Objective:To study the role of RapIDYeast Plus (RYP) system in identifying common clinical yeasts.Methods: The target strains were cultured and passaged twice on Sabouraud dextrose agar,and were fed to RYP system after 24-48 h incubation at 30℃.Results: One hundred and thirty-nine of the 150 target strains were identified to the level of species correctly,and 8 undetermined strains were confirmed by additional tests.It was found that 3 strains had been incorrectly identified by RYP system.The accuracy of RYP system was calculated as 92% without additional tests and 98% with additional tests.Conclusion: RYP system is suitable for routine tests in clinical microbiological laboratory;it can accurately identify more than 40 kinds of yeasts and yeast-like bacteria in clinical practice.

Diagnostic Errors ; Encephalocele ; diagnosis ; Female ; Humans ; Meningocele ; diagnosis ; Middle Aged ; Mucocele ; diagnosis ; Sphenoid Sinus ; pathology

Objective To investigate the effects of heat-inactivated Cryptococcus neoformans(H-CN)on an experimental murine model of meningoencephalitis and on IL-1?,IFN-?and TNF-?gene ex-pression on the brain and spleen.Methods An experimental murine model of intracerebral infection with C.neoformans was established.Mice were divided into H-CN-treated group and control group.The brain and spleen of two groups were collected to obtain total RNA,and IL-1?,IFN-?and TNF-?were detected by RT-PCR method.After intracerebral challenging with lethal doses of C.neoformans,the survival time and colony forming units(cfu)of C.neoformans in the brain of two group were observed.Results The survival time was prolonged,and cfu of C.neoformans were decreased in the brain of H-CN-treated group in com-parison with those of control group.Expression of IL-1?was positive,and IFN-?and TNF-?negative in the brain tissue of H-CN-treated mice;while expression of IL-1?,IFN-?and TNF-?was all negative in the control mice,as indicated by RT-PCR.Expression of3cytokines,IL-1?,IFN-?and TNF-?was all positive in the spleen tissue of both groups,suggesting that there was no significant difference in the levels of cytokine gene transcripts in both groups.Conclusion These findings suggest that murine resistance to central nervous system infection of C.neoformans be enhanced by intracerebral administration of H-CN,and anti-cryptococcal mechanism probably involves a local cytokine IL-1?elicitated by H-CN in central nerve system.

OBJECTIVEThe study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.

METHODBased on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.

RESULTUnder a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.

CONCLUSIONPRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.

Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 8 ; genetics ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Fas Ligand Protein ; metabolism ; Humans ; Magnoliopsida ; chemistry ; Rhizome ; chemistry ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; fas Receptor ; metabolism

Investigation of simvastatin and its related substances was carried out using a reversed phase ultra performance liquid chromatography/tandem mass spectrometry method. The identification of impurities in simvastatin was performed with a triple-quadrupole mass spectrometer, with an electrospray ionization (ESI) source in the negative/positive ion mode. A total of 12 compounds were characterized in commercial samples, among which 2 impurities had never been reported. All the impurities were deduced based on the MS fragment pathways of simvastatin and the biosynthetic pathway of lovastatin. This work provides very useful information for quality control of simvastatin.
Chromatography, High Pressure Liquid ; Chromatography, Reverse-Phase ; Drug Contamination ; Hypolipidemic Agents ; chemistry ; Quality Control ; Simvastatin ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Tandem Mass Spectrometry

To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid ; methods ; Ginkgolides ; analysis ; Injections ; Limit of Detection ; Salicylates ; analysis ; Tandem Mass Spectrometry ; methods

AIM:To detect the activation of macrophage autophagy caused by lipopolysaccharide ( LPS) and the possible related signaling pathways .METHODS:The macrophage cell line RAW264.7 cultured in vitro was divided into 5 groups according to the culture environment , including normal culture group , starvation-activated sautophagy group , LPS group, LPS+PI3K inhibitor (hVps34) group and LPS+mTOR inhibitor (rapamycin) group.Fluorescent expression vector pcDNA3.1-GFP-LC3 constructed in previous work was transfected into the macrophages .The fluorescence microscopy was used to detect the formation of autophagosome .The mRNA expression of autophagy-associated genes Atg5, Atg7, LC3-II and Bnip3 in the macrophages was detected by qRT-PCR.The protein levels of LC3-II, p-Akt and p-mTOR were deter-mined by Western blotting , so as to evaluate the molecular pathways of autophagy in LPS-activated macrophages .RE-SULTS:The macrophages stably expressing GFP-LC3 were successfully established , which were used to observe the auto-phagy under fluorescence microscope .Compared with normal culture group , the autophagy in starvation group , LPS +hVps34 group and LPS+rapamycin group was significantly increased .The mRNA expression levels of Atg5, LC3-II and Bnip3 were significantly increased in starvation group , LPS+hVps34 group and LPS +rapamycin group , while in LPS group, those decreased slightly .The protein level of p-Akt in starvation group , LPS group and LPS+rapamycin group was significantly increased , while p-mTOR in starvation group , LPS+hVps34 group and LPS+rapamycin group significantly declined .LC3-II expression level in starvation group , LPS+hVps34 group and LPS+rapamycin group was higher than that in control group and LPS group .CONCLUSION: LPS regulates macrophage autophagy , and its possible pathway is the PI3K/Akt/mTOR pathway, but there are some other effective regulatory pathways .

AIM: To evaluate the efficacy and safety of two different surgical techniques on congenital cataract on children.METHODS: Twenty-two children (1-3 years old) with congenital cataract were randomly divided into two groups (group A and group B). With group A (10 patients, 20 eyes), we applied 23-gauge (23G) trans corneal limbus vitrectomy system to complete lens cortex gettering, posterior capsulotomy and anterior vitrectomy；With group B (12 patients, 24 eyes), we used the phacoemulsification I/A to complete lens cortex gettering, and performed anterior vitrectomy with anterior vitreous cutting instrument. After that, the differences in intraoperative and postoperative complications between two groups were compared. RESULTS：In group A, the width of corneal limbal incision was 0.6mm, the incision was self-sealing, and the anterior chamber was stable and iris did not prolapse during the surgery. In group B, the width of corneal limbal incision was 3mm, anterior chamber was unstable and intraoperative iris prolapse occurred in 14 eyes (58%). And the incision need to be stitched up after surgery. In the postoperative follow-up of 6-24 months (an average of 14 months), we found that corneal neovascularization did not occur in group A, while in group B, corneal neovascularization occurred in four eyes (17%); Other complications, such as posterior capsular opacification，retinal detachment, glaucoma, hypotony or endophthalmitis did not occur in either group.CONCLUSION: The 23G trans corneal limbus vitrectomy system used in pediatric cataract surgery is safer and more effective than phacoemulsification I/A. It is promising in treatment of congenital cataract on children.

Objective To study the changes of phosphatidylcholine-specific phospholipase D (PC-PLD) enzyme activity of glomerular mesangial cells (GMCs) in high glucose and the effect of PLD on cytoskeleton. Methods Rats GMCs were cultured in high glucose(30 mmol/L) for 48 hours. PLD enzyme activity was measured by enzyme coupled colorimetry. PKC activity was measured by substrate phosphorylation, and fluorescence intensity of F-actin was observed by FITC-labeled antibody and laser scan cofocal microscopy (LSCM) . Results In high glucose, PLD and PKC activities were obviously elevated, but fluorescence intensity of F-actin was decreased and its cytoskeletal pattern was disassembly. After treatment with inhibtor of PLD, PKC activity was significantly decreased, and intensity and organization of F-actin were recovered. Conclusion Elevation of PLD activity induced by high glucose may influence the cytoskeleton and contraction of CMC through PKC pathway.

0.05).Mean time for lesion healing in immunocompromised and immunocompenent mice was 36.8 d and 29.0 d respectively(P0.05).Conclusions Primary cutaneous cryptococcosis may lead to dissemination of the infection in immunocompromised mice and our data suggest that skin is a possible route for dissemination of cryptococcal infection.