1.Research progress of peptide drugs for treatment of autoimmune diseases
Xuefeng WANG ; Liyang DONG ; Jun WANG ; Hongchang NI ; Chuan SU
Chinese Pharmacological Bulletin 2014;(5):732-734,735
Recent studies have found that peptide therapies tar-geting specific epitopes can avoid nonspecific immune suppres-sion induced by traditional medicines for the treatment of autoim-mune diseases, and have shown great therapeutic effect in ani-mal models of autoimmune diseases and clinical trials. The pa-per summaries the research progress and trends of peptide drugs for the treatment of autoimmune diseases from candidate peptide sources and their suppression mechanisms, which can provide a theoretical basis for the in-depth understanding of immune toler-ance and allow for discovery of new treatment for autoimmune diseases.
2.Study on the Transmission of Toxoplasma gondii by Semen in Rabbits
Shiguo LIU ; Chuan QIN ; Zhijun YAO ; Dong WANG
Chinese Journal of Parasitology and Parasitic Diseases 2006;24(3):166-170
Objective To confirm the transmission of Toxoplasma gondii by semen and to investigate the impact of vaginal status on the transmission of T. gondii in female rabbits. Methods Sixteen male rabbits were infected with T. gondii by intraperitoneal injection each with 1 ×105 RH tachyzoites. Eight rabbits died in 8-14 d after infection.Artificial vagina was used to collect semen from male rabbits weekly before and after infection for 8 weeks. If more than 2 portions of semen from 8 survived male rabbits were collected after infection, the collected semen was mixed weekly for later use. Twenty-seven female rabbits were divided into 4 groups: group 1 with normal vagina (7 rabbits), group 2with wounded vagina (7), group 3 with trichomonas vaginitis (7) and group 4 with colpomycosis infection (6). Tachyzoites were found in mixed semen digested by trypsinase, and were used for endovaginal artificial insemination to female rabbits by uterine cavity tube once a week for 8 consecutive weeks. 2-3 d after every insemination, 2 ml blood was collected from helix vein of each rabbit, and stored at -40 ℃ for use. Anti-T. gondii antibody was examined by ELISA and the B1 gene of T. gondii was detected by PCR. Results Anti-T. gondii antibody was detected in some rabbits (2, 3, 1, and 1 rabbits from each of the groups respectively) on the 16th day after the first insemination. The positive rate of ELISA was 25.9%. The amplification of B1 gene (200 bp) by PCR appeared positive from the blood samples on the 3rd day after the first insemination and the last positive one was proved on the 51th day after the first insemination.Number of positive samples was 2, 1, 3 and 1 in the 4 groups respectively, with an overall PCR positive rate of 18.5%.Only 3 of the 27 rabbits were positive by both ELISA and PCR. Conclusions T. gondii can be transmitted by semen and the health status of vagina shows no impact on it.
3.Significance of ST/TP Relative Value in Electrocardiogram in Diagnosis of Hypocalcemia
cai-xia, CHEN ; shou-chuan, WANG ; na, DONG ; ye, SHAN
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To evaluate the significance of ST/TP relative value in electrocardiogram in diagnosis of hypocalcemia.Methods ST/TP relative value in electrocardiogram in 61 children with hypocalcemia and 31 normal children were detected by NIHOH KOHDEN CardiofaxQ ECG-9130P electrocardiogram machine designed by Japanese futian company.Calcium ion was checked out by clinic electrolyte equipment.Results There was significant difference of ST/TP relative value between patients with hypocalcemia and normal children(P
4.Comparison of rabbit retinal fixation techniques: chemical fixation and microwave irradiation
Chao, QU ; Dan, WANG ; Wen-dong, WANG ; Ying-chun, LI ; Dan-dan, DONG ; Ying-chuan, FAN
Chinese Journal of Experimental Ophthalmology 2011;29(1):17-20
Background Conventional chemical method to fix the entire eyeball results in the edema of retinal nerve fiber layer and therefore influent the evaluation of tissue structure.Research showed that microwave irradiation can avoid this phenomenon. Objective This study was to investigate the appropriate energy and time duration of microwave irradiation for fixation of rabbit eyes specimen and compare the influence of different fixation methods on retinal structure. Methods Twenty-two New Zealand white rabbits were sacrificed by using venous air embolism.The eyeballs were enucleated from the rabbits and immersed in chemical reagents with glacial acetic acid,methyl aldehyde and chloroform for 2 days as the control group.The eyeballs were immersed in 400 ml physiological balance solution immediately,and then were fixed by microwave irradiation for 80,160,240 and 320 seconds under the lower power as the microwave irradiation groups.In the microwave irradiation+chemical fixation group,the specimens were immersed in chemical reagents for 1-2 hours after the microwave radiation.The morphology of retinal structure was examined and area of retinal detachment was calculated by hemotoxylin and eosin staining under the light microscope. Results A better fixation effect was obtained in the retinal section of microwave radiation for 240 seconds in comparison with that of the traditional chemical reagent fixation method for 2 days.The retina detachment ratio was 16.3%±11.5% for simple microwave method,50.0%±24.5% for chemical method,and 6.7%±7.8% for microwave+chemical method.showing a significant difference among 3 groups(F=32.43,P=0.000).Retinal staining was clear and retinal structure was almost normal in the specimens of microwave+chemical fixation group with 200-240 seeonds microwave irradiation and 1-2 hours chemical fixation. Conclusion Microwave irradiation method is a more ideal way for the fixation of retina because of taking short duration,lower toxicity and better staining.The combination of microwave radiation and chemical reagents can acquire an excellent quality of retinal section.
5.Clinic study of plasma radiofrequency at low temperature in tonsillectomy.
Jie WANG ; Chuan DONG ; Chuan-yu LIANG ; Qihong FU ; Zhenhua JIANG ; Liling CHEN ; Li XIA
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005;40(5):382-383
Adolescent
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Adult
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Child
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Child, Preschool
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Electrocoagulation
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methods
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Female
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Humans
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Hypothermia, Induced
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Male
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Middle Aged
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Prospective Studies
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Tonsillectomy
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methods
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Young Adult
6.Expression of mieroRNA-224 in HepG2 cells and analysis of its predicted target genes
Qiong LI ; Ge WANG ; Jinlu SHAN ; Chuan CHEN ; Zhimin ZHANG ; Wen XU ; Xizhong LUO ; Dong WANG
Chinese Journal of Digestive Surgery 2008;7(4):297-299
Objective To investigate the expression of microRNA-224 in HepG2 cells and analyze its target genes to reveal its role in the carcinogenesis of hepatoma. Methods The genes with differential expression in HepG2 cells and LO2 cells were obtained by gene expression microarray analysis. The up-regulated target genes of microRNA-224 were predicted by bioinformatics method, and their functions were analyzed. Results Compared with LO2 cells, microRNA-224 was highly expressed in HepG2 cells. A total of 264 target genes of microRNA-224 were predicted, including genes involved in cell cycle, signal transduction, cell differentiation, proliferation and apoptosis. Conclusions MicroRNA-224 is highly expressed in HepG2 cells. MicroRNA-224 plays an important role in the carcinogenesis of hepatoma via regulating the expression of its target genes directly or indirectly.
7.Effects of astragaloside Ⅳ on electrocardiogram and action potential of ventricular myocytes in guinea pigs
Yun, ZHOU ; Jun-jie, WANG ; Wei, ZHAO ; Chuan, ZHANG ; Wei-dong, ZHANG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(6):677-680
Objective To observe the effects of astragaloside Ⅳ on electrocardiogram (ECG) and action potential of ventricular myocytes in guinea pigs. Methods ECG was recorded in vivo and ex vivo by using conventional ECG recording method from anesthetic guinea pigs and Langendoff perfusion model of hearts. Action potentials were recorded from isolated papillary muscles of right ventricles of guinea pigs by using microelectrode techniques. Results RR interval was prolonged by Astragaloside Ⅳ in a dose-dependent manner both in vivo and ex vivo. Astragaloside Ⅳ shortened action potential duration (APD), while had no effects on resting potential, action potential amplitude and maximal rate of depolarization. Conclusion Astragaloside Ⅳ exerts a negative chronotropic effect on heart and shortens APD of cardiac myocytes, which may be involved with calcium channels.
8.Expression, purification and identification of Abeta fusion protein with enhanced solubility.
Rui ZHU ; Chuan-dong ZHENG ; Fang WANG
Journal of Southern Medical University 2010;30(3):447-450
OBJECTIVETo economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.
METHODSThe cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.
RESULTSThe result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.
CONCLUSIONMBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.
Alzheimer Disease ; genetics ; Amyloid beta-Peptides ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Maltose-Binding Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification
9.Isolation and purification of anaphylactic antibody from guinea pig and preliminary research on its role in passive cutaneous anaphylaxis test (PCA)
Na DONG ; Lei CHEN ; Miao XU ; Haiqing DENG ; Cheng SU ; Chuan QIN ; Guozhi WANG
Chinese Journal of Microbiology and Immunology 2010;30(2):169-174
Objective To establish guinea pig model of type Ⅰ anaphylaxis, isolate the anaphy-lactic antibody(IgE and IgG) preliminarily from sera of sensitized guinea pigs, and investigate its functional characteristics. Methods Animal model was established by sensitizing guinea pigs with OVA and Al(OH)_3, level of antibody was determined by ELISA, IgE and IgG in sera were preliminarily isolated through saturated ammonium sulphate precipitate and affinity chromatograph of Protein A. A continuous passive cutaneous ana-phylaxis test (PCA) was performed by sensitizing guinea pigs individually with IgE and IgG and then challenging at different time , and the variation of blue spots in skin were observed after challenge . Results Concentration of IgE in model group and control group were 719.3750 ng/ml and 2.5250 ng/ml, the optical density of IgG in model group and control group were 0.9921 and 0.0174, the level of two antibodies in model group were significantly higher than that of control group (P<0.05). In 9 d continuous PCA test, the blue spot induced by IgE in skin lasted for 9 days and appeared the largest when challenged at day 5. The diameter of blue spot induced by IgG was the largest when challenged at day 2 and then decreased fast. Con-clusion Anaphylactic antibodies were successfully preliminarily isolated from sera of sensitizing guinea pig, both IgE and IgG play roles in type Ⅰ anaphylaxis of guinea pig, the hypersensitive reaction induced by IgG is fast and short than that induced by IgE, and IgG may become an important surrogate marker in immunotoxic-ity evaluation(type Ⅰ anaphylaxis)of vaccine.