Objective Hair follicle stem cells (HFSCs) can be cultured in vti ro to differentiate into chondrocytes, osteoblasts and adipocytes .The aim of the article was to investigate the culture method of HFSCs and their adipogenic and osteogenic differentiation potentials. Methods Microdissection and two-step enzyme digestion were applied to obtain HFSCs , and HFSCs are purified by differ-ential adhesion method.The cells were cultured in keratinocyte serum-free medium obtaining 10% fetal bovine serum.Flow cytometry was used to detect CD34、β1-integrin and CK15.The third-generation HFSCs were cultured by osteogenic inductor and adipogenic induc-tor respectively . Results Cultured HFSCs were homogeneous in shape , bearing the property of great refraction and typical cobble-stone-morph.By flow cytometry, the expression rates of CD34,β1-integrin and CK15 were 59.6%, 97.0%and 61.1%.Stem cells were in good growing condition from the third-generation cell growth curve.21 days after being induced by osteogenic inductor, calcified nodules appeared after alizarin red staining, while 14 days after being induced by adipogenic inductor, intracellular lipid droplets could be observed after oil red O staining. Conclusion Cultured HFSCs are chaterized by strong growth capacity and multiple differentiation potential .

BACKGROUND:The rise of tissue engineering has opened up new ways for tissue repair and reconstruction of the urinary tract, and the bladder acellular matrix is a better alternative material for urinary tissue engineering.
OBJECTIVE:To construct the compound of hair fol icle stem cells with heterologous bladder acellular scaffold, and to observe the growth of hair fol icle stem cells on the scaffold.
METHODS:Bladder acellular matrix from New Zealand rabbits were prepared and detected using scanning electron microscopy and Masson staining. Passage 3 hair fol icle stem cells were statical y inoculated into the surface of bladder acellular matrix using secondary sedimentation method. Under inverted microscope, cellgrowth was observed, and cellgrowth curves were drawn. cellgrowth on the scaffold surface was observed through histological detection and scanning electron microscope observation.
RESULTS AND CONCLUSION:Prepared bladder acellular matrix was a white translucent film with fiber mesh structure, and no residual cells were seen. Masson staining results indicated that the bladder acellular matrix had col agen structure, and no obvious residual cells. After culture for 48 hours, hair fol icle stem cells grew wel around the bladder acellular matrix under inverted microscope;1 week later, hair fol icle stem cells extended and adhered to the scaffold surface. These findings indicate that hair fol icle stem cells have a good biocompatibility with the bladder acellular matrix through in vitro culture.

BACKGROUND:The hair fol icle stem cells are usual cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum. Recently, keratinocyte serum-free medium has been used to culture hair fol icle stem cells.
OBJECTIVE:To observe the effects of three different culture media on the proliferation and purity of hair fol icle stem cells.
METHODS:After sterling, the skin of the rat’s beard was cut off, and the vibrissa fol icles were dissected under a stereocroscope. The fol icles were digested by Dispase Ⅱ firstly and by the mixture of trypsin and EDTA secondly. cellsuspension was divided into three pieces based on the average number of cells and respectively cultured by keratinocyte serum-free medium, Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum and keratinocyte serum-free medium containing 10%fetal bovine serum. Hair fol icle stem cells were selected by rapid adherence on col agen type Ⅳ, and passaged.
RESULTS AND CONCLUSION:Cultured hair fol icle stem cells spread to the third generation, and trypan blue assay showed there was no difference in cellsurvival rate among the three groups (P>0.05). cellCounting Kit 8 assay showed that the cells grew slowly in the three groups in the first 2 days. At day 4, the hair fol icle stem cells grew into the growth logarithmic phase, and the proliferative activity was ranked as fol ows:keratinocyte serum-free medium+10%fetal bovine serum>Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10%fetal bovine serum>keratinocyte serum-free medium with a significant difference between the three groups (P<0.05). Flow cytometric examination showed the expression of CD34 in the keratinocyte serum-free medium+10%fetal bovine serum group was lower than that in the keratinocyte serum free medium group (P<0.05). The expressions of CD34, CK15 andβ1-integrin (CD29) in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10%fetal bovine serum were lower than those in the other two groups (P<0.05). These findings indicate that we can obtain higher purity hair fol icle stem cells in the keratinocyte serum-free medium+10%fetal bovine serum than in the Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12+10%fetal bovine serum.

[Objective]To investigate results of surgical treatment of intradural tumors of the cervical spinal cord.[Method]Twenty-one cases with cervical intradural tumor were treated surgically under posterior approach from 1999 to 2005,all patients were performed cervical laminae resection before tumor resection,and some received internal fixation.All patients were followed up for 8 months to 38 months respectively.All the clinical materials were analyzed retrospectively.[Result]All patients survived the operation,symptoms disappeared 13,relieved 7 and deteriorated 1.Total resection in 15 cases,subtotal 4 cases and partial 2 cases.[Conclusion]To succeed the operation,it is very important to make clear the location and size of the tumor and the relationship between the tumor and spinal cord;in order to gain total resection of tumor,both careful protection of spinal cord and vertebral artery and intraspinal cannal veins are essence.

Objective To compare the effect of different dosage of verapamil and a cocktail therapy(verapamil 200 ?g plus nitroglycerin 200 ?g) in the prevention of radial artery spasm(RAS) during transradial PCI.Methods It is a prospective,randomized and double-blind clinical trial.Patients who received transradial coronary intervention were divided into three groups: group A(verapamil 200 ?g),group B(verapamil 1mg) and group C(verapamil 200 ?g plus nitroglycerin 200 ?g).Different drug protocols were given randomly to the patients after sheath insertion.The diagnostic criteria is clinical definition of RAS documented by angiography.The incidence of RAS and adverse effects in each group was compared.Results A total of 621 patients were enrolled,and there were 205 in group A,207 in group B and 210 in group C.The baseline characteristics were of no difference among the three groups.Univariate analysis showed that the incidence of RAS in group A is higher than that in group B(17.1% vs.10.2%,P=0.045) and in group C(17.1% vs.9.5%,P=0.029),but there was no statistical difference in the RAS incidence between group B and C(10.2% vs.9.5%,P=0.870).Binary logistic regression analysis showed that the relative risk of RAS in group B decreased by 32.1% compared with group A(P=0.038),and the relative risk in group C decreased by 43.8% compared with group A(P=0.017).The incidences of adverse effect were similar in groups A and C,but was higher in group B when compared with group A(9.7% vs.2.4%,P=0.003) and C(9.7% vs.3.8%,P=0.019),respectively.Conclusion Verapamil 200 ?g plus nitroglycerin 200 ?g is recommended to prevent RAS during transradial coronary intervention in Chinese.

Objective The animal model of rheumatoid arthritis was established in Lewis rats by a single intradermal injection of pristane. Methods The degree of pathogenicity and the pathological characteristics were assayed by histiopathological techniques and joint roentgenography. Results The incidence of arthritis in rats immunized by pristane was 62.5%. Peripheral joints were mainly involved. A chronic relapsing and progressive arthritis was developed.As showed by histopathology and roentgenography, typical arthritis pathology such as synovial proliferation, articular cartilage and bone erosien were found. Conclusion Pristane induced arthritis mirrors human rheumatoid arthritis and is a very good animal model for rheumatoid arthritis.

Objective To discuss the curative effect of arthroscopiclly assisted radiofrequency tenotomy of the sternocleidomastoid muscle for the treatment of congenital muscular torticollis in teenagers. Methods A total of 12 patients (right, 4 patients; left, 8 patients) were enrolled in the study. A 3-mm skin incision was made at the site 5 cm below the sternoclavicular joint. A dissector was inserted through the incision to perform blunt dissection subcutaneously until the attachment points of the sternocleidomastoid muscle. After a subcutaneous operative space 3 cm ? 3 cm in size was created, a 30? wide-angle arthroscope was placed. Another incision was located at the site 5 cm below the mid-clavicle for placing the bipolar radiofrequency probes (Arthrocare 2000). Under the assistance of arthroscopy, the sternocleidomastoid muscle was severed from the attachment at the both sides. Results The operation time was 15~30 min (mean, 20 min). No intraoperative hemorrhage or vascular and nervous injuries were observed. Follow-up for 5~10 months (mean, 7 months) in the 12 patients found normal appearance and no recurrence or diplopia. Conclusions Arthroscopiclly assisted radiofrequncy therapy for congenital muscular torticollis is characterized by simplicity of manipulation, micro-invasion, and satisfactory curative effect.

Objective To study the compatibility and feasibility of construct tissue engineer bladder through biocompatibility of hair follicle stem cells (HFSCs) and heterogeneous bladder acellular matrix (BAM) in vitro and vivo.Methods The third-generation of rat HFSCs were cultured with the two-step enzymatic and different adhesion time method.The cells were identified by flow cytometry through detection expression of β1 integrin FITC-Conjugated.The New Zealand rabbit BAM were decellularized by the method of microdissection and chemical washing,and then examined by Masson staing and electron microscope to confirm no cell elements remained.The HFSCs of the rats were implanted in BAM and cultured for about 24 hours.Then the cells growth conditions on the material surface were examined by histology and scanning electron microscopy.The cells-scaffold composites were implanted in rats subcutaneously,samples and histological examination were harvested at 1,2 and 4 weeks after implantation.Results The BAM was white and translucent membranous.There were fiber network structures without cell elements remained under the examination of electron microscope.And the BAM prompted for collagen tissue composition under Masson staining,without significant residual cells.The growth condition of HFSCs beside the BAM was well that observed by inverted microscope at 48 h of co-culture.After 1 week the HFSCs extended and adhered to the matrix surface observed under the scanning electron microscope.No significant inflammatory response in rat subcutaneous implantation experiments,the single-layer cell structure in histological examination could be seen after 1 week,and multi-layer cell structure in histological examination could be seen after 4 weeks of implantation.Conclusion The biocompatibility of HFSCs and heterogeneous BAM is good,which provides a good experiment support for HFSCs to repair the bladder defects disease.

BACKGROUND:Liver fibrosis is a kind of chronic and active disease that is caused by various causes and characterized by the excessive accumulation of extracelular matrix. At present, use of Chinese herbs for the treatment of hepatic fibrosis has obvious advantages. Salvia miltiorrhiza has been shown to be highly effective in the treatment of hepatic fibrosis. However, the underlying mechanism needs further investigation. OBJECTIVE:To investigate the effects of TanshinoneⅡA on the expression levels of transforming growth factor-β/Smads signaling pathway related factors transforming growth factor-β, bone morphogenetic protein 7, Smad6 and Smad7 in the liver tissue of rats with hepatic fibrosis. METHODS:SD rats were randomly divided into three groups (n = 10): normal control, model and TanshinoneⅡA-treated groups. Rats in the model and TanshinoneⅡA-treated groups were subtaneously injected with olive oil-diluted 10% CCl4 ( 5 mL/kg) twice a week, 8 weeks in total, to build rat models of hepatic fibrosis. Four weeks after hepatic fibgrosis induction, rats in the TanshinoneⅡA-treated group received subtaneous injection of TanshinoneⅡA til eight weeks. Rats in the normal control group were subcutaneously injected with olive oil. RESULTS AND CONCLUSION: Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) detection showed that in the model group, the expression of transforming growth factor-β in the rat liver tissue was significantly increased (P < 0.01) and the expression of bone morphogenetic protein-7, Smad6 and Smad7 was significantly decreased (P < 0.01) compared with the normal control group. TanshinoneⅡA could obviously reverse the expression of those factors above-mentioned (P < 0.01). The results suggest that TanshinoneⅡA can be used for treatment of hepatic fibrosis by decreasing the expression of transforming growth factor-β and increasing the expression of bone morphogenetic protein-7, Smad6 and Smad7.

Objective To verify the validity of the sucrose density gradient ultracentrifugation method for lipid rafts from cerebral cortex. Methods Extract lipid rafts from cerebral cortex in mouse were extracted by the sucrose density gradi-ent ultracentrifugation method. The properties of lipid rafts were detected by Western blotting method, double enzyme and light scattering methods. HPLC MS/MS proteomics and bioinformatics were used to locate proteins of lipid rafts in cells. Re-sults Lipid rafts from cerebral cortex were provided with the model properties of lipid rafts such as high light scattering and cholesterol and high expression of Flotillin-1. HPLC MS/MS proteomics identified total 647 proteins. Most of these pro-teins were from plasma membrane, endoplasmic reticulum, cytoskeleton and cytosol, however, there were 21% proteins among total 647 proteins were from nucleus, mitochondria and ribosomes. Conclusion The sucrose density gradient ultra-centrifugation method is a effective method to extract lipid rafts from cerebral cortex, however, the properties of mixture should be considered.