1.The role of phosphorylated protein kinase R-like ER kinase in brain tissue of hypoxic-ischemic neonatal rats and the following effect on neuronic apoptosis
Hui GU ; Lian JI ; Tianchu HUANG ; Yan MEI ; Zhengwei YUAN
Chinese Pediatric Emergency Medicine 2015;22(5):316-319
Objective To investigate the effect and mechanism of phosphorylated protein kinase R-like ER kinase(p-PERK) and C/EBP homologous protein(CHOP) after hypoxic-ischemic brain damage ( HIBD) . Methods Neonatal 7-day-old Sprague Dawley rats were divided into sham-operation control group and HIBD group( n=30 per group) . Each group was divided into 0 h,6 h and 24 h subgroup after operation ( n=10 per group) . The ratio of apoptosis of brain cell was measured by flow cytometer and the expression of p-PERK and CHOP were detected by Western blot. Results (1)Apoptosis cell appeared at 6 h in HIBD group,the ratio of cell apoptosis was(2. 17 ± 0. 19)%. The apoptosis cell obvious increased at 24 h,the ratio of cell apoptosis was(13. 42 ± 0. 83)%. There was a significant increase in the ratio of apoptosis after HIBD 6 h and 24 h, as compared with sham-operation control group [ ( 0. 57 ± 0. 06 )%( P <0. 01 ) ] . ( 2 ) The expression of both p-PERK and CHOP was very low in sham-operation control group. In the HIBD group,the expression of both p-PERK and CHOP began to increase at 6 h and increased furthermore at HIBD 24 h. The differences in the expression levels of p-PERK and CHOP in HIBD group among different time points were significant( P<0. 01 ) . ( 3 ) The expression of p-PERK positively correlated with the expression of CHOP (r=0. 997,P< 0. 05). Conclusion With the emerging of apoptosis after HIBD,the expression of both p-PERK and CHOP increases. The imbalance in the expression of PERK induces the apoptosis of brain cells in the HIBD of neonatal rats by regulation of CHOP expression.
2.Effect of lipoxin A4 on the Na+-K+-ATPase in alveolar type Ⅱ epithelial cells of rats treated with endotoxin
Fang CHEN ; Ru LI ; Linyan LI ; Hongxia MEI ; Qianqian HUANG ; Qingquan LIAN ; Shengwei JIN
Chinese Journal of Emergency Medicine 2010;19(12):1269-1274
Objective To study the protective role of pre-resolving mediator lipoxin A4(LXA4) in the NA+ -K+-ATPase in alveolar type Ⅱ (AT Ⅱ ) epithelial cells of rats exposed to lipopolysaccharide (LPS). Method The AT Ⅱ cells were isolated and purified, and divided randomly into control group (PBS), vehiculum (alcohol 0.7 μL/mL) group, LPS (1 μg/mL) group, LXA4(1/10 mol/mL) group and LPS (1 μg/mL LPS) + LXA4(1/10 mol/mL) group. After exposure to LPS and/or LXA4 for4 hours, NA+-K+ -ATPase and β1-subunits mRNA in AT Ⅱ epithelial cells were detected by using RT-PCR, and ATP, ADP, AMP, total adenine nucleotides (TAN) and energy charge (EC) were measured by using high performance liquid chromatography (HPLC), and then the activities of Na+-K+-ATPase were calculated accordingly. Results The NA+-K+-ATPase α-subunit and β-subunit mRNA were significantly decreased in LPS group ( P < 0.05 vs. control group). However, the expressions of NA+ -K+-ATPase mRNA were significantly enhanced by application of LXA4 to AT Ⅱ epithelial cells exposed to LPS (P <0.05 vs. LPS group). The activities of NA+ -K+ -ATPase were enhanced in LPS group (P <0.05 vs. control group). Compared with control group and LPS group, the activities of NA+-K+-ATpase in LPS + LXA4 group were significantly increased (P <0.01 vs. control group; P <0.05 vs. LPS group). The EC of AT Ⅱ epithelial cells were higher in LPS group ( P < 0.01 vs. control group). There were no significant differences in EC between control group and LPS + LXA4group(P >0.05). Conclusions The pro-resolving mediator LXA4 can enhance the expressions of NA + -K + -ATPase α-subunit and β-subunit mRNA, and the activities of NA + -K + -ATPase in AT Ⅱ epithelial cells or rats exposed to LPS, and ca also balance the metabolism of AT Ⅱ epithelial cells. These findings suggest that LXA4 plays an important role in lung edema clearance in lung injury induced by endotoxin, and the role is likely associated with the enhancement of the expressions of Na+ -K+ -AT-Pase α-subunit and β-subunit, and the activities of Na+ -K* -ATPase, maintaining the balance of metabolism of AT Ⅱ epithelial cells.
3.Progress in research on oocytes parthenogenetic activation.
Mei-lian PENG ; He-feng HUANG ; Fan JIN
Journal of Zhejiang University. Medical sciences 2007;36(3):307-312
Parthenogenetic activation is a procedure that an oocyte at meiosis II stage is activated into mitosis by some chemical or physical stimulation other than a sperm and the embryo is formed in the absence of any contribution from a male gamete. The activation of oocyte is the result of calcium ion oscillations and deactivation of some cytokines such as maturation promoting factor, mitogen-activated protein kinase and cytostatic factor. Parthenogenetic activation is artificially induced by various kinds of physical and/or chemical methods. The main activation method of human oocyte is chemical methods. The rates of activation and cleavage depend on the age, origin,and culture conditions of the oocyte.
Adenine
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analogs & derivatives
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pharmacology
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metabolism
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Cycloheximide
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pharmacology
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metabolism
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Humans
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drug effects
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growth & development
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drug effects
4.Effect of recombinant human growth hormone on final adult height in children with growth hormone deficiency
Si-Nian PAN ; Min-Lian DU ; Hong-Shan CHEN ; Yan-Hong LI ; Hua-Mei MA ; Ting-Ting HUANG ;
Chinese Journal of Endocrinology and Metabolism 2001;0(05):-
0.05).Conclusion GH improves FAH of children with GHD.Height at the initiation of puberty is the most significant determining factor for the long-term efficacy.Hence,it is important that the diagnosis should be made and treatment be initiated as early as possible to afford children with GHD the opportunity to make up much of their height deficit before puberty.Adequate dosage of GH should be used for the children taking initial treatment at puberty to attain satisfactory FAH.
6.Effect of endogenous TGF-beta1 and TNF-alpha on the As(2)O(3) inducing apoptosis of HL-60 cells.
Yuan-zhong CHEN ; Yong WU ; Mei-juan HUANG ; Lian-huang LU
Chinese Journal of Hematology 2003;24(5):231-234
OBJECTIVETo study the effect of endogenous TGF-beta(1) and TNF-alpha on As(2)O(3) inducing apoptosis of HL-60 cells.
METHODSThe expressions of endogenous TGF-beta(1) and TNF-alpha in apoptotic HL-60 cells induced by As(2)O(3) were assayed by RT-PCR, quantitative RT-PCR, ELISA, DNA fragmentation and TUNEL. The effect of TGF-beta(1) and TNF-alpha antisense phosphorothioate oligodeoxynucleotides (PSODNs) on As(2)O(3) inducing apoptotic HL-60 cells was further studied.
RESULTS(1) Expressions of endogenous TGF-beta(1) and TNF-alpha were significantly up-regulated in As(2)O(3) inducing apoptotic HL-60 cells (from 13,546 +/- 124 and 497,216 +/- 187 before treatment to 23,273 +/- 229 and 674,217 +/- 189 after treatment, respectively), accompanied with down-regulated bcl-2 mRNA expression (from 10,424 +/- 274 before treatment to 3,361 +/- 89 after treatment). (2) TGF-beta(1) and TNF-alpha antisense PSODNs could rescue As(2)O(3) induced apoptosis of HL-60 cells, with a restoration of bcl-2 gene expression.
CONCLUSIONSEndogenous TGF-beta(1) and TNF-alpha played an important role in As(2)O(3) inducing HL-60 cells apoptosis through down-regulation of bcl-2 expression.
Antineoplastic Agents ; pharmacology ; Apoptosis ; physiology ; Arsenicals ; pharmacology ; Down-Regulation ; HL-60 Cells ; Humans ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Receptors, Tumor Necrosis Factor ; metabolism ; Transforming Growth Factor beta1 ; biosynthesis ; physiology ; Tumor Necrosis Factor-alpha ; physiology
7.Small interfering RNA targeting C-erbB-2 gene increases the radiosensitivity of lung adenocarcinoma cells in vitro.
Hua-Shan SHI ; De-Lian REN ; Xin-Mei CAO ; Li HUANG ; Yao JIANG
Journal of Southern Medical University 2008;28(11):1977-1980
OBJECTIVETo investigate the effect of small interfering RNA (siRNA) targeting C-erbb-2 oncogene on the radiosensitivity of C-erbb-2-overexpressing lung adenocarcinoma cell line.
METHODSFour pairs of siRNA targeting the coding sequence of C-erbb-2 mRNA were synthesized and their interference effects were evaluated using quantitative real-time fluorescence PCR. The siRNA with the best interference effects was transfected into Calu-3 cells, which were then exposed to 2 or 5 Gy irradiation, with the cells with transfection or irradiation alone as the control groups. The cell apoptosis after the treatment was detected using annexin V-FITC Kit.
RESULTSThe apoptosis rate of the Calu cells was 7.767-/+0.551 in the blank group, 14.400-/+1.114 in the interference group, 11.867-/+0.737 in 2 Gy irradition group, 23.000-/+1.664 in 2 Gy irradiation + interference group, 16.100-/+0.624 in 5 Gy irradiation group, and 27.900-/+1.709 in 5 Gy irradiation+interference group.
CONCLUSIONThe siRNA targeting C-erbb-2 gene can enhance the radiosensitivity of Calu-3 cells to gamma-ray and increase their apoptosis rate following gamma-ray exposure in vitro.
Adenocarcinoma ; genetics ; pathology ; Apoptosis ; drug effects ; radiation effects ; Cell Line, Tumor ; Genes, erbB-2 ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Radiation Dosage ; Radiation Tolerance ; genetics
8.Detection of diarrheagenic escherichia coil harboring genomic O island 28 isolated from children diarrhea in Taiyuan.
Lian-qing LI ; Yong-feng HUANG ; Jian-rong RONG ; Su-mei WU ; Xiao-yu LIU ; Qing-yi ZHU ; Jian-guo XU
Chinese Journal of Epidemiology 2007;28(2):160-164
OBJECTIVETo investigate the etiologic value of diarrheagenic E. coil harboring genomic O island 28(OI-28) containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635), which were related to RTX (Repeat in toxin) toxin family isolated from children with diarrheal disease in Taiyuan.
METHODSIn the study, 257 fecal samples from children with diarrheal disease collected in Shanxi Children's Hospital. Diarrheagenic E. coli and enteropathogenic bacteria were isolated and identified by conventional bacterial culture and typing specific diarrheagenic E. coli (EPEC, EIEC, ETEC and EHEC) diagnostic serum, while diarrheagenic E. coli harboring genomic 01-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by PCR and DNA southern blot hybridization.
RESULTS206 strains (80.16%) of enteropathogenic bacteria were detected from 257 children with diarrhea disease, containing 149 strains (57.98%) of diarrheagenic E. coli and 57 strains(22.18%) of other entero-pathogenic bacteria. Among 3 strains (2.01%) of EPEC, 2 strains (1.34%) of ETEC, 2 strains (1.34%) EHEC were detected by typing specific serum, while all of the 142 strains (95.30%) isolated were suspected to be diarrheagenic E. coli. 21 strains (14.09%) of diarrheagenic E. coil harboring genomic O1-28 containing five putative virulence genes (Z0608, Z0609, Z0615, Z0634 and Z0635) were detected by polymerase chain reaction and DNA southen blot hybridization, 8 strains (5.37%) of diarrheagenic E. coli containing only one genomic OI-28 virulence gene, 2 strains (1.34%) of diarrheagenic E. coli containing two genomic OI-28 virulence gene. 21 children with diarrhea diseases caused OI-28-harboring E. coli containing five important putative virulence genes were among 0 to 3 years old (80.95%). These children correlating with OI-28-harboring E. coli did not present special clinical symptoms or signs.
CONCLUSIONThe diarrheagenic E. coil harboring genomic OI-28 was one of the important etiology for children with diarrheal disease in summer season.
Child ; China ; Diarrhea ; microbiology ; Escherichia coli ; genetics ; pathogenicity ; Escherichia coli Infections ; complications ; Genes, Bacterial ; Humans ; Virulence
9.Metabonomics Study on Urine 1H-NMR in Chronic Superficial Gastritis Patients with Pi-qi Deficiency Syndrome/Pi-Wei Dampness-heat Syndrome.
Xu-guang SHI ; Zhong-jie ZOU ; Mei-yin WU ; Yuan-gui ZENG ; Zhi-cheng LIAN ; Man-ting HUANG ; Meng-juan GONG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1427-1432
OBJECTIVETo observe metabolomic changes in urine of chronic superficial gastritis (CSG) patients with Pi-qi deficiency syndrome (PQDS) or Pi-Wei dampness-heat syndrome (PWDHS), thereby providing scientific evidence for syndrome typing of them.
METHODSUrine samples were collected from CSG patients with PQDS/PWDHS and healthy volunteers, 10 in each group. Proton nuclear magnetic resonance spectroscopy (1H-NMR) based metabonomic analysis was performed on urine samples. Contents of related biomarkers were analyzed by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), and urivariate statistical analysis.
RESULTSPLS-DA analysis showed that metabolites among CSG patients with PQDS/PWDHS and healthy volunteers could be mutually distinguished. Seven differentially identified metabolites were screened from urines of CSG patients with PQDS and healthy volunteers included glutamate, methionine, α-oxoglutarate, dimethylglycine, creatinine, taurine, and glucose. Four differentially identified metabolites were screened from urines of CSG patients with PWDHS and healthy volunteers included 2-hydroxybutyric acid, trimethylamine oxide, taurine, and hippuric acid. Eleven differentially identified metabolites were screened from urines of CSG patients with PQDS and PWDHS included fucose, β-hydroxybutyric acid, alanine, glutamate, methionine, succinic acid, citric acid, creatinine, glucose, hippuric acid, and lactic acid.
CONCLUSIONThe metabolic differences of CSG patients PQDS and PWDHS mainly manifested in glycometabolism, lipid metabolism, and amino acids catabolism, and 1H-NMR based metabonomics may be used in classified study of Chinese medical syndrome typing.
Biomarkers ; urine ; Discriminant Analysis ; Gastritis ; urine ; Hot Temperature ; Humans ; Hydroxybutyrates ; Ketoglutaric Acids ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; physiology ; Metabolomics ; Principal Component Analysis ; Proton Magnetic Resonance Spectroscopy ; Qi ; Syndrome
10.Clinical study of standardized three stages rehabilitation program in promoting comprehensive function in stroke patients
Ji-Rong ZHANG ; Shuang WU ; Yu HUANG ; Yan LONG ; Li-Li FENG ; Ting-Feng CHEN ; Mei LI ; Li MA ; Lian-Fang WANG ; Xiaomin FENG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(11):-
0.05).However,in the end of the treatment,the patients in the treatment group scored significantly better with all the scales than the control group(P