Objective To investigate the effect of nalmefene on the cerebral ischemia-reperfusion (I/R) injury in rats.Methods Forty-eight male Sprague-Dawley rats, aged 3-4 months, weighing 220260 g, were randomly allocated to control group (group C), sham operation group (group S), cerebral I/R group (group I/R), or nalmefene group (group N) using a random number table, with 12 rats in each group.Cerebral I/R was induced by occlusion of bilateral common carotid arteries for 20 min followed by reperfusion.Group C received no treatment.Group S underwent 20 min exposure of bilateral common carotid arteries and then received suture.In group N, nalmefene 0.1 mg/kg was injected intraperitoneally immediately after reperfusion.At 6, 24 and 72 h of reperfusion, venous blood samples were collected for determination of the concentrations of S-100β protein and neuron-specific enolase (NSE) in plasma by enzyme-linked immunosorbent assay.After the last blood sampling, the rats were sacrificed, and brains were removed for determination of interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents in brain tissues by enzyme-linked immunosorbent assay.Results Compared with group C, the plasma S-100β protein and NSE concentrations at each time point of reperfusion, and TNF-α and IL-1βcontents in brain tissues were significantly increased in S and I/R groups (P＜0.01).Compared with group S, the plasma S-100β protein and NSE concentrations at each time point of reperfusion, and TNF-α and IL-1β contents in brain tissues were significantly increased in group I/R (P＜0.01).Compared with group I/R, the plasma S-100β protein and NSE concentrations at each time point of reperfusion, and TNF-α and IL-1β contents in brain tissues were significantly decreased in group N (P ＜ 0.01).Conclusion Nalmefene can mitigate cerebral I/R injury in rats.

0.05). ② The depth of anesthesia: the value of AEP index in AEP index group was significantly higher than that in control group ( P

Objective To investigate the clinical value of mean channel of neutrophil volume (MNV), mean channel of neutrophil conductivity (MNC) and mean channel of neutrophil scatter (MNS) in predicting acute bacterial infection.Methods Peripheral blood samples from 112 patients with positive blood cultures for bacteria,70 healthy subjects and 45 non-infectious subjects with high white blood cell count(WBC) were studied using the Coulter LH 750 hematology analyzer.MNV, MNC, MNS and neutrophil volume distribution width (NDW) were retrospectively analyzed and compared with total WBC, percentage of neutrophils,neutrophil left-shift and CRP.112 blood bacterial infections were grouped according to WBC count (A:WBC ＜11.0×109 /L;B:11.0×109/L≤WBC＜15.0×109 /L;C:WBC≥15.0×109 /L) and neutrophil rate (NE ＜ 0.85 and NE ≥ 0.85 ).Results MNV and NDW increased significantly in septic patients (154.17 ± 10.08,24.36 ± 4.14 ) compared with those of healthy control group (142.09 ± 4.13,19.04 ± 1.97) and non-infectious patients with high WBC group ( 150.63 ± 8.14,20.19 ± 4.73 ).There was statistically significant difference (F value were 20.738 and 28.190 respectively,P ＜ 0.01 ). On the contrary, MNS decreased significantly in septic patients (137.15 ± 7.61 ) compared with that of healthy group (144.51±4.36) and nonspetic patients with high WBC group (142.45±7.11) ,there was significant statistical difference (F=5.217,P＜0.01).The MNV, NDW and MNS of A group were 148.09±5.76,22.39±1.97,140.07±6.11 respectively.The MNV, NDW and MNS of B group were 152.83±5.75,24.14±1.35,141.44±5.35 respectively.The MNV, NDW and MNS of C group were 164.28±6.49,29.42±5.93,134.27±9.61 respectively. There was statistically significant difference compared with healthy group (F value were 24.720,31.642,7.931, P ＜ 0.01).The MNV, NDW and MNS in the group with NE ＜0.85 were 149.17±9.06,22.59±2.73,141.19±4.34 respectively.The MNV, NDW and MNS in the group with NE≥0.85 group were 159.03±10.23,27.64±4.51,135.62 ± 8.95 respectively.There was statistically significant difference compared with healthy group ( F value was 23.970,51.309,19.792,P＜0.01).With a cut-off of 150 for the MNV, a specificity of 90% and sensitivity of 70% were achieved.NDW was associated with neutrophil left-shift (r=0.33,P＜0.01).With a cut-off of 23 for the NDW, a specificity of 100% and a sensitivity of 72% were achieved.The sensitivity of the MNV and NDW was better than total white blood cells count (with a cut-off≥ 11.0×109 /L, the sensitivity was 57% ), percentage of neutrophils( with a cut-off≥0.85, the sensitivity was 44% ) and neutrophil shift to left ( with a cut-off ＞5%, the sensitivity was 66% ) and CRP (with a cut-off ≥10 mg/L, the sensitivity was 65% ).Conclusions The MNV and NDW of the neutrophil can reflect the morphologic change of neutrophil sensitively and specificialy in acute infection. As quantitative, objective and more sensitive parameters, MNV and NDW may have a potential role for predicting the acute bacterial infection.

Catharanthus roseus (L.) G. Don is a plant of the Catharanthus genus of Apocynaceae which has been reported to have therapeutic effects of detoxication and anticancer. In order to further study the alkaloid constituents of C. roseus, the aerial parts of the plant were extracted with 95% EtOH, and then treated with 2% H2SO4 and NH3H2O to obtain total alkaloids. The total alkaloids were separated and purified by column chromatography over silica gel and prepared by high performance liquid chromatography (HPLC). Their structures were elucidated on the basis of physicochemical properties and spectral data. A new alkaloid together with five known compounds were isolated and identified as vindolinine B (1), lochnericine (2), horhammericine (3), vindorosine (4), vindoline (5), and coronaridine (6). Compound 1 is a new compound and named as vindolinine B.

Objective To evaluate the effect of surgical trauma on the cognitive function and expression of hepcidin and ferroportin 1 (FP1) in hippocampus in aged rats.Methods One hundred male Sprague-Dawley rats,aged 18 months,weighing 400-500 g,were randomly divided into 2 groups with 50 rats in each group:control group (group C) and surgical trauma group (group ST).The rats were anesthetized with chloral hydrate,but underwent no operation in group C.The rats Were anesthetized with chloral hydrate and underwent 30 min of modified exploratory laparotomy in group ST.Ten rats were chosen from each group at 24 h after operation and the cognitive function was assessed using Morris water-maze test for 6 consecutive days.Ten rats were sacrificed on 1st,3rd,5th and 7th days after beginning of Morris water-maze test and brains were removed for determination of hepcidin and FP1 expression in hippocampus by PCR and Western blot.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the original platform quadrant and frequency of crossing the original platform were decreased on 3rd,4th and 5th days after beginning of Morris water-maze test,and the expression of hepcidin was up-regulated and the expression of FP1 was down-regulated at each time point in group ST (P ＜ 0.05).Conclusion Surgical trauma can decrease the cognitive function in aged rats and the mechanism may be related to up-regulation of hepcidin expression and down-regulation of FP1 expression in hippocampus.

Objective To study the methods and outcome of 71 patients with Parkinson's disease treated with microelectrode-guided stereotactic pallidotomy and thalamotomy. Method Pallidal and thalamal target sites are chosen by supervision of microelectrode recording technique in 71 patients with Parkinson's disease. The UPDRS motor score was used to evaluate the outcomes 12 weeks before and after operation Result After 12 months follow-up, tremor disappeared completely or nearly completely in 12 patients who underwent unilateral and l bilateral ventrolateral thalamotomy. Dramatic improvement of tremor, rigidity, bradykinesia were observed in 57 patients underwent posteroventral pallidotomy,including 6 underwent bilateral posteroventral pallidotomy. Intracerebral hemorrhage was observed in l patient. Conclusion Microelectrode-guided stereotactic pallidotomy and thalamotomy are effective in treatmenting Parkinson's disease, but with serious complications

AIM: To study the effects of tumor necrosis factor-α (TNF-α) on the transcriptional activity of nuclear factor-erythroid 2-related factor 2 (Nrf2) in pulmonary microvascular endothelial cells. METHODS: Rat pulmonary micro-vascular endothelial cells (PMVECs) were cultured by lung tissue block pasted methods, and identified immunocytochemically using Ⅷ factor-related antigen. The cells were treated with different doses TNF-α (prepared in serum-free medium) for 4 h. Subcellular localization and levels of Nrf2 in PMVECs were observed with immunocytochemical methods. Nuclear extract were obtained to assayed transcriptional activity of Nrf2 with EMSA. Total RNA were isolated to assay the mRNA expression of Nrf2 by RT-PCR. RESULTS: The protein level of Nrf2 in the nuclei and transcriptional activity increased dose-dependently in PMVECs after treated with TNF-α at concentrations of 2.5, 5.0 or 10.0 μg/L. However, the protein level of Nrf2 in nuclei and transcriptional activity decreased dose-dependently in PMVECs after treated with TNF-α at concentrations of 20 or 40 μg/L. No different mRNA expression of Nrf2 in PMVECs treated with TNF-α at all concentration above was observed. CONCLUSION: Transcriptional activity of Nrf2 increases in PMVECs treated with low or moderate doses of TNF-α and decreases in PMVECs treated with high doses of TNF-α.

Objective To study the role of human carcinoembryonic antigen-related cellular adhesion molecule 1 (hCEACAM1)in mediating the specific adhesion of N. gonorrhoeae to its human host cells.Methods A recombinant eukaryotic expression vector pCDPGICEA1 was constructed by putting hCEACAM1 cDNA behind both hCD46 promoter and rabbit β-globulin intron 2,and with which,the COS-1 cells were transfected. Following G418 selection, the COS-1 cells expressing hCEACAM1 were sorted out with flow cytometry. The adhesion of N. gonorrhoeae to the gene transfected COS-1 cells was analyzed with bacterial binding assay. Results hCEACAM1 cDNA could be expressed effectively under the direction of hCD46 promoter and rabbit β-globulin intron 2,and N. gonorrhoeae could adhere to COS-1 cells expressing hCEACAM1. Conclusion hCEACAM1 can mediate the adhesion of N. gonorrhoeae to animal originated COS-1 cells. thus its transgenic mice may be used as a novel animal model for studying N. gonorrhoeae infection.

Objective To observe the effect of Midazolam on caspase-3 activation of cortical neurons in rat brain Methods The experiment consisted of part A and part B,including 64 7-day-old Sprague Dawley rats of either sex In part A 24 rats were randomly divided into 4 groups according to the time point when the arterial blood was collected after intraperitoneal injection of 180 mg/kg Midazolam(n=6 in each group),group A1 without Midazolam treatment as control,group A2 (15 min),group A3 (30 min),group A4 (60 min) In part B,40 rats were randomly divided into groups B2-B5 (n=8 in each group) according to the dose of Midazolam (45,90,145,180 mg/kg) and group B1 as control receiving 180 mg/kg normal saline Both control and experimental rats were perfused transcardially with paraformaldehyde 6 h after Midazolam treatment,then their brains takenout were used for caspase-3 examination by immunohistochemistry Results Single injection of Midazolam did not induce hypoxia/ischemia in 7-day-old SD rats,but led to the activation of caspase-3 in cerebral neurons Furthermore,higher single dose of Midazolam enhanced the activation of caspase-3 more Conclusion Midazolam triggers the activation of caspase-3 in cortical neurons and may lead to neuronal apoptosis

OBJECTIVE: To investigate the effects of human cerebrospinal fluid (CSF) during development phase on migration and differentiation of fetal brain neural stem cells (NSCs).METHODS: Fetal brain cells of gestational age of 16 weeks that were frozen in liquid nitrogen were obtained, resuscitated and incubated in DMEM/F12 medium containing epithium growth factor (EGF), basic fibroblast growth factor (bFGF), B27 and N2. The neurospheres cultured for 14 days were obtained. CSF was absorbed from the subarachnoid cavity and brain ventricle in the embryonic group. CSF was collected by lumbar puncture or ventricular puncture in the child group. The neurospheres cultured for 14 days were transplanted into the pure CSF in an incubator containing 5% CO_2 at 37 ℃. Cellular migration and growth of neurospheres in CSF were observed. Effects of CSF on neural cell differentiation were identified by immunofluorescence. RESULTS: Neural stem cells in the form of neurospheres derived from fetal brain were inoculated into the pure CSF, and cell migration were commonly observed besides few of neurospheres in child CSF culture at 6 hours following culture. Surrounding cells of neurospheres extended processes, forming cell cord that became cell webs after extension. Compared with the embryonic group, positive rate of glial fibrillary acidic protein was significantly increased in the children group (P < 0.01), but positive rates of nerve fiber and nestin were significantly decreased (P < 0.01). In addition, galactocerebroside-positive cells were only found in 3 baby CSF cultures. CONCLUSION: There existed significant affections on both migration and differentiation of human neural stem cells when cultured in pure CSF with different developmental phase, suggesting that CSF is one of major niche factors for central neural system development.