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1.
Article in English | WPRIM | ID: wpr-879307

ABSTRACT

Objective To assess the effects of vascular endothelial growth factor-C (VEGF-C)/vascular endothelial growth factor receptor-3 (VEGFR-3) signaling on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in human osteosarcoma MG63 cells and the subsequent impact on the proliferation of human umbilical vein endothelial cells (HUVECs). MethodsMG63 cells were treated with VEGF-C alone (VEGF-C group), VEGF-C + iNOS inhibitor aminoguanidine (AG; AG group), and VEGF-C + VEGFR-3 inhibitor MAZ51 (MAZ51 group); untreated MG63 cells were used as controls. NO production was evaluated by a colorimetric method involving nitrate reductase. Meanwhile, mRNA and protein levels of iNOS were examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. To explore the effect of VEGF-C/VEGFR-3/iNOS signaling of MG63 cells on proliferation of HUVECs, we set up six groups: HUVECs, HUVECs+MG63, HUVECs+VEGF-C, HUVECs+MG63+VEGF-C, HUVECs+MG63+VEGF-C+AG, and HUVECs+MG63+VEGF-C+MAZ51 groups. The proliferation of HUVEC cells was assessed by Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay, and proliferating cell nuclear antigen (PCNA) expression quantitation. ResultsVEGF-C treatment enhanced iNOS expression at both gene and protein levels (mRNA: LSD-

2.
Article in Chinese | WPRIM | ID: wpr-879391

ABSTRACT

OBJECTIVE@#To investigate relationship between cold pain of knee joint and subchondral bone marrow edema (BME).@*METHODS@#From May 2018 to August 2019, 92 patients with knee osteoarthritis (KOA) associated with cold pain of knee were admitted, all patients were underwent MRI examination. The patients were divided into observation group (47 patients with BME) and control group(45 patients without BME). In observation group, there were 6 males and 41 females aged from 36 to 87 years old with an average of (63.2±12.3) years old. In control group, there were 10 males and 35 females, aged from 48 to 84 years old with an average of (62.7±8.3) years old. All patientswere treated with drugs. The degree of joint degeneration was evaluated by Kellgren-Lawrence (K-L) grading. Degree of cold pain of knee was evaluated by knee cold pain score, and degree of BME was evaluated according to WORMS. The correlation between cold pain of knee and K-L grading and BME was analyzed.@*RESULTS@#Score of cold pain in observation group (15.55±7.68) was higher than that of control group (9.42± 5.50), which had significant difference (@*CONCLUSION@#The cold pain of KOA patients is not related to K-L grading, but corelate with BME grading. The Cold pain of knee was more pronounced in KOA patients with BME, and the severity of BME is often related to degree of cold pain. It seemed to be a tendency:the more serious BME, the heavier coldpain.


Subject(s)
Adult , Aged , Aged, 80 and over , Bone Marrow , Edema , Female , Humans , Knee Joint/diagnostic imaging , Magnetic Resonance Imaging , Male , Middle Aged , Osteoarthritis, Knee/diagnostic imaging , Pain/etiology
3.
Article in Chinese | WPRIM | ID: wpr-793052

ABSTRACT

As an important branch of artificial intelligence,the emerging medical artificial intelligence(MAI)is facing many ethical issues.MAI may offer the optimal diagnosis and treatment for patients but may also bring adverse effects on society and human beings.This article discusses the ethical problems caused by MAI and elucidates its development in a direction that meets ethical principles and requirements.

4.
Acta Pharmaceutica Sinica ; (12): 2679-2687, 2020.
Article in Chinese | WPRIM | ID: wpr-837530

ABSTRACT

Peptidyl-prolyl cis-trans isomerase Pin1 is over-expressed in prostate cancer cells and the level of expression correlates with the malignancy grade and prognosis in patients. In this work, twenty-one 2-(1H-benzimidazol-2-ylthio) acetic acid derivatives were designed and prepared with the aid of the crystal structure of Pin1 and our previous work. The chemical structures of the target compounds were confirmed by 1H NMR, 13C NMR, ESI-MS and IR. The inhibitory activity of compounds 6a-6i and 13a-13i against Pin1 were determined using a protease-coupled assay. The results indicated that twenty compounds were significantly superior to the positive control drug Juglone, and 6g, 6h and 13i exhibited the most potent Pin1 inhibitory activity, with IC50 values at the sub-micromolar level. The in vitro anti-proliferative activities of these analogs were evaluated by the MTT assay and several showed a moderate effect in human prostate cancer PC-3 cells. Molecular docking studies demonstrated that both the benzimidazole skeleton and the thioacetic acid fragment were indispensable for the compounds to interact with key residues in the catalytic domain of Pin1.

5.
Article in Chinese | WPRIM | ID: wpr-826329

ABSTRACT

Cervical cancer is the second most common malignant tumor in women worldwide.The burden of cervical cancer is particularly heavy in less developed countries as the malignancy brings huge pain to the patients and their family members and causes huge losses to social development and global health.However,cervical cancer is a preventable and curable disease.While screening and human papillomavirus vaccination in developed countries have remarkably lowered the incidence and mortality of cervical cancer,there is still a far way to go to achieve the prevention and treatment of this disease.The multidisciplinary prevention and control programs slightly differ in different countries due to diverse economic and health conditions.The general principle is to vaccinate the young females and to implement a comprehensive strategy including human papillomavirus vaccine vaccination,screening,early diagnosis,and early treatment in adults.


Subject(s)
Early Detection of Cancer , Female , Humans , Papillomavirus Infections , Papillomavirus Vaccines , Uterine Cervical Neoplasms , Vaccination
6.
Journal of Medical Postgraduates ; (12): 598-603, 2020.
Article in Chinese | WPRIM | ID: wpr-821835

ABSTRACT

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

7.
Journal of Medical Postgraduates ; (12): 598-603, 2020.
Article in Chinese | WPRIM | ID: wpr-821815

ABSTRACT

ObjectiveThere are few studies on whether the occurrence of anti-tuberculosis drug-induced liver injury (ADIH) is associated with the polymorphism of CYP2E gene and methylation level. This study aims to CYP2E1 gene polymorphism and the relationship between the methylation level of the promoter region and ADIH in Mongolian tuberculosis (TB) patients.Methods A total of 135 Mongolian TB patients who received standardized treatment at the Tuberculosis Research Institute of Tongliao City, Inner Mongolia from November 2015 to June 2018 were selected. According to the ADIH criteria, TB patients with liver injury were selected as the ADIH group (n=45), and TB patients without liver injury were matched as the control group based on a ratio of 1∶2 (n=90). DNA extraction and polymerase chain reaction (PCR) were performed to amplify the CYP2E1 gene to determine the CYP2E1 rs2031920 genotype, and to analyze the CYP2E1 gene polymorphism and relationship between ADIH and promoter methylation level.Results There were no significant differences in the distribution of CYP2E1 rs2031920 genotype, C1 and C2 gene frequencies between the ADIH group and the control group (P>0.05). The overall methylation level in the promoter region of CYP2E1 gene in ADIH group (0.711±0.085) was significantly lower than that of the control group (0.759±0.062). Results of Logistic regression showed that the overall methylation level in the promoter region of CYP2E1 gene was the influencing factor for the occurrence of ADIH (P<0.005). For each 0.1 unit increase of methylation level, the risk of ADIH occurrence reduced by 0.388 times, and the OR (95% CI) value was 0.388 (between 0.204 and 0.739).Conclusion The overall methylation level in the promoter region of CYP2E1 gene was reduced in Mongolian ADIH patients, but the polymorphism of CYP2E1 gene was not related to the occurrence of ADIH. These results suggested that CYP2E1 methylation could be applied to the prevention and treatment of ADIH in patients with tuberculosis.

8.
China Pharmacy ; (12): 1097-1102, 2020.
Article in Chinese | WPRIM | ID: wpr-821500

ABSTRACT

OBJECTIVE:To study the mechanism of enhancement effects of Astragalus complanatus polysaccharides(ACP) on the proliferation of meniscal fibrochondrocytes cells in rabbits. METHODS :The meniscal fibrochondrocytes cells were isolated from 1-month-old New Zealand white rabbits. The meniscal fibrochondrocytes cells were divided into normal control group (PBS), positive control group (glucosamine sulfate ,10 mg/mL)and ACP high-dose,medium-dose and low-dose groups (40,20,10 mg/mL). The morphology of meniscal fibrochondrocytes cells were observed under microscope. Cell proliferation rate was detected by MTT assay. Cell cycle was observed with flow cytometry. ELISA assay was used to detect relative expression of medium collagen type Ⅱ(Col Ⅱ)and alkaline phosphatase protein (ALP)in meniscal fibrochondrocytes cells. RT-qPCR and Western blotting assay were adopted to detect mRNA and protein expression of transforming growth factor β 1 (TGF-β 1) and bone morphogenetic protein 2(BMP-2). RESULTS :After cultured for 72 h,meniscal fibrochondrocytes cells were fused into a single layer,and most of them were slender type in appearance. Compared with normal control group ,the proliferation rate of meniscal fibrochondrocytes cells and the percentage of cells at G 1/G0 phase were decreased significantly in positive control group and ACP high-dose,medium-dose and low-dose groups (P<0.05);the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP,mRNA and protein expression of TGF-β1 and BMP- 2 were increased significantly (P<0.05). Compared with positive control group,inhibitory rate of meniscal fibrochondrocytes cells proliferation and the percentage of cells at G 1/G0 phase were decreased significantly in ACP high-dose group (P<0.05),while the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP , mRNA and protein expression of TGF-β1 and BMP- 2 were increased significantly (P<0.05). The inhibitory proliferation rate of meniscal fibrochondrocytes cells and the percentage of cells at G 1/G0 phase were increased significantly in ACP low-dose group (P< 0.05),while the percentage of cells at S phase ,protein expression of Col Ⅱ and ALP ,mRNA and protein expression of TGF-β1 and BMP- 2 were decreased significantly (P<0.05). There was no statistical significance in above indexes of ACP medium-dose group. CONCLUSIONS :ACP can promote the proliferation of meniscal fibrochondrocytes cells ,reduce the percentage of cells at G1/G0 phase,promote cell transformation to S phase ;the mechanism of which may be related to up-regulating TGF-β1,BMP-2 mRNA and protein expression ,promoting Col Ⅱ and ALP protein expression enhancement.

9.
Journal of Medical Postgraduates ; (12): 613-618, 2019.
Article in Chinese | WPRIM | ID: wpr-818290

ABSTRACT

Objective No study has been reported on the association between the abnormal methylation of drug metabolic enzymes and anti-tuberculosis drug-induced liver injury (ATLI). This article aimed to investigate the relationship of ATLI with the methylation of the CpG islands in the promoter regions of cytochrome P450 2E1 (CYP2E1) and glutathione s-transferase M1 (GSTM1) in Chinese Mongolian patients with tuberculosis (TB). Methods This retrospective study included 93 cases of TB diagnosed and treated in the TB prevention and treatment institutions of Tongliao, Inner Mongolia, between September 2016 and December 2017, which were divided into an ATLI (n = 31) and a non-ATLI group (n = 62), the former with and the latter without ATLI within 6 months after anti-TB medication. We compared the methylation levels of the CYP2E1 and GSTM1 genes between the two groups of patients and analyzed the risk factors of ATLI. Results In comparison with the non-ATLI controls, the patients of the ATLI group showed significantly lower methylation levels in the promoter regions of CYP2E1 (0.759 ± 0.066 vs 0.694 ± 0.091, P < 0.05) and GSTM1 (0.207 ± 0.093 vs 0.187 ± 0.092, P < 0.05). Multivariate logistic regression analysis revealed that the main risk factors of ATLI included alcohol consumption (OR = 5.329, 95% CI: 1.442-19.697, P < 0.05) and methylation in the CYP2E1 promoter region (OR = 0.312, 95% CI: 0.165-0.591, P < 0.05) in the TB patients. Conclusion ATLI is associated with the methylation level in the promoter region of the CYP2E1 gene in Chinese Mongolian patients with tuberculosis, indicating that the methylation of CYP2E1 could be used as a biomarker in the prevention and control of ATLI.

10.
Article in Chinese | WPRIM | ID: wpr-771506

ABSTRACT

In this study,transcriptomics technique was used to investigate the mechanism of action of Aconiti Lateralis Radix Praeparata on acute heart failure rats induced by propafenone hydrochloride.First,rats were randomly divided into normal group,model group and administration group(1.25,2.5,5 g·kg-1).A rat with acute heart failure was constructed by intravenous femoral administration of proparone hydrochloride.The changes of heart rate,+dp/dtmaxand-dp/dtmaxat 5,10,20,30 and 60 min were recorded.Then another group of rats were given the same drug delivery method.In another group of animals,serum TNF-α could be determined by ELISA with the same dosage method.High-throughput sequencing technology was used to detect all gene expression differences in cardiac tissue samples of rats with acute heart failure.Through functional annotation and enrichment analysis,gene expression signaling pathways of rats with acute heart failure and rats with post-administration heart failure were screened out.The results showed that heart rate and LV+dp/dtmaxand LV-dp/dtmaxwere significantly decreased in the model group(P<0.05),while heart rate and LV+dp/dtmax and LV-dp/dtmaxwere significantly increased in the drug group(P<0.05,P<0.01).Moreover,ANP,BNP and TNF-α in acute heart failure rats was significantly decreased in high-dose aconite decoction group(P<0.05).Transcriptomics analysis showed that the mechanism of action was mainly related to activation of PI3 K-AKT signaling pathway and Jak-STAT pathway.Compared with the model group,aconite decoction up-regulated the expression of phosphatidylinostol 3-kinase(PI3 K),lysophosphatidic acid(LAP3),Bcl-3 and STAT genes,and down-regulated the expression of integrin(ITGA),nuclear orphan receptor(Nur77) genes.It could be concluded that the mechanism of aconite in treating acute heart failure rats may be related to the regulation of the PI3 k-Akt/Jak-STAT pathway.


Subject(s)
Aconitum , Chemistry , Animals , Drugs, Chinese Herbal , Pharmacology , Heart , Heart Failure , Drug Therapy , Metabolism , Myocardium , Metabolism , Random Allocation , Rats , Signal Transduction , Transcriptome
11.
Article in Chinese | WPRIM | ID: wpr-278688

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the value of flow cytometry (FCM) detection in prognostic evaluation of minimal residual disease (MRD) of acute myeloid leukemia (AML) patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>Eighty-two cases of AML (except M3) after allo-HSCT who accord to enrolled condition (no MRD positive after allo-HSCT confirmed by regular flow cytometry detection and followed-up for 2 years) from April 2012 to September 2016 in our department were selected. Among 82 cases males were 50 and females were 32 with average age of 27.27 (2-57) years old. According to FAB classification, 2 cases were classified as M0/M1, 51 cases as M2, 24 cases as M4/M5 and 5 cases as M6. The antibody panels were selected accordingly to the initial leukemia associated immunophentype(LAIP) of patients.</p><p><b>RESULTS</b>Twenty patients (24.39%) were identified as MRD(0.10%-4.91%, mean 1.64%) in 82 AML patients after allo-HSCT (all the patients were in complete remission phase based on bone marrow morphology). During follow-up, 16 cases relapsed (relapse rate 80%)in 20 MRDcases, including 1 case with extramedullary relapse; 4 out of 62 MRDcases relapsed (relapse rate 6.45%)in bone marrow, and 2 cases extramedullary relapsed. The average survival time of leukemia- free survival (LFS) in group MRDwas 15.19 ± 3.99 months, median LFS was 10± 3.84 months. The average LFS time was 53.50 ± 1.69 months in MRDgroup (P<0.001). The average overall survival (OS) of the MRDgroup was 22.52 ± 5.72 months, the median OS was 18 ± 3.27 months; the average OS time was 42.86 ± 2.83 months in MRDgroup(P=0.008).</p><p><b>CONCLUSION</b>For the patients with morphologically complete remission after allo-SCT, the FCM regular monitoring of bone marrow MRD closely relates with its relapse rate, LFS and OS. Compared with the MRDgroup, the relapse rate of MRDgroup is significantly increases, and the LFS and OS significantly decreases.</p>

12.
Acta Physiologica Sinica ; (6): 565-570, 2018.
Article in Chinese | WPRIM | ID: wpr-687794

ABSTRACT

Son of sevenless homolog 1 (SOS1) protein is a ubiquitously expressed adapter. As a key protein in intracellular signaling, SOS1 plays an important role in many signal transduction pathways, such as Ras and Rac signaling pathways. The abnormal expression or mutation of SOS1 is closely related to clinical diseases. In this article, we review research progress on SOS1 functions and its roles in physiology and pathophysiology.

13.
Chinese Journal of Pathophysiology ; (12): 1551-1557, 2017.
Article in Chinese | WPRIM | ID: wpr-660669

ABSTRACT

AIM:To investigate the inhibitory effect of ethanol extract of propolis (EEP) on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell apoptosis and the underlying molecular mechanisms.METHODS:Human umbilical vein endothelial cells (HUVECs) were pretreated with EEP (7.5,15 and 30 mg/L) or 4-phenylbutyric acid (PBA,4 mmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin (TM,4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and Annexin V-FITC/PI double staining,respectively.The activities of lactic dehydrogenase (LDH) in the medium and caspase-3 in the HUVECs were measured.The protein and mRNA levels of C/EBP homologous protein (CHOP),a proapoptotic molecule under endoplasmic reticulum stress (ERS),and its downstream Bcl-2 were examined by Western blot and real-time PCR,respectively.RESULTS:Like PBA (an ERS inhibitor),EEP protected HUVECs from ox-LDL-induced injury in a dose-dependent manner,as assessed by the increased cell viability and the decreased LDH release,apoptotic rate and caspase-3 activation.The decrease in cell viability and the increases in LDH release,apoptotic rate and caspase-3 activation induced by TM,an ERS inducer,were also attenuated by EEP.Moreover,EEP suppressed ox-LDL-induced CHOP upregulation and Bcl-2 downregulation,and this effect was similar to that of PBA.Similarly,EEP significantly suppressed TM-induced CHOP upregulation both at the protein and mRNA levels.CONCLUSION:EEP may protect HUVECs from ox-LDL-induced apoptosis,and the mechanism is at least partially involved in suppressing CHOP-mediated ERS-associated apoptotic pathway.

14.
Chinese Journal of Pathophysiology ; (12): 1551-1557, 2017.
Article in Chinese | WPRIM | ID: wpr-662754

ABSTRACT

AIM:To investigate the inhibitory effect of ethanol extract of propolis (EEP) on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell apoptosis and the underlying molecular mechanisms.METHODS:Human umbilical vein endothelial cells (HUVECs) were pretreated with EEP (7.5,15 and 30 mg/L) or 4-phenylbutyric acid (PBA,4 mmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin (TM,4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and Annexin V-FITC/PI double staining,respectively.The activities of lactic dehydrogenase (LDH) in the medium and caspase-3 in the HUVECs were measured.The protein and mRNA levels of C/EBP homologous protein (CHOP),a proapoptotic molecule under endoplasmic reticulum stress (ERS),and its downstream Bcl-2 were examined by Western blot and real-time PCR,respectively.RESULTS:Like PBA (an ERS inhibitor),EEP protected HUVECs from ox-LDL-induced injury in a dose-dependent manner,as assessed by the increased cell viability and the decreased LDH release,apoptotic rate and caspase-3 activation.The decrease in cell viability and the increases in LDH release,apoptotic rate and caspase-3 activation induced by TM,an ERS inducer,were also attenuated by EEP.Moreover,EEP suppressed ox-LDL-induced CHOP upregulation and Bcl-2 downregulation,and this effect was similar to that of PBA.Similarly,EEP significantly suppressed TM-induced CHOP upregulation both at the protein and mRNA levels.CONCLUSION:EEP may protect HUVECs from ox-LDL-induced apoptosis,and the mechanism is at least partially involved in suppressing CHOP-mediated ERS-associated apoptotic pathway.

15.
Acta Physiologica Sinica ; (6): 733-739, 2016.
Article in Chinese | WPRIM | ID: wpr-331609

ABSTRACT

The purpose of the present study was to investigate the effect of advanced glycated albumin (AGE-alb) on the activation of caspase-12, a key molecule in endoplasmic reticulum stress (ERS)-associated apoptotic pathway, and to elucidate the underlying molecular mechanisms of macrophage apoptosis. RAW264.7 macrophages were treated with AGE-alb (2, 4 and 6 g/L), control albumin (C-alb, 4 g/L), tunicamycin (TM, 4 mg/L), or pretreated with 4-phenylbutyric acid (PBA, 5 mmol/L) for 1 h and then treated with AGE-alb (4 g/L). After incubation for 24 h, the cell viability and apoptosis were determined by using MTT assay and TUNEL detection kit, respectively. Lactate dehydrogenase (LDH) activity in media was determined by using an assay kit. The protein levels of caspase-12 were examined by Western blot analysis. The results showed that like TM (an ERS inducer), incubation with AGE-alb led to significant decrease in viability and increase in LDH activity in media and apoptotic rate in a dose-dependent manner. In addition, AGE-alb induced activation of caspase-12 especially at the concentration of 4 and 6 g/L (P < 0.01), which was similar to TM. However, PBA (an ERS inhibitor) protected RAW264.7 macrophages from AGE-alb-induced decrease in viability and increases in LDH activity and apoptosis. Moreover, PBA also inhibited the caspase-12 activation induced by AGE-alb (P < 0.05). These results suggest that AGE-alb may induce apoptosis in RAW 264.7 macrophages, and the mechanism may be related to the activation of ERS-associated apoptotic pathway mediated by caspase-12.


Subject(s)
Animals , Apoptosis , Caspase 12 , Cell Line, Tumor , Cell Survival , Endoplasmic Reticulum Stress , Macrophages , Mice , Phenylbutyrates , Serum Albumin , Tunicamycin
16.
Chinese Journal of Pathophysiology ; (12): 2192-2198, 2016.
Article in Chinese | WPRIM | ID: wpr-506578

ABSTRACT

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .

17.
Article in Chinese | WPRIM | ID: wpr-286384

ABSTRACT

<p><b>OBJECTIVE</b>To observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.</p><p><b>METHODS</b>NB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.</p><p><b>RESULTS</b>MAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>MAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.</p>


Subject(s)
Alkaloids , Antineoplastic Agents , Cell Differentiation , Cell Line, Tumor , Down-Regulation , Humans , Leukemia, Promyelocytic, Acute , Metabolism , Phospholipid Transfer Proteins , Metabolism , Quinolizines , RNA, Messenger , Signal Transduction , Tretinoin , Tumor Cells, Cultured , Up-Regulation
18.
Article in Chinese | WPRIM | ID: wpr-232196

ABSTRACT

<p><b>OBJECTIVE</b>To screen for potential mutations in an ethnic Han Chinese family from Shanxi with hereditary multiple exostoses.</p><p><b>METHODS</b>Polymerase chain reaction and DNA sequencing were used to screen potential mutations in EXT1 and EXT2 genes.</p><p><b>RESULTS</b>For EXT1 gene, two synonymous mutations (P477P and E587E), three intronic mutations (c.1537 -48A>G, c.1721 +203A>G and c.1722 -103C>G) were detected. For EXT2 gene, five intronic mutations (c.-29 -148A>T, c.1080 -18T>A, c.1336 -93C>T, c.1526 -166C>T, and c.1526 -195C>T) were identified. Among these, EXT1 P477P, EXT1 E587E and EXT2 c.1080 -18T>A are polymorphisms listed by Multiple Osteochondroma Mutation Database, whilst the other 7 sites have not been reported.</p><p><b>CONCLUSION</b>No mutations have been found among all exons of the EXT1 and EXT2 genes in this family. Linkage analysis is necessary for identifying the cause of this disease.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Asians , Genetics , Base Sequence , Child , Child, Preschool , China , Exons , Exostoses, Multiple Hereditary , Diagnosis , Genetics , Female , Genotype , Humans , Introns , Male , Middle Aged , Mutation , N-Acetylglucosaminyltransferases , Genetics , Young Adult
19.
Chinese Medical Journal ; (24): 2912-2917, 2013.
Article in English | WPRIM | ID: wpr-263558

ABSTRACT

<p><b>BACKGROUND</b>Previous studies have shown that brain functional activity in the resting state is impaired in Alzheimer's disease (AD) patients. However, alterations in intrinsic brain activity patterns in mild cognitive impairment (MCI) patients are poorly understood. This study aimed to explore the differences in regional intrinsic activities throughout the whole brain between aMCI patients and controls.</p><p><b>METHODS</b>In the present study, resting-state functional magnetic resonance imaging (fMRI) was performed on 18 amnestic MCI (aMCI) patients, 18 mild AD patients and 20 healthy elderly subjects. And amplitude of low-frequency fluctuation (ALFF) method was used.</p><p><b>RESULTS</b>Compared with healthy elderly subjects, aMCI patients showed decreased ALFF in the right hippocampus and parahippocampal cortex, left lateral temporal cortex, and right ventral medial prefrontal cortex (vMPFC) and increased ALFF in the left temporal-parietal junction (TPJ) and inferior parietal lobule (IPL). Mild AD patients showed decreased ALFF in the left TPJ, posterior IPL (pIPL), and dorsolateral prefrontal cortex compared with aMCI patients. Mild AD patients also had decreased ALFF in the right posterior cingulate cortex, right vMPFC and bilateral dorsal MPFC (dMPFC) compared with healthy elderly subjects.</p><p><b>CONCLUSIONS</b>Decreased intrinsic activities in brain regions closely related to episodic memory were found in aMCI and AD patients. Increased TPJ and IPL activity may indicate compensatory mechanisms for loss of memory function in aMCI patients. These findings suggest that the fMRI based on ALFF analysis may provide a useful tool in the study of aMCI patients.</p>


Subject(s)
Aged , Alzheimer Disease , Amnesia , Brain , Cognitive Dysfunction , Female , Humans , Magnetic Resonance Imaging , Male
20.
Chinese Medical Journal ; (24): 858-862, 2012.
Article in English | WPRIM | ID: wpr-269336

ABSTRACT

<p><b>BACKGROUND</b>Previous studies have shown that the functional brain activity in the resting state is impaired in Alzheimer's disease (AD) patients. However, most studies focused on the relationship between different brain areas, rather than the amplitude or strength of the regional brain activity. The purpose of this study was to explore the functional brain changes in AD patients by measuring the amplitude of the blood oxygenation level dependent (BOLD) functional MRI (fMRI) signals.</p><p><b>METHODS</b>Twenty mild AD patients and twenty healthy elderly subjects participated in the fMRI scan. The amplitude of low frequency fluctuation (ALFF) was calculated using REST software.</p><p><b>RESULTS</b>Compared with the healthy elderly subjects, the mild AD patients showed decreased ALFF in the right posterior cingulate cortex, right ventral medial prefrontal cortex, and in the bilateral dorsal medial prefrontal cortex. No brain region with increased ALFF was found in the AD group compared with the control group.</p><p><b>CONCLUSIONS</b>The reduced activity in the posterior cingulate cortex and medial prefrontal cortex observed in the present study suggest that the functional abnormalities of those areas are at an early stage of AD. The ALFF analysis may provide a useful tool in fMRI study of AD.</p>


Subject(s)
Aged , Alzheimer Disease , Diagnosis , Case-Control Studies , Female , Gyrus Cinguli , Humans , Magnetic Resonance Imaging , Methods , Male , Middle Aged , Prefrontal Cortex
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