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.
3.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.
4.Suitable Hospital Infection Control Measures in Health Centers of Poverty-striken Villages
Yimin GU ; Jiahui GU ; Hongyan JI ; Yao SUO ; Shuai YANG
Chinese Journal of Nosocomiology 1994;0(04):-
OBJECTIVE To explore the suitable hospital infection control measures in health centers of poverty-striken villages,in order to improve the management of hospital infection,decrease hospital infection rate and protect the health of medical staff and patients.METHODS The status quo of hospital infection in health centers of poverty-striken villages,was investigated in 20 small towns health centers with were randomly divided into two groups:test group(n=15)and control group(n=5).The suitable hospital infection control measures were explored from 5 points.The effect of infection control by before-after controlled study of experimental group and randomized controlled study of control group was anal yzed.RESULTS The rate of hospital infection in test group was decreased from 7.60% to 1.98% and at in control group didn't change,the difference was significant.CONCLUSIONS The managements of establishment of the suitable hospital infection control measures in health centers of poverty-striken villages have been put into practice and gained good result.
5.Determination of Nine Aconitum Alkaloids in Xiaohuoluo Pills with Quantitative Analysis of Multi-compo-nents by Single Marker
Hongyan CHEN ; Xiao SUN ; Xiaolong ZHANG ; Chenxu YAO ; Yan JIANG
China Pharmacist 2017;20(9):1545-1549
Objective:To develop a method of quantitative analysis of multi-components by single marker( QAMS) for nine kinds of alkaloids in Xiaohuoluo pills. Methods: An HPLC-QTOF-MS method with an Agilent ZORBAX Extend-C18 RRHT(2. 1 mm × 50 mm,1. 8 μm) column was applied. The flow rate was 0. 21 ml·min-1 . The column temperature was 30 ℃. The mobile phase was methanol (A)-water (B;containing 0. 1% formic acid and 2. 5 mmol·L-1 ammonium acetate) with gradient elution. The aconitine was used as the internal standard, and the relative correction factor ( RCFs) of hypaconitine, mesaconitine, benzoylaconine, benzoyl-hypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine was respectively established, and the reproducibility inspection on the RCF was performed. The contents of the other 8 kinds of aconitum alkaloids were calculated according to the RCF. At the same time, an external standard method ( ESM) was performed for the content determination of the nine alkaloids. The results of the two methods were compared. The feasibility and accuracy of the QAMS method were verified. Results:Within a certain range,the RCF of hypacontine,mesacontine, benzoylaconine, benzoylhypaconine, benzoyl mesaconine, aconine, hypaconine and mesaconine to aconitine was 1. 736,1. 979,1. 0471,0. 9242,1. 2901,1. 3078,1. 2859,and 1. 0948,respectively. The QAMS method was established for determi-ning alkaloids. There were no significant differences between the results of the QAMS method and those of the external standard method ( ESM) . Conclusion:With the validation of methodology, the method established in our study can be used for the content determina-tion of aconitine, hypaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, benzoylmesaconine, aconine, hypaconine and mesaconine in xiaohuoluo pills.
6.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.
7.Initial experimental research on placenta extract to stimulate lymphocyte proliferation activity and stability
Guohua WANG ; Wangtai CHENG ; Hongyan MA ; Zhiyang CHEN ; Libo YAO
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):137-139
Purpose The aim is to prepare the extract of immunocompetent cell proliferation from human placenta and to try to find out a suitable method of preserving the extract.Methods The extract was mainly prepared by heat-treating placenta homogenated fluid. Then the activity to stimulate murine splenic lymphocyte proliferation in vitro was done with MTT , exposing the extract to radioisotope 60 Co.Results The content of protein was 2~3mg per gram placenta measured by Lowry′s Method.The rate to promote cell proliferation was more than 80 percent.The activity lasted a few months after being exposed to radioisotope.Conclusion The extract prepared by heat-treating not only had high activity, but also had the unique method and good repeatability.This prepared extract as a kind of stable,reliable and remarkably promoting lymphocyte proliferation reaction had the value of development production on broad scale as well as in clinical practice.
8.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.
9.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.
10.Characteristics of the Body Constitution and Syndrome Type of Traditional Chinese Medicine (TCM) in Primary Sj(o)gren's Syndrome (pSS)
Hongyan SUN ; Muzhi CHEN ; Ankun LYU ; Hua YAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2016;18(12):2148-2153
This study aimed at exploring the features of the body constitution and syndrome types of TCM in patients with pSS.General information of 60 patients with pSS was collected.Their TCM syndrome types and TCM body constitution types were examed,while the distribution of pSS syndrome types in terms of their body constitution was analyzed.Among 60 pSS patients,58 of them (96.67%) were female with the average age of 52 years and an average course of 63 months.Their usual residence was Xinjiang.Their most common first symptoms included thirst,dry eyes and joint pain,and the damage of skin and mucous membranes and blood system was rare.It was found that the syndrome of both qi and yin deficiency was the most common (48.33%),followed by the impregnation syndrome of heat and dampness (26.67%) and the syndrome of the weakness of spleen qi (25%) in pSS patients.Yin deficiency (58.33%) and yang deficiency (58.33%) in the body constitution of TCM accounted for decent proportions,followed by the constitutions of qi stagnation (48.33%),qi deficiency (45.00%),blood stasis (38.33%),phlegm-dampness (36.67%),damp-heat (18.33%),allergic (13.33%) and yin-yang harmony (1.67%).Those without the health education of pSS accounted for 51.11% in cases of palindromia.In conclusion,yin deficiency and yang deficiency constitutions were most common,at most,the body constitution of both qi and yin deficiency,in pSS patients in accordance with TCM body constitution.Most pSS patients of palindromia didn't received professional health education of pSS.All the causes above suggested that we should make individualized treatment programs according to clinical syndrome types and the characteristics of TCM body constitution in pSS patients for strengthening the health management of pSS and improving their quality of life.