BACKGROUND:Erythropoietin and progenitor cel transplantation both have therapeutic effects on lower extremity arterial occlusive disease.
OBJECTIVE:To investigate the erythropoietin modification effect and magnetic resonance imaging feasibility of superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cel s in vitro.
METHODS:Rat bone marrow-derived endothelial progenitor cel s at logarithmic growth phase were randomized into four groups:endothelial progenitor cel group, SPIO labeled transfection group (pcDNA3-EPO transfection fol owed by SPIO labeling), SPIO labeled empty vector group (empty plasmid transfection fol owed by SPIO labeling), and SPIO labeling group (only SPIO labeling). 4.7T MRI was used to observe SPIO-labeled endothelial progenitor cel s. Cel proliferation, cel cycle distribution, and expression of erythropoietin protein in the four groups were measured.
RESULTS AND CONCLUSION:MRI findings showed with the increasing cel number, gradual y lowered signal intensity on T1-weighted imaging (T1WI), T2WI and T2*WI was seen, and the reduction in the signal intensity was the maximum on T2*WI sequence and the minimum on T1WI sequence. For T1WI, T2WI and T2*WI sequences, the minimum number of cel s was 2×104, 1×104 and 0.5×104, respectively. Cel proliferation and cel cycle distribution showed no significant difference among three SPIO labeling groups. In addition, the expression of erythropoietin protein was only found in the SPIO-labeled transfection group. These findings showed that under SPIO labeling, erythropoietin gene-modified endothelial progenitor cel s show no changes in cel proliferation and cel cycle, and the 4.7T MR is capable of imaging SPIO-labeled erythropoietin gene-modified endothelial progenitor cel s in vitro.

ObjectiveTo study the way for establishing prediction model of acute pancreatitis with secondary multiple organs dysfunction syndrome (MODS) in the early stage.MethodsThe linical data were collected from 113 hospitalized patients with acute pancreatitis admitted from October 2008 to October 2011.Some biomarkers within 48 h after the onset of acute pancreatitis were statistically analyzed by univariate and multivariate binary logistic regression analysis in order to establish the prediction model evaluated by the ROC curve. ResultsThere were statistically significant differences in lactate dehydrogenase ( LDH ),serum creatinine ( CR),serum albumin ( P ＜ 0.05 ) identified by univariate regression analysis.These independent risk factors in the regression model were:ln (P/1 -P) =2.243 +0.002 × LDH + 0.014 × CR - 0.186 × ABL,likelihood ratio test ( x2 =66.46,P =0.000) ; in the ROC curve analysis:AUC 0.927 (95％ CI:0.875 -0.980),the best cut- off value of the model predicting probability was 0. 177 (sensitivity 82. 14％,specificity 85.88％,correctly classified 84.96％ ).ConclusionsEfficient prediction model could be set up by the logistic regression analyzing the early risk factors in multiple organ dysfunction syndrome following acute pancreatitis.

Objective To explore the relationship among aggression of patients,doctor-patient trust and perceived social support,and to provide a new prospective of improving doctor-patient relationship.Methods Data of Chinese buss-perry aggression scale,wake forest physician trust scale and perceived social support were collected from a sample of 338 patients in Guangzhou and analyzed with structural equation modeling.Results ①Median (interquartile range) of aggression,doctor-patient trust and perceived social support were 54(18),32(8) and 62(15) respectively.②Total score of aggression(Z=-2.37,P=0.02) and the dimension score of physical-aggression(Z=-4.07,P＜0.01) between different gender were statistically significant,and the scores of male were higher than female.③According to relevant result,aggression of patients was significantly negatively correlated with both doctor-patient trust (r=-0.13,P＜0.05)and perceived social support(r=-0.14,P＜0.01).Doctor-patient trust was significantly positively correlated with perceived social support(r=0.36,P＜0.01) ④Perceived social support fully mediate the relationship between aggression of patients and doctor-patient trust(GFI =0.989,NFI =0.989,IFI =0.998,CFI =0.998,RMSEA =0.026,x2/df=1.229).Conclusion Perceived social support is a protective factor of aggression of patients.Improving the perceived social support of patients can promote the doctor-patient trust.

Objective To investigate the predictive value of serum uric acid for patients with traumatic brain injury.Methods A total of 330 patients with traumatic brain injury (Glasgow Coma Scale score,GCS:3-14) admitted to the First Affiliated Hospital of Soochow University between November 2010 and October 2012 were enrolled.They were divided into a survival group (GOS:2-5) and a non-survival group (GOS:1).The levels of serum uric acid were measured from venous blood in the morning of the second day after admission.Clinical data were analyzed by logistic regression model,spearman correlation,and ROC curve analysis.Results Spearman correlation analysis showed that serum uric acid was significantly correlated with GCS (r =-0.270 1,P =0.000) and GOS (r =-0.251 2,P =0.000).Age,GCS,pupil reaction and serum uric acid were determined as independent predictors for death by logistic model.The adjusted OR of uric acid was 1.0048,(95％ CI:1.001 9-1.007 6,P =0.001).The area under the ROC curve was 0.718,(95％ CI:0.666-0.766),the optimal cut-off value determined by the Youden index was 304 μmol/L (sensitivity:60.24％,specificity:78.14％,correctly classified:73.64％).In the core model (Age + GCS + Pupil reaction),theR2 value was 0.476 4.With uric acid added into,the predictive power of the model increased to R2 =0.510 5 (7.2％ increased).Conclusions The level of serum uric acid is significantly correlated with the severity of TBI and could be used as an independent predictor for death.

BACKROUND:Corneal hemangiogenesis occurs in 40%-60%patients after keratoplasty.Blood vessel is one of the high risk factors for corneal immunological rejection.To inhibit corneal hemangiogenesis would prolong the survival time of the grafts and promote the successful rate of the keratoplasty.OBJECTIVE:To explore the inhibitive effects of doxycycline on corneal angiogenesis after keratoplasty.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the State Key Laboratory of Ophthalmology(No.2006DA105054),Zhongshan Ophthalmic Center,Sun Yat-sen University from March to August 2007.MATERIALS:A total of 48 healthy dean Sprague Dawley rats served as recipients(right eye)and 24 Wistar rats as donors(both eyes).CD31-PEfluorescent antibody was obtained from Sigma,USA.Sandwich enzyme-linked immunosorbent assay(ELISA)kit for vascular endothelial growth factor(VEGF)was brought from RapidBio,USA.METHODS:Corneal allogenic transplantation models were established in rats.Recipients were equally and randomly divided into 2 groups:saline control group and doxycycline group.Twenty minutes prior to surgery,mydriasis was performed using 1%atropine,with a diameter of 2.75 mm of implant and 2.5 mm of implant bed.In the saline control group,conjunctiva of the right eye received saline,three times a day,following surgery.In the doxycycline group,conjunctiva of the right eye received 1%doxycycline,three times a day,till 30 days following surgery.MAIN OUTCOME MEASURES:The following parameters were measured:corneal angiogenesis using immunofluorescence,expression of VEGF protein by using ELISA.RESULTS:Compared with the survival time of saline control group[(9.67±2.73)days],the mean survival time of doxycycline group[(20.67±3.01)days]was significantly prolonged(P＜0.01).The mean percentages of neovascularized corneal area in the saline control group were(4.00±1.00)%,(14.33±4.04)%,(31.33±3.51)%at 3,7 and 14 days after keratoplasty,respectively.The mean percentages of neovascularized corneal area in the doxycycline group were(1.67±1.15)%,(4.67±1.53)%,(18.33±1.53)%at the same time point respectively.Compared with the saline control group,the mean percentages of neovascularized corneal area of the doxycycline group was significantly reduced at 7 and 14 days after keratoplasty(P ＜0.05).The expression of VEGF in the saline control group was(541.00±75.44)pg/mg,(960.00±90.14)pg/mg,(976.00±130.41)pg/mg at 3,7 and 14 days after keratoplasty,respectively,while expression of VEGF in the doxycycline group was(115.33±9.29)pg/mg,(239.00±41.62)pg/mg,(361.00±65.20)pg/mg,respectively.The difference of VEGF expression at all time points between the two groups was significant (P＜0.05).CONCLUSION:Doxycycline has a significant effect in inhibiting corneal angiogenesis and prolonging survival time of implants after keratoplasty.

BACKGROUND: Cervical lymph nodes are draining region of cornea. It is believed that aqueous fluid goes through a minor pathway named uveoscleral drainage, which will allow passage of antigen-presenting cells (APC) directly to the draining lymph nodes and induce allograft rejection after keratoplasty.OBJECTIVE: To explore the inhibitory effects of cervical lymphadenectomy in alkali induced high-risk corneal transplantation.DESIGN: A randomized controlled animal experiment.SETTING: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University.MATERIALS: The experiment was performed in the State Key Laboratory of Ophthalmology (No. 2006DA105054), Zhongshan Ophthalmic Center, Sun Yat-sen University from May 2005 to February 2007. 144 male animals (1-2 months old) including 104 SD rats and 40 Wistar rats were provided by the animal experimental center of Sun Yat-sen University. Sandwich enzyme-linked immunosorbent assay (ELISA) kits for interleukin-2 (IL-2) and interferon-γ (IFN-γ) were brought from BioSource International company (USA). The animal treatment in the experiment was accorded with the statement in Association for Research in Vision and Ophthalmology (ARVO) for animals.METHODS: With the SD rats as recipients, and Wistar rats as donors, all rats were subjected to corneal allografting. The recipient rats were randomly divided into 4 groups (n=20): group A (control group) which underwent corneal transplantation; group B which was subjected to bilateral cervical lymphadenectomy; group C, corneal transplantation 21 days after the alkali burn injury; group D, cervical lymphadenectomy following group C. The immune rejection of grafts was evaluated by detecting the expression of IFN-γ and IL-2 using ELISA. The time when allograft rejection occurred was recorded and mean survival time (MST) was compared among the groups. The development of corneal inflammation and new vessels was examined by slit lamp microscope and histopathological examination.MAIN OUTCOME MEASURES: ①The development of corneal inflammation after corneal alkaline burns. ②MST of rats in each group following transplant. ③The expression of IL-2 and IFN-γ in grafts of each group. RESULTS: ①Normal rat cornea was transparent without inflammation or neovascularization. There were many inflammatory cells invading to stroma on day 3 after burn. Then, the inflammation of cornea resolved gradually 3 weeks after the burn, but corneal neovascularization reached the peak at that time. Corneal blood vessels regressed completely at the end of 8 weeks after the burn. ②The MST of group A, B, C, and D was (10.40±1.14), (46.30±9.46), (7.00±1.58), and (15.00±3.39) days, respectively. Compared with the group A, the MST of group B was significantly longer (P < 0.05), and the MST of grafts in group D was also significantly longer than group C (P < 0.05). ③The expression of IFN-γ and IL-2 proteins was absent in group B. Compared with group C, the expression of IL-2 and IFN-γ proteins in group D significantly decreased on days 3, 7, 10, and 14 after keratoplasty (P < 0.05). CONCLUSION: Cervical lymphadenectomy therapy can effectively inhibit corneal allograft rejection in normal and high-risk corneal beds after alkali burn injury.

Objective To investigate the value of extravascular lung water (EVLW) in predicting the prognosis of sepsis acute lung injury (ALI) / acute respiratory distress syndrome (ARDS).Methods Relevant published articles concerned with EVLW both in Chinese and in English from 1991-2011 were searched,and Meta-analysis was carried out with Stata software. Results Seven articles were selected for Meta-analysis according to the inclusion and exclusion criteria.The results indicated EVLW of dead patients group kept in a high level,while EVLW showed a down-trend in the survival group.The difference in level of EVLW between the two groups became more obvious with the day ( SMDd1＜ SMDd2 ＜ SMDd3 ) ( the 1st day:SMDd1=0.29,95％CI:0.047-0.532; the2nd day:SMDd2=1.64,95％CI:0.14-3.13; the 3rd day:SMDd3 =1.83,95％ CI:0.56-3.10).Conclusions EVLW and its dynamic change in the early stage of sepsis ALI/ARDS could predict the prognosis.This suggested the sustained high level would lead to high mortality.

Objective To investigate the relationship between the serum high sensitive c-reactive protein (hs-CRP) and prealbumin (PAB) and the acute heart failure during the early stage of acute myocardial infarction (MI) patients.Methods A total of 181 MI patients (male:n =154,85.1％ vs.female:n =27,14.9％),admitted between Seppember 2010 and September 2012,were enrolled.They were divided into heart failure group (n =114,63.0％) with Killip classification recorded and control group (n =67,37.0％) without heart failure.The levels of serum hs-CRP and PAB were determined from the venous blood in the followed morning after admission.The clinical data were analyzed by logistic regression,Spearman correlation,and ROC curve.Results The serum level of hs-CRP (mg/L) in the heart failure group was significantly higher than that in the control group (P =0.000),while the serum level of PAB (mg/L) in the heart failure group was significantly lower than that in the control group (P =0.000).High level of hs-CRP and low level of PAB were significantly correlated with Killip classification (rhs-CRP =0.234,Phs-CRP =0.003 ; rPAB =-0.321,PPAB =0.000).Serum hs-CRP (P =0.023,OR 1.086,95％ Cl 1.012-1.167) and PAB (P =0.038,OR O.991,95％ CI O.983-0.999) were the independent risk biomarkers of acute heart failure subsequent to myocardial infarction determined by multivariate logistic regression analysis.The area under the ROC curve:AUChs-CRP =0.722,95％ CI 0.651-0.786; AUCPAB =0.723,95％ CI 0.652-0.787.Conclusions With high level of serum hs-CRP or low level of serum PAB during the early stage of acute myocardial infarction,patients were predisposed to the development of acute heart failure consequently.Both of them are the independent risk biomarkers of acute heart failure subsequent to myocardial infarction.Furthermore,they were significantly correlated with severity of the heart failure in terms of Killip classification.

Objective To investigate the value of plasma soluble triggering receptor expressed on myeloid cells-1 as a diagnosis marker of sepsis.Methods Articles on plasma soluble triggering receptor expressed on myeloid cells-1 as a marker of sepsis which were public published in the PubMed,Ovid,Springer,Wanfang database from 1991-2012 were searched and conducted a meta-analysis by MetaDiSc and Stata.Results Seven articles were selected to the meta-analysis according to the inclusion criteria,of which cut-off values varied signicantly from studies.Due to the data heterogeneity (I2 ＞ 50％,P ＜0.05),random model was used to pool the effect sizes.The overall combined effect sizes:sensitivity =81％ (95％CI:0.76-0.86); specificity =81％ (95％ CI:0.76-0.86); DOR =30.03 (95％ CI:7.89-114.37) ; AUC of SROC =0.905 9; Q*-0.837 6.Deek' s funnel plot showed little publication bias.Conclusions Plasma soluble triggering receptor expressed on myeloid cells-1 may be a useful adjunctive tool for the diagnosis of sepsis.However,further studies are needed in order to identify the best cut-off value in the diagnosis of sepsis.

Heat-shock protein (Hsp) 70 potentiates specific immune responses to some antigenic peptides fused to it. Here, the prokaryotic plasmids harboring the envelope glycoprotein E0 gene of classical swine fever virus (CSFV) and/or the Hsp70 gene of Haemophilus parasuis were constructed and expressed in Escherichia coli Rosseta 2(R2). The fusion proteins were then purified. Groups of Balb/c mice were immunized with these fusion proteins, respectively, and sera collected 7 days after the third immunization. Immune effects were determined via an enzyme-linked immunosorbent assay and flow cytometric analyses. E0-Hsp70 fusion protein and E0+Hsp70 mixture significantly improved the titer of E-specific antibody, levels of CD4+ and CD8+ T cells, and release of interferon-γ. These findings suggested that Hsp70 can significantly enhance the immune effects of the envelope glycoprotein E0 of CSFV, thereby laying the foundation of further application in pigs.
Animals ; Antibodies, Viral ; blood ; CD4-Positive T-Lymphocytes ; cytology ; immunology ; CD8-Positive T-Lymphocytes ; cytology ; immunology ; Cell Proliferation ; Classical swine fever virus ; genetics ; Female ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Haemophilus parasuis ; genetics ; Immunization ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics