Objective Through detecting the levels of von willebrand factor (vWF) ,vWF-cleaving protease (vWF-cp)before and after percataneous coronary intervention (PCI) in patients with coronary atherosclerotic heart disease ,to evaluate the relationship between them .Methods According to the results of coronary angiography ,study objects were divided into control group (normal) and PCI group ( the diagnosis of coronary heart disease with PCI operation ) .Enzyme-linked immunosorbent assay (ELISA) was carrout to determin the levels of plasma vWF concentration and vWF-cp activity in plasma and statistical analysis .Results The pre-operative ,postoperative vWF levels in plasma in PCI group were significantly higher than that in the normal control group ;The plasma vWF levels after PCI were significantly higher than that in the group before PCI (P< 0 .05) ;vWF-cp level of the PCI group were lower than in the control group ,and postoperation is lower than preoperation ( P < 0 .05) .With the coronary artery lesion worsen ,the plasma level of vWF increased ,while the level of vWF-cp activity decreased .Conclusion vWF and vWF-cp levels with the severity of coronary atherosclerosis and thrombosis risk was increased in different decreased ,which may play an important role in the pathogenesis of acute coronary syndromes .Changes of vWF and vWF-cp level after PCI indicate that interventional therapy can increase the risk of thrombosis to some extent .

BACKGROUND:Streptozotocin-induced diabetic ophthalmopathy is commonly used in animal models, but the pathological changes are local that mainly emphasize on the retina. Little evidence is found about the animal models of the pathology of diabetic ophthalmopathy. OBJECTIVE:To investigate the long-term stability of type 2 diabetic melitus rat models induced by streptozotocin combined with high-fat feeding and to observe the characteristics of eye disease. METHODS: Rats were randomly divided into control group and diabetes group. Control group was given normal feeding, while diabetes group was given intraperitoneal injection of streptozotocin combined with high-fat feeding to establish diabetic models. RESULTS AND CONCLUSION:Compared with the control group, at 1 month after modeling, the fasting blood glucose levels increased, and the insulin sensitivity index decreased in the diabetic group (P < 0.05). Evans blue staining results showed that, at 3 months after modeling, retinal cel lesions exacerbated in the diabetic group; at 5 months after modeling, retinal blood vessels traveled in circuity and disorderly, accompanied by the leakage in the diabetic group, Evans blue content in the retina increased as the time after modeling went by (P < 0.05). Under transmission electron microscopy, at 5 months after modeling, the eye lenses in the diabetes group were flocculent pieces, which were the typical characteristics of cataract. Experimental findings indicate that the rat model of type 2 diabetic melitus induced by streptozotocin combined with high-fat feeding has a long-term stability, and its eye changes are consistent with the characteristics of diabetic ophthalmopathy. Therefore, it is an ideal animal model for diabetic ophthalmopathy.

Objective:To observe the up-regulation of nuclear Clusterin (nCLU)gene on the biological behaviors of human non-small cell lung cancer cell line A549 .Methods:Sense eukaryotic expression vector of nCLU was constructed by cloning the cDNA of nCLU into pIREShyg3 vector. A549 cells were transfected with pIRES-nCLU and pIREShyg3 vectors by lipofectAMINE~(TM) 2000 mediation, respectively. Stable transfected cells were selected by hygromycin B screening. CCK-8 assay was performed to evaluate the effect of nCLU over-expression on cell proliferation in vitro. The expression level of nGLU protein was examined by Western blotting. Cell cycle distribution was detected by FCM with PI staining. The alteration of migration and metastasis potential of A549 cells before and after nCLU gene transfection were assayed by cell chemotactic migration and invasion test. Results:The proliferation speed of the transfected A549 cell clones stably over-expressing nCLU was slowed down. FCM analyses revealed that the percentage of cells in G_0/G_1 phase dramatically increased from (33.54±2.10)% to (63.31±4.30)%. The cell chemotactic migration and invasion potentials were markedly reduced after nCLU gene transfection (P<0.05). Conclusion:Up-regulation of nCLU can greatly inhibited the proliferation and decreased the migration and invasion capabilities of A549 cells.

Aim To investigate the effect of phenyle-thanol glycosides from Cistanche tubulosa(CPhGs) on the proliferation and activation of rrPDGF-BB induced HSC and their target points for resisting hepatic fibro-sis,to elucidate the molecular mechanism in molecular level, and provide basic data for the further develop-ment of new drugs. Methods HSCs were cultivated by CPhGs with different concentrations ( 0 , 3. 91 , 7. 81 , 15. 63 , 31. 25 , 62. 50 , 125. 00 , 250. 00 , and 500 mg ·L-1 ) and IC50 of CPhGs was determined. CPhGs with different concentrations ( 25 , 50 , 75 , 100 mg · L-1 ) were selected, and after the cells were stimulated with rrPDGF-BB, cell proliferation was determined by MTT. ERK1/2 ,α-SMA, c-fos, c-jun and Collagen I mRNA and Erk1/2 ,P-Erk1/2 and CollagenⅠprotein ex-pressions were assayed by RT-PCR and Western blot. Results CPhGs of ( 50 ~100 ) mg · L-1 concentra-tions groups could effectively inhibit rrPDGF-BB-medi-ated proliferation(P<0. 05) and CPhGs of(25~100) mg·L-1 concentrations groups had no significant cyto-toxicity( P >0. 05 ) . CPhGs of ( 25 ~100 ) mg · L-1 concentrations groups could inhibit ERK1/2 ,α-SMA,c-fos, c-jun and CollagenⅠmRNA levels, and also ob-viously inhibited Erk1/2 ,P-Erk1/2 and Collagen Ⅰ pro-tein expression on HSC. Conclusions CPhGs has the protective effect against hepatic fibrosis. The mecha-nism of this process may involve the interference with PDGF/ERK1/2 signaling pathway and inhibiting the activation and proliferation of HSC.

Objective To explore whether Cdc42 is an independent influence factor in regulating the migration of A549 cells by using quantitative measurement method,and to verify the effectiveness of an automatic measurement and calculation method for in vitro cell migration assay.Methods Cdc42 was overexpressed in human lung adenocarcinoma A549 cell line,and then in vitro scratch assay was applied to evaluate the migration of the cells.Different methods were used to measure the images acquired at different time points for quantitative analysis.Accuracy and repeatability of different measurement methods were analyzed.Results The results showed that overexpression of Cdc42 alone significantly advanced the migration of A549 cells (P＜0.05).The new method is efficient,accurate and reproducible as compared to manual measurement,and has significant advantages in target recognition and noise removal as compared to the existing measurement method in Image Pro Plus 6.0.Conclusions Over expression of Cdc42 significantly increased A549 cell migration ability in vitro.The new method can realize the automatic quantitative analysis of cell migration in vitro.

Objective To compare the outcomes between modified McKeown minimally invasive approach and open left chest-neck incision approach esophagectomy for treatment cancer of mid-to-distal thoracic esophagus.Methods We retrospectively analyzed clinical data from 128 patients with mid-to-distal thoracic esophageal cancer who underwent thoracoscopic and laparoscopic esophagectomy from March 2009 to March 2012.One hundred and fifty patients were served as control that underwent open left chest-neck incision approach esophagectomy in the same period.Results All the operations were performed successfully.There was significant difference between modified McKeown minimally invasive approach and open left chest-neck incision approach esophagectomy with regard to respiratory complications (10.9 ％ vs.20.7％),pneumonia (4.7％ vs.11.3 ％),atelectasis (3.1％ vs.10.5 ％,),pleural effusion (3.1％ vs.10.0％) and delayed gastric emptying (8.6 ％ vs.1.3 ％) (P ＜ 0.05).Hospital stay was significantly shorter in the minimally-invasive group than the open group [(11.7 ± 3.6) days vs.(13.9 ± 6.5) days,P＜0.05],and had significantly less blood loss [(88.1 ±41.8) ml vs.(360.5 ±80.6) ml,P＜0.05] and the number of lymph nodes harvested (22.9 ±5.7 vs.16.8 ±4.5,P ＜0.05).No significant differences were observed on the operative time,mortality and other complication between the two groups.Conclusion Modified McKeown minimally invasive approach esophagectomy is techeniqually feasible and safely which have lower blood loss,lower respiratory complication,shorter hospital stay and more number of lymph nodes harvested comparing to open left chest-neck incision approach.

Objective To analyze the pattern of normal bone marrow on whole body diffusionweighted imaging (WB-DWI) and its influence factors.MethodsA total of 98 healthy volunteers (male 47 and female 51). All volunteers underwent WB-DWI at 1.5 T MR scan. The ADC value,the signal intensity on DWI obtained with a b value of 800 s/mm2 ( SIDWI ),and the signal intensity on short time inversion recovery images ( SISTIR ) of thoracic vertebrae,lumber vertebrae,bilateral head of femur,bilateral neck of femur,bilateral superior segment of femur,bilateral inferior segment of femur,bilateral ilium,bilateral head of humerus,bilateral scapula were measured and compared with ANOVA test and StudentNewmar-Keuls test.The normal appearance of bone marrow on WB-DW[ was assessed. The relationship between the SIDwI and the ADC,and the SIDWI and the SISTIR of bone marrow were analyzed.The ADC of bone marrow between male and femaIe were compared. Spearman correlation analysis was performed for different age groups.Results( 1 ) Bone marrow signal intensity was different among 98 healthy volunteers.Bone marrow in 69 healthy volunteers (female 24,male 45 ) showed low to intermediate signal intensity,whereas in the remaining 29 healthy volunteers (female 27,male 2) showed high signal intensity.(2) The SIDWI of thoracic vertebrae ( median 44.54),lumber vertebrae ( median 35.01 ),head of femur ( median 13.61 ),neck of femur ( median 16.00),superior segment of femur ( median 21.45 ),ilium ( median 25.77),head of humerus (median 18.35),scapula (median 36.12) was positively correlated with the ADC [ (0.55 ±0.08) × l0-3,(0.53 ±0.08) × 10-3,(0.30 ± 0.10) × 10-3,(0.42 ± 0.16) × 10-3,(0.74±0.14) ×l0- 3,(0.49±0.10) ×10-3,(0.36±0.13) ×10-3,(0.49±0.11) × 10-3mm2/s]and the SISTIR ( median 61.81,64.99,53.27,69.08,73.10,66.35,73.16,79.81 ),r =0.513 and 0.695,0.741 and 0.764,0.443 and 0.489,0.641 and 0.656,0.510 and 0.648,0.475 and 0.715,0.366 and 0.446,0.437 and 0.739 ;P ＜ 0.01. (3) There was significant difference of the ADC of bone marrow in different bone,F =138.69,P ＜ 0.01. Student-Newman-Keuls test revealed that no significant difference was found in the ADC between thoracic vertebrae and lumbar vertebrae,ilium and scapula,head of humerus and inferior segment of femur ( P ＞ 0.05 ),and significant difference was found in the ADC values between the remaining two groups ( P ＜ 0.05 ). The bones associated with decreasing ADC values were superior segment of femur,thoracic vertebrae and lumber vertebrae,ilium and scapula,neck of femur,head of humerus,head of femur and inferior segment of femur. ( 4 ) The ADC values of bone marrow of female subjects in thoracic vertebrae [ (0.59 ±0.07) × 10-3 mm2/s],lumber vertebrae [ (0.58 ±0.06) × 10 -3 mm2/s],head of femur ( median 0.33 × 10 -3 mm2/s),neck of femur ( median 0.53 × 10 -3 mm2/s),superior segment of femur ( median 0.81 × 10-3 mm2/s),inferior segment of femur ( median 0.32 ×10-3 mm2/s),ilium [ (0.52 ± 0.09 ) × 10-3 mm2/s ],head of humerus (median 0.42 × 10-3 mm2/s),scapula [ (0.53 ± 0.09) × 10-3 mm2/s] were significantly higher than those of male subjects [ (0.51 ±0.07) × 10-3,(0.48 ±0.07) × 10-3,median 0.23 × 10-3,median 0.31 × 10-3,median 0.66 × 10-3,median 0.23 × 10-3,(0.46 ±0.10) × 10-3,median 0.27 × 10-3,(0.45 ±0.11 ) × 10 3mm2/s].(5)There was significant negative correlation between the ADC values of bone marrow and age in thoracic vertebrae,lumber vertebrae,head of femur,neck of femur,superior segment of femur,ilium,head of humerus for female subjects,r =-0.549, -0.629, -0.329, -0.524, -0.338, -0.548 and -0.416,respectively,P ＜ 0.05.There was no significant correlation between ADC values and age in inferior segment of femur and scapula for female subjects and all the regions for male subjects ( P ＞ 0.05 ).Conclusions The ADC and the SIsTIR of bone marrow correlates with the SIDW1.The ADC values of bone marrow is affected by age and sex,and is different for different bones.

BACKGROUND:Previous studies have shown that kainic acid injected into hippocampus can significantly upregulate the expression of excitatory KA1 subunit of the kainate receptor in the hippocampus, and endoplasmic reticulum stress markers, phosphorylation of the alpha subunit of eukaryotic initiation factor 2, accompanied by celldeath. OBJECTIVE:To explore the mechanism of endoplasmic reticulum stress after kainic acid is injected into the hippocampus.METHODS:0.15 nmol kainic acid was injected into the hippocampal CA1 region of 32 adult male Kunming mice, the injection time was 60 seconds. At different time points (1, 2, 3, 4, 5, 6, 8 and 12 hours) after kainic acid was injected, the Bederson score analysis was performed, and then the brain was harvested after cerebral perfusion. FJB staining of brain sections and immunofluorescence double labeled observation were also performed. RESULTS AND CONCLUSION:(1) At 3, 4, 5, 6, 8 hours after kainic acid injection, Bederson score showed severe injury of central nervous system function, and FJB staining showed the increased of celldeath in the hippocampus (P<0.05);At 1, 2, 12 hours after injection, Bederson score showed no obvious injury of central nervous system function, and FJB staining showed unobvious celldeath in the hippocampus (P>0.05). (2) According to the results of FJB staining, the brain sections were selected at 3, 8 hours for immunohistochemistry. The expressionlevels of KA1 receptors and endoplasmic reticulum stress marker P-eIF2αwere up-regulated at the same time after kainic acid was injected into hippocampus. Two single-staining KA1 and P-eIF2αimmunofluorescence images were synthesized into one over-lapped double-stained image, and two images overlapped, indicating that the up-regulated expression of KA1 and endoplasmic reticulum stress occurred in the same nerve cells. Kainic acid first up-regulated the excitatory receptor KA1 expression, which may cause cellendoplasmic reticulum dysfunction and result in the endoplasmic reticulum stress response, further promoting neuronal celldeath.

Objective To investigate the changes of CD4~+ T cell subset in the role of immuno-pathogenesis of type 1 diabetes mellitus(T1DM). Methods Real-time PCR was used to evaluate the ex-pression levels of transcriptional factors (T-bet, GATA-3, Foxp3, ROR-γt), cytokines (IFN-γ, IL-4, IL-10, IL-17A) and negative regulatory factors (CTLA-4, GITR) mRNA from CD4~+ T cells. The proportions of Th1, Th2, Tr and Th17 cells in peripheral blood were detected by flow cytometric analysis. Plasma cyto-kine (IFN-γ, IL-4, TGF-β and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) The proportions of Th1 cells in peripheral blood from children with T1DM were siguificanfly increased than that of healthy controls, and proportions of Th2 were decreased (P < 0.01). There were no significant differences between diabetic patients and healthy controls regarding the proportions of Tr cells and Th17 cells(P >0.05). (2) Transcription levels of T-bet and IFN-γ mRNA were significantly up-regulated, while GATA3 and IL-4 were significantly down-regulated in children with T1DM. The mRNA expression levels of Tr negativity regulatory factors such as IL-10, CTLA-4 and GITR were lower in CD4~+ T cells from children with TIDM compared with the controls(P <0.01). There were no statistically differences to be observed in mRNA expression levels of ROR-γt and IL-17A genes between two groups(P > 0.05).(3) In comparison with controls, serum concentrations of IFN-γ or IL-4 were remarkable increased or de-creased respectively (P < 0. 01), while TGF-β and IL-6 did not change in children with T1DM (P > 0.05). Conclusion The Th1/Th2 imbalance might be play an important role in immunopathogenesis of T1DM. Functional deficiency of Tr cell might further exacerbate Th1/Th2 imbalance and lead to disturbance of cellu-lar immune response.