Objective To explore the effects of coriafia lactone (CL)-activated astrocytes (Ast) conditioned medium (ACM) on the expressions of glutamate (Glu) and GluR2 in the brain of rat. Methods Asts of hippocampus were cultured according to the McCarthy and DeVellis's method, and then the ACM was collected. Forty-eight male adult Sprague-Dawley (SD) rats were randomly divided into the control group (n=16) and the CL group (n=32). Rats in the control group were administered 10 μL ACM I. C. V., which was not added any stimulating substance. Rats of the CL group were injected I. C. V. 10 μL CL-activated ACM. The rats in both groups were subdivided into post-injection 2,4,8,12h subgroups, 4 in each subgroup in the control group and 8 in each subgroup in the CL group. The behaviors of the rats were observed and the expressions of Glu and GluR2 in the cerebral cortex and hippocampus were detected with immunohistochemistry and immunofluorescence. The content of GluR2 was tested with Western blot. Results The rats injected with CL-activated ACM showed seizure activities, whereas the rats of the control group showed no seizure activities. The expression of Glu in cerebral cortex and hippocampus in the brains injected with CL-activated ACM was increased compared with the control group at 4h (P<0.05), but the expression of GluR2 was attenuated compared with the control group at 4h(P<0.05). The results of GluR2 in the cerebral cortex and hippocampus detected with Western blot were different significantly with control group (P<0.05). Conclusion CL-activated ACM can enhance the expression of Glu and reduce the expression of GluR2 in the brain of rat, resulting in the activation of AMPA pathway and the Ca2+ influx, and then induce seizure activities.

Objecfive To study the effect of Danhong injection(DI)on intimal hyperplasia and expressions of basic fibroblast growth factor(bFGF)and tissue factor(TF)in carotid arteries of rabbits after percutaneous transluminal angioplasty(PTA). Methods Fifty New Zealand White rabbits were randomly divided into 5 groups (n=10),namely sham-operated group,model group,and DI treatment groups(low-,medium-and high-dose DI treatment groups).The animals in the sham-operated group were only given external carotid artery ligation.The rabbit models of carotid stenosis in the later 4 groups were established by ballon-injury.Before the carotid balloon injury,the animals in the DI treatment groups were given DI at doses of 1 mL/kg·d-1,2mL/kg·d-1 and 4 mL/kg·d-1,respectively,for 7 or 14 d via intravenous injection;the animals in the sham-operated and model groups were given saline solution(2mL/kg·d-1)instead.The animals were sacrificed and their common carotid arteries were removed on the 7th and 28th d of surgery;morphology changes of the arterial wall with hematoxylin eosin staining were observed by optical microscope,and the expressions ofbFGF and TF of the vascular wall were detected through immunohistochemical techniques;with the help of an image analysis system,semiquantitative analysis was performed.Results On the 7th d of surgery,the intimal area and intima area/media area (I/M)ratio in the model group showed no significant differences as compared with those in the other 4groups(P>0.05).On the 28th d of surgery,the intimai area and I/M ratio in the model group were increased markedly as compared with those in the sham-operated group (P<0.05), and the intimal area and I/M ratio in the medium and high dose DI treatment groups were significantly reduced as compared with those in the in the model group (P<0.05). The expressions of TF and bFGF in the neointima of carotid arteries of rabbits treated with medium and high dose of DI were significantly weaker than those of the model group (P<0.05), and those of rabbits treated with low dose of DI had no obvious differences as compared with those of the model group (P>0.05). Conclusion Medium and high dose of DI can inhibit the intimal hyperplasia resulting from percutaneous transluminal angioplasty, might through the inhibition of expressions of bFGF and TF in the neointima of the arterial wall.

BACKGROUNDIt is known that excessive release of glutamate can induce excitotoxicity in neurons and lead to seizure. Dexamethasone has anti-seizure function. The aim of this study was to investigate glutamate-dexamethasone interaction in the pathogenesis of epilepsy, identify differentially expressed genes in the hippocampus of glutamate-induced epileptic rats by mRNA differential display, and observe the effects of dexamethasone on these genes expression.

METHODSSeizure models were established by injecting 5 microl (250 microg/microl) monosodium glutamate (MSG) into the lateral cerebral ventricle in rats. Dexamethasone (5 mg/kg) was injected intraperitoneally at 30 minutes after MSG inducing convulsion. The rats' behavior and electroencephalogram (EEG) were then recorded for 1 hour. The effects of dexamethasone on gene expression were observed in MSG-induced epileptic rats at 1 hour and 6 hours after the onset of seizure by mRNA differential display. The differentially expressed genes were confirmed by Dot blot.

RESULTSEEG and behaviors showed that MSG did induce seizure, and dexamethasone could clearly alleviate the symptom. mRNA differential display showed that MSG increased the expression of some genes in epileptic rats and dexamethasone could downregulate their expression. From more than 10 differentially expressed cDNA fragments, we identified a 226 bp cDNA fragment that was expressed higher in the hippocampus of epileptic rats than that in the control group. Its expression was reduced after the administration of dexamethasone. Sequence analysis and protein alignment showed that the predicted amino acid sequence of this cDNA fragment kept 43% identity to agmatinase, a member of the ureohydrolase superfamily.

CONCLUSIONSThe results of the current study suggest that the product of the 226 bp cDNA has a function similar to agmatinase. Dexamethasone might relax alleviate seizure by inhibiting expression of the gene.

Animals ; Base Sequence ; Dexamethasone ; pharmacology ; Electroencephalography ; drug effects ; Epilepsy ; chemically induced ; drug therapy ; genetics ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Male ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Sodium Glutamate ; pharmacology

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.
Animals ; Autophagy ; Base Sequence ; DNA Primers ; Hypoxia ; physiopathology ; In Vitro Techniques ; Lung ; physiopathology ; Lysosomes ; physiology ; Oxygen Inhalation Therapy ; Rats ; Real-Time Polymerase Chain Reaction

OBJECTIVETo observe the effects of repeated + Gz exposures on the apoptosis of myocyte in rats.

METHODSTwelve male Sprague-Dawley rats were randomly divided into three groups: Control group, + 6Gz group and + 10Gz group. The rats of + Gz groups were exposed to + 6Gz for 3 min, + 10Gz for 3 min respectively, 1 b/d, 1 week. Four control rats were kept at the Earth gravity (1G) in the room with the centrifuge. All animals were anaesthetized and anatomies 1 day after the last exposure. Ventricular myocardium was studied by electron microscopy and terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) method.

RESULTSThe apoptosis of myocyte in control group and + 6Gz group were scarcely observed by electron microscopy, while heterochromatin concentration and margination were observed in + 10Gz group. The apoptotic index of myocardium increased significantly in + 6Gz and + 10Gz group compared with that of the control group (P < 0.05) and showed the largest value in the + 10Gz group (P < 0.05).

CONCLUSIONRepeated + Gz exposures may induce apoptosis in myocyte, and the number of apoptosis in myocyte increases gradually with the increase of G value.

Acceleration ; Animals ; Apoptosis ; physiology ; Male ; Myocardium ; cytology ; Myocytes, Cardiac ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo make multi-central clinical evaluation of the massage for supplementing qi and removing obstruction in the Governor Vessel for treatment of infantile diarrhea due to spleen deficiency.

METHODSBy using multi-central, randomized and controlled method, 275 cases were randomly divided into an observation group (n = 137) and a control group (n = 138). The observation group were treated by the massage for supplementing qi and removing obstruction in the Governor Vessel, and the control group by routine massage therapy in Tuina Science, a teaching material for college and school of TCM. After treatment for 7 days, their therapeutic effects were compared.

RESULTSThe cured rate was 83.2% in the observation group and 69.6% in the control group with a signifi cant difference between the two groups (P < 0.05), the former being better than the latter. The mean cured time was (3.22 +/- 1.04) days in the observation group and (4.20 +/- 1.11) days in the control group with a significant difference between the two groups (P < 0.05), the former being shorter than the latter.

CONCLUSIONThe massage for supplementing qi and removing obstruction in the Governor Vessel has a definite therapeutic effect on infantile diarrhea due to spleen deficiency, with rapid effect.

Combined Modality Therapy ; Diarrhea, Infantile ; therapy ; Female ; Humans ; Infant ; Male ; Massage ; Qi ; Splenic Diseases ; therapy

AIMTo study the clinical effects of a disposable circumcision device in treatment of male patients of different ages with either phimosis or excess foreskin.

METHODSOne thousand two hundred patients between the age of 5 and 95 years underwent circumcision using this procedure in the 2-year period between October 2005 and September 2007. Of these cases, 904 had excess foreskin and 296 were cases of phimosis.

RESULTSIn 96.33% of the cases the incision healed, leaving a minimal amount of the inner foreskin with no scarring and producing good cosmetic results. There were no incidents of device dislocation or damage to the frenulum. The average operative time was 2.5 min for excess foreskin, and 3.5 min for phimosis. During the 7 days of wearing the device, mild to moderate edema occurred in 10.08% of cases with excess foreskin and in 2.58% of those with phimosis. Edema in the frenulum was seen in 1.67% of patients, and only 0.67% had an infection of the incision. A total of 86.25% of patients reported pain due to penile erection. After removal of the device, 0.58% of the cases had minimal bleeding around the incision, and 2.42% had wound dehiscence.

CONCLUSIONThe new device can be applied to an overwhelming majority of patients with phimosis and excess foreskin. This technique is relatively simple to perform, and patients who underwent this surgery had very few complications. Antibiotics were not required and patients reported less pain than those who were circumcised using conventional methods. Circumcision with this device requires minimal tissue manipulation, and is quicker and safer than circumcision using conventional techniques.

Adolescent ; Child ; Child, Preschool ; Circumcision, Male ; methods ; Humans ; Male ; Minimally Invasive Surgical Procedures ; Pain, Postoperative

OBJECTIVETo determine the relative resistance to HIV-1 infection of CD4 + T lymphocytes in HIV-exposed seronegative individuals (ESNs) in China.

METHODSHIV primary isolates were obtained from peripheral whole blood of HIV-infected persons. CD4 + T lymphocytes of Chinese ESNs were separated from peripheral blood mononuclear cells with magnetic cell sorting (MACS). The purified CD4 + T lymphocytes were cocultured with HIV primary isolates. The p24 level was detected and the culture medium was refreshed every 3 days within 2 weeks.

RESULTSFor M tropic HIV strains, p24 level was significantly lower in ESN group than in control group (P < 0.05); for some M tropic HIV strains, even no p24 replicated in ESN group. However, T tropic virus strains had no significant difference between these two groups (P > 0.05).

CONCLUSIONCD4 + T lymphocytes of Chinese ESNs may possess relative resistance to M tropic HIV strains, which may be one of the main influencing factors that result in ESN.

Adult ; CD4-Positive T-Lymphocytes ; immunology ; virology ; China ; Female ; HIV ; classification ; isolation & purification ; pathogenicity ; HIV Infections ; virology ; HIV Seronegativity ; immunology ; Humans ; In Vitro Techniques ; Male ; Sexual Partners

OBJECTIVETo investigate the effects on ribbon synapse of inner hair cells after superpulsed CO2 laser-assisted cochleostomy in SD rats.

METHODSEighteen SD rats were randomly divided into laser-assisted surgery groups (2 W group and 5 W group), sham-operated group and control group. Ten of those were performed a cochleostomy using superpulsed CO2 laser with a corresponding power. Auditory brainstem responses (ABR) were measured pre-and postoperatively. The ribbon synapses at apical and middle cochlear turns were observed under laser scanning confocal microscope and then were quantified with 3ds Max software.

RESULTSThe postoperative ABR thresholds of the 2 W and 5 W groups were larger than the preoperative case (t = -5.65, P < 0.01; t = -4.97, P < 0.01). The synapse number at the middle turn decreased significantly in 5 W group (F = 17.15, P < 0.01), while no significant changes were noted at the apical turn (P > 0.05). There was no statistical difference in 2 W group (P > 0.05).

CONCLUSIONSThe superpulsed CO2 laser-assisted cochleostomy with high-power is accompanied by a synaptic injury, while no obvious effects after the low-power laser surgery, which might be a safe strategy to preform cochleostomy.

Animals ; Cochlea ; surgery ; Hair Cells, Auditory, Inner ; radiation effects ; Laser Therapy ; Lasers, Gas ; adverse effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Synapses ; radiation effects

The present study was undertaken to investigate the role of glial glutamate transporter-1 (GLT-1) in the brain ischemic tolerance induced by cerebral ischemic preconditioning (CIP) by observing the effect of GLT-1 antisense oligodeoxynucleotides (AS-ODNs) on the neuro-protection of CIP against brain ischemic insult in rats. Wistar rats with permanently occluded bilateral vertebral arteries were randomly assigned to 7 groups: (1) Sham group: the bilateral common carotid arteries (BCCA) were separated, but without occluding the blood flow; (2) CIP group: the BCCA were clamped for 3 min; (3) Brain ischemic insult group: the BCCA were clamped for 8 min; (4) CIP+brain ischemic insult group: 3 min CIP was preformed 2 d prior to 8 min ischemic insult; (5) Double distilled water group: 5 muL double distilled water was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively; (6) AS-ODNs group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after the BCCA was separated (but without occluding the blood flow), respectively. This group was further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs; (7) AS-ODNs+CIP+brain ischemic insult group: 5 microL AS-ODNs solution was injected into the right lateral cerebral ventricle 12 h before, 12 h and 36 h after CIP, respectively. This group was also further divided into 9 nmol and 18 nmol subgroups according to the doses of AS-ODNs. The other treatments were the same as those in CIP+brain ischemic insult group. The effect of the AS-ODNs on the expression of GLT-1 was assayed by using Western blot analysis. The profile of delayed neuronal death (DND) of pyramidal neurons in the CA1 hippocampus was evaluated by using thionin staining under light microscope by determining the neuronal density (ND) and histological grade (HG). Western blot analysis showed that AS-ODNs injected into the lateral cerebroventricle inhibited the expression of GLT-1 in the CA1 hippocampus in a dose-dependent manner. Neuropathological evaluation showed that there was no apparent DND in sham and CIP groups. Obvious DND of pyramidal neurons was found in brain ischemic insult group, which was represented by an increase in HG and a decrease in ND. CIP effectively protected the pyramidal neurons in the CA1 hippocampus against DND normally induced by ischemic insult, which indicating that CIP induced ischemic tolerance on the pyramidal neurons in the CA1 hippocampus. However, the injection of AS-ODNs into the lateral cerebroventricle blocked the neuro-protection of CIP against DND induced by brain ischemic insult. These results further proved the role of GLT-1 in the brain ischemic tolerance induced by CIP in rats.
Animals ; Brain ; pathology ; Brain Ischemia ; drug therapy ; CA1 Region, Hippocampal ; pathology ; Excitatory Amino Acid Transporter 2 ; metabolism ; Ischemic Preconditioning ; Oligodeoxyribonucleotides ; pharmacology ; Oligonucleotides, Antisense ; pharmacology ; Pyramidal Cells ; metabolism ; Rats ; Rats, Wistar