High-throughput negative detection of SDS-PAGE separated proteins and its application for proteomics

Cong, Wei-Tao,Hwang, Sun-Young,Jin, Li-Tai,He, Hong-Zhang,Choi, Jung-Kap WILEY-VCH Verlag 2010 Electrophoresis Vol.31 No.2

<P>A negative detection method for proteins on SDS-PAGE is described. In this method, Eosin Y (EY) was selectively precipitated in the gel background, which is absent from those zones where proteins are located through the formation of a stable water-soluble protein–dye complex. Negative staining of proteins using EY, allows high-sensitivity, low-cost, and simple protocol. The new described method takes less than an hour to complete all the protocol, with a detection limit of 0.5 ng of single protein band. Comparing with imidazole-zinc negative stain, EY dye provides broader linear dynamic range, higher sensitivity and reproducibility, and better obvious contrast between the protein bands or spots and background. Furthermore, the novel technique developed here presented a real practical method for simultaneous processing of multiple gels, which makes it possible to perform high-throughput staining for proteome research. Additionally, we have also compared the influence of staining method on the quality of mass spectra by PMF.</P>

Numerical study on the cavitating flow through poppet valves concerning the influence of flow instability on cavitation dynamics

Cong Yuan,Lisha Zhu,Shiqi Liu,Du Zunling,He Li 대한기계학회 2022 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.36 No.2

The flow dynamics of the cavitating jet through poppet valves inevitably suffers from the influence of varying openness, but the relevant mechanism remains unclear. Hence, the present study conducts a numerical study on the cavitating flow phenomenon inside poppet valves with two valve seat structures, in order to examine the flow mechanisms underlying varying cavitation phenomenon at different openness. According to the results, the overall cavitation distribution as well as the morphology feature follows a similar pattern due to the same inception mechanism irrelevant of the openness amount. However, examination of the dynamic behaviors confirms discernable variation in 2 coupling effects between flow instability and cavitation. The flow separation at the poppet trailing edge produces non-cavitation flow instability at 0.4 mm openness and attached cavitation at 0.8 mm, which amalgamate with upstream-shed vortex cavitation. The stably attached cavitation within the chamfered groove at 0.4 mm, as the source of non-cavitation flow instability, and the detached cavitation at 0.6 and 0.8 mm openness produce vortex cavitation at the free shearing side. The substantial variation in these two coupling effects contributes sensibly to the increase in the overall cavitation intensity. Meanwhile, the weakened coupling effect between the free shearing vortex and wall shearing vortex, as a consequence of increased potential core thickness at larger openness, leads to the attenuated vortex cavitation at the wall poppet side. Additionally, the vortex cavitation at free shearing side has a larger size in the chamfered valve seat case, due to the coupling of shed flow instability with the Kelvin-Helmholtz instability. As a conclusion, the inception mechanism for the cavitation remains unchanged with openness, while the difference in flow instabilities as well as the associated coupling effects with cavitation contributes to the variation in cavitation intensity.

Schisantherin A Improves Learning and Memory of Mice with D-Galactose-Induced Learning and Memory Impairment Through Its Antioxidation and Regulation of p19/p53/p21/ Cyclin D1/CDK4/RB Gene Expressions

Cong Liu,Weijing Sun,Ning Li,Jiaqi Gao,Chunyan Yu,Chunmei Wang,Jinghui Sun,Shu Jing,JianGuang Chen,He Li 한국식품영양과학회 2018 Journal of medicinal food Vol.21 No.7

Schisantherin A (SCA) was evaluated for possible function in restoring the learning and memory impairment induced by D-galactose in mice. ICR mice were treated with D-galactose subcutaneously (220 mg·kg−1), and followed by SCA in different doses (1.25, 2.50 and 5.00 mg·kg−1, administered orally) for 42 days. Effects of SCA on learning and memory were examined by step-through tests and Morris water maze tests. The activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA) in the peripheral blood and hippocampus of mice were assayed by water-soluble tetrazolium-1 (WST-1) and thiobarbituric acid (TBA) methods. The contents of 8 hydroxy deoxy guanosine (8-OHdG) in the hippocampus of mice were detected by immunosorbent assay methods, respectively. Quantitative real-time PCR and Western Blot were respectively used to detect the expression of p19, p53, p21, cyclin D1, CDK4 and RB genes, and the phosphorylation of RB in the hippocampus of mice. We found that SCA significantly improved the learning and memory impairment induced by D-galactose in mice. After SCA treatment, SOD activity was increased and the content of MDA was decreased in both peripheral blood and hippocampus of mice. 8-OHDG content was also decreased in the hippocampus of mice. Furthermore, the expression of p19, p53 and p21 genes was reduced and the expression of cyclin D1 and CDK4 and the phosphorylation of RB protein were elevated in the hippocampus. SCA may improve the learning and memory impairment induced by D-galactose by enhancing the antioxidant capacity, and regulating the expression of p19/p53/p21/cyclinD1/CDK4 genes, and the phosphorylation of RB protein in the hippocampus of mice.

Disruption of endothelial barrier function is linked with hyposecretion and lymphocytic infiltration in salivary glands of Sjögren's syndrome

Cong, Xin,Zhang, Xue-Ming,Zhang, Yan,Wei, Tai,He, Qi-Hua,Zhang, Li-Wei,Hua, Hong,Lee, Sang-Woo,Park, Kyungpyo,Yu, Guang-Yan,Wu, Li-Ling Elsevier 2018 Biochimica et biophysica acta. Molecular basis of Vol.1864 No.10

<P><B>Abstract</B></P> <P>Sjögren's syndrome (SS) is an inflammatory autoimmune disease that causes hyposecretion in salivary glands. Endothelial tight junctions (TJs) play crucial roles in salivation and barrier function of blood vessels. However, whether the alteration of endothelial TJs were involved in pathogenesis of SS was still unknown. Here, the ultrastructure and function of endothelial TJs in submandibular glands (SMGs) were detected by transmission electron microscopy and in vivo paracellular permeability assay in different aged NOD mouse model for SS. CFSE-labeled lymphocytes were injected into tail vein to trace the infiltration, while claudin-5 expression and distribution were detected by immunofluorescence, qRT-PCR, and western blot. Results showed that the stimulated salivary flow rate was gradually decreased and lymphocytic infiltration was found as age increased in 12- and 21-week-old NOD mice, but not 7-week-old NOD mice. Blood vessels were dilated, while endothelial TJ width and paracellular tracer transport were increased in 12-week-old NOD mice. Moreover, the injected CFSE-labeled lymphocytes were observed in SMGs of 12-week-old NOD mice. Claudin-5 level was increased and relocalized from the apical portion of neighboring endothelial cells to lateral membranes and cytoplasm in 12-week-old NOD mice. Additionally, the alteration of claudin-5 expression and distribution was further confirmed in labial salivary glands and bilateral parotid glands from SS patients. In cultured human microvessel endothelial cell line (HMEC-1), IFN-γ stimulation significantly increased claudin-5 expression. Taken together, we identified that the endothelial TJ barrier was disrupted and contributed to the development of salivary hyposecretion and lymphocytic infiltration in SS.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Endothelial tight junction barrier is disrupted in hyposecretory submandibular glands from Sjögren's syndrome mouse model </LI> <LI> The disrupted salivary endothelial barrier is linked with lymphocytic infiltration in Sjögren's syndrome mouse model </LI> <LI> The redistribution of claudin-5 is responsible for disrupted endothelial barrier in salivary glands from Sjögren's syndrome </LI> </UL> </P>

Ultrastructural changes of Haematococcus pluvialis (Chlorophyta) in process of astaxanthin accumulation and cell damage under condition of high light with acetate

He, Bangxiang,Hou, Lulu,Zhang, Feng,Cong, Xiaomei,Wang, Zhendong,Guo, Yalin,Shi, Jiawei,Jiang, Ming,Zhang, Xuecheng,Zang, Xiaonan The Korean Society of Phycology 2020 ALGAE Vol.35 No.3

Haematococcus pluvialis is a commercial microalga that can produce high quantities of astaxanthin. Under induced conditions, some important changes in the subcellular structures related to astaxanthin accumulation were observable. For example, a large number of astaxanthin granules, oil structures and starch granules appeared in the thick-walled cells; Astaxanthin granules gradually dissolved into the oil structures and spread throughout the entire cell with the fusion and diffusion process of oil structures during the middle and late stages of induction; The plastoglobules were closed to the newly formed structures, and some plastoglobules would abnormally increase in size under stress. Based on observations of cell damage, the degradation of membrane structures, such as chloroplasts, was found to be the primary form of damage during the early stage of induction. During the middle stage of induction, some transparent holes were exposed in the dissolving astaxanthin granules in the cytoplasm. In thick-walled cells, these transparent holes were covered by oil substances dissolving astaxanthin, thereby avoiding further damage to cells. Given the relatively few oil structures, in non-thick-walled cells, the transparent holes expanded to form multiple transparent areas, eventually resulting in the rupture and death of cells. These results suggested that the high level of synthesis and the wide range diffusion of oil explained the expansion of astaxanthin in H. pluvialis.

A Comparative Study of Unsupervised Deep Learning Methods for MRI Reconstruction

Zhuonan He,Cong Quan,Siyuan Wang,Yuanzheng Zhu,Minghui Zhang,Yanjie Zhu,Qiegen Liu 대한자기공명의과학회 2020 Investigative Magnetic Resonance Imaging Vol.24 No.4

Recently, unsupervised deep learning methods have shown great potential in image processing. Compared with a large-amount demand for paired training data of supervised methods with a specific task, unsupervised methods can learn a universal and explicit prior information on data distribution and integrate it into the reconstruction process. Therefore, it can be used in various image reconstruction environments without showing degraded performance. The importance of unsupervised learning in MRI reconstruction appears to be growing. Nevertheless, the establishment of prior formulation in unsupervised deep learning varies a lot depending on mathematical approximation and network architectures. In this work, we summarized basic concepts of unsupervised deep learning comprehensively and compared performances of several state-of-the-art unsupervised learning methods for MRI reconstruction.

An electrostatic discharge based needle-to-needle booster for dramatic performance enhancement of triboelectric nanogenerators

Zhai, Cong,Chou, Xiujian,He, Jian,Song, Linlin,Zhang, Zengxing,Wen, Tao,Tian, Zhumei,Chen, Xi,Zhang, Wendong,Niu, Zhichuan,Xue, Chenyang Elsevier 2018 APPLIED ENERGY Vol.231 No.-

<P><B>Abstract</B></P> <P>There is plenty of exploitable energy in the ambient environments. Triboelectric nanogenerator is an innovative electricity generation technology to convert the wasted mechanical energy into electrical energy. However, the output of conventional triboelectric nanogenerators cannot be employed efficiently because their tremendous internal resistance limits the current of electrons. Inspired by the principle of lightning rods, for the first time we propose the utilization of electrostatic discharge to improve the performance of triboelectric nanogenerators, which is realized by an opposite needles structure enclosed in an inert atmosphere. When this structure is connected in series with a vertical contact-separation triboelectric nanogenerator, the strong electric field near tips of two needles ionizes the gas around them, forming a conductive plasma channel between the tips, and releasing a mass of free charges. As a result, some exciting performances are obtained in triboelectric nanogenerator. The output peak-to-peak voltage is increased from 300 V to 1300 V, and the equivalent impedance of energy harvesting circuit is reduced from 100 MΩ to 10 kΩ. Especially in the optimal conditions, the maximum continuous power can even be significantly improved by 330.76%. Therefore, this work provides a new strategy for the energy conversion technology, which is significant for the advancement and application of triboelectric nanogenerators.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A strategy is proposed to improve the performance of triboelectric nanogenerators. </LI> <LI> The maximum continuous power can be boosted dramatically by electrostatic discharge. </LI> <LI> This design reduces the optimal impedance that is important for circuit matching. </LI> <LI> With this design triboelectric nanogenerators can directly drive low-power devices. </LI> <LI> This work is also significant for macro-energy conversion, such as ocean energy. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Low dose Emodin induces tumor senescence for boosting breast cancer chemotherapy via silencing NRARP

Zu, Cong,Qin, Guangyuan,Yang, Chunshu,Liu, Ning,He, Anning,Zhang, Mingdi,Zheng, Xinyu Elsevier 2018 Biochemical and biophysical research communication Vol.505 No.4

<P><B>Abstract</B></P> <P><B>Purpose</B></P> <P>The resistance to 5-FU often limits its clinical effectiveness on breast cancer treatment. Combination therapy thus is employed to overcome this treatment resistance. We here report a potent antitumor effect of Emodin at low dose on chemotherapy sensitivity of MCF-7 breast cancer cells.</P> <P><B>Methods</B></P> <P>Cell viability, apoptosis, glutathiones (GSH) concentration and Reactive oxygen species (ROS) activity following Emodin and 5-FU treatment was assessed. Cellular senescence following combined treatment and silence of NRARP was examined by senescence-associated β-galactosidase analysis. Western blot analysis was used to determine changes in the expression of p21, p16, p27, E2F1 and NRARP.</P> <P><B>Results</B></P> <P>Low dose Emodin potentiates 5-FU-induced apoptosis of breast cancer cells, in association with inhibition of NRARP, resulting in cellular senescence. RNA interference of NRARP induced cellular senescence in MCF-7 breast cancer cells. Furthermore, the cellular senescence induced by Emodin and 5-FU treatment could be reverted by pcDNA–NRARP.</P> <P><B>Conclusion</B></P> <P>These findings provide preclinical evidence for repurposing use of Emodin in combination with chemotherapeutic agents to treat breast cancer as an alternative salvage regimen.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The chemoresistance often limits clinical effectiveness of chemotherapy on breast cancer treatment. </LI> <LI> Emodin potentiates 5-FU-induced apoptosis of breast cancer cells. </LI> <LI> Low dose Emodin induces cellular senescence in breast cancer cells by inhibition of Nrarp. </LI> </UL> </P>

Elimination of moving vehicles effects on modal identification of beam type bridges

Wen-Yu He,Xu-Cong Ding,Wei-Xin Ren,Yue-Ling Jing 국제구조공학회 2021 Smart Structures and Systems, An International Jou Vol.28 No.3

The modal parameters identified under operation conditions are normally employed for bridge damage detection. However, the moving vehicles are usually deemed as part of the operation conditions without considering their mass property. Thus, the identified modal parameters belong to the vehicle-bridge system rather than the bridge itself, which would affect the effectiveness of subsequent damage detection. In this paper, the effects of moving vehicles on the identified frequencies and mode shapes under operation conditions are investigated via finite element model. The necessary of considering the moving vehicle effects is demonstrated by comparing the modal parameters variations induced by the moving vehicle and bridge damage. Then the empirical formulas to eliminate the moving vehicle effects considering the vehicle mass, velocity, bridge span and relative position are established by using the orthogonal test and least square method. Finally, examples are conducted to verify of the effectiveness of the proposed empirical formulas.

Comparison of Alpha-Factor Preprosequence and a Classical Mammalian Signal Peptide for Secretion of Recombinant Xylanase xynB from Yeast Pichia pastoris

( Zu Yong He ),( Yuankai Huang ),( Yufeng Qin ),( Zhiguo Liu ),( Delin Mo ),( Peiqing Cong ),( Yaosheng Chen ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.4

The secretory efficiency of recombinant xylanase xynB from yeast Pichia pastoris between the α-factor preprosequence and a classical mammalian signal peptide derived from bovine β-casein was compared. The results showed that although the bovine β-casein signal peptide could direct highlevel secretion of recombinant xylanase, it was relatively less efficient than the α-factor preprosequence. In contrast, the bovine β-casein signal peptide caused remarkably more recombinant xylanase trapped intracellularly. Realtime RT-PCR analysis indicated that the difference in the secretory level between the two signal sequences was not due to the difference in the transcriptional efficiency.