Objective To screen the specific cytokines of tuberculous pleural effusion(plTB)by using liquid array technique to establish a diagnostic model and discuss its application value.Methods A total of 86 patients with plTB(plTB group)were included,including 41 patients in the confirmed plTB group and 45 patients in the clinically diagnosed plTB group.There were 42 other patients with pleural effusion in the control group.Seventeen cytokines in pleural effusion were analyzed by liquid array technology.Interleukin(IL)-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-9,IL-10,gamma-interferon-induced protein 10(IP-10),IL-15,IL-17F,IL-27,tumor necrosis factor(TNF)-α,monocyte chemotactic protein-1(MCP-1),the expression levels of macrophage inflammatory protein-3a(MIP-3α),macrophage colony-stimulating factor(M-CSF)and β-interferon(IFN-β)were detected.Difference factors between the confirmed plTB group and the control group were screened,and the receiver operating characteristic(ROC)curve was drawn in the confirmed plTB patients.IP-10,IL-27 and MCP-1 with AUC>0.850 and specificity>80%were combined to diagnose plTB,and were compared with adenylate deaminase(ADA)and T-SPOT.TB in pleural effusion to evaluate the diagnostic efficacy.Results The levels of IL-2,IP-10,IL-27,TNF-α and MCP-1 were higher in the confirmed plTB group than those in the control group(P＜0.05).The sensitivity and specificity of IP-10,IL-27 and MCP-1 in the diagnosis of plTB were 87.8%and 81.0%.The sensitivity of three-factor combined diagnosis in 45 patients with plTB was still as high as 86.7%,and there was no significant difference in sensitivity compared with that in the diagnosed plTB group(P＞0.05).In the plTB group,the sensitivity of IP-10,IL-27 and MCP-1 combined detection was 87.2%,which was higher than that of T-SPOT.TB(81.4%)and ADA(54.7%).Conclusion The application of liquid array technology to the joint detection of pleural effusion IP-10,IL-27 and MCP-1 can provide help for the diagnosis of plTB.

Objective To measure the expression of BP180 antibody in sera of patients with bullous pemphigoid (BP) and/or nervous system diseases (ND),and to explore the relationship between BP and ND.Methods Clinical data were collected from some inpatients and outpatients with BP in Department of Dermatology,as well as some inpatients with ND in Department of Neurology,the Second Hospital of Jilin University between March 2012 and September 2013.Peripheral blood samples were obtained from 20 BP patients without a medical history of ND (BP group),20 patients with ND alone (ND group),20 BP patients with a medical history of ND (BP + ND group),and 20 healthy controls (healthy control group) separately.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum level of BP180 antibody in the above groups.Statistical analysis was carried out with SPSS17.0 software by using chi-square test for comparing enumeration data,analysis of variance for comparing the anti-BP180 antibody titer or ages among the above 4 groups,and Games-Howell test for multiple comparisons.Results No anti-BP180 antibody was detected in any of the 20 subjects in the healthy control group.All of the patients in the BP group and BP + ND group were positive for anti-BP180 antibody,and the antibody titers were 128.347 ± 54.678 and 143.482±72.568 respectively.Of the 120 patients in the ND group,7 were positive for anti-BP180 antibody,including 4 with cerebral hemorrhage and 3 with cerebral thrombosis,and the highest anti-BP180 antibody titer of 39.638 was recorded in 1 patient with cerebral infarction.There was a significant difference in the anti-BP180 antibody titer among the 4 groups (F =55.624,P ＜ 0.05).In addition,there was no significant difference in the anti-BP180 antibody titer between the BP group and BP + ND group (P =0.878),while the anti-BP180 antibody titers significantly differed between the BP group and ND group,between the BP group and healthy control group,between the BP + ND group and ND group,between the BP + ND group and healthy control group,and between the ND group and healthy control group (P ＜ 0.05).Conclusion BP may occur in ND patients with an increased anti-BP180 antibody titer.

Objective To analyze the epidemiological characteristics and spatial distribution of human brucellosis in Fujian province during 2011-2016,and provide evidence for the prevention and control of the disease.Methods The surveillance data of human brucellosis in Fujian during 2011-2016 was analyzed with software R 3.3.1,ArcGIS 10.3.1,GeoDa 1.8.8 and SaTScan 9.4.3.Results During 2011-2016,a total of 319 human brucellosis cases were reported,the incidence increased year by year (F=11.838,P=0.026) with the annual incidence of 0.14/100 000.The male to female rate ratio of the incidence was 2.50 ∶ 1.Farmers and herdsmen accounted for 57.37％.The incidence was 0.40/100 000 in Zhangzhou and 0.32/100 000 in Nanping,which were higher than other areas.The number of affected counties (district) increased from 12 in 2011 to 28 in 2016,showing a significant increase (F=13.447,P=0.021).The Moran' s I of brucellosis in Fujian between January 2011 and December 2016 was 0.045,indicating the presence of a high value or low value clustering areas.Local spatial autocorrelation analysis showed that,high-high clustering area (hot spots) were distributed in Zhangpu,Longhai,Longwen,etc,while high-low clustering areas were distributed in Nan' an and Jiaocheng,etc.Temporal scanning showed that there were three clustering areas in areas with high incidence,the most possible clustering,occurring during January 1,2013-December 31,2015,covered 6 counties,including Yunxiao,Pinghe,Longhai,etc,and Zhangpu was the center,(RR =7.96,LLR=92.62,P＜0.001).Conclusions The epidemic of human brucellosis in Fujian is becoming serious,and has spread to general population and non-epidemic areas.It is necessary to strengthen the prevention and control of human brucellosis in areas at high risk.

ObjectiveTo evaluate the IL-17 expression in HIV/tuberculosis-coinfected patients and its role in the pathogenesis of this coinfection.MethodsFifty-four HIV infected patients were divided into three groups:simple HIV infected group,HIV with latent tuberculosis infection (HIV+ LTBI) group and HIV coinfected with active tuberculosis (HIV+ ATB) group.The whole blood intracellular cytokine staining was performed and samples were then detected by BD FACSCanto.The expressions of CD4+ IL-17+ T cells and CD4+ IFNγ+ T cells were analyzed using FACSDiva software.Comparison between groups was done by independent sample t test.ResultsThe CD4+ T cell count and viral load among these three groups were comparable.There were no significant difference of the expression of CD4+ IL-17+ T cells between simple HIV infected group and HIV+ LTBI group (1.40 ± 1.01) ％ vs (1.29±0.86) ％,(t=0.336,P＞0.05),but both of these two groups were much higher than HIV+ATB group (t=3.680,t=2.516,P＜0.05).There were no significant differences of the expression of CD4+ IFNγ+ T cells among these three groups [(32.8±24.0)％ vs (40.3±1 21.9) ％ vs (46.1±31.2)％,(t=-0.939,t=-1.602,t=-0.646,P＞0.05)].ConclusionThe Th17 response is down-regulated in HIV/tuberculosis-coinfected patients,which may play an important antitubercular role in the pathogenesis of coinfection.

Objective To evaluate the diagnostic value of two tuberculosis-specific IFN-γ release assays in latent tuberculosis infection among HIV-infected individuals. Methods The levels of tuberculosis antigen-specific IFN-γin 102 HIV patients from AIDS Outpatient Clinic of Shenzhen Third People's Hospital were detected by in-house tuberculosis-specific IFN-γ ELISpot assay and commercial T-SPOT TB kit, and tuberculin skin test (TST) were done at the same time. There were 66 males and 36 females,and the average age was 35. Results Seventeen HIV infected patients were positive in both IFN-γ ELISpot and T-SPOT TB methods, the sensitivity, specificity positive predictive value(PPV), negative predictive value(NPV) and compliance rates of ELISpot were 94. 4% ,94. 0% ,77. 3% ,98. 8% and 94. 1% ,respectively. Three patients were positive in both IFN-γELISpot and T-SPOT TB methods, the sensitivity, specificity, PPV, NPV and compliance rates of TST were 16. 7%, 98. 8%, 75.0%, 84. 7% and 84. 3%, respectively. The average number of spots using three kinds of antigen ESAT-6, Pool A,Pool B obtained were 26. 89 ±5. 77,18. 96 ±4. 75 and 14. 51 ± 3.77, respectively. Only ESAT-6 and Pool B have a statistically significant difference (H=7.557,P = 0.022 9), no significant difference was shown between other groups. There was no significant difference between the positive rate and the CD4+ T cellls number(x2 =0. 860 8 ,P =0. 650 2) ,as the same as the T-SPOT TB (x2 = 1. 396 4, P = 0. 497 5 ). Conclusions The performance of this in-house tuberculosis-specific IFN-γ ELISPot assay was comparable to T-SPOT assay in diagnosis of latent tuberculosis infection, and the sensitivity and specificity of both these two assays were all much higher than TST. They canbe recommended in diagnosing latent tuberculosis infection in HIV infected patients.

Objective To investigate the distribution of high pathogenicity island(HPI)in multiple-drug-resistance gram-negative bacilli and analyze the protein sequence.Methods To amplify thefyuA-irp2 gene cluster of the 84 isolates by multiple polymerase chain reaction(PCR),the product was subsequently sequenced.Results The positive rate ofirpl,irp2,irp3,irp4 and fyuA was 40.48％,41,67％,5.95％,O％and 16.67％,respectively.Theamino sequence offyuA comefromEC06748,Kp7151 and PAE7 was usedto compare with AL590842,there are 100％identities.Amino sequence ofirp2 come from Kp49 and Kp51 have 99％identities with AAA27636.1,but amino sequence of irp2 come from EC04 and EC07 only have 90％identities with 1176840.The GenBank accession number is FJ211852 and FJ211851.Amino sequence ofirpl come fromKp 10,Kp49 and Kp51 have 99％identities with AL590842。and amino sequence ofirp3 come from EC03,Kp51,Kp10 and Kp49 have 97％identities with CAA73128.There are the same mutation among the same species,and different mutation among different species.Conclusion There was different extant mutant lost in thefy～t-i,v2 gene cluster in multiple-drug-resistanee gram-negative bacilli.

Objective To assess the validity of a newly developed in-house ELISPOT IFN-γ release assay (IGRA) for the detection of latent tuberculosis infection among HIV infected individuals. Methods In-house ELISPOT assay were performed, together with a tuberculin skin test in 205 health controls and 110 HIV infected individuals , who had no signs of active tuberculosis at time of enrolment . Results Using the ELISPOT assay, positivity rates for the 205 health controls, 110 HIV infected individuals and 47 AIDS patients on highly active antiretrovial therapy (HAART) were 7. 3% , 24.5% , 29. 8% , respectively. These results indicated that the positive rates obtained from HIV infected individuals (include patient on HAART) was significantly higher than health controls( P ＜ 0.001). We found no significant correlation between the CD4 cell count and positivity of ELISPOT assay (P ＞0.05 ). The proportion of subjects with a positive response to ELISPOT assay were higher than the proportion of tuberculin skin test(TST) responders(P＜0.0001) in HIV infected individuals. Conclusion Our study indicates that IGRA using M. tuberculosis specific antigens are likely to retain their validity for the diagnosis of LTBI among HIV positive individuals.

Objective To develop an ELISA(Enzyme-Linked Immunosorbent Assay)diagnostic kit for early rapid detection of sarum anti-EV71 antibody and evaluate its clinical application value.Methods Recombinant protein VP1 of EV71 were prepared and purified as an immobilized antigen for establishment of an indirect ELISA for detection of serum anti-EV71 IgM and anti-EV71 IgG.Compared with RT-PCR.isolation of EV71 and micro-neutralizing assay.the clinical application value of anti-EV71 IgM and anti-EV71 ISG in the diagnosis of EV71 disease was evaluated.Results In comparison with RT-PCR.the sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgM antibody were 83%,85%,81%and 87%,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgG antibody were 72%,74%,68%and 77%.respectively.Compared with viral isolation assay.the sensitivity and specificity of anti-EV71 IgM antibody were 85%and 97%,respectively.The sensitivity and specificity of anti-EV71 IsG antibody were 75%and 77%,respectively.In addition.the titers of anti-EV71 IgG antibody were significantly correlated with the titers of neutralizing antibody to EV71 by linear regression analysis(r=0.72,P＜0.05).Finally,the serum titers of anti-IgG from patients with EV71 associated hand food and mouth disease at convalescent stage exhibited significantly higher than that of the same patients at acute stage(P＜0.01),but the titers of anti-IgM had no significant difference(P＞0.05).Conclusions With VP1 recombinant protein used as an immobilized antigen,an indirect ELISA diagnostic kit was successfully develooed for detection of serum anti-human EV71 IgM and anti-human EV71 IgG antibodies.

OBJECTIVETo explore the association of the polymorphism of homologue of dendritic cell-specific intercellular adhesion molecule-3 (ICAM-3) grabbing nonintegrin (DC-SIGN related, DC-SIGNR) gene with the susceptibility to HIV-1 infection.

METHODSThe distribution of the DC-SIGNR variants in the tandem repeat region and their association with HIV-1 infection in a cohort composed of 345 HIV-1 seropositive and 468 high-risk HIV-1 seronegative individuals was examined.

RESULTSThere are 14 genotypes and 5 alleles in the DC-SIGNR repeat regions in the cohort. Although the most common DC-SIGNR allele among Chinese Han population and the Caucasian population is 7, it was found in a higher frequency in the Chinese than in Caucasians (67.1% vs.46.0%, P<0.01). HIV-1 seropositive individuals had a lower frequency of the genotype 7/7 than the high-risk seronegative individuals (38.55% vs. 48.29%, P=0.0057), but a higher frequency of genotype 9/5 (4.35% vs. 1.07%, P=0.0029).

CONCLUSIONThese results suggest that the tandem-repeat polymorphisms of the DC-SIGNR gene in the Chinese Han population exhibit unique genetic characteristics previously unrecognized in the Caucasian population. Genotype 9/5 seems to be a risk factor for HIV-1 infection in the Chinese population.

Adult ; Alleles ; Cell Adhesion Molecules ; genetics ; Cohort Studies ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; HIV Infections ; genetics ; transmission ; virology ; HIV-1 ; Humans ; Lectins, C-Type ; genetics ; Male ; Polymorphism, Genetic ; Receptors, Cell Surface ; genetics

Objective To investigate the clinical characteristics and imaging manifestations of AIDS complicated with disseminated Penicillium marneffei (PM) infection. Methods A total of 12 patients with AIDS complicated with disseminated PM infection were collected and the symptoms, signs, laboratory examination results and image manifestations of these patients were analyzed retrospectively. Results (1) The diagnosis of PM infection in all the 12 cases were confirmed by peripheral blood culture.All the 12 cases (100％) had irregular fever (38-41 ℃) and enlarged lymph nodes, 8 cases (66％) had skin rashes; 8 cases (66％) had hepatomegaly; 9 cases (75％) had splenomegaly while 8 cases (66％) had anemia. (2) Imaging manifestation: Five cases manifested bilateral pulmonary disseminated miliary nodular shadows or lattice signs; 1 case showed enlarged hilar lymph node and 2 zases showed patchy shadow with pleuritis. One case presented sub-pleural curve line shadow at the posterior part of the right lower lung,and adhesion between the intestinal wall and intestinal mesentery in mass form in the abdomen by CT examination. Conclusion Patients suffering from AIDS (CD4T lymphocytes ＜50/μ L) with impaired immunity might be susceptible to complication of disseminated PM infection, which presents mainly damage of multiple organs and symptoms such as fever; enlargement of liver,spleen and lymph nodes, as well as specific skin maculopapular rashes. Imaging manifestations in the lungs were revealed as miliary nodular shadows and lattice-like shadows. Intensified abdominal CT might reveal presence of several enlarged postperitoneal lymph nodes and intestinal adhesion in shape of cakes.