2.The Role of Endothelial Nitric Oxide Synthase in Endothelial Cells Apoptosis Induced by TNF-? and Angiotensin II
Ruiwei GUO ; Lixia YANG ; Chuanming GUO ; Feng QI ; Yankun SHI
Journal of Kunming Medical University 1990;0(02):-
Objective To explore the role of eNOS and NF-?B in the cultured endothelial cells apoptosis induced by TNF-? and Ang II.Methods The cultured endothelial cells were treated with TNF-? and Ang II in absence and presence of PDTC(an inhibitor of NF-?B);the mRNA of eNOS was measured by RT-PCR,the protein of eNOS and I?B? were assessed by Western-blot,the activity of NF-?B was evaluated by EMSA,and the apoptosis of cells was detected with Tunel.Results The mRNA and protein of eNOS were significantly decreased in the endothelial cells induced by TNF-? and AngII(P
3.MORPHOMETRIC ANALYSIS ON THE MITOCHONDRIA OF MYOCARDIUM IN EXERCISE TRAINED RATS
Qi FENG ; Shi CHENG ; Yunzong PU ; Baohui ZHANG ;
Chinese Journal of Sports Medicine 1983;0(03):-
Adult male rats were divided at random into two groups and trained on the treadmill running schedules, 60 min/day and 10min/day for group A and B separately. Four weeks later the ultrastructure of the left ventricular myocardium was examined and the specific surface of mitochondria wasmeasured. Comparing to the corresponding control group, the specific surface of mitochondria in the two groups of trained rats was decreased significantly. (5.883+0.12 um and 6.050?0.12 um to 6.564?0.17 um). This means myocardium mitochondria in trained rats were swelling. The reason might be that the myocardium injury was induced by exercise training.
4.Effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis by inhibiting AKT, ERK and upstream signal SRC activity.
Shi-mei QI ; Jun LV ; Yu MENG ; Zhi-lin QI ; Lie-feng LING
China Journal of Chinese Materia Medica 2015;40(16):3267-3272
To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic
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pharmacology
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Apoptosis
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drug effects
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Breast Neoplasms
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drug therapy
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enzymology
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genetics
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physiopathology
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Down-Regulation
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Extracellular Signal-Regulated MAP Kinases
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genetics
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metabolism
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Female
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Humans
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MCF-7 Cells
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Proto-Oncogene Proteins c-akt
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genetics
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metabolism
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Saponins
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pharmacology
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Signal Transduction
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drug effects
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Triterpenes
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pharmacology
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src-Family Kinases
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genetics
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metabolism
5.The research and strategy about the current situation about the analysis of cellular morphology in Tibet region
Quangui SHI ; Dongfang FENG ; Sijuan DING ; Hongzhang CHEN ; Baohong QI ; Huaying SHI
International Journal of Laboratory Medicine 2014;(8):1007-1008
Objective To understand the current situation and level of clinical laboratory about the analysis of cellular morphology in Tibet region .Methods Authors investigated the information about the staff of clinical laboratory testing the patient′s blood smear under microscope ,executing the rules and regulations by using standard questionnaires .Results Some of the clinical laboratory didn′t founded the rule and standardization of rechecking about abnormal blood routine (5/15 ,33 .3% ) .Some of the divi-sion leadership didn′t pay enough attention to the staff′s basic operation (2/15 ,13 .3% ) .Most of the staff didn′t being trained about cellular morphology in special purpose workshop (6/43 ,88 .8% ) .Some of the hospital didn′t carried out the chemistry stai-ning about blood cells(10/15 ,66 .7% ) .Conclusion It′s important to promote the quality and level about the analysis of cellular morphology in Tibet region .
6.Changes of HCN4, Cx43 Expression in the Sinoatrial Node of Electric Shock Death.
Xiao-feng CHEN ; Dong LIANG ; Qi HAN ; Shi-feng ZHOU ; Mao-jin ZHENG ; Chao-qun WANG
Journal of Forensic Medicine 2015;31(4):266-268
OBJECTIVE:
To investigate the expression of hyperpolarization-activated cyclic nucleotide-gated cation channel 4 (HCN4) and connexin43 (Cx43) in the sinoatrial node of electric shock death.
METHODS:
As experimental group, 34 cases of electric shock death who had definite current mark evidence were selected from pathology department of Xuzhou Medical College from 2010 to 2013. As the control group, 20 cases of fatal severe craniocerebral injury in traffic accidents were chosen. The expressions of HCN4 and Cx43 in the sinoatrial node were observed by immunohistochemical technology.
RESULTS:
HCN4 positive cells expressed in the cell membrane and cytoplasm of the sinoatrial node. Cx43 positive cells expressed in the cell membrane and cytoplasm of T cells and myocardial cells. The expression of HCN4 was significantly higher than that of the control group (P < 0.05) and the expression of Cx43 was significantly lower than that of the control group (P < 0.05).
CONCLUSION
The changes of HCN4 and Cx43 expressions in the sinoatrial node illustrate electric shock death might be related to the abnormalities of cardiac electrophysiology and conduction.
Connexin 43/metabolism*
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Cyclic Nucleotide-Gated Cation Channels
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Heart Rate
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Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/metabolism*
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Immunohistochemistry/methods*
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Myocardium/metabolism*
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Myocytes, Cardiac
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Sinoatrial Node/physiopathology*
7.Quality standard study on Mori Cortex liquid extract.
Mao-feng LIU ; Mei-ping XIE ; Lan LI ; An-qi LU ; Jian-gong SHI ; Su-juan WANG
China Journal of Chinese Materia Medica 2015;40(10):1850-1854
A reasonable and practicable quality standard was developed for mori liquid extract from different sources by TLC, HPLC and fingerprint technology. In TLC method, the compounds were separated on polyamide film using glacial acetic acid-water (1: 3) as mobile phase at a UV wavelength of 365 nm. All qualified samples had the spots of the same color as the control herb and substance. The RP-HPLC method was used to determine the content of mulberroside A with mobile phase of methanol-water (25: 75) at a wave-length of 326 nm. The mulberroside A was in good linear with a regression equation of Y = 46.965X (r = 0.999 6) in the range of 4.6 - 228 mg x L(-1). In 14 batches of samples, the mulberroside A in 4 batches of them was less than 0.5 g x L(-1), and was more than 2.0 g x L(-1) in the other batches. It was suggested that the content limit of mulberroside A should be no less than 1.5 g x L(-1). The HPLC fingerprints were evaluated by the similarities. It has found that the similarities of different mori liquid extracts were very low and the chemical diversity of mori cortex was the major factor of similarity. Moreover, the process impact was minimal. Thus the fingerprint was not included in this quality standard.
China
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Chromatography, High Pressure Liquid
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Disaccharides
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chemistry
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isolation & purification
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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standards
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Morus
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chemistry
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Quality Control
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Stilbenes
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chemistry
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isolation & purification
8.Expression of BSAP/CD30 in classic Hodgkin lymphoma using double-staining technique.
Yan-Feng XI ; Wen-Qi BAI ; Jin-Fen WANG ; Quan-Hong WANG ; Shi-Lan JIAO
Chinese Journal of Pathology 2007;36(2):136-137
Adolescent
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Adult
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Aged
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Biomarkers, Tumor
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metabolism
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Child
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Female
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Gene Expression Regulation, Neoplastic
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Hodgkin Disease
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genetics
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metabolism
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Humans
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Ki-1 Antigen
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metabolism
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Male
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Middle Aged
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PAX5 Transcription Factor
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metabolism
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Staining and Labeling
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methods
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Young Adult
9.Activated peroxisome proliferator-activated receptor αinhibiting the lipopolysaccharide-induced macrophage-mediated inflammatory responses by promoting the autophagy
Rongrong YANG ; Li ZHANG ; Xiangying ZHANG ; Hongbo SHI ; Dexi CHEN ; Zhongping DUAN ; Feng REN ; Qi WANG
Chinese Journal of Microbiology and Immunology 2015;(6):431-435
Objective To investigate the effects of peroxisome proliferator-activated receptor α( PPARα) on macrophage-mediated inflammatory responses with the interference of lipopolysaccharide and the possible mechanism.Methods The bone marrow stem cells were isolated from the femora of mice.The granulocyte-macrophage colony stimulating factor ( GM-CSF) was used to stimulate the in vitro differentiation from bone marrow stem cells into primary macrophages.An in vitro model with cultured cells expressing in-flammatory cytokines was established by treating the primary macrophages with lipopolysaccharide ( LPS) .A specific chemical agonist, Wy-14643, was used to activate PPARα. Autophagy inhibitors including 3-methyladenine (3-MA) and small interfering RNA against Atg7 ( Atg7 siRNA) were used to inhibit the autophagy.Western blot assay was performed to detect the expression of autophagy-related proteins ( Atg5, Atg7, Beclin-1 and LC3).The transcriptional levels of TNF-α, IL-1β, IL-6, Atg5, Atg7 and Beclin-1 were analyzed by qRT-PCR.Results Compared with the macrophages treated with LPS alone, those pretreated with various concentrations of Wy-14643 (10 μmol/L, 25 μmol/L and 50 μmol/L) showed inhibited ex-pression of proinflammatory cytokines ( TNF-α,IL-1βand IL-6) and enhanced expression of autophagy-relat-ed proteins (Atg5, Atg7 and Beclin-1) at mRNA level in a dose-dependent manner.The expression of auto-phagy-related proteins (Atg5, Atg7, Beclin-1 and LC3) by macrophages was promoted with the pretreatment of Wy-14643 as indicated by Western blot assay.The transcriptional levels of TNF-α, IL-1βand IL-6 were increased in Wy-14643 pretreated-macrophages after stimulation with 3-MA or Atg7 siRNA .Conclusion PPARαsuppressed the macrophage-mediated inflammatory responses by promoting autophagy, suggesting that the PPARα-autophagy pathway might be one of the signaling pathways regulating LPS induced-inflamma-tory responses.
10.Caspase-1 aggravates the D-galactosamine/lipopolysaccharide induced acute liver failure in mice through activating glycogen synthase kinase-3β
Rongrong YANG ; Feng REN ; Li ZHANG ; Xiangying ZHANG ; Hongbo SHI ; Dexi CHEN ; Zhongping DUAN ; Qi WANG
Chinese Journal of Microbiology and Immunology 2016;36(2):132-137
Objective To analyze the role of cysteinyl aspartate specific proteinase-1 (caspase-1) in a mouse model of D-galactosamine (D-GalN) and lipopolysaccharide (LPS) induced acute liver failure (ALF) and to study the possible mechanism. Methods C57BL/ 6 mice were randomly divided into four groups including control group, Z-WEHD-FMK (caspase-1 inhibitor) treatment group, ALF model group and Z-WEHD-FMK-treated ALF group. The mouse model of ALF was established by intraperitoneally injec-ting the mice with D-GalN (450 mg/ kg) and LPS (10 μg/ kg). The damages in liver tissues were evaluated based on the histopathological examination and the levels of alanine transaminase (ALT) and aspartate trans-aminase (AST) in serum samples. Western blot assay was performed to analyze the expression of caspase-1 and the phosphorylation of glycogen synthase kinase 3β (GSK-3β). The qRT-PCR was used to measure the expression of inflammatory cytokines at transcriptional level. Results The expression of caspase-1 at both mRNA and protein levels were gradually increased during the development of ALF. Compared with the mice with ALF, those in the Z-WEHD-FMK-treated ALF group showed less severe liver damages on histopatholog-ical examination and decreased levels of ALT and AST in serum samples [ALT: (479. 2±39. 5) U/ L vs (998. 5±60. 4 ) U/ L, P<0. 05; AST: ( 478. 5±28. 6) U/ L vs ( 1 180. 7±91. 4) U/ L, P<0. 05]. The expression of TNF-α, IL-1β, IL-18 and IL-33 at transcriptional level were significantly suppressed in mice with ALF upon the Z-WEHD-FMK intervention. Results of the Western blot assay indicated that Z-WEHD-FMK suppressed the activities of GSK-3β by enhancing its phosphorylation. Conclusion This study demon-strated that caspase-1 could promote the activation of GSK-3β resulting in the development of inflammation responses and liver damages during the development of ALF in mice.