Objective In this article we evaluated the sensitivities and specificities of real-time PCR assay for diagnosis of human brucellosis.Methods The species selectivity and specificity of real-time PCR were evaluated by direct amplification of a 169 bp portion of bcsp31 gene from 15 Brucella strains and 41 non-Brucella strains.According to the monitoring results of 2012 Harbin brucellosis,17 brucellosis patients and 30 health people were selected to collect their serum samples for assessing the sensitivity of real-time PCR,and additional 30 nonbrucellosis patients serum samples were as controls.Results The species selectivity and specificity of our realtime PCR method were evaluated by using genomic DNA from 15 Brucella strains and 41 non-Brucella strains.There were 11 sera with positive amplification signals among the 17 culture-proven brucellosis patients,the sensitivity was 64.7％(11/17).Whereas,the results of sera from the 60 control patients were all negative,corresponding to a specificity of 100.0％.Conclusion The results indicate that real-time PCR is well suitable for confirmation of brucellosis cases.

ObjectiveTo study the clinical effect of improved decortication in treatment of tuberculous empyema as well as the safety of operation.MethodsEighty-two cases who diagnosed as tuberculous empyema by pathology and operation from January 2007 to September 2010 were selected including improved decortication 44 cases,total empyema decortication 28 cases,thoracoplasty 10 cases.The operation time,peri-operative bleeding,postoperative complication and lung function recovery after 6 months operation were followed-up.ResultsAll the patients were in good recovery and there was no death in the operation for 1 year.The peri-operative bleeding in improved decortication was less than that in total empyema decortication and thoracoplasty [(56.23 ± 15.56) ml vs. (78.65 ± 23.14) and (66.92 ± 19.83) ml],and there was significant difference among them(P＜ 0.01 ).There was no postoperative complication in improved decortication,but 2 cases (7.1％,2/28) of partial lung recruitment maneuvers in total empyema decortication,1 case ( 10.0％,1/10) of extensive staxis with selective surgery in thoracoplasty.The proportion of forced vital capacity (FVC),forced expiratory volume in one second (FEV1),peak expiratory flow (PEF) turning to normal after 6 months operation in improved decortication [95.5％(42/44),93.2％(41/44),97.7％(43/44)]were higher than those in total empyema decortication[ 75.0％ (21/28 ),78.6％ (22/28),85.7％ (24/28) ]and thoracoplasty [ 80.0％ (8/10),90.0％ (9/10),80.0％ ( 8/10) ],and there was significant difference among them (P ＜0.01).There was no significant difference in the operation time among them(P ＞0.05).ConclusionImproved decortication has remarkable superiority in curing tuberculous empyema especially it is small in traumatic with less blood loss,fewer postoperative complications,lung function recovery rapidly after operation.

Acute Disease ; Animals ; Charcoal ; therapeutic use ; Enema ; Milk ; Paraquat ; toxicity

Purpose: Our research aimed to investigate the influence of miR-103a-3p on the growth and apoptosis of colorectal cancer (CRC) cells. Materials and Methods: Bioinformatics was employed to analyze differentially expressed microRNAs and predict target genes. qRT-PCR was applied to detect the expression of miR-103a-3p in CRC and normal cells. HCT116 and Caco-2 were chosen, and miR-103a-3p mimics, miR-103a-3p inhibitor, as well as specific siRNAs targeting GREM2, were constructed. We subsequently evaluated alternations in cell proliferation, cell cycle and cell cycle regulators, apoptosis, and related proteins (Bcl-2 and Bax) by CCK-8 testing, Western blotting, luciferase reporter, colony formation, and Annexin V-FITC/PI. Possible binding sites for miR-103a-3p on the 3'UTR of GREM2 were checked with luciferase assay, and the impact of GREM2 on miR-103a-3p activity was also validated with above biological function testing. Additionally, the effect of miR-103a-3p knockdown in CRC cells and the molecular mechanism of miR-103a-3p targeting GREM2 were also studied. Results: Bioinformatics analysis revealed that miR-103a-3p expression increased remarkably in CRC, and targeted regulatory correlation existed between miR-103a-3p and GREM2. MiR-103a-3p inhibitor significantly impeded proliferative capacity and caused cell cycle arrest, as well as apoptosis, in HCT116 and Caco-2 cells. Consistent with this finding, overexpression of GREM2 showed similar effects to miR-103a-3p inhibition. Moreover, we demonstrated that miR-103a-3p connected target GREM2 and GREM2 knockdown reversed the effects of miR-103a-3p inhibitor on HCT116 and Caco-2 cell proliferation, cell cycle, and apoptosis. Further study showed that miR-103a-3p targeting GREM2 appeared to affect CRC progression via the transforming growth factor-β pathway. Conclusion MiR-103a-3p could augment CRC progression by targeting GREM2 and that miR-103a-3p/GREM2 could be potential novel targets for CRC therapy.

Objective:To investigate the effects and mechanisms of low-dose hydrocortisone on myocardial injury in early septic shock rats.Methods:Seventy-two healthy male Sprague-Dawley (SD) rats were divided into Sham group, lipopolysaccharide (LPS) model group (LPS group) and low dose hydrocortisone intervention group (LD group) according to the random number table method, with 24 rats in each group. The rat model of septic shock was produced by intravenous injection of LPS at 20 mg/kg. Sham group was injected with an equal amount of physiological saline. The LD group was injected 5 mg/kg of hydrocortisone via right femoral vein after model establishment. Sham group and LPS group were injected with an equal amount of physiological saline. Blood pressure and heart rate (HR) of rats in each group were continuously monitored. In each group, 8 rats were sacrificed for arterial blood gas analysis at 0, 3 and 6 hours after model establishment, and the level of plasma N-terminal B-type brain natriuretic peptide precursor (NT-proBNP) was detected by enzyme linked immunosorbent assay (ELISA). The expressions of cleaved-caspase-3 and nuclear factor-κB p65 (NF-κB p65) were detected by Western Blot. Myocardial tissue was harvested 6 hours after model establishment, the histopathological changes were observed by hematoxylin eosin (HE) staining, and the apoptosis rate of myocardial cells was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL).Results:After LPS injection, mean arterial pressure (MAP) decreased significantly at 1 hour, then gradually increased, and was significantly higher than Sham group at 6 hours. There was no significant change in HR, and the difference was not statistically significant compared with Sham group. Blood lactic acid (Lac), base excess (BE), plasma NT-proBNP level, myocardial tissue caspase-3 and NF-κB p65 expression increased with the extension of time, all reach the peak in 6 hours, and significantly higher than Sham group. After early treatment with low-dose hydrocortisone in septic shock, MAP showed an increasing trend and Lac, BE decreased slowly. At 6 hours, MAP, Lac and BE were significantly lower than those in the LPS group [MAP (mmHg, 1 mmHg = 0.133 kPa): 98.6±7.5 vs. 106.1±8.5, Lac (mmol/L): 1.29±0.08 vs. 2.42±0.37, BE (mmol/L): 4.45±0.57 vs. 8.18±1.03, all P < 0.05]. The level of plasma NT-proBNP, and the expressions of caspase-3 and NF-κB p65 in myocardial tissue were significantly lower than those in LPS group at 3 hours and 6 hours after low-dose hydrocortisol treatment [NT-proBNP (ng/L): 2 740.56±97.31 vs. 4 425.60±743.32 at 3 hours, 2 638.81±205.12 vs. 4 993.01±373.78 at 6 hours; caspase-3/GAPDH: 0.567±0.045 vs. 0.841±0.162 at 3 hours, 0.496±0.071 vs. 1.116±0.172 at 6 hours; NF-κB p65/GAPDH: 0.852±0.734 vs. 1.232±0.115 at 3 hours, 0.783±0.047 vs. 1.383±0.215 at 6 hours, all P < 0.05]. HE staining results showed that myocardial cells in the LPS group were broken and inflammatory cells infiltrated. The myocardial histopathological changes in LD group were significantly less than those in LPS group. TUNEL staining showed that the apoptosis of myocardial cells in LPS group increased, and the apoptosis rate was significantly higher than that in Sham group [(82.41±1.57)% vs. (5.77±0.69)%, P < 0.05]. The apoptosis rate in LD group was significantly lower than that in LPS group [(27.82±1.77)% vs. (82.41±1.57)%, P < 0.05]. Conclusion:Low-dose hydrocortisone plays a protective role in the myocardial injury of early septic shock, and its mechanism may be related to the inhibition of caspase-3 and NF-κB p65 expression, the reduction of apoptosis rate and myocardial inhibition.