1.Zebrafish as a model animal for the study of blood-brain barrier permeability by biomolecules.
Ailing FU ; Hengyu CHEN ; Xingran XU ; Baoquan ZHAO
Acta Pharmaceutica Sinica 2012;47(11):1447-51
Blood-brain barrier (BBB) is the major obstacle for drug delivery into the central nervous system (CNS). However, there is no ideal model animal for the study of BBB permeability till now. Currently zebrafish (Danio rerio) has emerged as a powerful model organism for the study of vertebrate biology. In this study, the feasibility of using zebrafish as model animal was investigated for BBB permeability by comparing the results of administration of BBB-penetrating peptide and protein to mouse and zebrafish. The results showed that the BBBs of mouse and zebrafish were similar in molecular permeability. Additionally, zebrafish has advantageous features as a model animal, such as small size, fertile and easy to breed. Therefore, it is suggested that zebrafish may be a favored model for the study of BBB permeability.
2.Cloning and Sequence Analysis of a Novel Stage-Specific cDNA from Adult Trichinella spiralis
Baoquan FU ; Feng WANG ; Xiuping WU ; Tingxian NIU ; Qiang LU ; Mingyuan LIU ; Boireau PASCAL
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To clone a stage-specific novel cDNA from 5 day-old adult worm (ADS) of Trichinella spiralis. Methods The cDNA library of AD5 was screened by an AD5 stage-specific cDNA probe labeled with digoxigenin (DIG). The positive clones were sequenced and analysed. Results The positive clone contained a cDNA insert of 1 132 bp in length with a full length open reading frame (ORF) of 1 032 bp. The cDNA encoded a polypeptide of 343 amino acid residues(aa) with a molecular weight of 35.1 kDa and an isoelectric point (IP) of 4.8. InterProScan analysis showed that the 117 - 120 aa (SGYG) was a glycosaminoglycan attachment site, 27- 86 aa was nematode cuticle collagen N-terminal domain and 153-228 aa was collagen repeat (G-x-y) domain. Signal PV2.0 analysis indicated that the region of 1-43 aa was a singal peptide. Blastn homology analysis in Genbank revealed that the cDNA had no obvious homology to any other known gene sequence. Blastp analysis revealed high homology to cuticle collagen with identities more than 40 % . Conclusion A novel ADS stage-specific cDNA encoding a full length ORF was cloned and sequence analysis showed this gene encoded cuticle collagen of Trichinella spiralis.
3.Detection of Anti-Trichinella spiralis Antibody by Indirect ELISA Using Recombinant Protein as Antigen
Tingxian NIU ; Feifei PAN ; Mingyuan LIU ; Qiang LU ; Baoquan FU ; Boireau PASCAL
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(03):-
Objective To study the specificity, sensitivity of diagnostic antigen for Trichinella spiralis(T. s). Methods T668 recombinant protein from highly efficient expression of newborn larvae stage-specific gene of T. s in E. coli as diagnostic antigen. By using negative and positive sera from rabbit, pig, human as the first antibody, and goat-anti-rabbit IgG, goat-anti-pig IgG, goat-anti-human IgG labeled with HRP as the secondary antibody, indirect ELISA method was established for detecting anti-T.s antibody, while excretory-secretory (ES) antigen of T.s muscle larvae was used as control. Results Sera from rabbit, pig and human were detected by T668 recombinant protein as antigen, which showed a positive rate of 100% with 0.016 ?g/well. There was no difference on the results between the T668 recombinant antigen and the ES antigen for diagnosing T. s-infection. Conclusion The T668 recombinant antigen is promising in substituting ES antigen in the detection of anti-T. s antibody.
4.Generation of cytochrome P-450 CYP3A65 labeled fluorescence transgenic zebrafish and its biological response to environmental pollutants
Chunjie LL ; Jian ZHAO ; Shiyong ZHANG ; Weitong PAN ; Yunzhu PU ; Qiyan JLA ; Xiaodan ZHA ; Yannan SHANG ; Chunqian HUANG ; Yanqin LLU ; Yuxu ZHONG ; Qian LL ; Rigao DLNG ; Ailing FU ; Baoquan ZHAO
Chinese Journal of Pharmacology and Toxicology 2014;(6):870-877
OBJECTlVE To establish Tg(-6.3CYP3A65∶EGFP) transgenic zebrafish for quick, intuitive detection of heavy metals ( copper, cadmium and zinc) , dioxin-like PCBs ( PCB126) and other environmental pollutants. METHODS Tol2 transposon system was used to generate transgenic zebrafish lines Tg(-6.3CYP3A65∶EGFP) in which CYP3A65 promoter regualated labeled fluorescence. The effect of heavy mentals ( copper, cadmium and zinc ) and PCB126 on the relative amounts of CYP3A65 gene expression was determined by observing the change in fluorescence intensity. RESULTS The relative gene expression of CYP3A65 was significantly increased after 96 h exposure to copper 0.1 and 0.2μmol·L-1 , cadmium 0.35 and 0.7μmol·L-1 , zinc 1.5 and 3μmol·L-1 , and PCB126 2-32μmol·L-1 , respectively ( P<0.01) , but decreased after 96 h exposure to copper 0. 9 μmol·L-1 , cadmium 2. 7 and 5.4 μmol·L-1 , and zinc 24μmol·L-1 , respectively( P<0.01) . CYP3A65 gene expression was significantly increased after 168 h exposure to copper 0.1 and 0.2 μmol·L-1 , cadmium 0.35 and 0.7 μmol·L-1 , zinc 1.5 and 3 μmol·L-1, and PCB126 2-32 μmol·L-1, respectively(P<0.01), but decreased after 168 h exposure to copper 0.9 μmol·L-1, cadmium 2.7 and 5.4 μmol·L-1, and zinc 12 and 24 μmol·L-1( P<0.05) , in a concentration-dependent manner. CONCLUSlON The results suggest that zebrafish CYP3A65 gene expression and the CYP3A65 labeled fluorescence lines can be another candidate biomarker for detecting environmental pollutants.
5.Prediction of postoperative chemotherapy tolerance in patients with endometrial cancer by nomogram model
Ting FU ; Shunyu HOU ; Li SUN ; Ting LENG ; Baoquan LIANG ; Chen CHENG ; Gongzhao QIN
Chinese Journal of Postgraduates of Medicine 2020;43(12):1124-1129
Objective:To construct a nomogram model to predict the tolerance of postoperative chemotherapy in endometrial cancer patients by analyzing nutrition-related indexes.Methods:Patients with endometrial cancer who underwent surgery and postoperative chemotherapy in Suzhou Municipal Hospital from July 2013 to July 2019 were included in this study. Some nutrition-related indexes were collected, including age, diabetes, body mass index(BMI), blood albumin, prealbumin, lymphocyte count, hemoglobin, blood platelets, C-reactive protein and blood cholesterol, aspertate aminotransferase, alanine aminotransferase and chemotherapy tolerance. The model for prediction of postoperative chemotherapy tolerance of patients with endometrial cancer were constructed by analyzing these nutrition-related indexes.Results:A total of 83 patients were enrolled, aged from 32 to 81 years, with an average age of (55.15 ± 9.38) years, and BMI were 14.53 - 30.11 kg/m 2, with an average BMI of (22.03 ± 2.93) kg/m 2. All the 83 patients were confirmed as endometrial cancer by postoperative pathology, and received TP or TC chemotherapy after surgery. Before starting chemotherapy, 83 patients were scored on the NRS-2002 scale, 43 were considered to be at risk of malnutrition or nutrition. After receiving chemotherapy, 34 patients developed intolerance to chemotherapy, and the remaining 49 were tolerable. The results of univariate and multivariate regression analysis showed that the presence or absence of diabetes ( HR = 5.634, P = 0.038), lymphocyte count ( HR = 0.146, P = 0.004), and albumin ( HR = 0.857, P = 0.011) were independent influencing factors of chemotherapy tolerance. Conclusions:Diabetes mellitus, lymphocyte count and albumin level can be used to predict the tolerance of chemotherapy in patients with endometrial cancer. The nomogram model of these three nutrition-related indexeshad a good prediction ability to predict the tolerance of chemotherapy in patients with endometrial cancer.
6. Advances in research on echinococcus shiquicus tapeworm
Guoqiang ZHU ; Li LI ; Hongbin YAN ; Yantao WU ; Wenhui LI ; Baoquan FU ; Wanzhong JIA
Chinese Journal of Preventive Medicine 2019;53(1):112-117
Echinococcosis is an age-old disease that causes serious damage to the animal husbandry and the human health perennially. As a newly discovered species of