1.Effect of estradiol on proliferation of rat hippocampal neural stem cells
Xiaodong LIU ; Xianan ZHANG ; Ning HAO ; Qianqian JU ; Jianbing QIN ; Meiling TIAN ; Guohua JIN
Acta Anatomica Sinica 2014;(5):627-632
Objective The aim is to observe the role and mechanism of estradiol ( E2 ) on the proliferation of rat hippocampal neural stem cells ( NSCs ) .Methods Twenty hippocampi from embryonic 17-day ( E17 ) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol .The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay .The expression of estrogen receptors ( ERαand ERβ) was measured by immunofluorescence staining combined with Nestin double labeling . Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10 -10 to 10 -8 mol/L.The number of cell proliferation and the viability of cells were best at the concentration of 10 -8 mol/L compared to the other groups .However, when the estradiol concentration was increased from 10-8 to 10 -6 mol/L, the cell proliferative capacity declined gradually .Double immunofluorescence labeling showed that the two types of estrogen receptors ( ERαand ERβ) were expressed in the cultured hippocampal NSCs .Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range , and ERαand ERβmay be involved in the estradiol-induced proliferation .
2.Effects of sirolimus and starved culture on autophagy of human A431 cells
Yun ZHANG ; Xiaowen YANG ; Qianqian XU ; Xin SHI ; Xu CHEN ; Li LI ; Song XU ; Dan HUANG ; Mei JU ; Kun CHEN ; Heng GU
Chinese Journal of Dermatology 2016;49(11):776-780
Objective To evaluate effects of sirolimus(a classic autophagy inducer)and starved culture on autophagy of a human cutaneous squamous cell carcinoma cell line A431. Methods Cultured A431 cells and HeLa (a human cervical carcinoma cell line)cells were both classified into 5 groups to be treated with DMEM alone(control group), 0.1%dimethyl sulfoxide alone(DMSO group), 20 nmol/L sirolimus(20?nmol/L sirolimus group), 80 nmol/L sirolimus(80?nmol/L sirolimus group), and Earle′s balanced salt solution(EBSS group)respectively. After 4?hour treatment, Western blot analysis was performed to measure the expressions of autophagy?related markers microtubule?associated protein 1 light chain 3A/3B (LC3A/B) and recombinant gamma?aminobutyric acid receptor associated protein(GABARAP), and acridine orange staining to determine autophagy levels in these cells. Results As Western blot analysis showed, the ratio of LC3A/B?Ⅱto LC3A/B?Ⅰin A431 cells was similar between the control group and DMSO group(P > 0.05), but significantly higher in the 20?nmol/L sirolimus group, 80?nmol/L sirolimus group and EBSS group than in the control group(all P < 0.05). Western blot results from HeLa cells were similar to those from A431 cells. Bivariate correlation analysis revealed that the protein expression of GABARAP was positively correlated with that of LC3A/B ?Ⅰ in both HeLa cells(r = 0.869, 95% CI: 0.807 - 0.999, P = 0.051)and A431 cells(r = 0.837, 95% CI: -0.173 - 0.989, P = 0.037), but negatively correlated with that of LC3A/B?Ⅱ in both HeLa cells(r = -0.742, 95% CI: -0.982 - 0.406, P = 0.042)and A431 cells(r = - 0.684, 95% CI: -0.977 - 0.500, P = 0.047). Acridine orange staining showed that the percentages of autophagosome?positive A431 cells and HeLa cells were significantly increased in both the 80?nmol/L sirolimus group(23.750% ± 0.260% and 33.307% ± 0.715% respectively)and EBSS group(32.450% ± 0.488% and 66.097% ± 1.141% respectively) compared with the control group(15.987% ± 0.242% and 14.117% ± 0.295%, respectively, all P < 0.05). Conclusion The classic autophagy inducer sirolimus and starved culture can upregulate the autophagy level of A431 cells, and GABARAP may be highly correlated with LC3A/B.
3.Color Doppler ultrasound visualizes early post-traumatic heterotopic ossification in animal model
Qianqian HE ; Xiaoxue CHEN ; Shaoling YANG ; Jinyong JU ; Haijun XIAO ; Cong WANG ; Kun ZHAO ; Jing HU
Chinese Journal of Ultrasonography 2020;29(2):175-179
Objective:To validate the role of color Doppler ultrasound in an animal model to detect early heterotopic ossification (HO) after brain-traumatic/burn/tenotomy.Methods:Forty-four rats were randomly divided into two groups. Rats in experimental group ( n=22) were operated to build brain-traumatic/burn/tenotomy model and others in control group ( n=22) underwent only skin incision injury. Color Doppler ultrasound, X-ray film examination at 2, 3, 4, 6, 8 and 10 weeks post-injury were performed to follow up the progression of HO in both groups respectively. Histology was used to confirm bone formation. Results:In the experimental group, disorder structure with a hypoechoiccore in treated Achilles tendon was visualized using color Doppler ultrasound in the 2nd week. Additional tiny hyperechoic foci were observed in the 3rd week, which increased in the fourth week and fused into a mineralized island in the sixth week. No obvious abnormality was found in control group at the aforementioned time point. X-ray could detect heterotopic bone tissue in the sixth week in the experimental group but not in the control group. X-ray and HE stainning had confirmed bone formation in the tenth week in the experimental group.Conclusions:Color Doppler ultrasound can detect early HO and continuously follow up the progression of HO.
4.Therapeutic effects of enteral nutrition support combined with rehabilitation therapy in senile dementia patients with malnutrition
Qianqian TU ; Xiping TUO ; Wenjun ZHANG ; Haitao CHEN ; Ning ZHAO ; Zeya JU ; Rihan WU ; Feng HUANG
Chinese Journal of Geriatrics 2019;38(3):274-277
Objective To investigate changes in nutritional status in senile dementia patients with malnutrition treated with enteral nutrition combined with rehabilitation therapy evaluated by using the Mini Nutritional Assessment-Short Form(MNA-SF).Methods Fifty hospitalized patients with senile dementia and malnutrition at the internal medicine department of our hospital from July 2014 to July 2017 were enrolled and further divided into an enteral nutrition group(a control group,n =25)and an enteral nutrition combined with rehabilitation therapy group(an observation group,n=25).Anthropometric parameters including body mass index,biceps circumference,calf circumference,triceps skin-fold thickness,and blood biochemical parameters including hemoglobin,total cholesterol,triacylglycerol,albumin,lymphocyte count before and after treatment were compared between the groups.Results Anthropometric parameters such as body mass index,biceps circumference,calf circumference and triceps skin-fold thickness had no significant difference between the two groups before treatment (P > 0.05),while after 30 days of treatment,anthropometric parameters saw improvement in both groups compared with those before treatment,and the improvement was greater in the observation group than in the control group(P <0.01).Blood biochemical parameters had no significant difference between the two groups before treatment(P>0.05).After 30 days of treatment,levels of total cholesterol and triacylglycerol had no significant difference compared with those before treatment or between the two groups(P >0.05),while levels of hemoglobin,albumin and lymphocyte count improved compared with those before treatment (t =2.645,2.843,2.967,respectively,P <0.01),and had significant differences between the groups (t =2.548,2.864,2.976,respectively,P<0.01).Conclusions Enteral nutrition combined with rehabilitation therapy has a measurable,positive influence on nutritional status in senile dementia patients with malnutrition.