1.Isolation, culture, identification and labeling of neural stem cells from newborn guinea pigs
Ying WANG ; Mingmin DONG ; Ying DU
Chinese Journal of Tissue Engineering Research 2006;10(29):157-159,插5
BACKGROUND: Transplantation of neural stem cells into inner ear brings hope for treatment of sensorineural deafness, while guinea pig is the preferred animal in developing animal models of aminoglycosides poisoning-induced deafness. Successful animal models have been established after study for several scores of years. How to isolate neural stem cells and label them from hippocampi of guinea pigs is one of the problems to be solved in transplantation of stem cells into inner ear.OBJECTIVE: To isolate and culture neural stem cells from hippocampi of newborn guinea pigs so as to lay basis for transplantation of neural stem cell in the treatment of sensorineural deafness.DESIGN: Single sample observation SETTING: Staff Room of Microbiology and Immunology, Zhengzhou University MATERIALS: This experiment was carried out at the Staff Room of Microbiology and Immunology, Zhengzhou University from March to June 2005. Newbornguinea pigs born within 24 hours of either gender, weighing 50 to 80 g, provided by Experimental Animal Center of Zhengzhou University were used in this experiment.METHODS: Neural stem cells from the hippocampal tissue of newborn guinea pigs were isolated and cultured by serum-free medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF).Expression of nidogen was detected with immunocytochemical staining.Neural stem cells were labeled by DAPI.MAIN OUTCOME MEASURES: ① In vitro growth of neural stem cells; ② neural stem cells nidogen expression was identified by immunofluorescent staining; ③ labeled stem cells by fluorescence and detected the labeling rate.RESULTS: ①Clone-like cluster of neural stem cells were obtained from hippocampal tissue ② Neural stem cells could express nidogen. ③Labeling rate of DAPI was 93.4% and no attenuation of fluorescence brightness as observed after 8 generations in culture.CONCLUSION: The cells obtained from hippocampal tissues of newborn guinea pigs can be used as donor cells in the treatment of neurosensory deafness by transplanting neural stem cells, for their obvious proliferation ability and high-labeled efficiency with fluorescent dye.
2.Clinical observation of Zishen Ningxin capsule combined with telmisartan in the treatment of perimenopausal hypertension
Bei WANG ; Mingmin DU ; Chengyi WANG ; Da FU ; Jing ZHANG ; Qiang WANG
Chinese Journal of Pharmacoepidemiology 2024;33(1):52-59
Objective To observe the efficacy of Zishen Ningxin capsule combined with telmisartan in the treatment of perimenopausal hypertension,and its influence on the patients'blood pressure,biochemical indexes and TCM symptom scores.Methods 300 patients with perimenopausal hypertension were randomly divided into control group and observation group with 150 patients in each group.The control group was given telmisartan treatment,and the observation group was given Zishen Ningxin capsule on the basis of the control group.The treatment course of both groups was 8 weeks.The dynamic blood pressure and coefficient of variation of blood pressure at 24 h before and after treatment were observed and compared between two groups(24 h systolic blood pressure and coefficient of variation,24 h diastolic blood pressure and its coefficient of variation),sex hormone levels(serum estradiol,follicle stimulating hormone,testosterone,progesterone),blood lipid levels(triacylglycerol,total cholesterol,low density lipoprotein cholesterol,high density lipoprotein cholesterol),renin and hypersensitive C-reactive protein levels,as well as the changes of TCM symptom score were evaluated for the efficacy of the two groups.Results After treatment,the 24 h systolic blood pressure and its coefficient of variation,24 h diastolic blood pressure and its coefficient of variation,the levels of sex hormones,blood lipids,renin and hypersensitive C-reactive protein,as well as TCM symptom scores in two groups were significantly improved(P<0.05),and all indexes in the observation group were better than those in the control group(P<0.05).The total effective rate of observation group was significantly higher than that of control group(P<0.05).There was no significant difference of the incidence of adverse reactions in the two groups(P>0.05).Conclusion Zishen Ningxin capsule combined with telmisartan has significant clinical efficacy in the treatment of perimenopausal hypertension,which can effectively reduce blood pressure,relieve symptoms,improve the levels of sex hormones,blood lipids,renin and hypersensitive C-reactive protein.
3.Mechanism of Xiezhuo Jiedu Recipe Regulating Ferroptosis in Treatment of Ulcerative Colitis Based on Bioinformatics and Animal Experiments
Chaodi SUN ; Jianping LIU ; Mingmin DU ; Xin KANG ; Jiancong CUI ; Yuan ZHAO ; Sujie JIA ; Xiaomeng LANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):166-173
ObjectiveThe bioinformatics method was used to screen ferroptosis differential genes (FRGs) closely related to ulcerative colitis (UC), and animal experiments were conducted to verify whether the mechanism of Xiezhuo Jiedu recipe in treating UC is related to the regulation of ferroptosis. MethodThe differentially expressed genes (DEGs) of colonic mucosa tissue of UC patients were obtained from the GEO database, and the intersection of the genes with ferroptosis genes was used to obtain FRGs. The core FRGs were obtained by cluster analysis, minimum absolute contraction and selection operator (LASSO) regression, and receiver operating characteristic curve (ROC) curve analysis. In animal experiments, the UC mouse model was prepared by making the mouse freely drink 2.5% dextran sodium sulfate (DSS). Xiezhuo Jiedu recipe and mesalazine were given by gavage for seven days, and the inflammatory infiltration of colonic mucosa was observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of E3 ubiquitin ligase (FBXW7), zinc finger protein (ZFP36), solute carrier family 7 member 11 (SLC7A11), and Toll-like receptor 4 (TLR4) in colon tissue. The protein expression levels of FBXW7, ZFP36, SLC7A11, and TLR4 in colon tissue were detected by Western blot. ResultDataset GSE87466 was screened from the GEO database, and its intersections with the ferroptosis gene were analyzed to obtain 21 FRGs. After cluster analysis, LASSO regression, and ROC analysis, core FRGs (FBXW7, ZFP36, SLC7A11, and TLR4) were obtained. Immunoinfiltration analysis showed significant differences in the expression of initial B cells, M1 macrophages, plasma cells, and M2 macrophages in the colonic mucosa tissue of UC mice, and there was a significant correlation between core FRGs and these immune cells. Further animal experiments showed that the colonic mucosa tissue of mice in the model group was disorganized and infiltrated by a large number of inflammatory cells. The inflammation of the colonic mucosa tissue of mice in each group was relieved to varying degrees after treatment with Xiezhuo Jiedu recipe and mesalazine, while the colonic mucosa tissue of mice in the high-dose group of Xiezhuo Jiedu recipe showed almost no inflammatory changes. Compared with the normal group, the protein and mRNA expressions of FBXW7, ZFP36, SLC7A11, and TLR4 in the model group were significantly increased, and the expression of core FRGs in colonic mucosa tissue of mice in all groups was significantly down-regulated after treatment with Xiezhuo Jiedu recipe and mesalazine. ConclusionFBXW7, ZFP36, SLC7A11, and TLR4 are ferroptosis genes closely related to the pathogenesis of UC, and Xiezhuo Jiedu recipe can significantly alleviate colonic mucosa inflammation in mice by down-regulating core ferroptosis genes.