1.Effect of miR-21 on migration and invasion ability in human laryngeal squamous carcinoma cell Hep 2
Yongjun LIU ; Yanfei GUAN ; Shun CHANG
Chongqing Medicine 2016;45(21):2906-2908,2913
Objective To explore the effect of microRNA‐21(miR‐21) on the migration and invasion ability in human laryn‐geal squamous carcinoma cell Hep2 .Methods The MTT method was used to detect the viability of Hep2 cells at 48 h after miR‐21 inhibitor and miR‐21 NC transferring into Hep2 cells by LipofectamineTM 2000 .The cell migration ability was detected by using the scratch test .The cell invasion ability was detected by using the Transwell method .The activation of phosphatase and tensin homo‐logue deleted on chromosome 10 (PTEN)/phosphatidylinositol 3 kinase (PI3K) /protein kinase B(Akt) signal pathway and the expression of matrix metalloproteinase 2 (MMP2) ,MMP9 ,reversion inducing cysteine rich protein with kazal motif (RECK) was detected by using the Western blotting .Results Compared with miR‐21 NC ,miR‐21 inhibitor could significantly reduce the Hep2 cellviability[(0.688±0.043)vs.(0.375±0.012)],inhibitedthemigrationability[(6.57±0.02)μm vs.(20.49±2.18)μm]and invasion ability[(100 .7 ± 10 .2) vs .(46 .8 ± 4 .3)] ,and the differences were statistically significant (P<0 .01) ,meanwhile miR‐21 inhibitor could down‐regulate the expression of PI3K ,MMP2 and MMP9(P<0 .01) ,and reduced the phosphorylation level of Akt (P<0 .01) ,up‐regulated the expression of PTEN and RECK (P<0 .01) .Conclusion miR‐21 inhibitor can significantly suppress the migration and invasion ability of Hep2 ,which may be related with the PTEN/PI3K/Akt signal pathway .
2.Relationship between expression of PD-L1 and microsatellite instability of colorectal cancer and its significance
Wen FENG ; Xinxin WU ; Yongjun GUO ; Yuxi CHANG ; Qingxin XIA
Chinese Journal of Clinical Oncology 2017;44(12):589-593
Objective:To investigate the expression and clinical significance of PD-L1 in colorectal cancer (CRC). Methods:A total of 210 CRC patients who accepted radical surgery in our hospital from January 2015 to January 2016 were divided into three groups, namely, high-frequency microsatellite instability (MSI-H), low-frequency microsatellite instability (MSI-L), and microsatellite stable (MSS). The expression of PD-L1 was detected by immunohistochemistry, and the expression characteristics of PD-L1 in different types of CRC were analyzed. Results:CRC cases with low differentiation had a higher expression of PD-L1 than CRC patients with high differ-entiation (P<0.05). PD-L1 had a positive rate of 75.8%in the MSI-H group and a rate of 9.3%in the MSI-L and MSS groups, wherein the difference between the two groups was statistically significant (P<0.05). Conclusion:PD-L1 was positively expressed in some CRC tu-mor tissues, and its positive rate was significantly higher in MSI-H than in MSI-L and MSS. The therapeutic effect of a PD-L1 blocker for patients with MSI-H CRC might be preferable.
3.Preliminary study of estrogen effects on calcium free smooth muscle cells at the endometrialmyometrial interface in uteri with adenomyosis
Liping WANG ; Sha WANG ; Ying ZHANG ; Yongjun WANG ; Henghui ZHANG ; Yanan CHANG ; Guoli LI ; Hua DUAN
Chinese Journal of Obstetrics and Gynecology 2012;47(5):351-354
Objective To investigate the effect and mechanism of estrodial (E2) on intracellular free calcium in the endometrial-myometrial interface (EMI) smooth muscle cells from uteri with adenomyosis.MethodsFrom March 2011 to October 2011,16 uterus specimens were collected from patients with adenomyosis undergoing hysterectomy in Beijing Obstetrics and Gynecology Hospital,which included 9 proliferative endometrium and 7 secretory endometrium.EMI smooth muscle cells from the uterus were cultured and loaded with calcium ion ( Ca2 + ) fluorescent probe fluo-4/AM.The labeled cells were stimulated with the various concentration of E2 ( 1 × 102,1 × 103,1 × 104,1 × 105 pmol/L,respectively),then the changes of intracellular Ca2+ fluorescence intensity were measured by laser scanning microscopy.The most suitable concentration of E2 was selected,and the reaction difference between the EMI smooth muscle cells of two menstrual phases were also investigated; The changes of intracellular Ca2 + fluorescence intensity were detected proliferative and secretory smooth muscle cells in E2 conjugated to bovine serum albumin (17β-E2-BSA) group,cycloheximide (CHX) group,fulvestrant (ICI182780) group and pertussis toxin (PTX) group.Results ( 1 ) The cell viability of primary cultured EMI smooth muscle cells was well at 24 hours culture.(2) 1 × 102 - 1 × 105 pmol/L E2 can rapidly increase the intracellular Ca2+ fluorescence intensity within 1 min ( P < 0.01 ) ;The increased amplitudes caused by 1 × 104 pmol/L and 1 × 105 pmoL/L E2 were the most significant,but there was no significant difference between them (P >0.05).1 × 104 pmol/L was the most suitable concentration.( 3 ) With the 1 × 104 pmol/L E2,the Ca2+ fluorescence intensity changes showed no significant difference between the EMI smooth muscle cells from the proliferative phase and secretory phase uterus (P > 0.05 ).The Ca2+ fluorescence intensity changes were 646 ± 32 in 17β-E2-BSA group and 602 ±31 in CHX group,when compared with 513 ±26 and 617 ±35 in respective control group,no significant difference was observed (P > 0.05 ).The increased amplitude of 188 ± 20 in the PTX group and 302 ± 11 in ICI182780 group exhibited significant difference with 632 ± 33 and 635 ± 24 in respective control group ( P < 0.01 ).Conclusion E2 could increase the intracellular Ca2 + of EMI through a membrane receptor dependent and nongenomic mechanism of action.
4.Icariin protects against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cells differentiating into osteoblasts
Zhilie YANG ; Chenglong WANG ; Dongfeng ZHAO ; Junli CHANG ; Cheng YANG ; Yanping YANG ; Yongjun WANG
Chinese Journal of Tissue Engineering Research 2016;20(6):777-784
BACKGROUND:Osteoporosis caused by chemotherapy has become one of the serious side effects that impact the skeletal system. Icarin shows a strong anti-osteoporosis activity, which can have protective effect on osteoporosis induced by chemotherapy. OBJECTIVE: To study the protective effect and mechanism of icarin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cels differentiating into osteoblasts. METHODS:MTT assay and alkaline phosphatase (ALP) staining were used to determine the optimal protective concentration of icarin against cyclophosphamide-induced obstacle of mouse bone marrow mesenchymal stem cels differentiating into osteoblasts. mRNA expressions of osteoblast-specific transcription factors, OC, ALP, Runx2, and Wnt/β-catenin signaling pathway target genes, β-catenin, C-Myc, cyclin D1, were determined using RT-PCR method at different time after intervention with the optimal concentration of icarin. Expressions of Runx2, β-catenin, c-Myc, cyclin D1 regulated by the optimal concentration of icarin were detected using western blot assay at the protein level. RESULTS AND CONCLUSION:Cel viability and ALP activity decreased significantly in the cyclophosphamide group compared with the control group, but there was no significant difference in cel viability between icarin group and cyclophosphamide group. Icarin at 100 μmol/L showed the best protective effect against cyclophosphamide-induced obstacle of osteogenic differentiation of bone marrow mesenhymal stem cels. Compared with the control group, cyclophosphamide chemotherapy reduced the expressions of ALP, OC, Runx2 at mRNA level and Runx2 at protein level, weakened the expressions ofβ-catenin, cyclin D1 at mRNA level and active β-catenin, Cyclin D1, c-myc at protein level, and increased the expression of DKK1. Compared with the cyclophosphamide group, 100 μmol/L icarin increased the expression of osteoblast-specific transcription factors and Wnt/β-catenin signaling pathway genes at mRNA and protein levels, and reduced the expression of DKK1 protein. These results show that cyclophosphamide can lead to osteogenic differentiation disorder of mouse bone marrow mesenchymal stem cels, and in contrast, icarin shows a protective effect and its optimal intervention concentration is 100 μmol/L. Additionaly, the protective roleof icarin is probably related to activation of Wnt/β-catenin signal pathway.
5.Nail DNA and Possible Biomarkers: A Pilot Study.
Joshua PARK ; Debbie LIANG ; Jung Woo KIM ; Yongjun LUO ; Taesheng HUANG ; Soo Young KIM ; Seong Sil CHANG
Journal of Preventive Medicine and Public Health 2012;45(4):235-243
OBJECTIVES: Nail has been a substitute DNA source for genotyping. To investigate the integrity and consistency of nail DNA amplification for biomarker study, nail clippings from 12 subjects were collected at monthly intervals. The possibility of longer amplification and existence of GAPDH RNA/protein, were also investigated with three nail samples. METHODS: Three primer sets were designed for quantitative amplification of nuclear and mitochondrial genes and analysis of their consistency. The mean threshold cycles in amplification of the target genes were compared to test the consistency of polymerase chain reaction (PCR) performance among individual factors including age groups, sex, family, the nail source, and by the size of the amplification segments. RESULTS: The amplification of the target genes from nail DNA showed similar integrity and consistency between the nail sources, and among the serial collections. However, nail DNA from those in their forties showed earlier threshold cycles in amplification than those in their teens or seventies. Mitochondrial DNA (mtDNA) showed better DNA integrity and consistency in amplification of all three targets than did nuclear DNA (nucDNA). Over 9 kb of mtDNA was successfully amplified, and nested quantitative PCR showed reliable copy numbers (%) between the two loci. Reverse transcription PCR for mRNA and immunoblotting for GAPDH protein successfully reflected their corresponding amounts. Regarding the existence of RNA and protein in nails, more effective extraction and detection methods need to be set up to validate the feasibility in biomarker study. CONCLUSIONS: Nail DNA might be a feasible intra-individual monitoring biomarker. Considering integrity and consistency in target amplification, mtDNA would be a better target for biomarker research than nucDNA.
Adult
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Age Factors
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Aged
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Biological Markers/analysis
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Child
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DNA/*analysis/isolation & purification
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DNA Primers
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DNA, Mitochondrial/analysis
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Feasibility Studies
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Female
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Gene Amplification
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Humans
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Male
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Middle Aged
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Nails/*chemistry
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Pilot Projects
6.An Engineering View on Megatrends in Radiology: Digitization to Quantitative Tools of Medicine.
Namkug KIM ; Jaesoon CHOI ; Jaeyoun YI ; Seungwook CHOI ; Seyoun PARK ; Yongjun CHANG ; Joon Beom SEO
Korean Journal of Radiology 2013;14(2):139-153
Within six months of the discovery of X-ray in 1895, the technology was used to scan the interior of the human body, paving the way for many innovations in the field of medicine, including an ultrasound device in 1950, a CT scanner in 1972, and MRI in 1980. More recent decades have witnessed developments such as digital imaging using a picture archiving and communication system, computer-aided detection/diagnosis, organ-specific workstations, and molecular, functional, and quantitative imaging. One of the latest technical breakthrough in the field of radiology has been imaging genomics and robotic interventions for biopsy and theragnosis. This review provides an engineering perspective on these developments and several other megatrends in radiology.
Biological Markers/analysis
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Biomedical Engineering
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Diagnosis, Computer-Assisted/*trends
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Diagnostic Imaging/*trends
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Equipment Design
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Genomics
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Humans
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Image Processing, Computer-Assisted/*trends
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Radiology Information Systems/*trends
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Robotics
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Systems Integration
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User-Computer Interface
7.Study on HPLC Fingerprint of Miao Medicine Ardisia crenata
Xu SUN ; Chengfen YAO ; Sihong FU ; Zaipeng GONG ; Ting LIU ; Chang YANG ; Jun ZHA ; Yongjun LI
China Pharmacy 2017;28(30):4285-4288
OBJECTIVE:To establish HPLC fingerprints of Miao medicine Ardisia crenata.METHODS:HPLC method was adopted.The determination was performed on Diamonsil C18 column with mobile phase consiste of methanol-water (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was 220 nm,and column temperature was maintained at 30 ℃.The sample size was 10 μL.Using 11-O-(3',4',5'-three-o-galloylhyperin)-bergeninum as reference,HPLC fingerprints of 16 batches of samples were determined.Common identification and similarity evaluation were performed by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2012 edition).Cluster analysis of fmgerprrints was conducted.RESULTS:There were 6 common peaks in HPLC fingerprints of 16 batches of samples.The similarity among 8 batches was more than 0.9.The 16 batches of samples could be clustered into 4 categories.CONCLUSIONS:Established fingerprints can provide reference for identification and quality evaluation ofA.crenata.
8.Changes of CK,CK-MB,cTnI and electrocardiogram before and after treatment in children with pneumonia
Kaichao CHANG ; Honglin LIU ; Xia ZHANG ; Xiong WU ; Yongjun ZHANG ; Penglin HU
International Journal of Laboratory Medicine 2018;39(10):1227-1229
Objective To investigate the changes of serum creatine kinase (CK),creatine kinase isoenzyme (CK-MB),cardiac troponin I(cTnI)and electrocardiogram (ECG) before and after the treatment of pneumo-nia in children.Methods From December 2014 to December 2016,95 children with pneumonia were selected as the study group,and 48 healthy subjects who underwent the healthy assessment from December 2014 to January 2016 were selected as the control group.All children with pneumonia were treated after admission.2 mL of venous blood were collected from each research subject after the admission and patients in study group after treatment,serum was seperated,and levels of CK,CK-MB,cTnI were measured and the ECG record was conducted.Results The serum levels of CK,CK-MB and cTnI in the study group were higher than those in the control group (P<0.05);the incidences of ST segment elevation or depression,atrial premature beat,ven-tricular premature beat,sinus tachycardia and sinus bradycardia in the study group were higher than those in the control group,and the differences were statistically significant (P<0.05);the serum levels of CK,CK-MB and cTnI in the study group were lower than those before treatment,and the difference was statistically signif-icant (P<0.05);the incidences of atrial premature beat,ventricular premature beat,sinus tachycardia and si-nus bradycardia in the study group after treatment were lower than those before treatment,and the difference was statistically significant (P<0.05);the incidence of ST segment elevation or depression after treatment in the study group was lower than that before treatment,and there was no significant difference (P>0.05).Con-clusion The serum levels of CK,CK-MB,cTnI and ECG were obviously abnormal in children with pneumoni-a.After treatment,serum CK,CK-MB and cTnI levels can be reduced and ECG abnormalities can be ameliora-ted.
9.Computer-aided detection system for masses in automated whole breast ultrasonography: development and evaluation of the effectiveness.
Jeoung Hyun KIM ; Joo Hee CHA ; Namkug KIM ; Yongjun CHANG ; Myung Su KO ; Young Wook CHOI ; Hak Hee KIM
Ultrasonography 2014;33(2):105-115
PURPOSE: The aim of this study was to evaluate the performance of a proposed computer-aided detection (CAD) system in automated breast ultrasonography (ABUS). METHODS: Eighty-nine two-dimensional images (20 cysts, 42 benign lesions, and 27 malignant lesions) were obtained from 47 patients who underwent ABUS (ACUSON S2000). After boundary detection and removal, we detected mass candidates by using the proposed adjusted Otsu's threshold; the threshold was adaptive to the variations of pixel intensities in an image. Then, the detected candidates were segmented. Features of the segmented objects were extracted and used for training/testing in the classification. In our study, a support vector machine classifier was adopted. Eighteen features were used to determine whether the candidates were true lesions or not. A five-fold cross validation was repeated 20 times for the performance evaluation. The sensitivity and the false positive rate per image were calculated, and the classification accuracy was evaluated for each feature. RESULTS: In the classification step, the sensitivity of the proposed CAD system was 82.67% (SD, 0.02%). The false positive rate was 0.26 per image. In the detection/segmentation step, the sensitivities for benign and malignant mass detection were 90.47% (38/42) and 92.59% (25/27), respectively. In the five-fold cross-validation, the standard deviation of pixel intensities for the mass candidates was the most frequently selected feature, followed by the vertical position of the centroids. In the univariate analysis, each feature had 50% or higher accuracy. CONCLUSION: The proposed CAD system can be used for lesion detection in ABUS and may be useful in improving the screening efficiency.
Classification
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Humans
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Imaging, Three-Dimensional
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Mass Screening
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Radiographic Image Interpretation, Computer-Assisted
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Support Vector Machine
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Ultrasonography, Mammary*
10.Helminth Eggs Detected in Soil Samples of a Possible Toilet Structure Found at the Capital Area of Ancient Baekje Kingdom of Korea
Chang Seok OH ; Sang-Yuck SHIM ; Yongjun KIM ; Jong Ha HONG ; Jong-Yil CHAI ; Hisashi FUJITA ; Min SEO ; Dong Hoon SHIN
The Korean Journal of Parasitology 2021;59(4):393-397
Although research conducted in East Asia has uncovered parasite eggs from ancient toilets or cesspits, data accumulated to date needs to be supplemented by more archaeoparasitological studies. We examined a total of 21 soil samples from a toilet-like structure at the Hwajisan site, a Baekje-period royal villa, in present-day Korea. At least 4 species of helminth eggs, i.e., Trichuris trichiura, Ascaris lumbricoides, Clonorchis sinensis, and Trichuris sp. (or Trichuris vulpis) were detected in 3 sediment samples of the structure that was likely a toilet used by Baekje nobles. The eggs of T. trichiura were found in all 3 samples (no. 1, 4, and 5); and A. lumbricoides eggs were detected in 2 samples (no. 4 and 5). C. sinensis and T. vulpis-like eggs were found in no. 5 sample. From the findings of this study, we can suppose that the soil-transmitted helminths were prevalent in ancient Korean people, including the nobles of Baekje Kingdom during the 5th to 7th century.