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Tyrosine-decahydrofluorene derivatives are a class of hybrid compounds that integrate the properties of polyketides and nonribosomal peptides. These compounds feature a [6.5.6] tricarbocyclic core and a para-cyclophane ether moiety in their structures and exhibit anti-tumor and anti-microbial activities. In this study, we constructed the biosynthetic pathway of xenoacremones from Xenoacremonium sinensis ML-31 in the Aspergillus nidulans host, resulting in the identification of four novel tyrosine-decahydrofluorene analogs, xenoacremones I-L (1-4), along with two known analogs, xenoacremones A and B. Remarkably, compounds 3 and 4 contained a 12-membered para-cyclophane ring system, which is unprecedented among tyrosine-decahydrofluorene analogs in X. sinensis. The successful reconstruction of the biosynthetic pathway and the discovery of novel analogs demonstrate the utility of heterologous expression strategy for the generation of structurally diverse natural products with potential biological activities.
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Aspergillus nidulans/metabolism , Biological Products/metabolism , Polyketides/metabolism , Peptides/metabolism , Biosynthetic Pathways , Multigene FamilyABSTRACT
Objective:To investigate the effect of activation of microglia in prefrontal cortex on long-term spatial memory in post-stroke depression mice.Methods:Forty-eight male C57BL/6 mice were divided into sham operation group, stroke group, post-stroke depression group and depression group according to the random number table method with 12 in each group, and 36 mice were divided into solvent group, enrofloxacin group and minocycline group according to the random number table method with 12 in each group.Middle cerebral artery occlusion (MCAO) was use to establish the stroke model, and forced swimming was used to establish the depression model.The post-stroke depression model mice were received MCAO first and then received forced swimming on the 4th day after stroke to establish the model.Mice in enrofloxacin group and minocycline group were treated with enrofloxacin and minocycline injection once a clay for 14 days from the 5th day after stroke, respectively.Forced swimming test and sugar water preference test were used to evaluate the depression of mice in each group, Morris water maze test was used to detect the spatial memory function of mice in each group, and Nissl staining and immunofluorescence staining were used to detect the neuronal function and the number and type of microglia activation.The expression of inflammatory cytokines IL-6 and IL-1β were detected by Western blot.GraphPad Prism 8.0.1 statistical software was used for statistical analysis.The single factor variance analysis was used to compare the difference among multiple groups, and pairwise comparison was performed with SNK- q test. Results:(1) There were statistically significant differences in depression, learning and memory, neuron damage, activation of microglia, inflammatory factors and other indicators in sham operation group, stroke group, post-stroke depression group and depression group ( F=43.58-255.70, all P<0.05). Compared with stroke group, post-stroke depression group had longer floating immobility time ((222.70±29.12) s, (79.25±46.78) s, P<0.05), the preference rate of sugar water was significantly lower ( (49.44±6.19) %, (84.49±4.73) %, P<0.05), and the average value of platform approach after correction was higher((125.00±9.95) mm, (96.79±12.57) mm, P<0.05), Nissl bodies expression was lower ((53.50±15.78) cells /mm 2, (85.67±17.52) cells /mm 2, P<0.05), NeuN positive expression rate was lower ((29.78±3.70) %, (45.73±4.51) %, P<0.05), the percent of M1 microglia expression was significantly higher ((75.55±8.84) %, (58.19±5.69) %, P<0.05), the percent of M2 microglia expression was lower ((43.46±5.11)%, (57.14±5.40)%, P<0.05), and the expression levels of IL-6 ((1.14±0.03), (0.94±0.05), P<0.05) and IL-1β((1.17±0.03), (0.56±0.04), P<0.05) were significantly higher.(2) Depression, learning and memory, neuron injury, activation of microglia, inflammatory factors and other indicators of mice in solvent group, enrofloxacin group and minocycline group were significantly different ( F=7.13-94.35, all P<0.05). Compared with enrofloxacin group, mice in minocycline group had shorter floating immobility time ((169.30±13.04) s, (224.30±22.60) s, P<0.05) and higher sugar water preference rate ((62.81±7.75) %, (47.71±8.11) %, P<0.05), the mean value of platform approach estimation after water maze correction was lower ((97.66±14.56) mm, (120.20±12.08) mm, P<0.05), and the expression level of Nissl bodies was higher ((80.17±10.55) cells /mm 2, (52.00±8.94) cells /mm 2, P<0.05), NeuN expression rate was high ((45.04±3.62) %, (28.88±4.50) %, P<0.05), Iba-1 expression was lower ((97.33±10.67) cells/mm 2, (112.50±6.54)cells/mm 2, P<0.05), the percent of M1 microglia expression was lower ((54.43±5.22) %, (73.82±6.88) %, P<0.05), and the percent of M2 microglia expression was significantly higher ((51.86±6.22) %, (36.30±5.72) %, P<0.05). The expression levels of IL-6 ((0.75±0.06), (1.21±0.07), P<0.05) and IL-1β ((0.61±0.06) (1.09±0.09), P<0.05) were lower. Conclusion:The long-term spatial memory impairment of post-stroke depression mice is aggravated, which is related to the neuron damage caused by increased activation of M1 microglia in PFC area.Inhibition of M1 microglia by minocycline can effectively improve the spatial memory ability of mice.
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Objective:To investigate patients' compliance of endoscopic surveillance after endoscopic submucosal dissection (ESD) and the influencing factors.Methods:This study was a cross-sectional survey. The data of patients who underwent ESD in Xijing Digestive Hospital of Air Force Military Medical University from January 2014 to December 2015 were collected through questionnaire and telephone call. The questionnaire survey was conducted from May 1, 2019 to July 31, 2020. Patients' compliance for endoscopic surveillance and influencing factors were evaluated. Logistic regression model was used for multivariate analysis to determine independent risk factors affecting compliance of endoscopic surveillance.Results:A total of 413 questionnaires were collected, of which 331 (80.1%) patients were well complied with endoscopic surveillance, and 82 (19.9%) were not. Univariate analysis showed that gender ( χ2=4.68, P=0.034), lesion type ( χ2=10.10, P=0.002), educational background ( χ2=4.00, P=0.049), disposable income ( χ2=7.00, P=0.009), employment ( χ2=11.29, P=0.004) and medical insurance ( χ2=13.59, P=0.001) affected patients' endoscopic surveillance compliance. Logistic regression analysis indicated that patients with submucosal lesions were less likely to be compliant than patients with mucosal lesions with the same type of medical insurance ( P<0.001, OR=0.383, 95% CI: 0.227-0.644); and with the same lesion type, patients with urban medical insurance were more likely to be compliant than those with rural cooperative medical insurance ( P<0.001, OR=2.938, 95% CI: 1.696-5.090). Conclusion:The endoscopic surveillance compliance for post-ESD patients is related to the lesion type, the type of medical insurance, and the awareness of the disease. More attention should be paid to those with submucosal lesions and rural cooperative medical insurance due to their poor compliance. And it is necessary to improve health education on the disease for better surveillance compliance.
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Tyrosine-decahydrofluorene derivatives feature a fused [6.5.6] tricarbocyclic core and a 13-membered
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Objective:To explore a scoring system of preoperative evaluation for high-risk gastric stromal tumors.Methods:A total of 275 consecutive patients with gastric stromal tumors of diameter ≤5 cm who underwent surgical or endoscopic resection at Xijing Hospital between 2008 and 2018 were retrospectively enrolled in the study. Multivariate logistic regression analysis was performed to investigate independent risk factors for high-risk gastric stromal tumors.Weighted points was proportionally assigned based on β regression coefficient value to establish the scoring system. The receiver operator characteristic (ROC) curve was plotted, the scoring system as the test variable, and the area under curve (AUC) was calculated.Results:Multivariate analysis revealed tumor size>1.75 cm ( OR=6.474, 95% CI: 2.335-17.948), irregular tumor shape ( OR=3.548, 95% CI: 1.745-7.216) and mucosal ulceration ( OR=2.412, 95% CI: 1.154-5.041) were independent risk factors for high-risk gastric stromal tumors of diameter ≤5 cm ( P<0.05). These risk factors for high-risk gastric stromal tumors were weighted with value: one point for mucosal ulceration, two points for irregular tumor shape and three points for tumor size>1.75 cm. The AUC value of the scoring system was 0.781, of which the cut-off value was 4. According to the cut-off value, 0-3 points was categorized as the low risk group and 4-6 points as the high risk group for gastric stromal tumors. The incidence of high-risk gastric stromal tumors of the low risk group and the high risk group were 13.3% (26/196) and 48.1% (38/79), respectively, with significant difference ( χ2=38.266, P<0.001). Conclusion:Tumor size larger than 1.75 cm, irregular tumor shape and mucosal ulceration can be applied to establish a preoperative scoring system to predict high-risk gastric stromal tumors of diameter ≤5 cm.
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The discovery of hydroxylases in the anticancer drug taxol biosynthesis pathway is a hotspot and difficulty in current research. In this study, a new hydroxylase gene TcCYP725A22 (GenBank accession number: MF448646.1) was used to construct a sub-cellular localization vector pCAMIBA1303-TcCYP725A22-EGFP to get the transient expression in onion epidermal cells. Laser confocal microscopy revealed that the protein encoded by this gene was localized in the cell membrane. Furthermore, the recombinant plant expression plasmid pBI121-TcCYP725A22 was constructed. After transient transformation to the Taxus chinensis mediated by Agrobacterium tumefaciens LBA4404, qRT-PCR and LC-MS were utilized to analyze the effects of TcCYP725A22 overexpression on the synthesis of taxol. The results showed that, in the TcCYP725A22 overexpressed cell line, expression levels of most defined hydroxylase genes for taxol biosynthesis were increased, and the yield of taxanes were also increased. It was concluded that the hydroxylase gene TcCYP725A22 is likely involved in the biosynthetic pathway of taxol.
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Biosynthetic Pathways , Mixed Function Oxygenases , Paclitaxel , Taxoids , TaxusABSTRACT
Objective@#To analyze the epidemiological characteristics of human brucellosis and trace back source of infection of human brucellosis in Hunan province during 2010-2018, and provide evidence for the prevention and control of human brucellosis.@*Methods@#The surveillance data of human brucellosis in Hunan during 2010-2018 were analyzed with software Excel 2016 and ArcGIS 10.5, the epidemic characteristics were described using cases number, constituent ratio and rate. The conventional biotype methods were used for the identification of Brucella species, UTS-PCR was applied to further confirm the results from conventional biotype detections, then six virulence genes of two clinical Brucella strains were detected by PCR assay. Cluster analysis of two Brucella strains were performed with Multiple locus variable-number tandem repeat analysis (MLVA) for the investigation of the infection source of human brucellosis.@*Results@#From 2010 to 2018, a total of 728 human brucellosis cases were reported in Hunan with the annual incidence rate of 0.12/100 000. The incidence rate was 2.50/100 000 in Chenzhou and 1.90/100 000 in Yongzhou, higher than those in other areas. The number of counties reporting cases increased from 5 in 2010 to 69 in 2018. Most cases were reported in age group 45-54 years, accounting for 38.32% (279/728). The cases in farmers accounted for 59.07% (430/728) of the total. The male to female ratio of the cases was 2.75 ∶ 1. The reported case number was highest during May-July, accounting for 45.33% (330/728). The incidence was high in summer and autumn, and the peak was in May. The conventional identification showed that two strains were all Brucella melitensis biovar 1, consistent with UTS-PCR amplification results. Six virulence genes were found in two isolated strains, suggesting that the Brucella melitensis strains in this study had strong virulence. MLVA results confirmed that two strains detected in Hunan had complete identical MLVA-16 genotype with strains isolated from goat and camel in Inner Mongolia Autonomous Region, indicating that there was molecular epidemiology relationship between these strains and the source of infection were originated from Inner Mongolia.@*Conclusions@#The epidemic of human brucellosis in Hunan is becoming serious, and disease has spread to general population and non-epidemic areas. Two Brucella melitensis strains detected in Hunan were originated from Inner Mongolia. The quarantine and inspection in animal transportation should be strengthened to prevent human outbreaks of brucellosis.
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Objective@#To investigate the history, risk factors for prognosis of malignant biliary stricture (MBS) patients receiving conservative therapy after endoscopic retrograde cholangiopancreatography(ERCP) and to set up a predictive model for overall survival (OS).@*Methods@#MBS patients who underwent ERCP and conservative therapy in Xijing Hospital and PLA No.451 Hospital from January 2009 to December 2013 were enrolled to the present study. Predictive factors associated with OS were identified in the training cohort by stepwise multivariate Cox regression analysis. A predictive model was then developed and externally validated in the validation cohort.@*Results@#Between January 2009 and December 2013, 152 and 149 patients were eligible to the training and validation cohort respectively. In the training cohort, tumors were mainly originated from bile duct (33.6%), pancreas (23.5%) or ampulla (20.4%). 76.3% (116/152) patients died during the observation period. The median OS for the training population was 5.0 months (3.9-6.2 months). CA19-9≥1 000 U/mL, non-ampulla tumor, metastasis, pre-ERCP total bilirubin≥7 mg/dL and hilar stricture were identified as independent predictive factors of poor OS (all P<0.05). Based on these factors, the COMTH predictive model was developed. The median OS of patients with COMTH>8 in the training and validation cohorts were both 3.0 months, which were significantly shorter than those with COMTH≤8 (10.0 and 6.9 months in the training and validation cohorts respectively, both P<0.001).@*Conclusion@#The prognosis of MBS patients undergoing ERCP is poor. The survival chance of patients with COMTH>8 is even more dismal.
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Objective To observe the levels of peripheral blood cytokines interferon-γ (IFN-γ)and interleukin-4 (IL-4) in patients with acute and chronic brucellosis before and during treatment,and to understand the differences of two immunocytokines in acute and chronic stage of brucellosis,and the effect of antibacterial therapy on these two cytokines to provide immunological basis for clinical evaluation of the therapeutic effect of brucellosis.Methods Research subjects were 36 pre-treatment acute brucellosis and 36 pre-treatment chronic brucellosis and 36 dur-treatment acute brucellosis and 36 dur-treatment chronic brucellosis,which were selected from Ulanqab Center for Endemic Disease Prevention and Control of Jining City with 25 local healthy persons as healthy controls.The levels of IFN-γ and IL-4 in serum were measured by enzyme-linked immunosorbent assay (ELISA) in patients with acute and chronic brucellosis and control group before and during treatment.Parameters of IFN-γ and IL-4 were analyzed with One-Way ANOVA analysis in pre-treatment and dur-treatment acute brucellosis,chronic brucellosis and control groups.Results The means of IFN-γ [(462.79 ± 47.94),(431.92 ± 40.39),(280.50 ± 40.48) ng/L] and IL-4 [(606.11 ± 51.86),(550.66 ± 51.56),(383.24 ± 53.98) ng/L] were significantly different in the three groups before treatment (F =141.84,139.28,P ＜ 0.05);Compared to control group,the levels of IFN-γ and IL-4 in acute brucellosis and chronic brucellosis were significantly increased before treatment (P ＜ 0.05).The levels of IFN-γand IL-4 in the acute brucellosis were significantly increased compared to those of chronic brucellosis before treatment (P ＜ 0.05).After about ten days antibiotic therapy,the means of IFN-γ [(356.05 ± 43.75),(368.61 ± 35.69),(280.50 ± 40.48) ng/L] and IL-4 [(487.31 ± 51.59),(496.73 ± 48.70),(383.24 ± 53.98) ng/L] were significantly different in the three groups (F =39.57,41.99,P ＜ 0.05).Compared to control group,the levels of IFN-γ and IL-4 in acute brucellosis and chronic brucellosis were significantly increased during treatment (P ＜ 0.05).The levels of IFN-γand IL-4 in the acute brucellosis were not significantly different compared to those of chronic brucellosis during treatment (P ＞ 0.05).Conclusion Different immunological characteristics of cytokines in peripheral blood of patients with acute and chronic brucellosis before treatment have affected the therapeutic effect and clinical outcomes.
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Objective To explore risk factors of influencing operating efficiency of endoscopic submucosal dissection (ESD) for gastric mucosal lesions. Methods The data of 304 cases with gastric mucosal lesion undergoing ESD in Xijing Hospital of Digestive Disease from April 2009 to February 2017 were retrospectively analyzed. The procedure time and complete resection rate ( R0 resection rate ) were regarded as indicators of ESD efficiency. Risk factors influencing procedure time and R0 resection rate were analyzed using Chi-square test and logistic regression. Results Using median procedure time of 45 min as the cutoff value, Chi-square test showed that specimen size ( P=0. 000) , lesion location ( P=0. 001) , and pathological type ( P=0. 003) affected the operation time. Further logistic regression analysis indicated that specimen size (≥40 mm/<40 mm, P<0. 001, OR=3. 748, 95％CI: 2. 247-6. 254) and lesion location (upper or middle 1/3 of stomach/lower 1/3 of stomach, P=0. 001, OR=2. 180, 95％CI: 1. 318-3. 606) were independent risk factors of procedure time. Using R0 resection as outcome measure, neither single nor multiple parameter analysis was statistically significant. Conclusion Specimen size and lesion location are independent risk factors influencing efficiency of gastric mucosal ESD, and could be possibly used to estimate the procedure time of ESD.
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Objective To compare the safety and efficacy of endoscopic submucosal tunnel dissection (ESTD) and endoscopic submucosal dissection ( ESD) for treatment of esophageal superficial neoplasms. Methods A retrospective study was performed on data of patients with esophageal superficial neoplasms who were treated by conventional ESD or ESTD at Xijing Hospital of Digestive Diseases between January 2014 and December 2016. The procedure time, pathology, rate of en bloc resection and curative resection, and adverse events were compared between the two groups. Results A total of 113 consecutive patients were collected, including 49 undergoing ESTD and 64 undergoing ESD. ESTD had a shorter procedure time than ESD method [38. 0 min(21. 4-71. 0 min) VS 46. 5 min(32. 5-117. 5 min), P＝0. 008],and the dissection speed of ESTD was faster than that of ESD[0. 42 cm2/min(0. 22-0. 59 cm2/min) VS 0. 34 cm2/min(0. 20-0. 42 cm2/min), P＝0. 000]. There were no statistical differences in the en bloc resection rate ( 100. 0% VS 100. 0%, P＝1. 000) or the curative resection rate (98. 0% VS 93. 8%, P＝0. 386).There were no statistical differences on adverse event rates including post-procedure bleeding, perforation, fever, and thoracalgia.ESTD group showed a lower rate of muscular injury (20. 4% VS 39. 1%, P＝0. 041) and intra-procedure bleeding (18. 4% VS 37. 5%, P＝0. 036). A multivariate regression analysis for procedure time showed that ESTD method ( OR＝ 2. 801, 95%CI: 1. 116-7. 031, P＝0. 028) and lesion area ＜9 cm2(OR＝5. 049, 95%CI: 2. 088-12. 208, P＝0. 000) were associated with a shorter procedure time.Conclusion ESTD is safe and effective for treatment of esophageal superficial neoplasms. It can shorten operative time, improve dissection speed, and reduce intra-procedure muscular layer injury and bleeding.
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Objective To evaluate the endoscopic features of colorectal sessile serrated adenoma/polyp(SSA/P). Methods The data of 109 cases of SSA/P and 218 cases of polyps randomly selected in Xijing Hospital from January 2014 to December 2016 were collected. The endoscopic features of SSA/P and polyps were compared, and the risk factors of occurrence and cancerization of SSA/P were analyzed. Results The mean age of patients in the SSA/P group was older than that of polyps group(P=0.011).The distribution of lesions was no significant difference between the two groups(P=0.092). The gross type of SSA/P was mainly type Ⅰ and Ⅱ, while polyps were more in type Ⅰ(P=0.036). According to the pit pattern of Kudo,type Ⅱ was more in SSA/P,but types Ⅰ and Ⅱ were more in polyps(P=0.004). For capillary pattern comparison,type Ⅱ was more in SSA/P,but type Ⅰ was more in polyps(P≤0.000 1). For surface morphological features comparison,the SSA/P group was more likely to be observed the mucous cap(P=0.002)and blood vessel thickening(P=0. 004). On pathologic diagnosis, the SSA/P group was more susceptible to atypia and carcinogenesis(P = 0.001). Higher microvascular morphological classification,being mucous cap,and blood vessel thickening were risk factors of atypia and carcinogenesis of SSA/P. Conclusion There were significant differences between SSA/P and polyps on lesion location,pit pattern,capillary pattern,surface structure characteristics,and risk factors of atypia and carcinogenesis.
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Objective To investigate the HOOF genotyping characteristics of 83 Brucella (B.)melitensis strains isolated in Ulanqab of Inner Mongolia Autonomous Region from 2012 to 2015.Methods A total of 83 B.melitensis strains were detected by convention identification and AMOS-PCR,then HOOF protocol with eight VNTR locus were used for the genotyping of the strains,and the allelic diversity of each VNTR locus and the discriminatory power of VNTR typing of HOOF were assessed by Hunter-Gaston Discriminatory index.BioNumerics 5.0 was used for phylogenetic analysis and constructing dendrogram.Results All of the isolates were identified as B.melitensis strains by two identification methods.The complete eight VNTR locus had higher polymophism and diversity index was 0.998;and diversity index of six locus (1,2 and 4-7) were ≥0.678,discriminatory power of HOOF was mainly from this six higher diversity index locus.The 83 B.melitensis strains were classified into eight clusters and 76 genotypes,6 shared genotypes included 13 isolates,indicating that these brucellosis cases had epidemiological link,the other 70 strains had distinct genotypes,indicating that these cases had no epidemiological link.Conclusions The epidemic of human brucellosis in Ulanqab was characterized by local and sporadic outbreaks.Cross infection was related with the transfer of the sources of infection.
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Objective To get knowledge of the molecular epidemiological characteristics of human derived Brucella isolated in Hohhot,and to provide experimental basis in guiding prevention and treatment of Brucella infection.Methods Twenty-seven Brucella isolates derived from patients in Affiliated Hospital of Inner Mongolian Medical University from 2013 to 2015 were identified by routine bacteriological methods and molecular methods.Multiple-locus variable number tandem repeats analysis (MLVA-16) was used to detect molecular typing and do cluster analysis.Sixteen virulent genes were detected and analyzed by polymerase chain reaction (PCR).Results Twenty-seven Brucella isolates were identified as Brucella melitensis (B.melitensis) by routine bacteriological methods and PCR.Out of them,six isolates were B.melitensis biovar 1,and twenty-one isolates were B.melitensis biovar 3.MLVA-16 analysis showed that seven genotypes were obtained from nine Brucella isolates,which showed significant difference in variable number of tandem repeats,which suggested that they originated from sporadic outbreak.Moreover,two isolates were clustered into the same clade,which suggested they were epidemiologically correlated and may be derived from the same origin.Sixteen virulent genes were detected in all of the twenty-seven isolates.Conclusions Brucella isolates from patients in Hohhot are mainly B.melitensis biovar 3 and B.melitensis biovar 1,and the distribution profile of multiple virulence genes is similar.Some isolates have showed epidemic correlation,and the epidemic mechanism should be further explored.
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Paeonol has inhibitory effect on a variety of tumor cells, whereas cadherin is a kind of glycoprotein that is associated with the occurrence and development of different tumor.In this study, the interactions of paeonol and E-cadherin have been investigated by fluorescence spectrometry and atomic force microscopy (AFM).Fluorescence spectrometry results revealed that the addition of paeonol significantly quenched the fluorescence of E-cadherin.Based on the results of the quenching constant, it was inferred that the interaction of paeonol and E-cadherin was a static quenching process.The thermodynamic parameters ΔH and ΔS were calculated to be -4.3×10.5 J/mol and -1.3×10.3 J/(mol·K), respectively, which proved the involvement of weak interactive forces such as hydrogen bond van der Waals force.AFM results revealed that cadherin molecules were assembled into the long-chain structure.The addition of paeonol could significantly disrupt these assembling structures into short chains, which could be ascribed to the damage of the interdigitation model from the adjacent cadherin molecules.All these results reveal that cadherin is an important target of paeonol to modulate its activity.
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Objective To explore the change and value of interleukin (IL)-2,IL-6 and tumor necrosis factor (TNF)-α in the serum samples of patients with brucellosis.Methods The levels of serum IL-2,IL-6 and TNF-α were measured using the enzyme linked immunosorbent assay (ELISA) method in 155 patients with brucellosis in different clinical stages (including 69 cases in acute,34 cases in subacute and 52 cases in chronic periods) and 50 healthy controls.IL-2 was used to represent the helper T cell (Th)-type 1 cytokines,IL-6 represent Th2 type cytokines,and the Th1/Th2 ratio in different clinical stages was compared.Results The expression level of serum IL-2 in patients with brucellosis was different (ng/L:acute:10.15 ± 2.01;subacute:9.53 ± 1.68;chronic:6.76± 1.31;control:47.25 ± 5.68),the differences were statistically significant (F =74.921,P ＜ 0.05).The expression levels of serum IL-2 in acute,subacute and chronic periods of brucellosis patients were significantly lower than that of healthy controls (all P ＜ 0.05).The expression levels of serum IL-2 in acute and subacute periods of brucellosis patients were higher than that of chronic period patients (all P ＜ 0.05).The expression of IL-6 and TNF-α in serum of patients in different clinical stages were different (ng/L:acute:615.22± 341.07,802.55 ± 479.53;subacute:478.45 ± 105.33,680.21 ± 366.95;chronic:306.37 ± 96.12,455.36 ± 176.27;control:121.45 ± 30.16,87.51 ± 24.03),the differences were statistically significant (F =57.692,63.210,all P ＜ 0.05).The levels of serum IL-6 and TNF-o were significantly higher in patients with brucellosis in acute,subacute and chronic clinical stages than that of healthy controls (all P ＜ 0.05).Serum IL-6 and TNF-α levels in acute period of brucellosis patients were higher than those of subacute and chronic periods patients (all P ＜ 0.05);the levels of serum IL-6 and TNF-α in subacute period of brucellosis patients were also significantly higher than that of chronic period patients (all P ＜ 0.05).The Th1/Th2 ratio was different (acute:0.02 ± 0.00;subacute:0.02 ± 0.00;chronic:0.23 ± 0.02;control:0.41 ± 0.06) in clinical patients with brucellosis,the differences were statistically significant (F =17.843,P ＜ 0.05),the value of Th1/Th2 in acute and subacute period patients were lower than that of chronic period (all P ＜ 0.05).Conclusion IL-2,IL-6 and TNF-α may play an important role in the pathogenesis of brucellosis,and their dynamic changes can reflect the progression and outcome of brucellosis.
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Objective We compared peripheral blood nucleotide-binding oligomerization domain-leucinerich repeats containing pyrin domain 3 (NLRP3),cysteinyl aspartate specific proteinase-1 (Caspase-1),interleukin-1β (IL-1β) and IL-18 between acute and chronic brucellosis patients before treatment and revealed their immune characteristics,to find targets for immune intervention of brucellosis.Methods From March to April 2016,42 acute and 42 chronic brucellosis patients were selected from Ulanqab Center for Endemic Disease Prevention and Control as the research subjects,and 20 local healthy persons were selected as healthy control.Brucellosis were diagnosed according to the "Diagnostic Criteria of Brucellosis".Enzyme-linked immunesorbent assay (ELISA) method was used to determine serum levels of NLRP3,Caspase-1,IL-1β and IL-18.Results The expression levels of NLRP3,Caspase-1,IL-1β and IL-18 [(1 264.40 ± 424.74),(1 350.67 ± 468.93),(192.96 ± 61.52),(162.74 ±54.23),(172.44 ± 60.56),(120.10 ± 61.52),(47.23 ± 13.79),(46.68 ± 14.72),(27.71-± 8.71),(202.23 ± 65.24),(169.19 ± 54.33),(108.62 ± 41.39) ng/L] were compared in acute,chronic and control groups,the differences were statistically significant (F =61.96,6.26,16.68,18.31,P ＜ 0.01).Compared to control group,the levels of NLRP3,Caspase-1,IL-1β and IL-18 in acute and chronic groups were significantly increased (P ＜ 0.05);compared to chronic group,the levels of IL-18 in acute group were significantly increased (P ＜ 0.05),and NLRP3,Caspase-1 and IL-1 β levels were not statistically different (P ＞ 0.05).Conclusions This study has showed that the expression of NLRP3 inflammasome in innate immunity is not significantly different between acute brucellosis and chronic brucellosis.The difference of IL-18 levels between acute and chronic brucellosis may affect their immune response.
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Objective To establish genotyping methods for rapid identification of Brucella melitensis (B.melitensis) biovar 1,2 and 3 and to verify these method.Methods Single nucleotide polymorphism of RpoB gene and tandem repeat sequence (TRS) Bru42 of standard reference strain 16M were used to design primers,then the RpoB-PCR and TRS-PCR method were established for identification of B.melitensis standard reference strains,these two methods were used to identify clinical isolates of B.melitensis and compared with the conventional methods.Results The results of B.melitensis standard reference strains (biotype 1,2,3) identified by RpoB-PCR and TRS-PCR were consistent with those of the conventional identification methods.Totally 50 clinical isolates [including B.melitensis biovar 1 (17),2 (3) and 3 (30)] were identified as RpoB-2 genotype,only one B.melitensis biovar 1 strain was identified as RpoB-3 genotype.Genotype identification results of standard reference strains and clinical isolates with the same biotype were not exactly the same.Fothermore,TRS-PCR experiment displayed that 51 clinical isolates were all genotype 2 of B.melitensis (genotype TRS-2).Conclusions There is no clear relationship between biovars and genotypes within B.melitensis,and significant difference exists between B.melitensis standard reference strains and clinical isolates within RpoB gene.Bru42 can not be used for genotyping clinical isolates of B.melitensis.
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Objective To establish genotyping methods for rapid identification of Brucella melitensis (B.melitensis) biovar 1,2 and 3 and to verify these method.Methods Single nucleotide polymorphism of RpoB gene and tandem repeat sequence (TRS) Bru42 of standard reference strain 16M were used to design primers,then the RpoB-PCR and TRS-PCR method were established for identification of B.melitensis standard reference strains,these two methods were used to identify clinical isolates of B.melitensis and compared with the conventional methods.Results The results of B.melitensis standard reference strains (biotype 1,2,3) identified by RpoB-PCR and TRS-PCR were consistent with those of the conventional identification methods.Totally 50 clinical isolates [including B.melitensis biovar 1 (17),2 (3) and 3 (30)] were identified as RpoB-2 genotype,only one B.melitensis biovar 1 strain was identified as RpoB-3 genotype.Genotype identification results of standard reference strains and clinical isolates with the same biotype were not exactly the same.Fothermore,TRS-PCR experiment displayed that 51 clinical isolates were all genotype 2 of B.melitensis (genotype TRS-2).Conclusions There is no clear relationship between biovars and genotypes within B.melitensis,and significant difference exists between B.melitensis standard reference strains and clinical isolates within RpoB gene.Bru42 can not be used for genotyping clinical isolates of B.melitensis.
ABSTRACT
Objective To design a pressure detection and control system in the medical compartment for bio-agent casualty emergency evacuation and en route treatment during responses to biological warfare or terrorist attack.Methods An instrument and a pressure transmitter were used for detection,display and alarm of positive and negative pressures in the medical compartment,and pulse width modulation was involved in to regulate fan speed and then to control the pressure inthe compartment.Results The system proved its ability to evacuate bio-agent casualty and fulminating infectious patient with no pollution to the environment.Conclusion The system realizes on-line monitoring of the filter absorber in the medical compartment,enhances the efficiency and reliability of casualty care and thus gains high military benefits.