1.The effect of overexpression HMGA2 on proliferation and cell cycle in human growth hormone-secreting pituitary tumor cells
Bingchao YAN ; Hongfu CHEN ; Jianyong JI ; Hui ZHANG ; Pengjin MEI
Chongqing Medicine 2013;(36):4371-4373
Objective To explore the effects of over-expression high-mobility-group A2(HMGA2) on proliferation and cell cycle in primary cultured human growth hormone-secreting pituitary tumor cells .Methods The protein expression of HMGA2 in growth hormone-secreting pituitary tumor cells transfected by HMGA2 overexpression plasmid (transfection group) were detected by Western blot .The effects of the HMGA2 overexpression on cell proliferation ,cell cycle were measured by (cell counting kit-8 , CCK-8) assay and flow cytometry respectively .Results HMGA2 protein of transfection group was higher than the blank load transfection group(tranfectied with pcDNA3 .1 plasmid) after 24 h (P<0 .05) .To compare with the blank load transfection group , cells overexpression of HMGA2 could drastically increase the ability of proliferation at 24 ,48 ,72 ,96 h and increase the S phase rati-o of cell cycle and decrease the G1 phase ratio of cell cycle ,the difference was statistically significant (P<0 .05) .Conclusion Over-expression of HMGA2 increases pituitary tumor cells proliferation ability and promote cells G 1 phase progress .
2.Antagonism of Bacillus spp.B_1 and B_2 Strains Against Fusarium oxysporum f.sp. pisi
Xiao-Mei LIU ; Xiou-Rong CHEN ; Jin-Ji PU ;
Microbiology 1992;0(03):-
The antagonism of two strains B1 and B2 of Bacillus spp. against pea root rot pathogen Fusarium oxysporum f. sp. pisi was studied. The result of pairing culture showed that B1 and B2 strains of Bacillus spp. had strong antifungal activity to the pathogen. The colonial color of the pathogen changed from gray to white, aerial hyphas increased and entangled into group after treatment with the cell-free fermentational filtrate of B1 or B2. Observation under optical microscope showed that hyphas and spores of the pathogen swelled and distorted with concentrated cytoplasm after treatment, the spores could not germinate or germinated abnormally . A lot of vesicles appeared at the top of the hyphas, and the hyphas stopped growing and broke finally, their cytoplasm spilled from the cell. The cell-free fermentational filtrate of B1 or B2 strains contained 1795.53?g/mL and 1345.93?g/mL protein respectively, from which two antifungal proteins of 103.5 kD (B1 ) and 127.6 kD (B2) were purified.
3.Effect of UVA irradiation on proliferation and NO/iNOS system of human skin fibroblast
Mingliang CHEN ; Guiying ZHANG ; Mei YI ; Xiao CHEN ; Ji LI ; Hongfu XIE ; Xiang CHEN
Journal of Central South University(Medical Sciences) 2009;34(8):705-711
dosage ( P<0.01).Conclusion UVA can inhibit the proliferation activity of human skin fibroblasts. It might be related to the up-regulation of iNOS gene expression and the over-secretion of NO induced by UVA.
4.Effect of microRNA-21-5p on hyperoxic acute lung injury in rats
Guoyue LIU ; Miao CHEN ; Hui JI ; Song QIN ; Hong MEI ; Tao CHEN ; Huajun CHEN
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;(1):23-27
Objective To investigate the effects of microRNA-21-5p (miR-21-5p) on hyperoxic acute lung injury (HALI) in rats and provide a theoretical basis for HALI gene therapy. Methods One hundred and sixty Sprague-Dawley (SD) rats were randomly divided into four groups with number table:hyperoxia control group, phosphate buffer saline (PBS) group, blank virus group and miRNA-21-5p group (each, n = 40). The rats in hyperoxia control group were fed directly in the hyperoxia box (oxygen concentration > 90%); in the other three groups, 200 μL PBS, 200μL slow virus and 200μL miRNA-21-5p slow virus were dropped into the nose respectively, and then they were fed in the hyperoxia box. The rats were exposed to hyperoxia in the boxes for 0, 24, 48 and 72 hours in all the groups, and at each time point, 10 rats were taken randomly from each group to perform arterial blood-gas analysis, calculate oxygenation index (OI) and respiratory index (RI). Afterwards the rats were sacrificed by blood-letting from carotid artery under intra-peritoneal anesthesia, and the lung tissues were obtained to measure the left lung wet/dry weight (W/D) ratio, hemotoxylin-eosin (HE) staining was made and the pathological changes of the right lung were observed under light microscope and the pathological score was measured. Results At 0 hour, the OI, RI, lung W/D ratio and the lung tissue pathology score in rats with hyperoxic injury had no statistically significant differences among the four groups (all P>0.05). With the extension of time, the level of OI was gradually reduced, and the levels of RI, pathologic score and W/D ratio of lung tissues were gradually increased. Compared with the hyperoxia control group, in miRNA-21-5p group, the levels of OI were increased significantly at 24, 48 and 72 hours after the exposure to hyperoxia [mmHg (1 mmHg = 0.133 kPa): 24 hours 358.10±29.25 vs. 306.19±37.23, 48 hours 336.67±29.27 vs. 269.70±29.00, 72 hours 323.81±19.05 vs. 203.81±43.40, all P < 0.05], whereas the levels of RI were decreased significantly (24 hours 0.23±0.05 vs. 0.31±0.06, 48 hours 0.28±0.07 vs. 0.38±0.06, 72 hours 0.30±0.04 vs. 0.46±0.07, all P <0.05), the pathologic scores were decreased significantly (24 hours 0.60±0.52 vs. 0.90±0.74, 48 hours 1.30±0.95 vs.2.90±1.20, 72 hours 1.90±0.88 vs. 4.70±1.57, all P < 0.05) and the levels of W/D ratio were decreased obviously (24 hours 3.77±0.38 vs. 4.14±0.46, 48 hours 3.83±0.31 vs. 4.56±0.34, 72 hours 3.89±0.31 vs. 5.32±0.27, all P<0.05). Compared with the hyperoxia control group, the index results of the PBS group and the blank virus group after staying in the box had no statistical significant differences at each time point (all P>0.05). Under the optical microscope, along with the prolongation of exposure to hyperoxia, the structure of alveoli was gradually disturbed, their walls fractured and damaged, alveolar septa widened, edematous, infiltrated with inflammatory cells and in part of the rats a small amount of red blood cell exudates could be seen, but the degree of lung pathological injury in miRNA-21-5p group was much milder than that of the other groups. Conclusion The rat persistently exposed to hyperoxia for 24 hours can establish the rat model of HALI successfully, and the miRNA-21-5p can protect the lung tissue from the damage to some degrees in HALI rats.
5.The molecular mechanism of antiapoptosis of typeⅡ alveolar epithelial cell by microRNA-21-5p
Song QIN ; Miao CHEN ; Hui JI ; Guoyue LIU ; Tao CHEN ; Kang LI ; Hong MEI
Chinese Critical Care Medicine 2015;(7):574-578
ObjectiveTo study the effect of hydrogen peroxide (H2O2) in inducing apoptosis of typeⅡalveolar epithelial cell (AECⅡ) after overexpression by adenoviral transfection of micro RNA-21-5p (miR-21-5p), and to explore the mechanism of its anti-apoptosis.Methods Subculture AECⅡ were randomly divided into four groups: normal control group (normal saline), H2O2 challenge group ( 0.5 mmol/L H2O2), miR-21-5p overexpression group (miR-21-5p adenovirus+ 0.5 mmol/L H2O2), miR-21-5p negative transfection group (adenovirus void+0.5 mmol/L H2O2). Transmission electron microscopy and flow cytometry were used to detect apoptotic morphology and early apoptotic rate. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of miR-21-5p in AECⅡ, and Western Blot was used to detect the protein expressions of Bcl-2, Bax, and caspase-3 at the highest transfection efficiency at different time points (6, 12, 24, 48 hours).Results ① AECⅡ identification: fluorescence microscopy showed the presence of characteristic structure of AECⅡ, i.e. microvilli and osmiophilic lamellar bodies.② Apoptotic morphology: transmission electron microscopy showed cytoplasmic retraction, chromatin condensation, margination, lack of cell surface microvilli, and emptying of osmiophilic lamellar bodies in AECⅡ.③ The expression of miR-21-5p in AECⅡ: the highest transfection efficiency was found at 48 hours. The expression of miR-21-5p in miR-21-5p overexpression group was significantly higher than that of the normal control group, H2O2 challenge group and miR-21-5p negative transfection group (A value: 1.54±0.02 vs. 1.02±0.02, 0.56±0.03, 0.58±0.02, allP< 0.05).④ The rate of early apoptosis: compared with normal control group, the early apoptotic rates in H2O2 challenge group, miR-21-5p negative transfection group and miR-21-5p overexpression group were gradually elevated with the prolongation of injury time. The early apoptotic rate in miR-21-5p overexpression group was significantly lower than that of the H2O2 challenge group and miR-21-5p negative transfection group at all time points except 6 hours [12 hours: (10.73±2.80)% vs. (16.26±0.59)%, (16.04±0.70)%; 24 hours:(16.00±3.44)% vs. (23.29±2.78)%, (23.58±2.31)%; 48 hours: (31.30±3.55)% vs. (50.53±2.17)%, (49.41±1.97)%, allP< 0.05]. There was no significant difference in early apoptotic rate between miR-21-5p negative transfection group and H2O2 challenge group at each time point.⑤ Protein expression: the expressions of Bax and caspase-3 in miR-21-5p overexpression group were significantly lower than those of the H2O2 challenge group and miR-21-5p negative transfection group [Bax (A value): 0.07±0.01 vs. 0.18±0.01, 0.13±0.01; caspase-3 (A value): 0.07±0.01 vs. 0.23±0.01, 0.12±0.01, allP< 0.05], and Bcl-2 protein expression was significantly higher than that of the H2O2 challenge group and miR-21-5p negative transfection group (A value: 0.26±0.01 vs. 0.06±0.01, 0.10±0.01, both P< 0.05).Conclusions① miR-21-5p has the function of anti-apoptosis of AECⅡ.② Adenoviral vector is a successful gene transfer vector when transfected with AECⅡ.③ The anti-apoptosis of AECⅡ by miR-21-5p may be associated with reduced Bax and caspase-3 protein levels and raised expression levels of Bcl-2 protein.
6.Experimental study on induction of atherosclerotic plaque instability in rabbits after transfer of wild-type p53 gene
Wenqiang CHEN ; Yun ZHANG ; Mei ZHANG ; Xiaoping JI ; Chen LIN ; Yongfeng ZHU ; Xuedong SANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM: To study the apoptotic role of wild-type p53 in induction of plaque instability in atherosclerotic rabbits. METHODS: Fifty-four New Zealand White rabbits underwent balloon-induced abdominal aortic wall injury and then were fed on a diet of 1% cholesterol. At the end of the eighth week, the rabbits were randomly divided into two groups: group A and group B. Recombinant adenovirus carrying p53 and ?-galactosidase (LacZ) genes were injected in group A and B, respectively. Two weeks later, 10 rabbits each in group A and B was killed and the remaining rabbits all underwent pharmacological triggering with injection of Chinese Russell's viper venom and histamine. RESULTS: Compared with group B, p53 gene over-expression in group A resulted in a marked increase in number of positive apoptotic cells (2.5%?0.8% vs 1.0%?0.3%, P
7.Flavone C-glycosides from seeds of Ziziphus jujuba var. spinosa.
Ke-xian CHEN ; Li-mei ZHAO ; Chang-jiu JI ; Ning-hua TAN
China Journal of Chinese Materia Medica 2015;40(8):1503-1507
Five flavone C-glycosides were isolated from the methanol extract of the degrease seeds of Ziziphus jujuba var. spinosa though various column chromatography methods including silica gel, MPLC, and HPLC. The structures were elucidated as 6"-feruloyl- 6'''-vanillylspinosin(1), 6",6'"-diferuloylspinosin(2), spinosin(3), swertisin(4) and isoswertisin(5) based on the NMR and MS spectral data. 1 is a new compound.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Flavones
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chemistry
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isolation & purification
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Glycosides
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chemistry
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isolation & purification
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Magnetic Resonance Spectroscopy
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Mass Spectrometry
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Molecular Structure
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Seeds
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chemistry
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Ziziphus
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chemistry
8.Chemical constituents of Swertia patens.
Kang HE ; Tuan-wu CAO ; Hong-ling WANG ; Chang-an GENG ; Xue-mei ZHANG ; Ji-jun CHEN
China Journal of Chinese Materia Medica 2015;40(20):4012-4017
Chemical constituents of Swertia patens. The whole plant of air-dried Swertia patens was extracted with 90% EtOH. The water extract was suspended in H₂O and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isola- ted and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, ¹H-NMR, ¹³C- NMR). Eighteen compounds were isolated and elucidated as 3, 4-dihydro-1H,6H,8H-naptho [1,2-c:4,5-c', d'dipyrano-1, 8-dione (1), angelone (2), gentiogenal (3), erythricin (4), erythrocentaurin (5), gentianine (6), swertiakoside B (7), swertiamarin (8), 2'-O-actylswertiamarin (9), amarogentin (10), 1, 3, 5-trihydroxyxanthone (11), 1, 3-dihydroxy-5-methoxyxanthone (12), 1-hydroxy- 2, 3, 5-trimethoxyxanthone (13), gentiocrucine (14), 3-hydroxyphenylketone (15), n-hexacosyl ester 4-hydroxy-trans-cinnamate (16), n-hexacosyl ester 4-hydroxy-cis-cinnamate (17), and cholest-4-en-3-one (18). Compounds 1-7, 9-18 were obtained from S. patens for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
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Swertia
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chemistry
9.A comparative study of brain iron deposition in schizophrenia with and without tardive dyskinesia
Fangbin CHEN ; Mei JIN ; Leping XU ; Feifei ZHOU ; Li WANG ; Juying JI
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(10):916-918
ObjectiveTo explore the relationship between brain iron deposition and pathogenesis of tardive dyskinesia (TD) in schizophrenia.MethodsThe corrected phase (CP) of basal ganglia was measured in schizophrenia with TD( n=18) and without TD( n =18 ) using susceptibility weighted imaging MRI.Abnormal Involuntary Movement Scale (AIMS) was applied for clinical assessment of TD.ResultsAfter adjusting for age,sexual,and antipsychotic dosage,the mean CP of substantia nigra (SN) and caudate nucleus (CN) were significantly lower in schizophrenia patients with TD ( ( - 0.194 ± 0.040 ) rad,( - 0.089 ± 0.023 ) rad) than those without TD ( ( - 0.163 ± 0.033 ) rad,( - 0.076 ± 0.013 ) rad ; P =0.022,0.023 ).Lower mean CP in CN correlated with higher severity score of AIMS in TD patients ( r =- 0.468,P =0.034).Logistic regression analysis showed that the lower CP vaule in SN (β=-72.12,P=0.029) and CN(β=- 156.43,P=0.037),aging (β=0.379,P=0.042)were associated with the onset of TD.ConclusionThe results imply that the excess iron accumulation in basal ganglia may be associated with pathogenesis of TD in schizophrenia.
10.Activation of JAK-STAT1 signal transduction pathway in lesional skin and monocytes from patients with systemic lupus erythematosus
Jie LI ; Shuang ZHAO ; Mei YI ; Xing HU ; Ji LI ; Hongfu XIE ; Wu ZHU ; Mingliang CHEN
Journal of Central South University(Medical Sciences) 2011;36(2):109-115
Objective To study the activation of Janus protein tyrosine kinase (JAK)/signal transducer and activator of transcription 1 (STAT1) signaling pathway and its inhibitor-signal transducer and activator of transcription-1(SOCS-1) in patients with systemic lupus erythematosus. Methods A total of 45 patients with active systemic lupus erythematosus (SLE) and 30 healthy controls were randomly selected. Western blot was performed to measure the expression of Stat1 protein and phospho-Stat1 protein (an activated form of Stat1 protein) in the monocytes after stimulation with recombinant high mobility group box1 (rHMGB1) at various time points. Expression of Stat1 protein in the skin or lesional skin was also detected. Phasic expressions of SOCS-1 mRNA in the monocytes after rHMGB1 stimulation were detected by real-time reverse transcription-polymerase chain reaction. SOCS-1 gene expression in the skin or lesional skin was also detected. Results The expression level of Stat1 proteins in the monocytes from patients with SLE was higher than that from healthy controls (t=9.16,P<0.01) and positively correlated with SLE disease activity index (SLEDAI) (r=0.59,P<0.01). Expression of phospho-Stat1 in the monocytes from SLE patients was time-dependently upregulated after stimulation with rHMGB1 at various time points, while expression of SOCS-1 mRNA remained unchanged(all P>0.05). Expressions of phospho-Stat1 protein and SOCS-1 mRNA in the monocytes from healthy controls were increased transiently after stimulation with rHMGB1(all P<0.05). Both expressions of phospho-Stat1 protein and SOCS-1 gene in the lesional skin from patients with SLE were upregulated compared with those in normal skin from healthy controls (all P<0.01). Conclusion There are hyperactivation of JAK-STAT1 signaling pathway and negative feedback down-regulation of SOCS-1 in patients with systemic lupus erythematosus. HMGB-1 may be partly involved in the pathogenesis of SLE by the abnormal mediating function of JAK-STAT1 signal transduction pathway.