The cortexes were obtained from new-born rats and dissociated to single cells by triturating. The cells were cultured in neural stem cell (NSC) culture medium (DMEM supplemented with bFGF, EGF and B27) and formed primary neurospheres after 7 days. Single cells dissociated from neurosphere were cultured in 96-well plates and formed single-cell cloning neurosphere 7 days later. The primary and single-cell cloning neurospheres were both positive for the immunofluorescent staining of nestin and were identified as NSC. It was proved that NSC can be expanded in vitro and provide seed cells for neural tissue engineering.
Animals, Newborn ; Cell Separation ; Cells, Cultured ; Cerebral Cortex/*cytology ; Culture Media ; Neurons/*cytology ; Rats, Sprague-Dawley ; Stem Cells/*cytology ; Tissue Engineering

Objective To find some risk factors correlated with test anxiety of middle school students,and to find out influencing pathway for test anxiety. Methods 647 middle school students were investigated with Sarason' Test Anxiety Scale (TAS), Achievement Motivation Scale (AMS), Coping Style Scale for School Students ( CSS), Eysenck Personality Questionnaire( EPQ), Egma Minnen av Bardndosnauppforrestran(EMBU) and Family Environment Scale-Chinese Version(FES-CV). Statistics were done with version of SPSS14.0,and data were analyzed by Pearson correlation, multiple linenear stepwise regression and path analysis. Results The rates of test anxiety respectively was mild 25.97% ,moderate 45.65% ,severe 28.38%; there were no significant different between the male and female students anxiety ( 16.71 ± 6.44,17.01 ± 7.02, t = 1. 469, P = 0.334). Test anxiety positively correlated with Achievement motivation, reach motivation of competition, endurance, escape, expos, deny the fantasy,family conflicts,parental punished severely,excessive interference,objective deny,overprotective of father.( r 1-16 :0. 214,0. 135,0. 254,0. 216,0. 308,0.472,0. 492,0. 168,0. 249,0. 537,0. 282,0. 102,0. 238,0. 185,0. 233,0.301,0.273; P 1-16 = 0. 000 ～ 0. 030) , and negatively correlated with Problem-solving, rationalizing interpretation, family cohesion, informative, entertaining, emotional expression, organization, parental warmth and understanding ( r1-9: -0. 121, -0. 134, -0. 178, -0. 215, -0. 221, -0. 101, -0. 298, -0. 136, -0. 168; P 1-9 =0.000 ～0.007). Enter test anxiety regression equation is the reached motivation of competition,emotional expose,organization, psychosis, Neuroticism, parent's warm and understanding , mother's refuse and deny ( t 1-7: 2.496,2.521, -2.687, -2. 150,3.503,2.237,2.259; P1-7 =0.001 ～0.038). Conclusion Test anxiety is commonly find in middle school students. Test anxiety is affected by some paths that are personality,achievement motivation,emotional coping style,family environment and parental education methods.

ObjectiveTo find incidence rate of no-suicide self injury (NSSI) of junior school students in Dalian city,and to provide some evidences for interventions for them.MethodsRandomly drawn out 1463 junior school students were served as study objects.All objects were evaluated with Ottawa Self-Injury Inventory (OSI) and self-made investigate questionnaire,75 no-suicide self-injury were screening out.ResultsThe incidence ratio of NSSI of junior school students was 5.4％,there were no significant different between the male and female students.The highest incidence ratio was found at 13 years old.The average age for first-time self-harm was (12.24±1.344) years old.There were no significant different between the male and the female students( t=- 1.415,P =0.163 ) ; Cutting Skin was the most common way of NSSI ( 12.0％ ) ; Ideation of 80％ of the NSSI was from their Own Idea.2.7％ of the NSSI can Feel Relief through NSSI behavior.93％ of NSSI was to regulate their mood.100％ students of NSSI against NSSI behavior byReading Books or Listening Music,in which 60％ of NSSI believe the method was helpful to relax their mind.78.5％ of the NSSI resisted NSSI behavior by Watching TV or Playing Games,but they did not get enough effects.60％ NSSI considered themselves without the need to treat.41.3％ of NSSI had never been to treat.2.5％ of NSSI went to hospital for the wound.ConclusionNSSI is often be found in junior school students,and highest ratio is at 13 years old.The most common method of NSSI is Cutting Skin.More NSSI aim is to release their emotion,and self-injury behavior accordance with their inner thoughts.NSSI behavior often is secret,and reading and listening to music is cut off from the relative effective way to conduct.NSSI are seldom to initiative doctor,and education organ,parents and society in a three-dimensional one of the system is necessary.

To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor β1 (TGF-β1) gene in vitro, cultured BMSCs were transfected with the complexes of pcDNA3-TGF-β1 and Lipofectamine Reagent in vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type Ⅰ propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type Ⅰ expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-β1 gene could promote the osteogenic potential of cultured BMSCs.

To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor β1 (TGF-β1) gene in vitro, cultured BMSCs were transfected with the complexes of pcDNA3-TGF-β1 and Lipofectamine Reagent in vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type Ⅰ propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type Ⅰ expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-β1 gene could promote the osteogenic potential of cultured BMSCs.

BACKGROUND: It was thought that there was no regeneration capacityin central nerves. Recent research shows that regeneration capacity of injured neural axons and recovery of some neurological functions can be achieved by changing local surroundings after spinal cord injury (SCI).OBJECTIVE: To probe into whether the transplantation of fetal olfactory ensheathing cells (OECs) in recovering the neurological functions of patients with chronic SCI is safe, feasible, and effective.DESIGN: Auto-control observation before and after surgery.SETTING: Neurological Research and Treatment Center of Beijing Xishan Hospital; Second Department of Neurosurgery in Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences; Second Department of Neurosurgery in Naval General Hospital.PARTICIPANTS: A total of 171 patients with chronic spinal cord injury were selected from the Second Department of Neurosurgery in Beijing Chaoyang Hospital Affiliated to Capital University of Medical Sciences and the Second Department of Neurosurgery in Naval General Hospital betweenNovember 2001 and February 2003, of which there are 147 patients with complete injury and 24 ones with incomplete injury. Post-injury period ranged from 0.5 to 18 years. Process of treatment is discussed and permitted by relevant Medical Ethics Committees. Cells were obtained from voluntary donors and patients agreed to receive the treatment.METHODS: ① Fetal olfactory bulbs were cultured for 12-17 days after being digested into single cells. ② Fetal OECs were transplanted into sites rostral and caudal to the epienter. ③ Neurological functions of all patients 2-8 weeks before and after operation were evaluated according to the scoring standard of ASIA.MAIN OUTCOME MEASURES: ① Status of functional recovery in spinal cord of patients after transplantation of OECs. ② Harmful events and side effects.RESULTS: A total of 171 patients were involved in the analysis of results.①Status of functional recovery in spinal cord of patients with OECs transplantation: Partial neurological functions of 171 patients rapidly recovered,whose motor function score increased from (34.5±20.3) points before operation to (42.0±20.0) points (P ＜ 0.001) after operation, score of light touch increased from (47.2±24.0) points to (61.8±23.0) points (P ＜ 0.001) after operation,score of pain sense increased from (48.6±23.5) points to (64.0±22.8) points (P ＜ 0.001). ②Harmful events and side-effects: Early manifestations of spinal cord injury induced by infection in surgical area of one patient aggravated; two patients suffered from serious pulmonary infection,one patient from thalamic hemorrhage. Three patients mentioned above died of serious respiration and circulatory failures.CONCLUSION: OEC transplantation can rapidly promote partial neurological function of patients with chronic SCI, while the mechanism needs further observing.

Objective To find out parental locus of control and mental health affecting students test anxiety. Methods The samples were from 5 middle schools including 1000 students,and their parents. They were investigated with the general life scales, Sarason's test anxiety scale, Symptom checklist 90 (SCL-90)and Parenting Locus of Control Scale(PLOC). Results The ratio of test anxiety: the mild was 28.8%, moderate was 45.0%, severe was 26.2% ,and felt anxiety was 55.8%. Ratio of test anxiety was higher in the boy students (X2=9.284, P =0.010) ,and heavier(F:16.42±6.70; M:15.18 ±6.51, t=2.716, P=0.007). Student test anxiety was significantly positively correlated with their parental mental health (r fatherl～11=0.166～0.272, Pfather l～11= 0.000;r mother 1～11=0.182～0.242, P mother 1～11= 0.000); Student test anxiety positively correlated with the effectiveness cof parents education, father's belief on the fate, eontrol of fathers for their children, locus of control from fathers (r1～5=0.075～0.143; P1-5=0.000～0.030);felt anxiety positively correlated with self-expect ,pressure from their mothers,over take care attitude from their parents,self-pressure from their parents( r1～6=0.068～0.230; P1-6=0.000～0.050) ,and negatively correlated with respect attitude of parents for their child, Marital relations,Communication time between the students and their parents,attitude of mother for learning( r1-7=-0.074～-0.140;P1～7=0.000～0.034). Felting test anxiety was served as a dependent factor, some factors enter the regression equation,and they were somatization of father, psychotic mother, the pressure from parents, mother over expectations, self-expectations ,the child control from student's father by Logistic regression analysis ( OR1～16=0.675～3.029;P1-6=0.000～0.007). Conclusion Students test anxiety is a common problem in male and female students. Mental health and locus of control from students' parents show test anxiety has somatization of father, psychotic mother, the pressure from parents, mother' expectations,self-expectations, the control of father for the child 6 risk factors.

Objective To discuss family environment and parent's education pattern of violent online games addictive disorder in junior middle school students. Methods From August to December at 2008,6000 students were randomly drown from 8 junior middle schools in Dalian. All subjects were investigated with the self-made internet game survey form and the Chen internet addiction scale (CLAS). 149(2.5% ) internet game addicts were screen out from 6000 students, and then were divided into the violence and entertainment game addicted group (66 students in per group, and the male 55, female 11). No-game group 66 students were matched according tothe sex and the age. All subjects were evaluated with Egma Minnen av Bardndosnauppforstran ( EMBU) and Moss's Family Environment Scale of Chinese version. Results Lower mother's understand and warm were observed in violence internet online game addictive student group than other two groups( (46. 37 ± 13. 93,49. 80 ± 7.13,53. 16 ± 14. 89) , F = 4.88, P<0.01) ,and the mother understand and warm were remarkably lower in the violence online game addictive group than the non-game addicted group(P<0.01) ,and mothers rejection and the denial are obviously higher in entertainment online game addictive group than the non-game group(P = 0. 038). In family environment survey results suggested there were significant different during the 3 groups (F1-3 = 13. 99, 12.24,10.56; P<0.05)in emotion expression(4. 74 ± 1. 32,6. 06 ± 1. 97,5. 80 ± 1. 11) ,and conflict (5. 57 ± 1.85,5.28 ± 1.41,3.63 ± 1.50) .independence (3.96 ± 1. 87,4.56 ± 1.84,4.63 ± 1. 85). In the violence online game addictive group, emotion expression was lower than other two groups (P<0. 01) ,the conflict score was higher than other two groups (P<0.01). The controlling was lower than other two groups ( all P<0. 01) ; family function and total score are lower in online game addictive groups than other groups (P<0. 01, P<0. 05). Conclusion -Lower emotion expression,higher conflict,lacks of mother's love and warm,and lower controlling are found in violence online game addictive junior middle school students.

Objective To discuss the personality characteristic and personal social attributes of junior middle school students of online violent games addictive disorder (VOGAD).Methods From August to December at 2008,6000 students were randomly drown from 8 junior middle schools in Dalian ,then they were investigated with the self-made internet game survey form and the Chen internet addiction scale (CLAS).149( 2.5% ) internet game addicts were screen out from 6000 students, and were divided into the violence and entertainment game addicted group ( 66 students in per group,and the male is 55, female is 11 ).Average age was ( 14.36 ± 0.81 ) years old.No-play game group 66 students were mach according to the sex and the age.All objects were evaluated with Eysenck personality questionnaire (EPQ) and Minnesota Multiphasic Personality Inventory Abbreviation Form (MMPI-AF).Results There were significant different during the 3 groups in psychoticism (50.60 ± 5.10;50.40 ± 6.30; 46.81 ± 4.62 ), neuroticism ( 51.74 ± 5.70; 49.03 ± 6.85; 49.25 ± 6.09 ) , manifest anxiety (55.29 ±9.58;45.11 ±8.31; 43.24 ±8.32) ,dependence (58.90 ±9.09; 55.97 ±8.63; 54.05 ± 11.39) ,and social responsibility ( 37.47 ± 12.41; 43.25 ± 11.24 ;46.34 ± 14.37 ) (F1-5 = 10.32,3.83,36.17,4.11,8.23; P 1-5 ＜ 0.05 ).Except of the social responsibility (P＞ 0.05 ) ,above results all suggested higher scores in the VOGAD group than other two groups (P1 ＜ 0.05 ).Conclusion The results suggest higher scores in the psychoticism, neuroticism, dependency, manifest anxiety in VOGAD group, and lower Re in online violence game addict students.

The feasibility of using gene therapy to treat full-thickness articular cartilage defects was investigated with respect to the transfection and expression of exogenous transforming growth factor (TGF)-beta 1 genes in bone marrow-derived mesenchymal stem cells (MSCs) in vitro. The full-length rat TGF-beta 1 cDNA was transfected to MSCs mediated by lipofectamine and then selected with G418, a synthetic neomycin analog. The transient and stable expression of TGF-beta 1 by MSCs was detected by using immunohistochemical staining. The lipofectamine-mediated gene therapy efficiently transfected MSCs in vitro with the TGF-beta 1 gene causing a marked up-regulation in TGF-beta 1 expression as compared with the vector-transfected control groups, and the increased expression persisted for at least 4 weeks after selected with G418. It was suggested that bone marrow-derived MSCs were susceptible to in vitro lipofectamine mediated TGF-beta 1 gene transfer and that transgene expression persisted for at least 4 weeks. Having successfully combined the existing techniques of tissue engineering with the novel possibilities offered by modern gene transfer technology, an innovative concept, i.e. molecular tissue engineering, are put forward for the first time. As a new branch of tissue engineering, it represents both a new area and an important trend in research. Using this technique, we have a new powerful tool with which: (1) to modify the functional biology of articular tissue repair along defined pathways of growth and differentiation and (2) to affect a better repair of full-thickness articular cartilage defects that occur as a result of injury and osteoarthritis.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; cytology ; Gene Transfer Techniques ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; Stem Cells ; cytology ; metabolism ; Tissue Engineering ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics