3.A modified method for primary culture of human glomerular mesangial cells
Yanzhi YIN ; Limei ZHANG ; Jianwei JU ; Hongmei WANG ; Sha WANG
Chinese Journal of Tissue Engineering Research 2010;14(11):1939-1942
BACKGROUND:Primary culture of glomerular mesangial cells was less achievement ratio,short survival time,and less passage times.In particular,extraction of renal glomerulus remains difficult for culturing highly pure mesangial cell OBJECTIVE:To establish a more simple.high successful rate and good reproducibility method of human mesangial cells in primary cultureMETHODS:Kidneys jsolated from induction of labor with water bag voluntary were cut into pieces.and human mesangial cells were cultured with eugenic selection methods.Morphology was observed using inverted phase contrast microscope and transmission electron microscope,cell phenotype was detected using immunohistochemical method,and vimentin expression was observed using laser scanning confocal microscope.RESULTS AND CONCLUSION:The cultured mesangial cells were fusiform-shaped,irregular star-shaped,and slender.Organelle was rich in cytoplasm,cell process was clear,and microvillus was observed on the cell membrane.The cells expressed a-actin,myosin,vimentin,desmin but not expressed cytokeratin and Ⅷ factor.Laser scanning confocal microscope demonstrated that vimentin expression was positive and had the characteristics of fiber bundles.This suggested that the cultured intercapillary cells were coincidence with the characteristics of mesangial cell The renal corticaI tissue combined eugenic selection method was a simple and efficient method to culture human mesangial cells.
4.Effects of PDTC and curcumin on expression of tissue factor induced by anti-β2GPI in mice
Longfei XIA ; Hongmei JU ; Hong ZHOU ; Ting WANG
Chinese Journal of Immunology 2017;33(6):823-827
Objective:To investigate whether PDTC or curcumin had effect on anti-β2GPI-induced tissue factor (TF) expression in mice.Methods:BALB/c mice were pretreated with PDTC (100 mg/kg,once a day) by intraperitoneal injection (i.p.) or/and curcumin (50 mg/kg,once a day) by oral gavage for 3 consecutive days at 2 h before 500 μg of anti-β2GPI injections in subsequent experiments.Mouse peritoneal macrophages and aorta were collected,homogenized by sonication.The total RNA and protein were collected from each animal,TF expression was detected by Real-time quatitative PCR and TF activity kit.The phosphorylation of NF-κB p65 and c-Jun/AP-1 was determined by Western blot.Results:Anti-β2GPI cloud significantly upregulate TF expression and phosphorylation of NF-κB p65 and c-Jun/AP-1 in mouse peritoneal macrophages and aorta,compared with NR-IgG treated mice (P< 0.05).PDTC or/and curcumin could markedly attenuate anti-β2 GPI-induced TF expression,also inhibit activation of NF-κB p65 and cJun/AP-1 in the aorta and peritoneal macrophages respectively (P<0.05),but combination of two inhibitors had no synergistic effect.Conclusion:Both PDTC and curcumin could affect the expression of TF induced by anti-β2GPI in mice,indicatiug that PDTC and curcumin has the potential to prevent thrombosis in APS.
5.Effects of mTOR activation on anti-β2GPI/β2GPI-stimulated tissue factor expression in THP-1 cells
Longfei XIA ; Hongmei JU ; Hong ZHOU ; Ting WANG
Chinese Journal of Clinical Laboratory Science 2017;35(3):215-220
Objective To investigate the role of mammalian target of rapamycin(mTOR) in the expression of tissue factor(TF) from THP-1 cells induced by β2GPI/anti-β2GPIcomplex.Methods The THP-1 cells were treated with both β2GPI/anti-β2GPI and β2GPI/IgG-APS(β2GPI/IgG from APS patients) complexes.Rapamycin(100 nmol/L),the mTOR inhibitor,was used to exert the intervention experiment.The total RNA and proteins of the THP-1 cells were collected for detection.The mRNA expression level and activity of TF in THP-1 cells were detected by real-time quatitative PCR(RT-qPCR) and TF activity kit respectively.western blotwas used to determine the levels of mTOR and phosphorylated-mTOR(p-mTOR),and p38,p-p38,ERK1/2,p-ERK1/2,JNK,p-JNK,NF-κB p65 and p-NF-κB p65 in THP-1 cells were determined simultaneously.Results Both β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes chould significantly upregulate the mRNA expression and activity of TF,and the phosphorylation levels of mTOR in THP-1 cells(P < 0.05).Rapamycin markedly attenuated the mRNA expression and activity of TF and mTOR phosphorylation induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P < 0.05),and also inhibited the phosphorylation levels of p38,ERK1/2 and NF-κB p65 in THP-1 cells induced by β2GPI/anti-β2GPI and β2GPI/IgG-APS complexes (P < 0.05),but did not showed effects on the phosphorylation of c-Jun NH2-terminal protein kinase (JNK) (P > 0.05).Conclusion mTOR could be activated by β2GPI/antiβ2GPI complexes in THP-1 cells and play a crucial role for β2GPI/anti-β2GPI-induced TF expression in THP-1 cells.
6.Study on Loganin and Morronside Concentrations in Rat Serum Containing Liuwei Dihuang Formula by SPE-HPLC and the Variety of Different Combinations
Lingling JU ; Hongmei WEN ; Xin MA ; Huimin BIAN ;
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(03):-
Objective To establish a sensitive and specific liquid chromatography method for the determination of loganin and morroniside in rat serum containing Liuwei Dihuang Formula(LDF).Methods Loganin and morroniside were extracted from the rat serum using SPE column,then separated on a C_(18)column.A HPLC method for determining serum concentration of mor- roniside and loganin in rats before and after administration LDF was established.Results A good linear calibration curve of lo- ganin and morroniside was obtained.The recovery of the method was in the range of 95 %~105 %.The extracted recovery was more than 75 % ,and within-day and intra-day precisions were less than 12 %.Conclusion This method is sensitive, simple and rapid for monitoring of drug level in the study on pharmacokinetics and different combination of LDF.
7.Clinical Distribution Characteristics and Drug-resistance of AmpC-producing Enterobacter cloacae in ICU
Hongmei CHEN ; Qinfang TANG ; Hui ZHU ; Huixiang JU ; Mingzhong SUN
Chinese Journal of Nosocomiology 2009;0(18):-
OBJECTIVE To explore the clinical distribution characteristics and the drug-resistance of AmpC-producing Enterobacter cloacae in ICU. METHODS Seventy-eight strains of E. cloacae were isolated in ICU from Jan 2005 to Jan 2006 and antimicrocal sensitivities were determined by K-B method. RESULTS Among 78 E. cloacal strains derived from the lower respiratory tract and wound secretion,32 (41.0%) were AmpC producing and were sensitive to imipenem (96.9%) and merapenem (93.8%). CONCLUSIONS AmpC-producing E. cloacae is one of the main pathgens of nosocomial infection in ICU,imipenem and merapenem are the first choice to treat the infection.
9.Prediction of the long-term functional prognosis of patients with cerebral infarction by the classification of Oxfordshire community stroke project
Jin QIAO ; Hongmei CAO ; Xichi JU ; Feng GUO ; Qiumin QU ; Chengbin WU
Chinese Journal of Tissue Engineering Research 2005;9(9):182-183
BACKGROUND: Oxfordshire Community Stroke Project(OCSP) is a new type of clinical classification for subdividing cerebral infarction(CI) conducted on the basis of a large-scale of investigation of stroke in the population of Oxfordshire Community, England in 1991. This kind of classification completely bases on the clinical manifestations without the help of diagnostic instruments, which can predict site and size of the lesion and the involved vessels.OBJECTIVE: To acknowledge the clinical classification of CI patients with Bamford's OCSP and its significance in predicting their long-term functional prognosis.DESIGN: Clinical observation, comparison and verified study based on patients.SETTING: Neurological department in a university hospital.PARTICIPANTS: Between January 1st and December 31st 2001, totally 126 patients with cerebrovascular disease were hospitalized in the Neurological Department of First Affiliated Hospital of Xi' an Jiaotong University,including 82 males and 44 females.METHODS: Totally 126 in-patients with CI were subdivided with Bamford's OCSP classification, and their disablity was assessed with Barthel index (BI)and modified Rankin Scale(mRS) when they were discharged and 3 months and 6 months later.MAIN OUTCOME MEASURES: Classification of patients with CI and their scores of BI and mRS when they were discharged and 3 months and 6months later.RESULTS: Of the 126 patients, 8(6. 3% ) patients were confirmed of total anterior circulation infarction (TACI), 29 (23. 0% ) of part anterior circulation infarction(PACI), 78(61.9% ) of lacunar infarction(LACI), 11(8.7%) of posterior circulation infarction(POCI) . They were followed-up for 6 months, and meanwhile 12 patients died. Of the other 114 cases the prognosis of TACI was the worst, while the prognosis of POCI and LACI was relatively better than that of PACI.CONCLUSION: CI is predominated by LACI. OCSP is effective for predicting long-term functional prognosis of patients with CI. But it is necessary to make multi-center prospective study on a much larger scale of samples of disease.
10.Comparison of Mutant Prevention Concentrations of 3 Fluoroquinolones in 104 Escherichia coli Isolates
Yanping LUO ; Qinchun LI ; Liyan YE ; Zhongqiang YAN ; Xinxiu LIANG ; Leili WANG ; Hongmei JU
Chinese Journal of Nosocomiology 2009;0(17):-
0.05).Our results showed that,for recommended oral doses of ciprofloxacin and moxifloxacin,44.2% of E.coli isolates from the intestine of the health population,and 29.5%,18.7% and 10.7% of isolates from the three sterile sites of the patients,would be selected as resistant mutants.When ciprofloxacin and moxifloxacin were taken by injection route,the ratio of the selection of resistant mutants would be 9.3% for E.coli isolates from the intestine of the health population and 8.2%,6.2% and 8.9% for the three sterile sites of the patients,respectively.The maximum attainable concentration of levofloxacin in serum showed little distinction between the oral and the other routes.CONCLUSIONS The values of MIC and MPC of three fluoroquinolones in E.coli isolates from different populations and sites show no association.The values of MPC couldn't be predicted by the MIC.The values of MPC and MPC90 of three drugs show no significant discrepancy for tested isolates,these E.coli strains are isolated from the intestine of heath persons,and from the blood,ascites,bile of patients.