Objective To study the predicting value of four different scoring systems such as the acute physiology and chronichealth evaluation Ⅱ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, quick SOFA (qSOFA) score and systemic inflammatory response syndrome (SIRS) score for the prognosis of septic patients. Methods A retrospective analysis were conducted. Septic patients in intensive care unit (ICU) of the First People's Hospital of Chenzhou form July 1st, 2012 to June 30th, 2016 were enrolled.Patients were divided into survival group and death group according to 28-day outcome. The difference of clinic data, the worst clinical index value within 24 hours, whether mechanical ventilation performed on first day, length of stay in ICU, APACHE Ⅱ score, SOFA score, qSOFA score and SIRS score were compared between the two groups. The significant different factors of sepsis outcome in univariate analysis were analyzed by multiple logistic regression, and the ability of four scoring systems was tested by receiver operating characteristic (ROC) curve.Results 311 patients were enrolled in this study (221 survivals, 90 deaths, 28-day mortality rate 28.9%). Univariate analysis showed age, mechanical ventilation ratio, urine output, length of stay in ICU and the fastest heart beat rate (HR), the lowest systolic blood pressure (SBP), the lowest mean arterial pressure (MAP), HCO3-, minimum arterial blood oxygen partial pressure (PaO2), minimum oxygenation index (PaO2/FiO2), the maximum fraction of inspired oxygen (FiO2), Na+, the highest concentration of blood urea nitrogen (BUN), the highest concentration of serum creatinine (SCr), minimum concentration of plasma albumin (Alb), Glasgow coma score (GCS) score, APACHE Ⅱ score, SOFA score, qSOFA score, within 24 hours after diagnosis were significantly different between two groups (allP < 0.05). Multiple logistic regression showed age [odds ratio (OR) = 1.388, 95% confidence interval (95%CI) = 1.074-1.794,P = 0.012], PaO2/FiO2 (OR = 0.459, 95%CI = 0.259-0.812,P = 0.007), concentration of plasma Alb (OR = 0.523, 95%CI = 0.303-0.903,P = 0.020), GCS score (OR = 0.541, 95%CI = 0.303-0.967,P = 0.038) and SOFA scores (OR = 3.189, 95%CI = 1.813-5.610,P = 0.000) were independent risk factors for sepsis outcome. ROC curve test showed the APACHE Ⅱ score, SOFA score and qSOFA score had the ability to predict the outcome in critical ill patients with sepsis, the SOFA score of the most powerful, the area under the ROC curve (AUC) was 0.700,when the cut-off value was 7.5 points, the sensitivity was 73.3% and specificity was 58.8%.Conclusions APACHE Ⅱ score, SOFA score and qSOFA score have the predictive properties for septic patients. SOFA score is an independent prognostic risk factor of sepsis, while qSOFA score can be widely used in clinical practice as the advantage of quick evaluating.

Objective:To screen the binding peptide against adenovirus type 7(Ad7) and evaluate the relevance with the ade-novirus receptor .Methods:Binding peptide against Ad 7 was screened by panning the phage display 12 peptides library .The antibody against the selected peptide was prepared and was used to study the binding to the membrane by immunofluorescence technique .Re-sults:Using Ad7 as the target protein , GTS09 peptide was selected from the phage display 12 peptides library by biopanning .GTS09-phage complex could significantly bind Ad 7, with the affinity constant up to 1.93 ×1010 L/mol;at the same time, immunofluorescence showed that antibody of GTS09 could specifically bind to membrane of 293 cell.Conclusion: Antibody against GTS09 peptide could specifically bind to membrane of 293 cell,which shows that the peptide may presumably have homology with the cell receptors of Ad 7.

Objective To clone and express the hexon protein of three prevalent human adenovi -rus strains causing respiratory disease and analyze the antigenic characteristics of the recombinant proteins . Methods The full length genes encoding hexon protein of human adenovirus serotype 3(HAdV3), serotype 4(HAdV4) and serotype 7(HAdV7) were cloned by PCR and sequenced , respectively.The alignment anal-ysis was performed by using hexon gene sequences from GenBank .The major antigenic regions of hexon pro-tein of the three serotypes were expressed in E.coli and purified.The antigenicity, immunogenicity and cross reactivity of the recombinant proteins were determined by ELISA and Western blot assay .Results The full length gene sequence encoding hexon protein of human adenovirus serotype 4 was firstly reported in China , which showed more than 99%homology in both nucleotide and amino acid sequences with the human adeno-virus type 4 NHRC3 strain.The partial hexon protein sequence of HAdV 3, HAdV4 and HAdV7 containing all of the 7 hyperviriable regions ( HVRs) were expressed in E.coli, respectively .The purified recombinant proteins could be recognized by antiserum of the three serotypes of adenovirus .The antiserum samples against the three recombinant proteins could cross-react with particles of the three serotypes of adenovirus . The possible type-and species-specific epitopes were predicted .Conclusion The major antigenic regions of hexon protein of the three serotypes were successfully expressed .The purified recombinant proteins contai-ning both intertypes and type-specific epitopes showed a strong immunogenicity .

Objective To investigate the effect of low molecular weight heparin (LMWH) on the expression of endothelial protein C receptor (EPCR) and protease activated receptor 1 (PAR1) in abdominal vascular endothelial cells (VECs) of septic rats. Methods VECs were cultured by tissue-sticking method, and the purity was determined with flow cytometry (FCM). VECs were randomly divided into three groups: control group, septic group (LPS 1 μg/ml) and LMWH group (LPS 1 μg/ml+LMWH 5 μg/ml). The VECs were collected at 1st, 3rd, 5th days after stimulated. The expression of EPCR and PAR1 were assessed by FCM. Results The expression of EPCR and PAR1 of septic group decreased significantly compared with control group at each time point (P＜0.05 or P＜0.01), and the expression decreased most obviously on day 5 (26.53±7.21 vs 39.26±2.62,q＝6.45,P＜0.01;53.21±15.10 vs 86.54±11.34,q＝6.94,P＜0.01). In LMWH group, the levels of EPCR and PAR1 expression were higher than setpic group at each time point (P＜0.05). Compared to control group, the expression of EPCR had a significantly decrease on day 1 (40.86±1.63 vs 45.41±2.82,q＝3.51,P＜0.05), which had no significantly different on day 3 and 5 (41.20±3.32 vs 42.83±2.66,P＞0.05;39.23±3.33 vs 39.26±2.62,P＞0.05), and the expression of PAR1 were not significantly decrease compared with control group at each time point (P＞0.05). Conclusions LMWH could improve the inhibition status and the expression of EPCR and PAR1 on VECs in septic rats.

Objective To observe the effectiveness and safety of oral anticoagulants and antithrombotic therapy in elderly patients with atrial fibrillation. Methods Patients were divided into anticoagulant group (warfarin) and antithrombotie group (aspirin or clopidogrel) based on the initial treatment. The prothrombin time (PT), activated clotting time (ACT), international normalized ratio (INR), activated partial thromboplastin time (APTT), fihrinogen (FIB), thrombin time (TT), coagulation factor Ⅱ,Ⅴ,Ⅶ,Ⅷ,Ⅸ, and Ⅹ,fibrin degradation product (FDP) and D-dimer were tested at baseline and after therapy in both groups. Results The average treatment period was 44.2±37.5 months in antithrombotic group and 39.0±61.5 months in anticoagulant group. There were six cases of isehemic stroke, one acute artery embolism in right lower limb and three gastrointestinal bleeding in antithrombotic group, while two gastrointestinal bleeding and two fatal hemorrhagic stroke in anticoagulant group. The results of PT, ACT, INR, APTT, FIB, TT, coagulation factor Ⅱ,Ⅴ ,Ⅶ, Ⅷ,Ⅸ,Ⅹ,FDP and D-dimer had no significant differences compared with the baseline in antithrombotic group. However, there were significant increase in PT and INR [(8.4±7.5)s and (0. 93±0. 83)s, both P<0. 05)], and significant decrease in ACT, coagulation factor Ⅱ,Ⅶ, Ⅸ and Ⅹ (all P<0. 05) in anticoagulant group. Conclusions Anticoagulant therapy may he effective in prevention of ischemic stroke in elderly patients with atrial fibrillation. However, it may slightly increase the hemorrhage incidence. The overall adverse events were not significantly reduced.

Objective To examine the relationship between premature ventricular contraction(PVC) and microvascular leakage in the myocardium due to microvascular contrast echocardiography(MCE). Methods Thirty eight rats were randomized into 5 groups: control, ultrasound exposure only, contrast agent only, real time MCE and trigger imaging MCE. Anesthetized rats with tail vein catheter were imaged in a 37℃ water bath at left ventricular parasternal short axis view. Optison was injected at a dosage of 5 ml/kg. Frequency was 1.7 MHz, MI= 1.7 and the depth of image at 10 cm. Results PVC was detected in all ultrasound exposure groups, together with petechial hemorrhages and Evans Blue leakage in the scan band. The triggered imaging showed worse effects than real time imaging. No PVC or microvascular leakage was noticed in controls or sham exposed rats. Conclusions Induction of PVC during contrast added echocardiography is associated with microvascular leakage.

BACKGROUND: Pathological change of synovium in rheumatoid arthritis (RA) has the characteristic of tumor-like growth, it appears thickening of the synovium tissue and the formatiom of pannus, which generate periarticular erosion and destruction. Multiplicate cell factors and growth factors participate in the development course of tumor-like lesion of synovium, and the tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF)play important roles in the development of RA and the formation of pannus.OBJECTIVE: To observe the contents of plasma TNF-α of collagen-induced arthritis and the expression change of VEGF of synovium at different time point, and investigate the effect and correlation of TNF-α and VEGF in the pathogenesis of RA.DESIGN: Randomized grouping experiment taking animals as subjects.SETTING: Institute integrated traditional and western medicine of Xiangya Hospital of Central South University.MATERIALS: The experiment was finished from July to November 2003 in the laboratory of institute integrated traditional and western medicine.Forty SD rats aged 45-50 days were selected, the rats were randomly divided into the normal control group (n=10) and collagen-induced arthritis model group(n=30).METHODS: 10 mg Cattle collagen type Ⅱ and 5 ml full Freund adjuvant were grinded together, 100 μL of which were intradermally injected at the root of tails of rats, collagen-induced arthritis model of rats were re-immunized with the above-mentioned methòd and dosage after 21 days. The accumulated points of arthritis index was evaluated based on degree and extent of flare and the condition of tumefaction and deformation, the higher the accumulated points of arthritis index were, the more serious the arthritis symptom was. The blood was obtained by decapitation after 25 days in normal control group, and in the collagen-induced arthritis model group the blood was obtained by decapitation 25,30,35,40,45 days after immunization (model establishing), the contents of plasma TNF-α of collagen-induced arthritis rats at different time point was detected by radio-immune assay, the level of expression of VEGF in synovium tissue were detected with immunohistochemical method simultaneously, the correlation of invasion time and the neovascularization of synovium. The accumulated points of arthritis index . TNF-α and VEGF was observed. The correlation of TNF-α and VEGF was analyzed with the linear correlation analysis, the correlation of the accumulated points of arthritis index and TNF-α. VEGF was analyzed with the rank correlation analysis.MAIN OUTCOME MEASURES: The correlation of invasion time of collagen-induced arthritis with the accumulated points of arthritis index, with the plasma content of TNF-α and the expression of VEGF, the correlation analysis of the plasma content of TNF-α of collagen- induced arthritis rats with the expression of VEGF in synovium.RESULTS: Forty rats attended the experiment, all of them entered the final analysis. the neovascularization of synovium was increased, synovium was thickening, the accumulated points of arthritis index was gradually increased, and the contents of TNF-α and level of VEGF were increased accordingly with the process of the invasion time of the collagen-induced arthritis rats; Its accumulated points of arthritis index had the positive correlation with the level of expression of VEGF (r=0.535 ,P ＜ 0.05)and had the correlated increasing tendency with the contents of TNF-α, but there was no significant difference(r=0.371 ,P ＞ 0.05 ). the plasma contents of TNF-α had the positive correlation with the level of expressions of VEGF (r=0.893,P ＜ 0.01 ).CONCLUSION: The TNF-α and VEGF have an important effect on the inflammatory reaction of RA and Cytokine network of neovascularization of synovium, they are possibly mutually influenced and promoted and have the effect of mediating the malignant network circulation; They are the key factors among multiplicate ones which mediate the generation and development of RA, bone erosion and Mutilation.

Objective To explore the neuropsychological features of cognitive impairment induced by chemotherapy for breast cancer patients.Methods A neuropsychology battery was applied in this study.Seventy six breast cancer patients were enrolled in the test and classified as chemotherapy treatment patients(CT,n =38),and non-chemotherapy treatment patients(non-CT,n =38).Forty normal female people were also evaluated as healthy control(HC).Results Compared with HC and non-CT groups,the correct number of backward(CT:4.42±1.11,non-CT:5.18 ± 1.16,HC:5.13 ± 1.22),delayed recall (CT:8.55 ± 1.75,non-CT:9.58 ± 1.50,HC:10.13 ± 1.92) and recognition (CT:7.68 ± 1.90,non-CT:8.97 ± 1.62,HC:9.08 ± 2.09) were low in the CT group (P ＜ 0.01).The reaction time of Stroop test B (CT:(21.54 ± 5.02) s,non-CT:(19.37 ± 4.26) s,HC:(18.82 ± 3.05) s),Stroop test C (CT:(34.85 ± 8.46) s,non-CT:(31.02 ± 7.38) s,HC:(30.61 ± 7.83) s) and TMT test B(CT:(102.79± 11.90)s,non-CT:(96.22 ± 12.07) s,HC:(97.21 ± 11.64)s) were long in the CT group (P ＜ 0.05).There were no significant differences in the Forward,Immediate Recall,Stroop test A,TMT test A and VFT among three groups(P＞ 0.05).Conclusion Breast cancer patients with chemotherapy treatment have cognitive impairment in the domains of memory,attention and executive functions.

Objective To construct a hexon-chimeric human adenovirus type 3 ( HAd3 ) vector expressing two neutralizing epitopes of hepatitis B surface antigen preS 1 (HBsAg-preS1) and to analyze the antigenicity of the chimeric epitopes .Methods Two neutralizing epitopes of HBsAg-preS1 including KR359 and KR127 were inserted into hypervariable region 1 ( HVR1) and hypervariable region 2 ( HVR2) of HAd3 hexon .Chimeric hexon gene encoding the two epitopes was amplified by overlap PCR and then subcloned in -to shuttle plasmid pBR322-L/R containing the homologous recombination regions .The digested shuttle plas-mid containing chimeric hexon gene was co-transfected into Escherichia coli BJ5183 cells together with back-bone plasmid pBRAdΔE3GFP to construct pBRAdΔE3GFP-preS1 vector.Then pBRAdΔE3GFP-preS1 vector was digested with AsiSⅠand transfected into AD293 cells to construct recombinant virus (rAD3E-preS1). CsCl gradient centrifugation was used for purification .Glutathione S-transferase ( GST ) fusion protein KR359KR127 ( GST-KR359KR127 ) was expressed in Escherichia coli BL21 by using expression vector pGEX-4T3.Female BALB/c mice at age 6-8 weeks were intraperitoneally injected with 1010 virus particles or 80 μg GST fusion protein .Serum samples were collected to analyze the antigenicity of two epitopes by ELISA and Western blot .Results ELISA showed that KR 359 and KR127 were successfully exposed on viral sur-faces by using hexon-chimeric HAd3 vector .The induced polyclonal antibodies in serum samples could rec-ognize GST fusion protein and native HBsAg from patients infected with hepatitis B virus .Conclusion The antigen capsid-incorporation strategy could be used to display epitopes on viral surface .Enhanced antigen-specific responses could be achieved through inserting multiple foreign epitopes into hexon HVRs .This study provided evidence for further application of hexon -chimeric human adenovirus type 3 vector in the developmentof vaccine, especially for the development of multivalent hepatitis B vaccine .

Objective To prepare recombinant human adenovirus type 3 expressing Norovirus cap-sid protein gene(Noro-orf2). Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of in-fantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV-Nor was linearized with EeoR Ⅴ and Not Ⅰ, and transformed into E. coil BJ5183 with lined edenovirus ge-nomic DNA pLasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfeeted into Hep-2 cells with LipofectAMINETM 2000 to package recombi-nant adenovirus particles. Results Noro-orf2 was successfully inserted into the shuttle vector. The recombi-nant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and puri-fied. Norovirus eapsid protein gene expression was confirmed in Hep-2 cells by immunecytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus eapsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.