Objective: To investigate the effects of butanol extract from Paris polyphylla var. yunnanensis (BEPP) on the biofilm formation of Candida albicans in vitro. Methods: Microdilution method was used to determine the minimum inhibitory concentration (MIC). XTT assay was used to determine the SMIC80. Flat band method was used to protract the time-kill curve. Scanning electron microscopy (SEM) was used to observe the morphological changes of the biofilm. Confocal laser scanning microscopy (CLSM) was used to detect the thickness of the biofilm. The quantification real-time PCR (qRT-PCR) was used to detect the expression changes of genes (EAP1, MP65, and SUN41) of the biofilm cells. Results: The MIC of BEPP against C. albicans strains was determined as 32-128 μg/mL. The SMIC80 of BEPP against the biofilm of C. albicans strains was determined as 128-512 μg/mL. Time-sterilization curve results indicated that BEPP had a promise bactericidal effect. SEM results showed that the formation of C. albicans biofilm was inhibited by BEPP, and the morphology of biofilm was also affected by BEPP. The thickness of C. albicans biofilm was reduced by BEPP according to CLSM results. Furthermore, qRT-PCR results indicated that expressions of EAP1, MP65, and SUN41 were significantly down-regulated by BEPP. Conclusion: Our results demonstrate that BEPP inhibits the biofilm formation of C. albicans in vitro.

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 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

OBJECTIVETo investigate the effects of butyl alcohol extract of baitouweng decoction (BAEB) on the fungal cell surface hydrophobicity (CSH), filamentation and biofilm formation of Candida tropicalis.

METHODGradual dilution method was used to determine the MIC. XTT assay was applied to determine the SMIC80. Time-Kill assay was employed to draw the Time-Kill curve. The water-hydrocarbon two-phase assay was used to measure the cell surface hydrophobicity. Scanning electron microscopy (SEM) was applied to observe the morphological changes of the biofilm. Confocal laser scanning microscopy (CLSM) was applied to determine the thickness of the biofilm. The quantification real-time PCR (qRT-PCR) was used to detect expression changes of releated genes (UME6, ALST3 and NRG1). result: The MICs of BAEB against C. tropicalis strains are determined as 64-128 mg x L(-1). The SMIC80 s of BAEB against the biofilm of Candida tropicalis strains are determined as 256-512 mg x L(-1). Time-Kill curve results indicate that BAEB has a promise fungicidal effect at 256 and 512 mg x L(-1). SEM results shows that 512 mg x L(-1) BAEB can inhibit the formation of C. tropicalis biofilm on Silicone catheter, and the morphology of biofilm is also affected by BAEB. The thickness of C. tropicalis biofilm is reduced by BAEB according to CLSM results. Furthermore, qRT-PCR results indicate that expression of UME6 and ALST3 are significantly down-regulated by BAEB 256,512 mg x L(-1), and NRG1 is not affected by BAEB.

CONCLUSIONBAEB inhibits effectively the CSH, filamentation and biofilm formation of VVC strains of C. tropicalis.

Antifungal Agents ; chemistry ; pharmacology ; Biofilms ; drug effects ; Candida tropicalis ; drug effects ; genetics ; physiology ; Candidiasis ; microbiology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Fungal ; drug effects ; Humans ; Virulence Factors ; genetics ; metabolism

OBJECTIVETo explore a new method to reconstruct the defect of the distal foot.

METHODSA distally based dorsum pedis island flap pedicled with the first dorsal metatarsal artery was designed and transferred to the defect of the distal foot.

RESULTSFive patients were treated with this flap, which ranged from 2 cm x 4 cm to 6 cm x 7 cm in size. Four flaps survived completely, one flap had marginal necrosis and healed after conservative therapy.

CONCLUSIONSThe reverse first dorsal metatarsal artery flap, with good blood circulation and easy manipulation, is a good option for the defects of the distal foot.

Adolescent ; Adult ; Child ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Metatarsus ; blood supply ; Middle Aged ; Skin Transplantation ; methods ; Surgical Flaps ; blood supply ; Young Adult

OBJECTIVETo improve the survival of mesenchymal stem cells (MSCs) for prefabrication of an osteo-musculo-cutaneous flap.

METHODSIn a mouse model, the compound of MSCs and p(3HB-co-3HH) were embeded in the latissimus dorsi muscle as an experimental group and the muscle pocket of the buttock as the control. The examinations of the HE staining, hybridization in situ of osteonectin mRNA and Von kossa staining were used to evaluate the results.

RESULTSThe expression of osteonectin mRNA and the Von kossa staining showed that the latissimus dorsi muscle group was superior to the control in 2 weeks and 4 weeks after the surgery in vivo.

CONCLUSIONThe results indicate that the above-mentioned technique may be a good alternative for the prefabrication of the osteo-musculo-cutaneous flap.

Animals ; Fascia ; transplantation ; Histocytochemistry ; methods ; In Situ Hybridization ; Male ; Mice ; Mice, Nude ; Models, Animal ; Osteonectin ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Staining and Labeling ; Stem Cell Transplantation ; methods ; Surgical Flaps ; blood supply ; Tissue Engineering ; methods

OBJECTIVETo observe the effect of 11,12-epoxyeicosatrienoic acid (11,12-EET) on nitric oxide synthase (NOS) in myocardial ischemia/reperfusion injury and explore the protective role of NOS in myocardium.

METHODSRat myocardial ischemia/reperfusion model was produced by ischemia for 60 minutes and reperfusion for 30 minutes. Rats were divided into 5 groups: 11,12-EET ischemia/reperfusion groups (including EET1, EET2, and EET3 groups), EET control group, ischemia/reperfusion group, sham operation group, and control group. Changes of the maximal rates of rise and decrease of left ventricular pressure (+/-dp/dtmax) were observed. Activities of inducible nitric oxide synthase (iNOS) and constrictive nitric oxide synthase (cNOS) in myocardium were measured with chemocolorimetry.

RESULTSDuring both ischemia period (60 min) and reperfusion period (30 min), +/-dp/dtmax was significantly lower in ischemia/reperfusion group than in sham operation group (P < 0.01), and was significantly higher in EET1, EET2 and EET3 groups than in ischemia/reperfusion group (P < 0.01). cNOS level was significantly lower in ischemia/reperfusion group than in sham operation group, was significantly higher in EET1, EET2 and EET3 groups than in sham operation group (P < 0.01), and was significantly higher in EET2 group than in EET group (P < 0.01). iNOS level was significantly higher in sham operation group than in EET control group (P < 0.05), was significantly higher in ischemia/ reperfusion group than in sham operation group (P < 0.01), and was significantly lower in EET1, EET2 and EET3 groups than in ischemia/reperfusion group (P < 0.01).

CONCLUSIONExogenous 11,12-EET can improve ischemia/reperfusion injury, which may be related with the changes of NOS isozymes.

8,11,14-Eicosatrienoic Acid ; analogs & derivatives ; pharmacology ; Animals ; Male ; Myocardial Reperfusion Injury ; enzymology ; Myocardium ; enzymology ; Nitric Oxide Synthase ; drug effects ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo observe the inhibitory effect of different extract fractions from Longdan Xiegan decoction on biofilms of Candida albicans, and discuss its possible mechanism.

METHODThe micro-dilution method and the XTT reduction assay were adopted to explore the antifungal activity of different extract fractions from Longdan Xiegan decoction and detect the inhibitory effect of different extracts on biofilms of C. albicans. The expression quantity of the adhesion related gene ALS1 and hypha formation SUN41 were detected by qRT-PCR.

RESULTThe MICs of extracts from Longdan Xiegan decoction, including petroleum ether, water, butanol, methanol and ethyl acetate, against C. albicans were > 1 000, > 1 000, > 1 000, 125, 125 mg x L(-1). The SMIC50 against biofilms of C. albicanswere > 1 000, > 1000, > 1 000, 500, 500 mg x L(-1). The SMIC50 were > 1 000, > 1 000, > 1 000, > 1 000 and 1 000 mg x L(-1). 1 000 mg x L(-1) ethyl acetate extracts could considerably inhibit the expression of the adhesion related gene ALS1 and hypha formation SUN41.

CONCLUSIONThe ethyl acetate extract showed the greatest activity against Candida albicans biofilms.

Antifungal Agents ; pharmacology ; Biofilms ; drug effects ; growth & development ; Candida albicans ; drug effects ; growth & development ; Candidiasis ; drug therapy ; microbiology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Hyphae ; drug effects ; growth & development ; Microbial Sensitivity Tests

BACKGROUNDAtrioventricular nodal reentrant tachycardia (AVNRT) is one of the most common paroxysmal supraventricular tachyarrhythmias. The aim of the study was to prospectively compare the characteristics of radiofrequency catheter ablation of AVNRT guided by a magnetic navigation system with the conventional procedure.

METHODSPatients with AVNRT diagnosed by electrophysiological tests were randomized into two groups. In the conventional technique group (CMT), a common 4-mm-tip quadrapolar temperature-controlled ablation catheter was used. In the magnetic navigation system guidance group (MNS), a magnetic 4-mm-tip quadrapolar temperature-controlled ablation catheter was used. The following parameters were collected and compared between the two groups: ablation procedure time, patient fluoroscopy time, operator fluoroscopy time, energy delivery numbers, maximal energy per deployment, success rate, complication rate and operative cost.

RESULTSForty patients were enrolled and randomized into CMT and MNS groups. The age, gender, tachycardia history and basic cardiovascular diseases of the two groups were comparable (P > 0.05). All procedures were conducted successfully without complications. No tachycardia recurred during the follow-up period of (9.3 ± 2.6) months. In the MNS group, the patient and operator fluoroscopy times ((11.5 ± 4.3) min, (4.2 ± 1.5) min), energy delivery numbers (3.2 ± 0.9), and maximal energy per deployment ((16.9 ± 3.4) W) were shorter or lower than those of the CMT group ((14.3 ± 6.2) min, (13.6 ± 3.5) min, 6.3 ± 2.1, (23.7 ± 1.3) W, respectively) (P < 0.05). But the operative cost for the MNS group was higher than that of the CMT group (P < 0.01).

CONCLUSIONMagnetic navigation system guided radiofrequency catheter ablation of AVNRT has the advantages of shorter fluoroscopy time and lower energy delivery numbers and maximal energy per deployment compared to the present conventional ablation technique.

Catheter Ablation ; methods ; Humans ; Tachycardia, Atrioventricular Nodal Reentry ; surgery ; Treatment Outcome