AIM: To verify the neuroprotective effect and optimize the therapeutic dose and time window of picroside Ⅱon cerebral ischemic injury in rats .METHODS:The forebrain ischemia model was established by the method of bilateral common carotid artery occlusion ( BCCAO ) .The successful model rats were randomly divided into 16 groups according to orthogonal design and treated by intraperitoneal injection of picroside Ⅱat different ischemic time poinis and different doses .The changes of the nerve fiber myelin were observed by fast green staining .The immunohistochemical assay and Western blotting were used to quantitatively and qualitatively determine the expression of myelin basic protein (MBP). The mRNA level of MBP in the brain tissues was tested by reverse transcription polymerase chain reaction (RT-PCR).RE-SULTS:Picroside Ⅱ increased the expression of MBP and decreased demyelination after cerebral ischemic injury .The best therapeutic time window and dose were:(1) ischemia for 2.0 h with picrosideⅡat dose of 10 mg/kg according to the results of fast green staining;(2) ischemia for 2.0 h with the dose of 10 mg/kg according to the results of immunohisto-chemical assay;(3) ischemia for 2.0 h with the dose of 10 mg/kg according to the analysis of Western blotting;(4) is-chemia for 1.5 h with the dose of 20 mg/kg according to the detection of RT-PCR.CONCLUSION:Given the principle of the lowest therapeutic dose with the longest time window , the optimized therapeutic dose and time window for rat cerebral ischemic injury is intraperitoneal injection of picroside Ⅱat the doses of 10~20 mg/kg and the time window of ischemia for 1.5~2.0 h.

In order to fit new plot proposed by medical education reform,by integrating interrelated resources,the author introduced peer education into the whole process of nutrition propa-ganda and education to promote the development of the whole social health education.

Objective To study the association between CYP2C9 gene polymorphism and warfarin maintenance dosage in anticoagulation therapy.Methods 200 Han patients admitted to our hospital for heart valve replacement were included in this study.CYP2C9 * 2,CYP2C9 * 3,CYP2C9 *c65 in CYP2C9 gene were sequenced using the CAPS technique and conventional DNA sequencing method.Dosages of warfarin used in patients carrying different genes were analyzed.Results No mutation of CYP2C9 * 2 but only one kind of allele C was detected in 200 patients.The genotype of CYP2C9 * 2 was C/C wild type.Allelic gene was detected at CYP2C9 * 3 A and C,with A/A wild type detected in 171 patients,A/C heterozygote mutation type detected in 18 patients,and C/C heterozygote mutation type detected in 11 patients respectively.The frequency of allelic genes A and B was 94.3 ％ and 5.7 ％ respectively.A significant difference was found between CYP2C9 * 3 mutation and warfarin dosage (P＜0.05).The dosage of warfarin reduced 18.46％ and 76.0％ respectively in patients carrying A/C heterozygote mutation type and in those carrying C/C heterozygote mutation type.Two kinds of allelic gene were detected at CYP2C9 * c65 G and C,with G/G wild type detected in 182 patients and G/C heterozygote mutation type detected in 18 patients respectively.No significant association was found in warfarin maintenance dosage for patients carrying G/G wild type and G/C heterozygote mutation type.Conclusion CYP2C9 gene polymorphism is associated with warfarin maintenance dosage in anticoagulation therapy.

Objective; To investigate the stress distribution in implant-bone interface with different pitches by three-dimension finite element analysis method in order to decide the best design of thread parameters for stress distribution. Methods: Built three-dimension finite element models of partial mandible and dental implant with superstructure. The stress distributions in bone and three kinds of implants whose pitches were 0.6, 0.8 and 1.0 mm respectively, under axial and oblique 45 degree to axle concentrated load. Data were analyzed by Cosmos/works software. Results;The maximal Von-Mises stress, tensile stress and compressive stress of implant with 0.8 mm pitch were lower than others, the stress distribution was even. The stresses in the same model under oblique load were much higher than those under axial load. The stress concentrated more on implant neck, upper part of cortical bone and the lowest thread on implant. Conclusion; The pitch of implant has effect on stress distribution in implant-bone interface and the transmission of occlusal force. The lower threads on implant should be cut to avoid the concentration of stress, and the non-axial force should be reduced as possible as we can or avoided when we design and make implant denture.

Objective To explore the characteristic findings of 1H-MR spectroscopy(1H-MRS) in the prefrontal cortex and amygdala of patients with heroine dependence(HD), and the relationship to total cumulative dose of inhaled heroine.Methods Fourteen male HD patients and 12 healthy controls (HC) underwent 1H-MRS at the prefrontal cortex and amygdala regions.The total cumulative in haled heroin dose was (852±341) g in HD.Ratios of N-acetylaspartate/creatine(NAA/Cr) and choline/creatine (Cho/Cr) were respectively measured in the prefrontal cortex and bilateral amygdale regions.The student's t test and the linear correlation were employed for statistical analysis.Results Compared to HC group, HD patients had a significant lower ratio of NAA/Cr in the prefrontal cortex (1.44±0.46 vs 1.50±0.75, t=1.77 ,P <0.05), left amygdala region (1.32±0.08 vs 1.42±0.08, t=3.41, P < 0.05), and right amygdala region (1.34±0.09 vs 1.44±0.10, t=2.63, P <0.05), the HD patients had a significant increased ratio of Cho/Cr in the prefrontal cortex (0.92±0.06 vs 0.86±0.08, t=2.31, P < 0.05), left amygdala region (1.20+0.12 vs 1.07±0.04, t=3.60,P<0.05) and right amygdala region(1.26±0.15 vs 1.12±0.11,t=2.60,P <0.05).There was a negative linear correlation between the total cumulative inhaled heroine dose and the ratio of NKA/Cr in the prefrontal cortex (r=-0.9159, P < 0.01), left amygdala region (r=-0.8756, P < 0.01), and right amygdala region (r=-0.9399, P < 0.01) respectively.Conclusions The study indicates that neuronal damage and glial proliferation may occur in the prefrontal cortex and amygdala region, which suggests the abnormalities of executive function and emotion in patients with HD.A relationship exists between the heroin-induced metabolic abnormality and the total cumulative dose of inhaled heroine.

Objective To determine the changes of peripheral levels of T helper cell cytokines of patients with chronic hepatitis B (CHB) during antiviral treatment,and to further explore its clinical significance.Methods The plasma levels of interleukin (IL)-2,IL-6,IL-10,interferon (IFN)-γ and tumor necrosis factor(TNF)-α of thirty-three CHB patients during antiviral treatment (entecavir) were measured using enzyme linked immunosorbont assay (ELISA).And their biochemical indicators of liver function were determined.The differences of cytokines levels before and after antiviral treatment were compared using ANOVA.The correlations between the changes of cytokines and alanine transaminase (ALT),hepatitis B virus (HBV) DNA levels were analyzed.Results Levels of IFNγ before and 12,24,48 weeks after treatment were (5.98±2.77),(5.95±3.37),(2.93±2.15) and (9.29±4.65) pg/mL,respectively (F=3.845,P＜0.05),which were positively correlated with ALT levels (r =0.396,P＜0.05).Both TNF-α and IL-10 levels declined after antiviral treatment,which were significantly different at different time points (F=20.156 and 16.695,respectively; both P＜0.05),and both levels of TNF-α and IL-10 were positively correlated with ALT levels (r=0.354and 0.316,respectively; both P＜0.05) and positively correlated with HBV DNA levels (r=0.382and 0.386,respectively; both P＜0.05).While both IL-2 and IL-6 levels were not significantly different between before and after antiviral treatment (F=0.010 and 0.932,respectively; both P＞0.05).The serum levels of ALT and HBV DNA before and after antiviral treatment were all significantly different (F=17.69 and 198.98,respectively; both P＜0.05),which declined gradually during treatment and were positively correlated (r =0.581,P＜0.05).Conclusions IL-10,IFNγand TNF-α may be involved in the pathologic process of CHB,and closely related to the deterioration of the disease.Monitoring plasma levels of these cytokines during antiviral treatment could be useful to understand the immune status and evaluate the efficacy of antiviral drugs.

Organ system based integrated teaching model has been adopted in several medical schools, and these schools face some challenges in this teaching process. In order to provide new sights for organ system based integrated teaching reform in eight-year medical education program, ex-ploration of setting up three-staged-dimensional integrated teaching models was conducted in urinary system by hematuria, and designing related teaching objectives, teaching programs and assessment form to optimize teaching quality in this teaching methods, according to characteristics of the urinary systemic diseases, cultivating objectives of eight-year medical education program and teaching experi-ence of Xiangya School of Medicine.

The present paper is aimed to study the preparation and application of individual artificial bone of carbon/carbon composites. Using computer tomography images (CT), we acquired a three-dimensional image. Firstly, we described bone contour line outlined with manual and automatic method by the binary volume data. Secondly, we created 3D object surface information by marching cubes. Finally, we converted this information to non-uniform rational B-spine (NURBS) by using geomagic software. Individual artificial bone with carbon/carbon composite was prepared through the CNC Machining Center. We replaced the humeral head of the tested rabbit, and then observed the effects of implantation in neuroimaging and pathological section. Using this method, we found that the bone shape processed and bone shape replaced was consistent. After implantation, the implant and the surrounding bone tissue bound closely, and bone tissue grew well on the surface of the implant. It has laid a sound foundation of the preparation using this method for individual artificial bone of carbon/carbon composite material.
Animals ; Bone Substitutes ; chemistry ; Carbon ; chemistry ; Imaging, Three-Dimensional ; Rabbits ; Software ; Tomography, X-Ray Computed

Objective To observe the effect of the three active ingredients of a Chinese traditional medicine compound named Kang Fu Ling( KFL) against PC12 cells oxidative damage induced by microwave radiation.Methods PC12 cells were differentiated into neuros induced by nerve growth factor ( NGF ) .PC12 cells were incubated for 48 hours after astragalosides,total paeony glycoside and tanshinones were added at different concentrations (1, 3, or 9 μg/ml) .The cells in the control group were cultivated with the only medium of the same volume.Then, cells were irradiated with 30 mW/cm2 microwave for 6 minutes.The morphology of PC12 cells was observed under an inverted microscope soon before and after irradiation and the cell viability was measured by methylthiazolyl tetrazolium ( MTT) colorimetry.Reactive oxygen species ( ROS ) was determined using active oxygen probe 2′, 7′-dichlorodihyarofluolescen diacetde ( DCFH-DA ) while malonyldialdehyde(MDA) was measured in the homogenate of PC12 cells through thiobarbituric acid ( TBA) reactive substance assay.Results The cell morphology of each group showed no obvious difference.6 h after irradiation, the viability of irradiation control group measured by MTT declined apparently(P<0.01)compared with the normal control group.The 3 μg/ml astragalosides treatment group increased the viability of PC12 cells after microwave exposure ( P <0.01).The contents of ROS and MDA were increased after irradiation(P<0.01).However, in the three active ingredients of Kang Fu Ling treatment groups, both ROS and MDA were much lower than in irradiation control group.Conclusion Astragalosides, total paeony glycoside and tanshinones, which are the three active ingredients of Kang Fu Ling, all have protective effect against PC12 cell injury caused by microwave radiation,possibly by scavenging free radicals and reducing oxidative stress injury.

OBJECTIVETo prepare stable chicken red blood cells for the calibration of flow cytometry.

METHODSThe traditional isolation method of chicken red blood cells was modified by incorporating gelatin technique, Ca2+-free HBSS treatment and low-speed centrifugation. The effect of fluorescence staining of the cells was improved by the addition of TritonX-100 to enhance the membrane permeability and Rnase enzymes to disintegrate RNA tiles. The modified method was compared with the traditional method for viability of the freshly isolated cells and the DNA content coefficient of variation (CV) of the fixed cells.

RESULTSChicken red blood cells obtained by the modified method showed a significantly higher viability than those obtained by the traditional method [(98.5∓3.5)% vs (93.5∓2.7)%, P<0.05]. After glutaraldehyde fixation, the isolated cells with the modified method were stable during the 90-day preservation with a significantly lower CV than the cells obtained by the traditional method [(6.0∓0.3)% to 6.2∓0.4% vs (8.6∓0.5)% to (13.1∓1.4)%, P<0.01].

CONCLUSIONThe chicken red blood cells isolated using the modified method can be applicable for calibration of flow cytometry.