• Title/Summary/Keyword: Real-Time polymerase chain reaction (PCR)

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Detection of Epstein-Barr Virus and Cytomegalovirus in Gastric Cancers in Kerman, Iran

  • Leila, Zaruni;Arabzadeh, Seyed Alimohammad;Afshar, Reza Malekpour;Afshar, Abbas Aghaei;Mollaei, Hamid Reza
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2423-2428
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    • 2016
  • Gastric cancer (GC) is a multifactorial disease with different factors having roles in its genesis. Helicobacter pylori and Epstein-Barr virus (EBV) are known infectious agents that could contribute. In addition, there is evidence of a relationship with cytomegalovirus (CMV). Since data on CMV prevalence in gastric cancer are limited, we here evaluated the frequency of EBV and CMV in Iranian patients. Ninety paraffin blocks of GC tissues from patients in Kerman were evaluated for the presence of EBV and CMV genomes by real-time polymerase chain reaction. EBV was detected in 10 cases (11.1%) and CMV in seven. One out of 17 female patients (5.88%) and nine out of 73 male patients (12.3%) were positive for EBV, while one out of 17 female patients (5.88%) and six out of 73 male patients (8.22%) were positive for CMV. The mean age for EBV-positive patients was $60.5{\pm}14.9years$ and the mean age for CMV-positive patients was $67.9{\pm}12.3years$. This study shows that the frequency of EBV-associated GC is high in Kerman. It also indicates that further studies of associations between GC and CMV are warranted, covering larger samples and populations from different areas of the world.

The Inhibition of MicroRNA-139-5p Promoted Osteoporosis of Bone Marrow-Derived Mesenchymal Stem Cells by Targeting Wnt/Beta-Catenin Signaling Pathway by NOTCH1

  • Feng, Yimiao;Wan, Pengbo;Yin, Linling;Lou, Xintian
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.448-458
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    • 2020
  • We investigated the therapeutic effects of microRNA-139-5p in relation to osteoporosis of bone marrow-derived mesenchymal stem cell (BMSCs) and its underlying mechanisms. In this study we used a dexamethasone-induced in vivo model of osteoporosis and BMSCs were used for the in vitro model. Real-time quantitative polymerase chain reaction (RT-PCR) and gene chip were used to analyze the expression of microRNA-139-5p. In an osteoporosis rat model, the expression of microRNA-139-5p was increased, compared with normal group. Down-regulation of microRNA-139-5p promotes cell proliferation and osteogenic differentiation in BMSCs. Especially, up-regulation of microRNA-139-5p reduced cell proliferation and osteogenic differentiation in BMSCs. Overexpression of miR-139-5p induced Wnt/β-catenin and down-regulated NOTCH1 signaling in BMSCs. Down-regulation of miR-139-5p suppressed Wnt/β-catenin and induced NOTCH1 signaling in BMSCs. The inhibition of NOTCH1 reduced the effects of anti-miR-139-5p on cell proliferation and osteogenic differentiation in BMSCs. Activation of Wnt/β-catenin also inhibited the effects of anti-miR-139-5p on cell proliferation and osteogenic differentiation in BMSCs. Taken together, our results suggested that the inhibition of microRNA-139-5p promotes osteogenic differentiation of BMSCs via targeting Wnt/β-catenin signaling pathway by NOTCH1.

Environmental Investigation of a Long-term Care Hospital with Respect to COVID-19

  • Park, Min Woo;Shin, Seung Hwan;Cha, Jeong Ok;Lim, Hyeon Jeong;Kim, Jun Nyun
    • Journal of Environmental Health Sciences
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    • v.46 no.5
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    • pp.599-609
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    • 2020
  • Objectives: Coronavirus disease 2019 (COVID-19) first emerged in December 2019 in Wuhan, China, and has rapidly become a global pandemic with over 26.4 million confirmed cases and approximately 871,000 fatalities worldwide as of this writing. In the Republic of Korea, disease clusters frequently occurred in long-term care hospitals where the majority of residents are elderly with underlying medical conditions. Despite the fact that public health authorities and local community health centers have put tremendous efforts into preventing the spread of disease, positive cases have continued to occur. Thus, the Korea Centers for Disease Control & Prevention rapid response team decided to conduct an environmental investigation of a long-term care hospital to identify whether environmental contamination has remained and contributed to the spread of COVID-19. Methods: An environmental investigation was conducted at Hospital A. The characteristics of the facility and its HVAC system were assessed by checking the layout and interviewing the people in charge. A total of 64 surface samples were collected from areas of concern, including patient rooms, toilets, elevators, and nurses' station. These samples were tested by a regional health and environmental research institute using real-time reverse transcription polymerase chain reaction. Results: All samples from Hospital A were confirmed to be negative. Through interviews with high-level personnel at the regional community health center, we found that extensive disinfection is frequently performed on potentially contaminated areas in Hospital A in accordance with government guidelines. Conclusion: The environmental control measures implemented in Hospital A had been sufficient for mitigating the risk of further infection, suggesting that such measures may also be effective for other long-term health care facilities.

IL-6-miR-210 Suppresses Regulatory T Cell Function and Promotes Atrial Fibrosis by Targeting Foxp3

  • Chen, YingWei;Chang, GuoDong;Chen, XiaoJie;Li, YunPeng;Li, HaiYu;Cheng, Dong;Tang, Yi;Sang, HaiQiang
    • Molecules and Cells
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    • v.43 no.5
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    • pp.438-447
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    • 2020
  • The aim of this study was to explore the role of IL-6-miR-210 in the regulation of Tregs function and atrial fibrosis in atrial fibrillation (AF). The levels of interleukin (IL)-6 and IL-10 in AF patients were detected by using ELISA. Proportions of Treg cells were detected by fluorescence activated cell sorting analysis in AF patients. The expression of Foxp3, α-SMA, collagen I and collagen III were determined by western blot. The atrial mechanocytes were authenticated by vimentin immunostaining. The expression of miR-210 was performed by quantitative real-time polymerase chain reaction (qRT-PCR). TargetScan was used to predict potential targets of miR-210. The cardiomyocyte transverse sections in AF model group were observed by H&E staining. The myocardial filaments were observed by masson staining. The level of IL-6 was highly increased while the level of IL-10 (Tregs) was significantly decreased in AF patients as compared to normal control subjects, and IL-6 suppressed Tregs function and promoted the expression of α-SMA, collagen I and collagen III. Furthermore, miR-210 regulated Tregs function by targeting Foxp3, and IL-6 promoted expression of miR-210 via regulating hypoxia inducible factor-1α (HIF-1α). IL-6-miR-210 suppresses regulatory T cell function and promotes atrial fibrosis by targeting Foxp3.

A Case of Mycobacterium abscessus Lung Disease in a Patient with H1N1 Influenza Pneumonia (H1N1 Influenza 폐렴 환자에서 발생한 Mycobacterium abscessus 감염 1예)

  • Jung, Kyoung-Won;Yu, Hoon;Lee, Tae-Hoon;Kwon, O-Sung;Choi, Suk-Won;Kang, Se-Hun;Shim, Tae-Sun;Kim, Dong-Soon;Song, Jin-Woo
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.3
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    • pp.207-211
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    • 2010
  • Novel influenza A (H1N1) virus is a common pathogen of febrile respiratory infection recently. Here, we report the case of a 63-year-old male patient who presented with 3 days' ongoing cough and fever. He was diagnosed with novel influenza A (H1N1) pneumonia by real-time reverse-transcriptase-polymerase-chain-reaction (rRT-PCR). During treatment for novel influenza A (H1N1), his symptoms and radiologic findings improved initially, but multiple lung nodules developed subsequently and found on chest x-ray (on the 5th hospital day). Mycobacterium abscessus was isolated repeatedly from sputum and bronchoalveolar lavage fluid. To our knowledge, this is the first reported case of Mycobacterium abscessus lung disease in a patient with H1N1 influenza pneumonia.

Genetic Susceptibility to Esophageal Cancer due to CYP1A1 Gene Variant rs4646903 in Tobacco Addicted Patients of Pashtun Ethnicity: a Case Control Study in Khyber Pakhtunkhwa Province of Pakistan

  • Zakiullah, Zakiullah;Saeed, Muhammad;Ali, Sajid;Javed, Nabila;Khisroon, Muhammad;Muhammad, Basir;Khuda, Fazli;Ahmad, Saeed;Ismail, Mohammad
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6715-6720
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    • 2014
  • The purpose of this study was to evaluate associations of the CYP1A1 gene variant rs4646903 polymorphism with the risk of developing esophageal cancer (EC). A case-control study was carried out in Pashtun population of Khyber Pakhtunkhwa province of Pakistan in which 140 hospital based EC cases and 196 population based healthy controls exposed to similar environmental conditions were included. A specific method based on the real time polymerase chain reaction (RT-PCR) was used to detect genotypes in case and control groups and results were then analyzed with SPSS version 20. In our population, individuals with CC and TC genotypes of the CYP1A1 rs4646903 polymorphism had significantly higher risk of EC (adjusted odds (OR): 15.709, 95%CI: 6.065-40.686, OR: 3.256 95%CI: 1.902-5.574 respectively). The 'C' allele was strongly associated with the disease (p< 0.0001). Adjusted OR was higher (1.5 times in C/C) in case of variant alleles that show the contribution of environmental and nutritional factors towards the development of EC. Our findings suggest that presence of the 'C' allele of rs4646903 (T>C) may be one of the risk alleles for EC susceptibility in Pashtun population.

Galectin-3-independent Down-regulation of GABABR1 due to Treatment with Korean Herbal Extract HAD-B Reduces Proliferation of Human Colon Cancer Cells

  • Kim, Kyung-Hee;Kwon, Yong-Kyun;Cho, Chong-Kwan;Lee, Yeon-Weol;Lee, So-Hyun;Jang, Sang-Geun;Yoo, Byong-Chul;Yoo, Hwa-Seong
    • Journal of Pharmacopuncture
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    • v.15 no.3
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    • pp.19-30
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    • 2012
  • Objectives: Many efforts have shown multi-oncologic roles of galectin-3 for cell proliferation, angiogenesis, and apoptosis. However, the mechanisms by which galectin-3 is involved in cell proliferation are not yet fully understood, especially in human colon cancer cells. Methods: To cluster genes showing positively or negatively correlated expression with galectin-3, we employed human colon cancer cell lines, SNU-61, SNU-81, SNU-769B, SNU-C4 and SNU-C5 in high-throughput gene expression profiling. Gene and protein expression levels were determined by using real-time quantitative polymerase chain reaction (PCR) and western blot analysis, respectively. The proliferation rate of human colon cancer cells was measured by using a 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Results: Expression of ${\gamma}$-aminobutyric acid B receptor 1 (GABABR1) showed a positive correlation with galectin-3 at both the transcriptional and the translational levels. Down-regulation of galectin-3 decreased not only GABABR1 expression but also the proliferation rate of human colon cancer cells. However, Korean herbal extract, HangAmDan-B (HAD-B), decreased expression of GABABR1 without any expressional change of galectin-3, and offset ${\gamma}$-aminobutyric acid (GABA)-enhanced human colon cancer cell proliferation. Conclusions: Our present study confirmed that GABABR1 expression was regulated by galectin-3. HAD-B induced galectin-3-independent down-regulation of GABABR1, which resulted in a decreased proliferation of human colon cancer cells. The therapeutic effect of HAD-B for the treatment of human colon cancer needs to be further validated.

Relationship Between Mitochondrial DNA Copy Number, Metabolic Abnormalities and Hepatic Steatosis (지방간 및 대사 인자들과 말초혈액 백혈구의 사립체 DNA copy 수와의 연관성)

  • Kwon, Kil-Young;Jun, Dae-Won
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.6
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    • pp.2093-2098
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    • 2010
  • Insulin resistance plays a central role in fatty liver, a part of the metabolic syndrome. This study examined the relationship between fatty liver, metabolic abnormalities and mitochondrial DNA [mtDNA] copy number in peripheral blood that is correlated with diabetes or metabolic markers. Fatty liver was assessed by questionnaire on alcohol consumption and abdominal ultrasonography. MtDNA copy number in peripheral leukocytes was measured by a real-time quantitative polymerase chain reaction [PCR]. Among 445 subjects, 148 subjects had hepatic steatosis and 297 were controls. mtDNA copy number was significantly lower in fatty liver group in comparison with that of normal finding group. This result is similar in both groups, alcoholic or non-alcoholic fatty liver group. MtDNA copy number was inversely correlated with alanine aminotransferase [ALT], aspartate aminotransferase [AST], gamma-glutamyltransferase [$\gamma$-GTP], body mass index [BMI], waist circumference, diastolic blood pressure, and free fatty acid. MtDNA copy number in peripheral leukocytes was associated with fatty liver and insulin resistance related factors.

Anti-Oxidant Activity and Anti-Inflammatory Effects of Spiraea fritschiana Schneid Extract (참조팝나무 추출물의 항산화 활성 및 항염증 효과)

  • Choi, Eun Yeong;Heo, Seong Il;Kwon, Yong Soo;Kim, Myong Jo
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.1
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    • pp.31-37
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    • 2016
  • Background : We studied the anti-oxidant activity and anti-inflammatory effects of Spiraea fritschiana Schneid extract (SFSE). Methods and Results : The SFSE was prepared using methanol and was evaluated for its total phenol and flavonoid content, DPPH (1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity, reducing power, and effect on nitric oxide (NO) production, and cell viability by using real-time polymerase chain reaction (PCR). The total phenol content was $212.78{\mu}g{\cdot}galli$c acid equivalent (GAE)/mg and the total flavonoid content was $66.84{\mu}g{\cdot}quercetin$ equivalent (QE)/mg. The extract showed antioxidant activity (DPPH free-radical scavenging activity) with $RC_{50}$ value of $76.61{\mu}g/m{\ell}$. The reducing power of the extract was Abs 0.58 at $250{\mu}g/m{\ell}$. Cell viability was determined using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. To evaluate anti-inflammatory activity, we examined the inhibitory effects on lipopolysaccharide-(LPS)-induced NO production in RAW 264.7 cells. The NO inhibition rate was 90% at $200{\mu}g/m{\ell}$ SFSE. At the same concentration, the expression of pro-inflammatory genes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 also decreased. Conclusions : Our results suggest that SFSE is a novel resource for the development of foods and drugs that possess anti-oxidant and anti-inflammatory activity.

Expression of Various Pattern Recognition Receptors in Gingival Epithelial Cells

  • Shin, Ji-Eun;Ji, Suk;Choi, Young-Nim
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.77-82
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    • 2008
  • Innate immune response is initiated by the recognition of unique microbial molecular patterns through pattern recognition receptors (PRRs). The purpose of this study is to dissect the expression of various PRRs in gingival epithelial cells of differentiated versus undifferentiated states. Differentiation of immortalized human gingival epithelial HOK-16B cells was induced by culture in the presence of high $Ca^{2+}$ at increased cell density. The expression levels of various PRRs in HOK-16B cells were examined by realtime reverse transcription polymerase chain reaction (RTPCR) and flow cytometry. In addition, the expression of human beta defensins (HBDs) was examined by real time RT-PCR and the amounts of secreted cytokines were measured by enzyme linked immunosorbent assay. In undifferentiated HOK-16B cells, NACHT-LRR-PYDcontaining protein (NALP) 2 was expressed most abundantly, and toll like receptor (TLR) 2, TLR4, nucleotide-binding oligomerization domain (NOD) 1, and NOD2 were expressed in substantial levels. However, TLR3, TLR7, TLR8, TLR9, ICE protease-activating factor (IPAF), and NALP6 were hardly expressed. In differentiated cells, the levels of NOD2, NALP2, and TLR4 were different from those in undifferentiated cells at RNA but not at protein levels. Interestingly, differentiated cells expressed the increased levels of HBD-1 and -3 but secreted reduced amount of IL-8. In conclusion, the repertoire of PRRs expressed by gingival epithelial cells is limited, and undifferentiated and differentiated cells express similar levels of PRRs.