Objective To investigate the effects of allergens on the expressions of IL-18,IL-18BPa and IL-18Rα protein in peripheral blood CD4+Th1 cells of healthy control subjects(HC)and patients with allergic rhi-nitis(AR),and on the expressions of IL-18,IL-18BPa and IL-18Rα mRNA in the peripheral blood CD4+T cells.Methods Blood samples were collected from patients with rhinitis for negative skin prick test(AR-),rhinitis for positive skin prick test(AR+)and HC.Flow cytometry was used to evaluate the effects of allergens on the expres-sions of IL-18,IL-18BPa and IL-18Rα protein in CD4+Th1 cells.The expressions of IL-18,IL-18BPa and IL-18Rα mRNA in CD4+T cells were determined by qPCR.Results Compared with HC,increased IL-18 while de-creased IL-18BPa expressions in Th1 cells of AR-and AR+patients were observed,increased IL-18Rα expression in Th1 cells of AR+patients was also found.Additionally,allergens induced elevated expression of IL-18Rα pro-tein in Th1 cells of HC,and induced elevated mRNA expressions of IL-18,IL-18BPa and IL-18Rα in isolated blood CD4+T cells of AR+patients and HC.Conclusion Allergens may be involved in the pathogenesis of AR by inducing the expressions of IL-18 and IL-18Rα in blood CD4+Th1 cells.

Objective Rapid elevation of the IgG antibody against Candida Enolase ( Eno ) has been observed in patients with invasive candidosis in an early stage .The present study was to confirm the association of Candida colonization with humoral im-mune anamnestic response of the dominant antigen . Methods Twenty-four mice were randomized into group 1 treated by oral Candi-da colonization plus intraperitoneal infection ( immunocompetent , n=8) , 2 treated with immunosuppressant in addition to the treatment of group 1 ( immunocompromised , n=8) , 3 treated by oral Candida colonization only ( immunocompetent , n=4) and 4 treated by in-traperitoneal injection only( immunocompetent, n=4).The number of Eno-specific memory B-cells in the spleen and the levels of IgG , IgM and IgA antibodies were determined in the peripheral blood of the immunocompetent and immunocompromised invasive candidiasis mice . Results At 7 days after invasive infection , there were significantly more Eno-specific memory B-cells in the mice of groups1 ( 47.25 ± 13.81) and 2 (43.14±15.95) than in groups 3 (8.00±3.74) and 4(8.50±2.38) (P<0.01), with no statistically significant differences between either groups 1 and 2 or groups 3 and 4 (P >0.05).Eno-IgG antibodies were detected in the serum of the mice of the first two groups in the early stage of invasive infection and positively corre -lated with antigen-specific memory B-cells (r=0.737,P <00.1 ). Conclusion Rapid elevation of the Eno-IgG antibody level in the early stage of invasive infection after Candida colonization may be attributed to the rapid proliferation of humoral immune memory cells.

Objective Invasive candidiasis is associated with a significant mortality clinically.The purpose of this study was to observe the T cells immune response to fructose bisphosphate aldolase (Fba), an immunodominant antigen of Candida albicans, and determine whether the antigen has the possibility of priming cellular immune protection in invasive candiasis.Methods Using ELISPOT assay, we determined the frequencies of positive spot-forming cells (SFCs) of Fba antigen-specific T cells secreting IFN-γ, IL-4 and IL-17A in the peripheral blood mononuclear cells (PBMCs) of 26 healthy individuals.Results After Fba stimulation, the frequencies of positive SFCs of IFN-γ, IL-4 and IL-17A in the 26 healthy subjects were 23 (9.75, 42.50), 0 (0, 0.25) and 1.5 (0.75, 8.25), respectively, with statistically significant differences among the three (P<0.01).The response rates of IFN-γ (100% [26/26]) and IL-17A (76.92% [20/26]) were significantly higher than that of IL-4 (15.38% [4/26]) (P<0.01).Fba-induced strong response (SCFs ≥20) for IFN-γ was observed in 57.69% (15/26) of the healthy individuals, that for IL-17A in only 1, while that for IL-4 in none.Responses of both Th1 and Th17 cells to Fba were found in 65.38% (17/26) of the subjects, that of Th1 cells in 19.23% (5/26), but that of Th2 cells in none.Conclusion Fba of Candida albicans can induce immunodominant responses of Th1 and Th17 cells and is a potential vaccine against invasive candiasis.

Objective To establish an ELISA for detecting antibody against Aspergillus fumigatus pectate lyase A(anti-PlyA),and evaluate its diagnostic value for invasive aspergillosis (IA).Methods An indirect ELISA for IgG antibody a-gainst PlyA was established using PlyA as coating antigen.The serum from 97 IA patients,80 non-IA patients and 200 healthy donors were tested,the results were compared with anti-DPPV (antibody against Aspergillus fumigatus dipeptidyl peptidase V fragment)and anti-TR (antibody against Aspergillus fumigatus thioredoxin reductase).Results The intra-as-say coefficients of variation of the ELISA method for detecting anti-PlyA was 5.3%,and inter-assay coefficients of variation was 10.9%.The sensitivity and specificity of diagnosis of IA were 62.9% and 90.4%,respectively.The positive rates of an-ti-PlyA in non-neutropenic and neutropenic IA patients were 43.8% and 72.3%,respectively (χ2 =7.493,P <0.05).There was no significant difference between the positive rates of anti-PlyA (62.9%),anti-TR (60.8%),and anti-DPPV (67.0%) (χ2 =0.562,P > 0.05).When combined anti-PlyA,anti-TR,and anti-DPPV,the diagnostic sensitivity for IA patients in-creased to 92.8%.Conclusion An ELISA for detecting anti-PlyA was successfully established.The diagnostic value of these three kinds of antibody was superior in non-neutropenic IA patientsto that in neutropenic IA patients.The combined detec-tion of three antibodies could provide higher sensitivity.

Objective The systemic inflammatory response syndrome ( SIRS) can be caused by infection and non-infection factors, which have similar clinical features but differ in treatment and prognosis .Rapid synthesis of procalcitonin ( PCT) during infec-tion can be used as a biomarker for the early diagnosis of sepsis .The present study aims to assess the value of the serum PCT level in the etiological diagnosis and prognosis of SIRS in the surgical ICU . Methods We retrospectively analyzed the data on 166 cases of SIRS from the surgical ICU in Jinling Hospital between June 2014 and June 2015 .The data obtained were associated with the patients'demograph-ics, primary diseases, laboratory results, and clinical outcomes.We analyzed the serum PCT values , blood culture results , and clinical outcomes. Results Totally, 131 of the patients were diagnosed with sepsis, with a median value of serum PCT of 2.43 (0.81-10.51) ng/mL, of whom 109 were PCT-positive (≥0.47 ng/mL), with a positive rate of 83.2%.Among the 35 non-infection SIRS patients, the mean level of serum PCT was 0.23 (0.1 -0.39) ng/mL, with a positive rate of 17.14%(6/35).There were statistically significant differences between the two groups in both the ser-um PCT level and positive rate (P<0.05).The PCT-positive rate was significantly higher in the bacteria-infected than in the fungi-in-fected group (86.5%[83/96] vs 74.3%[26/35], P<0.05), with a median value of 4.28 (1.05-14.59) ng/mL and 0.89 (0.37-1.59) ng/mL, respectively, so was it in the survivors than in the non-survivors (94.4%[34/36] vs 78.9%[75/95], P<0.05), with a median value of 12.89 (4.76-47.73) ng/mL and 1.41 (0.54-4.00) ng/mL, respectively. Conclusion The se-rum PCT level might be used to distinguish between sepsis and non-infection SIRS, significantly higher in bacteria-infected and survival groups than in fungi-infected and non-survival groups .Serum PCT determination contributes to the etiological diagnosis and prognosis of SIRS.

Objective To develop an immunomagnetic bead ( IMB) assay for quantitative detection enolase ( Eno) of Candi-da albicans, and to improve the diagonosis of invasive candidiasis . Methods The immunomagnetic bead was prepared by conjuga-ting with Anti Eno of Candida albicans monoclonal antibody .HRP-conjugated goat polyclonal antibody against Candida albicans Eno was employed as detecting antibody .The performance parameters of the IMB assay including precision , specificity, linear range and limit of detection were verified by using recombinant Candida albicans Eno.Then the developed assay was applied to determine Eno levels in supernatant of pathogenic fungi cultures . Results The intra and inter-coefficient of variation was 4.54%, 5.87% and 5.26%, 8.82%at the concentration of 25 ng/mL and 5 ng/mL, respectively.The limit of detection was 0.5 ng/mL.The linear range was(0.5-50) ng/mL.The level of Eno in Candida albicans culture after incubated in 37℃for 24 h was 3.89 ng/mL and gradually in-creased to 37.89 ng/mL at 120 h.There was a positive correlation between the level of Eno and growth hyphae of Candiad albicans. There was weak cross reaction with Candida parapsilosis and no cross reaction with Candida tropicalis, Candida guilliermondii, Candida glabrata, Cryptococcus neoformans and Saccharomyces cerevisiae. Conclusion An IMB assay for quantitative detection Eno of Candida albicans was developed , which was more sensitive , rapid and reliable than previous qualitative ELISA .The IMB assay has the potential to be applied to the research in invasive candidasis .

Objective The purpose of this study was to evaluate the protective potential of the Aspergillus fumigatus thiore -doxin reductase GliT ( TR) antigen by establishing and optimizing ELISPOT assay for TR antigen-specific T cells ( TR/AST) secreting IFN-γand IL-4 in peripheral blood mononuclear cells ( PBMCs ) and explore the role of TR/AST in invasive aspergillosis ( IA ) . Methods We optimized the reaction conditions of ELISPOT by preliminary checkerboard titration and determined the frequencies of positive spot-forming cells ( SFCs) specifically secreting IFN-γand IL-4 in the PBMCs of 20 healthy individuals with TR as specific stimulant and with PHA and PMA as positive controls ,. Results Checkerboard titration demonstrated the best result of ELISPOT with the TR antigen at the final concentration of 10μg/well and PBMCs at 3 ×105/well.The median frequency of IFN-γSFCs was sig-nificantly higher (15 [3.5, 59.5]) than that of IL-4 SFCs (0 [0, 0]) (P<0.001).TR induced IFN-γresponses in all the 20 healthy donors, including 9 cases of strong IFN-γresponse (SFCs>20/3 ×105 PBMCs), accounting for 45%, but failed to induce IL-4 response in 19 of the healthy individuals . Conclusion The Aspergillus fumigatus TR antigen could induce an immunodominant Th1 response , and therefore might be a potential protective antigen .

An immunochromatographic Lateral-Flow Device (LFD)was invented for rapid serodiagnosis of Invasive Asper-gillosis by foreign investigator.A monoclonal antibody against Aspergillus extracellular glycoprotein is used as capture anti-body and detection antibody in LFD.LFD is a simple,rapid,single sample performance technique by which Aspergillus ex-tracellular glycoprotein in serum and/or bronchoalveolar lavage fluid (BAL)of patients can be detected.The principle of LFD and the features of antigen and antibody involved in the technique will be described.The diagnositic value of novel LED is compared with that of GM test and PCR.

AIM: To prepare anti-Sperm protein 17 (Sp17) immunomagnetic nanoparticles (IMNPs), and make foundation for target diagnosis of ovarian cancer by magnetic resonance imaging. METHODS: The anti-human Sp17 IMNPs were prepared by grafting anti-Sp17 antibodies on the surface of chitosan-coated magnetic nanoparticles （MNPs） using the linker of EDC/NHS (1-ethyl-3-［3-dimethylaminopropyl］carbodiimide/N-hydroxysuccinimide). The morphology and properties of the nanoparticles were characterized by transmission electronic microscopy (TEM), the conjugation of the antibodies was evaluated by native-polyacrylamide gel electrophoresis, the immunologic activity of IMNPs was evaluated by enzyme linked immunosorbent assay (ELISA). A set of in vitro magnetic resonance imaging (MRI) experiments were performed after incubated the IMNPs with human Sp17 gene transfected ovarian cancer HO-8910 cells. RESULTS: We had successfully grafted the MNPs with anti-Sp17 antibody and the IMNPs kept good bioactivity. The MRI showed that the IMNPs were targeted successfully to the positive cells, and no obviously non-specific adsorption was observed. CONCLUSION: The anti-Sp17 IMNPs with good specificity can used for further study of ovarian cancer target therapy.

Objective To investigate whether rosuvastatin has the effects of anti-oxidative stress and attenuates vascular endothelial adhesiveness on the vessel wall in apolipoprotein E(apoE)knockedout mice. Methods Eighty 8-week-old apolipoprotein E-deficient mice and twenty 8-week-old C57BL/6 mice,were fed a normal chow diet for 12 weeks,and were divided into 2 weeks treatment group and 6 weeks treatment group.apoE knocked-out mice were subcutaneously injected with rosuvastatin at a dose of 1, 5, or 20 mg/kg daily for 2 or 6 weeks prior to sacrifice.Blood sample was taken by cardiopuncture and the aorta was obtained at the end of the 2th or 6th weeks. Results Total cholesterol level was significantly decreased after 2 or 6 weeks of 5,or 20 mg/kg rosuvastatin treatment[2 weeks:(480.7±35.3)mmol/L,(371.5±27.1)mmol/L;6 weeks:(400.1±37.6)mmol/L,(305.0±19.3)mmol/L],compared with 0 mg/kg group(20 weeks:(675.0±42.0)mmol/L;6weeks:(660.0±44.3)mmol/L](P＜0.05 or 0.01).But the changes of triglyceride and high-density lipoprotein were not like the above.The endothelial adhesiveness for monocytes was significantly attenuated after 2 weeks in 20 mg/kg group versus 0 mg/kg group[(2.24±0.72)%vs.(3.76 ±2.53)%](P＜0.05),and even more significantly after 6 weeks in 5,20 mg/kg groups[(1.94±0.40)%,(3.95±2.61)%](P＜0.01).In addition,rosuvastatin inhibited vascular expression of p22phox and superoxide production[0 mg/kg for 6 weeks group:(3.22±1.53)%,(4.75±2.62)μg/L;20 mg/kg for 6 weeks group:(1.41±0.72)%,(2.72±0.88)μg/L](all P＜0.05). Conclusions Rosuvastatin has a role of anti-oxidative stress and attenuates vascular endothelial adhesiveness on the vessel wall.