2.Establishment of MDCK cell models expressing human MATE1 or co-expressing with human OCT1 or OCT2.
Hong-mei LEI ; Si-yuan SUN ; Li-ping LI ; Mei-juan TU ; Hui ZHOU ; Su ZENG ; Hui-di JIANG
Acta Pharmaceutica Sinica 2015;50(7):842-847
To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals
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Biological Transport
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Cimetidine
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pharmacology
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DNA, Complementary
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Dogs
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Drug Interactions
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Humans
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Madin Darby Canine Kidney Cells
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Metformin
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pharmacology
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Organic Cation Transport Proteins
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genetics
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metabolism
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Transfection
3.The development and implementation of hospital-community Interactive nursing mode for patients with diabetes
Mei LI ; Ruizhen XU ; Tao SUN ; Suxia CHEN ; Shaoru SU ; Junying ZHAI ; Zhenzhen ZHANG
Chinese Journal of Nursing 2009;44(8):703-704
This paper introduces the development and implementation of hospital-community interactive nursing mode for patients with diabetes. The interactive mode between community and hospital,specialist nurses and community nurses,as well as patients and nurses was formed to provide timely,convenient,continuous and whole-process nursing care for diabetic patients through the setting up and services of hespital-based diabetes care team,two-way transfer center between hospital and commu-nity,hospital-based education and community-based visit team,community-based self-management group as well as green chan-nel of priority treatment,inspection and hospitalization for diabetic patients. The implementation of the interactive mode achieved good effect and was approved by the patients. The patient satisfaction rate was 96.5%.
4.Retrospective study of atrial fibrillation patients diagnosed by coronary artery angiography
Cheng-Qian YIN ; Zhi-Zhong LI ; Jing-Mei ZHANG ; Tong LIU ; Su WANG ; Tao SUN ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To identify the different causes of atrial fibrillation (AF) in Chinese by coronary artery angiography,and to compare the episode of stroke in different patients.Method A total of 782 adult patients diagnosed with AF were identified from Anzhen Hospital over a 8-year period.There were 273 patients with rheumatic valvular AF and 509 patients with nonvalvular AF.The results of electrocardiogram,echocardiography and coronary artery angiography were retrospectiwely analyzed to find out the proportion of causes and to compare the episode of stroke in patients with rheumatic valvular AF with those having nonvalvular AF.Results The patients with rheumatic valvular atrial were significantly younger than of coronary heart disease (52.40?5.03 years old vs 64.30?3.25 years old,P
5.Gastric cancer stem cells in tumor invasion and metastasis and its influence on angiogenesis ability
Lixian ZHANG ; Ning ZHANG ; Shuangzhen YUAN ; Yumei CHEN ; Xinai SU ; Jianshun SUN ; Lihua WANG ; Mei LIN
Chinese Journal of Tissue Engineering Research 2016;20(32):4738-4744
BACKGROUND:Gastric cancer stem cels involved in the neoadjuvant chemotherapy and conventional treatment are closely associated with relapse of gastric cancer. However, this conclusion has not yet been confirmed.
OBJECTIVE:To investigate the effects of gastric cancer stem cels in tumor invasion and metastasis and its effect on angiogenesis ability.
METHODS:We prepared nude mouse models of gastric cancer to isolate and culture gastric cancer stem cels. Harvested gastric cancer stem cels were detected in cel scratch test, ring test, inhibition rate test, cel migration test and tumorigenicity test.
RESULTS AND CONCLUSION:After 7 days of culture, the cels exhibited adherent growth but a lack of regularity that most cels were in a tadpole shape. In the cel scratch test, the scratch width was significantly different at 0 and 24 hours (P < 0.05). Under an inverted microscope, the cels were found to form a ring in the ring test. The 50% inhibiting concentration of gastric cancer stem cels induced by oxaliplatin was significantly lower than that induced by 5-fluorouracil (P < 0.05). The number of cels passing through the basilar membrane was significantly increased after cel migration (P < 0.05). After implantation of gastric cancer stem cels, the gastric tissue quality was significantly higher than that in normal nude mice of gastric cancer (P < 0.05). These findings indicate that gastric cancer stem cels responsible for tumor invasion and migration have stronger angiogenesis ability.
6.Pharmacokinetics of escitalopram in Chinese healthy volunteers after single and multiple oral administration
Tian-Mei SI ; Yi LIU ; Zhen-Guo ZHAO ; Li-Li SUN ; Yun-Ai SU ; Chun-Mei GUO ; Hong-Yan ZHANG ; Liang SHU
The Chinese Journal of Clinical Pharmacology 2009;25(4):308-311
Objective To explore the pharmacokinetics of single and multiple oral 20 mg escitalopram in healthy Chinese volunteers. Methods A total of 12 subjects participated in the study. Escitalopram 20 mg was given orally once on day 1 and days 8 to 14 in the fast condition. Sequen-tial blood samples were collected over 144 hours on day 1 and 14 and a predose sample was obtained on day 12 to 14. Escitalopram concentrations in plasma were determined by a validated HPLC fluorescence method. The pharmacokinetic parameters were calculated with DAS software. Results Escitalopram disposition on oral administration is characterized by a two-compartment pharmacokinetic model. The mean t1/2 is 41.09 h, Cavis (76.4±26.8) μg·L-1, AUCss is (1832.4±642.4) μg·h ·L-1,AUC0-t, and AUC0-∞are (4765.9±2171.0) and (5385.6±2851.2) μg ·h·L-1, respectively; tmaxis (3.2±1.3) h;t1/2 is (41.1±17.7) h;CLis 5.0 L·h1. The mean accumulation index of AUC ( RAUC) is ( 1.2 ±0.3 ). Conclusion Escitalopram pharmacokinetics in healthy Chinese subjects given 20 mg once daily dosing regimen were characterized by a two-compartment pharmacokinetic model. The state -concentration oc-curre after 7 days of continuously dosing. There is no accumulation after continuously dosing.
7.Cost-effectiveness evaluation of seven screening strategies for nasopharyngeal carcinoma.
Dong-ping RAO ; Qing LIU ; Su-mei CAO
Chinese Journal of Oncology 2012;34(7):549-553
OBJECTIVETo evaluate the cost-effectiveness of different screening strategies for nasopharyngeal carcinoma (NPC) and recommend a preferable NPC screening strategy.
METHODSA Markov simulation model was constructed based on the natural history of NPC. Seven strategies (A. Annual screening; B. Annual screening for (Epstein-Barr virus, EBV) EBV-seropositive subjects, triennial screening for seronegative subjects; C. Biennial screening; D. Triennial screening; E. 4-year screening; F. 5-year screening; G. 6-year screening) were evaluated. The NPC-pickup rate, cost, quality-adjusted life-years (QALYs) and incremental cost-effectiveness ratio (ICER) were calculated.
RESULTSThe ICERs of the 7 strategies were 83 111.6, 47 768.9, 50 164.7, 40 016.2, 34 272.8, 32 215.6, and 32 248.0 Yuan/QALY, respectively. The discounted QALYs of the strategies were 23 079.9, 22 955.6, 22 810.4, 22 636.5, 22 522.7, 22 445.0, and 22 361.9 years, respectively. The ICERs of the strategies were less than three times of the average per capita gross domestic product (89 976 Yuan) in China in 2010. The strategy A achieved a highest NPC pick-up rate (81.7%), a highest discounted QALY and a smallest number of NPC death (681), but a highest discounted cost and a greatest ICER. Compared with the strategy A, the strategy B achieved a little smaller NPC pick-up rate (73.1%), a little smaller number of NPC death (707), however, the ICER of the strategy B decreased by 38.2%.
CONCLUSIONThe strategy B (annual screening for EB virus seropositive subjects and triennial screening for seronegative subjects) is a preferable option for NPC screening.
Adult ; Carcinoma ; China ; epidemiology ; Cost-Benefit Analysis ; Early Detection of Cancer ; economics ; methods ; Epstein-Barr Virus Infections ; diagnosis ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Markov Chains ; Mass Screening ; economics ; methods ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; mortality ; Quality-Adjusted Life Years ; Survival Rate
8.Analysis of Epstein-Barr virus with BamHI "f" variant and XhoI-loss of LMP1 gene in nasopharyngeal carcinoma.
An-jia HAN ; Yong-sheng ZONG ; Min ZHANG ; Su-mei CAO ; Su-xia LIN ; Ying-jie LIANG
Chinese Journal of Pathology 2003;32(6):534-538
OBJECTIVETo investigate the genomic variation of Epstein-Barr virus (EBV) and its significance in nasopharyngeal carcinogenesis.
METHODSForty nasopharyngeal carcinoma (NPC) biopsy tissues were used for detection of EBV BamHI f variant and LMP1 XhoI-loss by polymerase chain reaction (PCR), nested PCR, and RFLP (restriction fragment length polymorphism). Forty-eight samples of peripheral blood mononuclear cells (PBMC) taken from apparently healthy adult individuals were used for detection of LMP1 XhoI-loss. Three samples of amplified LMP1 exon 1 DNA from B95-8 cell line and 2 NPC tissues (one having XhoI-loss and the other having Wt-XhoI/XhoI-loss) were sequenced.
RESULTSThirty out of the 40 NPC cases (30/40, 75%) harbored EBV BamHI f variant and the remaining 10 (10/40, 25%) harbored BamHI F prototype. Thirty out of the 39 NPCs (30/39, 76.9%) showed single EBV LMP1 XhoI-loss, 7 (7/39, 18.0%) showed single LMP1 Wt-XhoI (presence of a XhoI site in exon 1 of LMP1 gene, as in B95-8 cell line), and 2 (2/39, 5.1%) showed both LMP1 Wt-XhoI and XhoI-loss. Thirty-eight of the 39 NPCs (97.4%) showed EBV LMP1 XhoI-loss or/and BamHI F variant. In the NPC tissue (1 case only) showing the prototype of Wt-XhoI/BamHI "f", there were several base substitutions, including 5 missense mutations and 2 silent mutations present in LMP1 exon 3, on DNA sequencing. On the other hand, 10 out of the 48 samples of PBMC taken from apparently healthy individuals could be amplified successfully by nested PCR for detection of LMP1 XhoI site. All of these 10 samples carried the prototype of EBV LMP1 Wt-XhoI.
CONCLUSIONSThe majority of EBV present in neoplastic cells of NPC is of BamHI "f" variant and/or possesses LMP1 XhoI-loss, as compared with that in healthy individuals. This genomic variation of EBV may bear some roles in the development and progression of NPC.
Adult ; Aged ; Binding Sites ; genetics ; DNA, Viral ; genetics ; metabolism ; Deoxyribonuclease BamHI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Mutation ; Nasopharyngeal Neoplasms ; virology ; Sequence Deletion ; Viral Matrix Proteins ; genetics
9.Determine resource chemical component in ginkgo pollen simultaneously by UPLC-TQ-MS.
Cheng-mei XU ; Hao REN ; Da-wei QIAN ; Guang-tian SUN ; Shu-lan SU ; Sheng GUO ; Zhen OUYANG ; Jin-ao DUAN
China Journal of Chinese Materia Medica 2015;40(11):2157-2162
The present study is to determine the flavonoid glycosides, terpene lactones, biflavones, gingko acid and procyanidins of ginkgo pollen. UPLC-TQ-MS technology was used for the determination of 24 kinds of resource chemical composition in ginkgo pollen qualitatively and quantitatively. The results shows that the contents of rutin, quercetion 3-O-[4-O-(α-L-rhamnosyl )-β-D-glucoside] and kaempferolis were 120.9, 114.0, 222.1 μg x g(-1). In this paper, the contents of 24 kinds of chemical components of ginkgo pollen were determinated by UPLC-TQ-MS for the first time. This method is simple and quick, which will be benefit for recycling utilization of ginkgo pollen.
Chromatography, High Pressure Liquid
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methods
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Flavonoids
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analysis
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Ginkgo biloba
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chemistry
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Mass Spectrometry
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Pollen
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chemistry
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Proanthocyanidins
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analysis
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Rutin
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analysis
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Terpenes
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analysis
10.Effect of host derived matrix metalloproteinase on the degradation of root dentin collagen.
Dong-mei YANG ; Yu-jing LI ; Ying SU ; Yi-lin SUN
Chinese Journal of Stomatology 2006;41(5):275-278
OBJECTIVETo evaluate the effect of dentin matrix metalloproteinase (MMP) on the degradation of root dentin collagen.
METHODSRoot dentin powder was demineralized with acetic acid (pH 4.0) at 4 degrees C for 14 d, then dialysed and centrifuged. Precipitation was divided into 7 groups, with 6 samples in each group, and each sample was 50.0 mg. One milliliter artificial saliva with a different reagent was added in each sample respectively. The reagents were 2 mmol/L APMA (MMP activator), 2 mmol/L EDTA, 100 mmol/L EDTA, 200 mmol/L EDTA, 0.2% and 0.02% chlorhexidine (MMP inhibitor), and the blank artificial saliva was taken as control. The amount of degraded collagen of each sample was determined with hydroxyproline assay kit. Scanning electron microscope was employed to observe the morphological and structural changes of root dentin which was demineralized or put into artificial saliva after being demineralized.
RESULTSThe mean amount of degraded collagen in APMA group was significantly higher than that in the blank group (P < 0.05). The mean amount of degraded collagen in 2 mmol/L, 100 mmol/L, 200 mmol/L EDTA, 0.02% and 0.2% chlorhexidine groups was dramatically lower than that of the APMA group and the blank (P < 0.01). SEM observation indicated that the structural integrity of the collagen network on root surface dentin still existed in root dentin surface after being demineralized alone, while collagenous fibril was destructed and the structural integrity on root dentin surface disappeared after being demineralized and treated by artificial saliva.
CONCLUSIONSMMP in root dentin can degrade root dentin collagen after being activated at low pH followed by neutralization. The results suggest that host MMP may play an important role in the process of dentin caries formation.
Collagen ; metabolism ; Dental Caries ; enzymology ; Dentin ; metabolism ; Humans ; In Vitro Techniques ; Matrix Metalloproteinases ; metabolism ; Tooth Root ; metabolism