2.Preparation and performance assessment of Gamma-peptide nucleic acid gene chip detection system based on surface plasmon resonance.
Qingye OU ; Dayong GU ; Niqi ZHANG ; Jian'an HE ; Yonghong SHAO ; Lei SHI ; Chunxiao LIU ; Chunzhong ZHAO ; Yunqing XU
Journal of Biomedical Engineering 2013;30(6):1326-1329
The aim of this study was to build a gene chip system with surface plasmon resonance (SPR) technique, for which Gamma-peptide nucleic acid (Gamma-PNA) functioned as a probe, in order to improve sensitivity and its specificity. With the use of self-assembled monolayer (SAM) technology, surface chemistry of two-dimensional structure was used. Gamma-PNA was designed according to the bioinformatics, and was plated on the SPR chip modified by SAM. Subsequently, relevant parameters of the experiment were ensured and optimized. The results showed that the performances of Gamma-PNA probe was little affected by the ion concentration of buffer, and it had a strong light signal in a stable state. As the ion concentration was 0, there were still good hybrid reactions; pH value had less influence upon Gamma-PNA probe, and acid environment of buffer could be better. Gamma-PNA probe combined with sensor technologies achieved made the probe with dispensable labels and real-time detection. It also improved the efficiency of the hybridization and the stability, providing the foundation for clinical application.
Nucleic Acid Hybridization
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Nucleic Acid Probes
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Oligonucleotide Array Sequence Analysis
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methods
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Peptide Nucleic Acids
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genetics
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Sensitivity and Specificity
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Surface Plasmon Resonance
3.Type IV secretion system in Helicobacter pylori: a new insight into pathogenicity.
Qiao ZHONG ; Shi-he SHAO ; Lei-lei CUI ; Run-hong MU ; Xiao-li JU ; Su-rong DONG
Chinese Medical Journal 2007;120(23):2138-2142
OBJECTIVETo review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.
DATA SOURCESThe data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.
STUDY SELECTIONMainly original articles and critical reviews written by major pioneer investigators of this field were selected.
RESULTSThe research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.
CONCLUSIONST4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.
Bacterial Proteins ; metabolism ; DNA-Binding Proteins ; Gene Transfer, Horizontal ; Helicobacter pylori ; genetics ; metabolism ; pathogenicity ; Multigene Family
4.Comparative study of direct digital radiography and film-screen radiography in diagnosis of asbestosis.
Jian-hua SU ; Ling MAO ; He-ping XIAO ; Qin SUN ; Jin SHI ; Shao-wei ZHOU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2012;30(11):825-828
OBJECTIVETo evaluate the feasibility of direct digital radiography (DDR) in the diagnosis of asbestosis, and to analyze the difference and similarity between DDR and film-screen radiography (FSR) in terms of the radiographic features of asbestosis.
METHODSA total of 60 cases of asbestosis underwent FSR and DDR of the chest in the same day. The FSR and DDR findings were compared with respect to shapes and profusion of small opacities, pleural abnormality, and diagnostic stages.
RESULTSThe patients showed "s", "t", and "p" small opacities on chest images, with irregular "s" and "t" ones predominating (FSR: 95.0%; DDR: 91.7%). The small opacities were widely distributed in six lung zones, especially in middle and lower zones. The shapes and distribution of small opacities did not differ significantly between FSR and DDR findings (P > 0.05). For all the 60 cases, the two radiographies demonstrated a concordance rate of 64.2% (231/360) for the profusion of small opacities in lung zones (κ = 0.62, 95%CI: 0.54 ∼ 0.69), and for the 43 cases (258 lung zones) who displayed identical small opacity shapes on the two radiographies, the concordance rate was 81.0% (209/258) (κ = 0.79, 95%CI: 0.72 ∼ 0.87). FSR revealed 10 cases (16.7%) of pleural thickening, compared to 12 cases (20.0%) on DDR (P > 0.05). FSR revealed 53 cases (88.3%) of stage I asbestosis and 7 cases (11.7%) of stage II asbestosis, compared to 51 cases (85.0%) and 9 cases (15.0%) on DDR (P > 0.05). There was no significant difference in diagnostic stages between the two radiographies (P > 0.05), demonstrating a concordance rate of 93.3% (56/60) (κ = 0.71, 95%CI: 0.45 ∼ 0.98).
CONCLUSIONDDR is similar to FSR in determining the shapes, distribution, and profusion of small opacities, pleural abnormality, and diagnostic stages.
Aged ; Aged, 80 and over ; Asbestosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Radiographic Image Enhancement ; Radiography, Thoracic ; methods
5.Karyotypic and IPSS grouping of primary myelodysplastic syndromes patients: a comparison between FAB- and WHO-classification.
Ming-hua YU ; Shi-he LIU ; Ying-qi SHAO ; Yu-shu HAO ; Zhi-jian XIAO
Chinese Journal of Hematology 2004;25(8):482-485
OBJECTIVETo compare the results of cytogenetic and IPSS grouping of primary myelodysplastic syndromes (pMDS) patients classified by FAB- or WHO classification.
METHODSTwo hundred and thirty seven cases of pMDS who were previously classified according to FAB criteria were reclassified with WHO classification. A comparison was made between the results of the two classifications.
RESULTSFor the detection rates of cytogenetic abnormality and its risks group, there was no difference among the FAB subgroups but the detection rate was different between the WHO refractory cytopenia with multilineage dysplasia (RCMD) and RA subgroups (74.4% and 42.5%, respectively) (P < 0.001). The percentage of good karyotype abnormalities in RA (65.0%) was higher than that in RCMD (24.4%) (P < 0.001), and the percentages of intermediate and poor karyotype abnormalities in RCMD (48.9% and 26.7%, respectively) were higher than that in RA (27.5% and 7.5%, respectively) (P < 0.05). There was a good correlation between the subgroups and IPSS risk groups for both the WHO classification and the FAB classification, but the WHO classification further reflected the differences between RCMD and RA and RAEB-I and RAEB-II subgroups. The percentage of low-risk group in RCMD (1.1%) was lower than that in RA (10.0%) (P < 0.05), and the percentage of high-risk group in RAEB-II (30.5%) was higher than that in RAEB-I(0) (P < 0.001).
CONCLUSIONFor the correlation between subgroups and cytogenetic abnormalities and IPSS risk groups, the WHO-classification is better than the FAB-classification.
Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; classification ; genetics ; pathology ; Prognosis ; Severity of Illness Index ; World Health Organization
6.Community-based survey on human immunodeficiency virus infection among injection drug users in Sichuan, China.
Yu-Hua RUAN ; Yi-Xin HE ; Lian-Zhi XIE ; Kang-Lin CHEN ; Shi-Zhu LIU ; Feng ZHOU ; Guang-Ming QIN ; Yi-Ming SHAO
Chinese Journal of Preventive Medicine 2004;38(5):296-299
OBJECTIVETo investigate prevalence of human immunodeficiency virus (HIV) infection and risk factors for its transmission among injection drug users (IDUs) in Liangshan Yizu Autonomous Prefecture of Sichuan Province, China.
METHODSA community-based survey was conducted to investigate demographic characteristics, pattern and frequency of sharing injection equipment, and sexual behaviors in IDUs. Blood samples were also collected from them to detect for antibodies against HIV and syphilis.
RESULTSA total of 379 subjects were recruited with informed consent for study through community outreach and peer recruiting methods. Their prevalence of HIV infection was 11.3% (43/379). Ethnicity, frequency of sharing syringes and cotton swab during the past three months and syphilis infection associated with HIV infection by univariate analysis using chi-square test. Multivariate logistic regression analysis showed odds ratio of frequency of sharing syringes for HIV infection during the past three months was 2.28 (95% CI 1.18 - 4.43), and that for syphilis infection 3.10 (95% CI 1.48 - 6.48).
CONCLUSIONFrequency of sharing syringes during the past three months associated with syphilis and HIV infection.
Adolescent ; Adult ; China ; epidemiology ; Disease Transmission, Infectious ; Female ; HIV Infections ; epidemiology ; transmission ; Humans ; Logistic Models ; Male ; Needle Sharing ; adverse effects ; Prevalence ; Sexual Behavior ; Substance Abuse, Intravenous ; virology
7.Study on inhibitory effect of different extract fractions from longdan xiegan decoction on biofilms of Candida albicans.
Li-Hua YU ; Ke-Qiao LU ; Gao-Xiang SHI ; Yuan-Yuan YAN ; Liang HE ; Jing SHAO ; Tian-Ming WANG ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2014;39(7):1280-1284
OBJECTIVETo observe the inhibitory effect of different extract fractions from Longdan Xiegan decoction on biofilms of Candida albicans, and discuss its possible mechanism.
METHODThe micro-dilution method and the XTT reduction assay were adopted to explore the antifungal activity of different extract fractions from Longdan Xiegan decoction and detect the inhibitory effect of different extracts on biofilms of C. albicans. The expression quantity of the adhesion related gene ALS1 and hypha formation SUN41 were detected by qRT-PCR.
RESULTThe MICs of extracts from Longdan Xiegan decoction, including petroleum ether, water, butanol, methanol and ethyl acetate, against C. albicans were > 1 000, > 1 000, > 1 000, 125, 125 mg x L(-1). The SMIC50 against biofilms of C. albicanswere > 1 000, > 1000, > 1 000, 500, 500 mg x L(-1). The SMIC50 were > 1 000, > 1 000, > 1 000, > 1 000 and 1 000 mg x L(-1). 1 000 mg x L(-1) ethyl acetate extracts could considerably inhibit the expression of the adhesion related gene ALS1 and hypha formation SUN41.
CONCLUSIONThe ethyl acetate extract showed the greatest activity against Candida albicans biofilms.
Antifungal Agents ; pharmacology ; Biofilms ; drug effects ; growth & development ; Candida albicans ; drug effects ; growth & development ; Candidiasis ; drug therapy ; microbiology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Hyphae ; drug effects ; growth & development ; Microbial Sensitivity Tests
8.Study on norcantharidin-induced apoptosis in SMMC-7721 cells through mitochondrial pathways.
Xian-qian LI ; Shi-he SHAO ; Gui-lian FU ; Xiao-hong HAN ; Hong GAO
Chinese journal of integrative medicine 2010;16(5):448-452
OBJECTIVETo investigate the mechanism of norcantharidin (NCTD)-induced SMMC-7721 hepatoma cell apoptosis.
METHODSSMMC-7721 cell growth inhibition was measured by the MTT method. Apoptosis was detected by Annexin V/propidium iodide staining. The mitochondrial membrane potential was measured by flow cytometry. Western blot analysis was used to evaluate the level of cytochrome c, caspase-3, AIF, Bcl-2 and Bax expression.
RESULTSNCTD inhibited SMMC-7721 cell growth in a time- and dose-dependent manner. The cells treated with NCTD showed the loss of mitochondrial membrane potential. The activities of caspase-3, cytochrome c, AIF, and Bax were up-regulated after NCTD treatment at different doses. The expression of Bcl-2 was decreased after treatment with NCTD.
CONCLUSIONSNCTD could induce SMMC-7721 cell apoptosis. The activation of the mitochondrial pathway was involved in the process of NCTD-induced SMMC-7721 cell apoptosis.
Apoptosis ; drug effects ; Apoptosis Inducing Factor ; metabolism ; Blotting, Western ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cytochromes c ; metabolism ; Flow Cytometry ; Humans ; Liver Neoplasms ; enzymology ; metabolism ; pathology ; Membrane Potentials ; drug effects ; Mitochondria ; drug effects ; metabolism ; bcl-2-Associated X Protein ; metabolism
9.Changes of NF-kappaB/I kappa B alpha in N-methyl-N-nitrosourea-induced retinal damage in rats.
Jin-nan YANG ; He-qin ZHAN ; Jin-mao CHEN ; Shao-chun LIN ; Dai LI ; Shi-xing HU
Journal of Southern Medical University 2006;26(5):603-605
OBJECTIVETo observe the changes of nuclear factor-kappa B (NF-kappaB) in the course of N-methyl-N-nitrosourea (MNU)-induced apoptosis of rat retinal photoreceptor cells and investigate the mechanism of MNU-induced retinal damage.
METHODSA single intraperitoneal injection of 60 mg/kg MNU was given to 50-day-old female rats, which were sacrificed at different intervals after MNU treatment. The retinal damage was examined with optical microscopy and photoreceptor cell apoptosis detected by TUNEL assay. Western blotting was performed to analyze the changes in NF-kappaB.
RESULTSPyknosis of the photoreceptor cell nuclei and disorientation of the outer segment of the photoreceptor layer was observed 24 h after MNU treatment, and the outer nuclear layer and photoreceptor layer were almost completely lost on day 7. Photoreceptor cell apoptosis peaked at 24 h, and in the apoptotic cascade, NF-kappaB p65 protein was only detected 12 and 24 h after MNU treatment, whereas the amount of I kappa B alpha, in contrast, markedly increased in the cytoplasm as well as in the nuclei.
CONCLUSIONMNU-induced retinal damage might be mediated through the signaling pathway of NF-kappaB/I kappa B alpha.
Animals ; Apoptosis ; drug effects ; Blotting, Western ; Female ; I-kappa B Proteins ; metabolism ; In Situ Nick-End Labeling ; Methylnitrosourea ; toxicity ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Retinal Diseases ; chemically induced ; metabolism ; pathology
10.The detection and significance of high mobility group box chromosomal protein 1, RORγt and inter leukin-17 in peripheral blood of rheumatoid arthritis
Yan SHI ; Shengjun WANG ; Jianguo CHEN ; Yuan XUE ; Zhiqiang HE ; Chenglin ZHOU ; Dong ZHENG ; Heng YANG ; Yazhen LI ; Jia TONG ; Zhaoliang SU ; Qixiang SHAO ; Huaxi XU
Chinese Journal of Rheumatology 2010;14(3):147-150
Objective To detect the expression levels of high mobility group box chromosomal protein 1 (HMGB1) and Th17 cells transcription factors, related cytokines in peripheral blood of rheumatoid arthritis (RA) patients and analyze the relations between HMGB1 and CRP, ESR, RF in RA patients. The other aim of this study is to identify the expression level of HMGBI and the relationship between HMGB1 and Th17 in RA patients. Methods The mRNA levels of HMGB1, RORyt, interleukin (IL)-17 in the peripheral blood mononuclear cells (PBMC) were determined by quantitative real-time PCR (QRT-PCR) from 80 patients with rheumatoid arthritis,including 32 RA patients in stable phase and 48 patients in active phase, and 50 healthy volunteers. The concentration of HMGB1, IL-23, IL-17 in plasma were detected by enzyme linked immunosorbent assay (ELISA), one-way ANOVA and Spearman's correleation were adopted for statistical analysis.Results The mRNAs of HMGBI, RORyt and IL-17 in RA patients were higher than that in healthy control group (P<0.05), especially in active RA patients [ HMGB 1 (0.424±0.262) pg/ml, RORγt (0.34±0.25) pg/ml,IL-17 (1.42±0.38) pg/ml,P<0.01 ] when compared with patients with stable disease. The concentration of HMGB1, IL-23 and IL-17 in the plasma of RA patients was higher than that of the healthy control group (P< 0.05), and was positively correlated with the expression levels of HMGB1, Th 17-associated factors and the level of CRP, ESR, RF in RA patients' plasma(P<0.05). Conclusion The HMGB1 and Thl7 cells levels are higher in active RA patients than those in patients with stable disease, arid there is significant positive correlation between them. Detection of peripheral HMGB1 and Thl7 cell-specific transcription factors or related cytokines can help to understand the development and progress of rheumatoid arthritis and provide clues for new treatment targets for RA.