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1.
Journal of Medical Postgraduates ; (12): 466-470, 2020.
Article in Chinese | WPRIM | ID: wpr-821875

ABSTRACT

ObjectiveThe mechanism of histone phosphorylation modification in oocyte meiosis is less studied. This study is designed to investigate the pattern of histone H3 phophorylation and regulation of maturation process in the porcine oocytes.MethodsThe histone H3Ser10 (H3S10) phosphorylation expression was examined on the porcine oocyte meiotic process. The porcine cumulus oocyte complexes (COCs) were divided into four groups, one group was cultured as control group, and the other 3 groups were supplemented with 5, 10, and 30 μmol/L ZM447439, and cultured in vitro for 27 h, respectively, 5, 10, and 30 μmol/L ZM4474349 treatment group. The proportion of each meiotic stage was counted. The phosphorylation pattern of histone H3S10 and the expression level of protein kinase Aurora B were detected at the porcine oocytes.ResultsCompared with histone H3S10 phosphorylation level of oocyte GVBD phase, the MI and AI phases were significantly increased (P<0.05), and H3S10 phosphorylation level of AI phase was remarkedly higher than that of MII phase (P<0.05). Compared with the control group, the proportion of oocytes at the GVBD phase in the 10 and 30 μmol/L ZM4447439 treatment group [(32.14±0.51)%, (95.34±0.59)%]was higher than that of the control group [(2.56±0.03)%, P<0.05], the proportion of oocytes at the MI phase [(66.88±0.13)%, (4.66±0.04)%] significantly decreased than that of the control group [(87.42±0.14)%, P<0.05], and the proportion of oocytes at the AI stage [(1.01±0.03)%, (0.000±0.00)%] significantly decreased compared with the control group[(10.02 ± 0.21)%, P<0.05]. Compared with the control group (0), oocytes H3S10 dephosphorylation modification ratio in the 10 μmol/L and 30 μmol/L ZM4474349 treatment group [(35.2±0.39)%, (95.4±0.65)%]significantly increased (P<0.05). Compared with the control group, the relative expression level of Aurora B in the 10 and 30 μmol/L ZM4447439 treatment group was significantly reduced (P<0.05).ConclusionHstone H3S10 phosphorylation plays arolein the maturation of mammalian oocytes. AuroraB kinase inhibitors (ZM447439) treatment can reduce H3S10 phosphorylation and Aurora B expression level and lead to oocytesmaturation disorder.

2.
Article in Chinese | WPRIM | ID: wpr-755572

ABSTRACT

Objective To evaluate the relationship between mechanical ventilation-induced apoptosis in hippocampal neurons and mammalian taget of rapamycin ( mTOR) signaling pathway in mice. Methods Fifty healthy male C57BL∕6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 2 groups ( n=25 each) using a random number table method: control group ( group C ) and mechanical ventilation group ( group V) . The mice breathed spontaneously for 6 h in group C, and the mice were mechanically ventilated for 6 h in group V. Open field test and contextual fear conditioning test were conducted at 1 and 3 days after the end of ventilation. Hippocampal tissues were obtained at 1 day after the end of ventilation for determina-tion of the expression of mTOR, phosphorylated mTOR (p-mTOR), microtubule-associated protein 1 light chain 3Ⅱ( by Western blot) and apoptosis in hippocampal neurons ( by TUNEL) . The p-mTOR∕mTOR ratio and apoptosis index were calculated. Results Compared with group C, the time animals spent in the central square was significantly prolonged, the number of crossing the grid was reduced, the percentage of freezing time was decreased, the expression of microtubule-associated protein 1 light chain 3Ⅱwas up-regulated, and the p-mTOR∕mTOR ratio and apoptosis index were increased in group V ( P<0. 05) . Conclusion The mech-anism by which mechanical ventilation induces apoptosis in hippocampal neurons may be related to activation of mTOR signaling pathway in mice.

3.
Acta Pharmaceutica Sinica ; (12): 454-462, 2019.
Article in Chinese | WPRIM | ID: wpr-780137

ABSTRACT

The purpose of this study is to further explore the effects of SI-4650, a newly discovered small molecule inhibitor of spermine oxidase (SMO) in our laboratory, on proliferation and migration of human osteosarcoma 143B cells and its underlying molecular mechanism. Chemiluminescence and high performance liquid chromatograph were used to analyze the effect of SI-4650 on SMO activity in 143B cells. DCFH-DA-staining/FCM was used to analyze the accumulation of cellular reactive oxygen species (ROS), whereas MTT and FCM were used to detect proliferation and cell cycle. Transwell culture and Western blot were used to analyze the expression levels of migration-related proteins. PI/FITC-Annexin V/FCM, fluorescence microscopy and Western blot were used to analyze apoptosis and autophagy. Our results showed that SI-4650 could significantly decrease SMO activity, inhibit cell proliferation or migration, and induce a S-phase cell cycle arrest in 143B human osteosarcoma cells. The mechanism may be related to interfering with polyamine metabolism, activating mitochondrial-mediated apoptosis and causing autophagic death. These results suggest that SI-4650 has the potential for clinical use in treatment of osteosarcoma.

4.
Article in Chinese | WPRIM | ID: wpr-695704

ABSTRACT

Objective· To investigate the correlation between increased nuchal translucency (NT) thickness and fetal chromosomal abnormalities,severe structure anomalies and other abnormalities,and its value in assessment of fetal prognosis.Methods · Five hundred and eighty-three singleton fetuses with NT ≥ 2.5 mm in the first trimester (11-13+6 week) were retrospectively analyzed,of which 252 had invasive prenatal test for fetal chromosome and genetic tests.They were divided into 5 groups according to the NT thickness,2.5 ~ 2.9 mm,3.0 ~ 3.4 mm,3.5 ~ 4.4 mm,and ≥ 4.5 mm as 1st to 4th group,and cystic hygroma as 5th group.The incidences among groups were analyzed by Cochran-Armitage Trend test.Fisher's exact test was used to compare diversities of NT thickness among the major chromosomal abnormalities.Results · Among the 583 singleton fetuses,59 were diagnosed as chromosomal abnormalities (23.4%,59/252),38 with structure anomalies (6.5%),of which 13 cases with severe cardiac anomalies (2.2%).There were 6 fetal demise,3 inevitable abortion,2 stillborn,94 terminations of pregnancy (8 for personal factors) and 478 live birth,without spontaneous abortions and congenital infections after invasive prenatal test.The differences among the incidences of chromosomal abnormalities,structure anomalies and cardiac anomalies in five groups were statistically significant (P=0.000) and the incidences all increased with fetal NT thickness.The healthy living rates of fetus were 96.5%,81.9%,74.0%,35.6%,and 6.7% among groups,respectively,and the incidences all decreased with fetal NT thickness (P=0.000).Conclusion· Increased NT thickness is related to fetal chromosomal abnormalities,severe cardiac anomalies and poor pregnant outcome.The incidences of chromosomal abnormalities,structure anomalies,cardiac anomalies and pregnant outcome all increase with fetal NT thickness.In clinical practice,individualized guidance should be conducted according to different thickness of NT.

5.
Basic & Clinical Medicine ; (12): 475-479, 2018.
Article in Chinese | WPRIM | ID: wpr-693925

ABSTRACT

Objective To evaluate whether down-regulating antizyme inhibitor(AZIN) can regulate the expression of ornithine decarboxylase(ODC) and the proliferation of prostate cancer cell PC3 or not.Methods siRNA-AZIN transfected prostate cancer cell PC3,the level of antizyme(AZ),AZIN and ODC were measured by RT-PCR and westernblot. MTT was used to measure the proliferation of cells. Results The mRNA level of AZIN declined(P<0.01);the protein level of AZIN and ODC declined(P<0.05). Knockdown of AZIN significantly inhibited the proliferation of PC3(P<0.05). Conclusions Transfecting siRNA-AZIN can decrease the level of AZIN, then the decline level of ODC inhibits the proliferation of PC3.

6.
Article in Chinese | WPRIM | ID: wpr-737252

ABSTRACT

The aim of the present study was to investigate the effect of lipoxin A4 (LXA4)pretreatment on cognitive function of aged rats after global cerebral ischemia reperfusion,and to explore its possible mechanism.Thirty-six aged male Sprague-Dawley rats were randomly divided into three groups (n=12 each):sham-operation group (S group),global cerebral ischemia reperfusion group (I/R group) and LXA4-pretreatment group (L group).The rat model of global cerebral ischemia reperfusion was established by occlusion of the bilateral common carotid artery with hypotension.The cognitive function of rats was determined by a step-down type passive avoidance test and Morris Water Maze test on the third day after reperfusion.Rats were sacrificed after Water Maze test and the pathological changes ofhippocampal CA1 region were observed and the related inflammatory mediators were determined.As compared with S group,the escape latency in I/R group was prolonged from the first day to the fifth day,while that in L group was prolonged from the first day to the third day.The retention time in I/R group and L group in the first quadrant was shortened.The reaction time,frequency of reaction mistake and frequency of escape mistake in I/R group increased,and the latent period shortened.The frequency of escape mistake in L group increased,and the damage in the hippocampal CA1 region of I/R group and L group was obvious.The levels of S-100β,TNF-α,IL-lβ,IL-10 and NF-κB in I/R group and L group increased.As compared with I/R group,the escape latency in L group was shortened from the first day to the fifth day,and the retention time in the first quadrant prolonged.The reaction time,frequency of reaction mistake and frequency of escape mistake in L group decreased,and the latent period prolonged.The damage in the hippocampal CA1 region of L group was alleviated as well.The levels of S-100β,TNF-α,IL-1β and NF-κB in L group decreased,and those of IL-10 increased.It can be concluded that LXA4 pretreatment can improve the cognitive function in aged rats after global cerebral ischemia reperfusion probably by inhibiting the inflammatory reaction.

7.
Article in Chinese | WPRIM | ID: wpr-735784

ABSTRACT

The aim of the present study was to investigate the effect of lipoxin A4 (LXA4)pretreatment on cognitive function of aged rats after global cerebral ischemia reperfusion,and to explore its possible mechanism.Thirty-six aged male Sprague-Dawley rats were randomly divided into three groups (n=12 each):sham-operation group (S group),global cerebral ischemia reperfusion group (I/R group) and LXA4-pretreatment group (L group).The rat model of global cerebral ischemia reperfusion was established by occlusion of the bilateral common carotid artery with hypotension.The cognitive function of rats was determined by a step-down type passive avoidance test and Morris Water Maze test on the third day after reperfusion.Rats were sacrificed after Water Maze test and the pathological changes ofhippocampal CA1 region were observed and the related inflammatory mediators were determined.As compared with S group,the escape latency in I/R group was prolonged from the first day to the fifth day,while that in L group was prolonged from the first day to the third day.The retention time in I/R group and L group in the first quadrant was shortened.The reaction time,frequency of reaction mistake and frequency of escape mistake in I/R group increased,and the latent period shortened.The frequency of escape mistake in L group increased,and the damage in the hippocampal CA1 region of I/R group and L group was obvious.The levels of S-100β,TNF-α,IL-lβ,IL-10 and NF-κB in I/R group and L group increased.As compared with I/R group,the escape latency in L group was shortened from the first day to the fifth day,and the retention time in the first quadrant prolonged.The reaction time,frequency of reaction mistake and frequency of escape mistake in L group decreased,and the latent period prolonged.The damage in the hippocampal CA1 region of L group was alleviated as well.The levels of S-100β,TNF-α,IL-1β and NF-κB in L group decreased,and those of IL-10 increased.It can be concluded that LXA4 pretreatment can improve the cognitive function in aged rats after global cerebral ischemia reperfusion probably by inhibiting the inflammatory reaction.

8.
Article in Chinese | WPRIM | ID: wpr-271914

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the efficiency of direct fluorescence in situ hybridization (D-FISH) versus FISH on CD138 immunomagnetic sorting myeloma cells (MACS-FISH) to detect the cytogenetic abnormalities of multiple myeloma.</p><p><b>METHODS</b>Thirty-one patients with multiple myeloma (MM) were detected by D-FISH and MACS-FISH, using 5 probes, including 1q21, D13S319, RB1, IgH, P53. The IgH rearrangement positive patients were further examined by 3 IgH rearrangement subtype FISH probes including IgH/FGFR3, IgH/MAF and IgH/CCND1.</p><p><b>RESULTS</b>Metaphase karyotyping revealed cytogenetic abnormalities in 5 cases (16.1%), clonal aberrations were detected in 13 cases(41.9%) by D-FISH, while 25 case(80.6%) with clonal aberrations by MACS-FISH. The results between these 2 FISH methods were significantly different (P=0.042). The detection frequency of clonal aberration by each probes of D-FISH was 22.6%,25.8%,29%,38.7% and 9.7% respectively for 1q21 amplification, D13S319 deletion,RB1 deletion, IgH rearrangement and P53 deletion, compared with 48.4%,45.2%,48.4%,67.7% and 16.1% respectively by MACS-FISH. The 2 FISH methods were well consistent when the percentage of plasma cells was ≥20% in bone marrow smears. When the percentage of plasma cells was<20% in bone marrow smears, the difference between these 2 methods was very statistically significant (P=0.00).</p><p><b>CONCLUSION</b>MACS-FISH can obviously improve the detection efficiency of cytogenetic abnormalities in patients with MM. Conventional cytogenetics combined with MACS-FISH is an ideal efficient method to detect the cytogenetic abnormalities in MM patients, and should be applied widely, especially for those patients with the plasma cells <20% in bone marrow smears.</p>

9.
National Journal of Andrology ; (12): 873-877, 2017.
Article in Chinese | WPRIM | ID: wpr-812865

ABSTRACT

Objective@#To investigate the role of pH2AX in the reversibility of mouse testicular reproductive function impaired by single heat stress.@*METHODS@#Twenty-four C57 male mice were randomly divided into heat stress and control groups and immersed in water at 43℃ and 25℃, respectively, for 15 minutes. At 1, 7, and 14 days of heat exposure, all the mice were sacrificed and their testis tissues collected for determining the apoptosis of the germ cells by TUNEL and measuring the expression level of the pH2AX protein by immunohistochemistry and Western blot.@*RESULTS@#The highest percentage of apoptotic cells were found in the seminiferous tubules of the mice in the heat stress group on the 1st day of the exposure and almost no apoptosis was observed at 7 and 14 days. The pH2AX protein was expressed in the nuclei of the basement membrane of adjacent seminiferous tubules. Compared with the control group, the expression of pH2AX was significantly increased on the 1st day of exposure (0.47 ± 0.02 vs 1.61 ± 0.04, P <0.01), then decreased at 7 days (0.85 ± 0.03) in comparison with that on the 1st day (P <0.01), and again elevated at 14 days (1.72 ± 0.02) as compared with either those at 1 and 7 days (P <0.01) or that of the control (P <0.01).@*CONCLUSIONS@#Heat stress causes dynamic changes of the pH2AX expression in the testis of the mouse, which are associated with heat stress-induced proliferation and division of the testicular spermatogenic cells.


Subject(s)
Animals , Apoptosis , Blotting, Western , Heat Stress Disorders , Histones , Metabolism , Hot Temperature , Immunohistochemistry , In Situ Nick-End Labeling , Male , Mice , Mice, Inbred C57BL , Random Allocation , Seminiferous Tubules , Cell Biology , Spermatozoa , Cell Biology , Metabolism , Testis , Time Factors
10.
Article in English | WPRIM | ID: wpr-636698

ABSTRACT

The effect of the complement C1q expression on total hepatic ischemia-reperfusion (I/R) injury in rats was investigated. Sixty healthy male Sprague Dawley (SD) rats weighing 180-200 g were randomly divided into 5 groups: sham-operation group (S group, n=12); group of I/R for 1 h (I/R 1 h group, n=12); group of I/R for 3 h (I/R 3 h group, n=12); group of I/R for 6 h (I/R 6 h group, n=12); group of I/R for 24 h (I/R 24 h group, n=12). The hepatic I/R model of rats was established, and liver tissues were obtained 1 h, 3 h, 6 h and 24 h after hepatic I/R, respectively. Furthermore, the tissues were stained using hematoxylin-eosin, and the liver injuries of rats were observed using a microscope. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in liver tissue were determined. Real-time polymerase chain reaction (PCR) and Western blotting were used to detect the expression levels of C1q mRNA and protein, respectively. As compared with the S group, the histopathological changes in I/R 1 h-24 h groups were gradually aggravated with the extension of I/R time. As compared with the S group, SOD activity and MDA content in the I/R groups were reduced and increased respectively with the extension of I/R time (P<0.01). Furthermore, the C1q expression at mRNA and protein levels in the I/R groups (especially in the I/R 3 h group) was significantly higher than that in the S group (P<0.05). It is suggested that C1q expression may play a principal role in hepatic I/R injury, particularly at the early stage of perfusion.

11.
Article in English | WPRIM | ID: wpr-351063

ABSTRACT

The effect of the complement C1q expression on total hepatic ischemia-reperfusion (I/R) injury in rats was investigated. Sixty healthy male Sprague Dawley (SD) rats weighing 180-200 g were randomly divided into 5 groups: sham-operation group (S group, n=12); group of I/R for 1 h (I/R 1 h group, n=12); group of I/R for 3 h (I/R 3 h group, n=12); group of I/R for 6 h (I/R 6 h group, n=12); group of I/R for 24 h (I/R 24 h group, n=12). The hepatic I/R model of rats was established, and liver tissues were obtained 1 h, 3 h, 6 h and 24 h after hepatic I/R, respectively. Furthermore, the tissues were stained using hematoxylin-eosin, and the liver injuries of rats were observed using a microscope. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity in liver tissue were determined. Real-time polymerase chain reaction (PCR) and Western blotting were used to detect the expression levels of C1q mRNA and protein, respectively. As compared with the S group, the histopathological changes in I/R 1 h-24 h groups were gradually aggravated with the extension of I/R time. As compared with the S group, SOD activity and MDA content in the I/R groups were reduced and increased respectively with the extension of I/R time (P<0.01). Furthermore, the C1q expression at mRNA and protein levels in the I/R groups (especially in the I/R 3 h group) was significantly higher than that in the S group (P<0.05). It is suggested that C1q expression may play a principal role in hepatic I/R injury, particularly at the early stage of perfusion.


Subject(s)
Animals , Blotting, Western , Complement C1q , Genetics , Metabolism , Gene Expression , Liver , Metabolism , Male , Malondialdehyde , Metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , Reperfusion Injury , Reverse Transcriptase Polymerase Chain Reaction , Superoxide Dismutase , Metabolism , Time Factors
12.
National Journal of Andrology ; (12): 501-505, 2013.
Article in Chinese | WPRIM | ID: wpr-350872

ABSTRACT

<p><b>OBJECTIVE</b>To study the inhibitory effect of Akt inhibitor deguelin on PC-3 human prostate cancer cell lines and its possible mechanism.</p><p><b>METHODS</b>PC-3 human prostate cancer cells were cultured in deguelin at the concentrations of 10, 100, 500 and 1 000 nmol/L for 24, 48 and 72 hours, respectively. Then the inhibitory effect of deguelin on the proliferation of the PC-3 cells was determined by MTT assay and that on the cell cycle was detected by flow cytometry. The expression levels of MDM2 and GSK3beta mRNA were measured by RT-PCR and those of MDM2 and GSK3beta proteins by Western blot.</p><p><b>RESULTS</b>At 24, 48 and 72 hours, the inhibition rates of deguelin on the proliferation of the PC-3 prostate cancer cells were (91.10 +/- 3.75), (86.39 +/- 1.16) and (79.51 +/- 2.63)% at 10 nmol/L, (82.46 +/- 3.65), (76.84 +/- 0.97) and (69.69 +/- 2.30) % at 100 nmol/L, (81.46 +/- 0.41), (75.56 +/- 1.12) and (54.07 +/- 3.21)% at 500 nmol/L, and (66.77 +/- 2.82), (58.22 +/- 0.35) and (39.34 +/- 2.40)% at 1000 nmol/L, all with statistically significant differences from the control group (P < 0.01). Deguelin at 10, 100, 500 and 1 000 nmol/L increased the cell cycles blocked in the G0/G1 phase ([62.4 +/- 2.2], [63.6 +/- 1.1 ], [65.0 +/- 0.3] and [66.5 +/- 1.9]%, P < 0.01) and reduced the percentage of the S-phase cells ([14.7 +/- 2.4], [11.1 +/- 5.2], [5.8 +/- 1.1] and [7.0 +/- 0.6]%, P < 0.01). RT-PCR and Western blot showed markedly up-regulated expressions of GSK3 P3 a3beta down-regulated expressions of MDM2 mRNA and proteins in the PC-3 cells treated with deguelin.</p><p><b>CONCLUSION</b>Akt inhibitor deguelin can inhibit the proliferation of PC-3 human prostate cancer cells by affecting the down-stream signal molecules GSK3P3 and betaDM2 in the Akt pathway.</p>


Subject(s)
Cell Line, Tumor , Cell Proliferation , Glycogen Synthase Kinase 3 , Metabolism , Glycogen Synthase Kinase 3 beta , Humans , Male , Prostatic Neoplasms , Metabolism , Proto-Oncogene Proteins c-akt , Proto-Oncogene Proteins c-mdm2 , Metabolism , Rotenone , Pharmacology
13.
Article in English | WPRIM | ID: wpr-239810

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the protective effects of 8% emulsified isoflurane after myocardial ischemia-reperfusion injury and its mechanism in rabbits.</p><p><b>METHODS</b>Twenty-four male adult New Zealand white rabbits were anesthetized with intravenous injection of 30 mg/kg pentobarbital followed by 5 mg x kg(-1) x h(-1) infusion. All rabbits were subjected to 30 minutes of left anterior descending coronary artery (LAD) occlusion and 3 hours of subsequent reperfusion. Before LAD occlusion, the rabbits were randomly allocated into three groups for preconditioning treatment (eight for each group). The control group (C group) received intravenously 0.9% NaCl for 30 minutes. The emulsified isoflurane group (EI group) received 8% emulsified isoflurane intravenously till 0.64% end-tidal concentration for 30 minutes that was followed by a 15-minute washout period. The Intralipid group (IN group) received 30% Intralipid for 30 minutes. The infarcted area, plasma malondialdehyde (MDA) content, superoxide dismutase activity (SOD) and nitrite concentration after 3-hour myocardial perfusion were recorded simultaneously.</p><p><b>RESULTS</b>For the myocardial ischemia-reperfusion injury animals, the infarcted size in the EI group was significantly reduced (91.9% +/- 8%) as compared with control group (39% +/- 6%, t=5.19, P<0.01). The plasma SOD activity and nitrite concentration in EI group were significantly higher than those in control group (t=2.82, t=8.46, P<0.05), but MDA content was lower in EI group than that in control group (t=2.56, P<0.05).</p><p><b>CONCLUSIONS</b>The results indicate that emulsified isoflurane has a cardioprotection effect against ischemia-reperfusion injury. This beneficial effect of emulsified isoflurane is probably through NO release and consequently by increase in antioxidation of myocardium.</p>


Subject(s)
Animals , Emulsions , Isoflurane , Pharmacology , Lipid Peroxidation , Male , Myocardial Infarction , Drug Therapy , Pathology , Myocardial Reperfusion Injury , Nitric Oxide , Blood , Rabbits , Superoxide Dismutase , Metabolism
14.
Chinese Medical Journal ; (24): 1394-1399, 2009.
Article in English | WPRIM | ID: wpr-292703

ABSTRACT

<p><b>BACKGROUND</b>The critical roles of polyamines in cell growth and differentiation have made polyamine metabolic pathway a promising target for antitumor therapy. Recent studies have demonstrated in vitro that some antitumor polyamine analogues could be used as substrates and oxidized by purified recombinant human N(1)-acetylpolyamine oxidase (APAO, an enzyme that catabolizes natural polyamines), indicating a potential role of APAO in determining the sensitivity of cancer cells to specific antitumor analogues. This study evaluated, in vivo, the effect of APAO on cytotoxicity of antitumor polyamine analogue, N(1)-cyclopropylmethyl-N(11)-ethylnorspermine (CPENS) and its mechanism when highly expressed in human lung cancer line A549.</p><p><b>METHODS</b>A clone with high expression of APAO was obtained by transfecting A549 lung cancer cell line with pcDNA3.1/APAO plasmid and selecting with quantitative realtime PCR and APAO activity assay. Cell proliferation was determined by MTT method and apoptosis related events were evaluated by DNA fragmentation, sub-G1/flow cytometric assay, western blotting (for cytochrome C and Bax) and colorimetric assay (for casapse-3 activity).</p><p><b>RESULTS</b>A clone highly expressing APAO was obtained. High expression of APAO in A549 cells inhibited accumulation of CPENS, decreased their sensitivity to the toxicity of CPENS and prevented CPENS induced apoptosis.</p><p><b>CONCLUSION</b>These results indicate a new drug resisting, mechanism in the tumor cells. High expression of APAO can greatly decrease the sensitivity of tumor cells to the specific polyamine analogues by detoxifying those analogues and prevent analogue induced apoptosis.</p>


Subject(s)
Apoptosis , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Flow Cytometry , Humans , Oxidoreductases Acting on CH-NH Group Donors , Genetics , Metabolism , Polyamines , Metabolism , Pharmacology , Reverse Transcriptase Polymerase Chain Reaction
15.
Saudi Medical Journal. 2008; 29 (3): 368-373
in English | IMEMR | ID: emr-90139

ABSTRACT

To evaluate the protective effect of urinary trypsin inhibitor UTI on acute lung and liver injury in rat model induced by sepsis with infra-abdominal infection. This study was performed in the University of Wuhan, Wuhan, China in May 2007. Sepsis models were made by cecal ligation and puncture CLP in Sprague-Dawley rats. Forty rats were randomly divided into sham, CLP, CLP/UTI I 20u/g and CLP/UTI II 50u/g groups, with 10 rats in each. All of them were sacrificed 12 hours after CLP. The mean arterial pressure MAP, heart rate HR, the wet-to-dry lung weight ratio W/D was measured and venous blood was collected for assaying tumor necrosis factor-alpha TNF-alpha, interleukin-10 IL-10, alanine aminotransferase ALT, aspartate aminotransferase AST and lactic acid. Superoxide dismutase SOD, malondialdehyde MDA and expression of inducible nitric oxide synthase iNOS mRNA in lung and hepatic tissues were examined. Compared with the CLP group, MAP and HR in 50u/g UTI treated rats was stable p<0.01. Marked elevation levels of W/D ratio were lowered after administration of 50u/g UTI p<0.01. Treatment with 50u/g UTI prevented marked elevation in MDA, ALT, AST, TNF-alpha, lactic acid levels, expression of iNOS mRNA, and elevated IL-10 and SOD activity p<0.01. Urinary trypsin inhibitor has a protective effect against sepsis. Its action mechanisms are probably involved in the inhibition of inflammatory factor production and suppression of lipid peroxidation and iNOS mRNA expression


Subject(s)
Animals, Laboratory , Sepsis/therapy , Rats , Lipid Peroxidation , Treatment Outcome , Cytokines , Blood Pressure , Lung/injuries , Liver/injuries
16.
Chinese Journal of Cardiology ; (12): 57-61, 2008.
Article in Chinese | WPRIM | ID: wpr-299498

ABSTRACT

<p><b>OBJECTIVE</b>To explore the role of TLR4 in myocardial ischemia reperfusion injury (MI/RI) by observing the dynamic TLR4 expression changes at mRNA and protein levels early after myocardial ischemia reperfusion.</p><p><b>METHODS</b>Male SD rats were randomly divided into Sham and IR group and the rats were killed according to different reperfusion time (0, 0.5, 1, 2, 4 and 8 hours). Myocardial changes under light microscope and transmission electronic microscope were observed. TLR4 expressions at protein and mRNA levels were detected by immunohistochemistry and realtime RT-PCR respectively. Myocardial TNF-alpha was determined by ELISA.</p><p><b>RESULTS</b>(1) Myocardial injury was observed in IR but not in Sham group and histopathological and ultrastructural changes in IR group remained unchanged up to 8 hours after reperfusion. (2) Positive TLR4 protein staining was visualized in both Sham and IR groups and significantly increased and peaked at 1 hour of reperfusion in IR group. (3) Compared to Sham group, TLR4 mRNA level was upregulated in myocardium in IR group and peaked at 1 hour of reperfusion. (4) Concentration of TNF-alpha in IR group was significantly higher than that of Sham group at corresponding time points (all P < 0.05), and myocardial TLR4 mRNA level correlated positively with myocardial TNF-alpha (r = 0.728, P < 0.01).</p><p><b>CONCLUSION</b>Expression of TLR4 in myocardium during early after myocardial ischemia reperfusion was upregulated and actived TLR4 might play an important role in MI/RI through promoting myocardial TNF-alpha excretion.</p>


Subject(s)
Animals , Disease Models, Animal , Male , Myocardial Reperfusion Injury , Metabolism , Pathology , Myocardium , Pathology , Rats , Rats, Sprague-Dawley , Toll-Like Receptor 4 , Metabolism , Tumor Necrosis Factor-alpha , Metabolism
17.
Article in English | WPRIM | ID: wpr-236728

ABSTRACT

<p><b>OBJECTIVE</b>To examine whether TLR-4 has an effect on hemorrhage induced changes in lung, and to investigate the change of heme oxygenase-1 (HO-1) on acute lung injury (ALI) induced by hemorrhagic shock in mice.</p><p><b>METHODS</b>Forty-eight male mice, including C3H/HeN mice and C3H/HeJ mice, were randomly divided into sham group (n=12), hemorrhagic shock group with twelve mice in each phase. Blood pressure (BP) was monitored continuously by attaching carotid artery catheter to a strain gauge pressure transducer/ polygraph. Arterial blood samples were taken for blood gas analysis. A mouse model of non-lethal hemorrhagic shock and resuscitation was used to observe pulmonary myeloperoxidase (MPO) activity and wet/dry weight ratio (W/D). The expression of HO-1 was observed by means of RT-PCR and immunohistochemistry. IL-6 and IL-10 in lung tissue homogenate were assayed by enzyme-linked immunosorbent assay (ELISA). The pulmonary pathologic changes were observed under electron microscope and light microscope.</p><p><b>RESULTS</b>Compared with sham group, the expression of HO-1 in lung tissue was significantly higher in Hem 24 h and Hem 48 h of C3H/HeN mice (P less than 0.01). The expression of HO-1 mRNA and the levels of IL-6, IL-10 and MPO in lung tissue were markedly increased in Hem 24 h (P less than 0.01 or P less than 0.05); Compared with C3H/HeN mice, the expression of HO-1 mRNA and the levels of IL-6 and IL-10 in C3H/HeJ mice significantly decreased in Hem 24 h and Hem 48 h (P less than 0.01 or P less than 0.05), and the W/D, MPO in C3H/HeJ mice were obviously lower in Hem 24 h (P less than 0.05). The injuries of lung tissues after hemorrhagic shock have been demonstrated by histological examination with electron microscope and light microscope.</p><p><b>CONCLUSIONS</b>TLR-4 and HO-1 might modulate the balance of pro- and anti-inflammatory processes in inflammatory reaction of hemorrhagic shock-induced ALI, and the activation of Toll-like receptor might induce the transcription activity of HO-1, which may play a key role in acute lung injury.</p>


Subject(s)
Acute Disease , Animals , Heme Oxygenase-1 , Metabolism , Lung , Pathology , Male , Mice , Mice, Inbred C3H , Random Allocation , Shock, Hemorrhagic , Toll-Like Receptor 4 , Physiology
18.
Article in Chinese | WPRIM | ID: wpr-281522

ABSTRACT

<p><b>OBJECTIVE</b>To study the neuroprotective effect of hypoxic preconditioning on reperfusion injury following ischemia and its molecular mechanism.</p><p><b>METHODS</b>Forty-eight rats were randomized into 3 groups, namely the sham operated group, ischemia/reperfusion (I/R) group, and I/R following hypoxic preconditioning group (HP+I/R). In the latter two groups, the rats were subjected to middle cerebral artery occlusion (MACO) for 3 h followed by reperfusion for 24 h to induce cerebral I/R injury. The learning and memory ability of the rats 24 h after reperfusion was assessed using Y-maze test. Immunohistochemistry was performed to quantify the expressions of survivin and HSP-70 proteins in the rat brain tissues.</p><p><b>RESULTS</b>The number of survivin- and HSP-70-positive cells in the brain tissues was significantly different between HP+I/R group and IR and the sham operated groups (P<0.05), and following I/R injury, the rats in HP+I/R group showed much better performance in the Y-maze test than those in I/R group.</p><p><b>CONCLUSION</b>Hypoxic preconditioning can protect the ischemic brain against reperfusion injury, promote recovery of the learning and memory ability and neurological functions following the injury. Up-regulation of the expressions of survivin and HSP-70 proteins might be one of the molecular mechanisms for this neuroprotective effect.</p>


Subject(s)
Animals , Brain , Metabolism , Brain Ischemia , Therapeutics , HSP70 Heat-Shock Proteins , Metabolism , Infarction, Middle Cerebral Artery , Ischemic Preconditioning , Memory , Microtubule-Associated Proteins , Metabolism , Rats , Rats, Sprague-Dawley , Reperfusion Injury , Therapeutics
19.
Article in Chinese | WPRIM | ID: wpr-640592

ABSTRACT

Objective To investigate the related risk factors and neonatal outcomes in early preterm birth by analysis of 198 cases of early-to-moderate preterm birth. Methods Retrospective analysis was conducted on 198 pregnant women hospitalized during January 2004 to September 2006 with early-to-moderate preterm birth who delivered at 28 to 33+6 weeks of gestational age and their preterm infants.All of them were divided into two groups: early preterm birth group,28 to 3l+6 weeks of gestational age,n=90 for the pregnant women and n= 99 for the preterm infants;moderate preterm birth group,32 to 33+6 weeks of gestational age,n=108 for the pregnant women and n=122 for the preterm infants(fatal birth defects were excepted).The risk factors for preterm birth and neonatal outcomes were compared between the two groups. Results Various factors contributed to the occurrence of preterm birth.Systemic antenatal care and weight gain during pregnancy were significantly less found in the early preterm birth group than the moderate preterm birth group(P

20.
Article in Chinese | WPRIM | ID: wpr-685826

ABSTRACT

Objective: Clone, express and purify human recombinant calreticulin (CRT). Methods: Human CRT cDNA was amplified from total RNA of human lung cancer cell line A549 cells by RT-PCR. Then, PCR product was subcloned into prokaryotic expression vector pET-15b. After sequencing, this recombinant plasmid was transformed into E.coli. Rossetta. Recombinant CRT was expressed in host cells by IPTG induction. Resulted protein was purified by Ni-NTA resin under denature condition and dialyzed to recover its native structure. SDS-PAGE and Western blot method were used to identify the expression and purification of reconbinant CRT. Results: Human CRT cDNA was cloned from total RNA of A549 cells. CRT prokaryotic expression vector pET-15b-crt was constructed. Reconbinant CRT was induced to express in E.coli and purified by Ni-NTA affinity chromatograph. Conclusion: A method for prokaryotic expression and purification of human recombinant CRT was successfully established. This method laid a foundation for the subsequent CRT research.

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