1.Clinical Application of Tidal Flow-volume Measurement and Its Impact Factors
Jingjuan GUO ; Zhiwei YU ; Juanjuan XIE
Journal of Medical Research 2009;38(8):55-57
Objective To study the clinical application and its impact factors of tidal flow - volume measurement in infants with bronchiolitis and pneumonia. Methods Thirty infants with bronchiolitis and thirty infants with pneumonia were studied. Tidal breathing flow- volume loops were obtained after the patients were sedated. The tidal volume per kilogram(VT/kg) ,the fraction of tidal volume ex-heled at peak tidal expiratory flow (PTEF) to total tidal volume (VPTEF/VE), and the fraction of exhaled time at PTEF to total expirato-ry time (TPTEF/TE) were measured. Results VT/kg was not significant different between the bronehiolitis group and the pneumonia group. VPTEF/VE and TPTEF/TE were lower in the bronehiolitis group than those in the pneumonia group. Conclusion The measure-ment of tidal breathing flow - volume loops is a good technology, which would be helpful to evaluate airway obstruction in wheezing in-fants. The impact factors should be controlled during the measurement. Otherwise, the results would be affected.
2.Association of uromodulin gene polymorphism with diabetic kidney disease in Han population in Tianjin of China
Jun GUO ; Liming CHEN ; Baocheng CHANG ; Miaoyan ZHENG ; Juanjuan WEN
Chinese Journal of Nephrology 2016;32(5):339-344
Objective To investigate the association of single nucleotide polymorphism (SNP) rs13333226 in uromodulin (UMOD) gene with diabetic kidney diseases (DKD) in Han population in Tianjin,China.Methods A total of 210 type 2 diabetes (T2DM),90 normal controls (NC) and 280 DKD patients were recruited.According to the level of estimated glomerular filtration rate (eGFR),the DKD subjects were further subdivided into three groups:GFR≥90 ml/min group (n=105),60 ml/mim≤GFR < 90 ml/min group (n=84) and GFR < 60 ml/min group (n=91).Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for UMOD rs13333226C genotyping.Results The frequencies of AA,GA,GG genotype were 27.8%,58.9%,13.3% in NC group and 41.0%,48.6%,10.5% in T2DM group and 54.3%,36.1%,9.6% in DKD group.The frequency of G allele was 42.8% in NC group,34.8% in T2DM group and 27.7% in DKD group.The genotype distribution of UMOD was statistically significant between NC group and DKD group,and between T2DM group and DKD group (P < 0.05).G allele of UMOD was an independent protective gene polymorphism of DKD in Logistic regression (B=-0.248,Wald=8.012,P=0.021,OR=0.780,95% CI 0.612-0.968).Conclusion The G allele of UMOD gene may be an independent protective factor of DKD in Han population in Tianjin,China.
3.Evaluation of Antioxidant Activity of Flaxseed Extracts by 2, 2-Diphenyl-1-picrylhydrazyl-High Performance Liquid Chromatography Assay
Mingyu LIU ; Min LI ; Juanjuan CHEN ; Xian GUO ; Xiaojun YAN
Chinese Journal of Analytical Chemistry 2015;(2):245-250
The free radical scavenging effect of flaxseed was screened by HPLC-DPPH ( 2 , 2-diphenyl-1-picrylhydrazyl-high performance liquid chromatography assay ) and colorimetric DPPH methods. To test the effectiveness of the approach, three Lignans ( secoisolariciresinol diglucoside ( SDG ) , secoisolariciresinol ( SECO) and enterodiol( ED) ) with antioxidative properties were investigated both in monomer and mixture. HPLC conditions were optimized using following methods: Waters XBridge C18 was used as stationary phase, acetonil/H2 O was used as mobile phase and detective wavelength was set at 280 nm. Antioxidant activity of standards was investigated by reaction with or without DPPH radical for 20 min as sample and control, respectively. Both of them were analyzed by high performance liquid chromatography. According to the changes of amount of sample and control, the antioxidant activities of standards were calculated as following order:SDG>SECO>ED. Based on above DPPH-HPLC assay and UPLC-Q-TOF-MS, antioxidants extracted from flaxseed were separated, identified and screened. The radical scavenging activities were in the following order:SDG isomer (5)>SDG (4)>7α-[(β-D-glupyranosyl) oxy]-1-methoxyisolariciresinol (1)>(6R,7R, 8S)-1-methoxyisolariciresinol (2)>herbacetindiglucoside (3). It indicated that the HPLC-DPPH assay could be successfully used for the antioxidant activity screening of complex flaxseed extract.
4.Research on miR-497 inhibiting the expression of inflammation-related genes in colon cancer cell line HCT116 using gene expression microarray
Juanjuan WANG ; Ruihong YANG ; Fuhua WANG ; Xiangyun GUO ; Xiaoyu LI ; Shufeng LIANG ; Sutang GUO
Cancer Research and Clinic 2017;29(1):1-6
Objective To analyze the effect of miR-497 high expression on the gene expression profile of colon cancer cell line HCT116. Methods MiR-497 high expressing colon cancer cell model HCT116-497 and negative control HCT116-CON were established by lentiviral transduction. The human (V2) gene expression microarray was used to identify genes that were differentially expressed between colon cancer cells overexpressing miR-497 and the controls. The candidates were subjected to the gene ontology (GO) and KEGG pathway enrichment analysis by Molecule Annotation System 3.0 (MAS3.0). The differential expression of representative genes relative to inflammation were confirmed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results Of all the differently expressed genes, 582 genes were down-regulated by at least 3-folds, which were enriched in inflammation-related signaling pathways in colon cancer cells overexpressing miR-497. The decrease in 15 representative genes was validated by qPCR. Compared with those in HCT116-CON cells, expressions of 10 genes in HCT116-497 cells, including CACNB1, FOS, IL-29, RPS6KA2, TNFSF15, IL-11, INHBC, CSF1R, JAK3 and IL-2Rβ, were decreased significantly, and there were statistical differences (all P< 0.05) Conclusion MiR-497 inhibits the mRNA expression of inflammation-related genes in colon cancer cell line HCT116.
5.Genetic Polymorphism of Nine Non-CODIS STR Loci in Hu-nan Province-based Chinese Han Population
Juanjuan GUO ; Ying LIU ; Yadong GUO ; Jie YAN ; Yunfeng CHANG ; Jifeng CAI ; Ting LU ; Lagabaiyila ZHA
Journal of Forensic Medicine 2014;(6):441-445
Objective To determine the allelic frequency distribution and genetic parameters of nine non-CODIS DNA index systems of the short tandemrepeat (STR ) loci (D2S1772, D6S1043, D7S3048, D8S1132, D11S2368, D12S391, D13S325, D18S1364, and GATA198B05). Methods A total of 353 blood samples were collected, extracted, amplified, and analyzed fromunrelated healthy individuals of Han na-tionality in Hunan Province, China. Results O ne hundred and fourteen alleles were observed in the pop-ulation with corresponding allelic frequencies ranged from0.001 0 to 0.323 0. For all the nine non-CODIS STR loci, the observed genotypic data showed no significant deviations fromthe Hardy-W einberg equi-librium. The Ho, He, PIC, D P, and PE of the studied non-CODIS STR loci ranged from0.108 0 to 0.195 0, 0.805 0 to 0.892 0, 0.770 0 to 0.860 0, 0.925 0 to 0.966 0 and 0.607 0 to 0.780 0, respectively. Conclusion N ine non-CODIS STR loci have high degrees of polymorphisms, which may be useful in in-dividual forensic identification and parentage testing in forensic practice.
6.Protective effect of Huaxia shallot preparation on human umbilical vein endothelial cell injury induced by oxidized low density lipoprotein and its mechanism
Jiemei ZHANG ; Jie GUO ; Xin TU ; Zhaohong SHI ; Jianjun HAO ; Yuhe KE ; Jiangfeng GUAN ; Juanjuan HE
Journal of Integrative Medicine 2007;5(6):675-80
OBJECTIVE: To observe the protective effect of Huaxia shallot preparation on human umbilical vein endothelial cell (HUVEC) injury induced by oxidized low density lipoprotein (Ox-LDL) in vitro. METHODS: Ox-LDL was prepared and identified, and HUVECs were cultured. After 2-hour intervention of different drugs and 24-hour following intervention of Ox-LDL, the number of HUVECs was observed by phase contrast optical microscope and the activity of the HUVECs was observed by methyl thiazolyl tetrazolium (MTT) technique. Superoxide dismutase (SOD) activity and nitric oxide (NO) content were assayed by respective kit. The protein expressions and mRNA levels of peroxisome proliferators activated receptor gamma(PPAR-gamma) and endothelial nitric oxide synthase (eNOS) were measured by western blot technique and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Ox-LDL could increase the apoptosis rate of the HUVECs and decrease the NO release as compared with the blank control group (P<0.05). These effects induced by Ox-LDL were all significantly inhibited by Huaxia shallot preparation. It could up-regulate the protein expressions and mRNA levels of PPAR-gamma and eNOS significantly (P<0.05). Huaxia shallot preparation could decrease the apoptosis rate of the HUVECs. CONCLUSION: Ox-LDL may be involved in the initiation and progression of atherosclerosis by injuring the endothelial cells directly and may cause the endothelial dysfunction. Huaxia shallot preparation can protect against Ox-LDL induced endothelial cell injury by up-regulating the protein expressions and mRNA levels of PPAR-gamma and eNOS. It suggests that Huaxia shallot preparation may play a role in the prevention and treatment of cardiovascular disease.
7.Significance and expression of miR-7 in activated CD4+T cells
Hualin XU ; Juanjuan ZHAO ; Panpan CUI ; Mengmeng GUO ; Yijing TAO ; Chao CHEN ; Lin XU
Chinese Journal of Immunology 2016;32(10):1419-1423
Objective:To investigate the change of expression of miR-7 in activated CD4+ T cells in vitro, and preliminary explore its possible significance. Methods: CD4+CD62L+T cells was purified from splenocytes of FVB mice by magnetic cell sorting system (MACS). After stimulation with anti-CD3/CD28 antibody,the relative expression of miR-7 was examined by Real-time PCR, and the expression level of CD69 molecular was analyzed by FACS. Furthermore,the relative expression of miR-7 in CD4+T cells was detected at different time points during stimulation. With the treatment of ERK inhibitor PD98059,change of miR-7 expression was de-termined by Real-time PCR. Meanwhile, the proliferation of CD4+T cells was examined by CCK-8 assay and the expression level of CD69 and CD62L molecular were analyzed by FACS. Finally,the expression of cytokines IL-6,IL-10,and IFN-γ were determined by Real-time PCR. Results:Compared with control group,the relative expression of miR-7 was increased significantly after stimulation with anti-CD3/CD28 antibody,as well as expression level of CD69 molecular was augmented(P<0. 05). In contrasted with 0 h and 24 h,the expression of miR-7 was significantly increased after 48 h and 72 h during stimulation(P<0. 05). Furthermore,the relative expression of miR-7 was significantly declined in CD4+T cells in ERK inhibitor PD98059 treatment group. Finally, the expression level of CD69 molecular,as well as cytokines IL-6, IL-10 and IFN-γ, were also decreased significantly ( P<0. 05 ) . Conclusion: The relative expression of miR-7 was significantly increased in activated CD4+T cells,closely related to ERK pathway,which provided an important foundation for successive research work on exploring the functional role of miR-7 in the CD4+T cells.
8.Influence of microRNA-7 knock down on development of CD4+SP cells in murin-ethymus
Yijing TAO ; Shunfei ZHU ; Chao CHEN ; Juanjuan ZHAO ; Mengmeng GUO ; Yixiong ZHANG ; Nalin QIN ; Lin XU
Chinese Journal of Immunology 2015;(9):1173-1177
Objective:To investigate the influence of miR-7 knock down on the development of CD 4+SP cells in the thymus in mice,and preliminary explore its possible mechanism.Methods:The changes of volume ,weight and total cell counts of thymus in miR-7 knock down (miR-7KD) mice were observed compared with Wild-type(WT)mice;the pathological changes of thymus were observed by HE staining.FACS analysis was performed on the proportion ,as well as the expression level of CD44 and CD62L,of thymus CD4+single positive (SP) cells.Meanwhile,the proliferation percentage of CD4+SP cells was measured by Ki-67 staining.The apoptosis percentage of CD4+SP cells was analyzed by FACS.The changes on the transduction of ERK 1/2 pathways were determined by Western blot.Results:Compared with WT mice ,the size,weight and total cell number of thymus were marked reduced in miR-7KD mice( P<0.05 );moreover ,pathological change also was presented.The proportion and total cell number of thymus CD 4+SP cells were marked decreased ( P<0.05 ).Furthermore ,the expression level of CD 44 and proliferation percentage ,as well as apoptosis percentage ,of CD4+SP cells were obviously increased (P<0.05),however,the expression level of CD62L of CD4+SP cells were decreased (P<0.05). Finally,the level of total ERK1/2 and phosphor-ERK1/2 was decreased obviously ( P<0.05 ).Conclusion: miR-7 knock down can affect the development of CD 4+SP cells in the thymus , which might be closely related to the cell activation state and altered the transduction of ERK1/2 pathways.
9.Genetic Polymorphisms of 21 STR Loci in Hunan Province-based Han Population
Ying ZOU ; Juanjuan GUO ; Qingpeng LI ; Daohong ZUO ; Jinshan LIU ; Yadong GUO ; Jie YAN ; Lagabaiyila ZHA ; Jifeng CAI ; Lingmei LAN
Journal of Forensic Medicine 2016;32(5):356-362
ObjectiveTo investigate the genetic polymorphisms of 21 short tandem repeat(STR)loci (D3S1358, D13S317, D7S820, D16S539, Penta E, D2S441, TPOX, TH01, D2S1338, CSF1PO, Penta D, D10S1248, D19S433, vWA, D21S11, D18S51, D6S1043, D8S1179, D5S818, D12S391 and FGA). Methods A total of 560 blood samples were collected from unrelated healthy individuals of Han population in Hunan Province. Chelex-100 extraction method was applied to the extraction of genomic DNA, and an AGCU EX22 Kit and 9700 STR amplification was used in amplification reactions. The products were separated and analyzed on 310 Genetic Analyzer.ResultsA total of 248 alleles were observed, the al-lelic frequencies ranging from 0.001 to 0.518. Observation of genotype distributions for each locus showed no deviations from Hardy-Weinberg equilibrium exceptPentaE(P=0.023). The combined pow-er of discrimination, combined power of exclusion, and combined matching probability of the 21 STR loci were approximately 0.999 999 999 999 999 999 999 999 8, 0.999 999 998, and 1.36×10-25, respectively. ConclusionThe 21 STR loci show high polymorphisms in the Han population, which can provide valu-able data and a theoretical basis for forensic individual identification and paternity testing.
10.Effect of microRNA-7 knockdown on pathology of Enterotoxin-induced murine acute lung injury
Juanjuan ZHAO ; Hualin XU ; Mengmeng GUO ; Yijing TAO ; Ya ZHOU ; Chao CHEN ; Nalin QIN ; Jing ZHENG ; Dan TIAN ; Lin XU
Chinese Journal of Immunology 2016;32(9):1257-1261
Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.