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 identify the interaction between nuclear localization signal-retinoic acid receptor α (NLS-RARα) and Ubiquilin 1(UBQLN1). Methods The recombination expression plasmids pGBKT7-NLS-RAR and pACT2-UBQLN1, which expressed bait-protein NLS-RARα and target protein UBQLN1 respectively, were cotransformated into yeast AH109. The interaction of the expression plasmids in the living cells was investigated by yeast two-hybrid assay. HA-tagged fusion protein (pCMV-HA-NLS-RARα) expression vector and Myc-tagged fusion protein (pCMV-Myc-JTV1) expression vector were constructed, identified, and cotransfected respectively into human embryo kidney 293 cells(HEK293). The interaction was detected by co-immunoprecipitation.Results Blue clones were found on yeast AH109 plate cotransformated with pGBKT7-NLS-RARα and pACT2-UBQLN1. Double restriction enzyme digestion showed that pCMV-HA-NLS-RARα and pCMV-Myc-JTV1 were successfully constructed. Then HA-NLS-RAR protein was immunoprecipitated by anti-HA polyclonal antibody and the expression of Myc-UBQLN1 was tested by Western blot with anti-Myc monoclonal antibody from immunoprecipitated complex. Conclusion The interaction between NLS-RARα and UBQLN1 can be verified by yeast two-hybrid assay and co-immunoprecipitation.

Poly ( duenedumethylammonuum chlorude ) ( PDDA ) functuonaluzed sulver nanopartucles ( AgNPs ) prepared wuth PDDA as the protectuve and reductuve agents was combuned wuth graphene oxude ( GO ) to prepare PDDA functuonaluzed cubuc sulver nanopartucles ( C-AgNPs)/GO composute, whuch was then modufued on a glassy carbon electrode ( GCE) to form C-AgNPs-PDDA/GO/GCE. The surface morphologues of dufferent modufued electrodes were characteruzed by scannung electron mucroscope ( SEM ) , and theur correspondung cycluc voltammetruc ( CV) behavuors were unvestugated, unducatung that the composute of C-AgNPs-PDDA/GO exhubuted excellent electrocatalytuc oxudatuon actuvuty to DA and NO-2 . By usung dufferentual pulse voltammetry, the responses of C-AgNPs-PDDA/GO/GCE were lunear un the ranges of 0 . 030-0 . 300 μmol/L and 0 . 300-300 μmol/L wuth detectuon lumut of 9. 8 nmol/L (S/N=3) for DA, and 30. 0-2300 μmol/L wuth detectuon lumut of 12. 6 μmol/L (S/N=3) for NO-2, respectuvely. The modufued electrode dusplayed good selectuvuty, reproducubuluty and stabuluty, and could be used for the sumultaneous determunatuon of DA and NO-2 un human serum samples wuth recoverues of 97. 4%-104. 2% and 98. 0%-102. 8%, respectuvely. Compared wuth spectrophotometruc method, the determunatuon results were satusfactory, showung that the modufued electrode possessed a potentual applucatuon value.

Objective To investigate the incidence and risk factors of acute kidney injury (AKI)in hepatitis B virus (HBV)related acute-on-chronic liver failure (ACLF)patients,and to explore the impact of AKI on the prognosis of ACLF.Methods The medical records of 227 patients who were diagnosed with HBV-related ACLF at the Department of Infectious Diseases in the First Affiliated Hospital of Nanchang University from January 2015 to August 2016 were retrospectively reviewed.Patients were divided into AKI group and non-AKI group based on the AKI criteria published by International Club of Ascites in 2015 .Demographic and clinical data were compared between groups.The AKI incidence and its impact on patients’prognosis were analyzed.The comparison of continuous variables was done by t test or rank-sum test.The comparison of categorical variables was done byχ2 test or Fisher exact test.AKI risk factors were analyzed by using logistic regression.Results There were 66 (29.1 %)cases were diagnosed with AKI among 227 ACLF patients,among which,45 patients (68.2%)were stage Ⅰ,14 (21 .2%) were stage Ⅱ and 7 (10.6%)were stage Ⅲ.Age,cirrhosis,concentrations of total bilirubin and albumin,international normalized ratio (INR),percentage of neutrophils,MELD scores and spontaneous peritonitis rate (SBP)were all statistically different between AKI group and non-AKI group (all P <0.05).The binary logistic regression analysis revealed that only INR (OR=3.132,P =0.001 )and SBP (OR=4.204,P =0.001 )were the independent risk factors of AKI.The optimal cut-off value for INR was 2.025 with AUROC of 0.609 (P =0.01),sensitivity of 59.1 % and specificity of 62.1 %.The 30-day mortality of AKI group was significantly higher than non-AKI group (χ2= 18.324,P < 0.01). Conclusions AKI is relatively common in patients with ACLF.The risk factors of AKI are INR and SBP. AKI has significant impact on the short-term survival rate of ACLF.Therefore,physicians should pay attention to patients with INR of ACLF at admissions and SBP during the management so as to prevent the occurrence of AKI and to reduce the fatality of ACLF.

Objective To evaluate the value of electrospun polymethyl methacrylate (PMMA) nanofibers with different topological structures as scaffolds for growth of Schwann cells (SCs).Methods Electrospun PMMA nanofibers with random or aligned topological structures were fabricated and measured with biocompatibility.Lentivirus-transfected green fluorescent protein was used as the reporting gene to monitor form and growth manner of SCs on different substrates and dependency of cell body and process with fiber structure,with PMMA thin films served as the control.Results Electrospun PMMA nanofibers revealed good biocompatibility and could exert contact guidance to the growth of SCs.Topological structures of the electrospun nanofibers influenced cell morphology.SCs were aligned with the orientation of substrate fibers and form longer cell process when growing on aligned nanofibers (P ＜0.01).Primary SCs preferred to follow the cue of aligned nanofibers compared to random fibers.Conclusion Aligned electrospun PMMA nanofibers have the potentiality as transplantable scaffolds for loading SCs after neural injury.

OBJECTIVETo establish a quantitative method to detect circulating tumor cells (CTC) in patients with small cell lung cancer, and analyze its sensitivity and stability.

METHODSA specific primer and probe for prepro-gastrin-releasing peptide (preproGRP) was designed and a quantitative RT-PCR method was established to detect preproGRP mRNA. Cell incorporation method was used to evaluate the sensitivity. Magnetic cell sorting (MACS) was used to isolate and purify CTC from peripheral blood, and the MACS in combination with morphological diagnosis were used for cell counting.

RESULTSThe isolation rate of CTC by MACS was 30% and the lower detection limit was 5 cells per ml blood. The sensitivity of quantitative RT-PCR in detection of preproGRP mRNA in CTC was 0.64 cells per reaction, and the lower detection limit was 50 cells per ml blood, which was lower than that of MACS. However, the cell numbers calculated by Ct value was in greater accordance (about 80%) with actual cell numbers than that obtained by MACS.

CONCLUSIONSPreproGRP quantitative RT-PCR and MACS have both advantages and disadvantages in detecting CTC of SCLC patients. MACS has a higher sensitivity, and is more favorable when CTC count is below 50 per ml blood. Meanwhile, preproGRP mRNA quantitative RT-PCR is more reliable in calculating actual cell numbers.

Humans ; Immunomagnetic Separation ; Lung Neoplasms ; blood ; metabolism ; pathology ; Neoplastic Cells, Circulating ; Peptides ; genetics ; metabolism ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Small Cell Lung Carcinoma ; blood ; metabolism ; pathology

Infective endocarditis (IE) remains a serious disease. Aorta-to-right atrium fistula is a rare but very serious complication of IE and predicts a higher mortality. This report describes a 50-year-old man with endocarditis, vegetation, perforation of noncoronary sinus, and formation of two aorta-to-right atrium fistulas with native valves detected by transthoracic echocardiography. This disease is lethal despite developments in cardiac imaging and antibacterial therapy. Early diagnosis, aggressive antibacterial therapy, and surgical treatment may improve the prognosis.
Aorta ; diagnostic imaging ; Arterio-Arterial Fistula ; complications ; diagnostic imaging ; Echocardiography, Doppler ; Endocarditis ; complications ; diagnostic imaging ; Heart Atria ; diagnostic imaging ; Humans ; Male ; Middle Aged

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 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

Objective To investigate the current status of secondary prevention of cerebral infarction in Longquanyi District of Chengdu. Methods Prospective Investigation on the secondary prevention was conducted in 94 pa-tients with cerebral infarction at 3, 12 and 24 months following stroke. Results The percentages of patients receiving anti-platelet therapy, antihypertensive treatment, antidiabetic drug treatment and statin therapy were 96.8%, 92.1%, 91.7%and 63.8%at hospital discharge, 79.8%, 88.9%, 91.7%and 55.3%at 3 months, 76.1%, 75.0%and 41.4%, 63.8%at 12 months and 33.0%, 71.4%;58.3%and 22.3%at 24 months, respectively. None of stroke patients with atrial fibrillation re-ceived anticoagulation therapy. Conclusion There is a large gap between the current practice of secondary prevention and guideline in patients with cerebral infarction in Longquanyi District of Chengdu.