1.Incidence Rate of Overexpressing AmpC ?Lactamases in Cefoxitin-resistant Gram-negative Bacillus and Study on Drug-resistance and Genotype of Plasmid-mediated AmpC ?-Lactamases
Fuying FENG ; Wangping HU ; Xiangyue YANG ; Yabin ZHANG ; Xinlan HU ; Rongying GUO
Chinese Journal of Nosocomiology 2006;0(06):-
87%. CONCLUSIONS The surveillance of overexpressing AmpC ?-lactamases in cefoxitin-resistant Gram-negative bacillus must be enhanced.The therapy of infections caused by related bacillus should make imipenem and meropenem a chief choice.DHA-1,CMY-2 and CMY-22 AmpC enzymes are found in Fuzhou.
2.Antibody Preparation and Expression Analysis of a New Protein Mimecan in Pituitary Tumors
Sanmei HU ; Fengling CHEN ; Tianqin CHEN ; Tingjun YE ; Rongying LI ; Wenjing SHI ; Huaidong SONG ; Zhenyu LU
Chinese Journal of Biochemistry and Molecular Biology 2005;21(1):13-18
Mimecan belongs to a family of leucine-rich proteoglycans that are secreted into the extracellular matrix. In order to investigate the function of mimecan, the coding region of mimecan was amplifed from a human pituitary cDNA by PCR and the recombinant prokaryotic expression vector pGEX-M was constructed. The vactor was transformed into E.coli BL21(DE3)and the GST-M fusion protein of 38 Kd was ecpressed in the bacteria under induction of IPTG. After purification, the fusion protein was infucted into New Zealand rabbits to prepare polyclonal antibody. The antibody was tested by Western blotting for their specificity and sensitivity. Using the antibody it was found the mimecan was expressed highly in certain types of human pituitary tumor tissues. These results make it possible for studying the biological function of mimecan.
3.Vinblastine enhances the expression of tumor resistance gene abcb4 in zebrafish
Rongying HU ; Congjie SUN ; Lu JIN ; Sijia HE ; Yanhua ZHOU ; Liping SHU ; Zhixu HE
Basic & Clinical Medicine 2017;37(6):758-762
Objective To explore the effects of vinblastine on abcb4 tumor resistance gene expression in early development zebrafish and lay an important foundation for multi-drug resistant antineoplastic screening in zebrafish model.Methods Zebrafish embryos of 0.5~1.5 hours post-fertilization were exposured to different concentrations of vinblastine, and then calculated the IC50 of vinblastine.Zebrafish embryos of 0.5~1.5 hours post-fertilization were treated with different concentrations of vinblastine and embryo culture medium respectively, and then observed zebrafish embryo development in 24~120 hours post-fertilization and recorded the number of death, hatch and malformation.Evaluating the impact of vinblastine on abcb4 gene expression in zebrafish with quantitative real-time PCR and whole-mount in situ hybridization by collecting zebrafish embryos exposed to different concentrationsof vinblastine.Results Vinblastine IC50 in zebrafish embryos was 3.08 μmol/L.The mRNA level of abcb4 gene in vinblastine treated embryos was significantly increased compared to blank control group.Moreover, abcb4 gene positive hybridization signals were found in the small intestine of zebrafish embryos after 120 hours post-fertilization and also found in the brain and heart of zebrafish embryos by the method of whole-mount in situ hybridization.Conclusions Vinblastine can significantly increase the expression level of abcb4 tumor resistancegene in early development zebrafish embryos, which indicates that zebrafish can be used as a tumor resistant drug screening model.
4.Resistin gene overexpression in adipose tissue of SD rats with high-fat diet and its correlation with leptin and insulin
Shaoyun WANG ; Huaidong SONG ; Rongying LI ; Jinfeng TANG ; Fengying LI ; Libin ZHOU ; Ying YANG ; Renming HU ; Bo MOU ; Mingdao CHEN
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
Objective To observe expression of resistin gene in adipose tissue of SD rats fed with high-fat diet and its correlation to insulin resistance, diabetes mellitus and leptin. Methods Using RT-PCR to mRNA from adipose tissue, the products were obtained, purified, constructed and squenced. Once its squence was verified, semi-quantitative RT-PCR and Northern Blot were employed to study its expressions in adipose tissues of rats with different diets. Oral glucose tolerance test and insulin release test were performed in 2 groups of rats. Results (1) The resistin gene was successfully constructed and sequenced, which was in accordance with the data of genbank. (2) Compared with the control, the expression of resistin in adipose tissue of high-fat diet rats was significantly enhanced, and blood glucose, serum insulin and leptin levels were also elevated in high-fat diet rats (P
5.Effects of gefitinib on embryonic development and expression of tumor resistance genes in zebrafish
Rongying HU ; Liping SHU ; Zhixu HE ; Congjie SUN ; Zhicao LI ; Lu JIN
Chongqing Medicine 2017;46(32):4469-4472
Objective To investigate the effects of gefitinib on early embryonic development and expression of abcb4 tumor resistance genes in zebrafish.Methods Zebrafish were adopted as experimental animals and divided into gefitinib group,mixture of doxorubicin and gefitinib group and blank group.Zebrafish embryos of 0.5-1.5 hours after fertilization(0.5-1.5 hpf) were exposed to different concentrations of gefitinib,and then embryo development of zebrafish in 24-120 hpf was observed and the number of death,hatch and malformation was recorded.The embryo mortality was calculated under different concentrations of gefitinib at different time points,and the numerical value of IC50 was calculated;Hatching rates of zebrafish embryo in 48 hpf and 72 hpf and malformation rates of zebrafish embryo in 120 hpf were calculated.The zebrafish embryos exposed to different concentrations of gefitinib in different groups were collected,and the expressionof abcb4 gene in zebrafish embryos was detected by real-time quantitative PCR.Results Gefitinib IC50 in zebrafish embryos was 16.18 μmol/L.Compared with the control group,higher dosage (20 μmol/L) of gefitinib in other two groups significantly decreased hatching rates of embryos,and had obvious embryonic lethal effects and teratogenic effects.Moreover,the mRNA levels of the abcb4 gene in the zebrafish embryos of gefitinib group were not significantly changed,whereas the mRNA levels of the abcb4 gene in mixture of doxorubicin and gefitinib group were significantly different(P<0.05).Conclusion Gefitinib has no significant effects on the expression of abcb4 tumor resistance gene in early development of zebrafish embryos(P>0.05),but it can reverse the drug resistant effects of doxorubicin,suggesting that zebrafish can construct tumor resistance model.