1. The system for directional and stable differentiation of amniotic fluid stem cells into neuron-like cells
Chinese Journal of Tissue Engineering Research 2020;24(13):2055-2060
BACKGROUND: Neuronal regeneration using stem cell differentiation has gained a lot of attentions from researchers. Although embryonic stem cells and induced pluripotent stem cells have good potential for neuronal differentiation, a high risk of tumor development in vivo limits the further study. OBJECTIVE: To establish a stable system for sorting, culture and neuronal differentiation of amniotic fluid stem cells, and to explore the feasibility as seed cells for neuronal regeneration. METHODS: Amniotic fluid sample (10 mL) was obtained at 19-22 weeks of pregnancy under B-ultrasound guidance, and amniotic fluid stem cells were isolated by c-Kit magnetic beads. The markers Oct-4 and Sox2 of amniotic fluid stem cells were identified by immunofluorescence. The expression levels of c-Kit, Oct-4, Sox2 and Nestin in amniotic fluid stem cells after multiple passages were detected by RT-PCR. Then, the cells were cultured by hanging drop for 4 days to observe the embryoid bodies-like structures. Amniotic fluid stem cells were induced to differentiate into neurons using two-stage method. The expression levels of Neuro D and Tuj1 were observed by immunofluorescence. RESULTS AND CONCLUSION: (1) About 1% of amniotic fluid stem cells were positive for c-Kit. (2) (75.0±4.6)% of amniotic fluid stem cells expressed Oct-4 and (86.0±2.8)% of the cells expressed Sox2. (3) The expression levels of c-Kit, Oct-4, Sox2 and Nestin detected by RT-PCR did not change with passage times. (4) Embryoid bodies-like structures formed after hanging drop culture. (5) Immunofluorescence results showed that amniotic fluid stem cells expressed neuronal marker Tuj1, but without the typical morphological features. RT-PCR detected the expression of Tuj1 in different amniotic fluid stem cell specimens as well as in the same sample after several passages. (6) Amniotic fluid stem cells could have the characteristics of neuron-like cells after induction with basic fibroblast growth factor, brain-derived neurotrophic factor, and neurotrophin factor 3 in two stages, and could express neural stem cell marker Neuro D and neuronal marker Tuj1.
2. Establishment and verification of cytochrome P450 3 A enzyme induction model in female rats
Chinese Pharmaceutical Journal 2013;48(12):995-999
OBJECTIVE: To establish a female rat CYP3A induction model which is used for studying CYP3A-mediated drug-drug interactions. METHODS: Female SD rats which were fed with standard diet were randomly divided into two groups, one was the control group, and the other one was the experimental group. The rats in the experimental group were administered respectively 20, 50, 80, 100, and 150 mg·g-1d-1 dexamethasone by gavage for 3 d to induce CYP3A enzymes. 24 h after 3 d, liver tissues were taken from both groups of rats and rat liver microsomes were prepared. CYP3A4 activity was determined with testosterone as the probe substrate. RESULTS: Testosterone metabolic rate was 31.68% in blank liver microsomes, and were 40.64%, 61.36%, 82.44%, 85.8%, and 83.36% in dexamethasone-induction group. Testosterone metabolic rate was improved by up to 160.23% after dexamethasone induction at 80 mg·g-1d-1 in female rats. CONCLUSION: Dexamethasone induction at 80 mg·g-1d-1 can significantly increase liver CYP3A enzyme activity in female SD rats, and the model can be used to study CYP3A-mediated drug-drug interactions.
3.Current prevalence and correlates of depressive disorder among outpatients with essential hypertension from cardiovascular divisions of tertiary general hospitals
Xianguo ZHANG ; Qiang FAN ; Qiang FANG ; Juzhen JING ; Ling ZONG
Chinese Journal of Nervous and Mental Diseases 2015;(4):193-198,223
Objective To investigate the current prevalence of depressive disorder and the associated factors in pa?tients with essential hypertensive from cardiovascular divisions of tertiary general hospitals. Methods A Chinese version of Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) Axis I Disor?ders was conducted by five mental health professionals on 876 essential hypertensive outpatients at cardiovascular divi?sions of tertiary general hospital. Data on the socio-demographic and clinical characteristics of these hypertensive pa?tients were also collected. Results The current prevalence of depressive disorders among essential hypertensive outpa?tients of tertiary hospitals was 16.6%(145/876). Risk factors for depressive disorders in hypertensive patients included fe?male gender (OR=2.817, 95%CI:1.161~6.885), being 65-year-old and older (OR=1.747, 95%CI:1.118~2.688), non-marital status (OR=2.023, 95%CI:1.203~3.398), gradeⅢhypertension (OR=1.776, 95%CI:1.226~2.333) and car?diovascular diseases (OR=2.018, 95%CI:1.427~2.587). Conclusion Essential hypertensive outpatients, who receive med?ical services at cardiovascular divisions of tertiary general hospital, have high prevalence of depressive disorders. Clini?cians should pay more attention to the mental health of outpatients with essential hypertension.
4.Meta-analysis of randomized controlled trials of sodium hyaluronate eye drops treating dry eye
Hong-ling, WEI ; Zhan-zong, LI ; Ya-fang, LI
Chinese Journal of Experimental Ophthalmology 2013;(5):496-500
Background Some researching results of sodium hyaluronate eye drops treating dry eye have been reported home and abroad.To evaluate these results using the method of evidence-based medicine (EBM) has an important clinical significance.Objective This secondary study was to systematically evaluate the efficacy of sodium hyaluronate eye drops for the treatment of dry eye.Methods Meta analysis was adopted in this study.The electronic bibliographic databases were searched according to the prescribed strategy,including Medline,PubMed,VIP,CBM,CNKI,Wanfang Data using the method of Cochrane systematic review.The literature published time was limited from the commencement of each database to December,2011.Randomized controlled trials (RCTs) of sodium hyaluronate eye drops treating dry eye were included and the methodological quality of the RCTs was assessed by two evaluators using Jadad measuring scale,and homogeneous evaluation by Meta-analysis was performed.Statistical analysis of clinical data was performed using RevMan 5.0 software provided by the Cochrane Collaboration.With odds ratio (OR) value as the effect index,the randomized effect model was used to analyze the clinical heterogeneity.Results A total of 4 RCTs involving 686 patients were included.Jadad scale evaluation showed 7 points in 2 literature,4 points in 1 literature and 2 points in 1 literature.In all the RCTs,344 patients with dry eye were treated using sodium hyaluronate as the sodium hyaluronate eye drops group,and 342 dry eyes applied placebo,normal saline solution,hydroxymethyl cellulose,hydroxypropyl methyl cellulose respectively as the control group.Meta-analysis indicated that the total effective rate (OR) was 2.55 with the 95% confidence interval (CI) 0.94-6.93.The symptoms were improved in 183 cases in the experiment group and 130 cases in the control group.No significant difference was found in the OR between the sodium hyaluronate eye drops group and the control group (Z =1.84,P=0.07).Conclusions Meta-analysis indicates that sodium hyaluronate eye drops can improve the symptom of dry eye,but have no significence with the control group.However,owing to fewer number and uneven quality of included RCTs,the large sample size and multicenter of RCTs is still needed to verify the superiority of sodium hyaluronate eye drops for dry eye.
5.Internal rib fixation combined with external chest fixation and mechanical ventilation for traumatic flail chest with pulmonary contusion
Ziteng ZHANG ; Zhiliang HU ; Ling ZONG ; Ming MA ; Chunyang MENG
Chinese Journal of Trauma 2015;31(10):895-898
Objective To analyze the clinical efficacy of internal rib fixation combined with external chest fixation and mechanical ventilation for traumatic flail chest with pulmonary contusion.Methods Sixty cases of traumatic flail chest with pulmonary contusion treated from January 2011 to December 2014 were assigned to experimental group (30 cases) and control group (30 cases) according to the random number table.Patients in control group received thoracic external fixation combined with mechanical ventilation.In experimental group the patients received the same care but in addition they had rib fixation.The two groups were compared with regard to general condition, arterial blood gas as well as vital signs before treatment and 24 hours after treatment, lung function 3 months after discharge, complication rate and mortality.Results ICU stay [(6.8 ± 1.0) d], hospital stay [(15.0 ± 1.8) d] and duration of mechanical ventilation [(4.8 ± 1.0) d] in experimental group were significantly lower than these in control group [(13.6 ± 2.5) d, (21.4 ± 2.6) d, (10.3 ± 1.3) d, respectively] (P < 0.01).After treatment for 24 hours in either group, the pH value, PaO2, oxygenation index and systolic pressure (SBP) were significantly increased, and partial pressure of carbon dioxide (PaCO2), heart rate (HR) and respiratory rate (RR) were significantly lowered as compared to these before treatment (P <0.05).After treatment for 24 hours, experimental group demonstrated significantly enhanced levels in pH value,PaO2, oxygenation index and SBP but lowered levels in PaCO2, HR and RR as compared to control group (P < 0.05).Three months after discharge, forced vital capacity (FVC), forced expiratory volume in one second (FEV1), peak expiratory flow (PEF), 75% forced expiratory flow (FEF75%) and total lung capacity (TLC) in experimental group were (81.7 ±2.6)%, (75.4 ±4.1)%, (83.2 ±4.6)%,(69.1 ±2.3)%, and (88.7 ±3.4)% respectively, significantly higher than (69.0 ±3.6)%, (71.3 ± 3.9) %, (78.9 ± 4.3) %, (62.3 ± 3.3) %, and (79.0 ± 4.6) % respectively in control group (P <0.01).In experimental group, there were four cases of pulmonary infection (13%), three pulmonary atelectasis(10%) and one intercostal neuralgia (3%).In control group, there were six cases of pulmonary infection (20%), five pulmonary atelectasis (17%), four intercostal neuralgia (13%) and four thoracocyllosis(13%)(P <0.05).There was no significant difference in mortality between the two groups.Conclusion Treatment effect of internal rib fixation for flail chest with pulmonary contusion is improved in combination with mechanical ventilation and external chest fixation.
6.Current studies on assessment of scientific research capability in domestic and foreign medical institutions:A review
Ling YI ; Weitao ZONG ; Lijuan LI ; Changqin LI ; Lei XU
Chinese Journal of Medical Library and Information Science 2015;(4):15-20
After the systems, methods and criteria for assessment of scientific research capability in USA, Germany, Britain and Japan were described, the advances in studies on assessment of scientific research capability in China were summarized, their characteristics were compared, and the enlightenments to perform assessment of medical research institution capability were elaborated.
7.Case of Sjogren syndrome.
Yin-Ping WANG ; Qing ling WEI ; Zong-Quan ZHANG
Chinese Acupuncture & Moxibustion 2013;33(10):918-918
8.Effects of carbon nanoparticles-epirubicin suspension on cell proliferation and apoptosis in breast cancer MCF-7 cells
Yongsong ZHANG ; Quankun LIANG ; Ling ZHONG ; Xiaowei QI ; Beige ZONG ; Junze DU ; Yi ZHANG ; Linjun FAN
Chinese Journal of Endocrine Surgery 2017;11(2):110-114
Objective To explore the effect of carbon nanoparticles-epirubicin suspension on proliferation and apoptosis in human breast cancer MCF-7 cells.Methods MCF-7 cells were cultured with different concentrations of CNP-EPI in vitro.CCK-8 assay was used for determinate inhibition effect of CNP-EPI on the proliferation of MCF-7 cells at different concentration and different time.According to the determination of IC90,5 μg/ml CNP-EPI was selected,and cell morphology and cell apoptosis rates were observed after 24 h.Results The inhibition effect of the CNP-EPI was stronger significantly in CNP-EPI group than in normal control group within 24 h,48 h,72 h when the concentration was from 1 μg/ml to 200 μg/ml (P<0.01).The inhibition of CNP-EPI on the proliferation of MCF-7 cells was gradually strengthened in a dose-dependent relation within the same time,and the inhibition effect is reduced in the same concentration of drugs with the time extension,but it still has a strong inhibitory effect in 72 h,and the inhibition effect of different concentration of CNP was not obvious on MCF-7 cells.Obvious changes of cell morphology were observed under inverted microscope such as:a lot of dead cells,cell adherent growth poor,cell morphology became round and karyopycnosis etc,in 5 μg/ml CNP-EPI group after 24 h.However,no obvious abnormity of cell norphology was observed in normal control group and corresponding CNP group.Late apoptosis rate was (14.57±2.41) %,the mortality rate could reach (78.63±-20.55)% in 5 μg/ml CNP-EPI group after 24 h.The mortality rate and apoptosis rate of cells was higher significantly in CNP-EPI group than in CNP group and normal control group (P<0.05).Conclusion CNP-EPI can obviously inhibit the proliferation or kill human breast cancer MCF-7cells,and the inhibition effect of CNP-EPI on proliferation of breast cancer cells might be the result of delayed releasing of EPI.
9.Effects of Mycobacterium vaccae vaccine on immunological function in elderly patients with stable chronic obstructive pulmonary disease
Zong-xiao SHANGGUAN ; Ren-guo ZHAO ; Ling-yun LIU ; Feng GONG ; Xiao CI ; Ming-hua ZHANG ;
Chinese Journal of Geriatrics 2010;29(5):382-385
Objective To investigate the effects of Mycobacterium vaccae vaccine on immunological function and clinical character in elderly patients with stable chronic obstructive pulmonary disease (COPD). MethodsA total of 100 elderly patients with stable COPD were randomly divided into immunotherapy group (group A, n= 50) and non-immunotherapy group (group B, n= 50), and normal control group (group C, n = 50). The levels of peripheral blood T-lymphocyte subsets (CD3+ , CD4+, CD8+ , CD4+/CD8+ ratio), natural killer cells (NK cells), immunoglobulins (IgG, IgA, IgM) and cytokines (IL-6, IL-8, TNF-a) were measured respectively before and after therapy. Group A and B were followed up for 1 year, then the times of acute outbreak and hospitalization of patients in the two groups were also compared. Results The levels of CD4 + ,CD4+/CD8+ ratio and NK cells in group A, B were significantly lower before therapy (P<0. 05~0. 01=, and the levels of IL-6, IL -8, TNF-a and IgA were significantly higher than in group C (P<0. 01=. After treatment with Mycobacterium vaccae vaccine in group A, the levels of CD4+ , CD4+/CD8+ ratio and NK cells were significantly higher (P<0. 05-0. 01= and IL-6, IL-8, TNF-a and IgA were significantly lower than before treatment (all P<0. 01=. These levels showed no significant changes in group B after treatment (P>0. 05). After 1-year follow-up, the times of acute outbreak and hospitalization on patients were statistically lower in group A than in group B (P< 0. 01 ).ConclusionsMycobacterium vaccae vaccine can improve cellular immunity function and reduce the times of acute outbreak and hospitalization in patients with stable COPD, so it has a higher clinical application value.