Objective To analyze epidemiological characteristics of brucellosis in Xinjiang Production and Construction Corps and to provide evidences for control and prevention of the disease.Methods Retrospective study method was used to collect the epidemic characteristics of cases and serological surveillance of brucellosis data in Xinjiang Production and Construction Corps from 2013 to 2017,the time,region,population distribution characteristics and laboratory test results of the disease were analyzed.Results In 2013-2017,the Xinjiang Production and Construction Corps reported a total of 3 287 cases of brucellosis,with an average annual incidence of 24.02/100 000,and no death case was reported.During the period,the incidence was 21.11/100 000,29.73/100 000,26.68/100 000,20.10/100 000 and 19.23/100 000,which showed a decline trend in general (linear x2 =23.67,P ＜ 0.01).The largest cumulative numbers of reported cases of the Corps were in five divisions,they were the Eighth Division with 599 cases,the Sixth Division with 470 cases,the Fourth Division with 366 cases,the Tenth Division with 320 cases and the Ninth Division with 313 cases,and they accounted for 62.91％ (2 068/3 287) of the total number of cases in the whole Corps.Brucellosis occurred in each month of the year,the number of cases reported from March to August accounted for 72.59％ (2 386/3 287),the peak incidence occurred in May and June,showing obvious seasonal fluctuation.Of all reported cases,74.78％ (2 458/3 287) were men,and 25.22％ (829/3 287) were women,38.97％ (1 281/3 287) were at the ages of 40-49 years,farmer was the highest occupation which accounted for 47.52％ (1 562/3 287).Totally 24 120 blood samples were collected from high-risk occupation during the period and 457 samples were positive for Brucella antibody,with an average positive rate of 1.89％,of the positive cases,52.30％ (239/457) had symptoms of brucellosis.Conclusions On the whole,the epidemic of brucellosis in Xinjiang Production and Construction Corps is declining year by year.But as the epidemic slowly expands,all departments should strengthen joint prevention and control measures,control and eliminate the source of infection.At the same time,we should strengthen the propaganda,education and behavior intervention for high-risk occupational groups.

Objective To investigate the roles of Dectin-1 in phagocytosis of Candida albicans (C.albicans) by macrophage-like cells derived from a human acute monocytic leukemia cell line THP-1.Methods THP-1 macrophage-like cells served as the target cells,and were transfected with small interfering RNA (siRNA) targeting Dectin-1 to down-regulate the expression of Dectin-1 receptor (siRNA-Dectin-1 group).THP-1 macrophage-like cells transfected with nonsense siRNA (siRNA-NC) served as a negative control group.After transfection,the THP-1 macrophage-like cells in the above 2 groups were cocultured with heat-killed C.albicans separately.And then,fluorescence microscopy was performed to count THP-1 macrophage-like cells phagocytosing C.albicans,and flow cytometry was used to determine the mean fluorescence intensity (MFI) of dihydrorhodamine (DHR)-123 fluorescent cells.Statistical analysis was done by one-way analysis of variance (ANOVA) and t test with the SPSS19.0 software.Results After transfection with siRNA-Dectin-1,the mRNA and protein expression of Dectin-1 significantly decreased in THP-1 macrophage-like cells (t =26.163,P ＜ 0.001).After 1-,2-,4-hour co-culture of THP-1 macrophagelike cells with C.albicans,fluorescence microscopy showed that the phagocytosis rates of C.albicans by THP -1 macrophage-like cells were significantly lower in the siRNA-Dectin-1 group than in the negative control group (17.5％ vs.22.1％,18.6％ vs.24.3％,39.2％ vs.59.1％,respectively,all P ＜ 0.05),so were the percentage of THP-1 macrophage-like cells phagocytosing more than 3 C.albicans cells (2.2％ vs.4.7％,2.5％ vs.5.4％,5.1％ vs.8.3％,respectively,all P ＜ 0.05).After 30-minute,1-,2-and 4-hour co-culture of THP-1 macrophage-like cells with DHR-123-labelled C.albicans,flow cytometry showed that the MFI of C.albicans-phagocytosing cells was significantly lower in the siRNA-Dectin-1 group than in the negative control group (36.8 vs.45.7,54.3 vs.62.4,72.1 vs.84.9,93.6 vs.116.7,respectively,all P ＜ 0.05).Conclusion Dectin-1 receptor plays an important role in the phagocytosis of C.albicans by macrophages.

Objective To summarize the efficacy of extracorporeal membrane oxygenation (ECMO)utilization in Emergency Department (ED),as well as the establishment of emergency ECMO team.Methods A retrospective analysis was carried out in 16 patients treated with ECMO between April 2015 to December 2016 in ED.The clinical data including demographics,diagnosis,initiating ECMO timing,place of ECMO establishment,intubation approaches,duration of ECMO,complications and outcomes were collected and analyzed.Results Eight patients were successfully weaned from ECMO,and 7 of them survived to discharge from hospital.The duration of ECMO support was 4 to 384 hours.The emergency ECMO team was set up.Conclusions Emergency medical team can successfully operate the ECMO process.The emergency medical team-initiated ECMO can provide effectively adjuvant measures to support patients with respiratory failure,circulatory failure and cardiac arrest.

Objective To evaluate effects of the yeast form of Sporothrix schenckii on activation of p38 mitogen-activated protein kinase (p38MAPK) and expression of interleukin-6 (IL-6) in macrophagelike THP-1 cells,which were differentiated from the human acute monocytic leukemia cell line THP-1.Methods THP-1 macrophage-like cells were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii at a concentration of 2 × 106 colony-forming units (CFU)/ml (yeast form group),100 mg/L curdlan (curdlan group) and RPMI 1640 medium (blank control group) respectively.Real-time fluorescence-based quantitative PCR was performed to measure the mRNA expression of IL-6 in THP-1 macrophage-like cells in the above 3 groups after 3-and 6-hour treatment separately,and enzyme-linked immunosorbent assay (ELISA) to detect the level of IL-6 in the culture supernatant of THP-1 macrophagelike cells after 24-hour treatment.Western blot analysis was conducted to determine the protein expression of p38MAPK and phosphorylated p38MAPK (p-p38MAPK) in the above 3 groups after 30-and 60-minute treatment separately.Other THP-1 macrophage-like cells were pretreated with 100 nmol/L dexamcthasonc (a p38MAPK inhibitor) for 30 minutes,and then were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii,curdlan and RPMI 1640 medium respectively,and changes in the level of pp38MAPK and mRNA expression of IL-6 were also detected.Statistical analysis was carried out with SPSS19.0 software by using one-way or multi-way analysis of variance and least significant difference (LSD) test.Results Significant differences in the mRNA expression of IL-6 in THP-1 macrophage-like cells were observed among the yeast form group,curdlan group and blank control group (F =5 552.22,P ＜0.001) after 3-hour treatment (56.81 ± 7.36,26.69 ± 1.22 and 0.97 ± 0.05,respectively) and 6-hour treatment (378.03 ± 16.67,276.24 ± 39.13 and 1.02 ± 0.04,respectively).Additionally,the yeast form group showed significantly higher mRNA expression of IL-6 after 6-hour treatment than that after 3-hour treatment (q =16.74,P ＜ 0.001).After 24-hour treatment,the level of IL-6 in the culture supernatant of THP-1 macrophage-like cells also significantly differed among the yeast form group,curdlan group and blank control group (59.96 ± 18.16 pg/L,91.01 ± 17.27 pg/L,5.50 ± 2.30 pg/L,respectively;F =26.62,P ＜ 0.01),and was significantly higher in the yeast form group than in the blank control group (P ＜ 0.01).After 30-and 60-minute treatment,the protein expression of p-p38MAPK was significantly higher in the yeast form group than in the blank control group (both P ＜ 0.01).Moreover,the mRNA expression of IL-6 (4.46 ± 1.03 vs.493.52 ± 113.87,P ＜ 0.001) and protein expression of p-p38MAPK (2.29 ± 0.37 vs.4.55 ±0.46,q =10.81,P ＜ 0.01) were both significantly lower in the yeast form group with dexamethasone pretreatment than in that without dexamethasone pretreatment.Conclusion In vitro treatment with the yeast form of Sporothrix schenckii can enhance the expression of IL-6 in human THP-1 macrophage-like cells by activating the p38MAPK signaling pathway.

Objective:To investigate the biological characteristics of monoclonal antibodies against avian influenza virus (AIV).Methods:Monoclonal antibodies (mAbs) against AIV H5N1 were prepared and it′s characteristics were identified including subtype,titer,hemagglutination inhibition activity and cross-reactivity with other influenza viruses.Besides,Western blot and immuno-histochemical staining methods were conducted to test the combination of antibodies and antigen ( H5N1 ) and human normal tissues.Results:Immunohistochemical analysis showed that 2 mAbs (H5-32 and H5-35) cross-reacted with human tissues kidney and pancreas respectively.Conclusion:These data indicated that there have some association between the AIV H 5N1 with human tissues, which may provide reference for the study on avian influenza virus infection and pathogenicity .

Objective To investigate the effects of Candida albicans on the expression of tumor necrosis factor-α(TNF-α)and activation of the intracellular signaling molecule p38 mitogen-activated protein kinase(p38MAPK)in a human acute monocytic leukemia cell line THP-1. Methods Some THP-1 cells were divided into several groups in vitro: two C. albicans groups treated with 105 CFU/ml and 106 CFU/ml heat-killed C. albicans respectively, a lipopolysaccharide (LPS)group treated with 100 μg/L LPS, a blank control group treated with RPMI 1640 medium, two dexamethasone-inhibited groups pretreated with 40 μg/L dexamethasone for 30 minutes followed by treatment with 106 CFU/ml heat-killed C. albicans and LPS respectively. After treatment for 1, 3 and 6 hours, real-time fluorescence-based quantitative PCR was performed to measure TNF-α mRNA expression in THP-1 cells in the above groups. Enzyme-linked immunosorbent assay(ELISA)was conducted to determine the level of TNF-α protein in the supernatant of THP-1 cells treated with 106 CFU/ml heat-killed C. albicans, 100 μg/L LPS or RPMI 1640 medium(blank control group)for 24 hours. Western blot was performed to measure the protein expression of p38MAPK and phosphorylated p38MAPK in THP-1 cells after treatment with 106 CFU/ml heat-killed C. albicans or RPMI 1640 medium (blank control group)for 30 and 60 minutes. Statistical analysis was carried out by using two-way analysis of variance, one-way analysis of variance and the least significant difference(LSD)-t test. Results Significant differences were observed in the mRNA expression level of TNF-α among the C. albicans groups, LPS group and blank control group (F = 110.98, P < 0.001). The mRNA expression level of TNF-α in THP-1 cells increased over time in a time-dependent manner after C. albicans treatment, with significant differences among different time points (F = 701.680, P < 0.001). Compared with the blank control group, both 106-CFU/ml C. albicans group and LPS group showed a significant increase in TNF-α protein expression (6385.70 ± 533.99 ng/L and 3212.06 ± 353.00 ng/L vs. 147.10 ± 0.53 ng/L, P < 0.001 and 0.005, respectively). An obvious increase was observed in the expression level of phosphorylated p38MAPK protein, but no significant changes were noted in that of p38MAPK protein, in THP-1 cells treated with 106 CFU/ml C. albicans for 30 and 60 minutes compared with the blank control group. The mRNA expression level of TNF-α significantly decreased in dexamethasone-pretreated 106-CFU/ml C. albicans group and LPS group compared with those without dexamethasone pretreatment(3.77 ± 0.62 vs. 208.50 ± 10.50, 6.20 ± 1.93 vs. 161.35 ± 1.65, both P < 0.001). Conclusions Heat-killed C. albicans can induce the activation of p38MAPK in and secretion of TNF-α by human THP-1 cells, which then participate in the innate immune response against C. albicans.

Objective To investigate whether human neutrophils kill Candida albicans through recognition of insoluble β-glucan in cell walls of C.albicans (CalG) by dectin-1,a C-type lectin receptor.Methods Neutrophils were obtained from peripheral blood of healthy human subjects and cultured in vitro.Real-time PCR was carried out to quantify the mRNA expressions of dectin-1 and Toll-like receptor 2 (TLR2) in neutrophils challenged with CaIG of 100 mg/L for 1,6,and 24 hours.A Fluoro hydrogen peroxide (H2O2) detection kit was used to determine H2O2 levels in some neutrophils exposed to CaIG (100 mg/L) for 15 minutes,2 hours,6 hours,as well as in some neutrophils preincubated with laminarin (a dectin-1 inhibitor) of 100 mg/L and 50 mg/L for 30 minutes followed by challenge with CaIG of 100 mg/L for 2 hours.Colony forming units (CFUs) were counted after the incubation of C.albicans with neutrophils pretreated with laminarin of 100 mg/L and 50 mg/L for 30 minutes.Results The relative mRNA expression level of dectin-1 was 2.8195 + 0.1669,5.4859 + 0.7244 and 3.6041 + 0.5372 in neutrophils challenged with CaIG for 1,6 and 24 hours,respectively,significantly higher than that in unchallenged neutrophils at these corresponding time points (all P ＜ 0.01).The level of H2O2 was (64.55 + 15.67),(290.34 + 30.56),and (208.54 ± 26.88) μ mol/L respectively in neutrophils treated with CaIG for 15 minutes,2 hours,and 6 hours respectively,compared to (22.05 ± 3.99) μmol/L in untreated neutrophils (all P ＜ 0.01).The pretreatment with laminarin of 100 and 50 mg/L attenuated the release of H2O2 in CaIG-treated neutrophils by 73％ ((80.45 + 22.41) μ mol/L,P＜ 0.01) and 45％ ((130.42 + 44.55) μmol/L,P＜ 0.01),respectively,compared with neutrophils treated with CaIG only.The fungicidal activity of neutrophils against C.albicans was also significantly inhibited by pretreatment with laminarin of 50 and 100 mg/L (both P＜ 0.01).Conclusions Dectin-1 may be involved in the secretion of H2O2 as well as killing of C.albicans by human neutrophils,which may provide a preliminary evidence for adoptive transfer of neutrophils as an approach to the treatment of systemic C.albicans infection.

Objective To evaluate the in vitro activity of seven imidazole antifungals against clinical isolates of common Candida species.Methods According to the Clinical and Laboratory Standards Institute (CLSI) microdilution method M27-A3,the in vitro activity of luliconazole,ketoconazole,miconazole,econazole,clotrimazole,sertaconazole and bifonazole was determined among 183 clinical isolates belonging to 5 species of Candida.Results The minimal inhibitory concentration range was 0.03-8 (geometric mean:0.067) mg/L for ketoconazole,0.03-16 (geometric mean:0.071 ) mg/L for miconazole,0.03-8 (geometric mean:0.207) mg/L for econazole,0.03-8 (geometric mean:0.061 ) mg/L for clotrimazole,0.03-16 (geometric mean:0.187) mg/L for sertaconazole and 0.03 -＞16 (geometric mean:1.050) mg/L for bifonazole. Luliconazole exhibited a superior activity against the 5 species of Candida in vitro,with the MIC range being 0.03-8 mg/L,geometric mean MIC 0.087 mg/L,MIC50 0.06 mg/L and MIC90 0.5 mg/L,respectively.However,some Candida isolates were identified to be relatively insensitive to these tested antifungals,including luliconazole.Conclusion All the tested imidazole antifungals,except for bifonazole,show an excellent activity against Candida species in vitro,but there exist a few Candida strains with relative insensitivity.

A 19-year-old man was admitted to the hospital for erythema and nodules on the face and postauricular region for 6 years. Microscopic examination of lesion scrapings revealed brown septate hyphae. A restricted, velvety and black colony grew on Sabouraud's dextrose agar. Slide culture on potato dextrose agar plate showed flask-shaped phialides at the apex of or around the hyphae with clear collarettes and flaring apex,mucilage-encapsuled, round to oval, semi-endogenous phialosporae accumulating at the apex of the phialides,giving a flower-like appearance. Anti-fungal susceptibility test showed that the fungus was sensitive to itraconazole, terbinafine and amphotericin B, but resistant to fluconazole. Sequence analysis of the ITS1-ITS4 region revealed a 98% consistency with the reference sequence of ITS1-ITS4 of Phialophora verrucosa. On the basis of above findings, the patient was diagnosed with cutaneous phaeohyphomycosis. Clinical improvement was seen after treatment with oral itraconazole (400 mg/d).

A 16-year-old woman presented plaques on the left auricle and face over a period of 3 years. Fungal culture grew black-grey or dust velvety colony on Sabouraud's dextrose agar plate. A slide culture on potato dextrose agar plate showed conidiophores which were unbranched or occasionally loosely branched. The conidia were sympodial, zero- to two- septate, with rounded apices and truncated bases. The optimum growth temperature was 26℃ - 30℃. The fungus had the ability to liquefy glutin and hydrolyze starch. Anti-fungal susceptibility test showed the fungus was susceptible to itraconazole, terbinafine and amphoterecin B, but resistant to fluconazole. Cutaneous biopsy specimens revealed brown hyphae and budding yeast cells. The sequence of internal transcribed spacer (ITS) 1-ITS4 region of the isolate rDNA was assessed and compared against the Genebank databases. A 99% consistence was observed in the ITS sequence between clinical isolate and reference strain of Veronaea botryose Ciferri et Momtemartini. Based on the above findings, the mold was identified as Veronaea botryose Ciferri et Momtemartini. The lesions gradually subsided after 8-month treatment with oral itraconazole of 100 mg twice daily.