Development of High-specificity Antibodies against Renal Urate Transporters Using Genetic Immunization

( Guo Shuang Xu ),( Xiang Mei Chen ),( Di Wu ),( Suo Zhu Shi ),( Jian Zhong Wang ),( Rui Ding ),( Quan Hong ),( Zhe Feng ),( Shu Peng Lin ),( Yang Lu ) 생화학분자생물학회 2006 BMB Reports Vol.39 No.6

Screening and Anti-virulent Study of N-Acyl Homoserine Lactones DNA Aptamers against Pseudomonas aeruginosa Quorum Sensing

Zu-Guo Zhao,Yun Mei Yu,Bi Yu Xu,Shuang-Shuang Yan,Jun-Fa Xu,Fang Liu,Guo-Ming Li,Yuan Lin Ding,Shu Qing Wu 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2

In Pseudomonas aeruginosa, a quorum sensing (QS) system regulates the expression of many virulence factors. N-acyl homoserine lactone (HSL) is the signal molecule of QS system. In order to find a novel HSL binder to interfere with QS signaling and to attenuate P. aeruginosa virulence, an amino lactam surrogate (ALS) of HSL was used as a target to screen HSL aptamers with the technique of systematic evolution of ligands by exponential enrichment (SELEX). Eight HSL aptamers with high affinities for 3O-C12-HSL (20 nM ≤ Kd < 35 nM) or C4-HSL (25 nM < Kd < 50 nM) were finally obtained. In vitro QS-inhibiting study of P. aeruginosa showed that HSL aptamers could inhibit virulence in a dose-dependent manner. ALSap-8 which bound C4-HSL primarily acted on the rhl system and inhibited the secretion of pyocyanin. ALSap-5 which bound 3O-C12-HSL not only showed strong inhibitory activity on biofilm formation as well as secretions of LasA protease and LasB elastase, but also reduced pyocyanin secretion. Since the las system is capable of activating the rhl system mildly, we speculated that ALSap-5 can simultaneously interfere with the las and rhl systems. High-affinity aptamers against HSL in this study are novel QS and virulence-inhibitors, and may have potential as drug candidates for the treatment of P. aeruginosa infection.

An Aqueous Extract of Yunnan Baiyao Inhibits the Quorum-Sensing-Related Virulence of Pseudomonas aeruginosa

Zu-Guo Zhao,Shuang-Shuang Yan,Yun-Mei Yu,Na Mi,La-Xi Zhang,Jun Liu,Xiao-Ling Li,Fang Liu,Jun-Fa Xu,Wei-Qing Yang,Guo-Ming Li 한국미생물학회 2013 The journal of microbiology Vol.51 No.2

Yunnan Baiyao is a famous Chinese medicine that has long been directly applied to wounds to reduce bleeding, pain, and swelling without causing infection. However, little is known about its ability to prevent infection. The present study aimed to assess in vitro the anti-virulence activity of an aqueous extract of Yunnan Baiyao (YBX) using Pseudomonas aeruginosa as a pathogenic model. We found that a sub-MIC (2.5 mg/ml) of YBX can efficiently interfere with the quorum-sensing (QS) signaling circuit. Real-time polymerase chain reaction analysis showed that a sub-MIC of YBX downregulated the transcriptions of lasR, lasI, rhlR, and rhlI,which resulted in global attenuation of QS-regulated virulence activities, such as biofilm formation, and secretion of LasA protease, LasB elastase and pyocyanin. Further, YBX reduced the motility of P. aeruginosa related to QS, and impaired the formation of biofilms. These results suggest that YBX may possess global inhibitory activity against the virulence of P. aeruginosa and that YBX may also exhibit antimicrobial activity in vivo. The present study suggests that Yunnan Baiyao represents a potential source for isolating novel, safe, and efficacious antimicrobial agents.

An Increase in Synaptic NMDA Receptors in the Insular Cortex Contributes to Neuropathic Pain

Qiu, Shuang,Chen, Tao,Koga, Kohei,Guo, Yan-yan,Xu, Hui,Song, Qian,Wang, Jie-jie,Descalzi, Giannina,Kaang, Bong-Kiun,Luo, Jian-hong,Zhuo, Min,Zhao, Ming-gao AAAS 2013 Science signaling Vol.6 No.275

<P><B>Stopping the Pain</B></P><P>Damage to the central or peripheral nervous system can trigger the development of neuropathic pain, which can manifest as painful sensations in response to stimuli that are not normally painful. Qiu <I>et al.</I> found that mice that had developed neuropathic pain after peripheral nerve injury showed changes in synaptic plasticity and increased abundance of synaptic NMDA receptors in the insular cortex, a region of the brain that is activated by acute and chronic pain. Using pharmacological inhibitors and transgenic mice, they mimicked these changes in vitro with insular cortical slices and thus identified the signaling pathway responsible. Mice injected with NMDA receptor inhibitors showed reduced behavioral signs of neuropathic pain after peripheral nerve injury. Thus, blocking NMDA receptor function in the insular cortex may prevent the development of neuropathic pain.</P>

Study on the Joining of 2D Microstructure during the Fabrication of 3D Micro-Mold

Bin Xu,Xiao-yu Wu,Jian-guo Lei,Feng Luo,Chen-lin Du,Shuang-chen Ruan,Zhen-long Wang 한국정밀공학회 2014 International Journal of Precision Engineering and Vol. No.

The 3D micro-mold which is fabricated by the femtosecond laser cutting and micro electric resistance slip welding is formed throughthe lamination of multilayer 2D microstructures. By using this technology, the 3D micro-mold with a high depth-width ratio can bemanufactured. In order to improve the laminated precision and joining strength of each layer of 2D microstructure, this researchapplied layer-by-layer micro electric resistance slip welding to weld each layer of 2D microstructure. Firstly, the proper technicalparameters were obtained through the experiments of layer-by-layer micro electric resistance slip welding. Secondly, through thelayer-by-layer micro electric resistance slip welding, multilayer 2D microstructures were weld together and a 3D micro-mold wasformed. Moreover, the anti-shear stress test of 3D micro-mold was done. In the layer-by-layer slip welding process, electrode couldproduce some losses and the losses could deposit on the surface of micro-mold. This research also studied the deposition effect ofthe electrode. Finally, based on the above studies, the micro square-hole array and the micro-gear cavity were fabricated by joiningmultilayer 2D microstructure.

Molecular characterization and functional analysis of two trehalose transporter genes in the cabbage beetle, Colaphellus bowringi

Li Jia-Xu,Cao Zhen,Guo Shuang,Tian Zhong,Liu Wen,Zhu Fen,Wang Xiao-Ping 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.3

The trehalose, major blood sugar in insects, enhances stress resistance of diapausing individuals in adverse environment and provides an energy source for reproduction. Trehalose transporters (TRETs) play an important role in transport of trehalose from trehalose-producing tissues, e.g. fat body, to trehalose-consuming tissues. Although studies have shown that trehalose contributes to diapause and reproduction, the function of TRETs in these processes remains unclear. In this work, we cloned two TRET genes, TRET1a and TRET1b, from the cabbage beetle Colaphellus bowringi, which is capable of entering reproductive diapause under long-day conditions. We also analyzed the expression profiles of these two genes and investigated their potential roles in diapause and reproduction. The results suggested that both TRET1a and TRET1b belong to sugar-transporter and major facilitator superfamilies. Interestingly, TRET1a was highly expressed in the fat bodies of diapause-destined (DD) females but TRET1b was predominantly expressed in the ovaries of non-diapause-destined (NDD) females. Hormonal induction indicated that juvenile hormone induced TRET1b but repressed TRET1a at transcriptional levels. Methoprene-tolerant and Krüppel homolog 1 mediated the JH-suppressed TRET1a expression but were not involved in the regulation of TRET1b expression by JH. RNAi of TRET1a in DD females elevated the trehalose content in the fat bodies and suppressed the expression of a couple of genes related to stress resistance, which is a critical diapause trait. Knockdown of TRET1b in NDD females reduced the trehalose content in the ovaries but had no apparent effect on the ovary development and yolk deposition. These data suggest that TRET1a and TRET1b could regulate the trehalose content in specific tissues and may play potential roles in reproductive diapause in the females of C. bowringi. Introduction Diapause is a survival strategy for many insects in adapting to adverse environmental conditions (Tauber and Tauber, 1976; Tougeron, 2019). Diapause is a dynamic successive process that consists of diapause induction, preparation, initiation, maintenance, termination, and post-diapause quiescence (Kostal, 2006). Insects accumulate enormous nutrients such as sugars and lipids to enter diapause during diapause preparation phase (Hahn and Denlinger, 2007, 2011). The trehalose is the major hemolymph sugar in insects. The sugar not only provides source of energy but also protects proteins and cellular membranes from dehydration, desiccation, heat, cold, and oxidation (Elbein et al., 2003; Li et al., 2002; Richards et al., 2002; Wyatt and Kalf, 1957; Yancey, 2005; Zhu et al., 2008). Therefore, trehalose can play roles in reproduction and diapause by mediating energy supply and stress tolerance, respectively (Kamei et al., 2011; Lu et al., 2019; Xu et al., 2009).

The Beneficial Effects of Electroacupuncture at PC6 Acupoints (Neiguan) on Myocardial Ischemia in ASIC3 / mice

Ying-Wang,Yi-guo Chen,Wan-shuang Zhao,Di Li,Ya-han Xu,Meng-di Li,Jin Chen,Zhi-jun Kou,Qi-ge Wang,Nsoa dimitri Joseph 사단법인약침학회 2018 Journal of Acupuncture & Meridian Studies Vol.11 No.3

This study aims to investigate the possible mechanisms of electroacupuncture (EA) at PC6to improve myocardial ischemia (MI) by regulating the cardiac transient outward potassiumcurrent channel (Ito). According to the random number table, the mice were dividedinto six groups of six mice each: control group, MI group, PC6, LU7 (Lieque-point), ST36(Zusanli-point), and nonacupoint group. Mice in the control group were injected with saline(20 mg/kg, 24 hours interval), and the other ASIC3 / mice were injected subcutaneouslytwice with isoproterenol (ISO) (20 mg/kg, 24 hours interval). In thepreexperiment, 5 mg/kg, 10 mg/kg, 20 mg/kg, and 30 mg/kg of ISO were used, andthe results showed that 5 mg/kg and 10 mg/kg of ISO both could induce acute MI, butshorter duration of sustained MI. On the other hand, an injection of 30 mg/kg can makethe mice experience arrhythmia or die immediately, and EA was operated at PC6, LU7,ST36 acupoints, and nonacupoint in the mice of PC6, LU7, ST36, and nonacupoint groups,respectively, after injecting twice. Then Western blotting techniques (Western Blot) wereused to analyze the protein expressions of Kv1.4, Kv4.2, Kv4.3, and KchIP2. The results ofthis experiment showed that the protein expressions of Kv1.4, Kv4.2, Kv4.3, and KChIP2 inMI group were significantly lower than those in the control group (p < 0.01). Comparedwith MI group, the results of PC6, LU7, and ST36 groups obviously increased (p < 0.05). Furthermore, the expressions of PC6 group were higher than LU7 group and ST36 group(p < 0.05). And electrocardiogram’s T-waves showed obvious pathological changes inthe MI group compared to the control group (p < 0.01). After EA, the abnormal T-waves voltage of ECG in PC6, LU7, and ST36 groups was improved (p < 0.05). In addition, therate change of PC6 group was larger than that of both LU7 and ST36 groups (p < 0.05). But the T-waves voltage of the nonacupoint group was not significantly different than thatof the MI group (p > 0.05).

Protocatechuic Aldehyde Represses Proliferation and Migration of Breast Cancer Cells through Targeting C-terminal Binding Protein 1

Yu Deng,Wanjun Guo,Guancheng Li,Shuang Li,Hong Li,Xinyan Li,Bei Niu,Mingzhu Song,Yamei Zhang,Zhijian Xu,Fulun Li 한국유방암학회 2020 Journal of breast cancer Vol.23 No.1

Purpose: C-terminal binding protein 1 (CtBP1) is a transcriptional co-repressor that is overexpressed in many cancers. CtBP1 transcriptionally represses a broad array of tumor suppressors, which promotes cancer cell proliferation, migration, invasion, and resistance to apoptosis. Recent studies have demonstrated that CtBP1 is a potential target for cancer therapy. This study was designed to screen for compounds that potentially target CtBP1. Methods: Using a structure-based virtual screening for CtBP1 inhibitors, we found protocatechuic aldehyde (PA), a natural compound found in the root of a traditional Chinese herb, Salvia miltiorrhiza, that directly binds to CtBP1. Microscale thermophoresis assay was performed to determine whether PA and CtBP1 directly bind to each other. Further, clustered regularly interspaced short palindromic repeats associated Cas9 nuclease-mediated CtBP1 knockout in breast cancer cells was used to validate the CtBP1 targeting specificity of PA. Results: Functional studies showed that PA repressed the proliferation and migration of breast cancer cells. Furthermore, PA elevated the expression of the downstream targets of CtBP1, p21 and E-cadherin, and decreased CtBP1 binding affinity for the promoter regions of p21 and E-cadherin in breast cancer cells. However, PA did not affect the expression of p21 and E-cadherin in the CtBP1 knockout breast cancer cells. In addition, the CtBP1 knockout breast cancer cells showed resistance to PA-induced repression of proliferation and migration. Conclusion: Our findings demonstrated that PA directly bound to CtBP1 and inhibited the growth and migration of breast cancer cells through CtBP1 inhibition. Structural modifications of PA are further required to enhance its binding affinity and selectivity for CtBP1.

Dynamic changes of multi-notoginseng stem-leaf ginsenosides in reaction with ginsenosidase type-I

Yongkun Xiao,Chun-Ying Liu,임완택,Shuang Chen,Kangze Zuo,Hong Shan Yu,Jian-Guo Song,Long-Quan Xu,Tea-Hoo Yi,Feng Xie Jin 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.2

Background: Notoginseng stem-leaf (NGL) ginsenosides have not been well used. To improve their utilization, the biotransformation of NGL ginsenosides was studied using ginsenosidase type-I from Aspergillus niger g.848. Methods: NGL ginsenosides were reacted with a crude enzyme in the RAT-5D bioreactor, and the dynamic changes of multi-ginsenosides of NGL were recognized by HPLC. The reaction products were separated using a silica gel column and identified by HPLC and NMR. Results: All the NGL ginsenosides are protopanaxadiol-type ginsenosides; the main ginsenoside contents are 27.1% Rb3, 15.7% C-Mx1, 13.8% Rc, 11.1% Fc, 7.10% Fa, 6.44% C-Mc, 5.08% Rb2, and 4.31% Rb1. In the reaction of NGL ginsenosides with crude enzyme, the main reaction of Rb3 and C-Mx1 occurred through Rb3/C-Mx1/C-Mx; when reacted for 1 h, Rb3 decreased from 27.1% to 9.82 %, C-Mx1 increased from 15.5% to 32.3%, C-Mx was produced to 6.46%, finally into C-Mx and a small amount of C-K. When reacted for 1.5 h, all the Rb1, Rd, and Gyp17 were completely reacted, and the reaction intermediate F2 was produced to 8.25%, finally into C-K. The main reaction of Rc (13.8%) occurred through Rc/C-Mc1/CMc/ C-K. The enzyme barely hydrolyzed the terminal xyloside on 3-Oe or 20-O-sugar-moiety of the substrate; therefore, 9.43 g C-Mx, 6.85 g C-K, 4.50 g R7, and 4.71 g Fc (hardly separating from the substrate) were obtained from 50 g NGL ginsenosides by the crude enzyme reaction. Conclusion: Four monomer ginsenosides were successfully produced and separated from NGL ginsenosides by the enzyme reaction.

Metal organic frameworks template-directed fabrication of rod-like hollow BiOClxBr1x with adjustable band gap for excellent photocatalytic activity under visible light

Ze Luo,Jinlong Li,Guozhe Sui,Yan Zhuang,Dongxuan Guo,Rongping Xu,Shuang Liang,Hong Yao,Chao Wang,Shijie Chen 한국화학공학회 2022 Korean Journal of Chemical Engineering Vol.39 No.8

Developing an efficient, environmentally friendly, and pollution-free catalyst with excellent visible light catalyticactivity is a promising strategy for dye wastewater treatment. Herein, the rod-like hollow BiOClxBr1x (x=1, 0.75, 0.5,0.25, 0), with an adjustable band gap, was successfully prepared using Bi-based metal-organic framework as template. The corresponding hollow assembly and introduction of Br imparted valuable structural advantages and intrinsic characteristicsfor improved photocatalytic activity. Significantly, the degradation efficiency of BiOCl0.5Br0.5 for the RhodamineB (RhB) solution reached 92% under visible light illumination for 90 min, which is considerably higher than that ofCAU-17-derived Bi2O3 and BiOCl. Overall, these findings shed fundamental insight on constructing novel photocatalystswith excellent visible light driven photocatalytic activity and offered a new method for treating dye wastewater.