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

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.

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.

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 .

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

Adolescent ; Cholesteatoma ; complications ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans ; Male ; Temporal Bone

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

Evodiamine is one of the most important antitumor alkaloid from evodiamine. This study focused on the mechanism of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mammalian target of rapamycin (mTOR) signal pathway, in order to explore its antitumor mechanism and lay a foundation for clinical treatment of gastric cancer. The sole cytotoxic effect of evodiamine on SGC-7901 cells and human peripheral blood mononuclear cells (PBMCs) was observed by MTT assay. After the cells were respectively intervened with single evodiamine or evodiamine combined with z-VAD-fmk, the gene expressions of mTOR, p70S6K and 4EBP1 were analyzed by real-time PCR, and the protein expressions of mTOR and p-mTOR were detected by western blot. The result showed that evodiamine inhibited the apoptosis of SGC-7901 cells in a time-dependent manner, with no cytotoxic effect on human PBMCs. After the respective intervention with single evodiamine or evodiamine combined with z-VAD-fmk, the cells became round and floated in medium. Compared with the control group, both treatment methods can inhibit mTOR, 4E-BP1 and p70S6K gene expressions, with significant differences. Compared with single evodiamine, evodiamine combined with z-VAD-fmk showed a higher inhibitory rate in gene expression. According to the Western Blot result, evodiamine can inhibit the protein expressions of mTOR and p-mTOR regardless of the combination with z-VAD-fmk, with a higher inhibitory rate after z-VAD-fmk blocked caspase. In conclusion, evodiamine may promote the apoptosis of SGC-7901 cells through mTOR signal pathway.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Quinazolines ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; TOR Serine-Threonine Kinases ; genetics ; metabolism

Objective To evaluate the effect of sevoflurane postconditioning on microRNA-133a (miR-133a) expression during myocardial ischemia-reperfusion (I/R) in mice.Methods Thirty adult male C57 mice,weighing 20-30 g,were randomized to 3 groups (n =10 each) using a random number table:control group (group C),I/R group,and sevoflurane postconditioning group (group SP).In I/R and SP groups,hearts from adult male C57 mice were exposed and subjected to 30 min of ischemia and 180 min of reperfusion in anesthetized mice according to the method described by Das et al.In group C,only thoracotomy was performed without ligation of the coronary artery.In group SP,2.4％ sevoflurane was inhaled for 5 min starting from the onset of reperfusion to perform sevoflurane postconditioning.At 180 min of reperfusion,blood samples from the femoral vein were collected for determination of serum lactic dehydrogenase (LDH) and creatine kinase (CK) activities using the colorimetric method.The mice were then sacrificed,and myocardial specimens were obtained for determination of myocardial infarct size,miR133a and caspase-9 mRNA expression (by real-time reverse transcriptase polymerase chain reaction),and caspase-9 expression (by Western blot).Results Compared with group C,the serum LDH and CK activities and myocardial infarct size were significantly increased in I/R and SP groups,the expression of miR-133a was significantly down-regulated,and the expression of caspase-9 protein and mRNA was significantly up-regulated in group I/R,and the expression of miR-133a and caspase-9 protein and mRNA was significantly up-regulated in group SP (P＜0.05).Compared with group I/R,the serum LDH and.CK activities and myocardial infarct size were significantly decreased,the expression of miR-133a was significantly up-regulated,and the expression of caspase-9 protein and mRNA was significantly downregulated in group SP (P＜0.05).Conclusion The mechanism by which sevoflurane postconditioning inhibits cell apoptosis during myocardial I/R is related to up-regulation of miR-133a expression in mice.