1.Determination of 7 Kinds of Ginsenoside in Qipi Pills by HPLC-QTOF-MS
Hongyan CHEN ; Chenxu YAO ; Xiaolong ZHANG
China Pharmacist 2017;20(3):460-462
Objective:To simultaneously determine the contents of ginsenoside Rb1 , ginsenoside Rb2 , ginsenoside Rb3 , ginsen-oside Re, ginsenoside Rg1 , ginsenoside Rf and ginsenoside Ro in Qipi pills by HPLC-QTOF-MS. Methods: The determination was performed on an Agilent Poroshell 120 EC-C18 column (2. 1 mm × 50 mm,2. 7 mm) with mobile phase consisting of acetonitrile( A)-water(B, containing 0. 1% formic acid) with gradient elution. The flow rate was 0. 21 ml·min-1. The column temperature was 30℃. The MS instrument was equipped with an ESI+ ion source. The exacted ion chromatograms were used to determine the quantities of different compounds in the samples while the mass spectra of product ions were used for confirming the compounds. Results:All the 7 kinds of ginsenoside showed good linearity (r>0. 9993). The RSDs of precision, repeatability and stability tests were all less than 5%. The average recoveries were within the range of 97. 11%-101. 98%. Conclusion:The method is simple, rapid and reliable with high specificity, which can be used for the quality control of Qipi pills.
2.Features and Pathogenesis of Subcortical Aphasia (review)
Jingfan YAO ; Hongyan CHEN ; Yumei ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2015;21(2):130-133
With the development of imaging techniques, aphasia caused by subcortical impairment is increasingly found in recent years. This paper reviewed the literature about the subcortical aphasia and expolred the anatomical and clinical features and pathogenesis of subcortical aphasia.
3.Expression and identification of recombinant Clostridium difficile toxin B using Bacillus megaterium system
Guilin YANG ; Weilong LIU ; Hongyan YAO ; Boping ZHOU ; Hanping FENG
Chinese Journal of Infectious Diseases 2011;29(1):1-5
Objective To express and purify recombinant and biologically active Clostridium difficile toxin B (rTcdB). Methods The genes of TcdB were amplified by polymerase chain reaction (PCR) using chromosomal DNA from a toxigenic strain, and cloned into a shuttle vector pHis1522.The sequences of TcdB genes in the vector were verified by DNA sequencing. The construction was transformed into Bacillus megaterium protoplasts and the protein expression was driven by a xylose promoter. The purified protein was tested for biological activity. Results rTcdB was successfully purified from bacterial crude extracts. Approximately 5-10 mg of highly purified recombinant toxin was obtained from one liter of bacterial culture. The expressed rTcdB had molecular mass similar to the native toxin, and its biological activity was proved to be similar to its native counterpart after an extensive examination. Conclusion rTcdB with biological activities is successfully expressed in Bacillus megaterium.
4.The differences of adverse drug reaction between children and adults caused by Tanreqing injection
Juan XU ; Hongyan WU ; Linqing YAO ; Xin ZHANG
International Journal of Traditional Chinese Medicine 2013;(2):106-108
Objective To analyze the differences of adverse drug reactions between children and adults caused by Tanreqing injection,and provide reasonable application of Tanreqing injection.Methods Retrospective research method were adopted to screen out the adverse drug reaction reports caused by Tanreqing injection from drug ADR reports of Drug ADR Monitoring Center affiliated to the Second People's Hospital of Gansu Province from January 2010 to October 2012.Figures of children and adults who used Tanreqing injection were analyzed separately.Results A total of 603 children used Tanreqing injection,among which 5 children appeared ADR (0.83%).A total of 4395 adult used of Tanreqing injection,among which 4 appeared in ADR (0.09%).There was statistical difference between the two groups (x2 =16.07,P< 0.05) The incidence rate of ADR in males was more than females either in the adults or in children.Hypersensitivity was the most common ADR.Conclusion ADR of Tanreqing injection in Children was significantly higher than adults.Tanreqing injection should be used carefully and rationally.
5.Current status of health-related productivity loss and its risk factors in nurses
Fang YANG ; Yan YAO ; Hongyan LI ; Jianbo SI ; Wei SONG
Chinese Journal of Health Management 2012;(6):405-408
Objective To explore current status of health-related productivity loss and its risk factors among nurses.Methods Stanford presenteeism scale (SPS-6) and self-designed questionnaire were used to investigate current status of health-related productivity loss and its risk factors among 1122 nurses working in a tertiary hospital in Changchun city.Results Compared with hired nurses,age and work seniority of permanent nurses were significantly higher(Z =-19.49,-19.28 ;P <0.05).The average score of SPS-6 of all the participants was 20.05 ± 4.37.The score of SPS-6 of married nurses was significantly lower than other nurses (Z =-3.52,P < 0.05) ; and the score of SPS-6 of nurses less than 30 years old was significantly higher than those above 30 years old (Z =-2.49,P < 0.05).There were no significant differences between the SPS-6 score of education degree and department.(Z =-1.37,x2 =0.58 ; P >0.05).The result of GLM showed that employment status and work seniority were independent risk factors of health-related productivity loss among nurses.The scores of SPS-6 of permanent nurses was significantly lower than hired nursed (x2 =4.48,P < 0.05),and those who had worked for less than 3 years showed significantly higher score of SPS-6 than those who had worked longer (x2 =12.89,P < 0.05).Conclusions Health-related productivity loss do exist among nurses.Improving health management may help to reduce this loss of productivity.
6.Preparation and identification of the monoclonal antibody against human bone sialoprotein
Hongyan DU ; Ligang JIE ; Xiaoyan YAO ; Ming LI
Cancer Research and Clinic 2011;23(2):81-84
Objective To prepare the human bone sialoprotein (BSP) monoclonal antibodies (mAb)with high titer and specificity and identify its characterization,which is based on further studying BSP as clinical biomarker for breast cancer metastasizing to bone. Methods BALB/c mice were immunized with purified recombinant BSP protein.Cell fusion was performed between mouse splenic cells and myeloma cells (Sp2/0), and then the hybridoma cell lines secreting mAb against BSP antigen were screened and cloned. The ascites were prepared and purified with Protein G affinity chromatography.The titer and subtypes of mAb against BSP were identified and measured by ELISA and Western blotting analysis. ResultsNine hybridoma cell lines that stably secreted mAb against BSP were successfully obtained.Two of them,D001 and D002,were further identified, which belonged to the subtypes of IgG1 and κ light chain. The two antibodies titers in culture supernatant were 1∶5120 and 1∶10 240, respectively, and those in the ascites fluid were 1∶25 600 and 1∶51 200,respectively.Results of Western blotting analysis and immunohistochemistry showed that the two antibodies could specifically bind with BSP derived from human breast cancer cells.ConclusionNine mAb against BSP have been successfully prepared which can be used for further studying the biological properties of BSP and reveal its relationship with data from clinic patients.
7.Determination of Alcohol Amine-diterpene Alkaloids by Hydrolysis Rule of Diester Aconitum Alkaloids
Xiaolong ZHANG ; Hongyan CHEN ; Xiao SUN ; Chenxu YAO ; Yan JIANG
China Pharmacist 2017;20(6):994-997
Objective: To investigate the hydrolysis conversion rate of alcohol amine-diterpene alkaloids from aconitum alkaloids, hydrolyze aconitum alkaloids reference substance, calculate the amount of alcohol amine-diterpene alkaloids in the hydrolysis solution by the hydrolysis conversion rate, which is used as the amount of alcohol amine-diterpene alkaloids reference substance, and establish a content determination method for aconine, hypaconitine and aconine in Aconiti radix cocta.Methods: Through controlling the hydrolysis conditions of aconitine, hypaconitine and mesaconitine, aconine, hypaconitine and aconine were obtained.The determination was performed on an Agilent ZORBAX Extend-C18 RRHT(2.1 mm×50 mm,1.8 μm) column with the mobile phase consisting of methanol(A)-water(B containing 0.1% formic acid and 2.5 mmol·L-1 ammonium acetate) with gradient elution by HPLC-QTOF-MS.The flow rate was 0.21 ml·min-1.The column temperature was 30 ℃.MS instrument was equipped with an ESI+ ion source.Results: Under the hydrolysis conditions of this study, the conversion rate of aconine from aconitine was 99.64%;the conversion rate of hypaconitine from hypaconine was 99.94%;the conversion rate of mesaconitine from mesaconine was 99.57%.The HPLC-QTOF-MS methodological investigation showed the 3 kinds of alcohol amine-diterpene alkaloids were with good linearity (r>0.999 1).The RSD of the precision, repeatability and stability tests were less than 5%.The average recoveries were within the range of 99.43%-100.10%.Conclusion: The validated method is simple, specific, reliable and reproducible.In the absence of reference substance, it can be used for the quality control of the herbs of Aconitum L.species.
8.Inactivation of HIV-1 in whole blood by methylene blue photochemical method
Yao ZHANG ; Hongyan XIONG ; Fei MA ; Zuoy BAO
Journal of Third Military Medical University 2003;0(21):-
Objective To observe the ability of inactivating human immunodeficiency virus type 1 (HIV-1) in whole blood by visible light combination with methylene blue (MB) at different concentrations. Methods HIV-1 was used as the test virus. The contaminated blood was treated by MB, visible light (640 nm), combined with MB and visible light (640 nm). The test of MT4 cell infection was used to evaluate the virus inactivation efficacy. Results After being treated by MB at the concentrations of 5, 10, and 15 ?mol/L, and then irradiated respectively by visible light (40 000 Lux, 640 nm) for 30, 20, and 10 min, all the added indicated virus at the titer of 10 5.78 TCID 50 of HIV-1 could be inactivated absolutely. Conclusion Methylene blue photochemical method can inactivate HIV-1 in blood effectively.
9.Effects of estrogen on behavior and expression of 5-HT in periaqueductal gray of migraine rats
Hongyan ZHANG ; Tingmin YU ; Xijing MAO ; Gang YAO
Journal of Jilin University(Medicine Edition) 2006;0(01):-
Objective To observe the effects of estrogen on behavior and 5-HT in periaqueductal gray (PAG)in migraine model rats. Methods Tewnty-four ovariectomized Wistar rats were divided randomly into four groups:control group (Group A),migraine group(Group B),low dose estradiol-treated ovariectomized group(Group C),high dose etradiol-treated ovariectomized group(Group D).After 1 week,the rats in Group B,C and D were injected with nitroglycerine 10 mg?kg-1 subcutaneously to make migraine rat models,the rats in Group A were given peanut oil alike,and the behavior changes were observed.2 h after injection,the rats were killed and the midbrains were separated and then 5-HT immunohistochemical staining was performed.Results Behavior: compared with Group B,the degrees of red-calws,red-ears and red-tail rats in Group D relieved obviously,the times of climbing hutch and scratching head were much fewer,while the rats in group C showed no significant difference;Immunohistochemical staining:compared with Group A,the 5-HT-positive neurons expression in PAG of Group B and C were more obviously(P
10.Effects of matrine on proliferation, cell cycle and apoptosis of human hepatocarcinoma cell line HepG2
Dandan ZHU ; Shukun YAO ; Jianguo YAN ; Hongyan LI
Cancer Research and Clinic 2010;22(11):745-747,751
Objective To study the effects of matrine on proliferation, cell cycle and apoptosis of human hepatocarcinoma cell line HepG2 and probe into the mechanisms of its anti-hepatocarcinoma effects.Methods The HepG2 cells were treated with different concentration of matrine (0.0125, 0.02, 0.05, 0.1, 0.2,0.4, 0.8, 1.6 g/L) for different time (24, 48, 72 h), then investigate the effects of matrine on cell proliferation by MTT, and the effects on cell cycle and apoptosis by flow cytometry. Results Matrine can inhibit the HepG2 cells proliferation at the concentration of 0.1 g/L and above in a concentration-dependent and timedependent manner(P <0.01). The result of FCM showed that the cell cycle of HepG2 was retarded at G1 phase treated with matrine for 48 h at the concentration of 0.8 g/L [(75.3±6.5)% vs (64.1±6.3)%, P <0.05], whereas was retarded at G2 phase treated with matrine for 48 h at the concentration of 1.6 g/L [(29.1 ±9.1)% vs (11.6±2.1)%, P <0.01]. The apoptosis of HepG2 cells can be induced by matrine for 12, 24 h or 48 h at the concentration of 0.4, 0.8, 1.6 g/L. Conclusion Matrine can inhibit cell proliferation, interfere cell cycle and inducing apoptosis of hepatocarcinoma cells, which may be involved in the mechanisms of matrine's antihepatocarcinoma effects.