The Expression of Galectin-7 in Bronchial Mucosa of Asthmatic Children and Its Effect on the Apoptosis of Bronchial Epithelial Cells / 现代生物医学进展

Objective:To investigate the expression of Galectin-7 in the bronchial mucosa of asthmatic children and its effect on the apoptosis of bronchial epithelial cells.Methods:Bronchial mucosa of asthmatic children and children with bronchial dilation were collected and the expression of Galectin-7 was detected by Western blot.Human bronchial epithelial cells were cultured in vitro,the cells were transfected with Galectin-7 siRNA to interfere the Galectin-7 expression,while siRNA control was transfected as the control group.The experiment was divided into normal group,control group,infected group and experimental group.The normal group was normal human bronchial epithelial cells,the cells in the control group was transfected with siRNA control,the infected group was infected with RSV,the experimental group was transfected with Galectin-7 siRNA and infected with RSV.After 24h culture,Galectin-7 protein expression and cell apoptosis were detected in the cells of each group.Western blot was used to detected the expression of Bcl-2,Bax,STAT3 and p-STAT3.Results:The expression of Galectin-7 in bronchial mucosa of asthmatic children was significantly higher than that of the non asthmatic children (P＜0.01).There was no significant difference in the Galectin-7 level between the normal group and the control group (P＞0.05).The levels of Galectin-7,Bax and apoptosis in the infected group were significantly higher than those in the normal group,while the levels ofp-STAT3 and Bcl-2 were significantly lower than those in the normal group (P＜0.01).The levels of Galectin-7,Bax and apoptosis in the experimental group were significantly lower than those in the infected group,while the levels of p-STAT3 and Bcl-2 were significantly higher than those in the infected group (P＜0.01).Conelusions:The expression of Galectin-7 was up-regulated in the bronchial mucosa of asthmatic children,which might promote the apoptosis of bronchial epithelial cells by activating STAT3.

Localization and identification of the mutant gene in the loop-tail mouse / 中国比较医学杂志

Objective To define the loci of the mutant gene in the loop-tail mouse.Methods To study the heredity pattern, loop-tail mice were mated with normal C57BL/6J and C3H mice.Their offsprings with loop-tail or normal phenotype were registered respectively.Microsatellite marker D1Mit113 and D1Mit149 were used to locate the mutant gene.Based on fine mapping, the candidate gene Vangl2 was found.Vangl2 gene from the loop-tail mice was amplified by PCR followed by sequencing.Incision enzyme FspBI ( BfaI ) identified the genotype of offspring from loop-tail mice intercrossing.Results Heredity test indicated that the loop-tail phenotype was controlled by a single dominant gene not with 100%penetrance but was affected by genetic background.A C-to-T transversion was at the 1345bp in Vangl2 gene of the loop-tail mice.Conclusions The C-to-T transversion introduces a pre-termination codon of amino acids and causes the phenotype of loop-tail phenotype.None homozygous mice were found in the offsprings, suggesting that the homozygous mice are lethal.

Production of a corneal opacity mouse by N-ethyl-N-nitrosourea mutagenesis and chromosome mapping of the mutant gene / 中华实验眼科杂志

Background N-ethyl-N-nitrosourea (ENU)-induced mouse mutagenesis is a powerful approach for the study of gene function and the generation of human disease models.Objective This study was to create the corneal morphologic change and map the mutant gene of a kind of corneal opacity in ENU mutagenesis in mouse.Methods ENU was intraperitoneally injected in forty C57BL/ 6J (B6) male mice aged 8-10 weeks old.The male mice were mated with the same strain female mice.Their progenies were screened for visible eye mutation,and the mutant mice were mated with the same strain mice to confirm the heredity of mutation phenotypes.Hematoxylin & eosin staining was used to examine the histopathological change of cornea in one mouse with ENU-induced corneal opacity.To map the mutant gene,[(B6×D2)F1 ×B6] N2 mutant mice were bred,and the genome of the N2 mice was scanned by microsatellite markers distributed equally on the mouse chromosome.The microsatellite linked to the mutant gene was determined by the log odds score.This experimental procedure was approved by Ethic Committee about Experimental Animal Care and Use of Yangzhou University.Results The founder mouse,which was the progeny of an ENU-treated B6 male mouse and an untreated B6 female mouse,had a corneal opacity phenotype.After mating the mutant with B6 mice,19 of 59 descendants appeared corneal opacity phenotype.Thickening of corneal stroma,neoangiogenesis,infiltration of inflammatory cells and proliferation of fibroblasts were exhibited in cloudy cornea in ENU-induced mutated mice under the optical microscope.After linkage analysis between microsatellite markers and the mutant gene,the mutant gene was linked to D2Mi307,which was located at 63.42 cM.Three cases of 26 N2 mice underwent recombination with the LOD 3.79.The mutant gene associated with the cornea phenotype was located on chromosome 2.Conclusions This study map the mutant gene associated with the cornea phenotype on chromosome 2.The strain might be used as a mouse model for heritable human corneal opacity.

Prolonged hypotensive effect of human tissue kallikrein gene delivery and recombinant enzyme administration in spontaneous hypertension rats

To evaluate the feasibility of treating hypertension by human tissue kallikrein gene (KLK1) delivery and by enzyme (rK1) administration, two recombinant vectors expressing KLK1 cDNA were constructed for gene delivery (pcDNA-KLK1) and recombinant enzyme preparation (pOV-KLK1). Expression of the pcDNA-KLK1 vector in COS-1 cells was confirmed by immunofluorescence and in spontaneous hypertension rats (SHR) by enzymatic detection. Following intramuscular or intravenous injection with the pcDNA-KLK1 vector, systolic pressure of SHR was significantly decreased, which lasted for 20 d to two months depending on dose, route and/or time of injection. Egg white containing recombinant hK1 was prepared by injection of egg-laying hens with the oviduct-specific expression vector pOV-KLK1 and administered into SHR via oral gavage. Following administration, systolic pressure of the SHR was decreased to that of normal rats, which lasted for 3-5 d depending on the dosage used. These data suggest that both hKLK1 gene delivery and recombinant enzyme administration can be used as alternative strategies for treating human hypertension.