Objective To investigate the responses of astrocytes and neurons in rat lumbosacral spinal cord and medulla oblongata induced by chronic colonic inflammation,and the relationship between activated astrocytes and neurons.Methods Thirty-three male Sprague-Dawley rats were randomly divided into two groups.In treated group(n=17),colonic inflammation was induced by intraluminal administration of trinitrobenzenesulfonic acid(TNBS) in rats;In non-treated group(n=16),saline was intraluminally administered.The lumbosacral spinal cord and medulla oblongata were removed 3,7,14 and 28 days after intraluminal administration and processed for anti-GFAP,Fos and GFAP/Fos immunohistochemistry. Results Most activated GFAP positive astrocytes were distributed in the superficial laminae(Ⅰ-Ⅱ),intermediolateral nucleus(lamina Ⅴ),posterior commissural nucleus(laminae Ⅹ) and anterolateral nucleus(laminae Ⅸ) in lumbosacral spinal cord.Fos positive neurons were mainly expressed in the deeper laminae of the spinal cord(Ⅲ-Ⅳ,Ⅴ-Ⅵ).In the medulla oblongata,both GFAP-IR astrocytes and Fos-IR neurons were mainly distributed in medullary visceral zone(MVZ),which is composed of the nucleus of solitary tract(NTS),ventrolateral medulla(VLM) and intermediat reticular(IRt).The density of GFAP positive astrocytes in the spinal cord in treated rats 3,7 and 14 days after TNBS administration was significantly higher than that in non-treated rats(P0.05).Conclusions The astrocytes in lumbosacral spinal cord and medulla oblongata can be activated by colonic inflammation.The response of astrocytes decreased with the recovery of the colonic inflammation.Activated astrocytes are closely related to activated neurons in MVZ.

Objective The aim of this study was to investigate Fos expression in rat lumbarsacral spinal cord and medulla oblongata induced by chronic colonic inflammation. Methods Twenty six male Sprague Dawley rats were divided into three groups: group 1, colonic inflammation was induced in sixteen rats by intraluminal instillation of trinitrobenzenesulfonic acid (TNBS); group 2, saline was instilled intraluminally in eight rats; group 3, no stimulation was given in 2 rats. After 3, 7, 14 and 28 days of instillation, lumbarsacral spinal cord and medulla oblongata were removed and processed for Fos immunohistochemical staining. Results Fos neurons induced by TNBS instillation were mainly distributed in deep laminae (laminae Ⅲ Ⅳ,Ⅴ Ⅵ) in spinal dorsal horn and in medullary visceral zone in medulla oblongata. The number of Fos cells in the spinal cord and medullary visceral zone was significantly higher in rats after 7 and 14 day of TNBS instillation compared with that of controls. After 28 days of TNBS instillation, the number of Fos neurons in the medullary visceral zone decreased and became comparable to that of control group. However, the number of Fos cells (54.1?16.3) in the spinal cord in some rats was still significantly higher than that of controls (12.2 ?2.6, P

Objective To investigate the effects and potential mechanism of irinotecan (CPT-11), an antitumor drug, on human colorectal cancer cell line HT-29 and its impact on 4-amion pyridine (4-AP), a kalium ion channel blocker. Methods The effects of CPT-11, 4-AP and combination of two drugs on proliferation and invasion of HT-29 cells were measured by MTT and Transwell assay respectively. The impact of CPT-11 or 4-AP on cell apoptosis was determined by flow cytometry with Annexin-V and PI staining. The current of ATP sensitive potassium ion (IKATP) was measured by patch clamp. Results The CPT-11 could inhibit proliferation of HT-29 cells at dose from 1.0 to 64.0 μg/ml in dose-and time-dependent manners. Whereas the above effect was enhanced when CPT-11 combined with 4-AP (1.0 mmol/L). The administration of CPT-11 (16.0 μg/ml) or 4-AP (1.0 mmol/L) significantly induced the cell apoptosis and inhibited the invasion of HT-29 cells, furthermore, these effects could be enhanced by combination of two drugs. And the different concentrations of CPT-11 reduced the IKATP of cell membrane in negative dose-dependent manner. Conclusions The effects of CPT-11 on HT-29 cells, such as reducing proliferation and invasion as well as inducing the apoptosis, can be enhanced by 4-AP, which may be related to inhibition of ATP-sensitive potassium channels.

SUMMARY Here we report a case of cervical spondylosis misdiagnosed as cerebral infarction .The pa-tient was a 55-year-old man with a one-day history of weakness in his right extremities .Brain magnetic resonance imaging ( MRI) showed no acute abnormality , cerevical MRI showed that cervical spondylisis , C4/5 , C5/6 disc herniation , spinal canal stenosis and compression of the spinal cord .Then the patient was transferred to the Department of Orthopaedics and underwent surgical treatment of cervical spondylo -sis.Followed-up for six months , the weakness of his right extremities returned to normal .

Objective To study the effects of Irinotecan (CPT-11) on human colorectal cancer HCT-116 and HT-29 cells and investigate the potential mechanisms.Methods The effect of Irinotecan on the proliferation of HCT-116 and HT-29 cells was determined by MTT assays.The invasive capacity was measured by transwell assays,and the apoptosis of the tumor cells was detected by flow cytometry after stained with Annexin-V and PI.The difference between the current of ATP-sensitive potassium ion of HCT-116 and HT-29 was examined by patch clamp.Results It was found that 1.0-64.0 μg/ml CPT-11 could inhibit the proliferation and the invasive capacity of HCT-116 and HT-29 cells at both dose-and time-dependent manner.The IC_(50) of HCT-116 and HT-29 were 39.3 and 19.5 μg/ml respectively.Cytometry showed that the apoptotic rates were increased from 14.8% and 9.3% to 36.9% and 27.9% after the treatment of 32.0 μg/ml and 16.0 μg/ ml CPT-11,which were close to their IC_(50).The proportion of G_0/G_1 and S of HCT-116 and HT-29 was enhanced from 27.4% and 17.4% to 95.9% and 98.2%.Transwell assay indicated that the invasiveness of HCT-116 and HT-29 was reduced by 40.8% and 47.5%.The patch clamp showed that CPT-11 reduced the I_(KATP) of cell membrane at a negative dose-dependent manner.Conclusion CPT-11 could have a significant impact on the proliferation,invasiveness,cell cycle,and the apoptosis of human colorectal cancer cell HCT-116 and HT-29.HT-29 was more sensitive to CPT-11 than HCT-116.The inhibitory effect of CPT-11 on cell proliferation might be linked to its inhibition of ATPsensitive potassium channel.

ObjectiveTo determine the frequency of cerebral microembolism in patients with asymptomatic or symptomatic extracranial and intracranial arterial stenosis and to assess its relationship to the onset and course of ischemic stroke.MethodsTCD was used to monitor patients with extracranial and intracranial cerebral arterial stenosis. Double channel four-gated and power M-Mode were used to detect microembolic signals (MES). The recording time was 60 min and the number of MES was counted. Patients were divided into 2 groups as extracranial internal carotid artery (ICA) stenosis and middle cerebral artery (MCA) stenosis. Each group was divided into 3 subgroups as asymptomatic, acute ischemic stroke (<30 d) and old ischemic stroke (≥30 d).ResultsThe total number of monitored artery was 74 in 63 patients. In ICA stenosis with asymptom, old ischemic stroke and acute ischemic stroke, the frequency of MES was 0(0/10), 0(0/7), 33%(6/18) and the number of MES in acute ischemic stroke was 3, 9, 8, 10, 1, 40 (mean=11.8). In MCA stenosis with asymptom, old ischemic stroke and acute ischemic stroke, the frequency of MES was 14%(2/14), 14%(1/7), 39%(7/18) and the number of MES was 4 and 1 in asymptom, 1 in old ischemic stroke, 4, 1, 4, 15, 16, 1 and 29(mean=10) in acute ischemic stroke. In ICA stenosis, the difference between acute ischemic stroke and asymptom (P=0.013), between acute and old ischemic stroke (P=0.031) reached statistic significance.In MCA stenosis, there was no significant difference between acute ischemic stroke and asymptom (P=0.115), so as between acute and old ischemic stroke (P=0.214).ConclusionEmbolism was important in the mechanism of ischemic stroke due to extracranial and intracranial arterial stenosis. The closer to ischemic stroke onset, the higher the frequency and the number of MES. TCD monitor was helpful to study the pathogenesis of ischemic stroke due to extracranial and intracranial arterial stenosis and determine the treatment.

BACKGROUND: Electrochemically deposited coating, as an alternative method of plasma spraying coating, has arose widely attention, however, few reports concerning its in vivo biological features, especially the effect of coating on early osseointegration and duration of bone healing is poorly understood. OBJECTIVE: To investigate the early osseointegration of electrodepositied calcium phosphate and calcium phosphate/chitosan coatings on rabbits' femurs. DESIGN, TIME AND SETTING: An open experiment. The experiment was performed at the Key Laboratory for Oral Biomedical Engineering of Ministry of Education, Stomatology School of Wuhan University from April 2008 to May 2009. MATERIALS: Eighteen male Japanese rabbits were supplied by the animals' center of Wuhan University. Ti_6Al_4V alloy was purchased from Baoji Titanium Nickel Co., Ltd. Chitosan with over 75% degree of deacetylation was produced by American Sigma Company.METHODS: ① Cylindrical titanium alloy implants (3.3×8.0 mm) were designed with a gap (0.3×4.0 mm) in the middle part by using precision machine tool. The implants were prepared electrodeposited calcium phosphate coating surface (ELD group) and electrodeposited calcium phosphate/chitosan coating surface (ELDC group) with 2.5 mA/cm~2 electric current and 52 V temperature. Meantime, the implants with sandblasted surface were prepared as the control group. Totally 36 implants were randomly inserted into the distal femur condyles of 18 rabbits, and the new bone formation was labeled by fluorescence staining. MAIN OUTCOME MEASURES: At weeks 2 and 4 after operation, the implants were subjected to histological and histomorphometric analysis. RESULTS: All groups were observed different-quantity new bone formation. However, only ELDC group was seen fibrous tissues intervened at bone-to-implant interface. The ELD coatings were heterogeneously degraded, and the majority of the ELDC coatings were degraded. The new bone within gaps of ELD and control groups could be seen under the confocal laser scanning microscope with a continuous process of bone apposition. Percentages of bone-to-implant contact within and outside of the gaps of ELD group at weeks 2 and 4 were significantly higher than those of ELDC and control groups (P < 0.05). Bone formation rates of ELD group were significantly higher than those of ELDC group at weeks 2 and 4 after operation (P < 0.05). CONCLUSION: ELD implants can promote early osseointegration and induce bone tissue growth into the gaps; however, ELDC implants go against osseointegration of rabbit femurs.

Objective To investigate the association between the presence of cerebral microbleeds and chronic kidney disease in patients with ischemic stroke.Methods Patients with ischemic stroke within 1-6 months were consecutively recruited.Cranial MRI was taken within two weeks after recruitment.Cerebral microbleeds were assessed using Microbleed Anatomical Rating Scale on gradient echo MRI.Demographics including sex,age and risk factors were obtained.Chronic kidney disease was defined and classified according to National Kidney Fundation-Kidney Disease Outcome Quality Initiative Guideline.Glomerular filtration rate (GFR) was estimated by using the abbreviated Modification of Diet in Renal Disease equation.Results Of the 636 patients included,mean age was (59.8 ± 10.1) years,435 (68.4％) were male.Sixty-six had decreased estimated GFR (eGFR; ＜ 60 ml · min-1 · 1.73 m-2).Two hundred and one (31.6％) patients had cerebral microbleeds,which were most commonly located in deep or infratentorial location (133/201,66.2％).The presence of cerebral microbleeds was much higher in patients with decreased eGFR than the others (48.5％ (32/66) vs 29.6％ (169/570),x2 =9.709,P =0.002).Age,history of hypertension and decreased eGFR were associated with the presence of cerebral microbleeds in univariate analysis.In multivariate analysis,decreased eGFR was independently associated with the presence of cerebral microbleeds in deep or infratentorial location (OR =1.457,95％ CI 1.044-2.034,P =0.027),but not associated with the presence of cerebral microbleeds in pure lobe.Conclusion Impaired kidney function is associated with the presence of cerebral microbleeds in deep or infratentorial regions in patients with ischemic stroke.

Objective: To study the effect of docetaxet (DOC) combined with 4-AP on human breast can-cer MCF-7 cells and to explore whether 4-AP could strengthen the effect of docetaxel. Methods: MTT assays were performed to investigate the effect of docetaxel, 4-AP and the combination of them on the proliferation of MCF-7 cells. Flow cytometry was employed to detect cell cycles and cell apoptosis after the cells were stained by PI alone or by Annexin-V and PI. Results: Docetaxel could significantly inhibit the proliferation of MCF-7 cells in a dose- and time- dependent manner. 4-AP could inhibit the proliferation of MCF-7 cells and the inhibitory rates were 11.9%±1.7%, 42.1%±3.2%, and 44.2%±1.6% at 24h, 48h and 72h after adding 4-AP. Moreover 4-AP (5mmol/L) could strengthen the effect of docetaxel. 4-AP (25μmol/L) could increase the effect of Docetaxel. Docetaxel at 5μmol/L could significantly increase the percentage of cells at G_2/M (53.58%± 1.44% vs. 8.83%±0.44%, P<0.01) and decrease the percentage of cells at G_0/G_1 (11.48%±0.14% vs. 63.89%±0.98%, P<0.01), indicating that docetaxel blocked MCF-7 cells at G_2/M phase. 4-AP at 5mmol/L could in-crease the percentage of MCF-7 cells at G_0/G_1 and decrease the percentage of cells at G_2/M (0.42%±0.17% vs. 8.83%±0.44%, P<0.05). Docetaxel could significantly increase late apoptosis and death of MCF-7 cells af-ter treatment over 24h (from 6.97%±0.75% to 20.77%±0.75%, P<0.05). Docetaxel combined with 4-AP could increase early apoptosis rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05) and could increase late apopto-sis rate and death rate from 4.60%±0.91% to 12.20%±0.82% (P<0.05). Conclusion: Both docetaxel and 4-AP can inhibit the proliferation of MCF-7 cells. Docetaxel can increase the percentage of cells at G_2/M phase and 4-AP can increase the percentage of cells at G_0/G_1 phase. 4-AP could strengthen the inhibitory effect of docetaxel on the proliferation of MCF-7 cells through inducing cell apoptosis.

Objective To measure the activity energy expenditure(AEE) of healthy adults during level walking by using indirect calorimetry,and to analyze the characteristics and underlying influencing factors such as age and gender. Methods A total of 60 healthy adults aged 20-50 years (30 males and 30 femdes) participated in the study.All the subjects were divided into six groups by gender and age (the age span of each group was 10 years). The subjects were arranged to walk at speeds of 3.5,4.5,5.5km/h and run at 5.5,6.5,7.5km/h, respectively, on the treadmill. The resting energy expenditure ( REE ) and AEE were measured during walking and running at different speeds.There was a 5-minute rest among the test sessions. Results No difference in terms of AEE between the female and male at the same age ( P ＞ 0. 05 ). During 3.5km/h walking,AEE of 21-30 year-old females was lower than 31-40 yearold females and males( P ＜0.05 ) ;During 4.5km/h walking,AEE of 31-40 year-old females was higher than 21-30 yearold females and males and 41-50 year-old females; AEE of 21-30 year-old females was lower than 41-50 year-old males (P ＜ 0.05 ) ;During 5.5km/h walking,AEE of 31-40 year-old females was higher than 21-30 year-old females and males ( P ＜ 0. 05 ). During 6.5 km/h running, AEE of 31-40 year-old females was higher than 41-50 year-old females ( P ＜0.05), while during 7.5km/h running, AEE of 21-30 year-old males was higher than 41-50 year-old females (P ＜0.05 ). It was also found that the AEE of all groups except the 41-50 year-old females group was higher when walking at the speed of 5.5km/h than running at the same speed( P ＜ 0.05 ). Conclusions Age has more effect on REE and AEE than the gender. AEE of elder subjects is higher than that of the younger ones during walking, however, AEE of younger people increases faster than the elders during running. AEE of 31-40 year-old females is the highest in all groups both in walking and running. AEE in running is higher than in walking at the same speed.