1.The genetic polymorphisms of mannose binding lectin and Kawasaki disease
International Journal of Pediatrics 2012;39(3):217-219
Kawasaki disease(KD) is a common autoimmune disease in children.KD is characterized as a systemic artcriolar inflammatory,but its pathogenesis is still not clear.In recent years,many studies have shown KD is associated with the polymorphisms of multiple genetic fragments.The relationship between mannose binding lectin(MBL) gene and KD has attracted much attention.This review introduces the construction and function of MBL briefly,and focuses on the relationship between the genetic polymorphisms of MBL and susceptibility,cardiovascular damage of KD.
2.Establishment of a quantitative method for leukocyte fragments in apheresis platelet concentrates and its influential factors
Guangming JIANG ; Linjie ZHANG ; Baolong WANG ; Shulai YE ; Jiaping CHEN ; Jianhua ZHOU ; Min WANG ; Yanqiu LIAO
Chinese Journal of Laboratory Medicine 2009;32(7):798-804
Objective To establish a new method for quantitating leukocyte fragments (LFs) in apheresis platelet concentrates (AP-PCs) by using real-time quantitative polymerase chain reaction (RQ-PCR) and flow cytometry(FCM) and discuss the factors influencing LFs concentrations such as storage time, filtration and PLT concentration. Methods 67 qualified donors were selected. Each of them donated one therapeutic dose of AP-PCs. AP-PCs samples were collected as soon as possible and divided into si xfractions. One was analyzed by hematology analyzer. For the Others, DNA was extracted under differen tconditions (filtrated or unfiltrated, before or after centrifugation) at 4 hours, 24 hours, 48 hours, 72 hours, 96 hours after blood draw, respectively. Then the amounts of albumin gene of the AP-PCs and the cell-free DNA in supematant were quantitatively determined using RQ-PCR and the results were calculated into leukocytes equivalent(WBCs/μl). Intact leucocytes were counted by FCM. The concentrations of LFs were calculated by subtracting cell-frce DNA and intact leucocytes from the total DNA amount. Then the differences of LFs concentrations among groups with different storage time were compared and the differences of LFs concentrations between unfihrated and filtrated groups were also compared. After grouping all the AP-PCs according to their PLT concentrations, LFs contents of AP-PCs before filtration among groups were compared. Meanwhile, bivariate correlation analysis between PLT concentrations and LFs contents was carried out. ResultsLFs contents of all the AP-PCs samples were quantitated successfully.The concentrations of LFs in AP-PCs before filtration in 4 hours,24 hours,48 hours,72 hours , 96 houres after blood draw were(31.4±17. 6), (47.5±25.3), (100.7±53.5), (89.5 ±47.2) and (16.1±7.8) WBCs/μl ; After filtration the results were (16. 9±8. 7), (24. 3 ± 12. 2), (83. 1±42. 6), (78.2 ±40. 2) and (13.6 ± 6. 6) WBCs/μl respectively. There were statistically significant differences among groups of different storage time (Fwithin subjects = 472. 756,P < 0.01). The concentrations of LFs kept on increasing within 48 hours after collections, and then decreased gradually. The peaks appeared between 48 hours and 72 hours after collections. The differences of LFs contents between unfiltered and filtered AP-PCs in 4 hours, 24 hours, 48 hours, 72 hours, 96 hours after collections were 14. 5, 23. 2, 17. 6, 11.3 and 2. 5 WBCs/μl, respectively.There was statistically significant difference between unfiltered and filtered samples (Fbetween subjects=9. 216,P < 0. 05). The differences were considerable within 48 hours, and then declined gradually. The results of bivariate correlation analysis showed that there were no statistically significant correlation between PLT concentrations and LFs contents (at 4, 24, 48, 72, 96 hours after collections the correlation coefficients rs were -0.002, 0.015, 0.027, 0.042 and 0. 037,respectively,P2-tailed>0.05). ConclusionsRQ-PCR and FCM can be used to quantitate LFs in AP-PCs. The concentration of LFs in AP-PCs is influenced by storage time and filtration, but it is not affected by PLT concentration.
3.Determination of the raw material feeding (Gentiana scabra, Katsumade Galangal Seed and dried tangerine peel) in compound gentian tincture by HPLC-MS/MS
Chao LIU ; Xiaolin WANG ; Fudong ZHANG ; Shulai ZHOU ; Yuzhu LIU
International Journal of Traditional Chinese Medicine 2018;40(9):854-860
Objective To establish a HPLC-MS/MS method for comprehensive monitor and control of the raw material feeding (Gentiana scabra,Katsumade Galangal Seed and dried tangerine peel),and determination of Gentiopicroside,Alpinetin,Cardamonin and Hesperidin in Compound gentian tincture.Methods The separation was performed on an Inertsil ODS-3 (4.6 mm× 150 mm,5 μm) analytical column with the mobile phase consisting of acetonitrile-0.1% formic acid solution by gradient elution program,and the column temperature was 40 ℃.Active ingredients were separated by HPLC.The Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode,and reactions ion monitoring mode (MRM) for quantitative analysis were selected.Results Through the analysis of the samples with mixed extract the same characteristic peak in MS was found to determine the proprietary Chinese medicine according to the prescription feeding process.The calibration curve of Gentiopicroside,Alpinetin,Cardamonin and Hesperidin were linear:103.26-619.56 μg (r=0.999 0),109.50-657.00 μg (r=0.999 5),105.50-633.00 μg (r=0.996 9),105.02-630.12 μg (r=0.999 5).The precision was Gentiopicroside 0.81%,Alpinetin 0.48%,Cardamonin 0.61% and Hesperidin 1.55% respectively.The average recovery rate were Gentiopicroside 95.08%,Alpinetin 93.28%,Cardamonin 94.78% and Hesperidin 95.04% respectively.Conclusions The method was proved to be simple,accurate,reliable,high sensitivity and can be used for determination and control of the raw material feeding (Gentiana scabra,Katsumade Galangal Seed and dried tangerine peel).