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 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 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

Objective To investigate and compare the effects of jejunal interposition and Roux-en-Y surgery after total gastrectomy. Methods The clinical data of 41 patients with gastric cancer who underwent total gastrectomy in our hospital from June 2013 to June 2016 were retrospectively analyzed.According to the different ways of reconstruction of digestive tract,the patients were divided into observation group and control group.Patients of the observation group were with jejunal interposition surgery,while patients of the control group were trea-ted with Roux-en-Y surgery,and the effects of the two groups were compared and analyzed.Results There was no significant difference be-tween the two groups in the time of reconstruction,length of hospital stay and the incidence of complications(P>0.05).One year after sur-gery,incidence of complications,albumin,total health score,fatigue score,and disgust score of the observation group were significantly better than those of the control group with statistically significant difference(P<0.05).There was no significant difference between the two groups in survival rate,hemoglobin,total protein,weight changes(P>0.05).Conclusion For patients with total gastrectomy,jejunal interposition and Roux-en-Y surgery had similar effect.But in terms of the long-term effect,jejunal interposition can reduce complications and improve lev-el of nutrition and quality of life.

OBJECTIVETo investigate the effect of andrographolide (AG) on quroum sensing (QS) and relevant virulence genes of Candida albicans.

METHODGas-chromatography-mass spectrometry (GC-MS) was applied to detect the changes in the content of farnesol and tyrosol in C. albicans intervened by AG. The real-time quantitative PCR (qRT-PCR) was adopted to inspect the expressions of relevant virulence genes such as CHK1, PBS2 and HOG1 regulated by QS.

RESULTAt 2 h after the growth of C. albican, the farnesol and tyrosol secretions reduced, without notable change after intervention with AG. The secretions were highest at 12 h and decreased at 24 h. After the intervention with different concentrations of AG, the farnesol content reduces, whereas tyrosol increased, indicating a dose-dependence, particularly with 1 000 mg x L(-1) AG. qRT-PCR revealed that 1 000 mg x L(-1) AG could down-regulate CHK1 by 2.375, 3.330 and 4.043 times and PBS2 by 2.010, 4.210 and 4.760 times, with no significant change in HOG1.

CONCLUSIONAG could inhibit the farnesol secretion, promote the tyrosol secretion and down-regulate QS-related virulence genes CHK1 and PBS2 expressions.

Candida albicans ; drug effects ; genetics ; physiology ; Diterpenes ; pharmacology ; Farnesol ; analysis ; metabolism ; Gas Chromatography-Mass Spectrometry ; Genes, Fungal ; Phenylethyl Alcohol ; analogs & derivatives ; analysis ; metabolism ; Quorum Sensing ; drug effects ; Real-Time Polymerase Chain Reaction ; Virulence ; genetics

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

OBJECTIVETo clarify the effect of histamine H4 receptor antagonist, JNJ 7777120, and histamine H1 receptor antagonist, Loratadine, on allergic rhinitis (AR) in rats and to study the role of histamine H4 receptor antagonist and histamine H1 receptor antagonist in the pathogenesis of allergic rhinitis and therapeutic value of their antagonist.

METHODSAR animal model were induced by ovalbumin (OVA) in the Wistar rats, which treated with histamine H4 receptor antagonist and (or) histamine H1 receptor antagonist. The allergic symptoms (sneezing and nasal rubbing), serum total IgE and the levels of cytokines in serum or nasal lavage fluid were measured, the diversity between two groups were observed. Statistical analysis was performed using a SPSS 13.0 software.

RESULTSCompared with AR group with no treatment, the inhibition of nasal symptoms (P < 0.01), a significant decrease in the levels of IgE, IL-4 in serum and Eotaxin in nasal lavage fluid (P < 0.01), a significant increase in the levels of IFN-γ in serum (P < 0.01) after treatment was found. Compared with group treated with Loratadine, inhibition of nasal symptoms (q value were 3.72, 4.16, P < 0.01), a significant increase in the levels of IgE and IL-4 in serum (q value were 8.01, 4.96, P < 0.05), a significant decrease in the levels of IFN-γ in serum (q = 3.18, P < 0.05) in group treated with JNJ 7777120 also, but no significant differences in the levels of Eotaxin in nasal lavage fluid (P > 0.05). Administration of JNJ 7777120 and Loratadine jointly, neither additive effect nor synergistic action were found (P > 0.05).

CONCLUSIONSHistamine H4 receptor is closely related with allergic rhinitis and is important in the pathogenesis of allergic rhinitis, the same as histamine H1 receptor. Histamine H4 receptor antagonist, JNJ 7777120, could relieve symptoms and inflammatory conditions in allergic rhinitis, the effect was weak compared with Loratadine. Neither additive effect nor synergistic action were found between them.

Animals ; Histamine Antagonists ; pharmacology ; therapeutic use ; Indoles ; pharmacology ; therapeutic use ; Loratadine ; pharmacology ; therapeutic use ; Male ; Piperazines ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar ; Receptors, Histamine ; metabolism ; Rhinitis, Allergic, Perennial ; drug therapy ; metabolism

OBJECTIVETo investigate the effects of different-intensity sinusoidal electromagnetic fields (SEMFs) on bone mineral density (BMD) and histomorphometry in SD rats.

METHODSThirty female SD rats were randomly divided into three groups: group A (a control group), group B (0.1 mT group) and group C (0.6 mT group). The rats in group B and C were exposed to 50 Hz SEMFs 3 hours each day. However,the magnetic intensity was different between group B and group C:0.1 mT for group B and 0.6 mT for group C. After 8 weeks, all the animals were killed. Changes of BMD and histomorphometric properties were observed.

RESULTSCompared with group A, the BMD of whole body, femur and vertebrae of rats in group B increased significantly; the area percentage, number and width of bone trabeculae in vertebrae and femur of rats in group B were larger than those of group A; but the resolution of bone trabeculae of rats in group B was lower than that of group A. The trabecular number in group C rats were significantly decreased, compared with that in group A rats. The outcome of double fluorescence labeling in group B was found to be significantly different with that in group A. But the difference between rats in group A and C was not significant.

CONCLUSIONThis study demonstrates that 50 Hz 0.1 mT SEMFs can increase BMD, improve bone tissue microstructure and, promote bone formation.

Animals ; Bone Density ; radiation effects ; Electromagnetic Fields ; Female ; Lumbar Vertebrae ; pathology ; radiation effects ; Osteogenesis ; radiation effects ; Rats ; Rats, Sprague-Dawley ; Tibia ; pathology ; radiation effects

OBJECTIVETo develop attenuated Salmonella which harboring enterovirus 71 (EV71) VP1 gene.

METHODSThe plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis. The recombinant plasmid was then transformed into attenuated Salmonella SL7207.

RESULTSEV71 VP1 gene sequence was inserted into a eukaryotic expression plasmid VR1012. VP1 protein was detected by Western Blot analysis in the culture supernatant. And the attenuated Salmonella harbored the plasmid stable.

CONCLUSIONThe plasmid was constructed successfully and it can express effectively in vitro. The bacteria which harboring the plasmid were constructed successfully. This has provided a basis for further research of an oral EV71 vaccine.

Capsid Proteins ; genetics ; metabolism ; Enterovirus A, Human ; genetics ; Gene Expression ; Genetic Engineering ; Genetic Vectors ; genetics ; metabolism ; Salmonella ; genetics ; metabolism

OBJECTIVETo study the feasibility of inducing the differentiation of rat adipose-derived stem cells (ADSCs) into smooth-muscle-like cells in monolayer culture in vitro.

METHODSADSCs were obtained from the adipose of the inguinal region of the SD rat. A growth curve of the ADSCs was drawn. The fourth passage ADSCs were induced to differentiate into adipocytes with adipogenic inducing fluid and determined by Oil Red O staining, into osteoblasts with osteogenic inducing fluid and determined by Von Kossa staining, as well as into smooth-muscle-like cells with inducing fluid containing beta-mercaptoethanol, and the expression of alpha-smooth muscle actin (alpha-SMA) was detected by the immunohistochemical method.

RESULTSThe ADSCs presented spindle and polygon shapes and proliferated rapidly in vitro. The growth curve showed the ADSCs in the logarithmic growth phase two days after subculture. The fourth passage ADSCs exhibited red stained lipid droplets characteristic of adipocytes in the cytoplasm on Oil Red O staining after induction with adipogenic inducing fluid, and calcium nodes on Von Kossa staining after induction with osteogenic inducing fluid. The immunohistochemical results of the ADSCs induced into smooth-muscle-like cells showed that the positive rate of alpha-SMA in the beta-mercaptoethanol induced cells was (29.80 +/- 6.89)%, significantly higher than in the uninduced ones ([2.89 +/- 1.24]%, P < 0.01).

CONCLUSIONADSCs displayed obvious characteristics of smooth muscle cells after induction, and could be a new source of cells in the tissue engineering studies of smooth muscle related diseases.

Adipocytes ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; Male ; Myocytes, Smooth Muscle ; cytology ; Rats ; Rats, Sprague-Dawley