1.Effect of different dose of GM1 on inducing human adipose tissue-derived stromal cells into neural cells
Bin LIU ; Menghai WU ; Jinxia ZHANG
Journal of Clinical Neurology 1992;0(01):-
Objective To investigate the effect of different dose of GM1 on inducing human adipose tissue-derived stromal cells (ADSCs) into neural cells. Methods Human ADSCs were isolated from human liposuction tissues and cultured in NIM with different dose of GM1, which was used to induce ADSCs into neural cells. The experiment was separated into six groups: control group (?-MEM), Group A (NIM), group B (NIM+50 ?mol/L GM1), group C (NIM+100 ?mol/L GM1), group D (NIM+200 ?mol/L GM1) and group E (NIM+400 ?mol/L GM1). Cells' growth and morphologic changes were observed under invert phase-contrast microscopy. The expressions of Nestin, NSE, MAP2 and GFAP were identified by immunocytochemistry methods.Results (1)ADSCs could differentiate into neural cells in NIM with different dose of GM1. The number of neural cells differentiated from ADSCs increased in a certain period, especially in group E at each time point (all P
2.Ca~(2+) signaling mediated salidrosides promotes directional differentiation of mouse bone marrow mesenchymal stem cells into nerve cells
Jingjing PEI ; Run WU ; Hongbin ZHAO ; Xudong LIU ; Jun HU ; Menghai BAI
Chinese Journal of Tissue Engineering Research 2010;14(10):1808-1812
BACKGROUND:Tranditional Chinese medicine,which possesses anti-oxidation properties,can promote directional differentiation of mouse bone marrow mesenchymal stem cells(BMMSCs)into nerve cells.Salidrosides,as the effective constituent of Rhodiola,have strong anti-oxidation function.OBJECTIVE:To investigate the molecule mechanism of salidrosides induced differentiation of mouse BMMSCs into nerve cells.METHODS:When in vitro cultured BMMSCs reached 80% confluency,the cells were assigned into 3 groups.Cells in the control group were cultured by complete culture medium;those in the induction and positive control groups were cultured by complete culture medium adding 20 mg/L salidrosides or 0.1 mg/L nerve growth factors(NGF).The related gene and protein of nerve cells were detected using RT-PCR and Western blot method at 12 hours after culture.After that,the cells in the induction group were divided into 3 groups,the blocking agents EGTA(Ca~(2+) chelator),Nifedipine(L-type Ca~(2+) channel blocker)and LY294002(IP3 receptor blocking pharmacon)were applied to block the cellular Ca2* signal pathway respectively for 12 hours.RT-PCR and Western blot methods were used to study the signal transduction of the salidrosides.RESULTS AND CONCLUSION:①The expression of neuron specific enolase(NSE),B-Tubulin III,Nurri mRNA could be found ir the induction and positive control groups,instead of the control group;The expression abundance of the positive control group was smaller than that of the induction group.The expression abundance of GFAP mRNA was very low in each group,but the c-fos mRNA was expressed abundantly in the induction group.②Compared with the positive control group,the induction group could promote the NSE expression obviously,which was no expressed in the blank control group.?The expression of NSE and Nurri were conspicuously down-regulated when the Ecto Ca~(2+) and L-type Ca~(2+) channel and IP3 receptor were blocked respectively.Salidrosides can induce the differentiation BMMSCs into nerve cells. Ca~(2+) signaling and IP3 dependent Ca~(2+) signaling pathway play an important role in transduction salidrosides signal in BMMSCs differentiation.
3.Neuronal differentiation of adipose tissue-derived stromal cells
Bin LIU ; Menghai WU ; Jing DONG ; Ning LIU ; Jianmin LI ; Jinxia ZHANG ; Shiying LI ; Ruimin WANG ; Guiliang CHEN
Chinese Journal of Tissue Engineering Research 2010;14(1):15-18
BACKGROUND: Appropriate seed cell is important for transplantation in the treatment of cerebrovascular disease and other central nervous system disease.OBJECTIVE: To investigate the capacity of human adipose tissue-derived stromal cells (ADSCs) to differentiate into neurons.METHODS: The fatty tissue was harvested from removed abdominal unnecessary fat of healthy adult with no communicable disease or endocrine disease. Human ADSCs were isolated from human liposuction tissues and cultured in neural induction medium with GM1. Invert phase-contrast microscopy was used to observe morphology changes of ADSCs. The expression of nestin, neuron specific enolase (NSE) and microtubule-associated protein 2 (MAP2) were identified by immunocytochemistry.RESULTS AND CONCLUSION: The majority of cells displayed typical appearance of neuronal-like cells following induction. Following 1 hours of induction, some cells began to express nestin, and NSE and MAP2-positive cells were observed at 5 hours. ADSCs can differentiate into neurons, and the differentiated neurons have the capacity of expressing nestin, NSE and MAP2.
4.Expression of insulin-like growth factor binding protein 3 in cerebral infarction rats and its involvement in Smad signaling pathway
Cui ZHANG ; Menghai WU ; Yunyan DUO
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(3):331-335
Objective To investigate the role of Smad signaling pathway in rat model of cerebral in-farction and explore the expression of insulin-like growth factor binding protein 3(IGFBP-3)in brain tissue and its relationship with neural function.Methods Sixty healthy adult male SD rats were randomly and equally divided into model group,sham-operation group,and normal control group.The model of cerebral infarction was established by using intraluminal thread occlusion,and the rats of the sham-operation group were only given exposure of the internal carotid artery and direct suture of the incision.In 1 week after successful modeling,Modified Neurological Seve-rity Score(mNSS)was used to evaluate the neurological function.HE staining was employed to observe the histopathological changes in the brain tissues.Western blotting and RT-PCR were adopted to detect the brain expression of IGFBP-3,Smad2 and Smad4 at protein and mRNA le-vels.Spearman correlation analysis was conducted to analyze the correlation among the expression levels of IGFBP-3,Smad2,Smad4 and P21.Results HE staining displayed that obvious brain ede-ma,characterized by disordered arrangement of brain cells,increased microglia,and blurred nucleo-lus of brain cells were observed in the rats of the model group,with the area of cerebral infarct of 20.55%.The mNSS score and the protein and mRNA levels of IGFBP-3,Smad2 and Smad4 were significantly higher,but the P21 protein and mRNA levels were obviously reduced in the model group than the sham-operation group and normal control group(P<0.05,P<0.01).Spearman correlation analysis showed that the mRNA level of IGFBP-3 in cerebral infarction rats was posi-tively correlated with the mNSS score and mRNA expression levels of Smad2 and Smad4(r=0.568,r=0.623,r=0.597;P<0.01),and negatively with P21 mRNA level in the brain tissue(r=-0.573;P<0.01).Conclusion The level of IGFBP-3 is significantly increased in brain tissue of rats with cerebral infarction,and it is closely associated with neural function of these rats,which may be related to Smad signaling pathway.
5.Establishment of screening models for nonalcoholic fatty liver disease in the adult Blang population
Yebei LIANG ; Chunguang YANG ; Huadong ZENG ; Ruwei TAO ; Qiuming HU ; Xiaoying TANG ; Huaxiang SHI ; Wei WU ; Xuhong HOU ; Weiping JIA
Journal of Clinical Hepatology 2021;37(12):2861-2868
Objective To establish simple screening models for nonalcoholic fatty liver disease (NAFLD) in the adult Blang population. Methods Based on the survey data of metabolic diseases in the Blang people aged 18 years or above in 2017, 2993 respondents were stratified by sex and age (at an interval of 5 years) and then randomly divided into modeling group with 1497 respondents and validation group with 1496 respondents. Related information was collected, including demographic data, smoking, drinking, family history of diseases and personal medical history, body height, body weight, waist circumference, and blood pressure, and related markers were measured, including fasting plasma glucose, 2-hour postprandial plasma glucose or blood glucose at 2 hours after glucose loading, triglyceride, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transpeptidase. The chi-square test was used for comparison of categorical data between two groups. Logistic regression analysis was used to establish the screening model. The area under the receiver operating characteristic curve (AUC), sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value were used to evaluate the screening performance of established models versus existing models in the study population, and the DeLong method was used for comparison of AUC. Results Three screening models for NAFLD were established based on physical and biochemical measurements, i.e., simple noninvasive model 1 (age, body mass index, and waist circumference), noninvasive model 2 with the addition of blood pressure, and model 3 with the combination of hematological parameters (diabetes and ALT/AST). In the modeling group, the three models had an AUC of 0.881 (95% confidence interval [ CI ]: 0.864-0.897), 0.892 (95% CI : 0.875-0.907), and 0.894 (95% CI : 0.877-0.909), respectively, and there was a significant difference between model 1 and models 2/3 ( P =0.004 0 and P < 0.001); in the validation group, the three models had an AUC of 0.891 (95% CI : 0.874-0.906), 0.892 (95% CI : 0.875-0.907), and 0.893 (95% CI : 0.876-0.908), respectively, and there was no significant difference between the three groups ( P > 0.05). Based on the overall consideration of screening performance, invasiveness, and cost, the simple noninvasive model 1 was considered the optimal screening model for NAFLD in this population. Model 1 had the highest Youden index at the cut-off value of 5 points, and when the score of ≥5 points was selected as the criteria for NAFLD, the model had a sensitivity of 86.5%, a specificity of 79.7%, a positive predictive value of 50.3%, and a negative predictive value of 96.1% in the modeling group and a sensitivity of 85.6%, a specificity of 80.6%, a positive predictive value of 51.7%, and a negative predictive value of 95.8% in the validation group. Conclusion The NAFLD screening models established for the adult Blang population based on age and obesity indicators have relatively higher sensitivity, specificity, and negative predictive value, and this tool is of important practical significance for the intervention of NAFLD and its closely related metabolic diseases in this population.