1.SURVIVAL AND DIFFERENTIATION OF EMBRYONIC NEUROEPITHELIAL CELLS FOLLOWING TRANSPLANTATION INTO THE LATERAL VENTRICLE OF RATS
Yingmao GAO ; Jinhao SUN ; Lin YANG
Acta Anatomica Sinica 2002;0(05):-
Objective The aim of the study was to observe the survival and differentiation of embryonic neuroepithelial cells in lateral ventricle of rats after transplantation. Methods Embryonic rats(E12d) were obtained from the pregnant Sprague\|Dawley rats;neuroepithelial cells were dissociated from embryonic neural tube and treated with 0\^25% trypsin,then transplanted into the lateral ventricle of father rat.The hosts were anesthetized and transcardially perfused with PBS followed by 4% paraformaldehyde fixative by 10?d,14?d,21?d and 28?d transplantation respectively.Coronal sections of the brain were cut with in cryostat microtome.Using the techniques of histology staining,Nissl staining and immunocytochemistry, survival of neuroepithelial cells transplant and neuro\|specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression of some stem cells were examined. Results Some grafts were found survival which attached to the ventricle by day 10.Some grafts could migrate into the third ventricle and grew into a mass of cells.Some stem cells were able to differentiate into pyramidal cells.By immunocytochemistry,a mass of NSE\|positive cells and GFAP\|positive cells could be detected in the core of graft region.Around intracerebral grafts,some astrocytes showed immunoreactive for glial fibrillary acidic protein.Conclusion\ These findings demonstrate that embryonic stem cells dissociated from neural tube can survive and differentiate into neurons and astrocytes.
2.THE STUDY OF DIFFERENTIATION OF EMBRYONIC STEM CELLS AFTER TRANSPLANTATION INTO STRIATUM
Yuji GUO ; Yingmao GAO ; Jinhao SUN ; Kai LIU ; Luju BING
Acta Anatomica Sinica 1955;0(03):-
Objective To explore the survival,migration and differentiation of embryonic stem cells after transplantation into striatum and provide advantageous data for feasibility and safety of therapeutic transplantation. Methods We transplanted embryonic stem cells(undifferentiated ESCs and ESCs that had already developed into the stage of neural progenitor cells respectively) into striatum of the rat.Then differentiated cells were determined by morphological observation,Nissl's staining,TH and BrdU immunocytochemistry. Results We found that after transplanted 4 weeks,partially differentiated ESCs could survive and migrate into the surrounding host tissue.Some of them differentiated into TH-positive cells which had Nissl's bodies in cytoplasm.Whereas undifferentiated ESCs couldn't differentiate into TH-positive cells effectively and have the tendency to form tumor.Conclusion When conducting transplantation experiments of ESCs,it's better for ESCs to be induced into the stage of neural progenitor cells first and then transplanted.;
3.AN EXPERIMENTAL STUDY ON THE THERAPY OF PARKINSON DISEASE BY NEUROEPITHELIAL CELL TRANSPLANTATION INTO RAT BRAIN
Jinhao SUN ; Yingmao GAO ; Lin YANG ; Zhenhua LI
Acta Anatomica Sinica 1954;0(02):-
Objective Embryonic neuroepithelial cells (NEC) were transplanted into the brains of Parkinson disease rats, the survival and differentiation of NEC were investigated. Methods Embryonic rats (E11) were obtained from the pregnant Wistar rats. Neuroepithelial cells were dissociated from embryonic neural tube and treated with 0^25% trypsin, then transplanted them into the striatum of Parkinson disease (PD) rats stereotaxicly. Coronal sections of the brain were made in cryostat. The survival of transplant neuroepithelial cells and tyrosine hydroxylase (TH) expression of some NEP were examined by immunocytochemical technique, also the rotation behavior was tested following different time of transplantation. Results Some grafts were found survived well and some TH-positive cells could be detected in the core of graft region. NEP transplantation could also partly improve the symptoms of PD rats.Conclusion The results demonstrate that embryonic stem cells dissociated from neural tube can survive and differentiate into dopaminergic neurons. NECs contribute to the behavioral recovery.
4.PREGNANCY INCREASES CELL PROLIFERATION AND NEUROGENESIS IN THE MOUSE DENTATE GYRUS
Zhengang YANG ; Lin YANG ; Yingmao GAO ; Jinhao SUN ; Zhenhua LI ;
Acta Anatomica Sinica 1953;0(01):-
Objective To observe cell proliferation and neurogenesis in the pregnant mouse dentate gyrus (DG) and the subventricular zone(SVZ). Methods Injection of the thymidine analog 5 bromo 2 deoxyuridine (BrdU)to determine the extent of cell proliferation combined with single or double immunohistochemistry staining with antibodies BrdU,TuJ1(class Ⅲ ? tubulin,neuron specific early differentiation marker)and GFAP. Results The number of BrdU positive cells in the pregnant mouse dentate gyrus was significantly more than that of unpregnant mouse dendate gyrus but not in the subventricular zone.In dentate gyrus,approximately 80% of these cells were neuronal characteristics (TuJ1 immunoreactive)and 3%~5% of these cells were astrocytic characteristics(GFAP immunoreactive).Conclusion\ These findings suggest that pregnancy significantly increases cell proliferation and neurogenesis in the adult mouse dentate gyrus and present the possibility that these new cells exert an important influence on hippocampal function
5.ISOLATION AND INDUCED DIFFERENTIATION OF NEUROEPITHELIAL STEM CELLS FROM SPINAL CORD NEURAL TUBE
Zhengang YANG ; Lin YANG ; Yingmao GAO ; Jinhao SUN ; Zhenhua LI ;
Acta Anatomica Sinica 1953;0(01):-
Objective To isolate neuroepithelial stem cells from the spinal cord neural tube of the embryonic rat and induce them to differentiate into dopaminergic neurons. Methods Serum free cells suspension culture and single cell cloning technique were used to isolate neuroepithelial stem cells. 5 bromo 2 deoxyuridine(BrdU) to label new cells combined with single or double immunocytochemistry staining to detect nestin antigen before differentiation and neural cell specific antigens after differentiation, such as neurofilament (NFM 160?kD), glial fibrillary acidic protein(GFAP), galactocerebroside(GalC) and tyrosine hydroxylase(TH). Striatal extracts were used to induce neuroepithelial stem cells to differentiate into dopaminergic neurons. Results The cells isolated from the spinal cord neural tube of the embryonic rat expressed nestin antigen. They had the potential to serially passage and differentiate into neurons, astrocytes and oligodendrocytes. Striatal extracts could induce 12% of them to differentiate into dopaminergic neurons compared with 3% in controls.Conclusion The cells, which express nestin antigen, isolated from neural tube are multipotent and have the ability to self renew, therefore, they are neural stem cells. These stem cells can be induced to differentiate into specific neurons in vitro. Which can provide materials for neural transplantation.
6.The expression of miR-133b in the brain of methamphetamine dependent rats and its regulation on neuronal toxic injury
Tao LI ; Haili LIU ; Hui WANG ; Mingfei WANG ; Hongjie WANG ; Guiyang LIU ; Bo ZHANG ; Jinhao SUN
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(8):689-694
Objective To investigate the expression of miR-133b in the brain of methamphetamine(MA) dependent rats and its regulatory effects on neuronal toxic injury.MethodsThrough continuous intraperitoneal injection to rats with MA(10 mg/kg),the conditioned place preference(CPP) rats model was established,and the expression of miR-133b in the cerebral cortex of model rats was detected by real-time fluorescence quantitative PCR(RT-PCR).PC12 cells were cultured in vitro and treated with MA(800 μmol/L),and then miR-133b expression in cultured neurons was detected.miR-133b mimics and inhibitor were transfected to PC12 cells respectively to observe the effect of miR-133b on the mitochondrial membrane potential(MMP) of cultured neurons.ResultsAfter continuous intraperitoneal injection with MA for 14 days,the residence time of rats in the box with medicine((620.20±44.80)s) was significantly longer compared with the control group((341.80±25.12)s,P<0.01),which showed that MA dependent rats model was successfully established.The RT-PCR detection results showed that the expression of miR-133b in the cerebral cortex of model rats(0.36±0.05) significantly decreased compared with the control group(0.99±0.08,P<0.01).In the in vitro model,most of the neuronal cell bodies became round and the neuorites were withdrawn after MA treatment.Compared with the control group(1.00±0.02),the RT-PCR detection results showed that the expression of miR-133b in MA group(0.74±0.05) decreased(P<0.05).The JC-1 detection results showed that the MMP of the MA group(109.85±7.03) decreased significantly contrast to the control group(36.49±3.89,P<0.01),the MMP of the miR-133b mimics group(58.97±6.56) increased significantly contrast to the mimics control group(135.46±15.04,P<0.01) and the MMP of the miR-133b inhibitor group(162.84±14.15) decreased contrast to the inhibitor control group(139.81±12.26,P<0.05).ConclusionsThe expression of miR-133b in the cerebral cortex of MA dependent rats and in vitro neuron model treated with MA are significantly downregulated.By regulating the expression of miR-133b,the MMP damage of cultured neurons treated with MA is changed,indicating that miR-133b is not only involved in the nerve injury induced by MA,but also possiblely as a molecular target for intervention.
7.Expression of miR-134 in Methamphetamine-induced Neuronal Injury and Its Effect on Evoked Action Potential
Tao LI ; Hongjie WANG ; Guiyang LIU ; Bo ZHANG ; Wenbo ZHAO ; Tianwei LIU ; Kaiwen ZHU ; Jinhao SUN
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(1):36-41
[Objective]To investigate the expression change of miR-134 in methamphetamine(MA)-induced neuronal injury in PC12 cells and its effect on neuronal excitability and understand the pathogenesis of methamphetamine-induced neuronal injury.[Methods]PC12 cells in the logarithmic phase were divided into control group and MA group. The MA group was treated with 800μmol/L MA to establish the model of neuronal injury. The cellular injury was observed under microscope. The neuronal apoptosis was detected by Hoechst3342/PI double staining,and miR-134 expression was measured by using real-time quantitative PCR (Real time-PCR). Furthermore,we constructed miR-134 interference vector and observed its effect on evoked action potential.[Results]The cultured PC12 cells were damaged under the 800 μmol/LMA treatment ,and neurites became shorter ,the apoptotic cells were evidence. Real time-PCR showed that miR-134 expression was increased after MA treatment. Electrophysiological data showed that the evoked action potential increased after miR-134 interference.[Conclusions]High concentration of MA can induce neuronal damage and apoptosis and also increase miR-134 expression. While silence miR-134 expression can increase neuronal excitability.Our study provides an experimental basis for elucidating the possible mechanism of MA-induced neuronal injury and the role of miR-134 in neurotoxicity and neuronal excitability.
8.The clinical comparative study on the therapeutic effects of NICU patients implemented by NICU professional doctors and non-NICU professional doctors
Linyue GUO ; Peng WANG ; Chuang GAO ; Wanqiang SU ; Jinhao HUANG ; Yu QIAN ; Jiaqi WANG ; Zhitao GONG ; Yiming SONG ; Jian SUN ; Rongcai JIANG
Tianjin Medical Journal 2017;45(8):833-837
Objective To explore the implementation styles on the therapeutic effects on the neurosurgical intensive care unit (NICU) patients. Methods Patients were enrolled during February 3, 2015 to February 3, 2016. The key point time was August 3, 2015 when the treatment in our NICU was fully implemented by NICU professional doctors. Based on this time point, all the enrolled patients were divided into non-NICU professional doctor implementing (NNPDI) group and NICU professional doctor implementing (NPDI) group. Thus non-NICU professional doctors and professional doctors were the leaders of diagnosis and treatment in tow groups. The length of hospital stay, complications, prognosis and other therapeutic outcomes were compared between two groups. Results The length of hospital stay was longer in NPDI group than that in NNPDI group (P<0.05). The incidence of water-electrolyte imbalance was lower in NPDI group than that in NNPDI group (P<0.05). There were no significant differences in the incidence of the ventilator-associated pneumonia (VAP), the hepatic and renal insufficiency, the intracranial infections and stress ulcers between the two groups (P>0.05). The proportion of referral to other wards and fatality rate were both lower in NPDI group than those in NNPDI group (P<0.05). And the discharge rate from NICU was higher in NPDI group than that of NNPDI group (P<0.05). There was no significant difference in the rate of patients left hospital without treatment between the two groups (P>0.05). Conclusion The NICU professional doctor implementing may be contribute to, at least in part, the improving of prognosis of NICU patients without obvious advantages in most complications. The level of professional management remains to be improved.
9.The reproducibility of anterior, middle and posterior diaphragm motion assessments with ultrasound
Yelin YAO ; Yinghua SUN ; Jinhao TAO ; Kang CHEN ; Sujuan WANG
Chinese Journal of Physical Medicine and Rehabilitation 2020;42(7):601-605
Objective:To explore the reproducibility of ultrasound measurements of children′s anterior, middle and posterior diaphragm motions.Methods:Thirty children admitted to a pediatric intensive care unit were positioned supine and a 5MHz ultrasound probe was placed over the intersection of their right midclavicular line with the costal margin. M-mode ultrasound was used to record the excursion and contraction velocity of the anterior, middle and posterior diaphragm during respiration. The observations were duplicated so the repeatability of the measurements could be evaluated using intra-group correlation coefficients calculated for the diaphragm excursions and the contraction velocities. Analysis of variance was used to explore the differences in excursion and contraction velocity among different parts of the diaphragm.Results:The intra-group correlation coefficients calculated for the anterior, middle and posterior diaphragm were 0.89, 0.95 and 0.90 respectively. The corresponding values for the contraction velocities were 0.90, 0.94 and 0.95 respectively. Both variables measured by ultrasound showed high repeatability. The average anterior, middle and posterior diaphragm excursion values (in mm) were 8.1±3.1, 7.4±3.0 and 5.5±2.3, and the corresponding average contraction velocities (in mm/s) were 12.5±4.8, 11.5±6.3 and 8.9±4.0.Conclusions:Measurements of children′s diaphragm motions using ultrasound show high repeatability. The excursions and contraction velocities of the anterior, middle and posterior diaphragm differ in children. The motion of one part of the diaphragm cannot represent the functioning of the entire diaphragm.
10.Clinical analysis of the changing trend of serum thyrotropin in patients after hemithyroidectomy
Zhihong WANG ; Hao ZHANG ; Ping ZHANG ; Wei SUN ; Jinhao LIU ; Yuan QIN ; Liang HE ; Wenwu DONG ; Dalin ZHANG
Chinese Journal of Endocrinology and Metabolism 2017;33(11):955-958
Objective To evaluate the changing trend of serum thyrotropin (TSH) levels for hemithyroidectomy patients,and to discuss the necessity and strategy of TSH suppression for low-risk differentiated thyroid carcinoma(DTC). Methods One hundred and twenty-seven patients with benign thyroid nodules undergoing hemithyroidectomy between January 2013 and June 2014 were retrospectively studied. Serum thyroid hormones levels FT3,FT4,TSH,thyroid peroxidase antibody(TPOAb),and thyroglobulin antibody(TGAb)were detected at 1 month after surgery for all patients and at 3 month for 54 patients. Results (1)Mean TSH level at 1 month after surgery was significantly higher than preoperative TSH level(2.45 mIU/L vs 2.20 mIU/L,n=127,P<0.01). The mean TSH level at 3 month after operation was significantly higher than preoperative ones(2.46 mIU/L vs 2.35 mIU/L,n=54, P<0.05). (2)TSH<2. 0 mIU/L was found in 52 patients(40. 9%) and TSH>4. 94 mIU/L in 18 patients (14.17%) at 1 month after operation. TSH<2.0 mIU/L was found in 28 patients(51.85%)and TSH>4.94 mIU/L in 8 patients(14.81%) at 3 month after operation. (3)A preoperative TSH≥2.0 mIU/L and the coexistence of Hashimoto's thyroiditis were found to be independent risk factors for the TSH levels higher than 2.0 mIU/L. Among the patients with TSH≥2. 0 mIU/L at 1 month, 13 exhibited spontaneous recovery at 3 month, coexistence of Hashimoto's thyroiditis was related to this phenomenon. Among the patients with TSH<2.0 mIU/L at 1 month,TSH levels were elevated over 2. 0 mIU/L in 7 patients by 3 month comparing to that by 1 month. Coexistence of Hashimoto's thyroiditis was independent risk factor for the TSH elevation. Conclusion TSH suppression may still be performed to patients with low risk DTC after operation especially to whom the preoperative TSH≥2.0 mIU/L and the coexistence of Hashimoto's thyroiditis. Suppression therapy should be carefully considered with close follow-up.