1.An albino strain of Aspergillus fumigatus causes pulmonary aspergilloma in an individual with normal immune ;system
Fang LIU ; Qingtao KONG ; Caiyun ZHANG ; Hong SANG ; Yongnian SHEN ; Guixia LYU ; Weida LIU ; Yi SHI
Chinese Journal of Dermatology 2016;49(8):563-567
A 76?year?old female patient complained of right chest pain for three months. CT scan showed a clump?like high?density shadow measuring 4.8 cm × 3.0 cm in size in the dorsal portion of the right lower lobe of the lung. Aspiration biopsy was performed, and biopsy samples were subjected to fungal culture and histopathological examination. Histopathological examination showed chronic granulomatous inflammation with hyaline septate hyphae. After 4?day culture, white villous dense colonies were formed on the Sabouraud′s agar medium. The center of the colonies was slightly elevated with wrinkles or radiating striae on the surface, and the bottom of the colonies was faint yellow in color. Microculture yielded abundant septate branched hyphae, and very few colorless hyaline quasi?circular spores. DNA sequencing of rDNA internal transcribed spacer (ITS) regions and β?tubulin genes was performed to identify the isolate, and antifungal susceptibility testing was carried out in vitro. The MEGA7.0 software was used to build phylogenetic trees of Aspergillus fumigatus complex and its closely related species. The isolate was identified as Aspergillus fumigatus by molecular biologic sequencing. The patient was diagnosed with pulmonary aspergilloma. After administration of itraconazole oral solution and vorionazole tablets, the condition got better obviously.
2.GC-MS Analysis of Volatile Oil from Chaihu Guizhi Decoction and Study on Its Inhibitory Effects on in vitro Proliferation of Human Lung Adenocarcinoma A 549 Cells
Ran WEN ; Zhuangzhuang LI ; Yiqing DU ; Yong YANG ; Rong RONG ; Qingtao LYU
China Pharmacy 2021;32(1):29-33
OBJECTIVE:To study the compositi on of the volatile oil from Compound chaihu guizhi decoction ,and to evaluate its in vitro anti-proliferative activity on human lung adenocarcinoma A 549 cells. METHODS :The volatile oil from Chaihu guizhi decoction was extracted according to the steam distillation method of general rules 2004 in the 2015 edition of Chinese Pharmacopoeia(part Ⅳ). The volatile oil components were analyzed by GC-MS combined with Kováts index ,and the relative content of each component was calculated by peak area normalization method. Using different concentrations of cisplatin (4,8, 16,32,64 mg/L)as positive control ,MTT assay was used to detect the inhibitory effects of different concentrations of volatile oil from Chaihu guizhi decoction (25,50,100,200,400 mg/L)on in vitro proliferation of A 549 cell after 48 h of treatment. Negative control group (with cells but without drugs )was set up. RESULTS :A total of 71 chemical components were isolated from the volatile oil ,among which there were 59 compounds identified ,sum of peak areas accounting for 84.99% of the total peak area. The compounds with relatively high content included ar-curcumene (17.65%),β-bisabolene(9.57%),β-ocimene(7.05%), α-curcumene(5.35%),2,5-dimethylbenzaldehyde(4.24%),linalyl isobutyrate (2.70%),α-cedrene(2.48%),δ-cadinene (2.07%). Compared with negative control group ,the proliferation rate of cells were decreased significantly in 4-64 mg/L cisplatin groups and 25-400 mg/L volatile oil from Chaihu guizhi decoction groups (P<0.05). IC 50 of cisplatin and volatile oil from Chaihu guizhi decoction to in vitro proliferation of A 549 cells were 10.150 and 73.526 mg/L. CONCLUSIONS :The volatile oil from Chaihu guizhi decoction mainly includes ar-curcumene ,β-bisabolene,β-ocimene,α-curcumene,which shows certain inhibitory effect on in vitro proliferation of A 549 cells.
3.Establishment of a candidate reference measurement procedure for the enumeration of cell particles in urine and applied to multi-center evaluation of an automated urine analyzer
Yuhong YUE ; Zhiqi GAO ; Ping WANG ; Yan QIAO ; Changjun LYU ; Jie LIU ; Liyan CUI ; Yongtong CAO ; Qingtao WANG
Chinese Journal of Laboratory Medicine 2022;45(4):388-392
Objective:To establish a candidate reference procedure for the enumeration of cell particles in urine and applied to the multi-center performance evaluation of an automated urine formed elements analyzer.Methods:According to the standardized mannual microscopic examination of fresh non-centrifuged urine samples and the recommended reference method for enumeration of cell particles in urine published by ISLH, we established a candidate reference procedure for the enumeration of cell particles in urine. From four class A tertiary hospitals′ clinical laboratories, three rigorous trained technicians per hospital tested the same specimen respectively using the reference procedure. Each specimen was repeatedly counted 5 times, obtaining the quantitative results of cell particles were obtained in urine. Four hospitals used the established candidate reference measurement procedure and the automated urine formed elements analyzer to detect 40 to 60 urine specimens from September 2020 to January 2021, and evaluate the established reference method, meanwhile evaluate the accuracy and consistency of the each count from automated urinalysis analyzer.Results:Using the candidate reference measurement procedures, the coefficient of variation of results derived from three trained technicians per hospital was less than 6.98% (red blood cells), 6.99% (white blood cells), 13.94% (epithelial cells) and met the quality requirements. The performance evaluation results of automated urine formed elements analyzer showed that the accuracy of red blood cells, white blood cells and epithelial cells met the requirements (bias≤4.98%) and was well consistent with the reference measurement procedure ( R2≥0.989). Conclusions:A candidate reference measurement procedure for the enumeration of urine cell particles was successfully established with satisfactory precision and accuracy. This procedure was applied to multicenter performance evaluation of an automated urine formed elements analyzer with good accuracy and consistency.
4.Content Determination of 8 Components in Astragalus membranaceus before and after Bidirectional Solid Fer- mentation with Cordyceps kyushuensis by HPLC-DAD-ELSD
Yiqing DU ; Qinhui YU ; Ran WEN ; Tongtong JIAN ; Rong RONG ; Qingtao LYU ; Guoying ZHANG
China Pharmacy 2020;31(23):2841-2846
OBJECTIVE:To establish a method for simultaneous determination of calycosin glucoside ,ononin,calycosin, formononetin,astragaloside Ⅳ,isoastragaloside Ⅱ,cycloastragenol and isoastragaloside Ⅰ in Astragalus membranaceus before and after bidirectional solid fermentation with Cordyceps kyushuensis ,and to investigate the effects of fermentation on the contents of above 8 components in A. membranaceus . METHODS :HPLC-DAD-ELSD was adopted. The determination was performed on Agilent 5 TC-C18 column with mobile phase consisted of 0.1% formic acid aqueous solution-acetonitrile (gradient elution )at the flow rate of 1 mL/min. The column temperature was set at 30 ℃. DAD detection wavelength was set at 260 nm,ELSD evaporation tube temperature was 100 ℃,atomizer temperature was 80 ℃,carrier gas flow rate was 1.6 L/min;injection volume was 15 μL. RESULTS:The eight components had a good linear relationship within their respective ranges of concentration (all R2>0.999 0); RSDs of precision ,stability and repeatability tests were all less than 3%(n=3 or n=6);the recoveries was 97.88%-101.32%, and RSDs were 1.22%-2.39%(n=6). Setting the content of components in unfermented A. membranaceus as 100%,after bidirectional solid fermentation with C. kyushuensis ,the change rates of 8 components were -98.51%,-96.41%,-94.74%, -96.40%,289.20%,20.25%,-75.05%,562.46%,respectively. CONCLUSIONS :After fermentation with C. kyushuensis ,the contents of active components as astragaloside Ⅳ,isoastragaloside Ⅰ and isoastragaloside Ⅱ can be increased significantly in A. membranaceus .