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1.
Article de Chinois | WPRIM | ID: wpr-939682

RÉSUMÉ

OBJECTIVE@#To explore the expression of cellular apoptosis susceptibility protein (CAS) in acute myeloid leukemia (AML) and its correlation with clinical characteristics.@*METHODS@#The expression of CAS in bone marrow tissue of 54 patients with AML and 24 patients with non-hematological malignant diseases was detected by Western blot and immune-histochemical method, and compared between AML group and control group. Also the relationship of CAS expression in AML and sex, age, WBC count, Hb, platelet count, bone marrow blast cell ratio, ki-67 index, cytogenetic and molecular biological prognostic risk stratification, extramedullary infiltration and other clinical characteristics was analyzed.@*RESULTS@#Western blot showed that the expression of CAS protein in bone marrow biopsies of AML patients was significantly higher than that in control group (P<0.05). Immune-histochemical method revealed that CAS was mainly located in the cytoplasm in both AML group and control group. Among 54 AML patients, 14 patients (25.9%) showed high expression of CAS, while all the 24 patients in the control group showed low expression of CAS. The high expression rate of CAS in AML patients was significantly higher than that in the control group (P<0.05). There were statistically significant differences in prognostic risk stratification and the remission rate of the first chemotherapy between CAS high expression group and CAS low expression group in AML (P<0.05). The proportion of high risk patients and unremission patients after the first chemotherapy in CAS high expression group were significantly higher than those in CAS low expression group (57.1% vs 27.5%, 30.8% vs 7.9%), while the proportion of low risk patients and complete remission patients after the first chemotherapy were significantly lower than those in CAS low expression group (14.3% vs 37.5%, 53.8% vs 84.2%). In AML patients, the ki-67 index of bone marrow tissue in CAS high expression group was higher than that in CAS low expression group (60% vs 50%) (P<0.05).@*CONCLUSION@#CAS is localized in cytoplasm in both AML and non-hematological malignant diseases, and its expression increases in AML. CAS is related to the risk stratification of cytogenetics and molecular biology, the remission rate after the first chemotherapy and ki-67 index in AML, which suggests that CAS may be involved in the occurrence and development of AML.


Sujet(s)
Humains , Moelle osseuse/métabolisme , Protéine cellulaire de susceptibilité à l'apoptose/métabolisme , Antigène KI-67/métabolisme , Leucémie aigüe myéloïde/traitement médicamenteux , Pronostic , Induction de rémission
2.
Yao Xue Xue Bao ; (12): 907-914, 2020.
Article de Chinois | WPRIM | ID: wpr-821671

RÉSUMÉ

To study the osteoprotective effect of 1,2,3,4,6-pentyl-O-galloyl-beta-D-glucose (PGG) its anti-osteoblast apoptosis related mechanism was investigated. A model of zebrafish osteoporosis induced by prednisolone (Pred, 25 μmol·L-1) was established in vivo, and calcein staining was used to detect the effect of PGG on the bone area of ​​zebrafish. Bone marrow mesenchymal stem cells were cultured in vitro, and the number of calcified nodules was observed by alizarin red staining, and the relevant indexes of osteoblast differentiation runt-related transcription factor 2 (Runx 2), osteocalcin (OCN) mRNA level were detected by qRT-PCR. The osteoblast cell line MC3T3-E1 cells was cultured in vitro, and 400 μmol·L-1 hydrogen peroxide (H2O2) was used to intervene the injury to detect the effect of PGG on osteoblasts under oxidative stress. The effect of PGG on osteoblast activity was detected by MTT assay. The effect of PGG on apoptosis was observed by Hoechst 33342 staining. Western blot was used to detect the expression of Bcl-2, Bax, nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). DCFH-DA fluorescence staining for detection of reactive oxygen species (ROS) levels. JC-1 staining was used to detect mitochondrial membrane potential levels. The results showed that PGG could significantly increase the vertebral area of ​​the zebrafish model when compared with the model group. On the 14 th day of osteoblast differentiation, the number of calcified nodules in the PGG group was significantly increased when compared with the control group and the mRNA levels of Runx 2 and OCN were also significantly increased. In addition, under oxidative stress, PGG could increase osteoblast viability, significantly reduce the number of apoptotic cells, and increase the ratio of Bcl-2/Bax. Fluorescence staining results show that PGG decreased intracellular ROS fluorescence density and increased mitochondrial membrane potential. Western blot data showed that PGG could promote the expression of Nrf2 in the nuclear and enhance the expression of downstream protein HO-1. In conclusion, PGG could improve osteoporosis in zebrafish, and this effect may be related to the regulation of Nrf2/HO-1 signaling pathway to improve mitochondrial dysfunction, anti-oxidative stress in osteoblast apoptosis and promote bone formation. This study provides new ideas and clues for the discovery of anti-osteoporosis drugs.

3.
Article de Chinois | WPRIM | ID: wpr-664593

RÉSUMÉ

Thioredoxin-interacting protein ( TXNIP) suppresses the antioxidative function of thioredoxin ( Trx ) by combining with thioredoxin ( Trx).Therefore, it promotes the generation and accumulation of reactive oxygen species ( ROS ) , inducing endoplasmic reticulum stress and mitochondrial stress , which leads to cellular inflammation or cellular apoptosis ultimately . TXNIP-mediated oxidative stress plays a crucial role in control-ling the generation and development of some diseases , such as diabetes and its complications ( diabetic nephropathy diabetic retinopathy etc .) , atherosclerosis ischemia/reperfusion injury , cancers ( hepatocellular carcinoma , carcinoma of urinary blad-der, mammary cancer , leukemia ) etc.Here, we try to review the action and mechanism of oxidative stress mediated by TXNIP in the diseases and the progress in research .

4.
Yao Xue Xue Bao ; (12): 1404-1409, 2017.
Article de Chinois | WPRIM | ID: wpr-779741

RÉSUMÉ

To investigate the effects of cordycepin on proliferation and invasion of pancreatic cancer stem cells (Pan CSC) and its mechanisms, MTT assay was used to investigate the effect of cordycepin on proliferation of Pan CSC. Inverted microscope was used to observe the morphologic change of cells. Propidium iodide staining methods was employed to observe the cell apoptosis. Cell scratch method was used to detect the ability of migration of Pan CSC in each group. RT-PCR and Western blot were used to determine the expression of apoptosis gene and epithelial-mesenchymal transitions (EMT) gene. The growth of Pan CSC was inhibited by cordycepin in a dose-and time-dependent manner, with IC50 107.364 and 48.472 μmol·L-1 at 24 and 48 h, respectively. Moreover, the cell migration was inhibited at the same time. RT-PCR and Western blot results showed that cordycepin decreased the expression of Bcl-2 and activated pro-apoptotic gene levels such as Bax,p53, caspase-3. Furthermore, cordycepin reduced the expression of EMT genes by up-regulation of E-cadherin and down-regulation of N-cadherin. Cordycepin has the ability to inhibit Pan CSC proliferation and invasion by activating p53 pathway as well as suppressing the EMT. This study provides a new basis for inhibition of pancreatic cancer stem cells in the treatment of pancreatic cancer.

5.
Chinese Journal of Immunology ; (12): 1441-1445, 2016.
Article de Chinois | WPRIM | ID: wpr-504375

RÉSUMÉ

Objective:To investigate the mechanism of Smad7 gene modified bone marrow mesenchymal stem cells ( Smad7-BMSCs) to prevent hepatic fibrosis in vitro. Methods:Smad7-EGFP-BMSCs were established by isolating and purifying BMSCs of rats, and transfecting Ad-Smad7-EGFP. HSC-T6 were divided into Group A, Group B, Group C and Group D, which were respectively incubated with Smad7-EGFP-BMSCs, BMSCs, Smad7 plasmid and PBS for 72 hours. The level of Smad7and TGF-β1protein in the culture solution was determined by ELISA. The expression of mRNA and protein of Smad7,TGF-β1,Col Ⅰ and α-SMA in the hepatic stellate cells were respectively determined by Western blot and RT-PCR. Cellular apoptosis was determined by flow cytometry. Results:(1)The results of ELISA showed that the level of TGF-β1 protein decreased(P<0. 01) but the level of Smad7 protein increased (P<0. 01) in Group B,Group C and Group D compared with Group A;the level of TGF-β1 protein decreased(P<0. 01) but the level of Smad7 protein increased (P<0. 01) in Group D compared with Group B and Group C. (2)The results of Western blot and RT-PCR showed that the level of mRNA and protein of Smad7,TGF-β1,Col Ⅰ and α-SMA decreased(P<0. 01) but the level of mRNA and protein of Smad7 protein increased (P<0. 01) in Group B,Group C and Group D compared with Group A;the level of mRNA and protein of Smad7,TGF-β1,ColⅠandα-SMA decreased(P<0. 01) but the level of mRNA and protein of Smad7 protein increased (P<0. 01) in Group D compared with Group B and Group C. (3)The results of flow cytometry showed that the rate of cellular apoptosis de-creased(P<0. 01),but the level of Smad7 protein increased (P<0. 01) in Group B,Group C and Group D compared with Group A;the rate of cellular apoptosis decreased(P<0. 01)in Group D compared with Group B and Group C. Conclusion:Smad7-BMSCs can have the effect of anti-hepatic fibrosis by affecting TGF-β1 signal pathway and promoting cellular apoptosis in hepatic stellate cells.

6.
Yao Xue Xue Bao ; (12): 1845-2016.
Article de Chinois | WPRIM | ID: wpr-779341

RÉSUMÉ

This study was designed to investigate the mechanism of 5,2',4'-trihydroxy-6,7,5'-trimethoxy flavone nanoparticle (TTF1-NP) in the induction of apoptosis of human hepatocellular carcinoma HepG-2 cells. MTT assay, immunocytochemical staining and flow cytometry with Annexin V-FITC/PI were used to demonstrate inhibition of proliferation of HepG-2 cells and cell apoptosis. The inhibition was studied in a dose- and time-dependent manner. Western blot results showed that TTF1-NP down-regulated the signals of survivin, p-STAT3 and STAT3, but up-regulated the expression level of cleaved caspase-3. Taken together, our results showed that TTF1-NP induced HepG-2 cell apoptosis through inhibition of the STAT3 expression.

7.
Article de Chinois | WPRIM | ID: wpr-839634

RÉSUMÉ

Objective To analyze the relationship of CAS protein expression with proliferative index, apoptosis index and clinical parameters in breast cancer tissues. Methods Immunohistochemistry for CAS expression, ki-67 (proliferative index) and TUNEL (apoptosis index) were examined in 20 usual ductal hyperplasia (UDH), 20 atypical ductal hyperplasia (ADH), 10 ductal carcinoma in situ (DCIS), 53 invasive ductal carcinoma(IDC) and 14 normal breast tissues. Results CAS expression increased in order in the normal breast tissues, UDH, ADH, DCIS and IDC, with the positive rates of CAS protein being 14.3% (2/14), 25.0% (0/20), 40.0% (8/20), 60.0% (6/10), and 75. 5% (40/53), respectively {χ2 = 29. 382, P = 0. 000). CAS protein expression was correlated with histological grade, mitotic activity, and lymph node metastasis of IDC (P<0. 01, P<0. 05), and not with patient age, tumor volume or grade. CAS protein expression was positively correlated with ki-67 index (r = 0. 439, P = 0. 003), and not with the apoptosis index (r=0. 248, P = 0. 083). Conclusion CAS protein expression is associated with cell proliferation index in breast cancer tissues.

8.
Journal of Chinese Physician ; (12): 584-586,591, 2011.
Article de Chinois | WPRIM | ID: wpr-597858

RÉSUMÉ

Objective To analyze the apoptosis-related spots like protein (ASC) and caspase -1 in HCC and adjacent noncancerous tissues and its clinical significance. Methods ASC and Caspase 1 were determined with immunohistochemistry LSAB method and the various indexes were analyzed in 30 cases of liver cancer and adjacent tissues which did not receive preoperative radiotherapy and chemotherapy. Results The positive rate of ASC in HCC and adjacent noncancerous tissues was 10.00% (3/30) and 73.33% (22/30). Positive rate between the groups had significant difference (P<0.01). Caspase-1 positive rate had significantly difference between the groups (23.33% (7/30) vs. 66.67% (20/30), P<0.01). ASC and the two indicators of Caspase-1 expression in cancer tissues were low, but no significant correlations between the two groups (P>0.05). ASC and the two indicators of Caspase-1 expression in adjacent tissue were significantly increased, and it showed a very significant correlation (r=0.722,P<0.01). Conclusions ASC, Caspase-1 expression in liver cancer were low expressed, and significantly lower than the adjacent tissues, which suggested that ASC and Caspase-1 were closely related to the occurrence and development of the apoptosis of liver cancer, and it might be used as diagnosis and treatment of targets in liver cancer.

9.
Journal of Acupuncture and Tuina Science ; (6): 328-332,封二, 2006.
Article de Chinois | WPRIM | ID: wpr-601817

RÉSUMÉ

To investigate the mechanism of moxibustion in regulating cellular apoptosis in rat's precancerous lesion of primary hepatocellular carcinoma (HCC). Methods:Seventy-four rats were randomly allocated to normal group,model group and moxibustion group,and the diethylic nitrosamine (DEN) was used to establish HCC model. Moxibustion with moxa cone which is as big as a grain of wheat was performed on acupoint Zusanli (ST 36),3 cones for each acupoint and 0.5 mg for each cone,the treatment was given once a day,totally 16 weeks. Then the changes in the body weight,liver weight and thymus weight,a morphological change in the liver tissue and changes in γ-GT and GST were observed;Immunohistochemical staining method was adopted to observe the tendency of changes in relevant apoptosis genes such as C-myc,N-ras and mutant type P53,and the influence of moxibustion on cell cycle modulation genes such as cyclinD1,CDK4 and p16. Results:Moxibustion could reduce the activities of γ-GT and GST in the blood,obviously decrease the protein expression of relevant apoptosis genes such as C-myc,N-ras and mutant type P53 and markedly inhibit the over-expression of relevant cell cycle modulation genes such as cyclinD1 and CDK4 and the mutation of cell cycle modulation gene p16. Conclusion:Moxibustion might play a certain role in relieving HCC precancerous lesion and its action mechanism might be related to the regulation on partial apoptosis genes.

10.
Article de Chinois | WPRIM | ID: wpr-588747

RÉSUMÉ

Objective To observe the effects of electrical stimulation of paraventricular nucleus(PVN) on gastric mucosal cellular apoptosis,proliferation,and expression of BCL-2,BAX induced by gastric ischemia-reperfusion(GI-R) and the potential mechanisms of protection of PVN on GI-R injury.Methods After electrical stimulation of PVN,the experimental model of GI-R were established by clamping the celiac artery for 30 min and then reperfusing the artery for 30 min,1 h,3 h,or 6 h respectively.We used immunohistochemistry to detect the gastric mucosal cells apoptosis,proliferation and the expression of BCL-2,BAX.Results Compared with GI-R group,the electrical stimulation of PVN markedly decreased gastric mucosal cellular apoptosis,increased the proliferation,and promoted the protein expression of BCL-2,but markedly inhibited the protein expression of BAX at 30 min,1 h,3 h after reperfusion respectively.Conclusion The protective effect of PVN on GI-R injury is associated with up-regulation of expression of BCL-2 and down-regulation expression of BAX,and so inhibited gastric mucosal cellular apoptosis and promoted proliferation.

11.
Article de Chinois | WPRIM | ID: wpr-593085

RÉSUMÉ

Objective To observe effects of Apelin-13 microinjection into the hypothalamic paraventricular nucleus(PVN) on gastric ischemia-reperfusion(GI-R) injury in rats.Methods After Apelin-13 injection into PVN,the experimental model of GI-R was established by clamping the celiac artery for 30 min and then reperfused the artery for 1 h.We used immunohistochemistry to detect the gastric mucosal cells apoptosis,proliferation and the expression of BCL-2,BAX.Results(1)Apelin-13 microinjection into the PVN aggravated GI-R injury in an dose-dependent manner with dosages as 0.2,1.0 or 5.0 ?g,respectively.(2)Compared with GI-R group,Apelin-13 microinjection into PVN markedly increased gastric mucosal cellular apoptosis,decreased the proliferation and promoted protein expression of BAX,but obviously inhibited the protein expression of BCL-2.Conclusion Apelin-13 microinjection into the PVN may aggravate GI-R injury by promoting gastric mucosal cellular apoptosis and inhibiting proliferation.

12.
Article de Chinois | WPRIM | ID: wpr-680545

RÉSUMÉ

Objective To observe the effect of allogenic bone marrow mononuclear cells(BM-MNCs) transplantation on myocardial apoptosis after acute myocardial infarction(AMI) in rats.Methods 40 Wistar rats were randomly divided into control group(n=20) and transplantation group(n=20).Myocardium around the infarcted left ventricular area of the rats in transplantation group were injected with BM-MNCs suspension beneath the epicardium.Myocardium the area of control group was injected with culture solution.Results After 4 weeks of the operation,the myocardial apoptosis index,the TNF-? content and the PDCD5 mRNA of transplantation group were all notably less than those of control group(P

13.
Article de Chinois | WPRIM | ID: wpr-532802

RÉSUMÉ

Objective To study the effect of immunomodulation on improvement of immune function and prognosis in sepsis in rats,and its mechanism.Methods Experimental part: cecal ligation-perforation(CLP) models were divided into three groups including sham group(n=20),control group(n=20) and experimental group(n=20).Control group only used antibiotic and experimental group used antibiotic plus immunomodulation.Blood collections were made after CLP model at 3,12,48 and 72hr.Lymphocyte counting,CD4+,CD8+ T lymphocyte and CD4/CD8 ratio were checked.The apoptosis of lymphocyte in thymus and spleen and survival rate were checked.Clinical part: Prospective analysis of seventy patients who conformed to the sepsis standard.They were divided into two groups randomly.One was control group with regular therapy,and the therapy group with ulinastatin plus thymosin-?1 for 7days.The immune index before and after therapy at 0,1d,3d,and 7d was observed,including the clinical changes and survival data.Results Experimental part: Lymphocytes,CD4+ T lymphocytes and CD4/CD8 ratio in experimental group increased more significantly than in control group(P

14.
Article de Chinois | WPRIM | ID: wpr-562804

RÉSUMÉ

Objective To study the activation effect of HBV RNase H protein on the transcription of cellular apoptosis susceptibility gene (CAS). Methods The promoter of DNA sequence of CAS gene was identified in GenBank by bioinformatics and amplified from HepG2 genome by PCR using sense (5′-GGTACCCGATTACATGTTGTACATGAAGG-3′) and antisence (5′-CTCGAGGCTGAGTTCCATTGCTATAG-3′) primers. As these primers contained Kpn I and Xho I recognition sites on their respective 5′-ends, the amplified DNA fragments were tested by sequencing and then subcloned into Kpn I/Xho I sites of pCAT3-Basic reporter vector by routine molecular biological methods. The reconstructed plasmid named pCAT3-CASp was identified by enzyme digestion of Kpn I/Xho I, in which the expression of chloramphenical acetyltransferase (CAT) was under the control of the promoter of CAS. The HepG2 cells were transfected by pCAT3-CASp, and then co-transfected by pCAT3-CASp and pcDNA3.1(-)-RH plasmids. At the same time, the empty pCAT3-basic and pCAT3-TXNRD1p were transfected (self-contructed by the authors) as controls. After 24h culturing, cells were collected and the expression of CAT activity was detected by ELISA according to the manufacturer′s protocol. Results The optical density of expression of CAT of pCAT3-CASp was 0.043 by ELISA, in contrast, the optical density of expression of pCAT3-Basic was 0.024. The expression of CAT in co-transfection of pCAT3-CASp and pcDNA3.1(-)-RH(0.065) was 1.5 times as higher as pCAT3-CASp plasmid (0.043), and 2.7 times as higher as pCAT3-Basic. Conclusions The CAS gene promoter identified in present study has transcription activity and HBV RNase H protein may activate the expression of CAS gene.

15.
Article de Chinois | WPRIM | ID: wpr-547252

RÉSUMÉ

Objective To investigate the apoptosis of cultured normal human hepatocyte HL-7702 cells induced by glycodeoxycholate(GCDC)and to explore its possible mechanism.Methods HL-7702 cells were incubated with various concentrations(100,150,200 and 250 ?mol/L)of GCDC.The changes of cellular morphology were observed under optical microscope.The apoptosis rate of HL-7702 was determined by Annexin V-FITC/PI double staining.The changes of HL-7702 cell intracelluar \[Ca2+\]i were determined with Fluo-3/AM load technique.The mRNA expression levels of Bcl-2/Bax in HL-7702 cells were analyzed by RT-PCR.Results Typical apoptotic morphological changes were observed after HL-7702 cells had been treated with 150 ?mol/L GCDC for 24 h;HL-7702 cells could be induced to undergo apoptosis in a concentration-dependent manner after 100,150,200,and 250 ?mol/L GCDC treatment for 24 hours.The apoptosis rates were(13.16?2.9)%,(20.3?3.0)%,(25.02?2.1)% and(45.02?3.5)%,which were markedly higher(P

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