Development of a novel peptide microarray for large-scale epitope mapping of food allergens

Lin, Jing,Bardina, Ludmilla,Shreffler, Wayne G.,Andreae, Doerthe A.,Ge, Yongchao,Wang, Julie,Bruni, Francesca M.,Fu, Zhiyan,Han, Youngshin,Sampson, Hugh A. Elsevier 2009 The Journal of allergy and clinical immunology Vol.124 No.2

<P><B>Background</B></P><P>The peptide microarray is a novel assay that facilitates high-throughput screening of peptides with a small quantity of sample.</P><P><B>Objective</B></P><P>We sought to use overlapping peptides of milk allergenic proteins as a model system to establish a reliable and sensitive peptide microarray-based immunoassay for large-scale epitope mapping of food allergens.</P><P><B>Methods</B></P><P>A milk peptide microarray was developed by using commercially synthesized peptides (20-mers, 3 offset) covering the primary sequences of α<SUB>s1</SUB>-casein, α<SUB>s2</SUB>-casein, β-casein, κ-casein, and β-lactoglobulin. Conditions for printing and immunolabeling were optimized using a serum pool of 5 patients with milk allergy. Reproducibility of the milk peptide microarray was evaluated using replicate arrays immunolabeled with the serum pool, whereas specificity and sensitivity were assessed by using serial dilution of the serum pool and a peptide inhibition assay.</P><P><B>Results</B></P><P>Our results show that epitopes identified by the peptide microarray were mostly consistent with those identified previously by SPOT membrane technology, but with specific binding to a few newly identified epitopes of milk allergens. Data from replicate arrays were reproducible (<I>r</I> ≥ 0.92) regardless of printing lots, immunolabeling, and serum pool batches. Using the serially diluted serum pool, we confirmed that IgE antibody binding detected in the array was specific. Peptide inhibition of IgE binding to the same peptide and overlapping peptides further confirmed the specificity of the array.</P><P><B>Conclusion</B></P><P>A reliable peptide microarray was established for large-scale IgE epitope mapping of milk allergens, and this robust technology could be applied for epitope mapping of other food allergens.</P>

The Numerical Simulation Analysis of Flow Field in Level Control Valve of Water Storage Tank

Yaping Wang,Lei Gao,Jianghua Ge,Han Gao,Jing Zhang 보안공학연구지원센터 2014 International Journal of Control and Automation Vol.7 No.10

Level control valve of water storage tank is installed in the outlet of tank, which control the water level of water storage tank by adjusting the water emissions, which make great significance for the safe operation of the super (super) criticality thermal power unit boiler . Complete the simulation analysis of internal flow field by the method of numerical simulation ,in order to achieve structure improvements of level control valve of water storage tank, that make level control valve of water storage tank better meet the work requirements. Establish flow channel model and mesh models of level control valve of water storage tank in various stages of the open process. Make three dimensional viscous numerical simulation of the internal complex flow field by the computational fluid dynamics software of ANSYS CFX. The error between the simulation values of the flow coefficient and the theoretical values , trial values is less than 5% ,which verifies the correctness of the simulation results . Get pressure contour , velocity streamline diagrams and outlet mass flow of level control valve of water storage tank in various stages of the open process. According to the pressure contours and velocity streamline diagrams, make detailed analysis of the flow field of level control valve of water storage tank. According to the dates of outlet mass flow, fit the flow characteristic curve of level control valve of water storage tank. Provide a theoretical basis for the structural design of level control valve of water storage tank.

Novel Mutation of the NCSTN Gene Identified in a Chinese Acne Inversa Family

( Jing Wu ),( Huiyao Ge ),( Yiming Fan ),( Qi Zhen ),( Lili Tang ),( Liangdan Sun ) 대한피부과학회 2020 Annals of Dermatology Vol.32 No.3

Acne inversa is a chronic inflammatory follicular disease with autosomal dominant inheritance. In recent years, many functional mutations in the NCSTN genes have been identified as the cause of familial acne inversa. Herein, we recruited four patients and seven unaffected individuals from a Chinese family and performed Sanger sequencing of the NCSTN gene. One novel frameshift mutation, c.450_459del (p.Ser 151GlnfsX48), was identified in exon 5 of the NCSTN gene. Three normal-looking children carrying the mutation were proven to be patients. We also presented a literature review from previous studies of acne inversa, suggesting that NCSTN is a hotspot gene for acne inversa. Most affected individuals experienced onset in adolescence. We confirmed the diagnosis in this family based on the mutation. This finding will help expound the relationship between the NCSTN gene and the pathogenesis of acne inversa and emphasize the value of genetic diagnosis in monogenic disorder. (Ann Dermatol 32(3) 237∼242, 2020)

Predictive Value of Molecular Subtyping for Locoregional Recurrence in Early-Stage Breast Cancer with N1 without Postmastectomy Radiotherapy

Ge Wen,Jin-Shan Zhang,Yu-Jing Zhang,Yu-Jia Zhu,Xiao-Bo Huang,Xunxing Guan 한국유방암학회 2016 Journal of breast cancer Vol.19 No.2

Purpose: This study was designed to investigate the relationship between molecular subtype and locoregional recurrence (LRR) in patients with early-stage breast cancer with 1–3 positive axillary lymph nodes (ALNs) and improve the individualized indications for postmastectomy radiotherapy (PMRT). Methods: The records of 701 patients with pT1-2N1M0 breast cancer who did not undergo PMRT were retrospectively analyzed. Tumors were subclassified as follows: luminal A, luminal B, human epidermal growth factor receptor 2 (HER2)-enriched, and basal-like subtypes. Multivariate Cox analysis was used to determine the risk of LRR associated with the different subtypes and to adjust for clinicopathologic factors. Results: Luminal A, luminal B, HER2- enriched, and basal-like subtypes accounted for 51.2%, 28.0%, 8.1%, and 12.7% of cases, respectively. The median follow-up duration was 67 months (range, 9–156 months). Univariate analysis revealed that, compared with the luminal A subtype, the HER2-enriched and basal-like subtypes were associated with significantly higher 5-year LRR rates (5.6% vs. 21.6% and vs.15.7% respectively; p=0.002 each), lower 5-year LRR-free survival (LRFS) rates (90.6% vs. 73.8% and 78.5%, respectively; p=0.001 each), and poorer 5-year breast cancer-specific survival (BCSS) rates (93.7% vs. 82.2% [p=0.002] and 84.9% [p=0.001], respectively). Multivariate analysis revealed that the HER2-enriched and basal-like subtypes, age ≤35 years, a medial tumor, and pT2 stage were poor prognostic factors for LRR and LRFS; furthermore, 2 to 3 positive ALNs represented an independent prognostic factor affecting LRR. The 10-year LRR rates of patients with 0, 1, 2, 3, and 4 risk factors were 1.0%, 6.9%, 14.3%, 30.4%, and 54.3%, respectively (p<0.001); the 10-year BCSS rates were 86.6%, 88.5%, 84.4%, 79.7%, and 38.8%, respectively (p<0.001). Conclusion: Molecular subtyping allows for individualized evaluation of LRR risk in patients with pT1-2N1M0 breast cancer. PMRT should be recommended for patients with ≥3 LRR risk factors.

Vagococcus zengguangii sp. nov., isolated from yak faeces

Ge Yajun,Jin Dong,Lai Xin-He,Yang Jing,Lu Shan,Huang Ying,Zheng Han,Zhang Xiaoyan,Xu Jianguo 한국미생물학회 2021 The journal of microbiology Vol.59 No.1

Two unknown Gram-stain-positive, catalase- and oxidasenegative, non-motile, and coccus-shaped bacteria, designated MN-17T and MN-09, were isolated from yaks faeces (Bos grunniens) in the Qinghai-Tibet Plateau of China. 16S rRNA gene sequence-based comparative analyses revealed that the two strains were grouped within the genus Vagococcus, displaying the highest similarity with Vagococcus xieshaowenii CGMCC 1.16436T (98.6%) and Vagococcus elongatus CCUG 51432T (96.4%). Both strains grew optimally at 37°C and pH 7.0 in the presence of 0.5% (w/v) NaCl. The complete genome of MN-17T comprises 2,085 putative genes with a total of 2,190,262 bp and an average G + C content of 36.7 mol%. The major fatty acids were C16:0 (31.2%), C14:0 (28.5%), and C18:1ω9c (13.0%); the predominant respiratory quinone was MK-7 (68.8%); the peptidoglycan type was A4α(L-Lys-DAsp); and the major polar lipid was diphosphatidylglycerol. Together, these supported the affiliation of strain MN-17T to the genus Vagococcus. In silico DNA-DNA hybridization and the average nucleotide identity values between MN-17T and all recognized species in the genus were 21.6–26.1% and 70.7–83.0%, respectively. MN-17T produced acid from D-cellobiose, D-fructose, glycerol, D-glucose, N-acetyl-glucosamine, gentiobiose, D-mannose, D-maltose, D-ribose, Dsaccharose, salicin, D-trehalose, and D-xylose. These results distinguished MN-17T and MN-09 from closely related species in Vagococcus. Thus, we propose that strains MN-17T and MN-09 represent a novel species in the genus Vagococcus, with the name Vagococcus zengguangii sp. The type strain is MN-17T (= CGMCC 1.16726T = GDMCC 1.1589T = JCM 33478T).

Electroacupuncture Promotes Neural Proliferation in Hippocampus of Perimenopausal Depression Rats via Wnt/β-Catenin Signaling Pathway

Qin Jing,Lu Ren,Xue Deng,Nan Zhang,Martin Fu,Ge Wang,Xi-Rong Jiang,Shu-Ru Lin,Cai-Rong Ming 사단법인약침학회 2020 Journal of Acupuncture & Meridian Studies Vol.13 No.3

Background and Objective: Perimenopausal depression is caused by the impaired function of the ovarium before menopause and with a series of symptoms. Electroacupuncture (EA) therapy has been demonstrated to improve clinically depression. However, the mechanism underlying its therapeutic activity remains unknown. This study aimed to investigat the effects of EA treatment on the hippocampal neural proliferation through Wnt signaling pathway. Methods: Chronic unpredictable mild stress (CUMS) combined with bilateral ovariectomy (OVX) were used to establish a rat model of perimenopausal depression. The open field test (OFT) and sucrose preference test (SPT) were used to assess depression-like behaviors in rats. ELISAs were used to measure estrogen (E2), luteinizing hormone (LH) and gonadotropin-releasing hormone (GnRH) levels in the serum. RT-PCR and Western blot assay were utilized for measuring the mRNA expressions and protein expressions of GSK-3β/β-catenin. Results: Four-week EA treatment at three points including “Shenshu” (BL23), “Baihui” (GV20) and “Sanyinjiao” (SP6) simultaneously ameliorated depression-like behaviors in rats with CUMS and OVX, whereas rescued the decreased serum level of E2 and prevented the increased serum levels of GnRH and LH. EA treatment ameliorated CUMS and OVX-induced alterations of glycogen synthase kinase-3b (GSK-3β) and β-catenin mRNA levels, β-catenin and phosphorylated β-catenin (p-β-catenin) protein levels. Conclusions: The results showed that EA treatment promoted hippocampal neural proliferation in perimenopausal depression rats via activating the Wnt/b-catenin signaling pathway, indicating that EA may represent an efficacious therapy for perimenopausal depression.

[ $Ce^{4+}$ ]-Stimulated Ion Fluxes Are Responsible for Apoptosis and Taxol Biosynthesis in Suspension Cultures of Taxus Cells

Li Jing-Chuan,Ge Zhi-Qiang,Yuan Ying-Jin The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.2

Ion fluxes across the plasma membrane activated by 1 mM $Ce^{4+}$, cell apoptosis and taxol biosynthesis in suspension cultures of Taxus cusp/data were studied. The extracellular pH sharply decreased upon the addition of 1 mM $Ce^{4+}$, then increased gradually and exceeded the initial pH value over a time period of 12 h. The extracellular $Ca^{2+}$ concentration decreased within the first 3 h after the addition of $Ce^{4+}$, then gradually decreased to one third of initial value in control at about 72 h and remained unchanged afterwards. Experiments with an ion channel blocker and a $Ca^{2+}$-channel blocker indicated that the dynamic changes in extracellular pH and the $Ca^{2+}$ concentration resulted from the $Ce^{4+}$-induced activation of W uptake and $Ca^{2+}$ influx across the plasma membrane via ion channels. A pretreatment of the ion channel blocker initiated $Ce^{4+}$-treated cells to undergo necrosis, and the prior addition of the $Ca^{2+}$-channel blocker inhibited $Ce^{4+}$-induced taxol biosynthesis and apoptosis. It is thus inferred that W uptake is necessary for cells to survive a $Ce^{4+}$-caused acidic environment and is one of the mechanisms of $Ce^{4+}$-induced apoptosis. Furthermore, the $Ca^{2+}$ influx across the plasma membrane mediated both the $Ce^{4+}$-induced apoptosis and taxol biosynthesis.

Branched polymeric prodrug/programmed cell death 4 complexes for combinational cancer therapy

Yu‑Jing He,Lei Xing,Peng‑Fei Cui,Jia‑Liang Zhang,Jian‑Bin Qiao,Cheng‑Qiong Luo,Ge Jiang,Hu‑Lin Jiang 한국약제학회 2017 Journal of Pharmaceutical Investigation Vol.47 No.2

Here, we demonstrate a co-delivery system constructed by integrating chemotherapeutic molecules into branched polymeric prodrug which can condense nucleic acids. Demethylcantharidate (DCA) was chosen as a model drug and premodified through nucleophilic substitution reaction by its two carboxylic groups with allyl chloride. The synthesized intermediate (DCA-dially) was then used to polymerize with tris (2-aminoethyl) amine (TAEA) through progressively ammonolysis reaction. The obtained poly (DCA-alt-TAEA) (DCAT) was used to pack PDCD4 into spherical-like nanoparticles through electrostatic interaction. Gel retardation assays implied that DCAT could integrate DNA at the weight ratio of 1 and protect it from digestion by nuclease. Acid-base titration experiments showed that DCAT obtained preferable buffering capability which was beneficial for the endosomal escape of DCAT/PDCD4 complexes. Cellular tests involving gene transfection efficiency and cytotoxicity indicated that DCA and PDCD4 co-delivered by the complexes significantly and synergistically suppressed the viability of SMMC-7721 cells. These results suggest that integrating chemotherapeutic molecules into nucleic acid-packing polymeric prodrug as cationic polymer/PDCD4 complexes is a highly efficient way to co-deliver chemotherapeutic drugs and nucleic acids for cancer therapy.

COINCIDENCE THEOREMS FOR NONCOMPACT RC-MAPS IN ABSTRACT CONVEX SPACES WITH APPLICATIONS

Ming-ge Yang,Nan-Jing Huang 대한수학회 2012 대한수학회보 Vol.49 No.6

In this paper, a coincidence theorem for a compact RC-map is proved in an abstract convex space. Several more general coincidence theorems for noncompact RC-maps are derived in abstract convex spaces. Some examples are given to illustrate our coincidence theorems. As ap-plications, an alternative theorem concerning the existence of maximal elements, an alternative theorem concerning equilibrium problems and a minimax inequality for three functions are proved in abstract convex spaces. In this paper, a coincidence theorem for a compact RC-map is proved in an abstract convex space. Several more general coincidence theorems for noncompact RC-maps are derived in abstract convex spaces. Some examples are given to illustrate our coincidence theorems. As ap-plications, an alternative theorem concerning the existence of maximal elements, an alternative theorem concerning equilibrium problems and a minimax inequality for three functions are proved in abstract convex spaces.

Enhanced Nitrogen Photo Fixation Performance of Transition Metal-Doped Urchin-Like W18O49 Under Visible-Light Irradiation

Jianhua Ge,Jing Xu,Yujie Liu,Long Zhang,Lan Wang,Dehan Wu 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2019 NANO Vol.14 No.11

Nitrogen photo fixation mediated by various photocatalysts has received extensive concerns in recent years. In this work, a general strategy is proposed to enhance nitrogen photo fixation performance with some W18O49-based photocatalysts under visible-light irradiation. A series of photocatalysts were successfully synthesized in the presence of some noble-metal-free transition metals (Fe, Co and Ni) to modify the urchin-like W18O49. The resulting photocatalysts were systematically characterized using XRD, SEM, EDS, XPS, PL, N2 adsorption–desorption isotherms, and DRS. Structural characterization of the photocatalysts revealed that the crystal morphology features were preserved, optional harvesting properties were enhanced, BET specific surface area was extended, and migration efficiency of the photo-induced charge carriers was significantly improved in the modified-W18O49 materials. Therefore, a progressed nitrogen photo fixation performance was observed. The maximum production of NH4 + was approximately reached to 2752 μg · g -1 catal. under the optimum reaction conditions. Furthermore, a plausible nitrogen photo fixation mechanism in the presence of the synthesized composites is also proposed.