In vitro synergistic antibacterial activity of isochlorogenic acid combined with antibiotics against polymicrobial biofilms formed by Pseudomonas aeruginosa and Aspergillus fumigatus / 中华微生物学和免疫学杂志

Objective:To study the synergistic antibacterial effect of honeysuckle active components isochlorogenic acid combined with voriconazole+ ceftazidime on the early polymicrobial biofilms of Pseudomonas aeruginosa and Aspergillus fumigatus. Methods:The polymicrobial biofilm models in the early stages were established in vitro under static condition and then treated with isochlorogenic acid at different concentrations (250, 500, 1 000 μg/ml) and antibiotics (1 μg/ml of voriconazole+ 16 μg/ml of ceftazidime) alone or in combination for 24 h in vitro. Morphological changes in the polymicrobial biofilm structure were observed under scanning electron microscope (SEM). Crystal violet staining assay was used for polymicrobial biofilm quantitation. The total metabolic activity of Pseudomonas aeruginosa and Aspergillus fumigatus in formed polymicrobial biofilms were calculated with 2, 3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay. Results:Under the SEM, it was observed that isochlorogenic acid could destroy the structure of early polymicrobial biofilms and reduce the extracellular matrix. The inhibitory effect of isochlorogenic acid combined with antibiotics against the polymicrobial biofilms was stronger than that of the antibiotics alone, and the amount of biofilms on the carrier was significantly reduced. Crystal violet staining assay showed that the total amount of biofilms in isochlorogenic acid (250, 500, 1 000 μg/ml) groups was significantly different from that in isochlorogenic acid (250, 500, 1 000 μg/ml) combined with antibiotics groups, antibiotics group and blank control group (all P<0.05). Moreover, there were significant differences between different concentration groups ( P<0.05). XTT assay suggested that the metabolic activity in the isochlorogenic acid combined with antibiotics group was significantly lower than that in the blank group and isochlorogenic acid group (the inhibition rate reached 50%). Conclusions:Isochlorogenic acid could not kill the Aspergillus fumigatus and Pseudomonas aeruginosa in polymicrobial biofilms in vitro, but exhibit destructive effect on the polymicrobial biofilms in the early stages. Isochlorogenic acid combined with voriconazole and ceftazidime had inhibitory effects against the early stages of polymicrobial biofilms of Aspergillus fumigatus and Pseudomonas aeruginosa in a concentration-dependent manner and was stronger than voriconazole combined with ceftazidime.

Effects of cinnamaldehyde on aspergillus fumigatus biofilm in vitro / 重庆医学

Objective To study the treatment effect of cinnamaldehyde on Aspergillus fumigatus biofilm(BF) in vitro .Meth‐ods The models of A .fumigatus BF were established in vitro ;the minimum inhibitory concentration(MIC) on A .fumigatus was measured .The crystal violet assay and scanning electron microscopy were employed to determined the treatment effect of A .fumiga‐tus biofilm under varying concentrations of cinnamaldehyde .Results BF models were established successfully in vitro .MIC value of A .fumigatus of cinnamaldehyde was 256 μg/mL ;The biofilm biomass in serially increasing concentrations of cinnamaldehyde(1 MIC ,1/2 MIC ,1/4 MIC)were 0 .81 ± 0 .11 ,1 .13 ± 0 .18 and 1 .59 ± 0 .11 respectively .Compared to untreated control group(2 .18 ± 0 .15) ,difference achieved statistical significance(P<0 .05) .SEM studies revealed the deformity of three‐dimensional structures of biofilms treated with sub‐MICs of cinnamaldehyde .Conclusion Cinnamaldehyde has significant antifungal activity against Aspergil‐lus fumigatus ,sub‐MICs could disrupt the mature biofilm in vitro .

In vitro efficacy of the combination of Cinnamaldehyde and Voriconazole on Aspergillus fumigatus isolates / 天津医药

Objective To evaluate the in vitro antifungal activity of Cinnamaldehyde in combination with Voriconazole (VRC) against clinically isolated Aspergillus fumigatus strains. Methods According to the Clinical and Laboratory Stan?dards Institute (CLSI) M38-A2 document,the minimal inhibitory concentrations (MIC) of Cinnamaldehyde and VRC alone or in combination against 42 clinical Aspergillus fumigatus isolates were determined by both microdillution method and check?board method respectively. The MIC50, MIC90, MICG and MICs distribution were compared between single drug and both in combination. The concentration-accumulative curve was drawn and fractional inhibitory concentration index (FICI) was cal?culated to evaluate the interaction between two test agents. Results The values of MIC50, MIC90 and MICG were significant?ly decreased (P<0.001) when combination of the two drugs than those of their single use, with their MIC distribution concen?tration-accumulative curves shifted to the left. The value of FICI of Cinnamaldehye-VRC combination ranged from 0.187 5 to 1.5. Sixteen strains (38.10%) of them showed the synergistic effect, 19 strains (45.23%) showed additive effect, and 7 strains (16.67%) showed an unrelated effect, and no antagonist effect on tested Aspergillus fumigatus strains in vitro. Conclu?sion Cinnamaldehye in combination with VRC mainly shows a combined synergic and additive inhibitory effect on Asper?gillus fumigatus isolates, and this combination appears to be more active against the test strains, which are less susceptible to voriconazole.

A clinical analysis of twenty-five cases of eosinophilic lung disease / 中华内科杂志

Objective To improve the diagnosis and treatment of eosinophilic lung disease.Methods Patients who were diagnosed with eosinophilic lung disease and hospitalized in the First Affiliated Hospital of Guangxi Medical University Hospital were retrospectively analyzed from January 2004 to August 2012.Data of etiology,clinical manifestation,imaging and pathological features,diagnosis and treatment were recorded.Results A total of 25 patients were diagnosed with eosinophilic lung disease including 9 chronic eosinophilic pneumonia,6 churg-strauss syndrome,and 10 cases of parasitic infection of which two patients were the simple pulmonary eosinophilia (L(o)ffler syndrome).Eosinophil counts in peripheral blood and bronchoalveolar lavage fluid (BALF) were increased.Arterial gas analysis showed varying degree of hypoxemia,which pulmonary function tests showed restrictive,obstructive,mixed ventilatory dysfunction.Chest CT showed bilateral flaky,streak or flake diffuse ground-glass infiltrates and reticular opacities.Results of pulmonary biopsy or skin biopsy identified diffuse eosinophil infiltration.Corticoidsteroid therapy alone or combined with immunosuppressive agents were both effective.Conclusion (1) Liver fluke and other food-borne parasites are the most common causes in eosinophilic lung disease; followed by unexplained chronic acidophilic granulocyte pneumonia; (2) In addition to histopathological evidence,the diagnosis of eosinophilic lung disease was made comprehensively based on clinical features,laboratory test,the BALF analysis,and imaging data.

Caspofungin versus liposomal amphotericin B for treatment of invasive fungal infections or febrile neutropenia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nowadays, there are published trials in regards to the comparison of caspofungin with liposomal amphotericin B (L-AmB). However, these studies have a modest sample size and convey inconclusive results. The aim of this study was to review the efficacy and safety of caspofungin for the treatment of invasive fungal infections (IFIs), compared with L-AmB.</p><p><b>METHODS</b>Electronic databases (up to July 31, 2013) PubMed and Embase databases, the Cochrane Library, and Google Scholar were searched to identify relevant trials of caspofungin and L-AmB. Analyses of efficacy and adverse outcomes were performed by relative risks (RRs) and 95% confidence intervals (CIs). Heterogeneity was assessed by χ(2)-test and the I(2)-statistic.</p><p><b>RESULTS</b>Three trials were included in this meta-analysis with 1249 modified intention-to-treat (MITT) patients. The results showed that caspofungin produced equal efficacy in favorable overall response (RR = 1.02, 95% CI 0.88-1.18; P = 0.81) and mortality rate (RR = 1.53, 95% CI 0.38-6.27, P = 0.55), safer in clinical adverse events (RR = 0.20, 95% CI 0.08-0.54; P = 0.001), laboratory adverse events (RR = 0.69, 95% CI 0. 57-0.84; P = 0.0002), and discontinuation rate (RR = 0.26, 95% CI 0.08-0.83, P = 0.02), compared with L-AmB in the treatment of patients with IFIs.</p><p><b>CONCLUSION</b>Based on the results of this meta-analysis, it would appear that caspofungin was measured to have equal efficacy in clinical outcomes and safer in terms of adverse events.</p>

Prevention of catheter-related Pseudomonas aeruginosa infection by levofloxacin-impregnated catheters in vitro and in vivo / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Implanted medical catheter-related infections are increasing, hence a need for developing catheter polymers bonded to antimicrobials. We evaluated preventive effects of levofloxacin-impregnated catheters in catheter-related Psuedomonas aeruginosa (strain PAO1) infection.</p><p><b>METHODS</b>Drug release from levofloxacin-impregnated catheters was measured in vitro. Levofloxacin-impregnated catheters and polyvinyl chloride (PVC) catheters were immersed in 5 ml 50% Luria Bertani medium containing 10(8) CFU/ml Pseudomonas aeruginosa then incubated for 6, 12, 24 or 48 hours at 37°C when bacteria adhering to the catheters and bacteria in the growth culture medium were determined. Impregnated and PVC catheters were singly implanted subcutaneously in mice, 50 µl (10(7)CFU) of PAO1 was injected into catheters. After the first and fifth days challenge, bacterial counts on implanted catheters and in surrounding tissues were determined microbiologically. Bacterial colonization and biofilm formation on implanted catheters were assessed by scanning electron microscopy.</p><p><b>RESULTS</b>Drug release from levofloxacin-impregnated catheters was rapid. Levofloxacin-impregnated catheters had significantly fewer bacteria compared to PVC in vitro. After first and fifth day of challenge, no or significantly fewer bacteria adhered to impregnated catheters or in surrounding tissues compared to PVC. Scanning electron microscopical images after first day displayed from none to significantly fewer bacteria adhering to impregnated implanted catheters, compared to bacteria and microcolonies adhering to PVC catheters. After the fifth day, no bacteria were found on impregnated catheters, compared to clusters surrounding mucus-like substance and coral-shaped biofilms with polymorphonuclear leukocyte on PVC catheters. After the first day of challenge, secretion occurred in all implanted catheters with surrounding tissues mildly hyperaemic and swollen. After the fifth day, minute secretions inside impregnated catheters and no inflammation in tissues, whereas purulent secretion inside PVC catheters and abscesses in surrounding tissues.</p><p><b>CONCLUSION</b>Levofloxacin-impregnated catheter is a promising new strategy for prevention of catheter-related Psuedomonas aeruginosa infection.</p>

Risk factors and treatment efficiency for lung cancer patients with venous thromboembolism / 实用医学杂志

Objective To investigate the risk factors and treatment efficiency for lung cancer patients with venous thromboembolism (VTE). Methods Total 282 cases of lung cancer patients with VTE were enrolled into two groups , including the VTE group and the non-VTE group , for comparation analysis based on a series of clinical data. Results The occupation rate of adenocarcinoma and Ⅳ period were 65.28% and 87.50% in VTE group, respectively, higher than those of 51.43% and 75.71% in the non-VTE group. The increased rate of blood viscosity and d-dimer respectively were 65.28% and 70.83%, higher than those of 51.43% and 56.67% in the non-VTE group, with significant differences (P < 0.05, respectively). Result of logistic regression analysis showed that tumor stage , d-dimer levels , smoking , age , and blood viscosity levels were highly correlated with venous thrombosis in patients with lung cancer, and the OR value among them was 3.802, 2.339, 5.814, 3.875 and 6.404, respectively, with significant differencees (P < 0.05, respectively). Conclusions Lung adenocarcinoma with stage Ⅳ, smoking , age and increase of blood viscosity and d-dimer were the important risk factors for VTE in patients with lung cancer chemotherapy. Timely assessment of risk factors and early anticoagulation therapy in lung cancer patients with venous thromboembolism associated with VTE can improve the treatment efficacy and reduce the complications.

In vitro inhibitory effects of chlorogenic acid and isochlorogenic acid on Aspergillus fumigatus biofilm / 中国感染与化疗杂志

Objective To investigate the in vitro destructive effect of chlorogenic acid (CRA)/isochlorogenic acid (IRC)on Aspergillus fumigatus biofilm in a model of biofilm formation.Methods The in vitro biofilm model was established using clinical isolates of A.fumigatus.The minimum inhibitory concentrations (MIC)of antimicrobial agents against A.fumigatus were determined by microdilution method.Scanning electron microscope (SEM)and laser confocal scanning microscope (CLSM)were used to characterize the biofilm.Crystal violet staining method was used for biofilm quantitation.Results After reaction with CRA/IRC for 24 h or 48 h,observation of biofilm showed that A.fumigatus extracellular matrix was thinner than the control group.Biofilm quantitation showed that the optical densities were 1.05±0.19,1.14±0.26,0.99±0.14 for CRA group (1 024 mg/L,512 mg/L,256 mg/L);1.39±0.06,1.41±0.06,1.60±0.04 for IRC group (1 000 mg/L,500 mg/L,250 mg/L)at 24 h,and 1.91±0.17 for control group (P<0.05).The quantitation of biofilm showed that the optical densities were 2.25±0.05,2.27±0.05,2.31±0.03 for CRA group,2.26 ± 0.02,2.27±0.02,2.29±0.04 for IRC group at 48 h,and 2.36±0.01 for control group (P<0.05).Conclusions CRA/IRC has some inhibitory effect on the formation of A.fumigatus biofilm.

Diagnostic significance of combination of secretory leukocyte protease inhibitor (SLPI), interferon-γ(IFN-γ) and adenosine deaminase (ADA) for tuberculous pleural effusion / 实用医学杂志

Objective To compare the diagnostic significance of pleural SLPI,IFN-γ and ADA for differenti-ating TPE from pleural effusions with the other etiologies. Methods Pleural effusion samples were obtained from 93 patients who were divided into the following groups: tuberculous pleural effusion,malignant pleural effusion, bacterial pleural effusion and transudative pleural effusion. The pleural effusion and/or serum levels of SLPI , IFN-γand ADA were determined. Results 1.The concentrations of SLPI, IFN-γand ADA in tuberculous pleural effusion was higher than that in malignant group, bacterial group and transudative group. 2. The diagnostic value of SLPI, IFN-γor ADA for the diagnosis of tuberculous PE is high respectively. The combinations of SLPI, IFN-γand/or ADA gained the more valuable diagnostic performance. Conclusion Pleural SLPI, IFN-γand ADA may be helpful for the differential diagnosis of tuberculous pleural effusion and the other pleural effusion. The combinations of SLPI or/and IFN-γor/and ADA further increased diagnostic value.

Study of Cinnamic aldehyde effects on expression of E-cadherin and MMP-9 through SHH signaling pathway in lung adenocarcinoma A549 cells / 中国免疫学杂志

Objective:To investigate Cinnamic aldehyde effects on expression of E-cadherin and MMP-9 and proliferation of lung adenocarcinoma A549 cells,and explore the possible mechanism of Sonic Hedgehog (SHH) signaling transduction.Methods:After co-cultured with Cinnamic aldehyde at the concentration of 0,10,20 and 40 μg/ml for 24 h,48 h and 72 h respectively,A549 cells were tested for their proliferation by MTT assay;E-cadherin and MMP-9 level in the supernatant by ELISA;expression of E-cadherin and MMP-9 mRNA by realtime-PCR with SYBR GreenⅠ;and protein expression by Western blot.Results: ①Cinnamic adehyde with concentration at 10 μg/ml would inhibited proliferation of A 549 cells after 24 hours′treatment;with concentration at 10, 20 and 40μg/ml can affect the proliferation significantly ( P<0.05 );with concentration of 40μg/ml cinnamic adehyde for 72 h,the re-markably inhibition of proliferation in A 549 cells was observed , the highest inhibitory rate was ( 93.782 ±5.036 )%.②Cinnamic aldehyde also increased migration rate of A 549 cells.③Expression of components on Hedgehog signaling pathway in A 549 was higher than that in human HBE cells.Cinnamic aldehyde could increase further upregulate of components expression in Hedgehog signaling pathway of A549 cells.④Secretion level of E-cadherin,mRNA and protein were decreased in A549 cells co-cultured with Cinnamic al-dehyde,while secretion level of MMP-9,mRNA and protein level in A549 cells co-cultured with cinnamic aldehyde were increased.Pre-treatment with 2 nmol/ml cyclopamine,an increasing of secretion level of E-cadherin ,mRNA and protein level in A549 cells was observed,decreasing of secretion level of E-cadhein,mRNA and protein level was also observed in A 549 cells.Conclusion:Cinnamic aldehyde inhibits the proliferation in a time-and dose-dependent manner and effected expression of E-cadherin and MMP-9 through sonic hedgehog signaling pathway in lung adenocarcinoma A 549 cells.

Correlation analysis between the BODE index with fatigue symptoms on COPD patients / 中国实用护理杂志

Objective To explore the correlation between BODE index with fatigue symptoms on COPD patients,and provide new ways for clinical evaluation,prediction,symptom control and establishment of effective management mode.Methods 120 COPD patients of stable stage were selected to be investigated and analyzed by the Fatigue Scale-14(FS-14),six-minute walking distance(6MWD),pulmonary func-tion test and the body mass index (BMI).Dyspnea was measured using the Medical Research Council (MRC) dyspnea scale.Results The fatigue symptoms showed high positive linear correlation with BODE index and mMRC,and significant negative correlation with 6 MWD,FEV1％ and BMI.Conclusions The study shows that fatigue symptoms had a higher prevalence in COPD patients of stable stage.There were a high correlation between fatigue symptoms with the BODE index,mMRC,6MWD,FEV1％ and BMI.The BODE index was a good predictor and evaluation of fatigue symptoms.

Ambroxol reinforce the bactericidal effect of levofloxacin on the Pseudomonas aeruginosa biofilm / 中华急诊医学杂志

Objective To observe the destructive and scavenging effect of ambroxol (AMB) on the biofilm (BF) of Pseudomonas aeruginosa (P.a).To evaluate the synergistically bactericidal effect of AMB along with levofloxacin (LFX) on BF of P.a.Methods The early model (cultured for 3 d) and mature model (cultured for 7 d) of P.a wild strain (PAO1) BF were established,in vitro,respectively.The models were randomly (random number) divided into control group,AMB group and AMB + LFX group.The concentrations of AMB were 256 μg/ml and 512 μg/ml,respectively.When the early BF model and mature BF model were made,different concentrations of AMB were added in AMB group and AMB + LFX (1μg/ml) was added in AMB + LFX group.The number of viable P.a on the BF carrier was counted with the continuous dilution method 24 h after AMB or/and LFX added.Then,the BF morphological changes on the carrier surface were observed by using scanning electron microscopy (SEM).Measured data were analyzed with single factor analysis of variance (One-Way ANOVA).Results Both in the early BF model and in the mature BF model,the SEM examination showed that the BF in AMB group was significantly reduced compared to the control group,and this reduction of BF was in dose-dependent manner.LFX 1 μg/ml could reduce the number of viable bacterial in BF in both early model and mature model (P ＜ 0.05).LFX with addition of different concentrations of AMB showed stronger bactericidal effect than LFX used alone identified by more significant reduction in the number of colonv within the BF (P ＜ 0.05).Furthermore,the LFX combined with 512 μg/ml AMB reduced more significant number of colony apparently than the LFX combined with 256 μg/ml AMB (P ＜ 0.05).Conclusions AMB can destroy the early BF or mature BF partly,and LFX alone can partly reduce the number of viable P.a within BF.When LFX combined with AMB,they exert a synergistically bactericidal effect.

Intervention of azithromycin on Pseudomonas aeruginosa biofilm and virulence factors / 中华微生物学和免疫学杂志

Objective To evaluate the effect of azithromycin on Pseudomonas aeruginosa PAO1 biofilm formation and virulence factors production. Methods Detect the minimum inhibitory concentration of azithromycin against PAO1 by 2-fold dilution method. Crystal violet staining assay was used for initial adhesion assays. The PAO1 biofilm was established in vitro and observed by scanning electron microscope. Viable bacterial counts were determined by serial dilution. LasB elastolytic activity was determined by using Elastin-Congo Red. Protease activity was determined by Azo-casein. Chloroform extraction method was used for pyoverdine assay . The orcinol assay was used to directly assess the amount of rhamnolipids . Results Scanning electron microscope biofilm and viable bacterial counts of PAO1 adhered to the surface of catheter in PAO1 azithromycin group were less than the PAO1 control group after incubated for 3 d and 7 d ( P ＜0.05), and the initial adhesion was weaker ( P ＜ 0. 05 ). The virulence factors production were obviously decreased (P ＜0.01 ). LasB elastolytic activity and pyoverdine were even reduced to the same as with the PA-JP3 group ( P ＞ 0.05 ), but the protease activity and the rhamnolipids concentration were higher than the PA-JP3 group ( P ＜ 0.05 ). Conclusion Azithromycin can inhibit PAO1 bioflim formation in vitro and virulence factors production.

Effect of Meropenem on Biofilms of Pseudomonas aeruginosa lasR/rhlR Mutations in vitro / 中华医院感染学杂志

OBJECTIVE To understand the effects of meropenem on the biofilms of Pseudomonas aeruginosa lasR/rhlR mutations in vitro.METHODS The biofilms of PAEO1 and its lasR,rhlR,and lasR rhlR mutants were incubated in a polyvinyl chloride tube in 0.9% saline for three days,then were immersed in meropenem solution of 26 ?g/ml for 24 hours,and examined by scanning electron microscopy(SEM).RESULTS After three days incubation,PAEO1 biofilms showed a well-developed structure,however,the biofilms of PAEO1 lasR,rhlR,and lasR rhlR mutants were thin and poor developed;after 24 hours in meropenem solution,PAEO1 biofilm become rareness,whereas PAEO1 lasR,rhlR,and lasR rhlR mutants biofilms were almost destructed and only small pieces left.CONCLUSIONS lasR/rhlR Genes play probably an important role in the formation of P.aeruginosa biofilm and in the resistance to meropenem.

Effects of lasR/rhlR Mutation on Biofilm Formation of Pseudomonas aeruginosa in vitro / 中华医院感染学杂志

OBJECTIVE To understand the effects of lasR/rhlR mutations on the biofilm formation of Pseudomonas aeruginosa in vitro.METHODS The biofilms of PAEO1 and lasR,rhlR,and lasR rhlR mutants were incubated in a polyvinyl chloride tube in 0.9% saline and studied by means of scanning electronic microscopy(SEM) after three days′ incubation.RESULTS After 3 day′s incubation,PAEO1 biofilm showed a well-developed structure,whereas thin and poor developed biofilms were seen in lasR/rhlR mutant groups.CONCLUSIONS lasR/rhlR Genes play probably an important role in the formation of P.aeruginosa biofilm in vitro.

Expression of TNF-? in a rat model of chronic Pseudomonas aeruginosa biomembrane associated with lung infection / 中国病理生理杂志

AIM:To investigate the kinetics of tumor necrosis factor alpha(TNF-?)in an animal model of chronic P.aeruginosa biomembrane associated with lung infection.METHODS:Rats were challenged with 0.1 mL of PAO579(1012 CFU/L)in alginate beads or 0.1 mL of planktonic PAO579(1012 CFU/L).After challenging for 3,7 and 14 d,bacteriological and pathological features,and TNF-? expression in lung tissue were observed.RESULTS:(1)CFU/lung in alginate beads group was significantly higher than that in planktonic bacteria group(P

Effects of Quorum-sensing Systems on Chronic Pseudomonas aeruginosa Lung Infection in Rats / 中华医院感染学杂志

OBJECTIVE To understand the importance of quorum sensing(QS)system in Pseudomonas aeruginosa(PAE)biofilm infection in lungs,the pathogenic effects of the wild-type P.aeruginosa PAEO1 were compared with QS double mutants PAEO1 lasR rhlR and PAEO1 lasⅠ rhⅡ in vivo.METHODS Rats were challenged intratracheally with alginate embedded PAE strain PAEO1 and the mutants of QS in the concentration of 1?109 colony-forming units per milliliter(CFU/ml).Two weeks post intratracheal challenge with P.aeruginosa,parameters were evaluated.RESULTS Two weeks after challenge,significantly milder microscopic and macroscopic lung pathology(P