1.Bellybutton exposure in premature nursing
Hequn WEN ; Lixing ZHU ; Qiaoping ZENG ; Jingfeng DU
Modern Clinical Nursing 2014;(6):70-72
Objective To explore the effect of bellybutton exposure on premature navel nursing effect.Method Three hundred and ninety-three premature infants were randomly divided into experiment group(n=195)and control group(198). Bellybutton fasciation and bellybutton exposure were used in the control group and the experiment group respectively. The two groups were compared in terms of umbilical shedding,time of umbilical healing,umbilical swelling,bleeding and breathing.Result The umbilical healing time in the experiment group was significantly shorter than that of the control group(P<0.05). The incidences of omphalitis,umbilical swelling and bleeding, changed breathing in the experiment group were significantly lower than those of the control group(all P<0.05).Conclusion The bellybutton exposure is effective in promoting the umbilical shedding in premature infants and reducing the incidence rate of premature navel infection.
2.Construction of survivin shRNA and APC double gene vector and its expression in HT-29 cells
Xixian YUAN ; Houyu DUAN ; Feng CAO ; Jingfeng DU ; Yanli ZHU ; Kai CHENG ; Yantao YANG
Journal of Medical Postgraduates 2016;29(4):369-374
Objective Studies show that the abnormal ex-pressions of APC and survivin play important roles in the development and progression of colon cancer .Survivin shRNA and APC double gene co-expression lentiviral vector was constructed to observe whether it could be successfully expressed in HT-29 colon cancer cells and whether it could impact on colon cancer cell apoptosis . Methods We selected the best shRNA interference fragment from the construc-tion of three pairs of complementary shRNA fragments and connected it with the effective fragment of APC ( aa1020-1698 ) to construct a double gene co-expression lentiviral vector .HT-29 cells were divid-ed into experimental group , empty loading group and blank group .HT-29 cells were observed by fluorescence microscopy after infec-tion.Survivin and APC expression levels were observed by real time PCR and western blot .Apoptosis was detected by caspase-3 activi-ty assay. Results ①We successfully constructed three pairs of shRNA sequences and proved that they had no human gene homolo -gous to the rest.Real time PCR analysis showed that the best sequence was shRNA 3.②After the sequence alignment of constructed shRNA vectors, three pairs of shRNA sequences were completely the same with the first designed sequence .Green fluorescence was observed in HT-29 cells by fluorescence microscope .The survivin content in experiment group (31.71 ±1.49) was significantly de-creased compared with empty loading group (100 ±0) and blank group(95.12 ±2.15)(P<0.05).The expression level of APC mR-NA was significantly increased compared with empty loading group (0 ±0) and blank group(0.51 ±0.15)(P<0.05).③The relative ratio of apoptosis in experiment group (0.573 ±0.050) was significantly increased compared with empty loading group (0.390 ± 0.040) and blank group(0.407 ±0.030)(P<0.05). Conclusion We have successfully constructed survivin-shRNA-APC double gene co-expression lentiviral vector which can be successfully expressed in HT-29 colon cancer cells , providing references for subse-quent gene therapy by the use of the carrier .
3.Effects of survivin shRNA-APC double gene co-expression lentiviral vector on subcutaneous xenotransplanted tumor of human HT-29 colon carcinoma in nude mice
Xixian YUAN ; Chao WEN ; Ya CAO ; Jingfeng DU ; Kai CHENG ; Yanli ZHU ; Houyu DUAN
Journal of Medical Postgraduates 2017;30(6):584-590
Objective Post-transcription RNA interference (RNAi) is more and more widely applied in multigene therapy.This study aimed to establish an subcutaneous xenotransplanted tumor (SXT) model of human HT-29 colon carcinoma in nude mice and investigate the effects of the survivin shRNA-APC double gene co-expression lentiviral vector on the growth of SXT.Methods Thirty-five nude mice were equally divided into five groups, double-gene survivin shRNA, survivin shRNA, APC, empty vector, and blank, and injected into the left anterior axillary with respective stably transfected cell lines and human HT-29 colon carcinoma cells, all at 2×106/mL, to establish an SXT model of human HT-29 colon carcinoma.The inhibition rate of tumor growth was calculated by measuring the size and weight of the SXT, the expressions of survivin mRNA and protein in the tumor tissue detected by real time PCR and immunohistochemistry respectively, and the apoptosis of the HT-29 colon carcinoma cells determined by TUNEL.Results The mean size and weight of the SXT were significantly reduced in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups as compared with the blank and empty vector groups (P<0.05), though increased in the survivin shRNA and APC groups in comparison with the double-gene group (P<0.05).The expressions of survivin mRNA and protein in the tumor tissue were remarkably lower in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups than in the blank and empty vector groups (P<0.05), even lower in the double-gene group than in the survivin shRNA, and APC groups (P<0.05).The apoptosis rate of the HT-29 colon carcinoma cells was markedly up-regulated in the double-gene survivin shRNA-APC ([56.72±3.17]%), survivin shRNA ([33.64±2.03]%), and APC groups ([31.19±1.79]%) as compared with the blank ([9.89±0.31]%) and empty vector groups ([10.06±0.43]%) (P<0.05), even more significantly in the double-gene than in the survivin shRNA and APC groups (P<0.05).Conclusion The survivin shRNA-APC double gene co-expression lentiviral vector can reduce the expression level the survivin gene, promote the apoptosis of colon carcinoma cells, and suppress the growth of the subcutaneous xenotransplanted tumor.
4.Use creatinine reduction ratio to predict the graft function recovery after kidney transplantation
Yun OUYANG ; Bingyi SHI ; Yeyong QIAN ; Hongwei BAI ; Jingyuan CHANG ; Yuan DU ; Yu FAN ; Jingfeng JIA ; Yawei WANG
Chinese Journal of Urology 2008;29(8):544-546
Objeetive To discuss the correlation of creatinine reduction ratio(CRR2)from posttransplant day 1 to day 2 and early graft function recovery status after kidney transplantation. Methods Clinical data of 80 patients after renal transplantation from Jan 2005 to Mar 2007 were retrospectively analyzed.Patients were divided into three groups according to the post-operative serum creatinine level:53 patients within IGF group[cereatinine<265.2 μmol/L by post-operative day(POD)no.5],14 patients within SGF group(creatinine>265.2 μmol/L on POD no.5,but no need for dialysis),and 13 patients within DGF group(need for dialysis in the first week post-transplant).Then the value and 99%CI of CRRz of these three groups were calculated. Results The value of CRR2 of IGF,SGF and DGF was(46.8±14.6)%,(25.6±13.5)%and(0.7±17.7)%respectively.And CRR2 99%CI of IGF,SGF and DGF was 41%-52%,15%-36%and-14%-16 0A respectively.There was significant difference in the value of CRR2 among IGF,SGF and DGF group.So a criteria for early diagnosis of IGF,SGF and DGF by CRR2 99%C1 was established:IGF(CRR2≥40%),SGF (15%<CRR2<40%)and DGF(CRR2≤15%). Conclusion CRR2 has a good correlation with early graft function recovery after kidney transplantation,and can be used to predict the occurrence of SGF and DGF.
5.Establishment and evaluation of artificial intelligence image marking method for magnetically controlled capsule gastroscopy
Lijuan FENG ; Lin TIAN ; Qian ZOU ; Zhongming DAI ; Xiaojuan TIAN ; Gongli YANG ; Jingfeng DU ; Mengqi XIANG ; Yu MENG ; Long XU
Chinese Journal of Digestion 2022;42(1):14-18
Objective:To explore the marking method for magnetically controlled capsule gastroscopy (MCCG) pictures with artificial intelligence (AI), so as to improve the work efficiency of endoscopist and to reduce the blind area of AI image reading.Methods:According to the consensus of MCCG, 24 parts of stomach in 14 775 pictures of MCCG from 35 subjects in Shenzhen Zifu Medical Technology Co., Ltd received MCCG from March to August, 2020 were marked by ten gastroenterologists and one developer of MCCG with medical background, the marking shape included rectangles and polygons. Among the ten gastroenterologists, three were senior endoscopist (the total number of gastroenteroscopy operations over 80 000, chief physician or associate chief physician), four were medium seniority endoscopist (the total number of gastroenteroscopy operations between 10 000 and 80 000, associate chief physician), and three were junior endoscopist (the total number of gastroenteroscopy operations less than 10 000, attending physician). The pictures of the same subject were pre-marked by two selected senior endoscopists with blind method, and the standard of marking with most appropriate coincidence rate was determined. The qualified marked pictures were automatically learn with AI deep learning method, and the learning results were fed back. Chi square test was used for statistical analysis.Results:According to the pre-marked results, the standard of coincidence rate for rectangular marking area was set as 50.0% and that for polygon marking area was 70.0%. The first correction for qualified rate was 39.0% (5 762/14 775). A total of 9 013 pictures were corrected. After repeated training and correction for one to five times, all pictures were qualified marked. The marking qualified rate of senior endoscopist partners was higher than that of partners of different qualifications (48.7%, 1 200/2 466 vs. 19.0%, 825/4 337), and the difference was statistically significant ( χ2=659.20, P<0.001). There was no statistically significant difference in the marking qualified rate between the senior endoscopist partners and partners of senior endoscopist and capsule developer (48.7%, 1 200/2 466 vs. 49.6%, 1 496/3 019; P>0.05). Conclusions:Establishment of AI marking method for MCCG can provide technical support for AI non-blind area reading, and AI non-blind area monitoring during the operation of MCCG.
6.Study on Quality Evaluation of Didang Qigui Decoction by HPLC Fingerprint Combined with Multi-component Content Determination
Yijia GUO ; Du CHENG ; Xiao ZHANG ; Liyan LEI ; Yanni LIANG ; Zheng WANG ; Jingfeng YANG
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(3):132-137
Objective To establish an HPLC fingerprint of Dingdang Qigui Decoction and analyze and evaluate it using chemical pattern recognition technology;To determine the contents of 5 effective chemical components in Dingdang Qigui Decoction;To provide a basis for its quality control.Methods The analysis was performed on Agilent 5 TC-C18(2)column(250 mm×4.6 mm).The mobile phase comprised of acetonitrile-0.1%phosphoric acid aqueous solution with the gradient elution at a flow rate of 1.0 mL/min.The detection wavelength was set at 260 nm.The column temperature was maintained at 30℃and the injection volume was 10 μL.SPSS 26.0 and SIMCA 14.1 were used to perform clustering analysis and principal component analysis on the 10 batches of Didang Qigui Decoction.The landmark components for inter batch differences were selected through orthogonal partial least squares discriminant analysis(OPLS-DA).Results The HPLC fingerprint with eighteen common peaks of Didang Qigui Decoction in 10 batches of sample was established,and the similarities of samples were between 0.828 and 0.989.Five indicative components were identified and quantitatively analyzed by comparing with the reference substances,which were paeoniflorin,mauroisoflavone glucoside,hesperidin,cinnamaldehyde and aloe rhodopsin.The linear ranges was 10.000 0-320.000 0 μg/mL,2.500 0-80.000 0 μg/mL,10.000 0-320.000 0 μg/mL,10.000 0-320.000 0 μg/mL,0.078 1-5.000 0 μg/mL,respectively,and their mean recovery ranged from 100.30%to 104.09%.Clustering analysis and principal component analysis divided 10 batches of samples from Didang Qigui Decoction into 2 categories.Through OPLS-DA screening,hairy pistil isoflavone glycosides,paeoniflorin,and hesperidin were selected as landmark components for quality differences.Conclusion The quality evaluation method for Didang Qigui Decoction established in this study is simple,sensitive,accurate,and reproducible,which can provide a basis for the quality evaluation of Didang Qigui Decoction.
7.Fusion expression of Asia I type FMDV neutralizing epitope with heavy chain constant region of sheep IgG and the assessment of its immunogenicity.
Jingfeng WANG ; Junjun SHAO ; Jing LI ; Shandian GAO ; Junzheng DU ; Guozheng CONG ; Tong LIN ; Huiyun CHANG
Chinese Journal of Biotechnology 2010;26(4):454-461
VP1 is a major antigenic protein of foot-and-mouth disease virus(FMDV), which induces the immune response against FMDV infection, and contains several epitopes of the virus. We designed and chemically synthesized a DNA fragment which encoding a tandem repeat protein of 136-160aa and 198-211aa of a strain of type Asia I FMDV, and cloned the gene of heavy chain constant region of sheep IgG. By using the BamH I, EcoR I and Xho I sites, both genes were cloned into pPROExHTb vector in turn to form a recombinant plasmid pPRO-FshIgG A chimeric protein, named FshIgG, was obtained after transforming the pPRO-FshIgG into Escherichia coli BL21 (DE3) host cell and induced by IPTG. Inoculation with 100 microg FsIgG induced strong neutralizing antibody response in guinea pigs, and FshIgG inoculated guinea pigs were also protected against 200 ID50 FMDV challenge. Our study indicated that the heavy chain constant region of sheep IgG can act as the carrier protein for FMDV peptide epitopes, and FshIgG is a potential multiepitope peptide vaccine candidate to prevent FMDV infection.
Animals
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Antibodies, Viral
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blood
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Capsid Proteins
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biosynthesis
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genetics
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Epitopes
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immunology
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Escherichia coli
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genetics
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metabolism
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Foot-and-Mouth Disease Virus
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genetics
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immunology
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Genetic Vectors
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genetics
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Guinea Pigs
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Immunization
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Immunoglobulin G
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genetics
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Immunoglobulin Heavy Chains
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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immunology
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Sheep
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Viral Vaccines
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genetics
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immunology