Objective To study the correlation between bone marrow edema (BME), the amount of joint fluid and clinical symptoms in order to strengthen further understandings about clinical significance of BME in osteonecrosis of the femoral head (ONFH). Methods Fifty-seven patients (91 hips) with ONFH proved by clinical follow-up or pathology were examined by conventional radiography, 1.5 T MRI, and radionuclide imaging. BME, necrotic area, and joint fluid were analyzed respectively in ONFH with pre- or post-collapse of the femoral head and different MR signal intensities within necrotic area.Results ①The characteristic “line-like sign” appeared on MRI in 88 of 91 affected hips, and BME was seen in the distal zone away from line(s) in 61 hips, extending to the femoral neck and intertrochanteric region. ②The ratio of the occurance of BME in the collapse was greater than that in noncollapse, and in mixed signals within necrotic area without collapse greater than pure fat-like signal (P0.05), and both were greater than noncollapse without BME (P

Objective To explore the clinical characteristics of poststroke epilepsy and to discuss the association with the type of stroke,the location of focus.Methods Clinical information of 64 patients with poststroke epilepsy taken from 932 patients with stroke were analyzed retrospectively.Results In the patients with first stroke,the incidence of poststroke epilepsy was 6.9％.Among them,47 cases belong to early seizure (73.4％) and 17 cases belong to late seizure (26.6％).The incidence of poststroke epilepsy was difference between ischemic stroke and hemorrhagic stroke.There were more cases of stroke in cortical focus than subcortial one.Onset seizures type was more cases of partal seizure than generalized tonic-seizure (GTS).The treatment on early seizure could be short-range treatment and the treatment on late seizure should be long-term standardized treatment and regular.Conclusions Occurrence of poststroke epilepsy were involved in the type of stroke,the location of focus (cortex / subcortex).It is helpful for guiding the clinical therapy of patients with stroke and improving the quality of their life to analyse the clinical characteristics of poststroke epilepsy.

Objective To study normal imaging features of choroid fissure and to improve the ability of diagnosis of choroid fissureneuroepithelial cyst.Methods The MR manifestations of choroid fissure were studied by comparing general brain specimen with MR images of normal brains.The analysis of CT and MRI findings of choroid fissure neuroepithelial cysts in 14 cases were also conducted.Results The whole choroid fissure was clearly displayed as fissures full of cerebrospinal fluid on MRI.Cysts were shown round or ellapse foci with sharpmargins and homogeneous low density on CT or cerebrospinal fluid-like signal intensity on MRI.There was no enhanced contrast or nodramatic changes in follow-up studies.Conclusion The recognization of MR anatomy of choroid fissure and imaging manifestations ofchoroid fissure neuroepithelial cysts can improve the ability of diagnosis and differential diagnosis of cysts.

BACKGROUND:There are some disadvantages in harvesting and transferring cells in the traditional tissue engineering technique,and it is difficult to form dense tissues,which significantly limits the development of tissue engineering.OBJECTIVE:To explore the culture and fabrication of dog bone marrow mesenchymal stem cell(BMSC)sheet in vitro.METHODS:Bone marrow was extracted from dogs following anesthesia.BMSCs were isolated with the method of density gradient centrifugation in vitro.BMSCs at passage 4 at a density of 1×10~9/L were incubated in the temperature-responsive culture dishes with a diameter of 3.5 cm,and cultured in an incubator at 37 ℃,5% CO_2 and saturated humidity.The temperature of the incubator was changed from to 37 V to 20 ℃ to prepare BMSCs cell sheet for 20 minutes.Cell morphological changes and cell sheet formation were observed under an inverted microscope.RESULTS AND CONCLUSION:Dog BMSCs following 24 hours of primary culture presented ellipse or polygonal shape.Most cells adhered at hour 72,and cell colonies were visible at day 7.Cells showed long spindle and completely confluence at day 12,with unclear boundary.BMSCs in the temperature-responsive culture dishes presented short spindle shape,and gradually separated from the dish bottom,forming entire cell sheet containing extracellular matrix at 20 V.These verified that dog BMSCs can be effectively obtained with method of density gradient centrifugation.Complete cell sheet layer can be fabricated with temperature-responsive culture dishes.

BACKGROUND: How to reconstruct tissue-engineered bone with structure similar to natural bone iS a problem in the development of tissue engineering. Cell sheet engineering technology enables novel approaches to construction of tissue-engineered bone. OBJECTIVE: To observe the biocompatibility of call sheets to decalcified bone matrix (DBM) and their growth on DBM. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory, Affiliated Hospital, Qingdao University Medical College between June and September 2009.MATERIALS: Dog bone marrow stromal cell sheets were prepared using temperatura-responsive medium. Dog DBM was prepared by defatting, decalcification, and noncotlagen protein removal procedures. METHODS: DBM surface was covered by call sheets prepared by temperature-responsive technology and cultured with DMEM containing 10% fetal bovine serum and osteoinductive agent.MAIN OUTCOME MEASURES: Under scanning electron microscope, DBM structure, as well as the attachment and growth of cell sheets on DBM surface, was observed. Porosity and aperture size of DBM were calculated. RESULTS: DBM exhibited a three-dimensional latticed structure, with a porosity of approximately 75%. The mean aperture size was (250.11±98.89) μm, exhibiting a normal distribution. Cell sheets well attached to and grew on DBM surface, and rapidly proliferated.CONCLUSION: Cell sheets show good biocompatibility to DBM. DBM/cell sheets complex can be applied in tissue-engineered bones, which promotes the construction of tissue-engineered bone with structure similar to natural bone.

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

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.

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.

The pathogenesy of diabetic peripheral neuropathy(DPN) is approached according to collaterals diseases theory in this study,indicated that defi ciency of both vital energy and yin is the chief pathologic foundation and obstruction of collaterals by blood stasis and phlegm is the critical element in DPN.Furthermore,highlight of differentiation of symptoms and signs,therapeutic principle and diagnosis and treatment based on differentiation are illuminated.This study has supplied a new idea for precaution and treatment of DPN.

Objectiye To measure longitudinal (T1) and transverse (T2 ) relaxation time of metabolites in m. soleus (SOL) and m. tibialis anterior TA of healthy volunteers at 3.0 T through 1H-MRS and optimize measurement protocols. Methods Altogether 24 healthy volunteers were recruited in the study. All subjects signed a letter of informed consent. After divided into 2 groups randomly by the table of random number, 1H-MRS measurements with stimulated echo acquisition mode (STEAM) sequence were undertaken in SOL and TA separately. Progressive saturation method was used for T1 measurement. Spectra with 8 different TRs (770,900,1000, 1100,1200,1500,2000 and 3000ms ) were acquired with TE=20 ms.T2 time was measured by changing TE. Altogether 8 TEs (20,30,45,60,90,135,200 and 270 ms) were used with TR = 3000 ms. Metabolites' concentration was calculated through T1 and T2 correction using water as internal reference. The t test was used for statisties. Results Altogether 22 groups of data were gained ( 12 for SOL, 10 for TA ) . T1 value of water, Creatine-CH3 ( Cr3 ), Trimethyl amonium ( TMA ),extramyocellular lipid (EMCL) and intramyocellular lipid (IMCL) in SOL were ( 1384. 0 ± 36. 9 ),( 1064. 0 ± 167.0), (964. 2 ± 144. 0 ), ( 373.0 ± 46. 8 ), ( 374. 7 ± 20. 6) ms respectively and T2 value were (26.5 ±1.2), (100.2±19.3), (149. 1 ±32.7), (81.4±5.2), (84.7±4.2) ms. InTA T1 value of water, Cr3, TMA, EMCL, and IMCL were ( 1307. 0 ± 24.4), (945.7 ± 132. 0), (968.3 ± 127. 0),(372. 7 ± 39. 2), (412. 8 ±80. 2) ms respectively and T2 value were (27. 1 ± 0. 9), (135.3 ± 18. 2 ),(62.1 ± 6. 0), ( 84. 3 ± 4. 0 ), ( 90. 7 ± 3.2 ) ms. After corrected by the calculated relaxation times, the concentrations of Cr3 in SOL and TA were (33. 1 ± 3.7) and (31.7 ± 3. 1 ) mmol/kg respectively, TMA (35.2±3.2) and (32.9 ±5.2) mmol/kg, EMCL (12.2 ±5.0) and (8.9 ±4.9) mmol/kg, IMCL (9. 0 ± 2. 4) and (3.0 ± 0. 8 ) mmoL/kg. IMCL in TA was much lower than SOL with statistical significant ( t = 8. 044, P < 0. 01 ), the difference between other metabolites were not statistically significant( t = 0. 926,1. 264, 1. 542, P > 0. 05 ) . Conclusions Accurate relaxation time was measured at 3.0 T of the metabolites in skeletal muscles of healthy adult human. After corrected by the relaxation times, the absolute concentrations calculated were consistent with the reported results. Quantitative knowledge of muscle NMR relaxation time was a prerequisite for absolute quantification of metabolites using the 1H-MRS and also was useful for optimizing measurement protocols.