Characterization of Enterobacter cloacae under Phoxim Stress by Two-dimensional Gel Electrophoresis

Dijun Zhang,Jiajie Xu,Weina He,Qianqian Tong,Liping Chen,Jun Zhou,Xiurong Su 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.3

Phoxim, an organophosphorus pesticide, was widely used in agriculture but was banned because of its associated toxic runoff, which accumulated in surrounding water outlets. Here, Enterobacter cloacae was isolated from water outlets adjacent to industrial, agricultural and residential sewage having high organic phosphorus contents. To investigate the ability of E. cloacae to respond and survive under phoxim stress, both at the proteomic and transcriptional level, two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS) and quantitative real-time PCR (qPCR) were used. A 2-DE analysis showed that 12 h post phoxim exposure, 46 proteins were differentially expressed and 21 protein spots were selected for MALDI-TOF-MS/MS identification. This resulted in the successful identification of 16 spots, nine up-regulated and seven down-regulated. Of these, stress response proteins, including the chaperone protein DnaK, GroEL, F0F1 ATP synthase subunit beta, phosphoenolpyruvate carboxykinase and outer membrane channel protein, were only expressed in the phoxim-treated experimental group. To validate these findings, seven genes were selected and positively confirmed using qPCR. The results suggest that the identified expression patterns relate to E. cloacae resistance and tolerance to high phoxim concentrations.

Transverse Flexural Behaviour of Steel-Engineering Cementitious Composites (ECC) Composite Deck under Negative and Positive Bending Forces

Yanhua Guan,Jiajie Wu,Renjuan Sun,Hongzhi Zhang,Yanqiu Hu,Fei Wang 대한토목학회 2021 KSCE JOURNAL OF CIVIL ENGINEERING Vol.25 No.8

Orthotropic steel bridge deck system usually consists of an orthotropic steel deck and an asphalt overlay. Fatigue cracks of the orthotropic steel deck and premature damage of the asphalt overlay are frequently reported for such system. Engineering cementitious composites (ECC) was therefore proposed to replace the asphalt overlay to address the aforementioned issues. The current study presents an investigation on the transversal flexural behaviour of the ECC under bending forces. Influence of the bending force direction, cover thickness, the number of longitudinal reinforcement steel bars on the flexural performance was revealed. Responses with regard to the load-deflection curve, failure mode, the ultimate capacity, the cracking behaviour, the interfacial slip between ECC and steel deck and the nominal cracking stresses were analysed. The results showed that the load-deflection response under both negative and positive bending forces exhibits elastic stage, crack-developing stage and yield stage. Compared with the reinforcement ratio, cover thickness plays a more significant role on the flexural performances. When the cover thickness decreases from 35 mm to 25 mm, the peak load increases by 21% − 25%. ECC maintains its feature of being ductile, with high tensile and compressive strain capacity in the composite slab. From the design point of view, reducing the cover thickness and increasing the reinforcement ratio can improve the ultimate load and cracking stress, and reduce the internal slip, strain and crack width of the composite slab. It is expected that the current study can provide basic knowledge to the design and application of the steel-ECC composite deck system.

The polymorphisms of bovine PCSK1 gene and their associations with growth traits

Limin Shan,Jiajie Sun,Chunlei Zhang,Xingtang Fang,Chuzhao Lei,Xianyong Lan,Hong Chen 한국유전학회 2011 Genes & Genomics Vol.33 No.1

Proprotein convertase 1 (PCSK1) is an endopeptidase involved in proteolytic processing of peptide hormone precursors in granules of the regulated secretory pathway of endocrine cells and mutations in PCSK1 gene are thought to cause obesity. In the present study, based on PCR-SSCP and DNA sequencing methods, polymorphisms of the PCSK1 gene were detected in 858 individuals from five breeds (Nanyang cattle, Qinchuan cattle, Jiaxian cattle, Luxi cattle and Chinese Holstein). The results showed that only P_8 locus showed polymorphisms and 3 synonymous SNPs of PCSK1 gene were identified. Additionally, significant statistical difference was found in bovine birth weight and diplotype MM were 7.35% higher than diplotype XY.

High-Pitch Coronary CT Angiography at 70 kVp Adopting a Protocol of Low Injection Speed and Low Volume of Contrast Medium

Ruiqi Feng,Jiajie Tong,Xiaofei Liu,Yu Zhao,Liang Zhang 대한영상의학회 2017 Korean Journal of Radiology Vol.18 No.5

Objective: To evaluate the feasibility and image quality (IQ) of prospectively high-pitch coronary CT angiography (CCTA) with low contrast medium injection rate at 70 kVp. Materials and Methods: One hundred and four patients with suspected coronary artery disease (body mass index < 26 kg/m2, sinus rhythm and heart rate < 70 beats/min) were prospectively enrolled and randomly divided into two groups. In group A and group B, 28 mL and 40 mL of 370 mgI/mL iodinated contrast media was administrated at a flow rate of 3.5 and 5 mL/s, respectively. CT values, noise, signal-to-noise ratio, contrast-to-noise ratio (CNR) of the proximal segments of coronary arteries and subjective IQ were evaluated. Results: The CT values and noise in group A were significantly lower than those in group B (434–485 Hounsfield units [HU] vs. 772–851 HU, all p < 0.001; 17.8–22.3 vs. 23.3–26.4, all p < 0.005). The CNRs of the right coronary artery and left main artery showed no statistical difference between the two groups (42.1 ± 13.8 vs. 36.8 ± 16.0, p = 0.074; 38.7 ± 10.6 vs. 38.1 ± 17.0, p = 0.819). No statistical difference was observed between the two groups in IQ scores (3.04 ± 0.75 vs. 3.0 ± 0.79, p = 0.526) and diagnostic ratio (96.1% [50/52] vs. 94.2% [49/52], p = 0.647). Conclusion: Prospective high-pitch CCTA at 70 kVp with 28 mL of contrast media and injection rate of 3.5 mL/s could provide diagnostic IQ for normal-weight patients with heart rate of < 70 beats/min.

Fluctuation Power Control Strategy for MMC-based SST to Reduce the Submodule Capacitor Voltage Oscillation

Xiaolong Cai,Jianwen Zhang,Gang Shi,Xu Cai,Jianqiao Zhou,Zhuyong Li,Jiacheng Wang,Jiajie Zang 전력전자학회 2019 ICPE(ISPE)논문집 Vol.2019 No.5

In the future AC-DC hybrid distribution system, solid state transformer (SST) will take place of conventional transformer, as it could reduce the weight and volume of transformer, and achieve voltage and current regulation, power flow control, power quality enhancement. MMC-based SST is a novel SST topology, which could provide the MVDC port. However, the excessive bulky submodule capacitors limits this topology into practical application. This paper proposes a fluctuation power control strategy for MMC-based SST to reduce the submodule capacitor voltage oscillation, which could optimize the capacitance and reduce the volume and cost of capacitor. The feasibility of this proposed method is verified by MATLAB simulation.

Effects of a novel marine natural product: pyrano indolone alkaloid fibrinolytic compound on thrombolysis and hemorrhagic activities in vitro and in vivo

Ting Yan,Wen-Hui Wu,Tongwei Su,Jiajie Chen,Quangang Zhu,Chaoyan Zhang,Xiaoyu Wang,Bin Bao 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.8

Fungi fibrinolytic compound 1 (FGFC1) is a novel marine natural product as a low-weight fibrinolytic pyranoindole molecule, whose thrombolytic effects were evaluated on FITC-fibrin (Fluorescein isothiocyanate, FITC) degradation methods in vitro and on acute pulmonary thromboembolism animal model in vivo. We determined the FGFC1 induced thrombolysis that stems from its fibrin(ogen)olytic activities as measured by fibrin(ogen) degradation products (FDPs) experiment, acute pulmonary thromboembolism animal model experiment, and euglobulin lysis assay. In vitro, measurement of FITCfibrin degradation revealed that fibrin hydrolysis occurred in a concentration-dependent manner of FGFC1 from 5 to 25 l mol/L. In vivo test of a classical acute pulmonary thromboembolism model in rat showed that when the injected dose was 5 mg/kg or above, FGFC1 was effectivein dissolution of extrinsic FITC-fibrin induced blood clots. Euglobulin lysis time (ELT) in FGFC1-treated rats was shortened 30 s compared with rats in the positive control group, which were injected with clopidogrel sulfate and single-chain urokinase-type plasminogen activator. As compared to the control, FGFC1 (5–25 mg/kg) did not significantly alter the formation of fibrinogen and FDPs in vivo. Our research indicates that FGFC1 presents pharmacodynamic action in both the thrombolysis and the hemolytic procedure, which can be characterized by fibrinogenolysis in blood and FDPs in plasma. In vivo, increasing fibrinolytic doses of FGFC1 from 5 to 25 mg/kg did not induce fibrinogenolysis when compared with control group, this result corresponds to that FGFC1 did not induce the increasing of FDPs (compared with the salinetreated control). It indicates that the FGFC1 may act as a novel thrombolytic agent and represent an effective approach to the treatment of thrombus without significant risk of hemorrhagic activity.

Phosphatidylcholine regulates NF-κB activation in attenuation of LPS-induced inflammation: evidence from in vitro study

Meijuan Cheng,Hongying Pan,Yining Dai,Jiajie Zhang,Yongxi Tong,Yicheng Huang,Mingshan Wang,Haijun Huang 한국통합생물학회 2018 Animal cells and systems Vol.22 No.1

Phosphatidylcholine (PC) has been demonstrated as anti-inflammatory and antioxidant/pro-oxidant molecules. In this study, we investigated the protective effects of PC on inflammatory bowel disease (IBD) caused by lipopolysaccharide (LPS)-induced injury in intestinal epithelia cells. The IEC-6 cells (intestinal epithelia cells) were stimulated with LPS (1 μg/mL) for 24 h with or without PC pretreatment, in the next steps: (1) the level of the inflammatory cytokine tumor necrosis factor (TNF)-α was measured with ELISA; (2) the nuclear translocation and phosphorylation of NF-κB was investigated with Western blot, EMSA, immunofluorence assay; (3) the protein phosporylation levels in MAPK signaling pathway were detected with Western blot method. The results showed: (1) compared with the normal group, 10 and 20 μg/mL of PC significantly inhibited the production and activation of TNF-α, (P < 0.01); (2) pretreatment with PC inhibited LPS-induced nuclear translocation and phosphorylation of p65 in IEC-6 cells; (3) pretreatment with PC inhibited the protein phosphorylation levels in MAPK signaling pathway. Our findings indicated that PC had the effect to protect IEC-6 cells from LPS-induced injury and this effect was exerted possibly through inhibiting the TNF-ɑ secretion, down-regulating nuclear translocation and phosphorylation of p65 and inhibiting MAPK signaling pathways.

Phosphatidylcholine regulates NF-κB activation in attenuation of LPS-induced inflammation: evidence from in vitro study

Cheng, Meijuan,Pan, Hongying,Dai, Yining,Zhang, Jiajie,Tong, Yongxi,Huang, Yicheng,Wang, Mingshan,Huang, Haijun 한국통합생물학회 2018 Animal cells and systems Vol.22 No.1

Phosphatidylcholine (PC) has been demonstrated as anti-inflammatory and antioxidant/pro-oxidant molecules. In this study, we investigated the protective effects of PC on inflammatory bowel disease (IBD) caused by lipopolysaccharide (LPS)-induced injury in intestinal epithelia cells. The IEC-6 cells (intestinal epithelia cells) were stimulated with LPS ($1{\mu}g/mL$) for 24 h with or without PC pretreatment, in the next steps: (1) the level of the inflammatory cytokine tumor necrosis factor (TNF)-${\alpha}$ was measured with ELISA; (2) the nuclear translocation and phosphorylation of NF-${\kappa}B$ was investigated with Western blot, EMSA, immunofluorence assay; (3) the protein phosporylation levels in MAPK signaling pathway were detected with Western blot method. The results showed: (1) compared with the normal group, 10 and $20{\mu}g/mL$ of PC significantly inhibited the production and activation of TNF-${\alpha}$, (P < 0.01); (2) pretreatment with PC inhibited LPS-induced nuclear translocation and phosphorylation of p65 in IEC-6 cells; (3) pretreatment with PC inhibited the protein phosphorylation levels in MAPK signaling pathway. Our findings indicated that PC had the effect to protect IEC-6 cells from LPS-induced injury and this effect was exerted possibly through inhibiting the TNF-${\alpha}$ secretion, down-regulating nuclear translocation and phosphorylation of p65 and inhibiting MAPK signaling pathways.

Effects of echinomycin on endothelin-2 expression and ovulation in immature rats primed with gonadotropins

Zhengchao Wang,Yanqing Wu,Liyun Chen,Qianping Luo,Jisen Zhang,Jiajie Chen,Zimiao Luo,Xiaohong Huang,Yong Cheng,Zhenghong Zhang 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.10

Echinomycin is a small-molecule inhibitor of hypoxia- inducible factor-1 DNA-binding activity, which plays a crucial role in ovarian ovulation in mammalians. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1α-mediated endothelin (ET)-2 expressions contributed to ovarian ovulation in response to human chorionic gonadotropin (hCG) during gonadotropin-induced superuvulation. By real-time RT-PCR analysis, ET-2 mRNA level was found to significantly decrease in the ovaries after chinomycin treatment, while HIF-1α mRNA and protein expression was not obviously changed. Further analysis also showed that these changes of ET-2 mRNA were consistent with HIF-1 activity in the ovaires, which is similar with HIF-1α and ET-2 expression in the granulosa cells with gonadotropin and echinomycin treatments. The results of HIF-1α and ET-2 expression in the granulosa cells transfected with cis-element oligodeoxynucleotide (dsODN) under gonadotropin treatment further indicated HIF-1α directly mediated the transcriptional activation of ET-2 during gonadotropin- induced superuvulation. Taken together, these results demonstrated that HIF-1α-mediated ET-2 transcriptional activation is one of the important mechanisms regulating gonadotropin-induced mammalian ovulatory precess in vivo. Echinomycin is a small-molecule inhibitor of hypoxia- inducible factor-1 DNA-binding activity, which plays a crucial role in ovarian ovulation in mammalians. The present study was designed to test the hypothesis that hypoxia-inducible factor (HIF)-1α-mediated endothelin (ET)-2 expressions contributed to ovarian ovulation in response to human chorionic gonadotropin (hCG) during gonadotropin-induced superuvulation. By real-time RT-PCR analysis, ET-2 mRNA level was found to significantly decrease in the ovaries after chinomycin treatment, while HIF-1α mRNA and protein expression was not obviously changed. Further analysis also showed that these changes of ET-2 mRNA were consistent with HIF-1 activity in the ovaires, which is similar with HIF-1α and ET-2 expression in the granulosa cells with gonadotropin and echinomycin treatments. The results of HIF-1α and ET-2 expression in the granulosa cells transfected with cis-element oligodeoxynucleotide (dsODN) under gonadotropin treatment further indicated HIF-1α directly mediated the transcriptional activation of ET-2 during gonadotropin- induced superuvulation. Taken together, these results demonstrated that HIF-1α-mediated ET-2 transcriptional activation is one of the important mechanisms regulating gonadotropin-induced mammalian ovulatory precess in vivo.

De novo genome assembly and annotation of Holothuria scabra (Jaeger, 1833) from nanopore sequencing reads

Luo Honglin,Huang Guanghua,Li Jianbin,Yang Qiong,Zhu Jiajie,Zhang Bin,Feng Pengfei,Zhang Yongde,Yang Xueming 한국유전학회 2022 Genes & Genomics Vol.44 No.12

Background: Holothuria scabra is a costly gourmet and traditional Chinese tonic medicine. However, the lack of high-quality genome information hinders the genetic, phylogenetic, and bioactivator researches. Objective: To construct high-quality genomic data of H. scabra and conduct genome-wide phylogenetic analysis. Methods: The whole genome of a male H. scabra was sequenced based on Nanopore MinION platform, and the sequence was assembled by wtdbg2. Transcriptome sequencing was used to aid the gene annotation. Repeat sequences, non-coding RNA, pseudogene and gene functional annotation were analyzed. 750 single-copy gene families from ten species were applied to construct phylogenetic tree for evolutionary analysis by using the ML method. Results: The H. scabra genome of 1.18 Gb (N50 = 1557,492 bp) with 500.42 Mb of putative repetitive sequences was assembled from a male H. scabra individual, and 16,642 protein-coding genes, 951 pseudogenes, 1791 motifs and 45,400 domains from the generated assembly were identified. The divergence time between H. scabra and its ancestor was estimated approximately 192.6 million years ago. H. scabra and A. japonicas joined together while sea urchin and sea star diverged about 440 Mya ago. Some key genes involved in notochord and gill slits development, skeleton degeneration and nervous system, as well as homeobox genes differ between H. scabra and Apostichopus japonicas. Conclusion: We report the first whole genome of H. scabra with expectation that this will be a valuable resource for genetic, phylogenetic, breading, molecular biology and bioactivator studies of sea cucumbers and other invertebrates.