 expression in various cancers. Tumor Immune Estimation Resource was used to determine the correlation between YAP1 expression and telomerase in various cancer types based on TCGA data. @*Results@#The analysis suggested that YAP1 was differentially expressed between tissues of various cancers and non-tumor tissues. High YAP1 expression was also related to a poor prognosis in adrenocortical carcinoma, bladder urothelial carcinoma, and pancreatic adenocarcinoma. Moreover, YAP1 expression was correlated with the expression of telomerase reverse transcriptase and telomerase RNA component in various cancer types. @*Conclusion@#These results suggest that YAP1 is a potential biomarker with prognostic significance and relevance for oncogene research in various cancer types. The correlation between the expression of YAP1 and telomere-associated genes will help to understand their cancer-promoting mechanisms and interactions.

Analysis of the Cancer Genome Atlas Data to Determine the Prognostic Value of GABPB1L and TERT in Glioblastoma / 계명의대학술지

Glioblastoma multiforme (GBM) is the most lethal type of brain tumor and its prognosis was extremely poor. Here, we studied the clinical and prognostic value of telomerase reverse transcriptase (TERT) and GA Binding Protein Transcription Factor Subunit Beta 1 longer (GABPB1L) mRNA expression in GBM by using open big data. In total 152 GBM patients, gene expressions were downloaded from the Cancer Genome Atlas (TCGA) data and its values were statistically analyzed. TCGA data showed that GABPB1L mRNA expression levels did not correlate with those of TERT mRNA (r = -0.027, p = 0.744). GABPB1L and TERT expressions in GBM were associated with subtype by gene expression. GABPB1L (562.7 ± 76.8 days vs. 479.1 ± 5.8 days, p = 0.401) and TERT (468.1 ± 43.6 days vs. 565.3 ± 75.8 days, p = 0.403) expression was not associated with GBM prognosis. However, disease-free survival was tended to be different in GBM patients according to TERT and GABPB1L expressions though it did not get statistical significance (2122.6 ± 160.2 days vs. 1381.1 ± 244.0 days, p = 0.072). These genes may contribute to the GBM pathogenesis and its further study should be performed in GBM patients and cell lines.

Effect of Bingo for Anatomy Education / 계명의대학술지

Anatomy is knowledge about structure and function of human body. However, with on medical course the vast amount of content, many students have difficulties and burdens about anatomy. The purpose of this study was to investigate the effect of bingo game to improve understanding and learning effect of anatomy. This study was carried out in September, 2018, at 53 medical students. After conducting the bingo, the students were asked about the usefulness, grade reflection, concern, continuation, necessity, appropriate time and interest of bingo. And the relationship between bingo results and grades was also investigated. A total of seven bingo games were with an average of 3.6 bingo (0-11). There was a positive response to the usefulness (3.74 ± 0.92), interest (3.94 ± 0.82), continuation (3.55 ± 1.10), necessity (3.51 ± 1.01), and concern (3.72 ± 1.01) of bingo. However, the appropriateness of the time was neutral (3.26 ± 1.02), and there were negative opinions about grade reflection of bingo (1.40 ± 0.86). The grade of anatomy subject showed a positive correlation with the results of the bingo, but did not reach statistical significance (r = 207, p = 0.088). In addition to anatomy, histology, microbiology, and pharmacology were also considered to be useful educational methods for bingo. Bingo may be useful in medical school curriculum, especially in Anatomy. More research is needed to improve learning efficiency through bingo.

Effect of Bingo for Anatomy Education / 계명의대학술지

Anatomy is knowledge about structure and function of human body. However, with on medical course the vast amount of content, many students have difficulties and burdens about anatomy. The purpose of this study was to investigate the effect of bingo game to improve understanding and learning effect of anatomy. This study was carried out in September, 2018, at 53 medical students. After conducting the bingo, the students were asked about the usefulness, grade reflection, concern, continuation, necessity, appropriate time and interest of bingo. And the relationship between bingo results and grades was also investigated. A total of seven bingo games were with an average of 3.6 bingo (0-11). There was a positive response to the usefulness (3.74 ± 0.92), interest (3.94 ± 0.82), continuation (3.55 ± 1.10), necessity (3.51 ± 1.01), and concern (3.72 ± 1.01) of bingo. However, the appropriateness of the time was neutral (3.26 ± 1.02), and there were negative opinions about grade reflection of bingo (1.40 ± 0.86). The grade of anatomy subject showed a positive correlation with the results of the bingo, but did not reach statistical significance (r = 207, p = 0.088). In addition to anatomy, histology, microbiology, and pharmacology were also considered to be useful educational methods for bingo. Bingo may be useful in medical school curriculum, especially in Anatomy. More research is needed to improve learning efficiency through bingo.

Evaluation of Seeplex(TM) Pneumobacter Multiplex PCR Kit for the Detection of Respiratory Bacterial Pathogens in Pediatric Patients / 대한진단검사의학회지

BACKGROUND: Rapid identification of the causative agent among potential bacterial and viral pathogens is important for the management of acute respiratory disease. In this study, we evaluated the analytical performance and clinical usefulness of a recently-introduced multiplex PCR assay, Seeplex(TM)Pneumobacter detection kit (Seegene Inc., Korea) for the identification of respiratory bacterial pathogens. METHODS: One hundred and eighty one nasopharyngeal aspirates were collected from pediatric patients with respiratory symptoms and analysed by multiplex PCR for the detection of Streptococcus pneumoniae (S.P), Haemophilus influenzae (H.I), Mycoplasma pneumoniae (M.P), Chlamydophila pneumoniae (C.P), Bordetella pertussis (B.P) and Legionella pneumophila (L.P). A comparison of multiplex PCR with conventional culture for the isolation of S.P and H.I was performed on 112 specimens. The cross reactivity of multiplex PCR was also evaluated. RESULTS: Of 181 cases, 81 cases were positive by multiplex PCR (44.8%): 52 cases for S.P (28.7%), 47 cases for H.I (26.0%), 9 cases for M.P (5.0%), 3 cases for B.P (1.7%) and 1 case for C.P (0.6%) including multiple infection cases. The agreement rates between multiplex PCR and culture for S.P and H.I were 92.9% (kappa index=0.84, P<0.001) and 91.1% (kappa index=0.75, P<0.001), respectively. There was no cross reactivity with common bacterial and viral pathogens. CONCLUSIONS: Seeplex(TM) Pneumobacter detection kit could be a useful screening tool for the rapid detection of respiratory bacterial pathogens. Further studies with lower respiratory tract specimens would be needed for the clinical evaluation of S. pneumoniae and H. influenzae detected by multiplex PCR.

Evaluation of iQ200 Automated Urine Microscopy Analyzer / 대한진단검사의학회지

BACKGROUND: Microscopic examination of urine sediment is one of the most commonly performed tests in the clinical laboratory. However, manual microscopic sediment examination is labor-intensive, time-consuming and imprecise. In this study, we evaluated the analytical performance and clinical usefulness of a recently introduced image-based automated urinalysis system, Iris iQ200 (Iris Diagnostics, USA). METHODS: We assessed the iQ200 for linearity, precision and carryover rate using patient's samples and quality control materials. On 337 urine samples, urine sediment analyses performed by the iQ200 were compared with manual microscopy results. RESULTS: The iQ200 showed a good linearity (r2>0.99) for all cellular components analyzed. Within-run and total CVs on urine specimens and quality control samples were less than 10% except for within-run CV for the samples with low concentration of the squamous epithelial cells. The carryover rates were 0.21% for RBCs and 1.92% for WBCs. The agreement rates within one grade between the iQ200 and manual microscopy for RBCs, WBCs, and squamous epithelial cells were 93.8%, 94.2% and 96.9%, respectively. CONCLUSIONS: Since the iQ200 showed a reliable analytical performance and good concordance with manual microscopy, it could be useful in the clinical practice as a screening procedure.

Correlation of In-training Examination Score with the Residency Program or the Score of the Board Examination of Laboratory Medicine / 대한진단검사의학회지

BACKGROUND: An in-training examination is given annually to the all laboratory medicine residents of in Korea. The purpose of this study was to evaluate the results of the in-training examinations according to the examinees' postgraduate years, a number of teaching faculty members and hospital beds, and the score of the board examination. METHODS: A total number of examinees during the 5-year period from 2001 to 2005 were 311. All residents took the same in-training examinations given each year irrespective of the postgraduate year (PGY). RESULTS: The scores of in-training examinations increased with advance in the examinees' PGY (P0.05). The correlation coefficients of each PGY scores were from 0.474 to 0.755 (P<0.01). The scores of the 4th PGY were correlated with those of the board examinations (r=0.627, P<0.001). CONCLUSIONS: These results suggest that the scores of the in-training examinations may be a valid measure of knowledge acquired by residents during their training years and provide a useful information for improving the laboratory medicine residency training program.

Relationship of Free Fatty Acid/Albumin Molar Ratio with Indicators of Erythrocyte Injury in Clinical Conditions with Hypoalbuminemia / 대한임상병리학회지

BACKGROUND: Free fatty acids are well known as an energy source. However, theoretically it could be destructive through oxygen free radical chain reactions unless they are bound to albumin in blood. Recently, the toxicity of oxidative agents in several diseases, and additionally the behavior of antioxidants including albumin against this have been suggested. Therefore, we investigated the relationship between free fatty acid/albumin molar ratio and erythrocyte injury in this study. METHODS: Free fatty acid and albumin were analysed in thirty-eight hypoalbuminemia patients and fifty-six healthy controls. To measure the erythrocyte injury, hemoglobin, absolute and relative reticulocyte counts, and lactate dehydrogenase (LD) were also examined. In addition, glutathione peroxidase (GPX) and superoxide dismutase (SOD) were determined in both fifteen patients and fifteen controls, respectively. RESULTS: The albumin levels in study group (3.06+/-0.28 g/dL) were significantly lower than those of control group (4.94+/-0.21 g/dL). The hemoglobin, reticulocyte counts, and LD levels in study group were significantly different from those of control group (P<0.01), but the free fatty acid concentrations showed no difference between two groups. The free fatty acid/albumin molar ratio in study group was significantly higher than control values. In study group, there were significant correlations between the free fatty acid/albumin molar ratio and (a) LD (r=0.43, P< 0.05), (b) relative reticulocyte count (r=0.39, P<0.05), and (c) hemoglobin (r=-0.31, P<0.01), respectively. The GPX and SOD activities in study group were not statistically different from the control values. There was an inverse correlation between albumin and GPX concentrations in study group (r=-0.36, P<0.01). CONCLUSIONS: These results suggest that toxic effect of unbound free fatty acid with decreased albumin activity as antioxidant may be involved in the cellular injury in hypoalbuminemia patients. Further studies for the correlation of free fatty acid/albumin molar ratio with individual antioxidant status are needed.

An Analysis of Nicotine and its Metabolites in Plasma and Urine Samples of Tobacco Smokers and Nonsmokers by High Performance Liquid Chromatographic Method / 대한임상병리학회지

BACKGROUND: Smoking has been suggested to invoke many health problems. Nicotine, one of the effective major components of tobacco smoke, has a half-life less than 2 hours and is oxidized to its major metabolite, cotinine. This study was conducted to establish the measurement system of nicotine and cotinine by high performance liquid chromatographic method (HPLC) and to set reference range in Korean population. METHODS: Fifteen nonsmokers (25-66 years old, 37.9 average) and 30 smokers (22-63 years old, 27.9 average) were investigated. We modified the methods from Kyerenmaten, et al. and from Hariharan, et al. Urine and heparinized plasma samples were pretreated. Pretreated samples were injected into Waters u-Bondapak C18 reverse column (3.9 mm 30 cm) of Waters HPLC system unit with flow rate of 2 mL/min. Absorbance was monitored at 254 nm of wavelength. RESULTS: The retention times of the NNX (nicotine-1'-N-oxide), cotinine, and nicotine peaks were 2.9, 3.7, 5.1 min, respectively, and readily delayed with increase of pH in the mobile phase. Nicotine and cotinine levels in plasma and urine samples by a modified HPLC method showed high linearity from 0 to 1000 ng/mL for both compounds. Intra- and inter-assay coefficients of variation were 7.49% and 6.54%, respectively for nicotine assay and 5.71% and 14.20%, respectively for cotinine assay. The averages and standard deviations for plasma cotinine, nicotine, urine cotinine, and nicotine in nonsmokers (N=15) were 277.8+/-313.9, 0.7+/-2.4, 382.0+/-273.7, and 17.2+/-27.5 ng/mL, respectively, and in smokers (N=30) were 312.9+/-267.1, 26.3+/-50.1, 1,049.2+/-556.2, and 555.7+/-895.1 ng/mL, respectively (P=0.351, 0.009, 0.0026, 0.000004). CONCLUSIONS: A modified HPLC method for nicotine and cotinine measurement showed a high precision and accuracy. Nicotine and cotinine levels in plasma and urine samples of smokers were significantly higher than those of non-smokers, except for plasma cotinine in passive smokers of nonsmoker group. And this method can be used as a routine test for detection of passive smoking and managing of smoking habit. Reference values of nicotine and cotinine measured in Korean nonsmokers and smokers were suggested.

RBC Sorbitol Analysis in Diabetes Mellitus / 대한임상병리학회지

BACKGROUND: Red blood cell (RBC) sorbitol has been implicated in the pathogenesis of organic complications of diabetes mellitus. W8 investigated RBC sorbitol level as an indicator of glucose control or diabetic complications, and also evaluated whether RBC sorbitol/plasma glucose ratio is an indicator of diabetic complications. METHODS: RBC sorbitol levels were measured in 43 healthy persons and 133 diabetes mellitus (DM) patients by enzymatic method. We also tested linearity, inter- and intra- assay precisions. Plasma glucose and Hb Alc were measured by hexokinase method and HPLC, respectively. Hospital records were reviewed. RESULTS: The intra- and inter-assay coefficients of variation of RBC sorbitol test are 8.7% and 28.5%, respectively. Linearity is good. The RBC sorbitol level(3.60+/-1.00 ug/mL) and RBC sorbitol/plasma glucose ratio (2.37+/-0.98%) in diabetic patients are significantly higher than those in normal control (1.69+/-0.43 ug/mL, 1.85+/-0.49 per mill), respectively(p<0.0001). We can't observe correlation between RBC sorbitol and Hb Alc in BM patients, but observe that in non-treatment DM patients. We also observed correlation between Hb Alc and glucose and reverse correlation between RBC sorbitol ratio and Hb Alc. We can't find significant relation between diabetic complications and RBC sorbitol or RBC sorbitol/plasma glucose. CONCLUSIONS: We suggest that the reference range of normal RBC sorbitol level and RBC sorbitol/plasma glucose ratio by enzymatic method are 1.69+/-0.86 ug/mL and 1.85+/- 0.98%,. These Ire significantly different from DM patients and may be useful in diagnosis of DM.

RBC Sorbitol Analysis in Diabetes Mellitus / 대한임상병리학회지

BACKGROUND: Red blood cell (RBC) sorbitol has been implicated in the pathogenesis of organic complications of diabetes mellitus. W8 investigated RBC sorbitol level as an indicator of glucose control or diabetic complications, and also evaluated whether RBC sorbitol/plasma glucose ratio is an indicator of diabetic complications. METHODS: RBC sorbitol levels were measured in 43 healthy persons and 133 diabetes mellitus (DM) patients by enzymatic method. We also tested linearity, inter- and intra- assay precisions. Plasma glucose and Hb Alc were measured by hexokinase method and HPLC, respectively. Hospital records were reviewed. RESULTS: The intra- and inter-assay coefficients of variation of RBC sorbitol test are 8.7% and 28.5%, respectively. Linearity is good. The RBC sorbitol level(3.60+/-1.00 ug/mL) and RBC sorbitol/plasma glucose ratio (2.37+/-0.98%) in diabetic patients are significantly higher than those in normal control (1.69+/-0.43 ug/mL, 1.85+/-0.49 per mill), respectively(p<0.0001). We can't observe correlation between RBC sorbitol and Hb Alc in BM patients, but observe that in non-treatment DM patients. We also observed correlation between Hb Alc and glucose and reverse correlation between RBC sorbitol ratio and Hb Alc. We can't find significant relation between diabetic complications and RBC sorbitol or RBC sorbitol/plasma glucose. CONCLUSIONS: We suggest that the reference range of normal RBC sorbitol level and RBC sorbitol/plasma glucose ratio by enzymatic method are 1.69+/-0.86 ug/mL and 1.85+/- 0.98%,. These Ire significantly different from DM patients and may be useful in diagnosis of DM.

Leukemic conversion from bone marrow necrosis / 대한혈액학회지

No abstract available.