Aim To study the effect of PLC on rabbit and human platelet actin polymerization, and then to explore the mechanism of PLC anti-aggregation to platelet. Methods Platelets of rabbit and human were treated with PSS, ASA and different doses of PLC respectively and then were extracted by Triton abstraction. The relative concentration of actin of differently treated platelets induced by ADP was determined by SDS-PAGE and spectrophotometre. Results For rabbit platelets were treated with PSS, the relative concentration of actin determined at static state was 1.682?0.319; when the platelets were treated with ASA 668 ?mol?L -1,PLC 5,10,15,20 and 25 U?ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.450?0.562,1.089?0.322,1.727?0.442,1.450?0.324,1.161?0.306, 0.857?0.242 and 0.692?0.187 respectively. Compared with PSS, inhibition rates (%) of ASA 668 ?mol?L -1, PLC 5, 10, 15, 20, 25 U?ml -1 to the relative concentration of actin were 55.55,29.51, 40.82,52.61, 65.02,71.76 respectively.For human platelets were treated with PSS, the relative concentration of actin determined at static state was 1.358?0.376; when the platelets were treated with ASA 668 ?mol?L -1,PLC 5,10,15,20 and 25 U?ml -1, the relative concentration of actin determined at activated state induced by ADP was 2.445?0.750, 1.096?0.344, 1.705?0.507,1.437?0.416, 1.165?0.355, 0.845?0.257 and 0.679?0.198 respectively. Compared with PSS, inhibition rates (%) of ASA 668 ?mol?L -1, PLC 5, 10, 15, 20, 25 U?ml -1 to the relative concentration of actin were 55.17,30.27, 41.23,52.35, 65.44, 72.23 respectively. Conclusion PLC has significant effects on actin polymerization of rabbit and healthy human platelets (P

Objective To investigate the survival and prognostic factors in patients with gastrointestinal stromal tumors(GIST).Methods The histopathological slides from 153 patients with GIST were reviewed. The expression of CDllT,CD34,platelet-derived growth factor receptor alpha (PDGFR-a) and Ki-67 proteins were measured by immunohistochemical staining. The factors thatinvolved in the survival and prognosis were analyzed based on the clinical features and GIST biological behavior ranking.The Kaplan-Meier and Cox model were used to evaluate the effect of variant factors on survival and prognosis.Results The survival rate of 135 patients was 94.1% 76.3% and 65.9% at 1,3 and 5 years,respectively.On univariate analysis survival was predicted by tumor size (χ2= 40.565,P＜0.01),primary tumor location (χ2=13.245,P＜0.01),mitotic count (χ2= 22.626,P＜0.01),risk ranking (χ2=19.186,P＜0.01),necrosis (χ2==28.665,P＜0.01),incomplete resection χ2=2 =29.110,P＜0.01) and Ki-67 index (χ2=15.953,P＜0.01).Multivariate analysis demonstrated that thetumor size ＞ 10 cm,primary tumor location,mitotic count＞ 10/50 HPF,high risk subgroup,tumor necrosis and Ki 67 index ＞ 5 % were poor predictors of survival.Ki-67,tumor size and mitotic count were strong poor predictors of survival.Conclusions Fletcher's biological behavior ranking is a good approach to predict prognosis of GIST patients and has significant clinical value.It's better to combine itwith other factors such as Ki-67 index and primary tumor location in order to provide evidence for therapy.

Objective To identify the key factors triggering medical disputes induced by differences in cognition between doctors and patients in order to control the occurrence and development of medical disputes. Methods From 2002 to 2006,the outpatients,inpatients and medical staff of 3 hospitals in Shanghai(class 3 first level general hospital,n=2;class 3 first level special hospital,n=1) were investigated,and random sampling method was employed for the research analysis. Results There were significant differences between doctors and patients in cognition of influential factors triggering medical disputes such as health law,medical services,rights and obligations. Conclusion The differences in cognition between doctors and patients are important causes for medical disputes,and effective prevention and intervention measures must be taken.

Objective To explore the relationship between polymorphism of miR-499A > G with inci-dence and prognosis of colorectal cancer in Han Chinese. Method 1:1 matched case control study was used to collect samples,and the patients were followed up. The genotypes distribution miR-499A > G were detected by TaqMan probe,and logistic regression model and Cox proportional hazard regression model were used to analysis the relationship between miR-499A>G and incidence and prognosis of colorectal cancer. Results 394 cases and 394 controls were recruited,while miR-499A>G did not increase the risk of colorectal cancer(OR=1.22,95%CI:0.95-1.57)under additive model analyzed by multi-logistic regression model. The means of survival time of AA,AG,GG carriers were 54.19 ± 2.02,44.16 ± 2.59,32.76 ± 5.36 months and there were significant differentia-tion between them in log Rank test(F=11.24,P=0.004). Cox proportional hazard regression model showed that miR-499A > G was significant associated with adverse prognosis of colorectal cancer(HR=1.26,95%CI:1.02-1.57). Conclusion miR-499 A > G has no association with the incidence of colorectal cancer ,but it affects the prognosis of colorectal cancer in Han population.

Objective To investigate the change of serum Nogo-A protein in patients with acute closed brain injury, and explore its relationship with the severity of neuronal damage and prognosis. Methods Thirty-one patients with acute closed brain injury were enrolled. Venous blood samples (2 mL) were obtained 1, 3 and 5 d after injury. Serum concentrations of Nogo-A protein were determined by ELISA. Patients were divided into mild (n =7), moderate (n = 10) and severe (n = 14) injury groups according to Glasgow coma score (GCS), and were divided into favorable prognosis (n = 23) and poor prognosis (n = 8) groups according to Glasgow outcome score (GOS). Another 20 healthy adults were served as controls. Results The mass concentrations of serum Nogo-A protein in mild, moderate and severe injury groups 1, 3, 5 d after injury were significantly higher than those in control group (P < 0.01), and the mass concentrations of serum Nogo-A protein in moderate and severe injury groups 1, 3, 5 d after injury were significantly higher than those in mild injury group (P <0.05, P <0.01). The mass concentrations of serum Nogo-A protein 1, 3, 5 d after injury were significantly higher in poor prognosis group than those in favourable prognosis group (P < 0.01). Conclusion Serum Nogo-A protein level significantly increases after brain injury, and is related to the degree of injury and prognosis.

AIM Aim to study the effect of PLC on human platelet cytoplasmic free calcium concentration, and then to explore the mechanism of PLC anti aggregation to platelet. METHODS Washed platelet of human was marked with fluorescence indicator Furu 2/AM, and then was treated with PSS, ASA and different dose of PLC respectively. Cytoplasmic free [Ca 2+ ] i of differently treated platelet at static state and activated sate induced by ADP was determined by double beam fluorescence spectrophotometric method. RESULTS Healthy human platelet was treated with PSS, ASA 334 ?mol?L -1 , 2 5, 3 75, 5, 10 and 20U PLC?ml -1 . Cytoplasmic [Ca 2+ ] i determined at static state was 152 55?15 07, 131 63?15 58, 140 27?12 03, 139 48?1 73, 121 11?9 58, 116 62?15 96 and 107 20?17 07 respectively. [Ca 2+ ] i determined at activated sate induced by ADP was 902 62?94 74, 687 99?62 86, 810 99?72 37, 701 73?21 37, 429 67?71 59, 342 82?44 86 and 263 27?25 46 respectively. Compared with PSS, inhibition rates ( % ) of ASA 334 ?mol?L -1 , 2 5, 3 75, 5,10, 20 U PLC?ml -1 to the increment of activated platelet cytoplasmic [Ca 2+ ] i were 25 83, 10 58, 25 04, 58 86, 69 84, 79 19 respectively. At the same condition, inhibition rate ( % ) of 10 U PLC?ml -1 to the increment of five hypertensive activated platelet cytoplasmic [Ca 2+ ] i were 74 25, 73 48, 61 32, 82 87,70 60 respectively. CONCLUSION PLC has significant effects on static cytoplasmic[Ca 2+ ] i of healthy and hypertensive human platelet ( P

BACKGROUND: Magnetic resonance diffusion tensor imaging (MRDTI) may non-wounded detect damage of fiber in white matter and becomes an effectively way to evaluate upper motor neuron(UMN) impairments.OBJECTIVE: To investigate the clinical significance of MRDTI on amyotrophic lateral selerosis(ALS).DESIGN: Case contrast observation.SETTING: Department of Neurology, Renmin Hospital of Wuhan University.PARTICIPANTS: Twenty ALS patients were selected from Department of Neurology, Renmin Hospital of Wuhan University from April to December 2005. There were 11 males and 9 females, and their ages ranged from 33 to 73 years with the mean age of (51±10) years. All subjects met the diagnostic criteria of ALS set by World Neurology League.Other 15 healthy subjects were collected as control group. There were 8 males and 7 females, and their ages ranged from 31 to 73 years with mean age of (50±11) years. All subjects provided the confirm consent.METHODS: Based on level of upper and lower motor neuron impairments, ALS patients were divided into UMN impairment group (n =16) and lower motor neuron group (n =4). Functional scores of ALS, illness developing velocity and pyramidal sign scores were performed, respectively. All subjects were scanned with DTI at axial view. Regions of interest [subcortical white matter of precentral gyrul and postcentral gyrul (Pre-CG/Post-CG), centrum semiovale and frontal white matter (CS/FWM), peripheral lateral cerebral ventricle, posterior limb of internal capsule (PIC), cerebral peduncle (CP), genu corpus callosum and splenium corpus callosum (GCC/SCC) and dorsal thalamus (DT)] were selected to measure fractional anisotropy (FA) and apparent diffusion coefficient(ADC).MAIN OUTCOME MEASURES: Correlations among FA, ADC, functional score of ALS, illness developing velocity and pyramidal sign scores.RESULTS: Twenty patients and 15 subjects in the control group were involved in the final analysis. ① FA was reduced and ADC increased in the posterior limb of the internal capsule in patients with UMN signs compared to healthy volunteers (t =3.452, 2.670; P ＜ 0.01, 0.05). Nonparametric tests revealed that there was a trend toward reduced FA in the posterior limb of the internal capsule in B group compared to controls (U =11, P =0.057). ② In UMN impairment group, FA in the posterior limb of the internal capsule was positively correlated with the ALS rating scale (r =0.577, P ＜0.05) and negatively correlated with pyramidalsign scores (r = -0.789, P ＜ 0.01 ),CONCLUSION : The impairment of pyramidal tracts can be noninvasively evaluated by diffusion tensor MR in vivo, thus providing useful information in diagnosing and further understanding MND.

Objective To investigate the feasibility of laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation for rectal cancer.Methods The clinical data of 34 patients with rectal cancer who had been admitted to OUr hospital from January 2007 to December 2007 were retrospectively analyzed.All patients underwent laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation.The operation time,lymph node dissection,postoperative complications were assessed.The erectile function of male patients and the short-term outcome of the operation were evaluated.Results All patients were successfully operated on,and no conversion Was required.The mean operation time,blood loss,number of lymph node dissected,time of catheterization,mearl volume of residual urine and CEA were(265+46)minutes,(123±27)ml and 19±3,(5.5±1.6)days,(22.5±7.8)ml and(8.0±4.6)U/L,respectively.The incidence of postoperative complications and male sexual dysfunction were 9%(3/34)and 14%(3/21),respectively.No local recurrence or distal metastasis Was observed at the end of August 2008.Conclusions Laparoscopic D3 lymphadenectomy combined with pelvic autonomic nerve preservation for rectal cancer is relatively safe and feasible.

The objectives of this study were to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudotype virus on SiHa cytobiology behavior by cutting the HPV16 E6 gene selectively and to explore the role of this system in the treatment of cervical cancer.After designing specific gRNA sequences targeting HPV 16 E6,generating hCas9-EGFP and E6-gRNA-RFP plasmids,and preparing the pseudovirus of HPV16 carrying E6-gRNA and Cas9 plasmids,we determined the titer of the pseudotype virus using the TCID50 method.We obtained the pseudotype virus of HPV16 carrying E6-gRNA and Cas9 plasmids to transfect cervical cancer SiHa cells.Experimental subjects were divided into control group,empty virus group,E6-gRNA transfected group,Cas9 transfected group and Cas9+E6-gRNA transfected group.The molecular size of the cutting sequence was detected using the T7E1 enzyme digestion method and agarose gel electrophoresis,and the cleavage function of CRISPR/Cas9 on the E6 gene was determined at the same time.RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of E6 in all the groups;the Transwell cell migration assay was performed to detect the cell migration ability and metastasis in all groups.Heterotopic transplantation tumors were incorporated into mice and were used to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudovirus on the tumorigenic ability of SiHa cells by selectively cutting HPV16 E6.The HPV16 pseudotype virus carrying E6-gRNA and Cas9 plasmids could successfully infect SiHa cells,and there were two cutting zones in the Cas9+E6-gRNA transfected group.However,the empty virus group,E6-gRNA transfected group and Cas9 transfected group had no corresponding zone.Compared with those in the control group,the empty virus group,E6-gRNA transfected group and Cas9 transfected group,the mRNA and protein expression levels of E6 in SiHa cells were downregulated in the Cas9+E6-gRNA transfected group (P＜0.01).In addition,the proliferation and migration abilities of SiHa cells were significantly inhibited (P＜0.01).There were no significant differences among the other groups.In contrast to the control group,the HPV pseudotype virus carrying E6-gRNA and Cas9 plasmids could significantly delay the growth of tumor cells of the ectopic tumor transplantation model (P＜0.01).The CRISPR/Cas9 system mediated by the HPV pseudotype virus to knockout E6 gene expression exhibited a clear inhibitory effect on the biological function of SiHa cells,which indicated that knocking out the E6 gene using the CRISPR/Cas9 system mediated by the HPV pseudotype virus had a potential effect of eliminating HPV infection and inhibiting the growth of HPV-related tumors.Taken together,these findings provide insight into a new treatment strategy for the prevention and treatment of hr-HPV infected disease,particularly in HPV-related tumors.

Objective To study the expression of VEGF-C and it's relationship with pathologic characteristics in rectal cancer.Method In this study 82 patients of mid to low rectal cancer underwent radieal resection from December 2007 to August 2008.Tumor tissues and lymph nodes were studied by immunohistochemical staining for VEGF-C expression in the tumor tissue and CK20 expression in D3 lymph nodes.Results were compared with the pathological results of mesenterium lymph nodes to explain the relationships between VEGF-C expressions and clinical pathologic characteristics.Data were analyzed with Chi square test.Result As for the expression of VEGF-C.significant differences were found between tumor stages(X~2=8.529,P<0.05)and between positive and negative lymph node metastasis(X~2=4.712,P<0.05).but there was no difference between that of mesenterium lymph node metastasis and D3 lymph node micrometastasis(X~2=0.017,P>0.05). Conclmion In rectal cancer,VEGF-C expression is correlated with tumor stage,type,metastasis and micrometastasis of lymph node.