Objective To investgate the role of aquaporin-4(AQP4) in secondary cerebral edema after ischemia/reperfusion injury in rats.Methods When the models of reversible middle cerebral artery occlusion were established, the alterations of cerebral edema and BBB were evaluated by measuring water and Eval's Blue (EB) contents of cerebral tissue, and the expression of AQP4 in brain was observed by Western Blot at different time point after reperfusion. At last, the correlation between expression of AQP4 and water and EB contents of cerebral tissue were analysed.Results There were found that water and EB contents of cerebral tissue in rat models significantly higher than those of control group at different time point after ischemia/reperfusion ( P

Objective To investigate the effect of U0126 on brain edema and the expression of aquaporin 4 (AQP4) in rat brains after cerebral ischemic injury.Methods Totally 48 healthy male SD rats were divided randomly into ischemia group, treatment group and normal group.Rats in ischemia group and treatment group underwent middle cerebral artery occlusion by using an intraluminal thread method.Thirty minutes before operation, the rats in treatment group were injected into lateral cerebral ventricle with U0126, while rats in ischemia group accepted normal saline.24 hours after operation, the water content and Evans Blue in rat brains were determined as to exploring the degree of brain edema.Immunohistochemistry,Western blot and RT-PCR technique were applied to detect AQP4, p-ERK1/2 and p-ELK1.Results Compared with normal group, the water content and AQP4 expression in ischemia group were increased obviously.The water content and AQP4 expression in treatment group (protein:149.0?1.1,mRNA:0.328?0.010) were lower than those in ischemia group (protein:153.6?0.8,mRNA:0.400?0.015,P

Objective To study the influence of nitric oxide synthase (NOS) inhibitor in a rat model of Parkinson’s disease. Methods Hemi-Parkinsonism rat model was established by stereotaxic 6-hydroxydopamine (6-OHDA) lesions in striatum in which nitric oxide synthase (NOS) was inhibited by L-Nitro-Arginine (L-NNA) and apomorphin-induced rotational behavior was measured. The immunohistochemical staining method was used to observe the change of striatal nNOS-positive neurons and nigral tyrosine hydroxylase(TH)-positive neurons. Results L-NNA dramatically protected 6-OHDA-injected rats against indices of severe injury to the nigrostriatal dopaminergic pathway, including decreases in numbers of TH-positive nigral neurons and rotational behavior. The nNOS-positive neurons showed no changes in numbers. Conclusions These results indicate that NO might mediate, in part, 6-OHDA-induced neurotoxicity and nNOS-positive neurons might resist the 6-OHDA neurotoxicity. NOS inhibitor may play a role in the protection of 6-OHDA neurons.

ObjectiveTo check the isolated heart by coronary angiography to discover the location, na-ture and degree of the coronary artery lesions more accurately and increase the comprehensive evaluation ability of cardiovascular disease.MethodsTen fresh isolated hearts with different causes of death were extracted and injected with barium sulphate as contrast substance by ring injector, then developed under Xper FD20 angiography equipment. The obtained pictures and image data were handled by three-dimen-sional angiography images with the software attached to the angiography equipment. The coronary artery tissues were HE stained and observed by microscope. The HE staining results were compared with the angiographic results.ResultsThe imaging data obtained from the 10 cases for examination showed 8 cases without coronary artery stenosis and 2 cases with Ⅲ, Ⅳcoronary artery stenosis, which were consistent with HE staining results of coronary artery organization and the both results were confirmed. ConclusionIsolated coronary angiography has an unique advantage for accurate grading of classification of coronary artery stenosis, examination of vascular malformation and tiny lesions, which can provide reference for the localization of small lesions and basis during the autopsy for identification conclusion.

Objective To detect the expression of human telomerase reverse transcriptase (hTERT ) and telomerase in hu‐man fetal gastric fibroblasts .Methods Human fetal gastric fibroblasts were isolated and cultured ,epithelial cells were exduded by CK‐18 immune staining .hTERT protein was determined by indirect immunofluorescence .Telomerase activity was analyzed using telomeric repeat amplification protocol assay (TRAP)while the same analysis in adult gastric fibroblast ,which as to be positive con‐trol .Results Cultured fibroblasts were CK‐18 negative .The positive hTERT immunostaining was detected in both cellular cyto‐plasm and nuclear compartments .Amplified telomeric repeats (about 170 bp) in human fetal gastric fibroblasts were longer than those (160 bp) in adult gastric fibroblasts .Conclusion hTERT and telomerase were expressed in human fetal gastric fibroblasts .

BACKGROUND: It is difficult to culture human dental pulp cells in vitro. Studies regarding effects of growth factors on proliferation and differentiation of dental pulp cells cultured in vitro have been reported. However, little is known about the Chinese herb rhizoma drynariae decoction on dental pulp cells cultured in vitro.OBJECTIVE: To observe the effects of different concentrations of rhizoma drynariae decoction on the proliferation and differentiation of human dental pulp cells cultured in vitro.DESIGN, TIME AND SETTING: A controlled observation was performed at the Scientific Resaarch Center, Fourth Hospital, Hebei Medical University between March 2006 and May 2007.MATERIALS: Human dental pulp cells were sourced from the patients who acquired orthotherapy through pulling out impacted wisdom tooth at the Department of Stomatology, Fourth Hospital, Hebei Medical University. Written informed content of sample collection was obtained from all patients. Rhizoma drynariae (place of production: Yunnan Province in China) was provided by the Dispensary of Traditional Chinese Medicine, Fourth Hospital, Hebei Medical University.METHODS: Human dental pulp cells were cultured in vitro using method of tissue piece. The effective ingredients of rhizoma drynariae were extracted by alcohol deposition. 1 mL of physic liquor contained 1 g crude drug and diluted into 10, 50, 100, 500, and 1000 mg/L culture medium utilizing fetal bovine serum. Subsequently, the prepared culture medium was used to culture human dental pulp cells in vitro. Cells that were cultured using culture medium without rhizoma drynariae decoction were used as controls.MAIN OUTCOME MEASURES: ①Primary culture and source identification of human dental pulp cells. ②Effects of different concentrations of rhizoma drynariae decoction on proliferation of human dental pulp cells by methyl thiazolyl tetrazolium (MTT) assay. ③ Effects of different concentrations of rhizoma drynariae decoction on fibronectin expression in human dental pulp cells by immunohistochemistry. ④ Effects of rhizoma drynariae decoction on ultrastructure of human dental pulp cells utilizing scanning electron microscope and transmission electron microscope.RESULTS: Primarily cultured human dental pulp cells displayed polygon- and shuttle-shaped appearance. Different concentrations of rhizoma drynariae decoctions, in particular 100 mg/L, exhibited proliferation-promoting effects on proliferation of human dental pulp cells, and could induce dental pulp cell synthesis and secrete fibronectin. Electron microscopy results revealed that following treatment of rhizoma drynariae decoctions, human dental pulp cells were found with abundant ridges on their surface, surround by extracellular matrix, cytoplasm full of abundant rough endoplasmic reticulum and dissociative ribosome, as well as evenly dispersed nuclear euchromatin, and occasionally seen heterochromatin.CONCLUSION: 100 mg/L rhizoma drynadae decoction apparently promotes the proliferation of human dental pulp cells cultured in vitro.

Objective To explore the role of inositol requiring enzyme 1 α (IRE1α) mediated endoplasmic reticulum stress associated apoptotic molecules in hippocampal neuronal injury in rats with status epilepsy following lithium-pilocarpine.Methods All 96 Wistar rats were randomly divided into control group and status epilepsy (SE) group.The SE group was further divided into 5 subgroups (3,6,12,24,48 h) according to different time points.pmmunofluorescence was used to observe the expressions of endoplasmic reticulum stress (ERS) markers glucose-regulating protein 78 kd (GRP78) and phosphoIRE1α (active form of endoplasmic reticulum resident protein IRE1α) at the CA3 area of rats in each group.Then,the expressions of IRElα mediated downstream apoptotie markers phospho-c-JunN-terminalkinase (JNK) and caspase12 were detected.Finally,TUNEL assay was used to observe neuronal apoptosis of hippocampal CA3 area at different time points after SE in rats.Results Immunofluorescence showed that GRP78 and phospho-IRE1α positive neurons were significantly increased in the SE subgroups compared with control group (6.90％ ± 0.96％,4.60％ ± 1.12％,respectively) and 12 h subgroup reached the peak (GRP78:87.45％ ±3.63％,F =356.82,P ＜0.05; phospho-IRE1α:86.90％ ±3.82％,F =300.80,P ＜ 0.05).Immunohistochemistry and Western blot demonstrated that the levels of phospho-JNK and caspase12 in the SE subgroups were significantly higher than that in the control group which reached the peak at 12 h after SE.The changes were in accord with phospho-IRE1α.Simultaneously,hippocampal neuronal apoptosis was detected in each SE subgroup and was most severe at 12 h after SE,which showed similar changes to the expressions of phospho-IRE1α,phospho-JNK and caspase12.Conclusions ERS was induced in rats following SE evidenced by increasing the expression of GRP78.IRE1α may promote hippocampal neuron apoptosis in rats following SE through activating JNK and caspase12.

Objective To investigate alterations of balance function in patients with mild-moderate Alzheimer's disease (AD) and with amnestic mild cognitive impairment (aMCI),and the possibility of using posturography to differentiate aMCI,mild-moderate AD and normal subjects. Methods The balance function of 20 patients with mild-moderate AD and 20 patients with aMCI were evaluated by posturography,and 20 healthy subjects of the same age range were recruited as controls.Results All posturography measures were significantly altered in mild-moderate AD patients compared with normal controls,with limits of stability( ( 15 398 ± 926 ) mm2 vs ( 31 654 ± 2132 ) mm2 ),open-eyed Mean X ( ( 10. 2 ± 4. 1 ) mm vs (5.8 ± 1. 4)mm) ,Mean Y(( -29.8 ± 10.2)mm vs ( -14.9 ±4.4) mm),Max X((30.5 ±9.5)mm vs (18.3 ±4. 1)mm ),Max Y((42.7 ± 11.4)mm vs (23.3 ±6.8)mm),LSKG((528.4 ± 105.4)mm vs (390. 3 ± 68.4 ) mm ),SSKG ( ( 252. 5 ± 89. 7 ) mm2 vs ( 178.8 ± 40. 9 ) mm2 ),close-eyed Mean X ((13. 1 ±4. 5) mm vs (7.9 ± 1.5)mm) ,Mean Y (( -58.2 ± 16. 9) mm vs ( -25.6 ±5.4) mm) ,Max X ((37.7±10.5)mm vs (24.7 ±7.3) mm ),Max Y ((78.5±18.7)mm vs (39.9 ±9.9) mm),LSKG ((816.6±171.3) mm vs (533.5 ±97.4) mm),SSKG((649.0 ± 129.7) mm2 vs (290.5 ±73.3) mm2),respectively ( t = 8.57; open-eyed F = 17.41,38. 10,60. 46,102. 10,29. 31,27. 85; close-eyed F = 37.20,541.79,34. 51,185.56,122. 83,384. 27 ;all P ＜0. 05) ;limits of stability ( (23 921 ± 1637 )mm2 vs (31 654 ±2132 ) mm2 ) and mean Y ( Antero-posterior sway,( - 39. 8 ± 8. 6 ) mm vs ( - 25.6±5.4 ) mm) were the only parameters which discriminated between aMCI and normal controls,respectively ( t = 6. 50,P = 0. 038; t =- 15.34,P = 0. 012). Conclusions Impairment in balance is a feature not only of mild-moderate AD,but also of aMCI,and posturography may be used as a possible test in differentiating between normal subjects,patients with aMCI and patients with mild-moderate AD whose motor performance and balance features are otherwise clinically normal,limits of stability and mean Y are the most sensitive parameters.

Objective To observe the morphological changes of rats' pancreas and nitric oxide (NO) and endothelin(ET) in the blood serum in rats after exposure to different pulses of high power pulse microwave (HPPMW).Methods SD-rats were irradiated with 104,105 and 4 × 105 pulses of HPPMW,respectively.After gloss observation,the histopathological changes of pancreas were observed through biological microscope and electroscope.The changes of amylase,nitric oxide and endothelin in blood serum were detected by biochemical and radio-immunological methods. Results Compared with the blank control,no apparent abnormality could be observed in the pancreas of all groups.The dilatation of capillary could be observed in each experimental group by microscope.The ultrastructure changes of pancreas were most serious in 4 × 105 pulse group,especially at 24 and 48 h after irradiation.Compared with the control group,the levels of serum amylase were decreased (F =12.58,11.73,P ＜ 0.05),while ET were increased (F =4.50,4.49,P ＜0.05) at 24 and 48 h after irradiation.The levels of NO in serum were increased ( F =17.51,41.72,19.98,32.64,P ＜ 0.05 ) at each time-point.The level of NO went up with the increase of pulses.Conclusions HPPMW has damage effects on the pancreas in rats.The pulses with the pancreas can lead to severity of the damage. The mechanism of HPPMW may be involved in the enhancement of ET and NO in serum.

Objective To investigate the cytotoxicity of silica nanoparticles on vascular endothelial cells, and to clarify its action mechanism.Methods The 60 nm silica nanoparticle was selected and the invitro cultured human umbilical vein endothelial cells (HUVECs)were used as cell model.The HUVECs were divided into control and silica nanoparticle exposure groups with concentrations of 12.5,25.0,and 100.00 mg·L-1 .MTT assay was used for the determination of cell viability,lactate dehydrogenase (LDH)release assay for membrane integrity,flow cytometry (FCM)for intracellular reactive oxygen species (ROS)content,and real-time PCR assay for intracellular NF-E2-related factor 2 (Nrf2 ), heme oxygenase-1 (HO-1 ), superoxide dismutase 2 (SOD2 ) and glutamate-cysteine ligase catalytic subunit (GCLC)mRNA levels.Results The MTT results showed that the cell viabilities in each silica nnaoparticle exposure group were decreased compared with control group in a dose-dependent manner. Upon the silica nanoparticle exposure for 12 h,the cell viability was declined significantly only in 100 mg·L-1 exposure group compared with control group (P<0.05).When exposured for 24 h,the cell viabilities in 25.0, 50.0,and 100.0 mg·L-1 exposure groups were declined significantly compared with control group (P<0.05). Under the exposure to silica nanoparticle with the same dose, the cell viabilities were decreased along with the elongation of exposure time.LDH assay and FCM showed that except for that in 12.5 mg·L-1 exposure group, both the LDH activities in media and intracellular ROS levels in other exposure groups were increased compared with control group (P<0.05 ). The results of real-time fluorescence PCR showed that the mRNA levels of Nrf2, HO-1,SOD2 and GCLC in 100 mg·L-1 silica nanoparticle exposure group were increased significantly compared with control group (P<0.05).Conclusion Silica nanoparticles have toxicity to vascular endothelial cells,which includes reducing cell viability,membrane integrity destruction,induction of ROS generation,and tranSCriptional regulation of redox-related factors. Oxidative damage is one of the mechanisms of vascular endothelial toxicity mediated by silica nanoparticles.