1.Interferon-related gene array in predicting the efficacy of interferon therapy in chronic hepatitis B.
Jiayi WANG ; Jiajie LU ; Chen ZHOU ; Lingyao DU ; Hong TANG
Journal of Biomedical Engineering 2023;40(1):79-86
		                        		
		                        			
		                        			This study aims to clarify host factors of IFN treatment in the treatment of chronic hepatitis B (CHB) patients by screening the differentially expressed genes of IFN pathway CHB patients with different response to interferon (IFN) therapy. Three cases were randomly selected in IFN-responding CHB patients (Rs), non-responding CHB patients (NRs) and healthy participants, respectively. The human type I IFN response RT 2 profiler PCR array was used to detect the expression levels of IFN-related genes in peripheral blood monocytes (PBMCs) from healthy participants and CHB patients before and after Peg-IFN-α 2a treatment. The results showed that more differentially expressed genes appeared in Rs group than NRs group after IFN treatment. Comparing with healthy participants, IFNG, IL7R, IRF1, and IRF8 were downregulated in both Rs and NRs group before IFN treatment; CXCL10, IFIT1, and IFITM1 were upregulated in the Rs; IL13RA1 and IFI35 were upregulated in the NRs, while IFRD2, IL11RA, IL4R, IRF3, IRF4, PYHIN1, and ADAR were downregulated. The expression of IL15, IFI35 and IFI44 was downregulated by 4.09 ( t = 10.58, P < 0.001), 5.59 ( t = 3.37, P = 0.028) and 10.83 ( t = 2.8, P = 0.049) fold in the Rs group compared with the NRs group, respectively. In conclusion, IFN-response-related gene array is able to evaluate IFN treatment response by detecting IFN-related genes levels in PBMC. High expression of CXCL10, IFIT1 and IFITM1 before treatment may suggest satisfied IFN efficacy, while high expression of IL13RA1, IL15, IFI35 and IFI44 molecules and low expression of IFRD2, IL11RA, IL4R, IRF3, IRF4, PYHIN1 and ADAR molecules may be associated with poor IFN efficacy.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Healthy Volunteers
		                        			;
		                        		
		                        			Hepatitis B, Chronic/genetics*
		                        			;
		                        		
		                        			Immunotherapy
		                        			;
		                        		
		                        			Interleukin-15
		                        			;
		                        		
		                        			Leukocytes, Mononuclear
		                        			;
		                        		
		                        			Nuclear Proteins
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis/methods*
		                        			;
		                        		
		                        			Interferons/therapeutic use*
		                        			;
		                        		
		                        			Treatment Outcome
		                        			
		                        		
		                        	
2.Epigenetic regulation mechanism: roles in enamel formation and developmental defects of enamel.
Mian WAN ; Yi Ting LI ; Li Wei ZHENG ; Xue Dong ZHOU
Chinese Journal of Stomatology 2023;58(1):68-74
		                        		
		                        			
		                        			Enamel formation is a series of complex physiological processes, which are regulated by critical genes spatially and temporally. These processes involve multiple developmental stages covering ages and are prone to suffer signal interference or gene mutations, ultimately leading to developmental defects of enamel (DDE). Epigenetic modifications have important regulatory roles in gene expression during enarnel development. New technologies including high-throughput sequencing, chromatin immunoprecipitation sequencing (ChIP-seq), and DNA methylation chip are emerging in recent years, making it possible to establish genome-wide epigenetic modification profiles during developmental processes. The regulatory role of epigenetic modification with spatio-temporal pattern, such as DNA methylation, histone modification and non-coding RNA, has significantly expanded our understanding of the regulatory network of enamel formation, providing a new theoretical basis of clinical management and intervention strategy for DDE. The present review briefly describes the enamel formation process of human beings' teeth as well as rodent incisors and summarizes the dynamic characteristics of epigenetic modification during enamel formation. The functions of epigenetic modification in enamel formation and DDE are also emphatically discussed.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Epigenesis, Genetic
		                        			;
		                        		
		                        			Developmental Defects of Enamel
		                        			;
		                        		
		                        			DNA Methylation
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			Dental Enamel
		                        			
		                        		
		                        	
3.Advances in microbial transcriptomics techniques.
Yuanyuan GUO ; Yaru SUN ; Heping ZHANG
Chinese Journal of Biotechnology 2022;38(10):3606-3615
		                        		
		                        			
		                        			With the rapid development of molecular biotechnology, transcriptomics has been widely used in the study of gene expression. In recent years, the techniques for microbial transcriptomics research have also been rapidly developing. At the gene level, the way for obtaining sequence information has been developed from complementary validation of RNA fragment through DNA microarray to direct sequencing of full-length RNA. Spatially, the traditional population transcriptomics technique has been developed into spatial, single cell and epigenetic transcriptomics studies. With the application of transcriptomics techniques in the field of microbial research, the corresponding defects were gradually revealed and constantly improved. In this paper, the traditional and new transcriptomics techniques in the field of microbial research are summarized to provide reference for microbial transcriptomics research.
		                        		
		                        		
		                        		
		                        			Transcriptome
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			Sequence Analysis
		                        			;
		                        		
		                        			RNA
		                        			;
		                        		
		                        			Biotechnology
		                        			
		                        		
		                        	
4.Visualization analysis of microfluidics research status.
Wei WEI ; Ruijun WU ; Xiaodong SANG ; Tianyu LIANG ; Zhifei LI ; Zhi LI ; Yang YANG ; Yue SU
Journal of Biomedical Engineering 2022;39(3):551-560
		                        		
		                        			
		                        			Microfluidics is the science and technology to manipulate small amounts of fluids in micro/nano-scale space. Multiple modules could be integrated into microfluidic device, and due to its advantages of microminiaturization and controllability, microfluidics has drawn extensive attention since its birth. In this paper, the literature data related to microfluidics research from January 1, 2006 to December 31, 2021 were obtained from Web of Science Core Collection database. CiteSpace 5.8.R3 software was used for bibliometrics analysis, so as to explore the research progress and development trends of microfluidics research at home and abroad. Based on the analysis of 50 129 articles, it could be seen that microfluidics was a hot topic of global concern, and the United States had a certain degree of authority in this field. Massachusetts Institute of Technology and Harvard University not only had a high number of publications, but also had strong influence and extensive cooperation network. Combined with ultrasonic, surface modification and sensor technology, researchers constructed paper-based microfluidic, droplet microfluidic and digital microfluidic platforms, which were applied in the field of immediate diagnosis, nucleic acid and circulating tumor cell analysis of in vitro diagnosis and organ-on-a-chip. China was one of the countries with a high level of research in the field of microfluidics, while the industrialization of high-end products needed to be improved. As people's demand for disease risk prediction and health management increased, promoting microfluidic technological innovation and achievement transformation is of great significance to safeguard people's life and health.
		                        		
		                        		
		                        		
		                        			China
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Microfluidic Analytical Techniques
		                        			;
		                        		
		                        			Microfluidics
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			
		                        		
		                        	
5.Application of chromosomal microarray analysis in prenatal diagnosis of pregnant women with advanced age.
Shuting YANG ; Yali ZHAO ; Xinxin TANG ; Zhiwei WANG ; Dengping LIU ; Jinglu ZHANG ; Ying GU ; Leilei WANG
Chinese Journal of Medical Genetics 2021;38(2):101-107
		                        		
		                        			OBJECTIVE:
		                        			To assess the value of chromosomal microarray analysis (CMA) for the detection of fetal anomalies among pregnant women with advanced age.
		                        		
		                        			METHODS:
		                        			CMA results of 562 cases, in addition with the outcome of pregnancy and neonatal follow-up were reviewed.
		                        		
		                        			RESULTS:
		                        			Among the 562 amniotic fluid samples, 73 cases (12.99%) of fetal chromosomal abnormalities were detected, which included 21 cases (3.73%) of chromosomal aneuploidies and 52 cases (9.25%) of copy number variations (CNVs). The latters included 27 cases of pathological CNVs (4.80%), 4 cases of possible pathogenic CNVs (0.71%) and 42 cases of variants with unknown clinical significance (7.47%). Compared with those under 35, the detection rate of fetal chromosomal aneuploidies for women with advanced age was higher under the indications of voluntary test, abnormal ultrasonic structures, abnormal ultrasonic soft index and risks indicated by non-invasive prenatal testing (NIPT). No significant difference was found in the detection rate of CNVs between those ≥35 and <35 and between those with age factor only and with additional indications (P> 0.05). 552 cases (98.22%) of pregnant women have completed the followed up. Among 31 women with pathological and possible pathogenic fetal CNVs detected by CMA, 25 had terminated the pregnancy, 6 (19.35%) have delivered without obvious abnormality. 41 pregnant women with fetal CNVs of unknown clinical significance have completed the follow up, among whom 3 had terminated the pregnancy, 1 newborn was found with malformation after birth, which yielded an abnormal pregnancy rate of 9.76%. 480 pregnant women with negative CMA results have completed the follow up, among whom 5 (1.04%) had abnormal pregnancy or delivered a child with birth defect.
		                        		
		                        			CONCLUSION
		                        			There is a certain difference between the outcome of pregnancy predicted by CMA testing and the actual outcome. The pregnancies with fetal CNVs with unknown clinical significance detected by CMA have a high adverse rate, which should attract clinical attention. CMA testing should be recommended for pregnant women with advanced age regardless of whether they have other symptoms. CMA combined with other detection methods is the trend for prenatal diagnosis.
		                        		
		                        		
		                        		
		                        			Aneuploidy
		                        			;
		                        		
		                        			Chromosome Aberrations
		                        			;
		                        		
		                        			DNA Copy Number Variations
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Maternal Age
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			Pregnancy
		                        			;
		                        		
		                        			Prenatal Diagnosis
		                        			
		                        		
		                        	
6.Methodological Evaluation of Microarray in the Detection of α-Thalassemia.
Peng-Fei CAI ; Liu-Qun QIN ; Shi-Qiang LUO ; Li-Zhu CHEN ; Qing-Yan ZHONG ; Jing-Ren WANG ; Qiu-Hua WANG ; Jun HUANG ; Ti-Zhen YAN
Journal of Experimental Hematology 2021;29(6):1907-1910
		                        		
		                        			OBJECTIVE:
		                        			To proceed the clinical evaluation of DNA microarray for thalassemia gene detection.
		                        		
		                        			METHODS:
		                        			Peripheral blood samples of 166 thalassemia gene test subjects were collected and tested for thalassemia genes by microarray chip method and Gap-PCR method combined with PCR-reverse dot blot hybridization method according to double-blind control test. The specificity, sensitivity, positive predictive value, negative predictive value, and total coincidence rate of the microarray chip method were evaluated. When the two methods were inconsistent, multiplex ligation dependent probe amplification (MLPA) was used to verify the deletional α-thalassemia.
		                        		
		                        			RESULTS:
		                        			Compared with Gap-PCR method, specificity, sensitivity, positive predictive value, negative predictive value, Youden index, and total coincidence rate of microarray chip method was 100% (70/70), 96.88% (93/96), 100% (93/93), 95.89% (70/73), 0.969, and 97.59% (162/166), respectively, while compared with PCR-reverse dot blot hybridization method was 100% (125/125), 100% (41/41), 100% (41/41), 100% (125/125), 1, and 100% (166/166), respectively.
		                        		
		                        			CONCLUSION
		                        			The microarray chip method for α-thalassemia gene detection shows the advantages of high specificity, sensitivity, and throughput.
		                        		
		                        		
		                        		
		                        			Genetic Testing
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Multiplex Polymerase Chain Reaction
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			alpha-Thalassemia/genetics*
		                        			
		                        		
		                        	
7.Application of DNA Microarray in Genetic Mutation Detection in Patients with Thalassemia.
Liu-Qun QIN ; Ti-Zhen YAN ; Shi-Qiang LUO ; Peng-Fei CAI ; Li-Zhu CHEN ; Qing-Yan ZHONG ; Jing-Ren WANG ; Qiu-Hua WANG ; De-Jian YUAN ; Jun HUANG
Journal of Experimental Hematology 2021;29(5):1561-1565
		                        		
		                        			OBJECTIVE:
		                        			To perform dried blood spots thalassemia gene detection in patients with positive blood phenotypes by microarray technology, and evaluate its value in clinical detection.
		                        		
		                        			METHODS:
		                        			DNA samples were extracted from dried blood spots of 410 patients. Microarray technology was used to detect 3 deletion and 3 non-deletion types of α-thalassemia and 19 β-thalassemia point mutations which were common gene mutions in China.
		                        		
		                        			RESULTS:
		                        			There were 357 positive cases in all the 410 tested samples with the positive rate 87.07%, among which 299 cases (72.93%) carried deletion or point mutations of α-thalassemia, 29 cases (7.07%) carried point mutations of β-thalassemia and 29 cases (7.07%) carried gene mutations of complex αβ-thalassemia syndrome. The mutations of α-thalassemia were involved with --
		                        		
		                        			CONCLUSION
		                        			The most common genetic mutations are --
		                        		
		                        		
		                        		
		                        			China
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Mutation
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			alpha-Thalassemia/genetics*
		                        			;
		                        		
		                        			beta-Thalassemia/genetics*
		                        			
		                        		
		                        	
8.Analysis of the Differential Expression of circRNA in Acute Myeloid Leukemia by GEO Database.
Wei QIN ; Si-Xuan QIAN ; Xiao-Hui CAI ; Xu-Zhang LU ; Hong-Ying CHAO
Journal of Experimental Hematology 2021;29(6):1719-1726
		                        		
		                        			OBJECTIVE:
		                        			To investigate the difference expression of circular RNA (circRNA) in acute myeloid leukemia (AML) by using bioinformatics method.
		                        		
		                        			METHODS:
		                        			The microarray chip data of AML was searched and downloaded from the Gene Expression Omnibus (GEO) of the National Center for Bioinformatics (NCBI). The differences between AML samples and control samples were analyzed by R software. The interaction between deregulated circRNA, miRNA and mRNA were predicted by miranda software and miRTarBase software. The circRNA-miRNA-mRNA regulatory network was constructed by using the cytoHubba plugin based on the Cytoscape software.
		                        		
		                        			RESULTS:
		                        			A total of 203 differential expression of circRNAs were finally collected, including down-regulated 161 circRNAs and up-regulated 42 circRNAs. CircRNA/miRNA/mRNA interaction network was constructed through software prediction. hsa_circ_0001080, hsa_circ_0004511, hsa_circ_0054211, hsa_circ_0001944 may be positively regulated the gene expression in AML.
		                        		
		                        			CONCLUSION
		                        			Abnormal expression of circRNA in AML may become a new target for AML treatment.
		                        		
		                        		
		                        		
		                        			Gene Expression
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Leukemia, Myeloid, Acute/genetics*
		                        			;
		                        		
		                        			MicroRNAs/genetics*
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			RNA, Circular
		                        			
		                        		
		                        	
9.Development of a novel high throughput brain-on-chip with 3D structure and its application in evaluation of pesticide-induced-neurotoxicity.
Chenyu ZHAO ; Haidi LI ; Xiaoping CHEN
Chinese Journal of Biotechnology 2021;37(7):2543-2553
		                        		
		                        			
		                        			We designed and fabricated a novel high throughput brain-on-chip with three dimensional structure with the aim to simulate the in vivo three-dimensional growth environment for brain tissues. The chip consists of a porous filter and 3D brain cell particles, and is loaded into a conventional 96-well plate for use. The filter and the particle molds were fabricated by using computer modeling, 3D printing of positive mold and agarose-PDMS double reversal mold. The 3D cell particles were made by pouring and solidifying a suspension of mouse embryonic brain cells with sodium alginate into a cell particle mold, and then cutting the resulting hydrogel into pieces. The loaded brain-on-chip was used to determine the neurotoxicity of pesticides. The cell particles were exposed to 0, 10, 30, 50, 100 and 200 µmol/L of chlorpyrifos or imidacloprid, separated conveniently from the medium by removing the porous filter after cultivation. Subsequently, cell proliferation, acetylcholinesterase activity and lactate dehydrogenase release were determined for toxicity evaluation. The embryonic brain cells were able to grow and proliferate normally in the hydrogel particles loaded into the filter in a 96-well plate. Pesticide neurotoxicity test showed that both chlorpyrifos and imidacloprid presented dose-dependent inhibition on cell growth and proliferation. Moreover, the pesticides showed inhibition on acetylcholinesterase activity and increase release of lactate dehydrogenase. However, the effect of imidacloprid was significantly weaker than that of chlorpyrifos. In conclusion, a novel brain-on-chip was developed in this study, which can be used to efficiently assess the drug neurotoxicity, pharmacodynamics, and disease mechanism by combining with a microtiterplate reader.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Brain
		                        			;
		                        		
		                        			Chlorpyrifos/toxicity*
		                        			;
		                        		
		                        			Culture Media
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			Pesticides/toxicity*
		                        			
		                        		
		                        	
10.Identification and validation of differential expression of miR-455-5p in plasma of children with Kawasaki disease.
Jie JIANG ; Zhuoying LI ; Xin LI ; Shentang LI ; Zuocheng YANG
Journal of Central South University(Medical Sciences) 2020;45(6):673-677
		                        		
		                        			OBJECTIVES:
		                        			To provide clues for further study of the relationship between miRNAs and Kawasaki disease (KD) development, and to provide molecular markers for ultimately improve the rate of early diagnosis for KD.
		                        		
		                        			METHODS:
		                        			We collected acute, recovery KD children's plasma and normal samples, then used the miRNAs Assay Chip to screen the differentially expressed miRNAs in the plasma from KD children. Subsequently, miR-455-5p, which had identified via miRNAs assay chip, was validated by quantitative real-time PCR via independent cohort.
		                        		
		                        			RESULTS:
		                        			According to the results of miRNAs Assay chip, we identified a miRNAs panel including 5 miRNAs significantly up-regulated and 5 miRNAs remarkably down-regulated in the plasma from KD children compared to the normal control; miR-455-5p in both of acute and recovery KD children's plasma was remarkably lower than that in the normal control (<0.001, =0.013, respectively), and miR-455-5p was also significantly lower than that in the recovery of KD children (=0.007) by independent cohort validation.
		                        		
		                        			CONCLUSIONS
		                        			There are significantly differentially expressed circulating miRNAs between the KD children and normal control. We identified 10 miRNAs dysregulation in the KD children's plasma compared with the normal group. Circulating miR-455-5p in both of acute and recovery KD children's plasma is remarkably lower than that in the normal control, and miR-455-5p may considered as a marker to show the recovery process of KD children. Plasma specific circulating miRNAs play an important role in the early diagnosis of KD and become the new molecular marker of KD in the future.
		                        		
		                        		
		                        		
		                        			Biomarkers
		                        			;
		                        		
		                        			Child
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			MicroRNAs
		                        			;
		                        		
		                        			genetics
		                        			;
		                        		
		                        			Mucocutaneous Lymph Node Syndrome
		                        			;
		                        		
		                        			genetics
		                        			;
		                        		
		                        			Oligonucleotide Array Sequence Analysis
		                        			;
		                        		
		                        			Real-Time Polymerase Chain Reaction
		                        			
		                        		
		                        	
            
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