2.Determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials by LC-MS/MS.
Hai-hong SI ; Yan-jing LI ; Jia XUE ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(14):2832-2836
To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid
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methods
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Ginkgolides
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analysis
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Injections
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Limit of Detection
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Salicylates
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analysis
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Tandem Mass Spectrometry
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methods
3.Application of circular dichroism to the study of interactions between small molecular compounds and DNA.
Zhen-jia LIU ; Yi-kang SI ; Xiao-guang CHEN
Acta Pharmaceutica Sinica 2010;45(12):1478-1484
Circular dichroism (CD) is an useful technique for monitoring DNA conformation changes resulting from changes in environmental conditions, such as temperature, ionic strength, and pH, and also for the study of the interaction between DNA and ligands (including small molecules and proteins). CD spectroscopy of DNA arises from the asymmetric backbone sugars and by the helical structures often adopted by nucleic acids. By the interpretation of induced circular dichroism (ICD) of ligand signals resulting from the coupling of electric transition moments of the ligand, DNA bases within the asymmetric DNA environment, ligand-DNA interactions, as well as the DNA-binding mode can be assessed. A number of important conclusions have been reported that related to the observed ICD signals resulting from the interactions between intercalators and groove binders with DNA. If short oligonucleotide sequences are used in the study, sequences-specific of binding also can be deduced. CD determination requires smaller amounts of sample, and not limited by the molecular weight or size and can be performed rapidly; though CD is of low resolution, but it's a complement to NMR and X-ray diffraction methods. This review will introduce the characters of the CD spectra of DNA, and its application to the studies of DNA with small molecules; some progress of the studies in our laboratory will also be discussed. CD is expected to be used as a screening method in seeking more DNA-targeted drugs, such as, antineoplastic, antimicrobial and antiviral drugs.
Animals
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Antineoplastic Agents
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chemistry
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Base Sequence
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Circular Dichroism
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methods
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DNA
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chemistry
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metabolism
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Humans
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Intercalating Agents
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chemistry
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Ligands
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Protein Binding
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Small Molecule Libraries
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pharmacology
4.Simultaneous determination of the four effective components in Huaijiao pill by HPLC.
Qing-Quan BIAN ; Zhen-Ping YANG ; Jia-Qin LIU ; Si-Man LIU
China Journal of Chinese Materia Medica 2005;30(19):1513-1515
OBJECTIVETo develop an HPLC method for determination of the four effective components (genistin; rutin; quercetin; genistein) in Huaijiao pill.
METHODThe chromatographic separation was performed on a Shim-packODS (4.6 mm x 150 mm, 5.0 microm) eluted with a mobile phase of MeOH-H2O-HAc (40:60:0.25). The detection wavelength was set at 256 nm and column temperature was set at 30 degrees C .
RESULTNice linear relation between the peak area and injected amount exists when the amount is within 0.059-0.352 microg for genistin, within 0.435-2.610 microg for rutin, within 0.020-0.121 microg for quercetin and within 0.053-0.319 microg for genistein. The correlation coefficient of each component is 0.999 6, 0.998 2, 0.998 9 and 0.999 9 respectively. The average recoveries of the four components are from 98.7% to 100.2%. The RSD of each group are 1.21%, 1.36%, 0.47% and 1.54% (n = 5).
CONCLUSIONThe method was accurate, repeatable and suitable to determine four effective components in Huaijiao pill. It can be use for quality control of Huaijiao pill.
Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Genistein ; analysis ; Isoflavones ; analysis ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Reproducibility of Results ; Rutin ; analysis ; Sophora ; chemistry
5.Role of PPARγ signaling pathway in osteogenic differentiation of rat bone marrow- derived mesenchymal stem cells in simulated microgravity.
Yiwen HUANG ; Rui YANG ; Si CHEN ; Jia SUN ; Rongping CHEN ; Zhen HUANG
Journal of Southern Medical University 2013;33(4):573-577
OBJECTIVETo explore the role of peroxisome proliferator-activated receptor γ (PPARγ) signaling pathway in osteo- blast differentiation of rat bone marrow mesenchymal stem cells (BMSCs) cultured in simulated microgravity.
METHODSRat BMSCs were cultured in simulated microgravity (by rotating clinostat) in the presence of 10 µmol/L pioglitazone, 10 µmol/L GW9662, or both pioglitazone and GW9662, with the cells cultured in normal gravity as the control group. After osteogenic induction for 14 days, the cells were stained with alizarin red for the bone nodules and with oil red-O for the fat cells, and the fat rate was calculated. ALP activity in the cells was determined in each group, and RT-PCR was performed to detect cellular expressions of PPARγ mRNA.
RESULTSPioglitazone significantly inhibited osteoblast differentiation of the BMSCs, whereas GW9662 promoted the cell differentiation by suppressing the activation of PPARγ.
CONCLUSIONWe hypothesize that the activation of PPARγ signaling pathway is one of the main mechanisms for inhibited osteoblast differentiation of rat BMSCs in simulated microgravity, and inhibiting PPARγ pathway activation can effectively prevent and treat microgravity-induced osteoporosis.
Animals ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Osteogenesis ; PPAR gamma ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Weightlessness Simulation
6.Screening of the target genes transactivated by PS1TP1 protein with suppression subtractive hybridization technique
Hui-Huang HUANG ; Dong JI ; Si-Yu WANG ; Wan-Zhen XU ; Zhi-Yuan JIA ; Ke LI
Chinese Journal of Experimental and Clinical Virology 2011;25(3):179-181
Objective To construct a subtractive cDNA library of genes transactivated by PS1TP1 protein with suppression subtractive hybridization ( SSH) technique. Methods Suppression subtractive hybridization technique and bioinformatics technique were used, the mRNA from HepG2 cells transfected with pcDNA3. 1(-)-PS1TP1 and pcDNA3. 1 (-) empty vector was isolated, respectively; cDNA underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Results The subtractive library of genes transactivated by PS1TP1 was constructed successfully. Sequence analysis was performed in 43 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 12 coding sequences were gotten, which consisted of 10 known and 2 unknown ones. Conclusion The obtained sequences may be target genes transactivated by PS1TP1 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity and cell apoptosis. This finding brought some clues for studying the biological functions of PS1TP1.
7.Epidemiological analysis of primary liver cancer in the early 21st century in Guangxi province of China.
Chun-Yan ZHANG ; Tian-Ren HUANG ; Jia-Hua YU ; Zhen-Quan ZHANG ; Ji-Lin LI ; Wei DENG ; Si-Yuan YE ; De-Nan ZHOU ; Zhen-Fang HE
Chinese Journal of Cancer 2010;29(5):545-550
BACKGROUND AND OBJECTIVEIn Guangxi province, from 1970s to 1990s, the mortality of primary liver cancer (PLC) ranked the first among a variety of malignant tumors. Investigating the epidemiological characteristics of PLC is very important for developing reasonable and effective treatment strategy, allocating health resources rationally, and evaluating the quality of PLC prevention and control. This study was to analyze the mortality and epidemiological characteristics of PLC in Guangxi province between 2004 and 2005.
METHODSMulti stage stratified cluster random sampling method was used to select 9 counties (cities or urban areas) as sample points. The residents' death causes between 2004 and 2005 were analyzed, and the epidemiological characteristics of PLC were investigated.
RESULTSIn the period of 2004-2005, the crude mortality of PLC was 34.39/100,000 in Guangxi province population (55.30/100,000 in men and 13.21/100,000 in women). The national population standardized mortality in 1964 was 22.17/100,000. The man to woman ratio of mortality was 4.19:1. PLC ranked as the first death cause among a variety of malignant tumors, and PLC related death accounted for 30.70% of all tumor related death cases. Age specific mortality of PLC was increased with age, rising significantly from 30 year old (from 25 year old in men and from 40 year old in women), and reached a peak at 75 year old.
CONCLUSIONSThe mortality of PLC shows a decreasing trend in Guangxi province in the early 21st century, and the starting age of PLC death peak postpones about 10 years than that in 1990s. It shows that the comprehensive prevention and control measures of PLC implemented in Guangxi province are fruitful. However, the PLC mortality in Guangxi province is still significantly higher than the national average level, and it still ranks as the first death cause in a variety of malignant tumors in Guangxi province. PLC mainly occurs in middle aged and elderly people. The prevention and treatment research of PLC still has a long way to go.
Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Liver Neoplasms ; epidemiology ; mortality ; Male ; Middle Aged ; Mortality ; trends ; Sex Distribution ; Young Adult
8.Development of microfluidic chip with adjustable concentration and pressure gradient for 3D cell culture
Si yuan LU ; Shao xi CAI ; Xiao zhen DAI ; Si jia CHEN ; Zhen SONG
Journal of Medical Biomechanics 2011;26(4):E335-E340
Objective To develop a microfluidic device with the adjustable concentration and pressure gradient for 3D cell culture in hydrogel and set up an in vitro model with the capability to closely simulate in vivo microenvironment for cell growth. Methods The microfluidic chip, with a middle channel for 3D cell culture and two side channels for delivering cell culture medium, was designed and fabricated using standard soft lithography and replica molding techniques. Its capability to generate concentration gradient, interstitial flow and image cell in situ was demonstrated. Results A simple microfluidic chip for 3D cell culture in hydrogel with the capability to generate the concentration and pressure gradient was obtained. At a flow rate of 2 μL•min-1 in each side channel, the concentration gradients remained constant after 3 h. The interstitial flow across the gel scaffold was generated by a 100 Pa pressure difference between two-side channels with the pressure gradient of 0.11 Pa/μm. Human adult dermal microvascular endothelial cells (HMVEC) were maintained in 3D culture with collagen type I and observed with confocal microscopy. Conclusions The microfluidic chip is simple and easy to operate and it can simulate the complicated microenvironment in vivo. The chip also allows the multiparameter control of microenvironment, facilitating the better understanding of interaction between cells and microenvironment.
9.Clinical study on combination of acupuncture, cupping and medicine for treatment of fibromyalgia syndrome.
Chang-du LI ; Xiao-ying FU ; Zhen-ya JIANG ; Xu-guang YANG ; Si-qin HUANG ; Qiong-fen WANG ; Jia LIU ; Yuan CHEN
Chinese Acupuncture & Moxibustion 2006;26(1):8-10
OBJECTIVETo observe the therapeutic effect of acupuncture at five mental points and moving cupping on the Hechelu of the back on fibromyalgia syndrome (FS).
METHODSSixty-six cases who conformed to the criteria were randomly divided into the treatment group treated with acupuncture at five mental points, moving cupping on the Hechelu of the back and amitriptyline, and the control group treated with amitriptyline. Clinical therapeutic effects were assessed with McGill Pain Questionnaire (MPQ) and HAMD depression scale.
RESULTSThe therapeutic effect of the treatment group was better than that of the control group with a significant difference between the two groups (P < 0.01).
CONCLUSIONCombination of acupuncture with cupping therapy is an effective therapy for fibromyalgia syndrome.
Acupuncture Points ; Acupuncture Therapy ; Biomedical Research ; Fibromyalgia ; Humans ; Pain Measurement
10.Clinical effects of percutaneous elastic intramedullary nail assisted by arthrography for the treatment of radial neck fractures in children
Hui-Min ZHOU ; Yi-Wen XU ; Chun-Jie TAO ; Jiang-Rong FAN ; Jing-Yang YOU ; Jia-Cheng RUAN ; Si-Qi SHEN ; Zhen WANG ; Yong ZHENG
China Journal of Orthopaedics and Traumatology 2024;37(9):899-904
Objective To explore clinical effect of closed reduction percutaneous elastic intramedullary nail assisted by arthrography in the treatment of radial neck fracture in children.Methods A retrospective analysis was performed on 23 chil-dren with radial neck fracture treated with arthrography assisted closed reduction and percutaneous elastic intramedullary nail internal fixation(arthrography with elastic nail group)from January 2019 to December 2022,including 12 males and 11 fe-males,aged from 2 to 12 years old with an average of(7.36±1.89)years old;According to Judet fracture types,14 children were type Ⅲ and 9 children were type Ⅳ.In addition,23 children with radial neck fracture were selected from January 2015 to December 2018 who were treated with closed reduction and percutaneous elastic intramedullary nail fixation(elastic nail group),including 11 males and 12 females,aged from 2 to 14 years old with an average of(7.50±1.91)years old;Judet classi-fication included 15 children were type Ⅲ and 8 children were type Ⅳ.Operative time and intraoperative fluoroscopy times were compared between two groups.Metaizeau evaluation criteria was used to evaluate fracture reduction,and Tibone-Stoltz evaluation criteria was used to evaluate functional recovery of elbow between two groups.Results Both groups were followed up for 12 to 24 months with an average of(16.56±6.34)months.Operative time and intraoperative fluoroscopy times of elastic nail group were(56.64±19.27)min and(21.13±7.87)times,while those of joint angiography with elastic nail group were(40.33±1 1.50)min and(12.10±3.52)times;there were difference between two groups(P<0.05).According to Metaizeau evaluation,11 patients got excellent result,9 good and 3 fair in joint angiography with elastic nail group,while in elastic nail group,5 ex-cellent,13 good,4 acceptable,and 1 poor;the difference between two groups was statistically significant(P<0.05).According to Tibone-Stoltz criteria,14 patients got excellent result,8 good,and 1 fair in joint arthrography with elastic nail group;while in elastic nail group,12 patients got excellent result,9 good,1 fair and 1 poor;there was no significant difference between two groups(P>0.05).Conclusion Compared to percutaneous elastic intramedullary nail fixation,closed reduction assisted by arthrography has advantages of reduced operation time,decreased intraoperative fluoroscopy frequency,and improved fracture reduction.Arthrography enables clear visualization of the anatomical structures of radius,head,neck,bone,and cartilage in children,facilitating comprehensive display of fracture reduction and brachioradial joint alignment.This technique more pre-cisely guides the depth of elastic intramedullary nail implantation in radius neck,thereby enhancing surgical efficiency and success rate.