Objective To explore the CD4 + CD25 + Foxp3 + regulatory T cell percentage and plasma levels of soluble CD25 molecules in peripheral blood of septic patients and their clinical value through prospective study. Method A total of 37 septic patients and 15 non-infectious SIRS patients, who conformed to the criteria of SIRS and sepsis which proposed by American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference ( ACCP/SCCM ) in 1997, were collected in ICU of Ruijin Hospital ( Shanghai Jiaotong University) from February 2009 to February 2010. Twenty-four health people were from Medical Center of Ruijin Hospital, who were excluded infection and (or) autoimmune diseases. There were 26 male and 11 female in sepsis group, average age ( 61.67 ± 11.87 ) years old; 8 male and 7 female in SIRS group, average age (67.06 ± 12.57)years old; 14 male and 10 female in health control, average age (56.54 ± 6.37 )years old. All selected patrents were excluded the autoimmune diseases and (or) patients within recent (30 days) had used or now used immunosuppressive agents. We therefore measured the Treg cell percentage in peripheral blood by Flow Cytometry and the plasma levels of IL-2sRa, IL-4, IFN-γ by ELISA. The data were analyzed by analysis of variance or nonparametric Kruskal-Wallis H test. Results ① The percentage of CD4 + CD25 + Foxp3 + regulatory T cells among septic patients, SIRS patients, and control group was: ( 66.82 ± 21.79 ) %, ( 51.79 ± 21.79 ) %, ( 56.45 ± 10. 68 ) %, respectively. septic patients showed the highest percentages of CD4 + CD25 + Foxp3 + regulatory T cell among CD4 + CD25 + T cells(P ＜ 0.05 ). ② The plasma levels of soluble CD25 in septic patients (425. 619 ± 270.12 ) were significantly higher than SIRS patients (381. 664 ± 189.83) and the control group ( 164. 1 32 ± 56.37 ) ( P ＜ 0.05 ). ③ The correlation analysis between the concentration of soluble CD25 molecules in plasma and the ratio of CD4 + CD25 + Foxp3 + regulatory T cells to CD4 + CD25 + T cells showed Spearman correlation coefficient =0.390, P = 0.003 ( P ＜ 0.05 ). Conclusion: the expression of natural regulatory T cells characteristically increased in septic patients. And the levels of soluble CD25 in peripheral blood were related to the percentages of natural regulatory T cells, which simplified the assessment of the immune status in Septic patients.

Objective To explore the mRNA expressions of NLRP1 (NOD-like receptor 1 ) and NLRP3 inflammasomescomponents and related pro-inflammation cytokines in peripheral blood mononuclear cells (PBMCs).Methods A total of 21 septic patients,20 non-infectious SIRS patients and 20 health subjects were enrolled in this study.The mRNA expressions of inflammasomes components and related proinflammation cytokines were measured by using RT-PCR,and serum levels of IL-1β and IL-18 were measured using ELISA.One-way ANOVA was usedd to compare the data of all three groups,and SNK-q was employed to compare thedata among different groups.Spearman correlation analysis was used to analyze the correlation between these biomarkers and the severity of septic patients.Results ①The mRNA expression of NLRP1 in PBMCs of septic patients in early stage and non-infectious SIRS patients was down-regulated in comparison with healthy subjects (P ＜0.01 ).The expression of NLRP3 in septic patients was not differentfrom that in healthy subjects ( P ＞ 0.05 ),but NLRP3 mRNA expression in non-infectious SIRS patients was significantly higher than that in septic patients and in healthy subjects ( P ＜ 0.01 ).ASC ( apoptosis associated speck like protein containing a CARD) and caspase-1 mRNA expressions in septic patients and non-infectious SIRS patients were higher than those in healthy subjects ( P ＜ 0.01 ).There was no difference in level of IL-1β found among all groups.IL-18 mRNA expression in septic patients and non-infectious SIRS patients was higher than that in healthy subjects ( P ＜ 0.01 ),and that in non-infectious SIRS patients was highest.②)Serum protein level of IL-1β in septic patients was lower than that in healthy subjects (P ＜0.05 ),and serum level of IL-18 in septic patients was significantly higher than that in non-infectious SIRS patients and in healthy subjects ( P ＜ 0.01 ).③NLRP1 was negatively related to SOFA ( sepsis-related organ failure assessment) score (r =-0.44,P ＜ 0.05 ) and negatively related to APACHE Ⅱ score as well ( r =- 0.52,P ＜ 0.05 ).Conclusions NLRP1 mRNA expression was down regulated in patients with early sepsis and its expression was negatively correlated to the severity of septic patients,indicating the lower NLRP1 mRNA expression the severer illness.

Objective To evaluate antibiotic therapy for the treatment of intraabdominal infection secondary to severe acute pancreatitis.Methods 39 SAP patients with intraabdominal infection were devided into 2 groups: early infection group (early group) and late infection group (late group) according to the onset time of infection. All patients received imipenem and or surgical treatment, and results were compared between the two groups. Results 83 strains of pathogen were isolated among which Gram negative bacteria accounting for 84%, with a ratio of sensitivity to imipenem of 80%; Gram positive bacteria taking up 16%, all were resistant to imipenem. The cure rate was 53% in eraly group and 86% in late group(P=0.02); The rate of bacteria clearance was 47% in early group and 82% in late group(P=0.02). The survival rate of patients in early group was 77% and in late group was 95%(P=0.08); The reinfection rate in early group was 58% and in late group was 37%(P=0.16). Conclusion Imipenem is effective in the treatment of intraabdominal infection secondary to severe acute pancreatitis,and repeated surgical intervention is still necessary for patietns in early infection group.

Objective To investigate the prevalence and main genotypes of carbapenemases in carbepenem‐resistant Enterobacteriaceae (CRE) .Methods A total of 114 strains of CRE were isolated in Shanghai Ruijin Hospital from May 2011 to June 2013 .The diameter of inhibition zone of imipemen or meropenem for these strains was not larger than 22 mm .PCR method was used to screen for the main carbapenemase genes (blaKPC ,blaIMP ,blaVIM ,blaOXA‐48 and blaNDM ) with previously described primers followed by nucleotide sequencing analysis . Conjugation experiments were performed to examine the transferability of plasmids .Pulsed‐field gel electrophoresis (PFGE) was used to show the relatedness of KPC‐2‐producing Enterobacteriaceae .Results Most of the 114 isolates were K lebsiella pneumoniae and Escherichia coli .Of the 114 isolates ,98 was positive for carbapenemases ,specifically ,78 blaKPC‐2‐positive ,15 blaIMP‐4‐positive ,2 blaIMP‐8‐positive ,1 positive for both blaKPC‐2 and blaIMP‐4 and 4 blaNDM‐1‐positive .None of the strains was positive for blaOXA‐48 or blaVIM .About 21 .4% (21/98) of the isolates were conjugated successfully .The 49 blaKPC‐2‐positive K .pneumoniae isolates were grouped into 12 types according to PFGE patterns .Majority (34/49) of these isolates belonged to the same type A .Conclusions BlaKPC‐2 was the primary epidemic genotype of Enterobacteriaceae in Ruijin Hospital ,followed by blaIMP‐4 .NDM‐1 carbapenemase was produced in 4 strains of CRE . Meanwhile , clonal spread of KPC‐2‐producing K . pneumoniae was observed in some departments of our hospital , such as surgical ICU , respiratory medicine and thoracic surgery . Appropriate measures should be taken timely and effectively to prevent the in‐hospital spread of resistant genes .

Objective To investigate the diagnostic value of serum proadrenomedullin( pro-ADM)in catheter related blood stream infection( CRBSI). Methods Prospective diagnostic test was performed with 76 cases patients with clinically suspected diagnosis of CRBSI,and the patients were divided into CRBSI group( n=25)and non-CRBSI group. Serum pro-ADM,procalcitonin( PCT),C-reactive protein( CRP)and white blood cell( WBC)levels were measured at the study entry and on the day of CRBSI suspicion. Results On the day of CRBSI suspicion,the levels of pro-ADM,PCT,CRP and WBC in the CRBSI group were(5. 17 ± 1. 28)nmol/L, 2. 29(1. 47-4. 28)μg/L,(102. 04 ± 51. 00)μg/L and(14. 66 ± 5. 09)× 109/L respectively,significantly higher than those in the non-CRBSI group((2. 83 ± 1. 25)nmol/L,2. 29(1. 4-4. 28)μg/L,(61. 43 ± 53. 52) mg/L and(11. 78 ± 3. 52)× 109/L respectively;t or Z=7. 636,-4. 777,3. 156,2. 882;P﹤0. 05). For the diagnosis of CRBSI,the area under the curve(AUC)of pro-ADM,PCT,CRP and WBC were 0. 89(95%CI 0. 82-0. 97),0. 84(95%CI 0. 75-0. 93),76(95%CI 0. 65-0. 86)and 0. 68(0. 54-0. 81)respectively. Taking pro-ADM with 4. 31 nmol/L as the cutoff value,the sensitivity,specificity,positive predictive value and negative predictive value were 76. 0%,84. 3%,70. 4% and 87. 8% respectively. Simultaneously,taking procalcitonin with 1. 52 μg/L as the cutoff value, the sensitivity and specificity was 81. 8% and 87. 0%respectively. Conclusion Serum level of pro-ADM in the diagnosis of CRBSI has good specificity. Simultaneous surveillance of serum pro-ADM and PCT may be helpful for the diagnosis of CRBSI in the early stage.

Objective To analyze the change in the concentration of IL-17 and IL-10 inflammatory factors among the deadaptation personnel who returned from the plateau.Methods A total 21 healthy males were investigated who averaged 25 years in age, lived permanently in the plains (200 m), and once stayed to the plateau (Lasha) for 6 months.Their venous blood was collected at three time points:the day before ascending to the plateau(control), the second day after return to the plains(d2) and the 30th day(d30), respectively.Their serum was seperated from the whole blood and the level of IL-17A and IL-10 was detected by ELISA method.Results The concentration of IL-17A and the IL-17A/IL-10 ratio were significant increased at d2 and d30, respectively, compared with control (P<0.05).Compared with d2, IL-17A and the IL-17A/IL-10 ratio were decreased obviously at d30(P <0.05).The level of IL-10 at d2 and d30 was significantly reduced compared with control ( P <0.05), but increased at d30 compared with d2.Correlative analysis showed that there was a negative correlation in the levels of IL-10 and IL-17A between control, d2 and d30, respectively (r1=0.948, P<0.05;r2=0.969, P<0.05;r3=0.972, P<0.05).A significant negative correlation was observed in the alteration levels of IL-10 and IL-17A between the three groups(r4=-0.793, P<0.05; r5=-0.756, P<0.05). Conclusion The concentration of inflammatory factors among the plateau deadaptation patients is imbalanced, but it is gradually reduced with time.The mechanism is still not clear.

Objective To analyze the MRI features of central neurocytoma (CNC),and to improve the diagnostic accuracy of this lesion.Methods The MRI findings of 12 patients with CNC confirmed by surgery and pathology were evaluated retrospectively.All patients underwent contrast-enhanced and uncontrast-enhanced MR scan,and 3 patients also received plain CT scan.Results The lesions of 12 cases were all located in the lateral ventricles of septum pellucidum,which were close to the Monro's foramen.5 lesions located in the right lateral ventricle,5 lesions located in both lateral ventricles with 1 case reached the third ventricle,2 lesions located in the left lateral ventricle with 1 case reached the third ventricle.The mean diameter of the CNC lesions was 4.8 cm (ranged from 2.2 to 7.8 cm).11cases (92％) had vavious degrees of cystic change,8 cases (66.7％) had a spongy or soap bubble appearance and 1 case(8.3％) had completely cystic change.3 cases (25％) had a scalloping appearance on sagittal T1-weighted MRI.The solid part of the tumor was similar to the gray matter in signal intensity,with slightly high signal intensity on T2FLAIR.Among 9 cases with DWI,7 cases showed high signal intensity,1 case showed equal signal intensity,and 1 case showed slightly low signal intensity.On enhanced MRI,the solid part of tumors showed slight to medium (83％) or obvious (17％) enhancement.Conclusion CNC should be first considered when a young adult has a lesion located in the lateral ventricles of septum pellucidum,accompainied by varying degrees of cysts,showed slight to medium enhancement in the solid part of the tumor.

Objective To study the MRI features of primary central nervous system lymphoma (PCNSL)to improve its diagnosis. Methods The MRI data of 1 3 patients with PCNSL confirmed by biopsy or surgical pathology were analyzed retrospectively.All patients underwent nonenhanced and contrast-enhanced MRI examinations.Results Multiple lesions in 8 cases and solitary one in 5 cases were found.Five leisons (38.5%)were located in the cerebral hemispheres.Eight(61.5%)were located in deep structures of the brain including periventricular white matter,basal ganglia,thalami and corpus callosum,6 (46.2%)of whom involved in the ependyma including 1 involved in the ependyma of the 4 th ventricle.One leison(7.7%)was located in the cerebellar hemisphere.The tumor showed isointense or mildly hypointense signal on T1WI,mildly hyperintense signal on T2WI and hyperintense signal on DWI.Mild to moderate perilesional edema was noted.The tumors in 13 cases showed remarkable homogeneous enhancement after the contrast agent was injected with the signs of characteristic notch,butterfly and satellite.Focal necrosis occurred in 1 lesions.Conclusion PCNSL shouldbe first considered when a lesion located in the deep central structeres of the brain,multiple and involve ependymal membrane,showed isointensity on T1WI and T2WI,highintensity on DWI in tumoral parenchyma demonstrated,significantly enhancement in the solid part of the tumor.

The current situation of multidrug-resistant bacteria infection in surgical patients is becoming increasingly severe.Effective prevention of cross transmission of multidrug-resistant bacteria and common site infections in ICU patients is the key step.In response to the key issues in the prevention and control of multidrug-resistant bacteria infection,a comprehensive strategy is implemented.Core measures such as early recognition,preemptive isolation,active screening,and graded prevention and control are taken for patients,and targeted preventive measures for common infection sites are combined to form a detection and control system of multidrug-resistant bacteria for ICU patients.By moving forward and extrapolating to the general ward,the prevention and control system effectively reduced the incidence of multidrug-resistant bacteria colonization/infection in surgical patients.

OBJECTIVETo develop and identify the monoclonal antibodies (mAb) against c-erbB-2 p185 intracellular domain for detection of c-erbB-2 protein overexpression in breast tumor cells.

METHODSBALB/C mice were immunized with a synthesized p185 peptide of intracellular domain. The biological characteristics and immunoactivities were identified by different techniques.

RESULTSThree hybridoma cell strains secreting mAbs to c-erbB-2 protein were established and one of the mAbs, No. 035-E61 was tested for c-erbB-2 protein immunostaining on 39 breast carcinoma sections, 30 breast fibroadenoma sections and 16 sections from various normal organs. The results showed that the positive detection frequency of protein expression was 26% (10/39) in breast cancer, none (0/30) in fibroadenoma and 3 sections from normal organs were positively stained. The consistency between the 035-E61 and the DAKO reagent approved by FDA was 74%.

CONCLUSIONSNo. 035-E61 mAb can specifically recognize the p185 intracellular domain and may be useful in guiding clinical Herceptin treatment.

Animals ; Antibodies, Monoclonal ; immunology ; Antibody Specificity ; Binding Sites ; immunology ; Breast Neoplasms ; immunology ; pathology ; Female ; Fibroadenoma ; immunology ; pathology ; Humans ; Immunoglobulin G ; immunology ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Receptor, ErbB-2 ; immunology ; Tumor Cells, Cultured