Objective To investigate the changes ofplatelet parameters in type 2 diabetic patients with vascular complications.Methods One hundred and twenty-eight cases of type 2 diabetic patients were selected and divided into vascular disease group (49 cases) and non-vascular disease group (79 cases).Platelet count (PLT),mean platelet volume (MPV),mean platelet distribution width (PDW) and large-platelet (L-PLT) was detected,meanwhile 50 healthy volunteers were as control group.Results The MPV,PDW and L-PLT in vascular disease group was significantly higher than that in non-vascular disease group and control group [(13.98-2.35) fl vs.(11.53 ±2.06),(10.41 ± 1.95) fl; (15.87 ±2.18) fl vs.(13.51 ± 2.01),(11.15 ± 1.88) fl; (10.91 ± 1.34) × 109/L vs.(8.72 ± 1.03) × 109/L,(6.21 ± 0.98) × 109/L] (P ＜ 0.05),and PLT in vascular disease group was significantly lower than that in non-vascular disease group and control group [(115.2 ± 10.5) × 109/L vs.(129.6 ± 8.7) × 109/L,(176.5 ± 7.2) × 109/L] (P ＜ 0.05).Conclusion Platelet parameters detection are helpful for clinical diagnosis of diabetic angiopathy and play an important role in clinical diagnosis and disease monitoring.

Objective To.investigate the accuracy and reliability of monocyte count of three methods.Methods The specimens classified by blood cell analyzer WBC monocytes ＜ 15％,15％-20％,＞ 20％ were taken randomly,draw 20 μ l after mixing,put in monocyte direct count dilution and count,and underwent routine smear,stained by Swiss-Gibbs and manual classified by microscope,and then compared the results of the three methods.Collected one copy each of the high,medium and low proportion of monocytes specimens,and counted 30 times by the three counting method.Results Blood cell analyzer in WBC monocytes 15％-20％,＞ 20％ had poor correlation with the results of the other two methods.Repeatability comparison of three methods,medium and low sample were better with method 1 and 2,high value sample was good with method 3.Conclusion The specimens of monocytes ≥ 15％,especially scatter plot is abnormal,must be confirmed by microscopic examination.

Objective To establish animal models of diffuse axonal injury (DAI) and discuss the early functional MRI findings.Methods Twenty-five adult white rabbits were divided into control group (n =5),mild DAI group and severe DAI group (n =6) according to random number.Mild DAI group was subdivided into mild DAI 3 h (n =6),1 day (n =4) and 1 week (n =4) groups.Routine MRI and diffusion tensor imaging (DTI) were performed.Parameters measured were fractional anisotropy (FA),apparent diffusion coefficient (ADC),axial diffusivity (AD) and radical diffusivity (RD).Results Significantly increased FA (P＜0.01) and decreased ADC (P＜0.05) and RD (P＜0.01) were detected in severe DAI group compared to control group,but there was no significant variation in AD between the two groups (P ＞ 0.05).Each parameter remained almost unchanged in mild DAI 3 h group and mild DAI 1 week group,but FA was increased (P ＜ 0.01) and RD decreased (P ＜ 0.05) in mild DAI 1 day group,with no changes of ADC and AD.Conclusion DTI is a sensitive way to detect the occurrence and development of DAI and can provide referential images for early DAI diagnosis.

Objective To analyze the correlation between white matter injury and cognitive dysfunction using diffusion tensor imaging (DTI).Methods Seventeen subjects with TBI hospitalized from October 2012 to September 2013 had Glasgow coma scale (GCS) score of ≥ 13 (mild injury group, 10 cases) and ≤ 12 (moderate-severe injury group, 7 cases).Another 17 healthy subjects were used as controls.All were submitted to DTI examination.Fractional anisotropy (FA) and apparent diffusion coefficient(ADC) values in genu corpus callosum, splenium corpus callosum, posterior internal capsule, anterior internal capsule, and cerebral peduncle were calculated using the Neuro 3D software.Correlations between FA and ADC with the mini-mental state examination (MMSE) score were evaluated.Results Moderate-severe injury group demonstrated significantly reduced FA values in genu corpus callosum and splenium corpus callosum, and significantly increased ADC values of genu corpus callosum, splenium corpus callosum, posterior internal capsule and cerebral peduncle when compared to control group (P ＜0.05 or 0.01).FA and ADC values in the regions of interest did not differ significantly between mild injury group and control group (P ＞ 0.05).In the genu corpus callosum and splenium corpus callosum, FA values were positively correlated with MMSE score (r =0.636, 0.601), while ADC values were negatively correlated with MMSE score (r =0.552, 0.660).Conclusions DTI reveals the cerebral white matter lesion that is undetectable using CT and conventional MRI.DTI is a helpful tool to evaluate the degree of cognitive function in patients with TBI, which provides the basic reference for the clinical treatment and prognosis.

Objective To investigate the effect of electromyographic biofeedback motion sensing game on upper limb function in chil-dren with obstetric brachial plexus palsy (OBPP). Methods From March, 2013 to February, 2014, 41 children with OBPP were randomly as-signed into conventional rehabilitation group (n=21) and motion sensing game group (n=20). Both groups underwent a comprehensive course of rehabilitation, and the motion sensing game group received upper limb occupational therapy in the motion sensing game, for four weeks. They were assessed with Mallet shoulder function scale and the surface electromyogram (EMG) of the deltoid muscle when abduct-ing the shoulder before and after treatment. Results Before treatment, there was no significant difference in the Mallet scores between two groups (t=0.730, P>0.05), the scores improved after treatment in both groups (t>5.085, P<0.05), and the scores after treatment and the D-val-ue scores before and after treatment were higher in the motion sensing game group than in the conventional rehabilitation group (t>3.195, P<0.01). The integral EMG (iEMG) value, average EMG (aEMG) value and maximum EMG (EMGmax) value of the deltoid muscle improved in the motion sensing game group after treatment (t>4.420, P<0.05), however, only aEMG and iEMG improved in the conventional rehabili-tation group (t>2.282, P<0.05). All of them after treatment and the D-value were higher in the motion sensing game group than in the con-ventional rehabilitation group (t>2.155, P<0.05). Conclusion Electromyographic biofeedback motion sensing game therapy plays a role in the rehabilitation of upper limb function and muscle strength in children with OBPP.

The aim of surgical Chinese-English bilingual teaching is to improve medical students’ ability of foreign language and international intercommunication. Many methods are used to improve surgical Chinese-English bilingual teaching results,including understanding the importance,training persons qualified to teach,using and constructing English textbook,doing well examination,encouraging students to study and explore investigation of bilingual teaching.

Objective To investigate whether physical training can activate the PI3K/Akt (phosphatidyli-nositol 3-kinase/Protein Kinase B) signal transduction pathway after focal brain isehemia, leading to the reduction ofendothelial cell apoptosis. Methods Twenty-four male adult Sprague-Dawley rats (2 ～ 3 month old, n = 24) weresubjected to 60-min right middle cerebral artery occlusion (MCAO). All rats were randomly assigned to one of thethree groups: physical training group, control group and sham operation group. 24 hours after MCAO ,physical train-ing group underwent 30 min treadmill training per day for 2 weeks. Results After two weeks, the phosphorylationlevel of PI3K/Akt in the physical training group was significantly higher when compared with that in the control group(P <0.05), while the expression level of Bax in physical training group was lower when compared with that in thecontrol group( P < 0.05 ). Conclusion Physical training can activate PI3K/Akt signal transduction pathway,which may be associated with the neurological recovery.

Objective To determine the antigen-specific CTL in PBMC induced by a fusional family-gene vaccine of the immunoglobulin heavy chain variable gene framework region combined with the sequence of cytokine CM-CSF in vitro with MHC pentamer. Methods Peripheral blood samples were collected from two healthy donors and two patients. One was follicular lymphoma and another was hair cell leukemia. PBMC were isolated by density gradient centrifugalization with Ficoll and then subsequently differentiated into immature DCs (imDCs) induced by recombinant human GM-CSF and recombinant human IL-4. Gene gun was used to deliver the plasmids of the gene vaccine or the control plasmids into the imDCs. RT-PCR and ELISA assay were used to detect IgHVl-GM-CSF mRNA and GM-CSF in order to validate the transfection of the vaccine. After adding the cytokine cocktail, the imDCs became mature DCs. Then the mature DCs were co-cultured with lymphocytes from the blood samples for the induction of the antigen-specific CTL. The cultured cells were classified into vaccine group and control group and harvested at different time points of 0 d,7d, 17 d and 24 d after transfection. The subset of CD3+CD8+ T cells was analyzed by FCM assay. Finally, the CTL levels were detected with fluorescently labeled MHC pentamer antibody targeting vaccine epitopes. Results With the induction of cytokines, the imDCs with typical morphology were generated in PBMC. After delivering, the efficient expressions of the vaccine in the imDCs were determined by RT-PCR. And ELISA results also confirmed that GM-CSF was produced at a level of (28 ±6) ng/106 cells of the imDCs loaded with the vaccine, which was significantly different from that of control group (10 ± 3) ng/106 cells (t = 5. 191, P <0.01). FCM assay result showed that the CD3+ CD8+ T cells increased in a stepwise pattern during the culture. For control group, the levels at 0 d,7d, 17d and 24 d were ( 34. 24 ± 2. 72 )% , (46.06 ± 3.08)%, ( 65. 34 ± 4. 26 )% and (73.86 ±4.85 )% , respectively. For vaccine group, the results were (32. 28 ± 2. 08 ) % , (45. 32 ± 3. 81)% , ( 63. 37 ± 4. 21)% and (75. 01 ±3. 20)%. The differences between each time point had statistical significance (F = 176. 966 ,P <0.01) ,but there was no statistical differences between vaccine group and control group ( F = 0.657,P>0.05). The MHC pentamer analysis showed that the DCs loaded with IgHV1-GM-CSF fusional vaccine could efficiently induce the antigen-specific CTL response and the CTL levels increased gradually with the culture time, with the highest level of 4. 36% in the lymphoma blood and 3. 89% in the hair cell leukemia blood. Conclusions MHC pentamer assay could efficiently determine the antigen-specific CTLs response induced by the gene vaccine of family IgHV frame region in vitro. It could be a useful method for monitoring of anti-tumor cell immunity and evaluating of diagnosis and prognosis of the tumors in clinical application.

Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.

ABSTRACT:Objective To analyze the correlation between plasma nitric oxide (NO)level and atherosclerotic lesion in high cholesterol-fed (HCD ) rabbits.Methods Twenty male Japanese white rabbits were divided randomly into two groups,which were fed with normal diet (control group,n =10)or HCD (experimental group, n =10 )for 1 6 weeks.At the end of the experiment,plasma lipid and NO levels were measured.The gross atherosclerotic lesions in each group were detected by Sudan IV staining while intimal lesion area was measured by hematoxylin/eosin (HE)and elastica van gieson (EVG).Moreover,the macrophages (MΦ)and smooth muscle cells (SMC)were detected by immunohistochemical staining.The correlation analysis was made to reveal the relationship between atherosclerotic lesions and plasma NO level.Results Compared with those in control group, the total cholesterol (TC),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C)and NO levels all increased significantly in experimental group.Atherosclerotic lesions appeared on the vascular wall in the latter group.The area of atherosclerotic lesions and MΦ in the plaque had a positive association with plasma NO level. Conclusion There is a relationship between plasma NO level and the size of HCD-induced atherosclerotic lesions in rabbits.Meanwhile,the MΦ positive area in the atherosclerotic plaque is also associated with plasma NO level in cholesterol-fed rabbits,suggesting that plasma NO level may be associated with the occurrence and progress of early atherosclerosis.