1.Glucagon-Like PeptideⅠand Targeted Differentiation of Stem Cells
Lei YAN ; Xiao-Ling MU ;
China Biotechnology 2006;0(04):-
Diabetes mellitus has become one of the diseases which threaten the heath of human being in the 21st century.A goal of research in diabetes is to find a way to increase the number of functional insulin-producing cells. Islet transplantation has been considered to be the most effective approach to cure type Ⅰ and part of type Ⅱ diabetes mellitus.This approach, however, is severely limited by an inadequate supply of donor islets available for transplantation.Moreover, recent progress of stem cells research has shown that stem cells may act as a new source of islet transplantation in diabetes mellitus treatment. Recent evidence indicates that Glucagon-Like PeptideⅠ(GLP-1) plays a very important role in targeted differentiation of stem cells into Insulin-Producing Cells and pancreatic development. GLP-1 is an intestine-derived insulinotropic hormone that stimulates glucose dependent insulin production and secretion. GLP-1 can induce differentiation of stem cells into insulin-producing cells, which is achieved by up regulation of PDX-1 expression.PDX-1 is a transcription factor critical for pancreatic development and endocrine cell neogenesis and a marker for pancreatic stem cells. These new findings suggest an approach to create Insulin-Producing cells in vitro by expanding stem/progenitor cells and then to convert them into Insulin-Producing cells by treatment with GLP-1. Thus GLP-1 may be a means by which to create Insulin-Producing cells ex vivo for transplantation into patients with insulinopenic type Ⅰ diabetes and severe forms of type Ⅱ diabetes. This article reviews recent progress about GLP-1 and targeted differentiation of stem cells induced by GLP-1.
2.Mesenchymal chondrosarcoma of the orbit:a case report
Lei SUN ; Ling QIU ; Lei YU ; Yan MA ; Xiaojing JIA
Practical Oncology Journal 2014;(4):342-343
Mesenchymal chondrosarcoma originated in the primitive mesenchymal tissue .It usually devel-ops in the short bones such as hand ,foot and body bone ,while extremeIy rare in the orbit .We report a case of mesenchymal chondrosarcoma of the orbit which is confirmed by pathology .
3.Transfection of embryonic stem cells with green fluorescent protein gene and their differentiation into neural cells
Zhi-yan, SHAN ; Jing-ling, SHEN ; Lei, LEI ; Yan-ning, XU ; Lian-hong, JIN
Chinese Journal of Endemiology 2008;27(4):397-400
Objective To establish embryonic stem cells (ESC) that can express green fluorescent protein (CFP) and differentiate them into neurons. It would provide tagging neurons for clinical transplantation to cure neural system diseases. Methods ESC (R1) was transfeeted with a plasmid containing the GFP by electroporation. A transgeuic cell line was obtained after selection with G418. The ESCs were characterized by AKP staining. Monolayer differentiation method was used to induce neural differentiation derived from GFP-ESC and immunofluorescence method was used to identify Tuj1 positive cells. Results There was no significant difference(X2=3.14,P0.05) in transfect rates between liposome and electroporation (65% vs 79%). The AKP staining of GFP-ESC was positive. GFP-ESC could be differentiated into neural cells. Conclusions These results show that ESC expressing GFP has been estabhshed, which can be differemiated into neurons.
6.Therapeutic effect of phenylethanoid glycosides on cyclopfosphamide-induced dyszoospermia in mice and its mechannism
Donghai ZHAO ; Lei ZHANG ; Yan ZHANG ; Ling QI ; Xiangming ZOU
Journal of Jilin University(Medicine Edition) 2014;(3):612-615
Objective To explore the therapeutic effect of phenylethanoid glycosides on cyclophosphamide (CTX)-induced dyszoospermia in mice and to preliminary elucidate the mechanisms involved in the process. Methods Phenylethanoid glycoside was extracted by ethanol extraction.Forty male BALB/C mice were randomly divided into control group,model group,low dose of phenylethanoid glycosides group (50 mg· kg-1 )and high dose of phenylethanoid glycosides group (100 mg·kg-1 ).Except control group,the dyszoospermia mouse model was established by peritoneal injection of CTX at the daily dose of 80 mg· kg-1 ,once daily for successive 5 d. After modeling, phenylethanoid glycosides were intragastrically administered at corresponding doses to each phenylethanoid glycosides group.Equal volume of normal saline was given to the mice in control group and model group by gastrogavage.All the medication was performed once daily for successive 30 d.The testis tissue was obtained 24 h after the last intragastric administration.The level of testosterone in the testis tissue homogenate was determined by enzyme linked immunosorbent assay.The sperm counts, the motility rates, and the teratospermia rates in various groups were compared.The morphological changes of the testis tissue were observed using HE staining.Results Compared with control group, the sperm count and the motility rate were decreased, the teratospermia rate was increased,and the testosterone level in the testis tissue homogenate was decreased in model group(P<0.01).Compared with model group,the sperm counts and the motility rates were increased,the teratospermia rates were decreased, and the testosterore levels in the testis tissue homogenate were increased in phenylethanoid glycosides groups (P<0.05 or P<0.01).The histological results showed atrophy and degeneration of seminiferous tuble,thicker seminiferous epithelium and azoospermic lumina in model group;the number of seminiferous epithelial layers was increased and the seminiferous cells orderly arranged, and many sperms were found in the tubules in phenylethanoid glycosides groups.Conclusion Phenylethanoid glycosides has obviously therapeutical effect on CTX-induced dyszoospermia in mice,and its mechanisms might be correlated with recovering the testosterone level.
7.Effects of Probucol on Expression of Bcl-2,Bax and Oxidative Stress in Myocardial Infarcted Rats
Shu-Xian ZHOU ; Yu-Ling ZHANG ; Yan ZHOU ; Juan LEI ;
Chinese Journal of Hypertension 2006;0(10):-
Background Cardiomyocytes apoptosis was related to oxidative stress which has been shown to play an important role in ventricular remodeling after myocardial infarction(MI).Probucol is a hypolipidemic and antioxidant agents.Several reports demonstrated its cardioprotective effect after MI.However,the exact effect of probucol on the modification of the apoptosis related gene Bcl-2 and Bax is not clear.Objective To investigate the effects of probucol on mRNA and protein expression of Bcl-2,Bax and oxidative stress in myocardial infarcted rats.Methods Forty-one SD rats that survived 24 h after ligating left anterior descending coronary artery were randomly to receive placebo-saline(5 mL/d,n=20)or probucol(probucol 60 mg/kg?d,n=21).Twelve rats underwent sham operation were served as control(n=12).Six weeks after treatment,hemodynamic pararmeters and left ventricular function were measured with catheterization.Cardiomyocytes apoptosis were determined by TUNEL method.Myocardium mRNA of Bcl-2 and Bax expression levels in the non-infarcted myocardium were as- sessed by RT-PCR.The myocardium protein expression of Bcl-2 and Bax in the non-infarcted myocardium were de- termined by Western blot.Colorimetry was used to determine oxidative metabolism index in myocardium.Results 1)Compared with the sham rats,all MI rats showed marked decrease in Bcl-2 mRNA and protein expression in myo- cardium with increase of Bax mRNA and protein expression and apoptosis index(P
8.Expression and promoter methylation of Kras gene in thymic lymphomas induced by ionizing radiation
Lei YU ; Ling QIU ; Lei SUN ; Yan MA ; Guizhi JU ; Xiaojing JIA ; Yugang GAO
Journal of Jilin University(Medicine Edition) 2014;(6):1127-1131
Objective To study the changes of mRNA and protein expressions of Kras gene in thymic lymphomas induced by ionizing radiation,and to detect the methylation of CpG islands in promoter region of Kras gene,then to investigate the mechanisms for the occurrence of radiation carcinogenesis.Methods The thymic lymphoma models of BALB/c mice were made by X-ray irradiation,then the total RNA was extracted,cDNA was synthesized and the total protein was extracted from both thymic lymphoma tissue and normal thymus tissue;the mRNA and protein expressions of Kras gene in thymic lymphoma tissue and normal thymus tissue were detected by RT-PCR and Western blotting method, and the methylation of CpG islands in promoter region of Kras gene was detected by bisulfite sequencing PCR. Results The mRNA expression level of Kras gene in thymic lymphoma tissue was significantly higher than that in normal thymus tissue(P<0.01).The protein expression level in thymic lymphoma tissue was about 1.41 times higher than that in normal thymus tissue;4 CpG sites were methylated detected by bisulfite sequencing PCR in normal thymus tissue, however, 1 CpG site was methylated in thymic lymphoma tissue,the CpG islands in promoter region of Kras gene were demethylation state in thymic lymphoma. Conclusion Ionizing radiation can cause the changes of mRNA and protein expression levels of Kras gene in thymic lymphoma tissue by demethylation state of Kras gene,eventually lead to the occurrence of tumor;it might be one of the mechanisms for the occurrence of radiation carcinogenesis.
9.Rapid screening of 28 alkaloids in food poisoning samples by liquid chromatography-tandem mass spectrometry
ZHAO Ling-guo ; LUO Lan ; YIN Zhen-yi ; REN Yan ; LEI Lei ; MA Zhi-feng
China Tropical Medicine 2023;23(3):260-
Abstract: Objective To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.