Forensic Sciences/education*

Objective To assess the effects ofberberine on autophagy in C3H10T1/2 adipocytes.Methods C3H10T1/2 cells,a pluripotent stem-cell line of mesodermal origin,were induced to differentiate into mature adipocytes,and then were treated with berberine,The expression of autophagy marker protein LC3 Ⅱ/Ⅰ and autophagy substrate P62 was determined by Western blot ; after treating C3 H 10T1/2 cells with berberine and lysosomal inhibitor,chloroquine,autophagy flux was assessed by Western blot.Autophagosome was observed by transmission electron microscopy after berberine treatment.Results (1) After berberine treatment,the expression of LC3 Ⅱ / Ⅰin C3H10T1/2 adipocytes was reduced and P62 was increased in a time-and dose-dependent manner(P＜0.05 or P＜0.01) ; (2) Following treatment with chloroquine and berberine,the protein level of LC3 Ⅱ/Ⅰ was decreased (P＜0.05); (3) The number of autophagosome was decreased apparently after berberine treatment.Conclusions Berberine inhibits autophagy in C3H10T1/2 adipocytes.

Objective To compare the expressions of Beclin 1 and its 11 interacting proteins in subcutaneous fat and visceral fat,and to observe the expressions of these genes in 3T3-L1 adipocytes differentiation process.Methods In 3T3-L1 adipocytes,cells were harvested at day 0,2,4,6,8 in differentiation process,and then proteins or mRNAs were obtained and followed with Western blot and realtime PCR.Mice tissue proteins or mRNAs were detected with Western blot and realtime PCR.Results In 3T3-L1 adipocytes differentiation process,expressions of LC3,Beclin 1,and TNFAIP3 continued to rise,expression of BIRC5 was high to low,FEZ1 continued at a low level.In mice,expressions of LC3,Beclin 1,YWHAQ,FEZI,BAD,WAC,TNFAIP3,and GOPC in visceral fat were significantly higher than those in subcutaneous fat,while expression of SLAMF1was higher in subcutaneous fat.Conclusion Autophagy plays an important role in 3T3-L1 differentiation process and may be tissue-specific in visceral fat and subcutaneous fat.

0.05) ;incidence of aleucocytosis was 43.3% vs.27.5%(P

Objective To investigate the effects of Tiaoxin Formula on spatial learning and memory ability and long-term potentation (LTP) in the hippocampus of APP/PS1 transgenic mice with Alzheimer disease; To discuss its mechanism of action. Methods Totally 54 three-month-old APP/PS1 transgenic mice were randomly divided into model control group, Tiaoxin Formula group and positive control group, with 18 mice in each group. Another 18 three-month-old C57BL/6J wild mice were chosen as normal control group. All administration groups receive relevant medicine. 12 weeks later, Morris water maze test was used to test behavior and in vitro electrophysiology record. Results The Morris water maze test showed that in place navigation test, compared with the model control group, escape latency time in Tiaoxin Formula group was significantly reduced (P<0.01). In the space experiments, compared with the model control group, the number of crossing the platform quadrant and the time staying at the platform quadrant in Tiaoxin Formula group were significantly increased (P<0.05, P<0.01). In vitro hippocampal electrophysiological induction, compared with the model control group, fEPSP slope in the hippocampus in Tiaoxin Formula group were significantly increased (P<0.01), the PPF had no significant difference (P>0.05). Conclusion Tiaoxin Formula can improve the spatial learning and memory ability and LTP of APP/PS1 transgenic mice with Alzheimer disease, thus can realize cognitive protection effects.

AIM: To observe the morphologic changes of of filtration blebs after trichostatin A treatment in an experimental glaucoma filtration surgery ( GFS) .
METHODS:Subconjunctival injection TSA, mitomycin C ( MMC) and PBS during the filtering surgery in rabbits. The morphologic changes of filtration blebs were evaluated by Krofeld score method postoperatively days 3, 7, 14, 21, and 28.
RESULTS: TSA induced filteation bleds were elevated diffusely within 14d and cystic blebs formed 28d, filtration bleb score was significantly higher in TSA group than that in PBS group.
CONCLUSION: TSA can keep the aqueous humor outflow by inhibiting scar formation and prolong the existence of the filtration bleb.

Objective To investigate the effects of puerarin on the expression of human umbilical vein endothelial cells (HUVECs) tissue factor (TF) and tissue factor pathway inhibitor (TFPI) induced by oxidized low-density lipoprotein (ox-LDL).Methods After HUVECs were incubated with different concentrations of puerarin and 50 mg/L ox-LDL,the expression of TF and TFPI mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blot respectively.Results Compared with control,treatment with ox-LDL caused the augment of TF mRNA and protein expression (P<0.01),and the decrease of TFPI mRNA and protein expression.However,50,100,and 200 μmol/L puerarin blunted the augment of TF mRNA and protein expression and weakened the inhibition of TFPI mRNA and protein expression induced by ox-LDL(P<0.01).Conclusions Puerarin reduces HUVECs TF and TFPI mRNA and protein induced by ox-LDL.

Objective To explore the relationship of nitricoxide synthase(NOS)and cell apoptosis in epilepsy.Methods Pentylenetetrazol(PTZ)was used to build models of epilepsy.Cell apoptosis in each group were detected by flow cytometer,level of NOS was detected by colorimetric method.Results Levels of NOS increased obviously 1 hour after epilepsy and decreased;but at the 6th hour,NOS increased again and peaked at the 24th hour.Cell apotosis begin to increase at 6th hour and peaked at 24th hour after epilepsy,and decreased at 48th hour after epilepsy,there were significance differences compared with control group(Pa

AIM: To explore the role of phosphatidylinositiol 3-kinase/protein kinase B/endothelial nitric oxide synthase (PI3K/Akt/eNOS) signaling pathways in the inhibitory effects of puerarin on oxidized low-density lipoprotein (ox-LDL)-induced tissue factor (TF) expression in vascular endothelial cells.METHODS: The mRNA expression of TF was detected by real-time fluorescent quantitative PCR.The protein levels of TF and Akt was determined by Western blot.The content of the nitric oxide (NO) was measured by nitrate reduction method.RESULTS: Compared with control group, incubating endothelial cells with ox-LDL significantly induced TF expression at mRNA and protein levels and the dephosphorylation of Akt protein, and decreased NO production.Incubation of the endothelial cells with puerarin for 1 h and then treatment of the cells with ox-LDL decreased the TF expression at mRNA and protein levels, increased Akt protein phosphorylation and intracellular NO content.Co-incubation of the endothelial cells with PI3K inhibitor LY294002 and puerarin for 1 h and then treatment of the cells with ox-LDL augmented the TF expression at mRNA and protein levels and the Akt protein dephosphorylation, and decreased NO production.Co-incubation of the endothelial cells with eNOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) and puerarin significantly decreased the inhibitory effect of puerarin on ox-LDL-induced TF expression at mRNA and protein levels in the endothelial cells, and reduced Akt protein phosphorylation and NO production.CONCLUSION: Puerarin inhibits ox-LDL-induced TF expression at mRNA and protein levels in the human umbilical vein endothelial cells via activation of PI3K/Akt/eNOS signaling pathway.

Objective To observe the changes of cAMP response element binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway in the hippocampus of APP/PS1 double transgenic Alzheimer's disease mice, and in-vestigate the effects of Tiaoxin formula on learning and memory. Methods Totally, 54 three-month-old APP/PS1 transgenic mice were randomly divided into model group (n=18), posi-tive control (donepezil) group (n=18) and Tiaoxin formula group (n=18). Another 18 three-month-old C57/BL6 wild mice were chosen as normal group with same strain and same age. All administration groups received rele-vant medicine. Twelve weeks later, Morris water maze test was used to test the behavior. The real-time quantita-tive polymerase chain reaction (RT-qPCR) was used to detect the expression of CREB mRNA and BDNF mRNA in hippocampus. Western blotting was used to detect the expression of phosphorylated CREB (p- CREB) and BDNF in hippocampus. Results Compared with the model group, the escape latency significantly shortened (P<0.01), the percentage of staying time in the quadrant of original platform significantly increased (P<0.05), the expression of CREB mRNA and BDNF mRNA in hippocampus significantly increased (P<0.05), and the expression of p-CREB and BDNF pro-tein in hippocampus significantly increased (P<0.01) in Tiaoxin formula group. Conclusion Tiaoxin formula could improve the spatial learning and memory of APP/PS1 double transgenic mice with AD, which might be related to the up-regulation of CREB/BDNF signaling pathway.