Background Experimental study showed that heparanase (HPA)is overexpressed in choroidal neovascularization,suggesting that it may play a role in the pathogenesis of angiogenesis.To certify HPA inhibitor suppress the formation and development of new blood vessel has an important significance for the treatment of choroidal neovascularization.Objective The aim of this study was to investigate the effect of HPA inhibitor on the proliferation of human umbilical vein vascular endothelial cell (UVEC) and the expression of HPA.Methods Hunan UVEC was primarily cultured and passaged and the third generation cells were used in the experiment.Phosphomannopentaose sulfate (PI-88) solution,a HPA inhibitor,was prepared with endothelial cell medium and the end concentrations were 400.00,200.00,100.00,50.00,25.00,12.50,6.25 mg/L respectively.The cells were treated with PI-88 solutions for 24,48 and 72 hours.The growth and proliferation of human UVEC were analyzed using MTT colorimetric assay at absorbance 570 nm.The expression of HPA in the cells was detected by immunochemistry in 48 hours after addition of PI-88.Results Cultured human UVEC showed the fusiform and polygon in shape.The A570 values of human CVEC were significantly different among various concentrations of PI-88 groups (F=2.721,P=0.053 ) and different action time (F=9.656,P =0.002).When PI-88 was administered for 24 hours,the A570 values of human UVEC were insignificantly altered in comparison with the one without PI-88 culture group (P＞0.05 ).However,in 72 hours after experiment,the A570 values were significantly declined as the PI-88 concentration was ＞50.00 mg/L ( P＜0.05 ).When PI-88 was administered for 48 and 72 hours,the A570 values of human UVEC were significantly higher than those of 24 hours in ＜50.00 mg/L groups (P＜0.01 ),but no statistical differences were seen in ＞100.00 mg/L groups among various time points (P＞0.05 ).HPA was intently expressed in the cytoplasm of human UVEC.However,at 48 hours after addition of ＞25.00 mg/L PI-88,the HPA expression was obviously weaker.Conclusions PI-88 can suppress the growth and proliferation of human UVEC at the dose-and time-dependent manner by downregulating the expression of HPA in the cells.

ATP-binding cassette transporter Al (ABCAl) is a kind of membrane intergrate protein and may have multiple and diverse functions. It mediates the cellular efflux of phospholipids and cholesterol to lipid-poor apolipoproteinA- Ⅰ (apoA- Ⅰ ) and plays a significant role in high density lipoprotein (HDL) metabolism. Mutations in human ABCAl cause severe HDL deficiencies characterized by the virtual absence of apoA- Ⅰ and HDL and prevalent atherosclerosis. ABCAl expression is highly regulated and implies a variety of molecular actors. All of the nuclear receptors which involve in regulation of ABCAl expression act via the DR4 element in the ABCAl promoter. cAMP up-regulates ABCAl expression by acting both at the transcriptional and translational level. Cytokines have been shown to exert pleiotropic and antinomic effects on ABCAl transcription, In addition to these, some of enzymes and proteins such as protein kinase A, protein kinase CK2, cathepsin D are involved in the regulation of ABCAl expression. The recent progress in the structure, function and regulation of ABCAl transporter is reviewed.

OBJECTIVE: To investigate the lethal blood level, the target organs and tissues, the toxicant storage depots and the postmortem redistribution in mice died of emamectin benzoate poisoning. METHODS: The mice model of emamectin benzoate poisoning was established via intragastric injection. The main poisoning symptoms and the clinical death times of mice were observed and recorded dynamically in the acute poisoning group as well as the sub-acute poisoning death group. The pathological and histomorphological changes of organs and tissues were observed after poisoning death. The biodistribution and postmortem redistribution of emamectin benzoate in the organs and tissues of mice were assayed by the enzyme-linked immunosorbent assay (ELISA) at 0h, 24h, 48h and 72h after death. The lethal blood concentrations and the concentrations of emamectin benzoate were detected by high performance liquid chromatography (HPLC) at different time points after death. RESULTS: The symptoms of nervous and respiratory system were observed within 15-30 min after intragastric injection. The average time of death was (45.8 ± 7.9) min in the acute poisoning group and (8.0 ± 1.4) d in the sub-acute poisoning group, respectively. The range of acute lethal blood level was 447.164 0-524.463 5 mg/L. The pathological changes of the organs and tissues were observed via light microscope and immunofluorescence microscope. The changes of emamectin benzoate content in the blood, heart, liver, spleen, lung, kidney and brain of poisoning mice showed regularity within 72 h after death (P < 0.05). CONCLUSION The target organs of emamectin benzoate poisoning include heart, liver, kidney, lung, brain and contact position (stomach). The toxicant storage depots are kidney and liver. There is emamectin benzoate postmortem redistribution in mice.
Animals ; Autopsy ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Ivermectin/toxicity* ; Lethal Dose 50 ; Mice ; Postmortem Changes ; Tissue Distribution

Objective To identify etiological factors for non-ulcer dyspepsia (NUD) in elderly patients presenting to the emergency department.Methods A total of 97 elderly patients with NUD presenting to the emergency department from January 2011 to December 2013 were enrolled in the NUD group and 116 elderly people undergoing annual physical examinations served as the control group.Clinical data,including Helicobacter pylori (HP) infection,unhealthy living habits,anxiety,depression and regular intake,were collected and compared between the two groups.Results Compared with the control group,the NUP group showed a higher HP infection rate [70.1％ (68/97)vs.54.3％ (63/116),x2 =5.565,P＜0.05],a higher incidence of unhealthy living habits (unhealthy dietary habits,smoking,drinking,and lack of regular exercise) (P＜0.05),a higher incidence of suspect anxiety/anxiety and a higher incidence of regular intake [30.9％ (30/97) vs 10.3％ (12/116),x2=14.138,P＜0.05].SerumpepsinogenⅠ(PGⅠ) [(178.7±13.6) μg/L vs.(125.2±10.5)μg/L,t=1.971,P＜0.01] and serum pepsinogen Ⅱ(PG Ⅱ) [(28.8±5.3) μg/L vs.(14.7±3.8)μg/L,t=1.960,P＜0.01] levels were also higher in the elderly NUD patients than in the control group.Conclusions HP infection,unhealthy living habits,anxiety and/or depression,and regular intake are closely associated with NUD in the elderly.Serum PGⅠ and PGⅡ could be used as markers in screening for malignant lesions in elderly patients with dyspepsia.

Objective To investigate the effects of p38 mitogen-activated protein kinase (MAPK) short hair RNA (shRNA) delivered by lentiviral vectors (pGLV) on cardiac function after myocardial infarction (MI) in aldosterone overload rats and to explore the mechanism.Methods Aldosterone overload rat myocardial infarction model was obtained by ligating the left anterior descending coronary artery.The pGLV-shRNA was constructed,sequenced and injected into rats via tail vein.Rats were divided into 3 groups:pGLV-shRNA group (n=6),pGLV-shRNA-NC group (n=6,contained a nonsense shRNA) and the sham-operation group (n=6).Cardiac function was measured by cardiac ultrasound.Apoptosis was assessed by transferase (TdT)-mediated biotin-16-dUTP nick-end labelling (TUNEL).The p38 MAPK mRNA expression was analyzed by RT-PCR.The protein expressions of p38 MAPK and caspase-3 were detected by Western blot.Results Compared with the sham-operation group,cardiac systolic function was reduced and myocardial apoptosis index was significantly increased [(31.26 ± 4.45) ％ vs.(15.20 ± 2.18) ％,P ＜ 0.01] in pGLV-shRNA-NC group.The mRNA and protien expressions of p38MAPK and caspase 3 protein expression were significantly increased in pGLV-shRNA-NC group (all P＜0.01).Compared with pGLV-shRNA-NC group,cardiac function was improved,myocardial cell apoptosis index was reduced [(22.35±3.59)％ vs.(31.26±4.45)％,P＜0.05],and the mRNA and protien expressions of p38MAPK and caspase 3 protein expression were decreased in pGLV-shRNA group (all P＜0.05).Conclusions Cardiac dysfunction is associated with p38MAPK-mediated myocardial apoptosis in aldosterone overload MI rats.pGLV-shRNA may inhibit cardiomyocyte apoptosis and improve postMI cardiac function.

Objective To study the early changes of the coagulation system in type 2 diabetic nephropathy.Methods Sixty-two cases of patients with type 2 diabetic nephropathy were divided into two groups:normal albuminuria group ( N-UAlb group,UACR ＜ 30 mg/g,32 cases ),microalbuminuria group ( MUAlb group,UACR:30～300 mg/g,30 cases).Thirty healthy persons constituted a control group (NC group).Fibrinogen( FIB ),antithrombin Ⅲ ( AT-Ⅲ ),protein C ( PC ),protein S ( PS ) were measured by coagulation analyzer,while yon willebrand factor (vWF) and platelet granule membrane protein 140 (GMP-140) were detected by ELISA assay,platelet count (PLT),mean platelet volume(MPV),platelet hematocrit (PCT),platelet distribution width(PDW) by hematology analyzer.Results The level of fibrinogen,GMP-140 and vWF in the M-UAlb group were (4.20 ± 1.53 ) g/L,( 30.03 ± 7.77 ) μg/L,and ( 315.53 ± 47.24 ) ％ respectively,vwhich were significantly higher than those in the N-UAlb group [ ( 3.21 ± 0.89 ) g/L,( 18.22 ± 5.08 ) μg,/L and ( 191.88 ± 57.25 ) ％ respectively ] and the NC group [ ( 2.75 ± 0.53 ) g/L,( 14.26 ± 2.29 ) μg/L and ( 138.12 ± 61.27 ) ％ respectively ] ( F =5.42,10.42,30.44,P ＜ 0.05 or 0.01 ).The fibrinogen,vWF,GMP-140 were positively correlated with UACR ( r =0.313,P ＜ 0.05 ; r =0.620,P ＜ 0.01 ; r =0.680,P ＜ 0.01 ) and PC was negatively correlated with UACR ( r =-0.255,P ＜ 0.05 ).Conclusion Hypercoagulable state in diabetic nephropathy is associated with the high fibrinogen,endothelial dysfunction and platelet activation,and these changes have already emerged in patients without albuminuria.This might mind us that we should strengthen anticoagulant therapy on patients when they are not progressing to albuminuria.

Objective To modify HLA-A2.1 restricted epitope of tyrosinase related protein-2 (TRP-2) (180-188) of human melanoma differentiation antigen and enhance HLA-A2.1 molecule its affinity to. Methods The TRP-2 (180-188) nave epitope was altered with the predominant amino acid for the primary anchor residues and auxiliary anchor residues. Altered peptide ligands (APLs) were scanned by polynomial method, the quantitative motif method and OSAR methods. The analogues their affinity to HLA-A2.1 molecule were assayed. Results Compared with the nave peptide, APLs had stronger affinity to HLA-A2 molecules. Conclusion Our results suggest that altered nave epitope with P1 (Y) and/or P2(L, M) can enhance affinity to HLA-A2 molecule. However, the immunogical efficacy of APLs needs to be identified in studies in vitro and in vivo.

Objective To evaluate the functional outcomes of different limb salvage procedures in patients with bone and soft tissue sarcomas of the shoulder girdle.Methods From July 2001 to July 2008,16 patients with limb salvage for sarcomas of shoulder girdle were respectively analyzed,including 11 males and 5 females with an average age of 38.4 years (range,17-67).Localizations of the tumors were 8 in the scapula(including 4 chondrosarcomas,1 Ewing sarcoma,and 3 metastases),5 soft tissues of the shoulder girdie(including 2 synoviosarcomas,1 fibrosarcoma,1 hemangioperieytoma,and 1 well-differentiated liposarcoma),and 3 proximal humerus (including 1 osteosarcoma and 2 metastases).Twelve patients were treated with classical Tikhoff-Linberg procedures,and 4 with improved procedures.Results The mean surgical time duration was 3 hours.The mean blood loss was 1600 ml.The mean follow-up time was 40 months.Major complications included 2 cases of delayed wound healing,and 1 ulnar nerve injury.One patient had local recurrence and died of pulmonary metastases 18 months after second operation of interscapulothoracal amputation.The patient with fibrosarcoma also died of pulmonary metastases 23 months later.One patient with synoviosarcoma was alive with pulmonary metastases in 9 months.Four of 5 patients with carcinoma metastases died during 11 to 23 months later.The 5-year cumulative survival rate was 34.6%.Functions were preserved in the whole hand and elbow.The MSTS functional score of the patients receiving classical Tikhoff-Linberg procedures was 14.7,while improved Tikhoff-Linberg procedures was 19.5.Conclusion The Tikhoff-Linberg procedure not only provides a wide resection of tumors in the shoulder girdle but also preserve the whole hand and elbow functions.The shoulder function was poor in patients receiving classical Tikhoff-Linberg procedures.

Objective To explore the effect of Occupational Therapy in Group on cognitive function of children with March to cerebral palsy, and to observe the evaluation results of P300 of event-related potentials on cognitive function. Methods 40 children with spastic cere-bral palsy from March to September, 2015, were divided into control group (n=20) and observation group (n=20). Both groups received rou-tine rehabilitation training, and the observation group received Occupational Therapy in Group additionally, for 3 months. They were as-sessed with Wechsler Preschool and Primary Scale of Intelligence (WPPSI) and P300 of event-related potentials before and after treatment. Results The latency of P300 shortened after treatment (t>7.778, P<0.001), and the amplitude of P300 was larger after treatment (t>5.808, P<0.001) in both groups, especially in the observation group (t>0.461, P<0.05). The score of WPPSI increased after treatment in both groups (t>5.944, P<0.001), and was higher in the observation group than in the control group (t=-3.472, P=0.001). Conclusion Occupational Ther-apy in Group could further improve the cognitive function of children with spastic cerebral palsy. P300 can be used as an assessment tool for cognitive function.

Objective To investigate the mechanism of IL-1β in promoting glial scar formation after spinal cord injury.Methods The experimental model of SCI was created by extradural compression of the spinal cord using an aneurysm clip.Rats were randomly divided into model group, sham operation group, IL-1β inhibitor IL-1RA group, IL-1β group and IL-1β+JAK2-STAT3 inhibitor AG490 group, according to different interventions, then were given normal saline, IL-1RA, IL-1β and IL-1β+AG490 every 10 μL respectively, sham group received only laminectomy.The motion function of the hindlimbs of rats was measured by Basso Beattie Bresnahan(BBB) scores and the expression of GFAP, vimentin and p-STAT3 were detected by Western blot technique, immunofluorescence assay and immunohistochemistry technique at corresponding time points(at the 8th, 12th hour, 1st, 3rd, 7th and 14th day after SCI).Results The expression trend of p-STAT3(at the 8th and 12th hour after SCI),GFAP and vimentin(at the 7th and 14th day after SCI)was: the expressions of p-STAT3, GFAP and vimentin in the model group were significantly higher compared with the sham group(P<0.01), the expression of p-STAT3,GFAP andvimentin in the IL-1RA group were significantly lower compared with the model group(P<0.05) whereas significantly higher compared with the sham group(P<0.05);the expressions of p-STAT3, GFAP and vimentin in the IL-1β+AG490 group were significantly lower compared with the model group(P<0.05)whereas significantly higher compared with the sham group(P<0.05), the expressions of p-STAT3, GFAP and vimentin in the IL-1β group were significantly higher compared with the model group(P<0.05).Conclusions IL-1β can improve glial scar formation via JAK2-STAT3 signal.Inhibition of IL-1β or JAK2-STAT3 can reduce glial scar formation and promote functional recovery of spinal nerve.