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Screening of the Antigen Epitopes of Basic Fibroblast Growth Factor by Phage Display
Xiang, Junjian,Zhong, Zhenyu,Deng, Ning,Zhong, Zhendong,Yang, Hongyu Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.3
In order to investigate the epitope of basic fibroblast growth factor (bFGF) and its immunogenicity, the epitopes of bFGF were screened from the phage display library with monoclonal antibody GF22, which can neutralize the bio-activity of bFGF. By three rounds of screening, the positive phage clones with bFGF epitopes were selected, which can effectively block the bFGF to bind with GF22. Sequence analysis showed that the epitopes shared a highly conservative sequence (Leu-Pro-Pro/Leu-Gly-His-Phe/Ile-Lys). The sequence of PPGHFK was located at 22-27 of the bFGF. The specific immuno-response of mouse could be highly induced by phage clones with the epitopes. And the anti-bFGF activity induced by LPGHFK was 3 times higher than the original sequence, which showed that the mimetic peptide LPLGHIK might be used as a tumor vaccine in the prevention and treatment of tumor.
Xiang Yaoxian,Dai Junxi,Li Yao,You Zongqi,Zhang Junpeng,Huang Xinying,Nie Shuqi,Chen Yujie,Xu Lei,Liu Fengming,Jiang Junjian,Xu Jianguang 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-
Neutrophils are the earliest master inflammatory regulator cells recruited to target tissues after direct infection or injury. Although inflammatory factors are present in muscle that has been indirectly disturbed by peripheral nerve injury, whether neutrophils are present and play a role in the associated inflammatory process remains unclear. Here, intravital imaging analysis using spinning-disk confocal intravital microscopy was employed to dynamically identify neutrophils in denervated muscle. Slice digital scanning and 3D-view reconstruction analyses demonstrated that neutrophils escape from vessels and migrate into denervated muscle tissue. Analyses using reactive oxygen species (ROS) inhibitors and flow cytometry demonstrated that enhanced ROS activate neutrophils after denervation. Transcriptome analysis revealed that the vast majority of neutrophils in denervated muscle were of the CXCR2 subtype and were recruited by CXCL1. Most of these cells gradually disappeared within 1 week via P53-mediated apoptosis. Experiments using specific blockers confirmed that neutrophils slow the process of denervated muscle atrophy. Collectively, these results indicate that activated neutrophils are recruited via chemotaxis to muscle tissue that has been indirectly damaged by denervation, where they function in delaying atrophy.
EID3 Promotes Glioma Cell Proliferation and Survival by Inactivating AMPKα1
Xiang, Yaoxian,Zhu, Lei,He, Zijian,Xu, Lei,Mao, Yuhang,Jiang, Junjian,Xu, Jianguang The Korean Neurosurgical Society 2022 Journal of Korean neurosurgical society Vol.65 No.6
Objective : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.
Production of Recombinant Humanized Anti-HBsAg Fab Fragment from Pichia pastoris by Fermentation
Deng, Ning,Xiang, Junjian,Zhang, Qing,Xiong, Sheng,Chen, Wenyin,Rao, Guirong,Wang, Xunzhang Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.3
In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.
Shaohua Chen,Xuan Liu,Shengyang Dai,Wenfeng Xiang,Haizhong Guo,Junjian Li 한국물리학회 2020 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol.76 No.2
CuxO (Cupric Oxide) nanotubes (NTs) were synthesized by using thermal oxidation of Cu nanowires (NWs) for 2 and 4 hours, respectively, and the effect of the components on their photoelectrical properties due to the infrared (IR) light with a wavelength of 1064 nm were investigated. In the 2-h CuxO NTs, the main component was Cu2O; however, the 4-h CuxO NTs were composed of a mixture of CuO and Cu2O. The photoelectric properties showed that the voltage difference and the sensitivity of 4-h CuxO NTs were more than 3 times those of the 2-h CuxO NTs, and its linear range was only about one-third that of the 2-h CuxO NTs. The above results were caused by the 4-h CuxO NTs containing a higher content of CuO, which had a suitable bandgap of 1.2 eV for light with a wavelength of 1064 nm. In addition, the response and the recovery times for the 4-h NTs were on the same order as those of the 2-h NTs. The conclusions from this article indicated that 4-h and 2-h NTs were very promising for the design of IR photodetector device with higher sensitivity and a larger detection range respectively.