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Production of Tween 80-inducing Esterase by Acinetobacter sp. B1 Using Response Surface Methodology

  • Ma, Peiyu;Li, Yuqi;Miao, Chensi;Sun, Yunpeng;Liu, Chunhui;Li, Huijuan
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.87-95
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    • 2019
  • Esterase produced by Acinetobacter sp. B1 (strain B1) was optimized by means of one-variable-at-a-time and response surface methodologies. Results of the one-variable-at-a-time experiment showed that Tween 80 significantly increased esterase production of strain B1. The addition of Tween 80 to the culture medium increased the biomass and esterase activity of strain B1, stimulated content of total extracellular protein, and enhanced the oleic acid (C18:1) composition in the cell membrane of strain B1. The influence of eight culture variables on esterase production was evaluated by Plackett-Burman design. Results showed that Tween 80, pH, and $K_2HPO_4$ significantly affected the esterase production of strain B1. Tween 80, pH, and $K_2HPO_4$ were further optimized by central composite design. Under the optimized conditions (w/v, soluble starch 2.5%, tryptone 1.5%, Tween 80 0.8%, $K_2HPO_4$ 0.5%, NaCl 0.5%, pH 8.0, inoculum size 1%, and inoculum age 24 h), the maximum esterase activity of strain B1 was 152.13 U/ml, which was 10-fold higher than that of non-optimization after 36 h cultivation.

Complete Genome Sequencing of Bacillus velezensis WRN014, and Comparison with Genome Sequences of other Bacillus velezensis Strains

  • Wang, Junru;Xing, Juyuan;Lu, Jiangkun;Sun, Yingjiao;Zhao, Juanjuan;Miao, Shaohua;Xiong, Qin;Zhang, Yonggang;Zhang, Guishan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.794-808
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    • 2019
  • Bacillus velezensis strain WRN014 was isolated from banana fields in Hainan, China. Bacillus velezensis is an important member of the plant growth-promoting rhizobacteria (PGPR) which can enhance plant growth and control soil-borne disease. The complete genome of Bacillus velezensis WRN014 was sequenced by combining Illumina Hiseq 2500 system and Pacific Biosciences SMRT high-throughput sequencing technologies. Then, the genome of Bacillus velezensis WRN014, together with 45 other completed genome sequences of the Bacillus velezensis strains, were comparatively studied. The genome of Bacillus velezensis WRN014 was 4,063,541bp in length and contained 4,062 coding sequences, 9 genomic islands and 13 gene clusters. The results of comparative genomic analysis provide evidence that (i) The 46 Bacillus velezensis strains formed 2 obviously closely related clades in phylogenetic trees. (ii) The pangenome in this study is open and is increasing with the addition of new sequenced genomes. (iii) Analysis of single nucleotide polymorphisms (SNPs) revealed local diversification of the 46 Bacillus velezensis genomes. Surprisingly, SNPs were not evenly distributed throughout the whole genome. (iv) Analysis of gene clusters revealed that rich gene clusters spread over Bacillus velezensis strains and some gene clusters are conserved in different strains. This study reveals that the strain WRN014 and other Bacillus velezensis strains have potential to be used as PGPR and biopesticide.

Characterization and Antioxidant Activity of Released Exopolysaccharide from Potential Probiotic Leuconostoc mesenteroides LM187

  • Zhang, Qing;Wang, Jie;Sun, Qing;Zhang, Shu-Ming;Sun, Xiang-Yang;Li, Chan-Yuan;Zheng, Miao-Xin;Xiang, Wen-Liang;Tang, Jie
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1144-1153
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    • 2021
  • A released exopolysaccharide (rEPS)-producing strain (LM187) with good acid resistance, bile salt resistance, and cholesterol-lowering properties was isolated from Sichuan paocai and identified as Leuconostoc mesenteroides subsp. mesenteroides. The purified rEPS, designated as rEPS414, had a uniform molecular weight of 7.757 × 105 Da. Analysis of the monosaccharide composition revealed that the molecule was mainly composed of glucose. The Fourier transform-infrared spectrum showed that rEPS414 contained both α-type and β-type glycosidic bonds. 1H and 13C nuclear magnetic resonance spectra analysis showed that the purified rEPS contained arabinose, galactose, and rhamnose, but less uronic acid. Scanning electron microscopy demonstrated that the exopolysaccharide displayed a large number of scattered, fluffy, porous cellular network flake structures. In addition, rEPS414 exhibited strong in vitro antioxidant activity. These results showed that strain LM187 and its rEPS are promising probiotics with broad prospects in industry.

Sitagliptin attenuates endothelial dysfunction independent of its blood glucose controlling effect

  • Chang, Xin-Miao;Xiao, Fei;Pan, Qi;Wang, Xiao-Xia;Guo, Li-Xin
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.5
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    • pp.425-437
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    • 2021
  • Although the contributions of sitagliptin to endothelial dysfunction in diabetes mellitus were previously reported, the mechanisms still undefined. Autophagy plays an important role in the development of diabetes mellitus, but its role in diabetic macrovascular complications is unclear. This study aims to observe the effect of sitagliptin on macrovascular endothelium in diabetes and explore the role of autophagy in this process. Diabetic rats were induced through administration of high-fat diet and intraperitoneal injection of streptozotocin. Then diabetic rats were treated with or without sitagliptin for 12 weeks. Endothelial damage and autophagy were measured. Human umbilical vein endothelial cells were cultured either in normal glucose or in high glucose medium and intervened with different concentrations of sitagliptin. Rapamycin was used to induce autophagy. Cell viability, apoptosis and autophagy were detected. The expressions of proteins in c-Jun N-terminal kinase (JNK)-Bcl-2-Beclin-1 pathway were measured. Sitagliptin attenuated injuries of endothelium in vivo and in vitro. The expression of microtubuleassociated protein 1 light chain 3 II (LC3II) and beclin-1 were increased in aortas of diabetic rats and cells cultured with high-glucose, while sitagliptin inhibited the over-expression of LC3II and beclin-1. In vitro pre-treatment with sitagliptin decreased rapamycin-induced autophagy. However, after pretreatment with rapamycin, the protective effect of sitagliptin on endothelial cells was abolished. Further studies revealed sitagliptin increased the expression of Bcl-2, while inhibited the expression of JNK in vivo. Sitagliptin attenuates injuries of vascular endothelial cells caused by high glucose through inhibiting over-activated autophagy. JNK-Bcl-2-Beclin-1 pathway may be involved in this process.

A comprehensively overall track-bridge interaction study on multi-span simply supported beam bridges with longitudinal continuous ballastless slab track

  • Su, Miao;Yang, Yiyun;Pan, Rensheng
    • Structural Engineering and Mechanics
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    • v.78 no.2
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    • pp.163-174
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    • 2021
  • Track-bridge interaction has become an essential part in the design of bridges and rails in terms of modern railways. As a unique ballastless slab track, the longitudinal continuous slab track (LCST) or referred to as the China railway track system Type-II (CRTS II) slab track, demonstrates a complex force mechanism. Therefore, a comprehensive track-bridge interaction study between multi-span simply supported beam bridges and the LCST is presented in this work. In specific, we have developed an integrated finite element model to investigate the overall interaction effects of the LCST-bridge system subjected to the actions of temperature changes, traffic loads, and braking forces. In that place, the deformation patterns of the track and bridge, and the distributions of longitudinal forces and the interfacial shear stress are studied. Our results show that the additional rail stress has been reduced under various loads and the rail's deformation has become much smoother after the transition of the two continuous structural layers of the LCST. However, the influence of the temperature difference of bridges is significant and cannot be ignored as this action can bend the bridge like the traffic load. The uniform temperature change causes the tensile stress of the concrete track structure and further induce cracks in them. Additionally, the influences of the friction coefficient of the sliding layer and the interfacial bond characteristics on the LCST's performance are discussed. The systematic study presented in this work may have some potential impacts on the understanding of the overall mechanical behavior of the LCST-bridge system.

Inclusion and mechanical properties of ODS-RAFM steels with Y, Ti, and Zr fabricated by melting

  • Qiu, Guo-xing;Wei, Xu-li;Bai, Chong;Miao, De-jun;Cao, Lei;Li, Xiao-ming
    • Nuclear Engineering and Technology
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    • v.54 no.7
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    • pp.2376-2385
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    • 2022
  • Two groups of oxide dispersion-strengthened reduced-activation ferritic/martensitic steels (A and B) were prepared by adding Y, Ti, and Zr into steels through vacuum induction melting to investigate the inclusions, microstructures, mechanical properties of the alloys. Results showed that particles with Y, Ti, and Zr easily formed. Massive, Zr-rich inclusions were found in B steel. Density of micron inclusions in A steel was 1.42 × 1014 m-3, and density of nanoparticles was 3.61 × 1016 m-3. More and finer MX carbides were found in steel tempered at 650 ℃, and yield strengths (YS) of A and B steel were 714±2 and 664±3.5 MPa. Thermomechanical processing (TMP) retained many dislocations, which improved the mechanical properties. YSs of A and B treated by TMP were 725±3 and 683±4 MPa. The existence of massive Zr-rich inclusions in B steels interrupted the continuity of the matrix and produced microcracks (fracture), which caused a reduction in mechanical properties. The presence of fine prior austenite grain size and inclusions was attributed to the low DBTTs of the A steels; DBTTs of A650 and A700 alloy were -79 and -65 ℃. Tempering temperature reduction and TMP are simple, readily useable methods that can lead to a superior balance of strength and impact toughness in industry applications.

Influencing Factors of High PTSD Among Medical Staff During COVID-19: Evidences From Both Meta-analysis and Subgroup Analysis

  • Qi, Guojia;Yuan, Ping;Qi, Miao;Hu, Xiuli;Shi, Shangpeng;Shi, Xiuquan
    • Safety and Health at Work
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    • v.13 no.3
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    • pp.269-278
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    • 2022
  • Background: PTSD (Post-traumatic stress disorder, PTSD) had a great impact on health care workers during the COVID-19 (Corona Virus Disease 2019, COVID-19). Better knowledge of the prevalence of PTSD and its risk factors is a major public health problem. This study was conducted to assess the prevalence and important risk factors of PTSD among medical staff during the COVID-19. Methods: The databases were searched for studies published during the COVID-19, and a PRISMA (preferred reporting items for systematic review and meta-analysis) compliant systematic review (PROSPERO-CRD 42021278970) was carried out to identify articles from multiple databases reporting the prevalence of PTSD outcomes among medical staff. Proportion random effect analysis, I2 statistic, quality assessment, subgroup analysis, and sensitivity analysis were carried out. Results: A total of 28 cross-sectional studies and the PTSD results of doctors and nurses were summarized from 14 and 27 studies: the prevalences were 31% (95% CI [confidence interval, CI]: 21%-40%) and 38% (95% CI: 30%-45%) in doctors and nurses, respectively. The results also showed seven risks (p < 0.05): long working hours, isolation wards, COVID-19 symptoms, nurses, women, fear of infection, and pre-existing mental illness. Two factors were of borderline significance: higher professional titles and married. Conclusion: Health care workers have a higher prevalence of PTSD during COVID-19. Health departments should provide targeted preventive measures for medical staff away from PTSD.

Genome-wide identification of long noncoding RNA genes and their potential association with mammary gland development in water buffalo

  • Jin, Yuhan;Ouyang, Yina;Fan, Xinyang;Huang, Jing;Guo, Wenbo;Miao, Yongwang
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1656-1665
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    • 2022
  • Objective: Water buffalo, an important domestic animal in tropical and subtropical regions, play an important role in agricultural economy. It is an important source for milk, meat, horns, skin, and draft power, especially its rich milk that is the great source of cream, butter, yogurt, and many cheeses. In recent years, long noncoding RNAs (lncRNAs) have been reported to play pivotal roles in many biological processes. Previous studies for the mammary gland development of water buffalo mainly focus on protein coding genes. However, lncRNAs of water buffalo remain poorly understood, and the regulation relationship between mammary gland development/milk production traits and lncRNA expression is also unclear. Methods: Here, we sequenced 22 samples of the milk somatic cells from three lactation stages and integrated the current annotation and identified 7,962 lncRNA genes. Results: By comparing the lncRNA genes of the water buffalo in the early, peak, and late different lactation stages, we found that lncRNA gene lnc-bbug14207 displayed significantly different expression between early and late lactation stages. And lnc-bbug14207 may regulate neighboring milk fat globule-EGF factor 8 (MFG-E8) and hyaluronan and proteoglycan link protein 3 (HAPLN3) protein coding genes, which are vital for mammary gland development. Conclusion: This study provides the first genome-wide identification of water buffalo lncRNAs and unveils the potential lncRNAs that impact mammary gland development.

Nitrogen metabolism and mammary gland amino acid utilization in lactating dairy cows with different residual feed intake

  • Xie, Yunyi;Miao, Chao;Lu, Yi;Sun, Huizeng;Liu, Jianxin
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1600-1606
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    • 2021
  • Objective: This study was conducted to enhance our understanding of nitrogen (N) metabolism and mammary amino acid (AA) utilization in lactating cows with divergent phenotypes of residual feed intake (RFI). Methods: Fifty-three multiparous mid-lactation Holstein dairy cows were selected for RFI measurements over a 50-d experimental period. The 26 cows with the most extreme RFI values were classified into the high RFI (n = 13) and low RFI (n = 13) groups, respectively, for analysis of N metabolism and AA utilization. Results: Compared with the high RFI cows, the low RFI animals had lower dry matter intake (p<0.01) with no difference observed in milk yield between the two groups (p>0.10). However, higher ratios of milk yield to dry matter intake (p<0.01) were found in the low RFI cows than in the high RFI cows. The low RFI cows had significant greater ratios of milk protein to metabolizable protein (p = 0.02) and milk protein to crude protein intake than the high RFI cows (p = 0.01). The arterial concentration and mammary uptake of essential AA (p<0.10), branched-chain AA (p<0.10), and total AA (p<0.10) tended to be lower in the low RFI cows. Additionally, the low RFI cows tended to have a lower ratio of AA uptake to milk output for essential AA (p = 0.08), branched-chain AA (p = 0.07) and total AA (p = 0.09) than the high RFI cows. Conclusion: In summary, both utilization of metabolizable protein for milk protein and mammary AA utilization are more efficient in cows with lower RFI than in the high RFI cows. Our results provide new insight into the protein metabolic processes (related to N and AA) involved in feed efficiency.

Computerized bone age estimation system based on China-05 standard

  • Yin, Chuangao;Zhang, Miao;Wang, Chang;Lin, Huihui;Li, Gengwu;Zhu, Lichun;Fei, Weimin;Wang, Xiaoyu
    • Advances in nano research
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    • v.12 no.2
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    • pp.197-212
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    • 2022
  • The purpose of this study is to develop an automatic software system for bone age evaluation and to evaluate its accuracy in testing and feasibility in clinical practice. 20394 left-hand radiographs of healthy children (2-18 years old) were collected from China Skeletal Development Survey data of 1998 and China Skeletal Development Survey data of 2005. Three experienced radiologists and China-05 standard maker jointly evaluate the stages of bone development and the reference bone age was determined by consensus. 1020 from 20394 radiographs were picked randomly as test set and the remaining 19374 radiographs as training set and validation set. Accuracy of the automatic software system for bone age assessment is evaluated in test set and two clinical test sets. Compared with the reference standard, the automatic software system based on RUS-CHN for bone age assessment has a 0.04 years old mean difference, ±0.40 years old in 95% confidence interval by single reading, a 85.6% percentage agreement of ratings, a 93.7% bone age accuracy rate, 0.17 years old of MAD, 0.29 years old of RMS; Compared with the reference standard, the automatic software system based on TW3-C RUS has a 0.04 years old mean difference, a ±0.38 years old in 95% confidence interval by single reading, a 90.9% percentage agreement of ratings, a 93.2% bone age accuracy rate, a 0.16 years of MAD, and a 0.28 years of RMS. Automatic software system, AI-China-05 showed reliably accuracy in bone age estimation and steady determination in different clinical test sets.