Objective: To study the effects of glycosaminoglycan from scallop skirt (SS-GAG) on the formation of foam cells from porcine artery smooth muscle cells (SMCs) and the expression of the total superoxide dismutase(SOD)and glutathione peroxidase(GSH-PX)activity, NO production and the mechanism of anti-atherosclerosis action of SS-GAG. Methods: SMCs were incubated with 15 mg/L oxidized low-density lipoprotein (Ox-LDL) for 72 h to establish a smooth muscle cell-derived foam cell model. In addition, SMC cells were divided into 6 groups:①blank group,②model(Ox-LDL) group, ③Ox-LDL+200 mg/L SS-GAG group,④Ox-LDL+400 mg/L SS-GAG group,⑤Ox-LDL+800 mg/L SS-GAG group, ⑥Ox-LDL+Heparin 100 mg/L group. After 72 h incubation, intracellular total cholesterol (TC), free cholesterol (FC), cholesteryl ester (CE) content and CE/TC ratio were measured through enzymatic method. The SOD, GSH-PX and NO concentration in the medium were also determined through Xanthine oxidase method or TBARs. Results: TC, CE, CE/TC in model group significantly increased, while FC, GSH-PX and NO concentration in the medium significantly decreased compared with blank group. After treatment with heparin (100 mg/L) and different concentrations of SS-GAG (200 mg/L, 400 mg/L,800 mg/L), TC, CE, and CE/TC significantly decreased (P

Drugs, Chinese Herbal ; therapeutic use ; Humans

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 dynamic changes in microvascular ultrastructure in the cortex after the acute cerebral ischemia-reperfusion.Methods40 male rats were randomly divided into two groups（n=20 for sham operation group and cerebral ischemia-reperfusion group）. Cerebral ischemia-reperfusion model was produced using suture middle cerebral artery occlusion. Rats were sacrificed and the brain samples were adopted 1,3,12,72 h after ischemia-reperfusion, methyl methacrylate composite brain microvascular casting. The production of brain microvascular specimens, scanning electron microscopy of normal rat cerebral cortex microvessels and cerebral cortex of acute brain injury morphological changes in microvascular.ResultsCompared with the sham-operated group, cerebral ischemia-reperfusion in the cortex after the signs of vascular damage, then, vascular casting was to "bean" shape or even had a completely broken "tears candles" stump-like vascular casting, finally, to further the formation of a vascular zone cortex. ConclusionThe structural changes of brain microvascular in the cortex after acute cerebral ischemia-reperfusion is an important cause of cerebral microcirculation in rats.

OBJECTIVETo study the effect of single administration of aqueous extracts from aconite on "dose-toxicity" relationship and "time-toxicity" relationship of mice hearts, through changes in electrocardiogram (ECG) and serum biochemical indexes.

METHODMice were grouped according to different drug doses and time points, and orally administered with water extracts from aconite for once to observe the changes of mice ECG before and after the administration, calculate visceral indexes heart, liver and kidney, and detect levels of CK, LDH, BNP and CTn-I in serum.

RESULTAccording to the "time-toxicity" relationship study, at 5 min after oral administration with aqueous extracts from aconite in mice, the heart rate of mice began rising, reached peak at 60 min and then slowly reduced; QRS, R amplitude, T duration and amplitude and QT interval declined at 5 min, reduced to the bottom at 60 min and then gradually elevated. The levels of CK, LDH, BNP and CTn-I in serum elevated at 5 min and reached the peak at 60 min, with no significant change in ratios of organs to body at different time points. On the basis of the "dose-toxicity" relationship, with the increase in single dose of aqueous extracts from aconite, the heart rate of mice. QRS, T duration and amplitude and QT interval declined gradually, and levels of CK, LDH, BNP and CTn-I in serum slowly elevated, with a certain dose dependence and no significant change in ratios of organs to body in mice.

CONCLUSIONSingle oral administration of different doses of aqueous extracts from aconite could cause different degrees of heart injury at different time points, with a certain dose dependence. Its peak time of toxicity is at 60 min after the administration of aqueous extracts from aconite.

Aconitum ; adverse effects ; chemistry ; Animals ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Female ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; Kidney ; drug effects ; Liver ; drug effects ; Male ; Mice

Objective To study the changes of proteome expression in brain tissues from rats with severe traumatic brain injury(sTBI). Methods Total protein of brain tissues were obtained at days 3,7 and 14 for two-dimensional gel electrophoresis to screen and identify differential protein spots.Results We screened 17 differential protein spots that were involved in cellular metabolism,stress and inflammatory reaction. Conclusion Some differential proteins involved in sTBI can be found by twodimensional gel electrophoresis.

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.

Objective To investigate the effect of temperature sensitive umbilical cord mesenchymal stem cells (tsUCMSCs) transplantation under mild hypothermia environment on cognitive function of rats after severe traumatic brain injury (TBI).Methods Thirty-six male SD rats weighing (300 ± 20)g were subjected to severe TBI by fluid percussion device (2.5 atm) and managed with tsUCMSCs combined with mild hypothermia.Rats were divided into three groups:sham operation group,TBI group,and TBI + tsUCMSCs + mild hypothermia group (TBI + intervention group) according the random number table,with 12 rats per group.Latency to the platform and the time in targeted quadrant were recorded to evaluate the learning and memory with Morris water maze.Population spike (PS) and excitatory postsynaptic potential (EPSP) of long-term potential (LTP) were examined to evaluate synaptic plasticity of hippocampus.Phosphorylated Tau at Thr231 and phosphorylated GSK3 at Ser9 were detected by Western blot.Results Latency to the platform [(15.9 ±3.2) s vs (8.08 ±2.69) s,P＜0.01]was prolonged and time in targeted quadrant [(14.4 ± 3.0) s vs (36.3 ± 5.7) s,P ＜ 0.01] was shortened in TBI group at day 28 compared with sham operation group.Whereas,latency to the platform [(10.7 ± 2.8) s,at Ser9 were detected by Western blot.Results Latency to the platform [(15.9 ± 3.2) s vs (8.08 ± 2.69) s,P ＜ 0.01] was prolonged and time in targeted quadrant [(14.4 ± 3.0) svs (36.3 ± 5.7) s,P ＜0.01] was shortened in TBI group at day 28 compared with sham operation group.Whereas,latency to the platform [(10.7 ± 2.8) s,P ＜ 0.01] and time in targeted quadrant [(29.4 ± 4.4) s,P ＜ 0.05 |were reversed in TBI + intervention group.PS and EPSP of LTP were elevated in TBI group,but the elevation was suppressed in TBI + intervention group.Meanwhile,Tau hyperphosphorylation (0.80 ± 1.00vs 1.24 ±0.13,P＜0.05) and GSK3 deactivation (3.01 ±0.41 vs 1.27 ±0.22,P ＜0.01) were significantly reversed in TBI + intervention group compared with TBI group.Conclusion Combination of tsUCMSCs and mild hypothermia therapy can improve the TBI-induced cognitive deficit.

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 judge injury severity of severe traumatic brain injury (sTBI) by using surface enhanced laser desorption-ionization (SELDI) protein chip technique. Methods Serum sam-ples from sTBI patients were used to detect expression of differential proteins by protein chip CM10 and SELDI to analyze the correlation between expression peak intensity and GCS. Results We obtained 101 protein peaks, with statistical difference upon expression of 27 protein peaks, when negative correla-tion was found between two peaks ( m/z 4 972 and m/z 5 322 ) and GCS score and positive correlation be-tween six peaks (m/z 3 941, m/z 4 295, m/z 8 714, m/z 8 792, m/z 14 020 and m/z 28 148) and GCS score. Conclusion SELDI protein chip technique may become a new and objective detection method in judging injury severity of sTBI.