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【Objective】 To investigate the protection of astragaloside IV from high glucose induced podocyte injury and mitochondrial dysfunction and its molecular mechanisms. 【Methods】 The model of podocyte injury induced by high glucose (30 mmol/L glucose) was established, and the model cells were treated with low, medium and high doses of astragaloside IV respectively; cell activity was detected by CCK-8. Apoptosis was detected by TUNEL staining. Mitochondrial membrane potential was detected by JC-1 fluorescence probe. ATP content was detected by the kit. The expression levels of apoptosis and podocyte injury related proteins and Notch pathway related proteins were detected by Western blotting. 【Results】 Compared with the control group, cell activity was decreased, apoptosis level was increased (P<0.05), anti-apoptotic protein (Bcl2) expression was decreased, and apoptosis protein (Bax, cleaved-caspase 9, cleaved-caspase 3) expressions were increased (all P<0.05) in HG group. Compared with HG group, HG+AS-IV improved cell activity and apoptosis level induced by high glucose (P<0.05), increased expression of anti-apoptotic protein (Bcl2), and decreased expressions of apoptotic protein (Bax, cleaved-caspase 9, and cleaved-caspase 3) (all P<0.05). Compared with the control group, mitochondrial dysfunction occurred in HG group, JC-1 monomer content increased, and ATP content decreased (all P<0.05). Compared with HG group, HG+AS-IV improved mitochondrial dysfunction, increased JC-1 polymer content and ATP content (P<0.05). In addition, compared with the control group, the expression of Notch pathway-related protein was decreased in HG group (P<0.05). Compared with HG group, Notch pathway-related protein expression was increased in HG+AS-IV group (all P<0.05). Molecular docking results showed that AS-IV could bind Notch1. 【Conclusion】 Astragaloside IV can improve podocyte injury and mitochondrial dysfunction induced by high glucose, possibly by inhibiting Notch pathway activation.
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Objective To observe the effect of anti NRP-1 b1/b2 monoclonal antibody (NRP-1mAb) on migration and invasion of gastric cancer cell line BGC-823, and to explore the possible mechanism. Methods NRP-1mAb was prepared in the laboratory, and the purity of antibody was detected by flow cytometry. The different concentrations of NRP-1mAb were added into the culture medium of gastric cancer cell line BGC-823. The migration and invasion of cells after 12 hours was observed by using Transwell method. The phosphorylation of related signal proteins after NRP-1mAb was detected by Western blot analysis. Results When NRP-1mAb prepared by patented technology had the effects on BGC-823 cells after 12 hours, the number of migration and invasion of BGC-823 cells was reduced. The number of cells through the basement membrane in the control group (blank) and the administration group (NRP-1mAb 25, 100, 400 μg / ml) were 167 ± 9, 138 ± 5, 98 ± 5, 36 ± 4, respectively (F = 22.6, P< 0.01); the number of cells through the filtration membrane were 231 ± 40,224 ± 19,176 ± 26,124 ± 34,respectively(F=26.63,P<0.01). There were statistically significant differences between the administered group and the control group at 100 and 400 μg/ml (all P< 0.001). High concentration of NRP-1mAb (100 μg/ml) decreased the phosphorylation level of Akt after 10 minutes' function on gastric cancer cells. However, it was difficult to detect phosphorylated Akt after 30 minutes. Conclusion NRP-1mAb may inhibit the migration and invasion of gastric cancer cell line BGC-823 by decreasing the phosphorylation of Akt, which is positively correlated with the concentration.
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<p><b>PURPOSE</b>To explore the impact of subtropical maritime monsoon climate on the frequency of ambulance use for trauma patients in a coastal region in China.</p><p><b>METHOD</b>Statistical analysis of data on ambulance use from the 120 Emergency Command Center in Shantou City, Guangdong Province, from January to December 2012 as well as daily meteorological data from a Shantou observatory was performed to determine how climatic factors (seasons, time, and weather) affect the frequency of ambulance use for trauma patients.</p><p><b>RESULTS</b>The daily ambulance use for trauma patients differed between spring and summer or autumn (p<0.05), between sunny and rainy days (p<0.05), and between cloudy and lightly or moderately rainy days (p<0.05).We found a linear correlation between daily maximum temperature and daily ambulance use for trauma patients (R² =0.103, p<0.05). In addition, there was significant difference in ambulance use between good and bad weather (p<0.05).</p><p><b>CONCLUSION</b>Frequency of ambulance use for trauma patients is affected by the subtropical maritime monsoon climate in the coastal region. Better weather contributes to increased daily frequency of ambulance use, which is the highest in autumn and lowest in spring.</p>
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Humans , Ambulances , China , Climate , Retrospective Studies , Wounds and Injuries , TherapeuticsABSTRACT
Objective This study is to explore the differences in the curative effect of intra -abdominal infusion between Recombinant mutant human tumor necrosis factor and Dendritic cell -Cytokine induced killer . Methods We selected 48 advanced gastric cancer patients with ascites .Those patients were randomized into two groups:DC-CIK group and rmhTNF group .After one month treatment ,we observed the adverse events both two groups,and evaluated the clinical beneficial responses ,the responsive rate ,the tumor indicators′level,the immune indexes and the time of tumor progression .Results In the DC-CIK group,the RR and the CBR were 83.3%and 66.67%,respectively;while in the rmhTNF group,they were 58.33%and 75%,respectively.The difference between two groups was statistically significant (P0.05).The ratios of CD3 +、CD4 +、NK cells distinctly increased after treatment (P0.05).One year follow-up of the time to progression(TTP)was 7.1 months in DC-CIK group,and 5.8 months in TNF group,which was statistically significant (P=0.02).Conclusion Both rmhTNF and DC-CIK can improve the efficacy for the patients with malignant ascites caused by gastric cancer .However ,DC-CIK immunotherapy shows better active effect on improvement of specific immune function .
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Objective To study the influence of survivin targetedly inhibited with antisense oligonucleotide (ASODN)technique on the apoptosis of hepatoma carcinoma cells SMMC-7221,and to clarify the mechanism of promotion effect of survivin-ASODN on the apoptosis of SMMC-7721 cells.Methods The ASODN sequence of survivin marked by FAM fluorescein was designed and synthized. The SMMC-7721 cells were transfected by different concentrations (100,200,300,400,and 600 nmol· L-1 )of survivin-ASODN (ASODN transfection groups),at the same time blank control group and blank liposome control group and sense oligonucleotide (SODN) control group were set up.The apoptotic rates and the changes of cell cycle of the SMMC-7721 cells 24,48,and 72 h after transfected with different concentrations of survivin-ASODN were detected by FCM. The expression levels of survivin were measured by Western blotting method.Results Compared with each control group,24 h after transfection,the apoptotic rates of survivin-ASODN transfected SMMC-7221 cells were increased,the growth of cells was inhibited (P<0.05),and the effects had time-dose dependent tendency.48 h after transfection,the hypodiploid apoptotic peak appeared in ASODN transfection groups before G1 phase, the number of the cells at G0/G1 phase was decreased (P<0.05)and the number of the cells at G2/M phase wsa increased (P<0.05). Compared with each control group,the survivin expression levels in the SMMC-7721 cells in ASODN transfection groups were decreased (P<0.05 ), and the effects of survivin-ASODN was time-dose dependent (P<0.05 ). Conclusion Survivin-ASODN can block the expression of survivin in SMMC-7721 cells and inhibit the proliferation of SMMC-7721 cells by changing the cell cycle and increasing apoptosis in a time-dose dependent manner.
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Objective: This study aimed to compare and analyze the functional differences between peripheral blood mono-cyte-derived dendritic cells (DCs) of Helicobacter pylori-positive and H. pylori-negative patients with gastric cancer. Methods:H. py-lori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the 14C-urea breath test. DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4, rhGM-CSF, and rhTNF-α. Furthermore, the expression of surface marker molecules was determined by fluorescence-activat-ed cell sorting analysis. The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydroge-nase-releasing assay. The secretion of IL-12 and IFN-γin the supernatant was determined by enzyme-linked immunosorbent assay. Re-sults:No difference was observed in the morphological change of the maturation process. The mean expression of CD1a, CD80, CD83, CD86, and HLA-DR molecules in DCs of H. pylori-infected patients was higher than that in DCs of H. pylori-negative group, and the differences were statistically significant except for CD1a and HLA-DR. The cytotoxicity activities, IL-12 release, and IFN-γrelease in the H. pylori-positive group were significantly higher than those in the H. pylori-negative group (P<0.05). Conclusion:H. pylori infec-tion has no effect on the morphological change of the maturation process of monocyte-derived DCs. These data clearly demonstrate that monocyte-derived DCs of H. pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H. pylori-negative patients.
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Dendritic cells (DCs) are the important auxiliary cells in tumor vaccines to activate antitumor immunity.Suppressor of cytokine signaling 1 ( SOCS1 ) has been shown to play an important role in inhibition of cytokine signal transduction.Recent studies show that inhibition of the function of SOCS1 in DCs plays a critical role in strongly enhancing tumor vaccines antitumor activity,and in the methods of restraining SOCS1 's function have also made significant progress.
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Objective To investigate whether gemcitabine (GEM) could enhance radiosensitivity of human non-small cell lung cancer cells and its related mechanism.Methods Clonogenic assay was used to analyze radiosensitivity enhancement by GEM on p53 mutant human lung adenocarcinoma cell line 973.Alterations of cell cycle distribution and apoptosis were measured by flow cytometry.Results Mild radiosesitizing effect was observed when 10 nmol/L GEM was administrated before or after irradiation.Marked radiosesitizing effect was demonstrated when 100 nmol/L GEM was administrated before or after irradiation, with much stronger effect of pre-irradiation GEM treatment.Mutation of p53 gene affected cell cycle redistribution and cell apoptosis, but had no relationship with radiosensitivity enhancement of GEM.Conclusions 100 nmol/L GEM could significantly enhance radiosensitivity of human lung cancer cells.However, this effect may not be associated with p53 gene mutation, cell cycle redistribution or cell apoptosis.
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Objective To establish the method of testing immunoglobulin G (IgG) with Fc sialylation, and to investigate the clinical significance of IgG with Fc sialylation in systemic lupus erythematosus (SLE), especially in those with neuropsychiatric manifestations (NPSLE). Methods Seventy-five SLE including thirty patients with neuropsychiatric manifestations (NPSLE) and forty-five non-NPSLE patients, 30 rheuma-toid arthritis (RA) patients, 32 juvenile idiopathic arthritis (JIA) patients and 41 healthy controls were recruited in this study. Standard method of testing lgG with Fc sialylation was established by a lectin based sandwiched enzyme linked immunosorbent assay (ELISA). The levels of IgG with Fc sailylation were assayed, and its clinical significance was evaluated. Results There were no difference in the levels of serum IgG with Fc sailylation in RA group (0.82±1.81) mg/ml, JIA group (0.69±1.30) mg/ml, healthy control group (0.64± 1.09) mg/ml, and the levels of IgG with Fc sailylation in SLE group (0.12±0.17) mg/ml (P<0.01), especially in NPSLE group [(0.03±0.03) mg/ml, P<0.01] was significantly lower than that of control groups. The perc-entage of IgG with Fc sialylation in control group (4.64±5.90)% were significantly higher than that in non-NPSLE (1.88±2.16)% (P<0.01) and in NPSLE (0.29±0.47)% (P<0.01). The percentage of IgG with Fc sial-ylation was negatively associated with SLEDAI score (r=-0.43, P<0.01). Conclusion Significantly low level of serum IgG with Fc sialylation was associated with disease activity in SLE patients, especially in NPSLE patients, lgG with Fc sialylation may be a new target for therapeutic strategy.
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<p><b>OBJECTIVE</b>To investigate the role of retinoic acid receptor beta (RARbeta) in mediating inhibitory effect of all-trans retinoic acid (ATRA) on activator protein-1 (AP-1) activity in gastric cancer cells.</p><p><b>METHODS</b>Transient transfection and chloramphenicol acetyltransferase (CAT) assay, Nort hern blot, gene transfection, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and anchorage independent growth assay were used.</p><p><b>RESULTS</b>Transient transfection of RARbeta expression vector into MKN-45 cells resulted in the RARbeta concentration dependent repression of AP-1 activity induced by 12-o-tetradecanoylphorbol-13-acetate (TPA), regardless of the presence of ATRA. When the c-jun and c-fos expression vectors were cotransfected with the RARbeta expression vector into MKN-45 cells, AP-1 activity was also obviously repressed. The inhibitory effect, again, was RARbeta-concentration-dependent. The stable transfection of the RARbeta gene into MKN-45 cells led to cell growth inhibition and colony formation inhibition by ATRA. Furthermore, Cotransfection of both RARbeta/DNA binding domain (DBD) and reporter gene could not alter AP-1 activity, even in the presence of ATRA.However, when the cotransfection was substituted with the RARbeta/ligand binding domain (LBD), the inhibition was significantly enhanced by ATRA.</p><p><b>CONCLUSION</b>RARbeta might be required for anti-AP-1 activity, and contribute to growth inhibition of gastric cancer cells by ATRA.</p>
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Binding Sites , Cell Division , DNA , Metabolism , Receptors, Retinoic Acid , Chemistry , Physiology , Stomach Neoplasms , Drug Therapy , Pathology , Transcription Factor AP-1 , Tretinoin , Pharmacology , Tumor Cells, CulturedABSTRACT
PURPOSE To investigate the effects of all trans retinoic acid(ATRA) on the growth of gastric cancer cells and explore the mechanism relevant to enzyme.METHODS With MTT method the growth inhibition of gastric cancer cells was measured;the activity of lactate dehydrogenase(LDH),alkaline phosphatase(ALP) and ? glucuronidase(? G) was measured with metaplate;The activity of succinate dehydrogenase(SDH) was assayed with cytochemical method.RESULTS The growth of MGc80 3, but not MKN 45, was inhibited,which is dependent on the concentration of ATRA.The activity of LDH,ALP and ? G of Mgc80 3 was suppressed and the activity of SDH of Mgc80 3 was enhanced.But all the four enzymes′ activity of MKN 45 were not obviously changed except that introcelluar ? G activity was inhibited.CONCLUSIONS The sensitivity of gastric cancer cells to ATRA is different,which is closely related with the change of enzyme activity.
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The endothelial cells activated by tumor cells are genetically stable and regress easily. Moreover, at the surface there are specific markers, which are easily accessible by system administration. This report review the studies of tumor treatment targeting at the activated endothelial cells.