Objective To investigate the effects of minocycline on cell viability and neurite outgrowth of pheochromocytoma cells (PC12) after oxygen‐glucose deprivation(OGD) injury .Methods PC12 cells were exposed to OGD insult for 2 ,4 ,6 ,8 h to estab‐lish a cerebral ischemia model in vitro .High‐differentiated PC12 cells were cultivated and randomly divided into three groups :con‐trol group ,OGD group and various doses of minocycline(0 .1 ,1 .0 ,10 .0 μM) treated group .24 h after OGD‐reperfusion ,PC12 cells viability was assessed by CCK‐8 assay ,the neurite was labeled with MAP‐2 by immunofluorescence and neurite length was meas‐ured by the Image‐Pro Plus 7 .0 software ,GAP‐43 protein expression was determined by Western blotting .Results Compared to the OGD groups ,minocycline induced a concentration‐dependent increase in cells viability [(46 .1 ± 2 .9)% vs .(77 .0 ± 2 .5)% ,P<0.01],improvedneuriteoutgrowthandincreasedtheexpressionofGAP‐43proteininPC12cellsafterOGDinjury([(0.34±0.04) vs .(2 .11 ± 0 .10) ,P<0 .01] .Conclusion Minocycline could protect against oxygen glucose deprivation injury and promote neurite outgrowth .This finding suggests minocycline may be a novel therapy for cerebral ischemia .

Objective To investigate the effects of minocycline on regional cerebral blood flow and the expression of endothe-lin-1(ET-1) in ischemic penumbra of rats after focal cerebral ischemia reperfusion injury .Methods Middle cerebral artery occlusion (MCAO) with nylon suture was used to be established as focal cerebral ischemia reperfusion mode (I/R) ,a total of 35 male Spra-geue-Dawley rats were randomly divided into 3 groups :sham-operated group(n= 10) ,model group(n= 15) and minocycline group (n= 15) .After 24 hours of I/R ,The neurobehavioral function of rats was evaluated by Longa′s test ,the regional cerebral blood flow in ischemic penumbra was assessed with laser-Doppler flowmetry .After 6 and 24 hours of I/R ,the expression of ET-1 in peri-in-farct region was measured by both immunohistochemistry and radioimmunoassay .Results Compared with sham-group ,the Longa′s test scales ,ET-1 protein expression increased and the rCBF decreased in ischemic penumbra in model group(P< 0 .05) .the Longa′s test scales ,ET-1 protein expression decreased and the rCBF decreased in ischemic penumbra in minocycline group when compared to the model group(P< 0 .05) .Conclusion Minocycline could promote neurological functional recovery of rats after MCAO ,which might be attributed to increase the cerebral blood flow and regulate the endothelin-1 expression in ischemic penumbra .

To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.

Objective To investigate the role of poly(ADP-ribose) polymerase (PARP) inhibitor PJ34 in regulating blood-brain barrier (BBB) permeability and matrix metalloproteinases-9 (MMP-9) expression in a mouse model of traumatic brain injury (TBI).Methods A total of 136 adult male BALB/c mice were randomly divided into sham-operated group,injured group and PJ34-treated group according to the random number table.Controlled cortical impact in mice was established.At 6 and 24hours postinjury,neurological deficit was evaluated,including motor,sensory,reflex and beam balance tests ; BBB permeability and brain water content were detected using Evans blue test and gravimetric technique; brain contusion volume was measured using HE staining; levels of MMP-9 in cytosolic fractions were detected using Western blotting.Results At 6 and 24 hours postinjury,neurological severity score in PJ34-treated group (8.00 ± 0.26,7.50 ±0.25) were lower than those in injured group (12.50 ±0.39,11.80 ± 0.32) ; brain contusion volume in PJ34-treated group [(11.25 ± 0.91) mm3,(13.55 ±1.06) mm3] was lower than those in injured group [(25.37 ± 1.75) mm3,(28.24 ± 1.51) mm3] ; BBB permeability in PJ34-treated group [(440.08 ± 3.10) μg/mg,(860.46 ± 3.86) μg/mg] was lower than those in injured group [(936.96 ± 4.71) μg/mg,(1 302.23 ± 5.89) μg/mg] (all P ＜ 0.01).Brain water content lowered significantly in PJ34-treated group than in injured group at 6 hours postinjury [(80.77 ± 0.76) ％ vs (82.55 ± 0.73) ％,P ＜ 0.0l],but between-group difference was not significant at 24 hours postinjury.Lower levels in MMP-9 were also observed in PJ34-treated group compared with injured group at 6 and 24 hours postinjury(P ＜ 0.05 or 0.01).Conclusion PARP inhibitor PJ34 can attenuate MMP-9 up-regulation,inhibit BBB injury and hence protect the brain against TBI in mice.

Objective To study the effect of siRNA targeting survivin and tumor necrosis factor related apoptosis-inducing ligand on the proliferation and apoptosis of hepatocellular carcinoma ( HCC) cells. Methods siRNA eukaryotic expression vector was constructed and the effects of soluble tumor necrosis factor related apoptosis-inducing ligand ( sTRAIL) on HCC cells were observed. Results The recombinant plasmid Psilence ( + )-survivin was successfully constructed. Survivin mRNA expression inhibition ratio reached 73% by RT-PCR. Observed by MTT method sTRAIL failed to inhibit the survival rate of HepG2、HepG2/Silence( - ) cells at 12 h、24 h、48 h when compared to control groups. With the survivin gene being inhibited, the survival rate of HepG2/Silence( + ) cells(0. 518?0. 017) decreased in 12 h compared to control groups (0. 741?0. 005 ) and reached the lowest level in 48 h ( P

Objective To explore the effect and clinical significance of expression of nuclear factor-?B((NF-?B)),ICAM-1 and COX-2 on the occurrence and metastasis of gastric carcinoma.Methods The(expression) of NF-?B,ICAM-1 and COX-2 in 142 patients with gastric carcinoma was examined by(immunohistochemical) SP technique.The adjacent gastric tissue(30 cases) served as a control group.Results The expression of NF-?B was 62.0% in gastric carcinoma tissue,much higher than that of the control group(P

AIM: To study the effects of hydrogen peroxied (H 2O 2) on cell proliferation and transcription of gelatinase A (MMP-2) and its inhibitor (TIMP-2) in vascular smooth muscle cells (VSMC). METHODS: Cell proliferation and toxicity by H 2O 2 were tested through MTT. The expression of MMP-2 mRNA and TIMP-2 mRNA in VSMC were evaluated by RT-PCR. RESULTS: The present study showed that H 2O 2 (more than 300 ?mol/L)was lethal to VSMC. 0 01-50 ?mol/L H 2O 2 promoted proliferation of VSMC in a time-dependant manner. A value (optical density) was reached to peak at 24 h after continuing stimulation of 10 ?mol/L H 2O 2. MMP-2/?-actin mRNA ratio significantly increased after stimulation with 1 ?mol/L?10 ?mol/L H 2O 2. TIMP-2/?-actin mRNA ratio was not significantly fluctuated at 12 h?24 h?36 h?48 h after continuing stimulation with 1 ?mol/L, 10 ?mol/L, and 50 ?mol/L H 2O 2.CONCLUSION: H 2O 2 at suitable concentrations stimulated proliferation of VSMC and induced transcription of MMP-2 gene in VSMC. There was no effect of H 2O 2 on transcription of TIMP-2 gene in VSMC. These results imply that H 2O 2 takes part in the pathological course of vascular remodeling through VSMC.

Objective To investigate the distribution characteristics of pathogens isolated fromcerebrospinal fluid of neurosurgical patients with intracranial infection following open craniotomy and thetherapeutic effect influenced by these pathogens,in order to give a reference to the clinical treatmentmeasures.Methods A retrospective analysis was made on the pathogen distribution and therapeuticeffect of 43 patients with intracranial infection and positive cerebrospinal fluid cultures after open cranioto-my from May 2007 to May 2013.Cerebrospinal fluid was cleared using the intraventricular catheter orlumbar catheter combined with intraventricular (ventricular irrigation) or intraspinal (intrathecalirrigation) injection of antibacterial agents.Results To test bacteria in cerebrospinal fluid pathogencultures,34 cases were infected with single strain (26 Gram-positive bacteria and 8 Gram-negativebacteria) and 9 cases had mixed infection with multiple strains.Fifty-two pathogen strains were isolated,including 32 (62％) Gram-positive bacteria,18 (35％) Gram-negative bacteria,2 (4％) fungi.A totalof 29 cases were cured (67％),7 improved (16％),and 7 ineffective (16％).Conclusions Cere-brospinal fluid pathogen infection is primarily Gram-positive bacterial infection,usually staphylococcusepidermidis and staphylococcus aureus.Gram-negative pathogens are acinetobacter,klebsiella,andpseudomonas aeruginosa.Ventriculoperitoneal shunting surgery and craniocerebral surgery are often asso-ciated with mixed infection of pathogens.Ventricular irrigation allows better results than intrathecal irriga-tion.Indications of intrathecal irrigation treatment used to control intracranial infection after ventriculoper-itoneal shunting surgery and craniocerebral surgery should be strictly performed.

Objective To explore the value of fluid-attenuated inversion recovery (FLAIR)sequence and gradient echo T2 ?weighted image (GRE-T2 ? WI)in diagnosis of acute traumatic extra-axial hemorrhage.Methods 50 patients who were diagnosed as acute traumatic extra-axial hemorrhage by plain CT underwent FLAIR and GRE-T2 ? WI in acute stage.The diagnosis consistency (Kappa ),detection rate of subarachnoid hemorrhage(SAH),epidural hemorrhage(EDH)and subdural hemorrhage(SDH)were compared.The detection rates of SAH in 8 locations among FLAIR,GRE-T2 ? WI and combination of two sequences were analyzed by Chi-square test.Results 48 patients were enrolled in the study.The diagnosis consistency of SAH between FLAIR and GRE-T2 ? WI was high (k =1.0).The detection rate of SAH was 100% for both FLAIR and GRE-T2 ? WI.Comparing with GRE-T2 ? WI and combi-nation of two sequences,FLAIR tended to misdiagnose SAH in perimesencephalic cistern (P <0.05).The diagnosis consistency of EDH between FLAIR and GRE-T2 ? WI was high (k =1.0).3 patients with EDH were all detected by FLAIR and GRE-T2 ? WI. The diagnosis consistency of SDH between FLAIR and GRE-T2 ? WI was high (k =0.943).The detection rate of 12 patients with SDH was 100% in FLAIR,and 1 patient with SDH was missed by GRE-T2 ? WI.Conclusion The detection rate of acute traumatic extra-axial hemorrhage is high for both FLAIR and GRE-T2 ? WI.Combination of two sequences can improve the accuracy of acute traumatic extra-axial hemorrhage in clinic.

BACKGROUND: Deletion of glial cell-derived neurotrophic factor and endothelin receptor type B gene will induce abnormal development of enteric nervous system. Neural stem cell transplantation can repair nervous system from anatomy and function,and be considered as a vector of gene transfection.OBJECTIVE: To transfect recombinant adenovirus carrying glial cell-derived neurotrophic factor and endothelin receptor type B gene into mouse neural stem cells, and to observe expression of target gene.DESIGN: A cell-gene study.MATERIALS: New-born Kunming mice were provided by the Animal Center of Tongji Medical College, Huazhong University of Science and Technology, China. jetPEI reagent was purchased from PolyPlus Co, France. The pAdTrack-CMV-GE with green fluorescent protein (GFP) was gifted by Doctor Sun Nianfeng and Zhang Jinghui in our laboratory.METHODS: Neonatal mouse brain tissues were sterilely obtained to prepare monoplast suspension. Adenovirus expressing glial cell-derived neurotrophic factor and endothelin receptor type B gene with GFP was dissolved in NaCI to prepare JetPEI/DNA complex. Subcultured neural stem cells in DMEM/F12 were regulated to 5×10~8/L, and 400 μL cell suspension and 100 μL JetPEI/DNA complex were seeded on a 24-well plate at 37 ℃ in 5% CO_2 incubator. Neural stem cells were harvested at 24, 48 and 72 hours following transfection.MAIN OUTCOME MEASURES: The efficiency of transfection was detected using fluorescence microscope and flow cytometry.Target gene expression in neural stem cells was determined using RT-PCR.RESULTS: Bright green fluorescence of the transfected cells could be observed under fluorescence microscope after 24 hours of transfection. The positive rate of GFP was 15.36%, 24.67%, 25.73% at 24, 48 and 72 hours following transfection respectively.Neural stem cells expressed glial cell-derived neurotrophic factor and endothelin receptor type B gene at various time points.Strap brightness was low at 24 hours, and exogenous gene expression was great at 72 hours.CONCLUSION: The target genes were successfully transfected into neural stem cells by using jetPEI reagent. Moreover, glial cell-derived neurotrophic factor and endothelin receptor type B gene effectively transcribed and expressed in target cells.