Aim To develop an HPLC method for the determination of Aconitum alkaloids extracted from Radix aconiti preparata in rats. Methods Waters 2690@996 PAD system was used. The analytical column was a Halsil 100 C18 column (250 mm×4.6 mm ID, 5 μm). The mobile phase was water, methanol and diethyl amine at the ratio of 75∶ 25∶ 0.1. The flow rate was 0.9 mL·min -1. The wavelength of the detector was 240 nm. Results The linear ranges of aconitine in the heart, spleen, lung and kidney were 0.4-100 μg·mL -1, the correlation coefficients were 0.997 2, 0.998 6, 0.999 3 and 0.999 4, respectively. The linear range of aconitine in liver was 2-200 μg·mL -1 and the correlation coefficient was 0.999 0. The linear ranges of hypaconitine in heart, liver, spleen, lung, kidney, brain and spinal cord were 5-100 μg·mL -1, the correlation coefficients were 0.999 4, 0.999 7, 0.999 8, 0.998 4, 0.999 8, 0.999 8 and 0.999 7, respectively. Detection limits (S/N=3) of aconitine and hypaconitine were 0.4 μg·mL -1. The recoveries of aconitine and hypaconitine ranged from 88.7% to 102.2% and 86.5% to 101.3%, respectively, and the RSD of precision of aconitine and hypaconitine was 10%. Conclusion It appears to be an accurate and effective method that can offer reference basis for in toxication of Radix aconiti preparata clinically.

The cultivation of communication skill of anesthesiology medical students has already become a request as time developing.Not only it accommodates the request of medical model and society development to cultivate the communication skill of anesthesiology medical students from the aspect of strengthening their self-confidence and pride,global consciousness and international view,catholicity and humanity as well as cognitive ability,but also embodies the requirement of anesthesiology anthropo-culture.So there should be pertinent measures to cultivate the communication skill of anesthesiology medical students.

Objective To explore the clinical characteristics and risk factors of re-fracture in patients suffering from osteoporosis-related fractures as well as effective interventions.Methods From January 2006 to January 2008,a total of 273 patients with osteoporosis-related fracture were entered in the study,including out-patients and in-patients who were over 50 years old.The patients were divided into fracture group(n=225)and re-fracture group(n=48).The re-fracture rate was followed up for 2 years,during which 11 patients developed re-fracture.General data including age and sex,fracture types,femoral neck bone mineral density(BMD)T-scores tested by dual-energy X-rays absorptiometry(DEXA),Charlson index,timeinterval between two fractures as well as mobility skill assessment were collected and analyzed.Results The average age at the first fracture was 67.7±8.5 years vs.72.7±9.5 years for the re-fracture cases.Female accounted for 70.2% of the fracture group and 77.1% of the re-fracture group.The most common re-fracture type was vertebral fracture for the first time and femoral neck fracture for the second time during the followup.Risk factors for a second fracture in osteoporotic fractures patients include age(＞75 years,HR=1.23; ＞85years,HR=1.68),female sex(HR=1.36),prior vertebral fractures(HR=1.62),prior hip fractures(HR=1.27),BMD T-score＜-3.5(HR=1.38)and weakened motor skills(HR=1.27).The refracture rate in osteoporosis-related fractures was 4.9% followed up for 2 years.The second fracture happened 3.7 years after the first one on average.Conclusion The risks of second fracture among patients with initial brittle fracture are substantial.Mobility skill assessment is an important risk factor for osteoporosis fractures recurrence.There is adequate time between fracture and re-fracture for effective interventions to prevent or reduce the risks of refracture,especially for the old women with a vertebral or hip fracture.Medication,motor function rehabilitation and fall-down prevention training would be helpful.

ObjectiveTo observe and analyze the synapses developing process of newly generated connections of autologus activated Schwann cells (AASCs) in combination with fetal spinal cord cell suspension(FSCS) in the surrounding area of the spinal cord injury site.MethodsA total of 42 Wistar rats underwent unilateral ligation of the saphenous nerve.The portion of nerve tissues distal to the ligation site were harvested 1 week after operation.AASCs were isolated,cultured and purified.Spinal cord injury model produced in 42 Wistar rats on T7 by modified Allen impact method.Three days after injury,20 μl FSCS with a density of 1×105/μl prepared from pregnant rats (El4) in combination with AASCs were injected into the epicenter of the traumatized cavity.Animals were sacrificed at 2,4,6,8,10,12 weeks post transplantation.Light and electronmicroscopic studies as well as immunohistochemical assay were carried out to evaluate the graft survival,its differentation and integration with the host.ResultsIn the transplantation area,AASCs showed good growth and differentiation,and glial scarring surrounding the lesions was less.The neuroblast stretched out the terminal endings 4 weeks after implantation,followed by the presenting of the pre- and post-synaptic membrane.Eight weeks post transplantation,the dense or developed projections were observed in the pre- and post-synaptic membrane,the high electron dense substance full filled the synaptic cleft.All the spherical cleat vesicles,granular vesicles,elliptical vesicles and flattened-f type vesicles were discovered under the electron microscope.Ten weeks after injury,the axosomatic,dendrosomatic,dendro-dendritic,axoaxonic,and dendro-axonic synapses coexisted.Light microscopy showed that the graft cell grew gradually.Immunohistochemical assay showed that NF,5-HT,CGRP and GFAP positive fibers were in the graft.Synapses,glia fibers and blood brain barrier integrated each other.Conclusion1) The transplanted FSCS combined with AASCs can develop mature synapses with miscellaneous synaptic vesicles in the acute injured spinal cord.2) Co-existing indicate the possibility of synaptic connection between FSCS and host.

Objective To investigate the level of MEF2C phosphorylation (activation) and protein expression, and to further clarify the possible mechanism following ischemia-reperfusion in hippocampal CA1 region of rat. MethodsBrain ischemia was induced by four-vessel occlusion in SD rats. Protein level was determined by Western blotting. Results MEF2C was significantly activated with a peak at 6 h of reperfusion, but its protein expression decreased in late phase of reperfusion (3～5 d). The elevation of activated (17 ku) and the inactivated forms (32 ku) of caspase-3 proteases were remarkable during 1～5 d of reperfusion. In addition, Ac-DEVD-CHO, a specific inhibitor of caspase-3, up-regulated MEF2C protein level of 3 d reperfusion. SB202190 (an inhibitor of P38), but not ERK5-antisense oligonucleotides, not only inhibited MEF2C activation of 6 h reperfusion but also apparently prevented the increase of caspase-3 activation caused by 3 d reperfusion. Conclusion P38/caspase-3 mediated MEF2C pathway may function in the injuries of hippocampal CA1 region of rats following ischemia/reperfusion.

Objective:To explore the role of static posturography in the assessment of fatigue due to flight tasks.Methods:Thirtymale college students were asked to perform simulated flight tasks consecutively forfour hours.Meanwhile their statie posturography and tasks performance would be repeatedly measured during the task-load at end of every hour.Based on the changed significantly parameters,the static balance index would be built by principle component analysis.Then its correlation with task-load level would be further analyzed by curve estimation.Results:Static postural control declined significantly under effect of simulated flight tasks.With task load sustaining,static balance index increased significantly and correlated linearly with duration of task load (R2=0.949).Besides,there was quadratic relationship between the change of multi-tasks performance and duration of task load (R2=0.968).And correlation of multi-tasks performance with static standing balance level also had been proved to be quadratic (R2=0.976).Conclusions:Static posturography correlated linearly with flight task-load level,which could reflect fatigue level caused by task load.

Objective To apply Windows presentation foundation (WPF) to rapid access and report printing of medical data.Methods Data access was realized by abstracting database to form data structure through entity framework (EF) and mapping datasheet as attribute.Multipage printing was executed by using Flow Document Scroll Viewer as the container and putting contents into different paragraphs with the block element.Results EF of WPF decreased the frequency of database access by the program greatly,and the reaction speed of the program was increased significantly.Flow document could be used for data printing.Conclusion WPF enhances the efficiency of system development and the beauty of man-machine interface,and thus is worthy promoting for medical system and software development.

Objective To assess the validity of transanal local excision for stage Ⅰ low rectal carcinoma.Methods The clinical data of 93 patients with stage Ⅰ low rectal carcinoma who underwent transanal excision (group A,n=45)or radical resection(group B,n=48)were retrospectively analyzed.Twenty-four T1 patients and 21 T2 patients in group A received postoperative adjuvant radiation therapy and adjuvant chemoradiotherapy,respectively.All patients in group B received radical surgery only.The 5-year survival rates,recurrence rates,and postoperative complications between the 2 groups were compared.Results The 5-year survival rates were 100%(24/24)for T1 patients,86%(18/21)for T2 patients in group A,and 100%(18/18)for T1 patients,93%(28/30)for T2 patients in group B,with no significantly statistical difference between the 2 groups(P＞0.05).The recurrence rates were 4%(1/24)for T1 patients,19%(4/21)for T2 patients in group A,and 0(0/18)for T1 patients,7%(2/30)for T2 patients in group B,with no significance between the 2 groups(P＞0.05).The incidence of postoperative complications in group A was 2%(1/45),which was significantly lower than that of 15%(7/48))in group B(P＜0.05).Conclusions Transanal local excision of early low rectal carcinoma,combined with postoperative chemotherapy for T1 patients or chemoradiotherapy for T2 patients, results in a low complication rate and good sphincter function,and provides satisfactory local control and 5-year survival rates.

Objective To explore the neuroprotective role of Genistein (GEN) on hippocampal CA1 neurons and the possible mechanism following global cerebral ischemia ( GCI) in rats.Methods Seventy five rats were subjected to global cerebral ischemia ( GCI ) by four-vessel occlusion and randomly divided into five groups , sham, ischemia/reperfusion (I/R), GEN, ICI 182,780 and vehicle groups.Fluoro-Jade B and neuron-specific nuclear-binding protein ( NeuN) staining was used to observe CA 1 neuronal survival .TUNEL was used to detect apoptotic neurons .Spatial learning and memory function of the rats were evaluated by Morris water maze .Results The best dose of neuroprotective role of GEN was 1.0mg/kg body weight.Compared with sham, TUNEL-positive neurons in the hippocampal CA1 region increased significantly in I/R and vehicle groups (P<0.01), while post-treatment with GEN (1.0mg/kg) at 5min after ischemia by tail vein injection decreased markedly (P<0.01).Treatment of 1.0mg/kg GEN markedly attenuated spatial learning and memory deficits of the rats after ischemic insult compared to I /R group.Furthermore, ICI 182,780 significantly abolished the neuroprotective role of GEN (P <0.01).Conclusion The low-dose (1.0mg/kg) GEN significantly attenuates neuronal damage and cognitive deficits following GCI in rats , and the mechanism may be involved in estrogen receptor activity.

Objective To investigate the molecular mechanisms of protective effects of thioredoxin (Trx) on human vascular endothelial cells in atherosclerosis.Methods The cell models of Trx-overexpressing cells (Ad Trx) and the control cells (Ad-con) were established by adenovirus vector gene transfer technology in human umbilical vein endothelial cells (HUVECs).The oxidized low density lipoprotein,a risk factor of atherosclerosis,was used as a stimulator.Western blot and indirect immunofluorescence were used to detect the protein expression levels and the cellular localization of Trx,adhesion molecules (ICAM-1,VCAM-1) and the upstream signal pathways.Trx activity was detected by insulin disulfide reduction assay,and cellular reactive oxygen species (ROS)production was detected by fluorescent probe DCFH-DA.Results As compared with control group,Trx protein expression level was enhanced in Ad-trx group and the Trx activity in Ad-Trx group was upregulated by (26.2 ±3.3)％.The result of ROS detection showed that overexpression of Trx significantly inhibited the cellular ROS generation.As compared with control group,overexpression of Trx obviously inhibited the adhesion molecules expression but markedly promoted the phosphorylation of Smad3 in endothelial cells with or without oxLDL stimulation (P＜0.05).Pretreatment of cells with SIS3,a specific inhibitor of Smad3 phosphorylation,reversed Trx-induced inhibition of adhesion molecules expression.Further studies showed that pretreatment of cells with SIS3 enhanced oxLDL-induced AP-1 subunit c-fos nuclear expression.Conclusions The enhancement of Smad3 phosphorylation and c-Fos nuclear expression are mainly responsible for the Trx-induced downregulation of adhesion molecules.