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 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 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 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 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: To explore the effects of whole wheat flour on blood glucose and lipid levels and antioxidant capacity of high-fat fed mice. Methods: Thirty-two male C57BL/6J mice at ages of 3 to 4 weeks were randomly divided into 4 groups with 8 mice in each group. The normal control group was fed with ordinary diet, the high-fat diet group was fed with high-fat diet, the whole wheat flour group was fed with high-fat diet and whole wheat flour, and the refined wheat flour group was fed with high-fat diet and refined wheat flour. Nine weeks later, blood was collected from the tail for measurement of fasting blood glucose (FBG), and blood was also collected from the eyeball to determine the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA). Adipose tissue was taken and weighed after death. Body weight, total food intake, Lee's index, adipose index, blood glucose, blood lipids, and antioxidant indicators were compared among the four groups of mice. Results: Compared with the normal control group, the mice in the high-fat diet group, whole wheat flour group, and refined wheat flour group exhibited increased body weight, total food intake, Lee's index, and adipose index, as well as decreased GSH-Px levels; the high-fat diet group had elevated levels of TC, TG, LDL-C and MDA (all P<0.05). When compared to the high-fat diet group, the whole wheat flour group showed lower Lee's index and adipose index, but the difference was not statistically significant (both P>0.05). Both the whole wheat flour group and the refined wheat flour group had reduced levels of TC, TG, LDL-C, and MDA, as well as increased GSH-Px levels (all P<0.05). Conclusions Whole wheat flour can effectively reduce the body weight of high-fat feeding mice, improve blood lipid levels, and enhance antioxidant capacity. However, there was no significant difference in the effects of whole wheat flour and refined wheat flour on mice during the experimental period.

In order to carry out the training objectives ofexcellent doctor education training pro-gram implemented by the Ministry of Education and the Ministry of Health, an integrated organ-system oriented curriculum system of circulatory system was applied in five year clinical medicine education. The teaching contents of circulatory system from traditional Histology and Embryology, Physiology, Pathology, Pathophysiology, and Pharmacology were integrated and optimized to compile the integrated circulatory sys-tem textbook. problem-based learning (PBL) and comprehensive morphology experimental teaching were implemented into the integrated circulatory system curriculum. Diversified examination methods including continuous assessment of PBL were used to evaluate the learning effect of students. As a result, a new cur-riculum model of circulatory system teaching was constructed. And on the base of this curriculum model, an innovation course research experiment was performed, a digital network teaching platform with 4A (anybody, anything, anywhere, anytime) was set up, and diversified teaching methods were applied. This new developing curriculum system is helpful for students to relate what they have learned in the basic sci-ences to clinical medicine, gain clinical thinking competence, lifelong skills of self-directed learning, in-novative thinking, and ability of scientific research and practice.

Aim To determine the effects of Qiliqian-gxin injected into lateral ventricle on Cardiac function, angiotensin Ⅱ( Ang Ⅱ) , angiotensin converting en-zyme(ACE), angiotensin type 1 receptor(AT1R) and the sympathetic nervous system in the hypothalamic pa-raventricular nucleus of rats with chronic heart failure. Methods Rat model of heart failure was prepared by acute myocardial infarction that was induced by ligation of the left anterior descending coronary artery. Four weeks after heart failure, Qiliqiangxin and Losartan were continuously administered via a syringe pump in-jector connected to lateral ventricle. After four weeks, echocardiogram was used to evaluate the cardiac func-tion and HE was used to observe myocardial tissue morphology, and enzyme linked immunosorbent assay was used to measure plasma norepinephrine( NE) , ser-um NT-proBNP and Ang Ⅱ in the paraventricular nu-cleus. The expression of ACE and AT1 R at mRNA and protein levels in the paraventricular nucleus was deter-mined by Real-time PCR and Western blot, and the RSNA was measured by PowerLab in anesthetized rats. Results Compared with the sham control, the cardiac function was significantly lower while the AngII, ACE, AT1 R expression in the paraventricular nucleus and RSNA were significantly increased in rats with heart failure. Compared with heart failure control, Qiliqian-gxin and Losartan decreased the RSNA and the AngII, ACE, AT1 R expression in the paraventricular nucleus. Conclusion Giving traditional Chinese medicine to the lateral ventricles can decrease the activation of the RAS system, reduce the renal sympathetic nerve activi-ty and improve cardiac function.