1.Cloning and Expression of an Immunodominant Fragment of Glycoprotein G of HSV-2 Using Directional TOPO Vector in E.coli
Shuqi WANG ; Yueping YIN ; Yanhua YU ; Weihong LAI ; Mingjun JIANG ; Qiang CHEN ; Xiangsheng CHEN
Chinese Journal of Dermatology 1994;0(05):-
Objective To clone and express immunodominant fragment of glycoprotein G of HSV-2 (FgG-2). Methods The target gene was amplified by polymerase chain reaction (PCR). The PCR products were ligated into directional TOPO expression vector. After identification, the recombinant expression vector was transferred into BL21 StarTM cell for expression. Finally, recombinant protein of FgG-2 (rFgG-2) was detected by Western Blot (WB). Results A 616 bp DNA fragment was obtained with PCR and then confirmed in recombinant vector by PCR and sequencing, bearing 99.5% consistent sequence with target gene. Highest recombinant protein production was obtained at the time point of 3 hours. Expression of target protein was confirmed by WB with anti-gG monoclonal antibody. Conclusions The immunodominant fragment of gG-2 has been successfully cloned and expressed in E.coli, which might be used for the development of serum diagnostics assay kits for HSV-2 infection.
2.hARD1 antiserum preparation and primary immunohistochemical analysis of hARD1 in tumor tissues.
Min YU ; Chao HUANG ; Mingjun XIANG ; Jianhua LAI ; Hui YANG ; Mingxing MA ; Deyong TAN
Chinese Journal of Biotechnology 2008;24(7):1155-1161
Human arrest defective 1 (hARD1) is an acetyltransferase; its physiological significance remains unclear. To explore the relationship between ARD1 protein and tumors, we detected the hARD1 protein in tumor tissues in vivo. We cloned hARD1 gene from Hela cell and construct recombinant plasmid pET28b-hARD1. The recombinant plasmid was transformed into E. coli BL21 (DE3)plysS. hARD1 protein was expressed by inducing with IPTG(1 mmol/L) and purified up to 95% through Ni2+ chelation affinity chromatography. We used the purified hARD1 protein as antigen immunized the Balb/c mice and obtained the hARD1 specific polyclonal antiserum. Through immunohistochemical analysis of different tumor tissues in vivo, we found that hARD1 expressed at high frequency in breast cancer, prostate cancer and lung cancer, especially, hARD1 expression frequency in breast cancer was up to 70%, which is higher than in the other tumors. These results indicate that the high expression level of hARD1 could be an indicator of the breast cancer. This new finding would be a foundation to further explore the relationship between breast tumor and hARD1.
Acetyltransferases
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analysis
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genetics
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immunology
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Amino Acid Sequence
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Animals
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Antibodies
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blood
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immunology
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Base Sequence
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Biomarkers, Tumor
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Breast Neoplasms
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metabolism
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pathology
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Cell Line, Tumor
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Female
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Humans
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Immune Sera
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biosynthesis
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Immunization
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Immunohistochemistry
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Lung Neoplasms
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metabolism
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pathology
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Male
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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N-Terminal Acetyltransferase A
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N-Terminal Acetyltransferase E
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Prostatic Neoplasms
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metabolism
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pathology
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
4.Diagnostic value of sensory sparing patterns in childhood immune-mediated acute or chronic inflammatory polyneuropathy
Mingjun LAI ; Xiaoping PAN ; Changchun SU ; Haobo CHEN ; Qiusheng CHENG ; Ze LI
Chinese Journal of Neuromedicine 2019;18(10):1019-1024
Objective To investigate the characteristics and significance of sural sparing pattern and radial sparing pattern in children immune-mediated acute or chronic inflammatory peripheral neuropathy.Methods Forty children with Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) aged<14 years, admitted to our hospital from September 2014 to August 2019, were chosen as immune-inflammatory group; 15 children (<14 years old) with multiple peripheral neuropathy of other etiologies from the same source were classified as other etiologies group. The characteristics of sensory nerve damage measured by nerve conduction velocity in the two groups were compared. The existences of sural sparing (SS) pattern and radial sparing (RS) pattern, and sensory ratio (SR)>1 were conformed. SR=(sural nerve sensory nerve action potentials+radial nerve sensory nerve action potentials)/(median nerve sensory nerve action potentials+ulnar nerve sensory nerve action potentials).Results (1) In 40 patients from immune-inflammatory group, 38 were GBS and two were with CIDP; Among the 38 with GBS, 34 were with acute inflammatory demyelinatingpolyneuropathies (AIDP), 3 were with acute motor axonal neuropathy (AMAN), and one was with acute motor sensory axonal neuropathy. Among the 15 patients from the other etiologies group, 9 were with charcot-marie-tooth (CMT) type 1, one was with CMT type 2, one was with CMT intermediate type, and two were with hereditary neuropathy with liability to pressure palsies (HNPP), and two were with mitochondrial disease involves polyneuropathy. (2) As compared with patients from other etiologies group, patients from immune-inflammatory group had significantly higher positive rates of SS (72.5%vs. 6.7%) , RS(42.5%vs.6.7%) and SR>1 (75.0%vs.13.3%). In the diagnoses of childhood acute or chronic inflammatory polyneuropathy, the sensitivity of SS pattern was 0.73, specificity was 0.93, and positive likelihood ratio was 10.88. SR>1 had moderate specificity (0.87) and low sensitivity (0.41).Conclusion The sensory sparing patterns, especially SS pattern, have valuable clinical significance in the diagnoses of acute or chronic inflammatory peripheral neuropathy in children.
5.Construction of artificial intelligence cloud platform for multi-center digestive endoscopy in Shandong Province (with video)
Guangchao LI ; Zhen LI ; Yusha ZHAO ; Jing LIU ; Ruchen ZHOU ; Mingjun MA ; Xuejun SHAO ; Yonghang LAI ; Xiuli ZUO ; Yanqing LI
Chinese Journal of Digestion 2022;42(5):328-335
Objective:Based on the artificial intelligence (AI) technology in endoscopy and the internet platform, to explore and construct a safe, standardized, scientific and rigorous database for digestive endoscopy, and to provide reference and evidence for the data quality control of AI in digestive endoscopy in China.Methods:After referring to relevant guidelines and standards, data collection and labelling standards of digestive endoscopy of 12 common gastrointestinal diseases were determined. The software of online collection and labelling of multi-center digestive endoscopy data in Shandong Province was developed. Endoscopic equipment with a domestic market share of >5% was used and dozens of experienced endoscopists from 9 medical centers in Shandong Province were uniformly trained for data labelling. From July 2019 to July 2020, the endoscopic examination data from 9 medical centers including Qilu Hospital of Shandong University, Shandong Provincial Hospital , Liaocheng People′s Hospital, Linyi People′s Hospital, Weihai Municipal Hospital, Taian City Central Hospital, Binzhou Medical University Hospital, Yantai Yuhuangding Hospital and Qilu Hospital of Shandong University (Qingdao) were prospectively and continuously collected and labeled. The optimized, desensitized, and generalized data were uploaded to the server. After the file synchronization, data processing, and expert review, a multi-center digestive endoscopy AI database with standard data collection and labelling in Shandong Province was constructed, namely cloud platform. Descriptive methods were used for statistical analysis.Results:The collection and labelling standards for multi-center digestive endoscopy AI data in Shandong province was established. The software of online collection and labelling of multi-center digestive endoscopy AI data in Shandong province was developed. The database in Shandong province was successfully constructed. In the database, 43 010 lesions, 40 353 images, and 11 289 examinations were labeled. Among them, there were 2 906 cases of early esophageal cancer, 2 912 cases of early gastric cancer, 2 397 cases of early colorectal cancer, and 9 773 cases of colorectal polyps (5 539 cases of adenomatous polyps, 1 161 cases of non-adenomatous polyps and 3 073 case of undetermined polyps).Conclusions:The multi-center AI cloud platform for digestive endoscopy in Shandong Province adopts unified standards and collection and labeling software, which ensures the safety and standardization of endoscopy data. It provides a reference and basis for the construction of a quality control system for standardized data collection and labelling of digestive endoscopy AI data in our country and for the third-party data supervision.