Automatic Identification of MALDI-TOF MS Database Using Classical Bordetella Species Isolates.

Objective: To evaluate and expand the automatic identification and clustering of clinical Bordetella species by MALDI-TOF MS. Methods: Twenty-eight field isolated strains, identified by whole-gene sequencing analysis, were analyzed by MALDI-TOF MS, and the spectra obtained were used to replenish the internal database of the manufacturer. To evaluate and expand the robustness of the database, MALDI-TOF MS identified 91 clinical isolates (except those used for implementation). A distance tree based on mass spectrometry data is constructed to confirm similarity and clusters of each clinical Bordetella species by using the MALDI Biotyper 3.1 software. Results: In this research, when we used the implemented Bruker Daltonics database in our laboratory, 91 clinical isolates were identified at the genus level (100%) and 93.4% were identified at the species level (85/91). We performed proteomics analysis and divided these 91 isolates into cluster I (2.2%) and cluster II (97.8%). The largest group is cluster II (n = 89 isolates), which has been divided into two subclusters. Trees created by analyzing the protein mass spectra of the three species of the clinical isolates reflected their classification. Conclusion: MALDI-TOF MS may present an attractive alternative to automatically confirm and cluster the fastidious bacteria difficult to culture. Extension of identification of the MALDI-TOF MS database is viably fast, more efficient, and alternative to conventional methods in confirming the classical Bordetella species. This strategy could promote the epidemiological and taxonomic research of this important pathogen.

Epidemiological and clinical characteristics of bacteremic brucellosis.

OBJECTIVE: To retrospectively investigate the epidemiological features, clinical manifestations and laboratory characteristics of bacteremic brucellosis. METHODS: Brucellosis patients admitted to our clinic from January 2015 to December 2017 were included in the study. Patient electronic medical records were reviewed for epidemiological features, clinical manifestations, and laboratory findings. RESULTS: A total of 132 brucellosis patients were analyzed (64 cases with bacteremic brucellosis and 68 cases with nonbacteremic brucellosis). The median duration from exposure to onset of symptoms was 6.9 weeks (range: 1 day to 32 weeks) and 21.9 weeks (range: 1-76 weeks) in patients with bacteremic and nonbacteremic brucellosis, respectively. More bacteremic than nonbacteremic patients presented with fever and chills. Arthritis was observed in 34 (25.8%) patients, and was more commonly observed in nonbacteremic patients. Using C-reactive protein (CRP) and procalcitonin (PCT) as serological markers, the areas under the receiving operating characteristic curves were 0.64 [95% confidence interval (CI): 0.54-0.73] and 0.61 (95% CI: 0.51-0.70), respectively, for distinguishing bacteremic from non-bacteremic brucellosis. CONCLUSION: Fever and chills were frequently observed in bacteremic brucellosis patients, whereas arthritis was more common in nonbacteremic brucellosis patients. Serum CRP and PCT can be used as potential serological markers for diagnosing bacteremic brucellosis.

Genetic association between mannose-binding lectin polymorphisms and viral hepatitis: a meta-analysis.

Some previous genetic association studies have tried to investigate potential associations between mannose-binding lectin (MBL) polymorphisms and viral hepatitis. However, the results of those studies were not consistent. Therefore, we performed the current meta-analysis to explore associations between MBL polymorphisms and viral hepatitis in a large pooled population. A systematic literature research of PubMed, Web of Science, Embase and CNKI was performed to identify eligible studies for pooled analyses. We used Review Manager version 5.3.3 to conduct statistical analyses. In total, 27 studies were included for analysis (4840 cases and 5729 controls). The pooled analyses showed that MBL promoter (-211C/G, dominant model: P = 0.0002, I2 = 40%; over-dominant model: P = 0.0001, I2 = 22%) and exon 1 (codon 52, 54 and 57, dominant model: P = 0.04, I2 = 49%; allele model: P = 0.01, I2 = 48%) polymorphisms were both significantly associated with viral hepatitis in the overall population. Further subgroup analyses revealed similarly significant findings for MBL promoter polymorphism in HBV and HCV, but no positive results were detected in subgroup analyses for MBL exon 1 polymorphism. These results suggested that MBL promoter and exon 1 polymorphisms could be used to identify individuals at higher susceptibility to HBV and HCV.

[Induction of anti-hepatocellular carcinoma immunity by Hyper-IL-6 gene in vivo].

AIM: To prepare the mouse hepatocellular carcinoma (HCC) cells with the stable expression of Hyper-IL-6 and explore the possibility of inducing active anti-HCC immune response by Hyper-IL-6 gene. METHODS: Hyper-IL-6 gene was transfected into mouse HCC cells MM45T.Li using Lipofectamine(TM);2000. G418-resistant clones named MM45T-HIL-6 were selected and detected for the expression of Hyper-IL-6 gene by RT-PCR and ELISA. Mouse HCC cells transfected with pEGFP-C1, named MM45T-mock, were prepared as controls. Tumor models were established by injecting subcutaneously 5×10(5); cells of MM45T.Li, MM45T- mock and MM45T-HIL-6 on the right anterior limb of BALB/c mouse, respectively. In vivo experiments were performed to observe the tumorigenicity of MM45T.Li, MM45T-mock and MM45T-HIL-6. The levels of CD4(+); and CD8(+); T cells in mouse peripheral blood were detected by flow cytometry (FCM). RESULTS: RT-PCR and ELISA showed that Hyper-IL-6 gene was expressed in the MM45T-HIL-6 cells, but not in the control cells. We observed that the tumorigenicity of MM45T-HIL-6 decreased compared with control cells after they were inoculated subcutaneously into mice. FCM results indicated that the levels of CD4(+); and CD8(+); T cells in the peripheral blood significantly increased in the mice inoculated with MM45T-HIL-6 compared with the ones inoculated with MM45T.Li and MM45T-mock cells (P<0.05). CONCLUSION: The mouse HCC cells with the stable expression of Hyper-IL-6 can induce active anti-HCC immune response after inoculated subcutaneously into mice.

[Construction and expression of fusion gene encoding Hyper-IL-6 in mammalian cells].

AIM: To construct and express fusion gene encoding Hyper-IL-6 in mammalian cells. METHODS: Using geneSOEing method, human sIL-6R cDNA was fused with IL-6 cDNA by a linker rich in glycine through PCR technique and cloned into pIRES(2)-EGFP. The constructed plasmid was transfected into mammalian cells. The expression of fusion gene encoding Hyper-IL-6 was detected by GFP and RT-PCR and immunocytochemistry. RESULTS: The recombinant plasmid pIRES-HIL-6 was successfully constructed and expressed in mammalian cells. CONCLUSION: The success in construction and expression of human Hyper-IL-6 makes it possible to further study its biological functions.

[The effect of artificial support system on serum cytokine in chronic severe hepatitis].

OBJECTIVE: To explore the changes of cytokines including TNFalpha, TGFbeta1 and nitrogen monoxide, and endotoxin in the serum of chronic severe hepatitis after the treatment of ALSS, and to evaluate further the value of ALSS in the treatment of chronic severe hepatitis. METHODS: Forty two patients were screened. The changes of TNFalpha, TGFbeta1, nitrogen monoxide and endotoxin were detected respectively. The relationship between the cytokines and the severity and prognosis were further analyzed. RESULTS: ALSS was effective to decrease the serum concentration of cytokines. TNFalpha dropped from (481.57+/-229.33) pg/ml to (156.46+/-78.12) pg/ml (P < 0.05). TGFbeta1 from (44.09+/-31.73) ng/ml to (27.77+/-23.28) ng/ml (P < 0.01), endotoxin from (1.05+/-0.37) Eu/ml to (0.28+/-0.22) Eu/ml (P < 0.001). NO from (71.15+/-33.09) micromol/L to (58.11+/-29.30) micromol/L (P < 0.001). Before the therapy endotoxin was related with TNFalpha and total bilirubin, while after the therapy, NO was related with protime and aminonemia. CONCLUSION: High level of endotoxin and nitrogen monoxide in serum plays an important role in hepatocyte damage of chronic severe hepatitis. The changes of serum endotoxin TNFalpha, TGFbeta1 and nitrogen monoxide level in patients with chronic severe hepatitis can be used to judge the severity and prognosis of severe hepatitis. ALSS is a reliable hepatic support device for chronic severe hepatitis