Objective To investigate the inhibitory effect of a disintegrin and metalloprotease 12 silenced by shR?NA on self-renewal capacity of CD133 positive giloma cells. Methods The shRNA recombinant lentivirus aimed at si?lencing ADAM12 was prepared. Human glioma cells U87 were employed in this study and assigned into three groups:shRNA-ADAM12, shRNA-NCandshRNA-C. ADAM12 expression was detected at mRNA and protein level using Re?al-time quantitative-PCR and western bloting, respectively. U87 cells were cultured with stem cell culture medium, to obtain cell sphere formation in which CD133 positive glioma cells were enriched. Immunofluorescence was employed to detect the expression of ADAM12 and CD133 in cell spheres and U87 cells; Self-renewal was tested by using tumor sphere formation assay. Molecular markers for differentiated or undifferentiated cells (CD133,GFAP and Tuj1) were de?tected at protein using western blotting. Western blotting was employed to test protein expression of HES1. Results AD?AM12 shRNA significantly down-regulated the mRNA and protein expression levels of ADAM12. Compared with shRNA–C group, the relative expression levels of mRNA in shRNA-ADAM12 group and shRNA-NC group were 0.22 ± 0.03 and 0.98 ± 0.06 (F=425.37,P<0.01). The relative expression levels of protein in shRNA-ADAM12 group, shRNA-NC group and shRNA-C group were 28.72%±2.36%, 69.21%±3.92%and 69.04%±3.57%, respectively (F=145.42,P<0.01). Immunofluorescence staining showed that expression levels of ADAM12 and CD133 in cell spheres were significantly higher than those in normal cells. The number of spheres in three groups were 45.5±2.3、104.2±5.8 and 109.6±6.2, tumor sphere formation ability of shRNA-ADAM12 group was lower than that of shRNA-NC group and shRNA-C group (F=147.03,P<0.01). Compared with the shRNA-NC group and shRNA-C group, the protain expression of GFAP and Tuj1 were increased up to 166% and 146% (P<0.01) whereas the protein expression levels of CD133 and HES1 were down-regulated by 54% and 50% (P<0.01). Conclusion Knockdown of ADAM12 may suppress self-renewal ability of CD133 positive glioma cells by inhibiting the Notch pathway activity.

Objective To explore the addictive tendency of internet and its influence factors among adolescents in Longhua District of Shenzhen. Methods A total of 6552 Students were selected from Longhua District of Shenzhen us?ing randomly stratified cluster sampling method. They were inquired about their gender, grade, family type, behaviors of surfing internet, risk behaviors, and internet addiction tendency with a self-report structured questionnaire and the adoles?cent pathological internet use scale (APIUS). Results The prevalence of addictive tendency of internet was 9.0%(590/6552) among adolescents in Longhua District of Shenzhen. Multiple logistic regression analysis indicated that the addic?tive tendency of internet was significantly and positively associated with male (OR=1.32, P<0.05), living in Minzhi area (OR=1.66, P<0.01), secondary school stage (OR=1.62, P<0.01), uncomplete family type (OR=1.49, P<0.01), surfing in cy?bercafé frequently (OR=2.00, P<0.01), compared with surfing 0~2 h/d, surfing 2~4 h/d (OR=1.92,P<0.01), 4~5 h/d (OR=2.97, P<0.01) or more than 6 h/d (OR=4.90, P<0.01), playing games (OR=3.27, P<0.01), chatting (OR=2.80, P<0.01) or having other kinds of entertaining (OR=2.80, P<0.01), and having risky behaviors (OR=2.39, P<0.01). Conclusion The addictive tendency of internet is affected by multiple factors among adolescents in Longhua District of Shenzhen. There?fore, comprehensive intervention measures shall be used to regulate adolescents habits of using internet and prevent them from the internet addiction.

Objective To study the expression changes of IL-1β、FN-α6、IFN-γ and TNFR-m18 in peripheral blood within 2 hours in epileptic mice. Methods Semi quantitative real-time PCR was used to test the mRNA expres?sion level of IL-1β、FN-α6、IFN-γand TNFR-m18 in peripheral blood from normal and pilocarpine-induced epileptic mice at different time points (10 min, 30 min, 1 h and 2 h). Results The mRNA expression level of IL-1βincreased at 30 min(1.8±0.07), 1 h(2.9±0.98)and 2 h(1.45±0.11)after pilocarpine-induced status epilepticus comparing with that of control and SE 10 min(0.81±0.09)(P<0.05). The IFN-α6 mRNA expression level was lower at 10 min(0.59±0.05, P<0.05) than that of control. IFN-γmRNA expression level was higher at 10 min(2.85±0.11) than that of control and at oth?er time points during SE(P<0.01). TNFR-m18 mRNA expression level was higher at 1h(2.84±0.15) than that of control, and at other time points during SE(P<0.01). Conclusion The immune system of epileptic state is active, the imbalance of cytokine expression in peripheral blood may be related to the immune pathological process of acute stage of epilepsy.

Objective To investigate the clinical manifestation, inherited pattern and the related factor of Hunting?ton disease families. Method The clinical data from 12 HD families was collected from 2013-2014. Patients received the genetic test and neurological evaluation including motor, cognitive and problem of behavior. Results There were 12 patients having the IT15 gene dynamic mutations, including 1 Juvenile Huntington disease patient and 3 pre-symptomat?ic mutant gene carriers. The average CAG repeats of these patients was between the range of 40 to 60, and the average on?set age ranged from 13 to 54 year-old. Positive family history and genetic anticipation could be observed. Patients pre?sented with different clinical manifestations at the early stage while had typical chorea movements, declined cognitive and psychiatric symptoms at the late stage of the illness. Conclusions There are typical triad symptoms in the late stage but not in the early stage nor pre-symptom stage illness. Clinical manifestation and the neuroimaging are both of great ref?erence value, and the genetic test is essential for final diagnosis.

Objective To investigate the hemodynamic parameters to stabilize cerebral blood flow within limits of cerebral autoregualtion. Methods We assessed the flow velocity of middle cerebral artery using transcranial Doppler and recorded invasively the blood pressure simultaneously. We then analyzed the curves of cerebral blood flow autoregulation (CBFA) and calculated upper limit of autoregulation (ULA) and lower limit of autoregulation (LLA). The values of critical closing pressue (CCP) and resistance area product (RAP) were calculated according to previous theory. The relationship between CCP, RAP and MABP were analyzed. Results In the process of increasing or decreasing blood pressure, ULA and LLA of normal rats were 148.12 ± 7.49 mmHg or 62.96 ± 3.34 mmHg, respectively. When mean artery blood pressue (MABP) changed within limits of cerebral autoregulation, the cerebral blood flow velocity changed little (increasing:0.65± 0.27 cm/s/10mmHg MABP, decreasing:0.43±0.23cm/s/10 mmHg MABP), while CCP and RAP changed significantly (in?creasing: 4.60 ± 1.06 mmHg/10mmHg MABP and 0.11 ± 0.04/10 mmHg MABP, decreasing: 6.74 ± 0.59 mmHg/10 mmHg MABP and 0.09 ± 0.02/10mmHg MABP). After fixing change of blood flow velocity, CCP and RAP were correlated with MABP more remarkablely, although all blood flow velocity, CCP and RAP were significantly correlated with MABP. Conclusion Within limits of cerebral autoregulation, stable cerebral blood flow is mainly achieved by the change of CCP and RAP against blood pressure changing on normal rats, especially the increasing or de?creasing of CCP.

Objective We investigated the expression profile of cancer related genes in hMSCs co-cultured with U251 glioma cells, to evaluate the risk of malignant transformation of hMSCs in glioma environment. Methods hMSCs were co-cultured with U251 glioma cells for 5 days and the expression profile of cancer-related genes were investigated by using microarray assay, followed by Real-time quantitative RT-PCR and Western blot. Results Of the 440 cancer-re?lated genes covered by Oligo GEArray Human Cancer Microarray OHS-802, SPINT2, TK1, STC1, MMP1, CCND1, SORT1, SEPT6, CDC20, SHB, CDK5, RELA, XRCC4, KIT, CTPS, CAPNS1 and ETV6 were significantly upregulated (>3-fold) whereas none was downregulated in hMSCs co-cultured with U251 glioma cells. The upregulation of oncogenes KIT, CAPNS1, TK1, MMP1, CCND1, CDC20, RELA and STC1 in co-cultured hMSCs were confirmed by Real-time quan? titative RT-PCR. The upregulation of protein expression of oncogenes KIT, MMP1, CCND1 and RELA were detected by Western blot. Conclusion The present study demonstrates that co-culture of hMSCs with human glioma cells leads to up?regulation of some important oncogenes in hMSCs, indicating the tumorigenic potential of hMSCs in glioma environment.

Objective To evaluate the reliability and validity of a medication dependence questionnaire in mi?graineurs of the Chinese version. Methods One hundred fifteen migraine patients with drug dependence syndrome were included and their clinical data were then collected. All the subjects were interviewed with the medication dependence questionnaire in migraineurs of the Chinese version. The reliability and validity of the questionnaire were evaluated ac?cording to test-retest reliability and internal consistency reliability, and the content validity and construct validity respec?tively. Results The test-retest reliability of the questionnaire items was 0.725~1.000 except item 11 (0.558) and item16 (0.443). The internal consistency coefficient of the questionnaire was 0.820, internal consistency coefficient of each di?mension was above 0.7. Every item score was positively correlated with corresponding dimension score (r=0.595~0.962). Seven factors with latent roots greater than 1 could explain 76.3%of total variance, and these factors were relatively pow?erful in related items. Conclusions The medication dependence questionnaire in migraineurs of the Chinese version shows preferable reliability and validity. Thus, this is a promising assessment tool to evaluate the medication dependence in Chinese migraineurs in future clinical practice.

Objective To Compare characteristics between the psychological aids hotline callers who had single attempted suicide and those callers who had repeated attempted suicide. Methods The analysis was conducted on the da?tabase of Beijing psychological aids hotline from Dec., 2002 to Dec., 2008. All the effective callers were consulted by pro?fessional hotline operators. During the calling, the callers were interviewed on their attempted suicides, depression and 11 correlates associated with caller's suicidal behaviors. Results Among 4519 callers who had attempted suicide, 2441 (54%) of them had attempted suicide once, and the other 2078 (46.0%) callers had repeated suicide attempts. After ad?justed for gender, age, and the other 4 demographic variables, history of being abused (OR=1.35, 95%CI: 1.08~1.67), afraid of being attack (OR=1.35, 95%CI: 1.01~1.59), relatives or associates with prior suicidal behavior history (OR=1.17, 95%CI: 1.001~1.36), hopelessness (OR=1.20, 95%CI: 1.02~1.40), and psychological treatment history (OR=0.73, 95%CI:0.62~0.86) were associated with repeated suicide among hotline callers. Conclusions To improve the effective?ness of hotline based suicide prevention, assessment of risk of repeated attempted suicide among the callers with a histo?ry of prior attempted suicide should focus on the history of being abused, afraid of being attack, hopelessness, relatives or associates with prior suicide history, and psychological treatment history.

Objective To investigate the risk factors for intracranial infection after external ventricular drainage and provide basis for preventing and controlling the drainage-associated intracranial infection. Metheds the clinical data from three hundred sixty-seven cases of ventricular hemorrhage patients were retrospectively analyzed, using Logis?tic regression to screen risk factors of intracranial infection after external ventricular drainage. Results There were 29 cases with intracranial infection and infection rate was 8.19%, 8.04% and 7.32% at ventricle drainage tube indwelling 1-week group, 2-week group and 3 week-group, respectively. Glasgow coma score (GCS) [OR= 2. 569 CI (1.792 3.378) %, P< 0.05), urokinase perfusion (OR= 2.897, 95%CI (1.297 5.061), P< 0.05), cerebrospinal fluid sampling (OR= 3.399, 95%CI (2.705 4.175), P< 0.01] and comorbidities [OR= 3.751, 95%CI (2.032 5.371), P< 0.01] were risk factors for ventricle drainage operation. Conclusion Ventricle drainage tube indwelling 3 weeks is safe. Less use of urokinase perfusion and cerebrospinal fluid sampling and active treatment of comorbidities diseases can reduce the intra?cranial infection incidence of external ventricular drainage after Intraventricular hemorrhage .

Objective To analyze the clinical features of rapid eye movement sleep behavior disorder (RBD) in pa?tients with Parkinson’s disease (PD) and investigate correlative factors of RBD. Methods Sixty-three consecutive PD pa?tients were included and classified into PD+RBD group (n=28) and PD-RBD group (n=35) according to REM Sleep Be?havior Disorder Questionnaire (RBDQ-HK). The degree of motor symptoms was compared using Unified Parkinson Dis?ease Rating Scale (UPDRS) and Hoehn&Yahr (H-Y) grade, the incidence of non motor symptoms was compared using non motor symptoms questionnaire (NMSquest), and the cognitive function, anxiety, depression, daytime sleep were com?pared using Montreal Cognitive Assessment (MOCA), Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI), Epworth Sleep Scale (ESS) between the two groups. Results The incidence of RBD in PD patients was 44.4% (28/63). There were longer illness course ( χ2=12.733, P=0.002), older age (t=-2.292, P=0.025), and higher H-Y grades (χ2=7.014, P=0.008) in PD+RBD group, compared with those in PD-RBD group, but there were no significant differences in sex, onset age, onset form and levodapa dose equivalents (LDE) between the two groups. There were higher UPDRSⅡ,Ⅲ scores (t=-2.734, P=0.008; U=3.329, P=0.001) in PD+RBD group. Most of the non motor symptoms, including the gastrointestinal dysfunctions, psychiatric comorbidity and sleep disturbances were more frequent in PD+RBD group (P<0.05), and the incidence of anxiety and depression were higher (χ2=3.958, P=0.047; χ2=10.338, P=0.001), whereas there were no significant differences in cognitive function and daytime sleep between the two groups. In addition, constipation (OR=7.257), illness course (OR=5.389), UPDRS Ⅲ scores (OR=1.060) were correlative with RBD in PD patients. Con?clusion PD patients with longer illness course, older ages, and severe motor and non motor symptoms more likely suffer from RBD. Besides, constipation, long illness course and high UPDRSⅢscores may be risk factors of RBD.