1.Effect of application of uridine diphosphate-glucose on self-repairment potentiality of immature cerebral white matter invivo
Fengxia MAO ; Huijin CHEN ; Longhua QIAN ; Wenjuan LI
Journal of Clinical Pediatrics 2013;(11):1059-1065
Objectives To explore the effect of application of uridine diphosphate-glucose (UDP-glucose) on self-repairment potentiality of immature white matter (WM) in vivo. Methods Five-day-old rats were randomly divided into sham, periventricular leukomalacia (PVL) and UDP-glucose groups. The PVL model was constructed in the PVL and UDP groups, and UDP-glucose (2000mg/kg) was induced by an intraperitoneal injection at once to the rats of UDP group. PVL in-duced proliferation and differentiation of WM-glial progenitor cells invivo were detected by using the three-label lfuorescent immunoanalysis, the apopotosis in WM cell was observed by TUNEL test, and the pathology of WM and myelination were evaluated by light and electron microscopy at day 7 and day 21 after PVL model construction. Results The numbers of new WM-progenitors (NG2+), oligodendrocytes (OLs) progenitor marker (O4+), OL precursors, cyclic nucleotide phosphodiesterase (CNPase+), immature OLs and myelin basic protein (MBP+), and mature OLs in the UDP-glucose group are signiifcantly grea-ter than those in the PVL group at each time interval after induction of PVL (P<0.05). The numbers of the apoptotic cells in UDP-glucose group are less than those in the PVL groups. Under light and electron microscopy, the white matter pathological changes and myelination were found to be better than those in the PVL group (P<0.05). Conclusion The application of UDP-glucose can induce the WM-progenitors to activate, proliferate and differentiate into immature and mature OLs. UDP-glucose can also signiifcantly reduce the apoptotic rate of the WM-new glia cells;improve the white matter pathological changes and the myelin formation.
2.Correlation between MMP12 expression in airway macrophages and pulmonary neurogenic substance P in asthmatic patients
Junyang XU ; Xinlan MAO ; Chuanen ZHOU ; Mengzhang HE ; Yan AI ; Jianhong XIE ; Fengxia JIANG ; Jiancheng Lü
Chinese Journal of Immunology 2016;(2):247-250
Objective:To explore the role of matrix metalloproteinase 12 (MMP12) in airway macrophages and pulmonary neu-rogenic substance P ( SP ) in the pathogenesis of asthma by analyzing their relationship in different categories of asthmatic patients.Methods:Twenty patients of asthma remission phase ( remission asthma group ) , twenty ones of mild acute exacerbation asthma (mild asthma group) and twenty healthy adults (normal control group) were included,respectively.After lung function was measured,the numbers of macrophage in induced sputum were counted.The expression levels of MMP12 mRNA and protein in sputum macrophages were detected by quantitative reverse transcription polymerase chain reaction and Western blot.The concentration of sputum SP was assayed by enzyme immunometric assay.Results: ( 1 ) Compared with the subjects in normal control group, forced expiratory volume in 1 second%predicted ( FEV1 ) and forced expiratory flow rates at 50% of the forced vital capacity % predicted ( FEF50 ) were much lower and the numbers of sputum macrophages were much higher in the patients in different asthmatic groups.Compared with the patients in remission asthma group,FEV1 and FEF50 were much lower in the ones in mild asthma group.(2) MMP12 expressions in the macrophages and the concentrations of SP in sputum were significantly increased in the patients in different asthmatic groups compared with those in normal control group;Furthermore,MMP12 and SP in mild asthma group were much higher than in remission asthma.(3) In all patients from different asthmatic groups,mRNA expressions of MMP12 in the macrophages were positively correlated with the levels of sputum SP or the numbers of sputum macrophages,whereas negative correlations between mRNA expressions of MMP 12 and FEV 1 or FEF50 were observed.Conclusion: The regulatory imbalance of macrophages′MMP12 and pulmonary neurogic SP may participate in the pathogenesis of asthma and become the potential targets for asthma therapy.
3.Culture of glial lineage progenitor cells derived from white matter and establishment of its oxygen-glucose deprivation models
Wen-Juan LI ; Hui-Jin CHEN ; Long-Hua QIAN ; MAO-FengXia
Chinese Journal of Neuromedicine 2013;12(11):1106-1111
Objective To in vitro culture glial lineage progenitor cells derived from periventricular white matter in neonatal rats,and establish oxygen-glucose deprivation models of progenitor cells to further explore the mechanism of neurogenesis in the ischemic white matter injury.Methods The progenitor cells were isolated and cultured from the white matter of 3-d-old neonatal rats.Cells were passaged every 3 or 4 d,and were further induced differentiation.The primary and passaged neurospheres and their differentiated cells were respectively identified by immunocytochemical stanning of NG2 (Neurospheres marker) and O4 (oligodendrocyte precursor marker).The third-or fourth-generation progenitor cells were used to establish the ischemic model of oxygen-glucose deprivation (OGD).The influence of different OGD periods (30,45 and 60 min,2 and 3 h) in the survival rates of progenitor cells was assessed 24 h after success of model making by both CCK-8 and Hoechst 33342/PI stainning.Results The progenitor cells obtained from the white matter had the capacity of forming neurospheres and reproduction which showed NG2 immunoreactive positivity,and could be induced differentiation into O4-positive oligodendrocyte precursors.No apoptotic or necrotic cells were observed in normal cultured NG2-positive progenitor cells.The apoptotic and necrotic cells tended to increase significantly with the extension of OGD period gradually.The survival rates of progenitor cells in turn were (85.94±3.06) % for 30 min OGD,(62.68±2.66) % for 45 min OGD,(45.09±2.24) % for 60 min OGD,(36.70±2.84) % for 2 h OGD and (22.01±3.00) % for 3 h OGD,respectively,with significant differences between each two groups (P<0.05).Conclusion The oxygen-glucose deprivation models of glial lineage progenitor cells derived from periventricular white matter of neonatal rats are successfully established,which may provide the foundation for further exploring the mechanism of neurogenesis in the ischemic white matter injury.
4.Calcium-sensing receptor overexpression induces proliferation and differentiation of immature white matter progenitor cells after ischemia in vitro
Fengxia MAO ; Mengyuan LEI ; Xinru CHENG ; Ju ZHANG ; Qianya XU ; Zanyang SHI ; Suge HAN ; Qian ZHANG
Chinese Journal of Neuromedicine 2021;20(4):325-330
Objective:To explore the promoted role of calcium-sensing receptor (CaSR) overexpression in proliferation and differentiation of immature white matter progenitor cells after ischemia in vitro. Methods:Periventricular white matter cells from 5-d-old rats were cultured in vitro and divided into control group, oxygen-glucose deprivation (OGD) group, OGD+gadolinium chloride (GdCl 3) group, and OGD+ CaSR silenced group. CaSR expression was agitated by GdCl 3, and CaSR gene expression was inhibited by gene silencing. CaSR mRNA levels 24, 48 and 72 h, and 7 and 14 d after OGD were detected by real-time fluorescence quantification-PCR (RT-qPCR); cell proliferation 48 h after OGD was detected by inverted microscope, and cell differentiation 24, 48, and 72 h, and 7 and 14 d was detected by double immunofluorescence staining. Results:(1) CaSR mRNA expressions: CaSR mRNA expressions in OGD group 48 and 72 h and 7 d after OGD were statistically higher than those in control group ( P<0.05). Forty-eight and 72 h, and 7 and 14 d after OGD, CaSR mRNA expressions in OGD+GDCL 3 group were statistically higher than those in control group and OGD group ( P<0.05); the CaSR mRNA expressions in the OGD+CaSR silenced group were significantly lower than those in the control group ( P<0.05). (2) Cell proliferation and differentiation: 48 h after OGD, the cell sphere diameter of OGD group ([75.26±26.07] μm) was significantly increased as compared with that of control group ([57.96±18.92] μm, P<0.05); the cell sphere diameter of OGD+GdCl 3 group ([91.92±21.82] μm) was significantly increased as compared with that of control group and OGD group ( P<0.05); and the cell sphere diameter of OGD+ CaSR silenced group ([24.09±8.34] μm) was significantly shorter than that of control group and OGD group ( P<0.05). At 48 and 72 h after OGD, the number of O4 +/CaSR + olidoendrocyte precursor cells (OPCs) in OGD group was significantly larger than that in control group, that in OGD+GdCl 3 group was significantly larger than that in control group and OGD group, and that in OGD+ CaSR silenced group was significantly smaller than that in control group and OGD group ( P<0.05). Conclusion:CaSR overexpression could promote the proliferation and differentiation of progenitor cells to OPCs.
5.Comparative field study on high flow rate samplers for respirable fraction-A solution to smaller collected masses.
Limin WANG ; Fengxia HU ; Zhenglun WANG ; Jiaojun LIANG ; Jichao LI ; Geshi MAO ; Wwili SONG ; Guilin YI ; Lei ZHAO ; Jiabing WU ; Michael KOOB ; Weihong CHEN ; Dirk DAHMANN ; Lei YANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2016;34(1):32-36
OBJECTIVEDust sample mass gain is too smaller to satisfy the limit of detection (LOD) even in most cases during dust sampling at workplaces nowdays, especially for respirable fraction. Therefore, it is aimed to solve the problem by increasing sample load with high flow rate samplers.
METHODSIn A and B two shipyards respirable welding fume was sampled by high flow rate cyclone samplers of FSP-10 (10 L/min) for 2-2.5 hours and normal flow rate FSP-2 (2 L/min) for 3-4 hours with a stratigy of parallele sampling at the same workpalce, in order to compare their mass gain, coincidence rate with LOD, and airborn dust concentration.
RESULTSSample mass gain of 0.97±0.40 mg and 1.61±0.86 mg respectively in the two factories by FSP-10 was significantly higher than that of 0.29±0.12 mg and 0.51±0.27 mg by FSP-2 (t-test, P<0.05 in both cases) , increasing herewith the coincidence rate with LOD from 26.8% (when sampling with FSP-2, calculated together with samples of the two factories) to 89.7%. However there was no significant difference in dust concentrations by the two different samplers, 0.53±1.88 vs 0.73±1.61 mg/m(3) by FSP-2 and FSP-10 in the shipyard A and 1.14±1.78 vs 1.01±1.63 mg/m(3) in the factory B (t-test, P>0.05 in every case) . In addtion, sample loading by FSP-2 was found to be correlated to sampling time (R(2)=0.7906, y=0.002 6x) , therefore, it has to sample for ≥192.3 min to meet the LOD (0.5 mg) in case of normal flow rate.
CONCLUSIONBy using of high flow rate cyclone FSP-10 the problem of LOD could be solved, along with increased sample mass and similar respirable dust concentration by the two samplers. Some techincal improvements of FSP-10 and increasing of LOD coincidence rate by other methods was also disscussed.
Air Pollutants, Occupational ; analysis ; Construction Industry ; Dust ; analysis ; Environmental Monitoring ; instrumentation ; Occupational Exposure ; Ships ; Workplace
6.Study on the relationship between perinatal hemodynamics and retinopathy of prematurity
Min SHEN ; Xinru CHENG ; Mengyuan LEI ; Zanyang SHI ; Junbo RONG ; Shuanfeng FANG ; Shuling XU ; Peige XIA ; Suge HAN ; Lili WANG ; Fengxia MAO ; Qianya XU ; Li WANG ; Qian ZHANG
Chinese Journal of Applied Clinical Pediatrics 2020;35(19):1485-1489
Objective:To explore the correlation between the index of hemodynamics in perinatal period and retinopathy of prematurity(ROP), so as to provide basis for the better prevention and treatment of ROP.Methods:From May 2017 to April 2019, the preterm infants were admitted to the Neonatal Intensive Care Unit of the First Affiliated Hospital of Zhengzhou University at birth and were hospitalized for more than 2 weeks, gestational age ≤ 35 weeks and birth weight ≤ 2 500 g. They were selected as the study objects.The perinatal data including heart rate, blood pressure, patent ductus arteriosus, ventricular septal defect, and NT-proBNP level on the 1 st, 7 th and 14 th day, respectively after birth were collected.They were divided into ROP group and non ROP group according to the results of the retinopathy screening report.The influencing factors of ROP were screened out by univariate analysis and multivariate regression analysis. Results:A total of 1 119 subjects were included, 105 infants with ROP were detected, and the prevalence of ROP was 9.4%.Among them, 12 cases of pre-threshold lesion type 1 and threshold lesions required treatment, accoun-ting for 1.07% of screened preterm infants .Univariate analysis and multivariate regression analysis revealed that gestational age, birth weight, total oxygen therapy time, and intrauterine growth restriction were all factors affecting ROP, and 2 hemodynamic related indicators, such as the level of NT-proBNP in plasma on the 14 th day after birth, and placenta previa or abruption were also related to ROP( OR=0.604, 0.647, 1.276, 2.361, 1.688 and 2.506, respectively, all P<0.05). Conclusion:The hemodynamic changes in perinatal period may be involved in the formation of ROP, and it is necessary to further clarify its mechanism.
7.Inhibitory effect of jianpi-jiedu prescription-contained serum on colorectal cancer SW48 cell proliferation by mTOR-P53-P21 signalling pathway.
Fengxia LIN ; Sanlin LEI ; Jin'an MA ; Li SHI ; Dan MAO ; Shaofan ZHANG ; Jianhua HUANG ; Xinyi LIU ; Dengfeng DING ; Yingjin ZHANG ; Sifang ZHANG
Journal of Central South University(Medical Sciences) 2016;41(11):1128-1136
To investigate the effect of jianpi-jiedu (JPJD) prescription-contained serum on colorectal cancer SW48 cell proliferation and the underlying mechanisms.
Methods: Crude extract from JPJD was made by water extract method and the main components of crude extract from JPJD were analyzed by ultra-performance liquid phase high resolution time of flight mass spectrometry (UPLC-Q-TOF/MS). The low, medium, and high-concentration of JPJD-contained serum were prepared by the serum pharmacological method. The effect of serum containing JPJD on SW48 cell proliferation was determined by MTT assay. The cell cycle was detected by flow cytometric method. The protein levels of mammalian target of rapamycin (mTOR), phospho-mTOR, P-P53, and -P21, and the mRNA level of mTOR were examined by Western blot and RT-PCR, respectively.
Results: Seven compounds including calycosin-7-glucoside, astragaloside, ginsenoside-Re, ginsenoside-Rb1, glycyrrhizinic acid, apigenin, atractylenolide-II were identified. MTT assays demonstrated that the SW48 cell proliferation was inhibited by medium and high concentration of JPJD-contained serum and the percentages of cells at G1 phase in SW48 cell cultured in the medium and high concentration of JPJD serum group were significantly higher than those in the control group (P<0.05). Meanwhile, the levels of mTOR mRNA and phospho-mTOR protein in the medium and high concentration of JPJD serum groups were substantially lower than those in the control group (P<0.05). Conversely, the expressions of phospho-P53 and P21 protein were significantly increased in the medium and high concentration of JPJD serum group compared with those in the control group.
Conclusion: JPJD prescription-contained serum can inhibit SW48 cell proliferation, which may be related to mTOR-P53-P21 signaling pathways.
Animals
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Apigenin
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Blotting, Western
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Cell Cycle
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Cell Division
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Cell Proliferation
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drug effects
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genetics
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Colorectal Neoplasms
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Cyclin-Dependent Kinase Inhibitor p21
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Flow Cytometry
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Ginsenosides
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Glycyrrhizic Acid
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Humans
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Lactones
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Phosphorylation
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genetics
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RNA, Messenger
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Saponins
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Sesquiterpenes
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Signal Transduction
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TOR Serine-Threonine Kinases
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drug effects
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Triterpenes
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Tumor Suppressor Protein p53
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drug effects