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        Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

        ( Lintao Hu ),( Jun Huang ),( Hui Li ),( Yao Jin ),( Chongde Wu ),( Rongqing Zhou ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.11

        Fluorescence in-situ hybridization (FISH) is a common and popular method used to investigate microbial communities in natural and engineered environments. In this study, two specific 16S rRNA-targeted oligonucleotide probes, CLZ and KCLZ, were designed and verified to quantify the genus Clostridium and the species Clostridium kluyveri. The optimal concentration of hybridization buffer solution for both probes was 30% (w/v). The specificity of the designed probes was high due to the use of pellets from pure reference strains. Feasibility was tested using samples of Chinese liquor from the famed Luzhou manufacturing cellar. The effectiveness of detecting target cells appears to vary widely in different environments. In pit mud, the detection effectiveness of the target cell by probes CLZ and KCLZ was 49.11% and 32.14%, respectively. Quantitative analysis by FISH technique of microbes in pit mud and fermented grains showed consistency with the results detected by qPCR and PCR-DGGE techniques, which showed that the probes CLZ and KCLZ were suitable to analyze the biomass of Clostridium spp. and C. kluyveri during liquor fermentation. Therefore, this study provides a method for quantitative analysis of Clostridium spp. and C. kluyveri and monitoring their community dynamics in microecosystems.

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        Systemic Reactions to Dust Mite Subcutaneous Immunotherapy: A 3-Year Follow-up Study

        Xiang Dong,Nan Huang,Wenjing Li,Lintao Hu,Xiaolong Wang,Yin Wang,Ning Xiang,Guanghui Liu,Rongfei Zhu 대한천식알레르기학회 2016 Allergy, Asthma & Immunology Research Vol.8 No.5

        Purpose: The incidence of allergen specific immunotherapy-related systemic reactions (SRs) varies among different studies, and many factors are likely to contribute to SRs. This study aims to investigate the incidence, characteristics, and risk factors of SRs to standardize dust mite-specific subcutaneous immunotherapy (SCIT) in Central China. Methods: All patients receiving standardized dust mites (100-100,000 SQ-U/mL; Alutard SQ, Hørsholn, Denmark) immunotherapy were followed up. Recorded data included demographics, diagnosis, patient status, pulmonary function testing results before and after each injection, allergen dosage, and details of SRs. Results: From June 2011 to August 2014, a total of 208 patients received 4,369 injections; 27 (13.0%) patients experienced 48 (1.1%) systemic reactions. Most of the SRs were grade 2 reactions (n=30, 62.5%), followed by grade 1 (n=11, 22.9%), grade 3 (n=7, 14.6%), and no fatal reactions occurred. Forty-six SRs (95.8%) occurred within 30 minutes. Higher SR rates were associated with high concentration extracts (100,000 SQ-U/mL), injections with concomitant local reactions (LRs), children, asthma and high sensitivity (skin prick test 3+/4+ and/or sIgE≥17.5 kUA/L) (P<0.05). The estimated odds of SRs increased in children (OR=6.57; 95% CI: 1.88-22.97, P=0.003), asthmatic patients (OR=4.10; 95% CI: 1.72-9.80, P=0.002), and injections with LRs (OR=2.41; 95% CI: 1.33-4.36, P=0.004).Conclusions: The incidence of SRs to dust mite SCIT was low, and multiple factors were associated with the increased incidence of SRs. Children, asthmatics and patients with concomitant LR may be prone to develop SRs.

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        Bias load analysis and asymmetric modification of rollers in the main drive bearing of large roadheader

        Xin-Qi Wang,Wei Sun,Lintao Wang,Shi-Hu Liang,Xiao-Kai Mu 대한기계학회 2023 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.37 No.6

        An asymmetric modification method for the roller profile was proposed to address the bias load effect of rollers in the main drive bearing of a large roadheader. A contact mechanics model of the bearing was solved, and line contact theory was used to explore the effect of combined load and convexity on the bias load effect. The contact stress was found to be asymmetrically distributed along the contact line. This trend tended to be remarkable with the decrease in axial load or the increase in overturning moment. Symmetrically modified rollers can be used to improve the bias load effect by increasing convexity. However, this process led to the contact stress concentrating in the roller center. The influence of the design parameters for asymmetric modification on the roller profile and the contact stress was considered. The ideal design parameters were determined by using a genetic algorithm. The contact condition and fatigue life were remarkably increased.

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