Objective To study the effect of brain-derived neurotrophic factor (BDNF) on the environmental nutrition and neural differentiation of the transplanted stem cells under hypothermia.Methods The BDNF gene mediated by liposome was transfected into 293T cell line, and ELISA assay was applied to find the peak time of BDNF expression. When BDNF was highly expressed, the supernatant was collected for establishment of SD rat models of brain injury. The rats were divided into Group A (stem cell transplantation group) and Group B (stem cell transplantation and BDNF group). Rats in both groups were under hypothermia treatment for five days. Four and eight days later ( three days from rewarming), rat brain tissues were obtained to detect the expressions of proliferating cell nuclear antigen (PCNA), nestin, neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunohistochemical method and to detect the apoptosis by in situ hybridization. Finally, the nerve function scores were obtained for evaluation of the nerve function. Results The ELISA showed that the high level of BDNF expression was at 48 to 60 hours after gene transfection. PCNA and nestin were highly expressed, while NES and GFAP showed nil or low level of expression in both groups at the fourth day after hypothermia, with little apoptotic cells especially in the Group B (P ＜0.05). The expressions of PCNA and nestin were decreased, but the expressions of NSE and GFAP were increased at the third day after rewarming. The positive rate of NSE expression in the Group B was much higher and the apoptotic cells were much less compared with the Group A ( P ＜ 0. 05 ). A better nerve score was obtained in the Group B. Conclusion BDNF can enhance the survival rate of the transplanted stem cells and induce their differentiation into neurons under hypothermia.

ObjectiveTo investigate the roles of 11 β-hydroxysteroid dehydrogenase type 1 ( 11 β-HSD1 )on hippocampus of rat with chronic unpredictable mild stress (CUMS).MethodsTwenty-four male SpragueDawley rats were randomly divided into control group and depressive model group. Chronic unpredictable mild stress (CUMS) was used to make up depressive animal model.Behavioral changes were recorded by body weight measuring,sucrose consumption test (SCT) and open field test (OFT),respectively.The mRNA transcription of 11β-HSD1 in hippocampus tissues of the rats were detected by real-time RT-PCR,and the protein expression of 11β-HSD1 were detected by western blot and immunofluorescence.ResultsBcforc starting CUMS protocol,the rats exhibited equivalent weight and sucrose consumption.Twenty-eight days after CUMS protocol,behavior parameters such as body weight,sucrose consumption,nunber of crossing,and number of rearing were significantly decreased in rats exposed to CUMS group compared with control group (P ＜ 0.05,P ＜ 0.01 ).Correspondingly,realtime RT-PCR assays showed the mRNA expression of 11 β-HSD1 in the hippocampus of CUMS group,which was (31 ±9) ％ lower than that of control group.Meanwhile,the protein expression of it in CUMS group was lower than that of control group (P ＜ 0.05 ).Inmunofluorescence revealed that the number of positive 11 3-HSD1 cells was high (223 ± 13) in the control group,while the number was decreased prominently (92 ± 11 ) in the CUMS group (P ＜ 0.01 ).ConclusionDepressive behavior of rats is induced and the expression of 11 β-HSD1 in the hippocampus is decreased prominently by CUMS,the mechanism of which is at least related to the low expression of 11β-HSD1 and disturbance of glucocorticoid metabolism caused by CUMS.

Objective To establish the electric controlled cortical impact (eCCI)-induced traumatic brain injury (TBI) model in rats with different severity in degree,which may serve as a suitable platform to provide experimental evidence for the pathophysiological following TBI.Methods A total of 40 male Wistar rats were randomly divided into 3 experimental groups and sham group.TBI rats (n=10/group) were positioned beneath the controlled cortical impactor device (eCCI) and subjected to impact injury at 2 mm depth of penetration,for a sustained depression of 200 ms,at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.Sham-operated rats (n=10) underwent identical surgical procedures,including craniotomy,without receiving the cortical impact.Neurological function and regional cerebral flow (24 h after CCI),contusion volume,histopathological,and ultrastructural changes (48 h after CCI) were measured,respectively.Results The severity of the pathological changes in rats was increased as the injury aggravated.The eCCI device impacted the brain at 4 m/s,5 m/s,6 m/s velocity for mild,moderate,and severe TBI,respectively.TBI groups showed impaired neurological function,and decreased rCBF lower than that of sham-operated group (all P＜0.01).Furthermore,neuronal pathological abnormalities in TBI groups,including neuron shrinking,perineuronal vacuole,and structural abnormalities of mitochondria.Increased severity of injury was apparent following the increased level of the impacted velocity,and significant differences were observed between TBI groups (P＜0.05).Conclusion The TBI animal model with mild,moderate,and severe brain injury can be established successfully by 4 m/s,5 m/s,and 6 m/s of impact velocity respectively with the eCCI-6.3 device.The novel eCCI-induced TBI model in rats possibly serves as a novel useful approach in the development of TBI models.

The refractory epilepsy refers to the epilepsy whose seizures couldn't be cured after using two kinds of correctly selected antiepileptic drugs which can be tolerated and enough dosage and duration of monotherapy or combination therapy.The pathogenesis of intractable epilepsy is complex,and there is no established theory at home and abroad.Recently,more and more attention has been paid to the correlation between mitochondrial dysfunction caused by oxidative stress and intractable epilepsy.Based on the summary of common treatment of refractory epilepsy and by searching the related literature at home and abroad,further investigation would be made to explore the diagnosis and treatment of intractable epilepsy theory in order to provide new strategies for diagnosis and treatment of intractable epilepsy.

To cultivate medical talents with innovative ability is necessary for quality education,medical development and medical mode transformation.The traditional education mode cannot acclimatize itself to innovation education.So,we should renovate the education concept,train students in innovative ability and put quality education into practice.

Objective To screen altered proteins of hippocampus in the stress-induced depression (STRID) rat model, and explore the potential molecular mechanism. Methods Twenty Sprague-Dawley rats were randomly divided into the control group and STRID group, 10 rats in each group. Chronic unpredictable mild stress (CUMS) methods including fasting for solids and liquids, electric foot-shock, reversing day and night, cold water swimming, cage tilt, scare stimulation and tail pinch were conducted on STRID rats with no repeats for 28 days to make up the depression animal model. The control group was normally fed during this period. After the stress stimulation, the hippocampus protein samples were used for two dimensional electrophoresis to screen the differentially expressed protein, and then mass spectrum identification and function analyze were conducted. Results Compared with the control group, 34 proteins were altered in STRID group. Among which, 18 were up-regulated, and 16 were down-regulated. The differentially expressed proteins mainly located in cytoplasm, mitochondrion, extracellular exosome and myelin sheath. The involved signaling pathways included metabolic pathway, oxidative phosphorylation pathway, and Alzheimer's disease, Parkinson's disease and Huntington's disease pathways. Conclusion The altered proteins and dysfunction of nerve signaling, and the excess of oxidative phosphorylation in hippocampus of STRID rats may be one of the pathogenesises.

Objective To study intermediate and long term efficacy of uterine arterial embolization (UAE)with sodium alginate microspheres(KMG)at diameters 500-700μm in treatment of diffuse adenomyosis.Methods Totally 40 patients with standard difluse adenomyosis were enrolled and treated with UAE.KMG at diameters 500-700 μm for vascular embolization were used to embolize the arteries.The degree of dysmenorrhea,amount of menorrhea and uterine volume,as well as the level of serum CA125,follicle stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E2)were investigated before andafter UAE.Results The follow up rates were 100%(40/40),100%(40/40),80%(32/40),68%(27/40),58%(23/40)after uterine arterial UAE 12,24,36,48 and 60 months respectively.The early,intermediate and long-term effective rates were 90%(36/40),88%(28/32),83%(19/23).The degree of dysmenorrhea,the amount of menorrhea and the uterine volume,as well as serum CA125 all decreased significantly 3 mouths after UAE at varying degrees(P＜0.05).Compared with other follow-up time,thedegree of dysmenorrhea and the amount of menorrhea declined to their lowest point at 6 month after UAE (P＜0.01).Paralleled with the decrease of volume of uterine,serum CA125 also decreased significantly and reached the lowest level 12 months later compared with other follow-up times(P＜0.01).Even at the 12th month after UAE serum CA125was not normal and FSH,LH and E2 did not change all the times after UAE(P>0.05).No recurrence was found during the 60 months after UAE.Condusion KMG used in UAE at diameters 500-700 μm has good intermediate and long term effectiveness in treatment of diffuse adenomyosis with no side effects.

Objective To investigate the therapeutic effect of Xuebijing injection on stroke associated pneu monia the changes of plasma C-reactive protein level .Methods 80 cases of post-stroke patients with pneumonia were randomly divided into Xuebijing injection treatment group 40 cases and control group of 40 cases, two groups were given conventional antibiotics and anti-inflammatory treatment ,treated with Xuebijing injection group was dealed with Xuebijing injection 50ml plus 0.9% sodium chloride solution 100ml intravenous drip on the basis of conventional therapy,2/d,for 7d,changes before and after treatment in the two groups were evaluated the body temperature ,periph-eral white blood cell count ,neutrophil percentage ,C-reactive protein index .Results The treatment group after treat-ment for 7d body temperature,blood routine,neutrophils and C-reactive protein were compared before treatment were significantly improved(t=9.99,24.09,12.44,43.98;all P<0.05),all of the indexes in the control group compared before treatment were significantly improved,the differences were significant(t=15.95,20.12,4.14,16.53;all P<0.05),after treatment the observation index except temperatrue decreased significantly ,with statistically significant differences compared with control group (t=4.83,6.15,7.93,all P<0.05).Conclusion Xuebijing injection syner-gistic effect of stroke-associated pneumonia antibiotic treatment significantly , more effective than antibiotic therapy alone,has the very good application and promotion of clinical value .

Objective To investigate the role of three-dimensional scaffolds seeded with NeuroD1-modified neural stem cells (NSCs) for repair of spinal cord injury in rats.Methods A new three-dimensional bio-printer was used to make bionic spinal cord scaffolds.NSCs are transduced with retrovirus vectors encoding NeuroD1 to express transgenes in high levels.Forty healthy female SD rats were divided into control group,scaffold group,scaffold + NSCs-green fluorescent protein (GFP) group and scaffold + NSCs-NeuroD1 group with 10 rats per group,according to the random number table.Spinal cord injury in rats was induced using the electric controlled cortical impactor.A week later,the control group was excised 3 mm spinal cord at the injury site under microscope.RT-PCR was used to confirm the construction of NeuroD1 overexpressing NSCs.Survival and differentiation of transplanted NSCs were detected with fluorescent staining.Rat neurological motor function was evaluated with BBB score at postoperative 1,2,4,6 and 8 weeks.Rat electrophysiological changes were observed by monitoring motion evoked potential and sensory evoked potential at 8 weeks.Results RT-PCR results confirmed the successful reconstruction of NeuroD1-overexpressing NSCs.BBB score in scaffold + NSCs-NeuroD1 group was the highest and had significant differences compared to other three groups (P ＜ 0.05).Electrophysiological results showed the motor and sensory in scaffold + NSCs-NeuroD1 group had the shortest latencies and highest amplitudes,which revealed significant differences compared to other three groups (P ＜0.05).Immunofluorescence staining showed GFP cells in scaffold + NSCs-NeuroD1 group at 8 weeks,which differentiated into neurons and astrocytes.GAP-43 was positively stained,and myelin formation was detected.Conclusion Three-dimensional scaffolds seeded with NeuroD1-modified NSCs can promote nerve loop reconstruction in spinal cord injury rats,and accelerate recovery of motor and sensory function.

Objective To investigate the effects of synuclein-γ (SNCG) gene silencing on the proliferation and apoptosis of glioma U87-MG cells.Methods Five small hairpin RNA templates targeting SNCG and a negative control were synthesized and cloned into the lentiviral vector system and all the constructs were sequenced.Then the recombinant lentiviral vectors were used to infect U87-MG cells.The lentiviruses which can effectively inhibit protein expression levels of SNCG were selected by RT-PCR for further study.Colony formation and flow cytometry assay were used to investigate the effects of SNCG downregulation by RNA interference on the clony formation,proliferation,and apoptosis of U87-MG cells,respectively.Results The lentiviral vectors carrying 5 shRNAs targeting the SNCG gene were successfully constructed,and SNCG siRNA3 and siRNA5 showed higher interfering efficiency than other vectors.In comparison with the group of negative control,SNCG siRNA3 and siRNA5 were observed to significantly inhibit SNCG expression at the mRNA levels (the relative mRNA levels:siRNA3 (0.17± 0.01)％,siRNA5 (0.13±0.01)％ vs (1.00±0.10)％,P＜0.05).Also,SNCG suppression mediated by RNAi significantly inhibited the clone formation (colony number:siRNA3 (66± 12),siRNA5 (1 ± 1) vs (80± 5),P＜0.05),and the proliferation (ratio of cells in S phase:siRNA3 (41.2±0.7) ％,siRNA5 (39.9±0.5) ％ vs (47.6±2.2) ％,P ＜0.05),but promoted the apoptosis (cell apoptosis:siRNA3 (22.9± 0.4) ％,siRNA5 (28.6± 0.9) ％ vs (1.1 ± 0.1) ％,P＜0.01) of transfected U87-MG cells.Conclusion SNCG suppression at the mRNA level mediated by RNAi can inhibit the proliferation and the clony formation,but induce the apoptosis of glioma U87-MG cells in vitro,suggesting that SNCG suppression mediated by an RNAi strategy may become a novel approach for treating human gliomas.