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Objective:To evaluate the identification rate of separating gel or HB&L pretreatment methods of MALDI-TOF-MS, thereby to provide a new idea for the rapid and accurate identification of pathogens of bloodstream infections in daily clinic practice.Methods:A total of 149 alarmed positive blood culture samples of single bacterial infection by routine laboratory methods were collected between January to December 2020 from the Sixth Medical Center, Chinese PLA General Hospital. Samples were pretreated with the separation gel accelerating tube method or the HB&L microbial culture system, followed by direct MALDI-TOF MS bacterial identification, the identification rates of the two pretreatment methods were compared and results from the traditional method were used as the standard control.Results:Among the 149 positive blood culture samples, 47.0% (70/149) were gram-negative (G -) bacteria and 53.0% (79/149) were gram-positive (G +) bacteria. Identification rate of G -strain level was 78.6% (55/70) by serum separation gel coagulation tube method and 91.4% (64/70) by HB&L microbial culture system, the difference was statistically significant ( P=0.033). Identification rate of G +strain levels was 73.4% (58/79) by serum separation gel coagulation tube method and 87.3% (69/79) by HB&L microbial culture system, the difference was statistically significant ( P=0.028). For G -bacteria in the range of 3.000-2.300, the identification rate was 22.9% (16/70) by serum separation gel accelerating tube method and 38.6% (27/70) by the HB&L microbial culture system, the difference was statistically significant ( P=0.044). For G +bacteria in the range of 3.000-2.300, the identification rate was 19.0% (15/79) by serum separation gel accelerating tube method and 34.2% (27/79) by the HB&L microbial culture system, the difference was statistically significant ( P=0.031). Conclusion:The identification rate of HB&L microbial culture system is higher than that of serum separation gel coagulation tube method. Direct MALDI-TOF MS identification of pathogenic bacteria in positive blood culture samples after pretreatment is feasible in daily clinical practice.
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Objective To explore the impact of glucocorticoid on coagulation through administrating on rats with smoke inhalation.Methods Totally 150 male S-D rats were randomly (random number) divided into 5 groups:control group (ambient air inhalation),smoke group (smoke inhalation for 30 min),smoke+high dosage methyl prednisolone group(MP 40 mg/kg,intraperitoneal injection,s+HMP group),smoke+medium dosage MP (4 mg/kg) group (s+MMP group),smoke+low dosage MP (0.4 mg/kg) group (s+LMP group) (all n=30).Survival rates were calculated 24 h after smoke inhalation.Lung tissues were collected for histopathology and wet to dry (W/D) ratio.Arterial blood was collected for blood gas test.Coagulation factors in lung and plasma were tested.Results Survival rates of three MP groups were markedly improved compared with the smoke group (all P＜0.05),and was significantly higher in the medium dosage group(85.17％) than those in the low and high dosage groups (65.73％ and 60.07％,all P＜0.05).The W/D ratio and blood gas test were markedly improved in the high and medium groups (all P＜0.05).Tissue factor (TF) and thrombin-antithrombin complex (TAT-c) in bronchoalveolar lavage fluid (BALF) increased dramatically after SI (P＜0.01,P=0.005) with a remarkable drop of factor Ⅱ (F Ⅱ) (P=0.007),all of which were attenuated by MP with dosage dependence.The mRNA expression of TF increased dramatically after SI and recovered significantly with MP administration,while the expression of thrombomodulin (TM) recovered in the opposite direction with MP,all of which were in a dosage dependent manner.TF,fibrinogen (FIB),TAT-c increased significantly in plasma after smoke inhalation (P＜0.01,P=0.027,P=0.005).F Ⅷ ％ increased with MP administration and TF was raised by high dosage MP compared with the smoke group.FIB and TAT-c were decreased in all MP groups,which were significant higher in the high and middle dosage groups.The change of TM and endothelial cell protein C receptor (EPCR) in circulation were similar with FIB or TAT-c with or without MP.Protein C (PC％) and antithrombin (AT Ⅲ ％) dropped dramatically after SI,high and middle dosages of MP could restore the activity significantly,while low dosage would restore AT Ⅲ ％ but not PC％.Conclusions Glucocorticoid can significantly improve local and systemical coagulation disorder caused by smoke inhalation,and high-and medium-dosage hormones are effective.The regulation of hormones on the coagulation system is an important mechanism in the treatment of smoke inhalation induced lung injury.
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Objective To investigate the therapeutic effect of different doses of methylprednisolone (MP) in smoke inhalation-induced acute lung injury (SI-ALI).Methods Adult male Sprague-Dawley (SD) rats were divided into control group (group A,n = 6), smoke inhalation group (group B, smoke inhalation 30 minutes,n = 30) and smoke+MP 40, 4, 0.4 mg/kg intervention group (groups C, D, E; intraperitoneal injection of MP at 1 hour before smoke inhalation, n = 30) according to random number table method. The survival status of rats in each group was observed at 24 hours, and murine smoke inhalation induced trauma score (MSITS) according to the symptoms and signs of rats at 3 hours after smoke inhalation were scored. The blood of abdominal aorta of rats was collected. Then the rats were sacrificed to harvest bronchoalveolar lavage fluid (BALF) and lung tissue. The levels of interleukin (IL-6, IL-17a) in plasma and BALF were detected by enzyme linked immunosorbent assay (ELISA); the total number of white blood cells and the proportion of leukocytes or macrophages in BALF were calculated; the histopathological changes of lung were observed and the lung injury score was given; the expression of myeloperoxidase (MPO) and high mobility group protein B1 (HMGB1) in lung tissue were detected by Western Blot.Results The 24-hour survival rate of group B rats was 33.67％. The survivalrate of groups C, D and E (65.73％, 85.17％, 60.07％) were significantly higher than that of group B (allP < 0.05), and the survival rate of group D was significantly higher than that of groups C and E. Diffuse inflammatory cell infiltration, intra-alveolar hemorrhage and a large amount of edema fluid were seen in the lung tissue of group B; and the lung injury score was significantly higher than that of group A. Compared with group B, the lung injury in different doses of MP group were decreased to different degrees, while the lung injury scores in groups C and D were significantly decreased (3.31±1.37, 2.62±0.98 vs. 5.52±0.97, bothP < 0.01); correlation analysis showed that MSITS score was significantly and positively correlated with lung injury score (r = 0.862,P < 0.001). The levels of plasma inflammatory factors and BALF protein, inflammatory cells and inflammatory factors, and the expression of MPO, HMGB1 in group B were significantly higher than those in group A. Compared with group B, the levels of inflammatory factors in plasma, and protein content, inflammatory cells and inflammatory factors in BALF in different doses of MP group were decreased to different degrees, with significant differences in groups C and D [plasma: IL-17a (pg/L): 49.28±27.12, 36.57±16.52 vs. 191.79±88.21; IL-6 (ng/L): 206.47±109.96, 197.52±113.86 vs. 669.00±299.60; BALF: protein content (mg/L):892.0±164.5, 566.1±120.9 vs. 1838.0±145.8; white blood cell count (×109/L): 5.40±1.67, 2.81±1.20 vs. 9.02± 2.06; neutrophil ratio: 0.315±0.081, 0.273±0.080 vs. 0.590±0.096; IL-17a (ng/L): 22.63±8.62, 18.92±8.43 vs. 43.31±19.17; IL-6 (ng/L): 156.49±46.94, 123.66±64.91 vs. 253.43±80.03; allP< 0.01]; in addition, the expression of MPO and HMGB1 protein in lung tissues of MP groups with different doses were significantly decreased, the expression of MPO in group D was significantly lower than that in group E [MPO/β-actin (fold increase from group A):2.14±0.97 vs. 4.35±0.87,P < 0.01], the expression of HMGB1 in groups C and D were significantly lower than that in group E [HMGB1/β-actin (fold increase from group A): 1.77±0.73, 1.23±0.67 vs. 3.65±1.08, bothP < 0.05]. Conclusions MP can significantly improve the survival rate of SI-ALI rats and reduce the acute pulmonary and systemic inflammatory response. The MP effect of 4 mg/kg was better than 40 mg/kg and 0.4 mg/kg.
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Objective To explore the effects of hydrogen sulfide (H2 S) on the inflammatory reaction and the expression of single immunoglobin IL-1 receptor related protein (SIGIRR) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods Forty male SD rats were randomly divided into 4 groups (normal group, H2 S group, ALI group and ALI + H2 S group). The ALI rat model was established by LPS peritoneal injection. After LPS stimulation for 1 h, rats inhaled H2 S 80mg /m3 for 6h. Then, rats were sacrificed with a supraphysiological dose of pentobarbital sodium. The histological changes in the lung, the levels of TNF-α and IL-1β in serum and lung tissue homogenates, and the protein expression of SIGIRR in lung tissues were examined. Results Compared with the normal and H2 S groups, typical histological features of ALI were observed in ALI group, and the lung injury scores of ALI group were higher than those of the normal and H2 S groups (P ＜ 0. 05). Moreover, there were marked increases in the levels of TNF-α and IL-1β in serum and lung tissue homogenates after LPS injection. In contrast, inhalation of H2 S could attenuate lung pathological changes and inhibit the productions of TNF-α and IL-1β in serum and lung tissue homogenates (P ＜ 0. 05). Additionally, inhalation of H2 S could induce the protein expression of SIGIRR in rat lung tissues. Conclusion Inhalation of H2 S protected rats from LPS-induced ALI and its mechanisms were partially associated with inhibition of the productions of TNF-α and IL-1β and modulation of SIGIRR expression.
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Objective To explore the effect of curcumine on the inflammation and expression of single immunoglobin IL-1 receptor related protein in alveolar epithelial cells induced by lipopolysacharride (LPS).Methods The rat type Ⅱ alveolar epithelial cells were cultured in vitro,and cell activity was measured when stimulated with LPS and different doses of curcumin.The level of tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) in supematant was detected.Cells pretreated with curcumin (20 μmol/L),were stimulated with LPS (10 μg/mL).The nuclear protein and membrane protein was extracted to detect the expression level of nuclear transcription factor kappa B (NF-κB) and single immunoglobin IL-1 receptor related protein (SIGIRR).Results The cells activities were not affected by curcumin (5 ～30 pμmol/L) and LPS (10 μg/mL) (P ＜ 0.05).Curcumin (5 ～30 μ mol/L) significantly inhibited LPS-induced overpression of TNF-α and IL-6 (P ＜ 0.05).In 20 μ mol/L and 30 μ mol/L pretreatment groups,the inhibition of curcumin was most obvious,and there were no significant differences between the two groups (P ＜ 0.05).Curcumin (20 μ mol/L) significantly inhibited the expression level of phosphorylation of NF-κB p65 in cell nucleus,while up-regulated the expression of SIGIRR (P ＜ 0.05).Conclusion Curcumin inhibits the expression of inflammatory factor such as TNF-α and IL-6 as well as activation of NF-κB in alveolar epithelial cells induced by LPS.Up-regulating the expression level of negative regulatory molecules SIGIRR is one of the possible mechanism.
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Objective To observe the effects of seawater immersion at different temperatures on survival time and mortality and physiological state of non-anesthetized rats.Methods Totally 100 SD male rats(The abdominal cavity was implanted with a temperature sensor in advance)were randomly divided into five groups which were immersed in 20 ℃,17 ℃,15 ℃,13 ℃ and 10 ℃ seawater,respectively.Each group contains 20 rats.The changes of respiration,heart rate and muscle fibrillation within 2 hours were observed and the survival time and mortality of each group were counted in 24 hours.The decrease trend of intraperitoneal temperature in rats was analyzed retrospectively within 2 hours.Results Soaking for 10 minutes,the respiratory and heart rate of each group were significantly increased,but there was no significant difference among groups(P>0.05).The respiratory and heart rate decreased rapidly between 10 and 40 minutes,and the decline was slower relatively between 40 and 80 minutes.Soaking for 80 minutes,the respiration rate of rats among groups had significant difference(P0.05),hereafter the muscle fibrillation was maintained at a certain level.The mean survival time of 20 ℃ group,17 ℃ group,15 ℃ group,13 ℃ group and 10 ℃ group in 24 hours were(23.6±1.23)hours,(15.0±4.16)hours,(7.7±3.21)hours,(2.4±0.91)hours and (1.1±0.39)hours,respectively,and the survival curve of each group was statistically significant(P<0.05).The intraperitoneal temperature of rats showed a decline in the cliff,the lower the water temperature,the faster the descending.Soaking for 40 minutes,the difference of intraperitoneal temperature of each group was statistically significant(P<0.05).Conclusion The effects of seawater immersion at different temperatures on the physiological state and survival time and mortality of rats are significantly different.With the decrease of water temperature,the physiological state changes more obviously,the survival time is shorter and the mortality rate is higher.
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Objectives To explore the effect of the standardized residents training method on intern teaching in department of respiratory. Methods Sixty interns of Grade 2010 from the Second Military Medical School were randomly divided into 2 groups with 30 interns each. The traditional teaching method was adopted in control group, while the standardized residents training method was used in experiment group. When the rotating internship was finished in department of respiratory, the survey of satisfaction about teaching and the assessment of teaching effect were performed in two groups. The contents of examination included academic knowledge exam, clinical skills test, basic skills test and comprehensive quality assessment. Results There was no difference between two groups in academic knowledge exam (P>0.05). The total scores, clinical skills scores and basic skills scores of experiment group were higher than those of control group (P<0.05). And, higher satisfaction was ac-quired in experiment group(P<0.05). Conclusions Using the standardized residents training method can improve the effect of the intern teaching and teaching satisfaction and it can be widely applied in intern teaching.
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Objective To investigate the status of invasive fungal infection(IFI)associated with hematopathy,and evaluate drug resistance and risk factors of fungal infection.Methods 1 246 cases of infection occurred in patients in a hospital from 2006 to 2010 were analyzed retrospectively,pathogenic features and risk factors of IFI were ana-lyzed.Results There were 281 cases of fungal infection,and 162 fungal isolates were isolated,the main infection site was respiratory tract(134 isolates,82.72%).Four major Candida were Candida albicans ,Candida tropicalis , Candida glabrata ,and Candida krusei ;in 2006-2009,the main fungi were Candida albicans ,while in 2010,the majority were non-Candida albicans .The resistant rates of four isolated Candida to fluconazole and itraconazole were 5.15% and 4.41 % respectively,6 isolated Candida krusei strains were all resistant to both fluconazole and itraconazole,voriconazole-resistant strain was not found.The independent risk factors for fungal infection were dia-betes and duration time of agranulocytosis>14 days.Conclusion The proportion of infection caused by non-Candi-da albicans increased obviously,fluconazole-and itraconazole-resistant non-Candida albicans strains have emerged, comprehensive measures should be adopted to prevent IFI actively and treat patients early.
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<p><b>OBJECTIVE</b>To explore the value of ¹⁸F-FDG PET-CT in evaluating bronchial mucosa involvement in patients with saroidosis.</p><p><b>METHODS</b>A retrospective analysis was conducted among 6 sarcoidosis patients with and 14 patients without bronchial mucosa involvement to collect the data including the standard uptake value (SUVMax/Mean) of ¹⁸F-FDG, serum angiotensin converting enzyme (sACE), and proportion of lymphocytes and CD4⁺/CD8 ⁺ T lymphocyte ratio in bronchoalveolar lavage fluid (BALF).</p><p><b>RESULTS</b>The lung focal SUV(Max/Mean) was higher in patients with bronchial mucosa involvement than those without (7.04 ± 5.83/5.00 ± 4.69 vs 5.68 ± 3.66/3.82 ± 2.39), but such differences were not statistically significant (P=0.565/0.495). The SUV(Max/Mean) of the hilum of the lung and the mediastina lymph nodes were significantly higher in patients with bronchial mucosa involvement (13.28 ± 5.57/10.48 ± 4.43 vs 6.20 ± 1.77/4.52 ± 1.43, P=0.0003/0.0002; 13.84 ± 4.35/9.69 ± 2.74 vs 7.16 ± 2.52/5.28 ± 1.77, P=0.0004/0.0004). The level of sACE and CD4⁺/CD8 ⁺ T lymphocyte ratio in BALF were also significantly higher in patients with bronchial mucosa involvement (60.58 ± 16.3 vs 49.16 ± 13.3 IU/L, P=0.045; 7.30 ± 5.0 vs 2.90 ± 3.1, P=0.026). The proportion of lymphocytes in BALF was comparable between the patients with and without bronchial mucosa involvement (44.10 ± 10.3% vs 35.30 ± 12.5%, P=0.148).</p><p><b>CONCLUSIONS</b>For patients with saroidosis, ¹⁸F-FDG PET-CT is useful in evaluating bronchial mucosa involvement, which is one of the key features of active sarcoidosis.</p>
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Humanos , Bronquios , Patología , Líquido del Lavado Bronquioalveolar , Relación CD4-CD8 , Fluorodesoxiglucosa F18 , Ganglios Linfáticos , Patología , Peptidil-Dipeptidasa A , Sangre , Tomografía de Emisión de Positrones , Mucosa Respiratoria , Patología , Estudios Retrospectivos , Sarcoidosis , DiagnósticoRESUMEN
Objective To explore the diagnostic value of video bronchoscope-guide transbronchial needle aspiration(TBNA) combined with transbronchial lung biopsy(TBLB) on pulmonary sarcoidosis.Methods Twenty-two patients,definitely diagnosed as stage Ⅰ pulmonary sarcoidosis,were enrolled into the study and accepted TBNA and TBLB.Results The positive diagnostic rates of TBNA and TBLB were 63.6%(14/22) and 40.9%(9/22),but the rate increased to 90.9%(20/22) when the two methods were used together,which showed significant differences(x2=12.24,4.66,P＜0.01 or ＜0.05).Conclusion TBNA combined TBLB test is a safety method with high diagnostic accuracy for pulmonary sarcoidosis.
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0.05). After ventilation with 6ml/kg tidal volumes, the TNF-? in alveolar lavage fluid (1.58?0.56pg/ml) was significantly lower in b1 group compared with that in control group (1.73?0.81pg/ml, P
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Objective: To observe the therapeutical effect of intermittent positive pressure ventilation (IPPV) on pulmonary edema after seawater drowning in rabbits.Methods:Fourteen seawater lavage induced pulmonary edema New Zealand rabbits models were assigned to receive IPPV treatment.Blood gas analysis, pulmodynamics, hemodynamics status and serum tumor necrosis factor(TNF ?) were monitored at various time points.After 3 h ventilation rabbits of each group were killed to gain lung tissues for pathology examination, and lung lavage fluid was analyzed to count leukocytes.Results:PaO 2 significantly increased after the use of ventilation in pulmonary edema rabbits models ( P