• Title/Summary/Keyword: PCR (Polymerase Chain Reaction)

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Association Study between Serotonin 1A Receptor Gene rs6295 Polymorphism and Tardive Dyskinesia in Patients with Schizophrenia (조현병 환자에서의 지연성 운동이상 발생과 세로토닌 1A 수용체유전자 rs6295 다형성과의 연관성)

  • Lee, Youn-Jung;Namgoong, Yoon;Cho, Areum;Lee, Heon-Jeong
    • Korean Journal of Schizophrenia Research
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    • v.23 no.2
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    • pp.71-77
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    • 2020
  • Objectives: Tardive dyskinesia (TD) is a movement disorder that is characterized by hyperkinetic movements. Previous studies have suggested that the serotonergic systems are correlated with TD vulnerability. In this study, the association between a single-nucleotide polymorphism (SNP) of the serotonin 1A receptor gene (HTR1A) rs6295 and TD was investigated. Methods: We investigated whether HTR1A rs6295 SNP is associated with antipsychotic-induced TD in 280 Korean patients with schizophrenia. Patients with schizophrenia having TD (n=105) and those without TD (n=175) were matched for their antipsychotic exposures and other relevant variables. The HTR1A rs6295 SNP was analyzed using polymerase chain reaction (PCR)-based methods. Results: There was no significant difference in the distribution of genotypic (χ2=2.70, p=0.26) and allelic (χ2=1.87, p=0.17) frequencies between the patient groups with TD and without TD. There was no significant difference in total abnormal involuntary movement scale score (F=0.39, p=0.68) among the genotype group either. Conclusion: Although there were no differences in genotypic and allelic frequency between patient groups with and without TD, further studies on association of TD with other SNPs of HTRA1 are needed to understand the pathophysiological mechanism of TD.

Distribution and genetic diversity of Feline calicivirus in Moscow metropolitan area

  • Alina, Komina;Nikita, Krasnikov;Oksana, Kucheruk;Elena, Zhukova;Anton, Yuzhakov;Alexey, Gulyukin
    • Journal of Veterinary Science
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    • v.23 no.6
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    • pp.92.1-92.8
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    • 2022
  • Background: Feline calicivirus (FCV) is widespread throughout the world. An FCV infection is associated with conjunctivitis, rhinitis, and mouth ulcers that can lead to the animal's death. Because vaccination is not always effective, it is necessary to monitor the infection regularly. Objectives: This study examined the FCV epizootic situation in the Moscow metropolitan area by conducting a molecular phylogenetic analysis of the virus isolates. Methods: Samples from 6213 animals were examined by a reverse transcription polymerase chain reaction. For phylogenetic analysis, 12 nucleotide sequences obtained from animal samples were selected. Sequencing was performed using the Sanger method. Phylogenetic analysis was conducted using the Maximum Likelihood method. Results: The FCV genome was detected in 1,596 (25.7%) samples out of 6,213. In 2018, calicivirus was detected in 18.9% of samples, 27.8% in 2019, 21.4% in 2020, and 32.6% in 2021. Phylogenetic analysis of the F ORF2 region and the ORF3 start region led to division into two FCV genogroups. Most of the isolates (8 out of 12) were close to the Chinese strains. On the other hand, there were isolates closely related to European and American strains. The isolates circulating in Moscow were not included in clusters with vaccine strains; their nucleotide similarity varied from 77% to 83%. Conclusions: This study revealed a high prevalence and genetic diversity of the FCV in Moscow. The epizootic situation remains stably tense because 24 viruses were detected in 25% of animals annually.

A Rapid and Simple Method for DNA Preparation of Magnaporthe oryzae from Single Rice Blast Lesions for PCR-Based Molecular Analysis

  • Liying, Dong;Shufang, Liu;Jing, Li;Didier, Tharreau;Pei, Liu;Dayun, Tao;Qinzhong, Yang
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.679-684
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    • 2022
  • Rice blast is one of the most destructive diseases of rice worldwide, and the causative agent is the filamentous ascomycete Magnaporthe oryzae. With the successful cloning of more and more avirulence genes from M. oryzae, the direct extraction of M. oryzae genomic DNA from infected rice tissue would be useful alternative for rapid monitoring of changes of avirulence genes without isolation and cultivation of the pathogen. In this study, a fast, low-cost and reliable method for DNA preparation of M. oryzae from a small piece of infected single rice leaf or neck lesion was established. This single step method only required 10 min for DNA preparation and conventional chemical reagents commonly found in the laboratory. The AvrPik and AvrPi9 genes were successfully amplified with the prepared DNA. The expected DNA fragments from 570 bp to 1,139 bp could be amplified even three months after DNA preparation. This method was also suitable for DNA preparation from M. oryzae strains stored on the filter paper. All together these results indicate that the DNA preparation method established in this study is reliable, and could meet the basic needs for polymerase chain reaction-based analysis of M. oryzae.

Comparative Study of Bojungikgitang in Korea, Japan and China on the Anti-Inflammatory and Anti-Oxidative Effects (보중익기탕 (補中益氣湯)의 한국, 중국, 일본 처방에 대한 항염증 및 항산화 효과 비교 연구)

  • Choi, Hye-Min;Kim, Hui-Hun;Lee, Hwa-Dong
    • The Korea Journal of Herbology
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    • v.29 no.1
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    • pp.53-60
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    • 2014
  • Objectives : Bojungikgitang (BJT), the Oriental medical prescription has been traditionally used about improvement of immune response and infective disease at Asian nation. In this study, we has compared about the anti-inflammatory and antioxidative effects on BJT of three countries including Korea (Korean Traditional Medicine, KTM), China (Traditional Chinese Medicine, CTM) and Japan (Japanese Traditional Medicine, JTM). Methods : We has basically using LPS-stimulated RAW 264.7 cells. The expression of these inflammatory mediators has measured using enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR). Also, free radical scavenging assay has tested for anti-oxidative activity as well as the contents of total flavonoid and polyphenol. Results : As a result, we were founded the inhibitory effects of BJT (KTM, CTM, JTM) on LPS-induced production of NO, TNF-${\alpha}$ and IL-6 as well as the anti-oxidative activities. Especially the KTM was most effective in anti-inflammatory and anti-oxidative activities. Conclusions : These results indicate that BJT (KTM, CTM, JTM) has a good anti- inflammatory and anti-oxidative effects. But, there were degree of effects on between pharmacopoeia of the countries. Thus, further study is required that find appropriate methods for extracting as well as establish of standardized processes in order to improve the quality of BJT (KTM, CTM, JTM) as an anti-inflammatory and anti-oxidative agent for treatment of inflammatory diseases.

Effects of Baicalein on Neuroinflammation in Lipopolysaccharide-treated Mice (Baicalein이 Lipopolysaccharide에 의한 생쥐의 Neuroinflammation에 미치는 영향)

  • Ha, Gyung-Woon;Kim, Youn-Sub
    • The Korea Journal of Herbology
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    • v.28 no.2
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    • pp.93-101
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    • 2013
  • Objects : Baicalein is a major bioactive flavonoid component of Scutellaria baicalensis Georgi that shows a wide range of biological activities, including neuroprotections and anti-inflammatory actions. Hence it is a potential therapeutic material for the treatment of neuroinflammation. In this study, we investigated the modulatory effect of baicalein on neuroinflammation. Method : Pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 mRNA), COX-2 mRNA expression and microglial activation in the brain tissue is induced by systemic lipopolysaccharide (LPS) treatment in C57BL/6 mice. Baicalein was treated orally with 10, 20, and 30 mg/kg 1 hour prior to the LPS (3 mg/kg, i.p.) injection. TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and COX-2 mRNA expression in the brain tissue was measured by the quantitative real-time polymerase chain reaction(PCR) method. Iba1 expression in the brain was measured by western blotting method. Microglia was observed with immunohistochemistry. Results : Baicalein 30 mg/kg significantly attenuated the expression of TNF-${\alpha}$, IL-$1{\beta}$, IL-6 and COX-2 mRNA in the brain tissue. Baicalein 20 mg/kg significantly attenuated the expression of IL-6 mRNA in the brain tissue. Baicalein 30 mg/kg significantly attenuated the expression of Iba1 protein expression in the brain tissue. Baicalein 30 mg/kg significantly decreased the number and cell size of microglia in the cerebral cortex and hypothalamic region and the area percentage of Iba1-expressed microglia in the hippocampus. Conclusion : These results demonstrated that baicalein attenuates LPS induced neuroinflammation in the mice via reduction of pro-inflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, IL-6), COX-2 mRNA expression and microglial activation.

Protective effects of Smilacis Glabrae Rhizoma on acute pancreatitis (토복령(土茯苓) 물 추출물의 급성 췌장염 보호 효과)

  • Kim, Myoung-Jin;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.33 no.2
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    • pp.45-52
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    • 2018
  • Objectives : Smilacis Glabrae Rhizoma (SGR) has been used in traditional medicine for treating diseases related to inflammation and cancer. However, the protective effect of SGR has not been reported in the study of acute pancreatitis (AP). Therefore, this study was performed to evaluate the protective effects of SGR water extract on cerulein-induced AP. Methods : AP was induced by intraperitoneal injection of supramaximal concentrations of stable cholecystokinin analogue cerulein ($50{\mu}g/kg$) 6 times at intervals of one hour in mice. SGR water extract (0.01, 0.05, or 0.1 g/kg) was administrated intraperitoneally 1 h prior to the first injection of cerulein. The mice were sacrificed at 6 h after the final injection of cerulein. Pancreas and lung were rapidly removed for histochemical examination and myeloperoxidase (MPO) assay. In addition, polymerase chain reaction (PCR) was performed to examine mRNA levels of proinflammatory cytokines such as interleukin(IL)-6, $IL-1{\beta}$ and Tumor necrosis $factor(TNF)-{\alpha}$. Results : Administration of SGR water extract significantly inhibited pancreatic weight to body weight ratio. In addition, SGR treatment inhibited the histological damages and increase of MPO activity in both pancreas and lung during AP. Also, mRNA levels of IL-6 but not $IL-1{\beta}$and $TNF-{\alpha}$ were inhibited by SGR water extract against AP. Conclusion : Our results revealed that pre-treatment of SGR water extract reduces the severity of cerulein-induced AP. Therefore, this study suggest that SGR could be used as a drug or agent to prevent AP.

Hepatoprotecive Effects of Puerariae Flos Extract on Experimental Non-alcoholic Fatty Liver Disease Models (비알콜성 지방간 동물모델에서 갈화추출물의 간보호 효과)

  • Hwang, Hyeong-Chil;Kong, Ryong;Kang, Ok-Hwa;Kwo, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.1-8
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    • 2018
  • Objectives : The aim of this study is to investigate the preventive effect of Puerariae Flos ethanol extract (PE) on methionin and choline deficient (MCD)-diet-induced Nonalcoholic fatty liver disease (NAFLD) in C57BL/6J mice. Methods : In the in vivo experiments, C57BL/6J mice were divided into 4 groups; Normal group, Control group, MCD+PE 100 group, and MCD+PE 300 group. After 4 weeks, body weight, liver weight, biochemical parameters for liver function test, histological changes, reverse transcription polymerase chain reaction (RT-PCR), and Western blot were assessed. Results : Mice lost body weight with the MCD-diet and the MCD+PE 100 group and MCD+PE 300 groups lost less than the control group, though showed no statistical significance. Liver weights were decreased by the MCD diet, but MCD+PE 300 groups were increased significantly. In the liver function test, all the values were decreased with the MCD-diet, MCD+PE 100 group and MCD+PE 300 groups were increased significance. In histological findings of the livers, MCD-diet induced severe fatty accumulation in the livers, but this fatty change was reduced in the MCD+PE 100 group and MCD+PE 300 groups was inhibited respectively. In lipid accumulation factors (such as SREBP-1c, $C/EBP{\alpha}$, PPAR-${\gamma}$), MCD+PE 100 group and MCD+PE 300 groups showed inhibitory effect on liver lipogenesis by reducing associated gene expressions caused by MCD diet. Conclusions : We were able to know that Puerariae Flos ethanol extract (PE) shown hepatoprotective effects via a decrease on the hepatic lipogenesis factors in the experimental NAFLD Models.

Porcine parvovirus nonstructural protein NS1 activates NF-κB and it involves TLR2 signaling pathway

  • Jin, Xiaohui;Yuan, Yixin;Zhang, Chi;Zhou, Yong;Song, Yue;Wei, Zhanyong;Zhang, Gaiping
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.50.1-50.16
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    • 2020
  • Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions: These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

Inhibitory Effects of GGX on Lung Injury of Chronic Obstructive Lung Disease (COPD) Mice Model (만성폐쇄성폐질환 동물모델에서 GGX의 폐손상 억제 효과)

  • Kim, Tae Hyeon;Yang, Won Kyung;Lee, Su Won;Kim, Seung Hyung;Lyu, Yee Ran;Park, Yang Chun
    • The Journal of Korean Medicine
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    • v.42 no.3
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    • pp.56-71
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    • 2021
  • Objectives: This study is aimed to evaluate the protective effects of GGX on lung injury of Chronic Obstructive Lung Disease (COPD) mice model. Materials and Methods: C57BL/6 mice were challenged with lipopolysaccharide (LPS) and cigarette smoke extract (CSE) and then treated with vehicle only (Control group), dexamethasone 3 mg/kg (Dexa group), gam-gil-tang 200 mg/kg (GGT group), GGX 100, 200, and 400 mg/kg (GGX group). After sacrifice, its bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed with cytospin, Enzyme-Linked Immunosorbent Assay (ELISA), real-time polymerase chain reaction (PCR) and hematoxylin & eosin (H&E), and Masson's trichrome staining. Results: In the COPD model, GGX significantly inhibited the increase of neutrophils, TNF-𝛼, IL-17A, CXCL-1, MIP2 in BALF and TNF-𝛼, IL-1𝛽, IL-10 mRNA expression in lung tissue. It also decreased the severity of histological lung injury. Conclusion: This study suggests the usability of GGX for COPD patients by controlling lung tissue injury.

COVID-19 Diagnosis from CXR images through pre-trained Deep Visual Embeddings

  • Khalid, Shahzaib;Syed, Muhammad Shehram Shah;Saba, Erum;Pirzada, Nasrullah
    • International Journal of Computer Science & Network Security
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    • v.22 no.5
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    • pp.175-181
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    • 2022
  • COVID-19 is an acute respiratory syndrome that affects the host's breathing and respiratory system. The novel disease's first case was reported in 2019 and has created a state of emergency in the whole world and declared a global pandemic within months after the first case. The disease created elements of socioeconomic crisis globally. The emergency has made it imperative for professionals to take the necessary measures to make early diagnoses of the disease. The conventional diagnosis for COVID-19 is through Polymerase Chain Reaction (PCR) testing. However, in a lot of rural societies, these tests are not available or take a lot of time to provide results. Hence, we propose a COVID-19 classification system by means of machine learning and transfer learning models. The proposed approach identifies individuals with COVID-19 and distinguishes them from those who are healthy with the help of Deep Visual Embeddings (DVE). Five state-of-the-art models: VGG-19, ResNet50, Inceptionv3, MobileNetv3, and EfficientNetB7, were used in this study along with five different pooling schemes to perform deep feature extraction. In addition, the features are normalized using standard scaling, and 4-fold cross-validation is used to validate the performance over multiple versions of the validation data. The best results of 88.86% UAR, 88.27% Specificity, 89.44% Sensitivity, 88.62% Accuracy, 89.06% Precision, and 87.52% F1-score were obtained using ResNet-50 with Average Pooling and Logistic regression with class weight as the classifier.