Fluorescence decrease ratio was applied to determine of arte misinin （qinghaosu, QHS） based on the catalytic effect of hemoglobin （Hb） using tetraethyldiaminoxanthenyl chloride （Pryonine B, PB） as monitor in thi...Fluorescence decrease ratio was applied to determine of arte misinin （qinghaosu, QHS） based on the catalytic effect of hemoglobin （Hb） using tetraethyldiaminoxanthenyl chloride （Pryonine B, PB） as monitor in this work, Experimental results show that the catalytic characteristics of Hb for QHS, being expressed as Michaelis-Menten parameters, Km, Vmax and Kcat are 2.8×10^-5 mol·L^-1, 4.2×10^-5 mol·L^-1·s^-1 and 280 s^-1 , respectively. Like hemin, enzymatic bioactivities of Hb is inhibited by both deactivated agents and high temperature whereas enhanced by ethanol. With the catalytic action of Hb, quantitative method of QHS can be established based on the fluorescence decrease of PB and linear relationship between the fluorescence decrease ratio F0/F and the concentration of QHS is in the range of （0.0-1.1）×10^-6 mol·L^-1 with detection limit （3σ） being 7. 2×10^-9 mol·L^-1, The concentration of QHS in the media of plasma or urine was detected using this method.展开更多
Fluorescence decrease ratio （F0/F） was applied to determination of artemisinin （qinghaosu, QHS） based on the catalytic effect of tyrosinase using tetraethyldiaminoxanthenyl chloride （pyronine B, PB） as monitor. ...Fluorescence decrease ratio （F0/F） was applied to determination of artemisinin （qinghaosu, QHS） based on the catalytic effect of tyrosinase using tetraethyldiaminoxanthenyl chloride （pyronine B, PB） as monitor. A catalyst used commonly in the decomposition of QHS, tyrosinase, exhibited higher binding activity than hemin, which was expressed as Michaelis-Menten parameters, km, Vmax, and kcat respectively. Interaction of QHS with tyrosinase was inhibited in the presence of deactivating agents at high temperature whereas enhanced by ethanol. Under optimal conditions, a concentration of 1.4×10^-7-8.4×10^-7 mol·L^-1of QHS could be determined on the basis of fluorescence decrease ratio of PB, with a detection limit 3tr of 2.6×10^-9 mol·L^-1. The proposed method was applied to detection of the concentration of QHS in the media of plasma and urine.展开更多
DNA differential stain is a simple method distinguishing cells of proliferative from quiescent stage. Double stranded DNA in quiescent cells is easily denatured by weak acid into single strand. As double stranded nucl...DNA differential stain is a simple method distinguishing cells of proliferative from quiescent stage. Double stranded DNA in quiescent cells is easily denatured by weak acid into single strand. As double stranded nucleic acid combined with methyl green and single stranded nucleic acid with pyronin, we make use of methyl green pyronin staining method to the cells treated with weak acid to distinguish proliferating from quiescent cells. This paper reports the observation of leukemia cells in the bone marrow smears of 100 cases of untreated acute leukemia by DNA differential staining method. The percentage of Go cells was lowest in ALL and highest in APL.展开更多
[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathologica...[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathological sections of sick rabbits were stained and identified. [ Result] The pathological changes in brain tissue could be clearly observed on sections, but parasites were not examined in pathological brain tissues stained by common staining method. When the pathological section was stained by improved Gram staining method, the pathological changes in brain tissue were not ouly stained very clearly, but blue parasites were also found in brain tissues. The parasites in epithelioid cells were stained into purple ones by methyl green pyronin staining method. [ Conclusion] The im- proved Gram staining method and methyl green pyronin staining method performed good staining effects of E. cuniculi in pathological sections, which were conducive to rapid diagnosis of encephalitozoonosis in rabbit.展开更多
Elevated level of hypochlorous acid(HClO)is closely associated with cancer development.Identifying HClO level in cancer cells would provide important evidence in either early-stage cancer diagnostics or monitoring of ...Elevated level of hypochlorous acid(HClO)is closely associated with cancer development.Identifying HClO level in cancer cells would provide important evidence in either early-stage cancer diagnostics or monitoring of its treatment efficiency.In this work,a new pyronine-based fluorescent probe for rapid and sensitive detection of HClO was developed by condensing meso-formyl pyronine(PyCHO)with 2-hydrazinopyridine to form meso-pyridylhydrazone-functionalized pyronine PyHP,PyHP is nonfluorescent due to the excited-state C=N isomerization nonradiative decay,whereas the HClO-triggered formation of meso-triazolopyridyl pyronine PyTP abolishes the C=N isomerization and thus greatly enhances the fluorescence.With the probe,the cancer cells/tumor were distinguished with high-contrast from normal ones by laser confocal fluorescence imaging,and the tumor-to-normal(T/N)ratios obtained exceed the clinically acceptable threshold of 2.0.Moreover,its capability of in vivo imaging tumor was also demonstrated.These results indicate the potential of PyHP as an effective tool in the early clinical diagnosis of cancers.展开更多
基金Supported by the National Natural Science Foundation of China(20075012) the Scientific Research Foundation of Hunan Educational Commit-tee (02C313)
文摘Fluorescence decrease ratio was applied to determine of arte misinin （qinghaosu, QHS） based on the catalytic effect of hemoglobin （Hb） using tetraethyldiaminoxanthenyl chloride （Pryonine B, PB） as monitor in this work, Experimental results show that the catalytic characteristics of Hb for QHS, being expressed as Michaelis-Menten parameters, Km, Vmax and Kcat are 2.8×10^-5 mol·L^-1, 4.2×10^-5 mol·L^-1·s^-1 and 280 s^-1 , respectively. Like hemin, enzymatic bioactivities of Hb is inhibited by both deactivated agents and high temperature whereas enhanced by ethanol. With the catalytic action of Hb, quantitative method of QHS can be established based on the fluorescence decrease of PB and linear relationship between the fluorescence decrease ratio F0/F and the concentration of QHS is in the range of （0.0-1.1）×10^-6 mol·L^-1 with detection limit （3σ） being 7. 2×10^-9 mol·L^-1, The concentration of QHS in the media of plasma or urine was detected using this method.
基金Project supported by the National Natural Science Foundation of China （No. 20075012） and the Natural Science Foundation of Hunan Educational Committee （No. 02C313）.
文摘Fluorescence decrease ratio （F0/F） was applied to determination of artemisinin （qinghaosu, QHS） based on the catalytic effect of tyrosinase using tetraethyldiaminoxanthenyl chloride （pyronine B, PB） as monitor. A catalyst used commonly in the decomposition of QHS, tyrosinase, exhibited higher binding activity than hemin, which was expressed as Michaelis-Menten parameters, km, Vmax, and kcat respectively. Interaction of QHS with tyrosinase was inhibited in the presence of deactivating agents at high temperature whereas enhanced by ethanol. Under optimal conditions, a concentration of 1.4×10^-7-8.4×10^-7 mol·L^-1of QHS could be determined on the basis of fluorescence decrease ratio of PB, with a detection limit 3tr of 2.6×10^-9 mol·L^-1. The proposed method was applied to detection of the concentration of QHS in the media of plasma and urine.
文摘DNA differential stain is a simple method distinguishing cells of proliferative from quiescent stage. Double stranded DNA in quiescent cells is easily denatured by weak acid into single strand. As double stranded nucleic acid combined with methyl green and single stranded nucleic acid with pyronin, we make use of methyl green pyronin staining method to the cells treated with weak acid to distinguish proliferating from quiescent cells. This paper reports the observation of leukemia cells in the bone marrow smears of 100 cases of untreated acute leukemia by DNA differential staining method. The percentage of Go cells was lowest in ALL and highest in APL.
基金Supported by National Natural Science Foundation of China(31372407)
文摘[ Objective] The paper aimed to search new identification methods of Encephalitozoon cuniculi on tissue sections. [ Method] Using improved Gram staining method and methyl green pyronin staining method, the pathological sections of sick rabbits were stained and identified. [ Result] The pathological changes in brain tissue could be clearly observed on sections, but parasites were not examined in pathological brain tissues stained by common staining method. When the pathological section was stained by improved Gram staining method, the pathological changes in brain tissue were not ouly stained very clearly, but blue parasites were also found in brain tissues. The parasites in epithelioid cells were stained into purple ones by methyl green pyronin staining method. [ Conclusion] The im- proved Gram staining method and methyl green pyronin staining method performed good staining effects of E. cuniculi in pathological sections, which were conducive to rapid diagnosis of encephalitozoonosis in rabbit.
基金Scientific and Technological Innovation Program of Colleges and Universities in Shanxi Province(Nos.2021L529,2021L530)the Fund for Shanxi“1331 Project”。
文摘Elevated level of hypochlorous acid(HClO)is closely associated with cancer development.Identifying HClO level in cancer cells would provide important evidence in either early-stage cancer diagnostics or monitoring of its treatment efficiency.In this work,a new pyronine-based fluorescent probe for rapid and sensitive detection of HClO was developed by condensing meso-formyl pyronine(PyCHO)with 2-hydrazinopyridine to form meso-pyridylhydrazone-functionalized pyronine PyHP,PyHP is nonfluorescent due to the excited-state C=N isomerization nonradiative decay,whereas the HClO-triggered formation of meso-triazolopyridyl pyronine PyTP abolishes the C=N isomerization and thus greatly enhances the fluorescence.With the probe,the cancer cells/tumor were distinguished with high-contrast from normal ones by laser confocal fluorescence imaging,and the tumor-to-normal(T/N)ratios obtained exceed the clinically acceptable threshold of 2.0.Moreover,its capability of in vivo imaging tumor was also demonstrated.These results indicate the potential of PyHP as an effective tool in the early clinical diagnosis of cancers.
