To generate recombinant adenovirus expression vector of human Sema4C gene and observe its expression in mouse myoblasts cell line C2C12 for ensuring easy access to investigate the role of Sema4C gene during myogenesis. The recombinant plasmid was packaged and amplified after being transfected in HEK293 cells through Lipofectamine. After infecting C2C12 myoblasts with recombinant adenovirus vector, the adenoviral infection efficiency was determined by confocal microscope which showed that the expression of green fluorescence could be detected at 12h and then reached peak at 24h after recombinant adenovirus infection. The infection efficiency was almost 100% confirmed by FACS examination. Detection of WB indicated that the expression of Sema4C in C2C12 of recombinant adenoviral infection group was significantly higher than that of the control group (P

AIM:To investigate the protective effects of adenosine on cultured rat hippocampal neurons after oxygen-glucose deprivation. METHODS:The control and adenosine-treated hippocampal neurons cultured for 12 d were exposed to oxygen-glucose deprivation environment for 0.5-4 h and then cultured with original medium in normoxia for 24 h . The soma area,survival rate, effluxes of lactate dehydrogenase (LDH)and apoptosis of neurons were observed. RESULTS:The soma area, effluxes of lactate dehydrogenase from neurons and apoptosis were increased while survival rate of neurons was decreased after oxygen-glucose deprivation compared with those pre-oxygen-glucose deprivation. Compared with the control, after oxygen-glucose deprivation the soma area, effluxes of lactate dehydrogenase from neurons and apoptosis were decreased, however, the survival rate of neurons was increased in the adenosine group.CONCLUSION: Oxygen-glucose deprivation can lead to the severe damage of cultured hippocampal neurons, and adenosine can reduce neuronal injury induced by oxygen-glucose deprivation.

Adolescent ; Adult ; Aged ; Facial Nerve ; anatomy & histology ; pathology ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Parotid Neoplasms ; surgery ; Young Adult

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.

Objective To investigate the effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems of school-aged children in Shanghai. Methods A total of 4 108 school-aged children from 10 primary schools of Shanghai were enrolled by multi-stage cluster sampling and surveyed by questionnaires. The information of television-watching and computer-using, family and personal condition was investigated by self-prepared questionnaire, and the Chinese version of Children's Sleep Habits Questionnaire was employed to survey the sleep behaviors of children. The effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems were analyzed by multiple linear regression analysis and Logistic regression analysis. Results The percentage of children who watched television≥2 h per day was 4.1% during weekdays, and that came to 49.2% during weekends. In terms of frequency of computer-using, most children reported "rarely" (88.2%, 0-1 time/week), followed by "often" (11.0%, 2-4 times/ week) and "usually" (0.8%, 5-7 times/week). With the age increase, the percentages of children who watched television≥2 h per day and those who "often" used computer gradually increased. It was revealed by multiple linear regression analysis and Logistic regression analysis that television-watching and computer-using were not only positively correlated with later bedtime, later wake time and shorter sleep duration but also significantly associated with sleep problems such as bedtime resistance, sleep onset delay, sleep duration disorder, sleep anxiety and parasomnia. Conclusion Television-watching and computer-using exert influences on sleep behaviors of sleep/wake patterns, sleep duration and sleep problems. Concerns about the potential negative effects of television-watching and computer-using on sleep behaviors may help to promote healthy sleep patterns and improve sleep quality.

AIM: To determine the effect of heat treatment on rat primary cultured neurons, and give fundamental research for candidate molecule to protect the neurons from heat injury. METHODS: Neurons from rat striatum were primary cultivated in D-MEM with 15% horse serum, and when got mature, cells were identified by immuno-cytochemical staining with neurofilament protein (NF), tyrosine hydroxylase (TH) and neuron specific enolase (NSE) antibodies. Cells in heat treatment groups were put in an 43 ℃ CO 2 incubator for 1 h, and the control groups at 37 ℃ as normal. Striatum neurons were stained with trypan blue and dual fluorscence dye (PI/H33258) immediately followed heat treatment, and necrosis rate of neurons was estimated. At the same time, activated caspase-3 immuno-cytochemical and TdT TUNEL methods were applied to determine apoptosis rate, and cell volume was also identified with micro-photography. RESULTS: During day 7 to day 9, the cultured striatum neurons got mature, and many neuronal fibers starched out and formed neuron network, NF, TH, and NSE staining positive. Treatment at 43 ℃ for 1 h, cell number decreased greatly, while NF+ percentage kept unchanged, and the heat treatment survived neurons were processing cell necrosis and apoptosis, but necrosis percentage was much greater than that of apoptosis. While cell volume kept unchanged after heat treatment. CONCLUSION: Heat treatment greatly affects the growth and survival of the cultured striatum neurons, and the injury effect is most due to cell necrosis process.

Aim To study the antidepressant effects of bioactive compounds from entomogenous fungi(BCEF) in mouse models of depression. Methods The antidepressant effects of Bioactive compounds from entomogenous fungi was examined on the chronic unpredictable stress test, yohimbine induced lethality test, and 5-HTP induced head-twitches test. Results BCEF(50 mg?kg~-1 , ig, qd?21 d)could significantly increase the crossing and rearing score in open-field test. After administration yohimbine 1 h, BCEF 100 mg?kg~-1 group mice mortality rate rising rapidly; BCEF(50,100 mg?kg~-1 ) could distinctly increase the head-twitch number in the 5-HTP induced head-twitches test. Conclusion BCEF has antidepressant effects in depression mouse models. The mechanisms of its antidepressant effects may be related with the reinforcement of central monoamine neurotransmitter especially to 5-HT and NE nerves system.

Background Chemical crosslinking agent can be used to strengthen the intensity of sclera tissue,but the intensity of the sclera may be influenced by different crosslinking methods.Objective The aim of this study was to compare the effectiveness of collagen crosslinking on porcine sclera between whole-eye crosslinking method and scleral strip crosslinking method.Methods Whole-eye crosslinking or sclera strip crosslinking was performed on 70 fresh porcine eyeballs in five groups using 1％ genipin,1％ glutaraldehyde or PBS respectively for 40 minutes.After crosslinking,10 sclera strips with l0 mm×4 mm from the temporal lateral were prepared in every group for the stress-strain measurement using a Instron5848 microtester,and the other 4 scleral strips in each group were extracted for the thermal shrinkage temperature test.Results Biomechanical property test reveled that the elastic modulus value of sclera strips reduced by 70.0％-82.8％ in the whole-eye crosslinking method group compared with scleral stip crosslinking method group after treated with 1％ genipin ((8.98 ± 1.81) MPa vs.(10.85 ± 1.83) MPa,t =3.375,P =0.003)) and 1％ glutaraldehyde((12.78 ±2.91) MPa vs.(18.25 ±5.16) MPa,t =4.007,P =0.001)) ;The tensile stress of whole-eye crosslinking method group was 54.9％-90.1％ of scleral stips method group,showing significant decline after corsslinked of whole-eye in 5％,10％,15％ and 20％ strain conditions (all P ＜ 0.05).Thermomechanics test showed that the thermal shrinkage temperature was lower in the whole-eye crosslinking group compared with scleral stip crosslinking group after treated with both 1％ genipin ((68.8 ±0.9)℃ vs.(74.8± 1.3)℃,t=11.129,P=0.000)) and 1％ glutaraldehyde((73.3±0.9)℃ vs.(79.3±1.3)℃,t=11.112,P=0.000)).Conclusions Different crosslinking methods have an influence on the efficacy of collagen crosslinking on porcine sclera.Sclera strip crosslinking offers a better crosslinking intensity for selera.

Objective: To explore the effect and mechanism of Longzhu ointment in the treatment of allergic contact dermatitis.Methods: Allergic contact dermatitis mouse model was induced by dinitrobenzene (DNFB) and the mice were randomly divided into the model group, matrix group, Longzhu ointment group and hydrocortisone butyrate group.0.5% DNFB was used for sensitization on abdomen of mice one and two days before the administration, and 0.2% DNFB was used on the right ears of mice after 6-day administration to establish mouse ACD model.A day after the excitation of dermatitis, the ear swelling degree was determined, the changes of histopathology was observed, and the immunohistochemical methods were performed to quantify the expression of TNF-γ and macrophage surface markers CD68 in the tissue samples.Results: The ear swelling degree in the group treated with Longzhu ointment was significantly reduced (P <0.01).Compared with the postive control group,the difference was not statistically significant(P＞0.05).HE staining showed that tissue edema decreased obviously.The immunohistochemistry analysis revealed that the expression of INF-γ in Longzhu ointment group was obviously reduced.Under the microscope with magnification of 200 times, CD68 positive cells in Longzhu ointment group was significantly reduced than those in the model group(P＜0.01).The expression of CD68 positive cells in butyric acid hydrocortisone group was not significantly different from that in Longzhu Ointment group.Conclusion: Longzhu ointment can inhibit allergic contact dermatitis obviously, and the mechanism may be related to the inhibition of lymphocyte secretion INF-γ and the reduction of macrophage infiltration.

Objective To quantify the amplitudes of lung tumor motion during free-breathing using four dimensional computed tomography (4DCT), and seek the characteristics of tumors with large motion. Methods Respiratory-induced tumor motion was analyzed for 44 tumors from 43 patients. All patients un-derwent 4DCT during free-breathing before treatment. Gross tumor volumes (GTV) on ten respiratory phases were contoured by the same doctor. The eentroids of GTVs were autoplaeed with treatment software (ADAC Pinnacle 7.4f), then the amplitudes of tumor motion were assessed. The various clinical and anatomic fac-tors associated with GTV motion were analyzed. The characteristics of tumors with motion greater than 5 mm in any direction were explored. Results The tumor motion was found to be associated with T stage, GTV size, the superior-inferior (SI) tumor location in the lung, and the attachment to rigid structures such as the chest wall, vertebrae or mediastinum. The motion over 5 mm was observed in ten tumors, which were all lo-cated in the lower or posterior half of the lung, with the greatest motion of 14.4 mm. For 95% of the tumors, the magnitude of motion was less than I 1.8 mm, 4.6 mm and 2.7 mm along the SI, anterior-poste-rior (AP) and lateral directions, respectively. Conclusions Tumor motion due to breathing is associated with tumor location, volume, and T stage. The greatest motion was in the SI direction for unfixed tumor in lower-lobe, followed by tumor in upper-lobe posterior-segment.