Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents ; pharmacology ; Biofilms ; drug effects ; Candida albicans ; genetics ; physiology ; ultrastructure ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; HSP90 Heat-Shock Proteins ; genetics ; Microscopy, Electron, Scanning ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.

METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.

RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.

CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.

METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.

RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.

CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.

Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology

OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.

METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.

RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.

CONCLUSIONYHN could inhibit C. albicans biofilms in rats.

Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats

To evaluate the regulating effect of Buyang Huanwu decoction and its simple prescription (Naojian tablet) on CDK4/Cyclin D1 expression in hippocampus tissues of rats with cerebral ischemia, SD rats were divided into the sham-operation group, the model group, the Buyang Huanwu decoction group (ig, 3.15 g · kg⁻¹) and the simple prescription group (ig, 2.41 g · kg⁻¹). Each group was further divided into five subgroups based on time points after the administration, i. e. 1 d, 3 d, 7 d, 14 d and 28 d, respectively. CDK4/Cyclin D1 expressions of the group at different time points were examined by using immunohistochemistry and real-time qPCR. According to the results, the cerebral ischemia model group showed higher CDK4/Cyclin D1 expression than the sham-operation groups (P < 0.05), suggesting that the cell cycle signal pathway would be activated by the cerebral ischemic injury. Both Buyang Huanwu decoction and simple prescription groups showed significantly lower cyclin expression than the model group at 3 d, 7 d, 14 d, 28 d (P < 0.05), indicating both Buyang Huanwu decoction and its simple prescription could play the neuroprotective effect through the cell cycle signal pathway.
Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Cycle ; drug effects ; Cyclin D1 ; genetics ; metabolism ; Cyclin-Dependent Kinase 4 ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on hyphae development of Candida albicans.

METHODInverted microscope, fluorescence microscope, SEM were applied to inspect the Morphological change of C. albicans treated by EAHD at different concentrations. Solid agar plate was utilized to evaluate the colony morphology. Quantitative Real-ime PCR(qRT-PCR) was adopted to observe the expression of hyphae-specific genes such as HWP1, ALS3, UME6, CSH1, SUN41, CaPDE2.

RESULTEAHD with concentration of 312 and 1 250 mg . L-1 could inhibit formation of hyphae and colony morphology. The expression of HWP1, ALS3, UME6, CSH1 were downregulated 4. 13, 3. 64, 2. 46, 2. 75 folds ,while the expression of SUN41 were upregulated 7. 26 folds, CaPDE2 keep unchanged.

CONCLUSIONEAHD could inhibit formation of hyphae and colony morphologies of C. albicans through downregulating HWP1, ALS3, UME6 and CSH1.

Acetates ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; genetics ; growth & development ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; Hyphae ; Medicine, Chinese Traditional ; Microscopy, Fluorescence ; Reverse Transcriptase Polymerase Chain Reaction

Objective To observe the changes of endothelin-1 (ET-1) and prostaglandin E2 (PGE2) in exhaled breath condensate (EBC) and serum of patients with acute respiratory distress syndrome (ARDS) after treated by Qingfeitang and investigate its clinical value. Methods 52 ARDS patients receiving mechanical ventilation at intensive care unit (ICU) were divided into the Qingfeitang treatment group and the control group, with 26 cases in each group. The EBC were collected by Ecoscreen condenser within 24 h after diagnosis of ARDS and on the 5th day of medication, and the venous blood were collected at the same time. The levels of ET-1 and PGE2 in the EBC and serum of different period were measured by EIA. Results (1) After treatment, The levels of ET-1 in EBC and serum of the Qingfeitang treatment group were significantly lower than those of the control group. (2) After treatment, the levels of PGE2 in serum of the Qingfeitang treatment group were significantly lower than that of the control group. (3) The oxygenation index difference before and after treatment of Qingfeitang in the treatment group was higher than in the control group. (4) The duration of mechanical ventilation of the Qingfeitang treatment group was significantly less than that of the control group. Conclusions Qingfeitang could be an effective method for alleviating acute respiratory distress syndrome.

Objective To know the prevalence of chronic obstructive pulmonary disease(COPD) in patients with obstructive sleep apnea-hypopnea syndrome(OSAHS) in Kunming Yunnan Province, and the clinical symptom of OS.Methods Retrospective study of 4 636 cases of patients with snoring, excluding COPD in addition to chronic respiratory disease and assess the patient`s condition.The test included AHI, BMI, Epworth sleepiness scale, lung function.The index of OS was confirmed by AHI>5 times/h and FEV1/FVC<70%.Results During the period of 2006 to 2012, he prevalence of COPD in patients with OSAHS was 10.1% [95% confidence interval (CI) 9.1%-11.1%] in Kunming Yunnan Province.And male`s OS prevalence rate is higher than the female(male 10.2%;female 9.7%).The OS patients` average age was 56.9±14.1, the mean AHI was (47.46±26.79) times/h, and the average FEV1/FVC was 60.09%±23.57%.Polysomnographyresults show that patients suffered obvious hypoxiaat night in both OSAHS group and OS group, but it was more significant in OS group.Pulmonary function test showed that OS patients have more serious chronic obstructive pulmonary disease.Conclusions The prevalence of COPD in OSAHS patients was high in Kunming, Yunnan Province, and the prevalence rate in old group reached more than 24%.In addition, the sympotms of patients with OS were more severe than those with only OSAHS or COPD in lung function and hyoxemia.

Objective To investigate the changes of gastric emptying in awake rats after acute myocardial infarction(AMI). Methods The male Wistar rats were submitted to thoracotomy followed by or not by occlusion of the left coronary artery. After 24 h, AMI(n=12)and sham groups (n=9)were fed in gavage(1 ml)with ~(99m)-Tc-DTPA labeled test meal. Half gastric emptying time (GET1/2) and 50min residual rate were measured by single photon emission computed tomography. The rats of AMI group were sacrificed after 50 min. Plasma level of motilin was measured by radioimmunoassay in AMI and sham group respectively(n=9). Results GET1/2 was significantly higher in AMI group[(23.1?4.7)min] than in sham group[(16.0?4.0)min](P

There are reports about the chemical compounds of Ciwujia herbs, but with no study report about the chemical material basis of Ciwujia injection (CWJI). In this study, LC-MS(n) and LC-Q-TOF-MS techniques were adopted for a qualitative analysis on phenylpropanoids in CWJI. The Ultmate XB-C18 column (4.6 mm x 250 mm, 5 microm) was adopted and eluted with the mobile phase of 0.5% formic acid-water and acetonitrile, with the flow rate at 0.8 mL x min(-1) and the column temperature at 20 degrees C. Based on the data of high-resolution and multi-stage MS, control products and literatures, altogether 54 phenylpropanoids were identified in Ciwujia Injection, including 34 phenylpropanoids, 16 ligans and 4 coumarins. Among them, 28 were reported for the first time in Ciwujia, and 14 compound structures were identified in comparison with the control products. The method established in this study could be used to simply and rapidly identify phenylpropanoids in CWJI. The findings provide scientific data for defining the chemical material basis of CWJI.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Eleutherococcus ; chemistry ; Mass Spectrometry ; methods ; Molecular Structure