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1.
Article in Chinese | WPRIM | ID: wpr-883824

ABSTRACT

Objective:To screen the time points of high survival rate and efferocytosis dysfunction of rat alveolar macrophages stimulated by cigarette smoke extract (CSE), establish an in vitro model of alveolar macrophage efferocytosis function, and study chronic respiratory diseases with chronic inflammatory reaction as the main pathological changes. Methods:① Time point screening experiment: rat alveolar macrophages (NR8383 cells) were cultured in vitro, and the cells in logarithmic growth phase were divided into blank control group (100 μL complete medium) and 5% CSE group (90 μL complete medium + 10 μL 100% CSE). Alma blue method was used to detect the effect of 5% CSE on the activity of NR8383 cells at 6, 12, 24 and 48 hours. ② Apoptosis induction experiment: rat type Ⅱ alveolar epithelial cells (RLE-6TN cells) were cultured in vitro as phagocytic target cells of NR8383 cells, and the cells in logarithmic growth phase were divided into blank control group and 10, 30 and 60 minutes groups after ultraviolet exposure (apoptosis was induced by 30 000 μJ/cm 2 ultraviolet irradiation for 15 minutes). Flow cytometry was used to detect the apoptosis rate of RLE-6TN cells cultured for 10, 30 and 60 minutes after ultraviolet exposure. ③ Cell efferocytosis experiment: NR8383 cells in logarithmic phase were divided into blank control group and 5% CSE group. Two hours before NR8383 cells were stimulated by CSE for 6, 12 and 24 hours, RLE-6TN cells were exposed to ultraviolet to induce apoptosis, and the RLE-6TN cell suspension was added to NR8383 cells (the ratio of RLE-6TN cells to NR8383 cells was 5∶1). Flow cytometry was used to detect the efferocytosis rate of NR8383 cells to RLE-6TN cells at different time points treated with 5% CSE. Results:① Compared with the blank control group, the activity of NR8383 cells significantly decreased after treatment with 5% CSE for 48 hours [cell reduction rate: (68.5±4.1)% vs. (73.6±2.3)%, P < 0.05]. However, there were no significant differences when the activities of NR8383 cells treated with 5% CSE for 6, 12 and 24 hours were compared with the blank control group, so these three time points were selected for the subsequent establishment of alveolar macrophage cell efferocytosis dysfunction in vitro model experiment. ② Compared with the blank control group, the apoptosis rate of RLE-6TN cells significantly increased at 10, 30 and 60 minutes after ultraviolet exposure [(66.87±8.63)%, (85.51±2.39)%, (96.13±2.74)% vs. (9.13±3.17)%, all P < 0.01] in a time-dependent manner. Considering that it taked about 50 minutes for RLE-6TN cells to be labeled with PKH26 membrane labeling probe, 10 minutes after ultraviolet exposure was selected to label RLE-6TN cells. ③ Compared with the blank control group, the efferocytosis function of NR8383 cells was significantly decreased after treatment with 5% CSE for 12 hours [cell efferocytosis rate: (33.64±1.30)% vs. (44.02±2.71)%, P < 0.01], but there was no significant effect on the efferocytosis function of NR8383 cells at 6 hours and 24 hours. Conclusions:CSE can induce alveolar macrophage cell efferocytosis dysfunction. Based on the test results of the effect of 5% CSE on NR8383 cell activity and cell efferocytosis function, 12 hours with high survival rate and weak efferocytosis effect of NR8383 cells can be selected as the in vitro model condition of alveolar macrophage cell efferocytosis dysfunction.

2.
Article in Chinese | WPRIM | ID: wpr-828936

ABSTRACT

OBJECTIVE@#To investigate the protective effect of melatonin against myocardial ischemia reperfusion (IR) injury in isolated rat hearts and explore the underlying mechanisms.@*METHODS@#The isolated hearts from 40 male SD rats were randomly divided into 4 groups (=10): the control group, where the hearts were perfused with KH solution for 175 min; IR group, where the hearts were subjected to global ischemia for 45 min followed by reperfusion for 120 min; IR+melatonin (Mel+IR) group, where melatonin (5 μmol/L) was administered to the hearts 1 min before ischemia and during the first 5 min of reperfusion, followed by 115 min of reperfusion; and IR+2, 3-butanedione monoxime (IR+BDM) group, where the hearts were treated with BDM (20 mmol/L) in the same manner as melatonin treatment. Myocardial injury in the isolated hearts was assessed based on myocardial injury area, caspase-3 activity, and expressions of cytochrome C and cleaved caspase-3 proteins. Cardiac contracture was assessed using HE staining and by detecting lactate dehydrogenase (LDH) activity and the content of cardiac troponin I (cTnI) in the coronary outflow, measurement of left ventricular end-diastolic pressure (LVEDP) and electron microscopy. The content of ATP in the cardiac tissue was also determined.@*RESULTS@#Compared with those in the control group, the isolated hearts in IR group showed significantly larger myocardial injury area and higher caspase-3 activity and the protein expressions of cytochrome C and cleaved caspase-3 with significantly increased LDH activity and cTnI content in the coronary outflow and elevated LVEDP at the end of reperfusion; HE staining showed obvious fractures of the myocardial fibers and the content of ATP was significantly decreased in the cardiac tissue; electron microscopy revealed the development of contraction bands. In the isolated hearts with IR, treatment with Mel or BDM significantly reduced the myocardial injury area, caspase-3 activity, and protein expressions of cytochrome C and cleaved caspase-3, obviously inhibited LDH activity, lowered the content of cTnI and LVEDP, reduced myocardial fiber fracture, and increased ATP content in the cardiac tissue. Both Mel and BDM inhibited the formation of contraction bands in the isolated hearts with IR injury.@*CONCLUSIONS@#Mel can alleviate myocardial IR injury in isolated rat hearts by inhibiting cardiac contracture, the mechanism of which may involve the upregulation of ATP in the cardiac myocytes to lessen the tear of membrane and reduce cell content leakage.


Subject(s)
Animals , Contracture , Male , Melatonin , Myocardial Ischemia , Myocardial Reperfusion Injury , Myocardium , Myocytes, Cardiac , Rats , Rats, Sprague-Dawley
3.
Article in Chinese | WPRIM | ID: wpr-828517

ABSTRACT

OBJECTIVE@#To investigate the protective effect of melatonin against myocardial ischemia reperfusion (IR) injury in isolated rat hearts and explore the underlying mechanisms.@*METHODS@#The isolated hearts from 40 male SD rats were randomly divided into 4 groups (=10): the control group, where the hearts were perfused with KH solution for 175 min; IR group, where the hearts were subjected to global ischemia for 45 min followed by reperfusion for 120 min; IR+melatonin (Mel+IR) group, where melatonin (5 μmol/L) was administered to the hearts 1 min before ischemia and during the first 5 min of reperfusion, followed by 115 min of reperfusion; and IR+2, 3-butanedione monoxime (IR+BDM) group, where the hearts were treated with BDM (20 mmol/L) in the same manner as melatonin treatment. Myocardial injury in the isolated hearts was assessed based on myocardial injury area, caspase-3 activity, and expressions of cytochrome C and cleaved caspase-3 proteins. Cardiac contracture was assessed using HE staining and by detecting lactate dehydrogenase (LDH) activity and the content of cardiac troponin I (cTnI) in the coronary outflow, measurement of left ventricular end-diastolic pressure (LVEDP) and electron microscopy. The content of ATP in the cardiac tissue was also determined.@*RESULTS@#Compared with those in the control group, the isolated hearts in IR group showed significantly larger myocardial injury area and higher caspase-3 activity and the protein expressions of cytochrome C and cleaved caspase-3 with significantly increased LDH activity and cTnI content in the coronary outflow and elevated LVEDP at the end of reperfusion; HE staining showed obvious fractures of the myocardial fibers and the content of ATP was significantly decreased in the cardiac tissue; electron microscopy revealed the development of contraction bands. In the isolated hearts with IR, treatment with Mel or BDM significantly reduced the myocardial injury area, caspase-3 activity, and protein expressions of cytochrome C and cleaved caspase-3, obviously inhibited LDH activity, lowered the content of cTnI and LVEDP, reduced myocardial fiber fracture, and increased ATP content in the cardiac tissue. Both Mel and BDM inhibited the formation of contraction bands in the isolated hearts with IR injury.@*CONCLUSIONS@#Mel can alleviate myocardial IR injury in isolated rat hearts by inhibiting cardiac contracture, the mechanism of which may involve the upregulation of ATP in the cardiac myocytes to lessen the tear of membrane and reduce cell content leakage.


Subject(s)
Animals , Heart , Male , Melatonin , Pharmacology , Therapeutic Uses , Muscle Contraction , Myocardial Reperfusion Injury , Drug Therapy , Myocytes, Cardiac , Rats , Rats, Sprague-Dawley
4.
Chinese Critical Care Medicine ; (12): 913-915, 2019.
Article in Chinese | WPRIM | ID: wpr-754079

ABSTRACT

As one of the top three causes of death in the world, chronic obstructive pulmonary disease (COPD) is a serious hazard to human health. Macrophages play an important role in COPD, and their efferocytosis function is essential for ending chronic inflammation of COPD. Efferocytosis damage of alveolar macrophages (AM) in patients with COPD causes the rising of bacterial infection and airway bacterial colonization risk in lungs, which is the main reason for the acute exacerbation and the rising of incidence rate and mortality rate in COPD. In recent years, the regulation of macrophage efferocytosis function in COPD has becoming a research hotspot. Progress on the role of macrophage efferocytosis function on COPD, and the breakthrough points of improving AM efferocytosis dysfunction by traditional Chinese medicine is reviewed, so as to provide new ideas for the prevention and treatment of COPD.

5.
Article in Chinese | WPRIM | ID: wpr-693772

ABSTRACT

Objective:To investigate the role of Na+/Ca2+ exchanger (NCX) in myocardial ischemiareperfusion injury and the underlying mechanisms.Methods:Forty Sprague-Dawley rats were divided into 4 groups randomly:a control group,a KBR7943 group,an ischemia-reperfusion group (IR group),and an IR plus KB-R7943 group (KB-R7943+IR group).Isolated Sprague Dawley male rat hearts underwent Langendorffperfusion.The ratio of left ventricular developed pressure (LVDP),left ventricular end-diastolic pressure (LVEDP),the infarct size of myocardium,and the lactate dehydrogenase (LDH) activity in the coronary flow was determined.HE staining was used to assess the change of myocardial morphology.Western blot was used to determine the levels of cleaved caspase-3,cytochrome c and the phosphorylation of Ca2+/calmodulin-dependent protein kinase Ⅱ (CaMKⅡ) and the Thr17 site ofphospholamban.Results:Compared with the control group,IR group significantly induced an enlarged infarct size,reduction of the ratio of LVDP,up-regulation of cytochrome c,cleaved caspase-3,p-CaMKⅡ and p-phospholamban,and increased in the activity of LDH,the level of LVEDP (P<0.01) and the disordered myocardial morphology.These effects were significantly attenuated in the presence of KB-R7943 treatment (10 μmol/L).Conclusion:NCX mediates myocardial ischemia-reperfusion-induced cell apoptosis and necrosis through activation of CaMKⅡ.

6.
Article in Chinese | WPRIM | ID: wpr-614461

ABSTRACT

Objective To investigate the influence of three different processing methods on contents and speciation of heavy metals Pb,Cd,Hg and Cu and a harmful nonmetallic element As in Cortex Eucommiae. Methods The five elements were measured by atomic absorption spectrometry. The determination wavelengthes of Pb and Cd was 283.3 and 228.8 nm using graphite furnace atom?ic absorption spectrometry;the determination wavelength of Cu was 324.8 nm using flame atomic absorption spectrometry;the determi?nation wavelengthes of As and Hg were 193.7 and 253.6 nm using flow injection-hydride generation method. Results Heavy metal′s contents in Cortex Eucommiae stir-fried with salt were increased. Contents of heavy metals in Cortex Eucommiae stir-fried and baked were decreased. Heavy metal morphology had no differences between processed products and raw materials. The experimental results showed that main existing form of Pb was in organic matter-bound,and mercury existed in exchange condition and organic matter-bound. Copper existed in organic matter-bound and carbonate bound and arsenic mainly existed with residual forms. Cadmium mainly exited in carbonate exchange condition. Conclusion There is a certain effect on the heavy metal content but less effect on the form of heavy metals by different processing methods.

7.
Article in Chinese | WPRIM | ID: wpr-513559

ABSTRACT

Objective To observe the effects of nourishing lung and kidney formulas on inflammatory response of alveolar epithelial cells stimulated by monocytes conditioned medium and study the anti-inflammatory mechanism of the formulas for the treatment of chronic obstructive pulmonary diseases (COPD).Methods The reproduction of inflammation models of A549 cells were stimulated by monocyte THP-1 cell strain conditioned medium.A549 cells were randomly divided into blank control group (20% blank rabbits serum),model group (20% blank rabbits serum+25.0% THP-1 cell conditioned medium),nuclear transcription factor activator protein-1 (AP-1) pathway inhibitor T-5224group (20% blank rabbits serum+25.0% THP-1 cells conditioned medium+100 μmol/L T-5224),nourishing lung and kidney group (20% rabbits serum with nourishing lung and kidney formulas+25.0% THP-1 cells conditioned medium).The contents of interleukins (IL-6,IL-8),tumor necrosis factor-α (TNF-αα),matrix metalloprotein-9 (MMP-9) in cell culture supernatant were detected with enzyme linked immunosorbent assay (ELISA),the supernatant content of malondialdehyde (MDA) was detected with thibabituric acid (TBA) method,the total activity of superoxide dismutase (T-SOD) was detected with hydroxylamine method,and the activity of AP-1 pathway was detected with electrophoretic mobility shift assay (EMSA) method.Results Compared with the blank control group,the A549 cell proliferation were significantly increased at 24 hours,48 hours stimulation by 25.0% cell conditioned medium (A value:24 hours was 0.41 ± 0.02 vs.0.37 ± 0.04,48 hours was 1.30 ± 0.09 vs.1.15 ± 0.19).Compared with the blank control group,the contents of IL-6,IL-8,TNF-αα,MMP-9,MDA,AP-1 expression were significantly increased in model group [IL-6 (ng/L):35.00±3.63 vs.23.15±1.72,IL-8 (ng/L):273.09± 164.36 vs.231.45±33.90,TNF-α(ng/L):51.61 ± 9.51 vs.28.87 ± 3.34,MMP-9 (ng/L):442.85 ± 78.86 vs.235.60 ± 14.62,MDA (μmol/L):6.90 ± 0.11 vs.6.01 ± 0.12,AP-1 expression (A value):2.260 ± 0.062 vs.1.000 ± 0.000],MDA/T-SOD ratio was increased (4.43 ± 0.05vs.3.96 ± 0.06).Compared with model group,the levels of IL-8 (ng/L:100.29 ± 17.03),TNF-α (ng/L:25.13 ± 0.46),AP-1 expression (A value:1.38 ± 0.02),and the MDA/T-SOD ratio (4.23 ± 0.23) in T-5224 group,and MMP-9 (ng/L:195.44±9.80),MDA (μmol/L:5.86±0.30),MDA/T-SOD ratio (3.56±0.41),AP-1 expression (A value:0.76 ± 0.01) in nourishing lung and kidney group were all reduced significantly (all P < 0.05).Conclusion Nourishing lung and kidney formulas can suppress the inflammatory response through regulating the alveolar epithelial cells AP-1 signaling pathways.

8.
Article in Chinese | WPRIM | ID: wpr-491779

ABSTRACT

Objective:To study the related factors of severe acute radiation-induced lung injury (SAR) caused by IMRT and concurrent chemotherapy for non-small cell lung cancer. Methods:We retrospectively analyzed the data of 2 323 non-small cell lung cancer pa-tients who underwent IMRT radiotherapy and concurrent chemotherapy at the Department of Radiotherapy of Tianjin Medical Univer-sity Cancer Institute and Hospital from January 2010 to January 2014. We analyzed the clinical factors and parameters that affect dose by univariate and multivariate analysis. Results:A total of 2 323 patients enrolled and 1 241 cases suffering from acute radiation-in-duced lung injury with the rate of 53.4%. Only 185 cases suffered from SARP with a rate of 7.96%. Univariate analysis showed that the gender, histopathological type, total radiation dose, V5 (%), and average dose rate are not related to SARP (P>0.05). By contrast an age of>60 years, 1%predicted FEV, docetaxel+carboplatin/cisplatin chemotherapy, V20 (%), V30 (%), and mean lung dose (MLD) are sig-nificantly related to SARP (P60 years, docetaxel+carboplatin/cisplatin che-motherapy, V20 (%), and V30 (%) are the independent risk factors of SARP. Conclusion:Among the non-small cell lung cancer patients undergoing IMRT radiotherapy and concurrent chemotherapy, further attention should be given to elderly patients, patients receiving docetaxel and platinum chemotherapy, as well as V20 and V30 with high doses. The necessary preventive treatment should be given to reduce the incidence of SARP, improve the quality of life of patients, and reduce the incidence of respiratory failure and mortality.

9.
Chinese Critical Care Medicine ; (12): 765-768, 2016.
Article in Chinese | WPRIM | ID: wpr-497308

ABSTRACT

Cell culture technology is the most commonly used method in the in vitro experiments at present. However, monolayer cell culture technology has been unable to meet the demand of the researchers. This is because that monolayer cell culture cannot mimic the cellular environment in which multiple cells interact with each other in the body. We cannot discuss the relationship of many cells, because we do not know the relationship between cells through a single kind of cell. So cell co-culture medicine arises at the historic moment for the demand. With the development of research method in recent years, cell co-culture method also has been improved in practice: from direct contact co-cultures to indirect contact co-cultures, from two-dimensional co-cultures to three-dimensional co-cultures. Cell co-culture method is closer to the human body. It is also more advantageous to study the interaction among cells. Nowadays, there are more researchers tend to select this method to study the physiological and pathological in vitro model, tissue engineering, and cell differentiation research. At the same time, it has become the focus of drug research and development, drug analysis, mechanism of drug action, and drug targets. This article will review the studies of cell co-culture method, summarize advantages and disadvantages of various methods, so as to promote improvement of cell culture methods, to build cells co-culture system that more close to human body, and build the in vitro model that simulate internal circulation of human body further.

10.
Article in Chinese | WPRIM | ID: wpr-476267

ABSTRACT

Objective The aim of this study was to generate human cholesteryl ester transfer protein ( CETP) transgenic rabbits and analyze their biological properties.Methods We generated human CETP transgenic rabbits by DNA microinjection, and detected the expression of human CETP by real-time PCR and Western blot assay.The activity of CETP was measured using an activity assay kit.Results Human CETP transgenic rabbits were successfully generated by DNA microinjection.Compared with wide type rabbits, the expression of human CETP was dramatically increased in the liver of the human CETP transgenic rabbits.The plasma CETP activity was also much higher in the liver of human CETP transgenic rabbits than that of control rabbits.Conclusions The model of human CETP transgenic rabbits is successfully established by DNA microinjection.It will provide a useful tool for the studies of CETP biological function and its involvement in the mechanisms of cardiovascular diseases.

11.
Article in Chinese | WPRIM | ID: wpr-469409

ABSTRACT

Objective To observe the clinical efficacy of duloxetine in the treatment of malignant neuropathic pain with depression.Methods 60 patients were randomly divided into two groups as study group(30 cases) and contrdl group(30 cases) and treated for 4 weeks.The patients of study group were treated with duloxetine and oxycontin,and the patients of control group were treated with oxycontin only.Numberical rating scale (NRS) on pain,criteria of pain relief and Hamilton depression scale(HAMD,17 items) score were used to assess the therapeutic effect before and after treatment.Results By the end of the fourth week of treatment,the average usage of oxycontin of the study group was significantly less than control group((45.6±8.5) mg vs (88.2±5.2)mg,P<0.05).The effective rate of pain relief in the study group was significantly higher than that in control group (93.3% vs 73.3%,P<0.05).Comparing pre-treatment,the score of HAMD of the study group had a remarkable decrease ((11.45±4.56) vs (23.07±5.47),P<0.01).In comparison to the score of control group,study group had a significant effect ((11.45±4.56) vs (18.75±4.21),P<0.01).Conclusion Duloxetine is one of effective agents in the treatment of malignant neuropathic pain with depression,which can alleviate depression and relieve pain.Duloxetine have mild adverse effects and good tolerance.

12.
Article in Chinese | WPRIM | ID: wpr-464697

ABSTRACT

Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.

13.
Article in Chinese | WPRIM | ID: wpr-461191

ABSTRACT

ABSTRACT:Objective To analyze the correlation between plasma nitric oxide (NO)level and atherosclerotic lesion in high cholesterol-fed (HCD ) rabbits.Methods Twenty male Japanese white rabbits were divided randomly into two groups,which were fed with normal diet (control group,n =10)or HCD (experimental group, n =10 )for 1 6 weeks.At the end of the experiment,plasma lipid and NO levels were measured.The gross atherosclerotic lesions in each group were detected by Sudan IV staining while intimal lesion area was measured by hematoxylin/eosin (HE)and elastica van gieson (EVG).Moreover,the macrophages (MΦ)and smooth muscle cells (SMC)were detected by immunohistochemical staining.The correlation analysis was made to reveal the relationship between atherosclerotic lesions and plasma NO level.Results Compared with those in control group, the total cholesterol (TC),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C)and NO levels all increased significantly in experimental group.Atherosclerotic lesions appeared on the vascular wall in the latter group.The area of atherosclerotic lesions and MΦ in the plaque had a positive association with plasma NO level. Conclusion There is a relationship between plasma NO level and the size of HCD-induced atherosclerotic lesions in rabbits.Meanwhile,the MΦ positive area in the atherosclerotic plaque is also associated with plasma NO level in cholesterol-fed rabbits,suggesting that plasma NO level may be associated with the occurrence and progress of early atherosclerosis.

14.
Article in Chinese | WPRIM | ID: wpr-460737

ABSTRACT

Objective:To explore the feasibility and safety of CT-guided hookwire localization of small lung nodule in video-as-sisted thoracic surgery. Methods: Preoperative localization of small lung nodule was performed using the CT-guided hookwire tech-nique, followed by video-assisted thoracic surgery in the wedge resection. The next mode of operation depends on the results of frozen biopsy. Results:Preoperative localization with CT-guided hookwire was performed in 34 patients between February 2012 and March 2014. The diameter of lung nodule ranged from 5 mm to 22 mm. CT-guided hookwire localization was successful in all patients, with a median positioning time of 23 min. Puncture needles were detached from two of the total patients during the surgery, and three other pa-tients showed pneumothorax by CT scan after localization. Conclusion:Preoperative hookwire localization of small lung nodule is an accurate and safe approach to improve the rate of wedge resection in video-assisted thoracic surgery.

15.
Article in Chinese | WPRIM | ID: wpr-487182

ABSTRACT

Objective To study the effect of Qige San ethyl acetate extract on the proliferation of human esophageal carcinoma Eca109 cells and to explore its possible mechanism. Methods Inhibitory effects of Qige San ethyl acetate extract on the proliferation of esophageal carcinoma Eca109 cells were detected by MTT assay, cell apoptosis was detected by flow cytometer, and the expression of protein STAT3, Bcl-2 and Caspase 9 was detected by Western blotting method. Results In the range of 10~100 μg/mL, Qige San ethyl acetate extract inhibited the proliferation of Eca109 cells effectively (P<0.05) . Compared with the control group, Qige San ethyl acetate extract in the concentrations of 1.47, 33.26, 75.52 μg/mL significantly increased the apoptotic rate of Eca109 cells within 48h ( P<0.01). And Western blotting results showed that the ex pression levels of STAT3 and Bcl-2 were reduced, and Caspase 9 was increased with the increase of drug concentration. Conclusion Qige San ethyl acetate extract could significantly inhibit the proliferation of Eca109 cells and induce cell apoptosis, and its mechanism is probably associated with the inhibition of signal transducers and activators of transcription 3 (STAT3) signaling pathway.

16.
Chinese Journal of Oncology ; (12): 651-656, 2014.
Article in Chinese | WPRIM | ID: wpr-272316

ABSTRACT

<p><b>OBJECTIVE</b>The aim of this study was to investigate the effects of combination of icotinib and cetuximab on the acquired drug resistance caused by T790M mutation of EGFR in NSCLC, and provide experimental evidence for rational treatment of NSCLC.</p><p><b>METHODS</b>The effects of these two agents on cell proliferation, apoptosis, and EGFR-dependent signaling were evaluated using 3-(4, 5-dimethylthiazol-2-yl)- 5-diphenyltetrazolium bromide (MTT) assay, annexin V staining, and Western blotting. The expression of molecular markers of tumor proliferation PCNA and Ki-67 protein was further examined by immunohistochemistry, and the expression of EGFR-signaling-related proteins in tissue sections taken from H1975 tumor xenografts was assessed by Western blot assay. Sensitivity to EGFR inhibitors was detected in human H1975 tumor xenograft in nude mice.</p><p><b>RESULTS</b>The in vitro experiment showed that the proliferative ability of H1975 cells was inhibited in a dose-dependent manner, along with the increasing doses of cetuximab and icotinib, and the combination of cetuximab with icotinib resulted in a more pronounced growth inhibition of the H1975 cells. The apoptosis rate of H1975 cells after treatment with 0.5 µmol/L icotinib and 1 µg/ml cetuximab was (22.03 ± 2.41)% and that after treatment with 5 µmol/L icotinib and 10 µg/ml cetuximab was (42.75 ± 2.49)%, both were significantly higher than that after treatment with the same dose of icotinib or cetuximab alone (P < 0.05). The nude mouse experiment showed that the transplanted tumor was growing to (614.5 ± 10.8) mm(3) in the blank control group and to (611.2 ± 8.7) mm(3) at 28 days after icotinib treatment, but (30.8 ± 2.0) mm(3) in the cetuximab treatment group and 0 mm(3) in the cetuximab combined with icotinib group. There was a significantly decreased expression of Ki-67 and PCNA proteins and down-regulation of phosphorylation of EGFR signaling-related proteins in the cetuximab combined with icotinib group.</p><p><b>CONCLUSIONS</b>The combination of icotinib with cetuximab can exert synergistic inhibitory effect on the acquired drug resistance caused by T790M mutation of EGFR in NSCLC H1975 cells, interrupts the EGFR-downstream signaling pathway, and enhances the anticancer activity of chemotherapeutic drugs. Our results provide further experimental evidence for the clinical studies of combination of icotinib with cetuximab in the treatment of NSCLC patients associated with secondary drug resistance caused by T790M mutation of EGFR.</p>


Subject(s)
Animals , Antibodies, Monoclonal, Humanized , Therapeutic Uses , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Apoptosis , Carcinoma, Non-Small-Cell Lung , Drug Therapy , Genetics , Cell Line, Tumor , Cell Proliferation , Cetuximab , Crown Ethers , Therapeutic Uses , Down-Regulation , Drug Resistance, Neoplasm , Genetics , Genes, erbB-1 , Genetics , Humans , Lung Neoplasms , Drug Therapy , Mice , Mice, Nude , Mutation , Quinazolines , Therapeutic Uses , ErbB Receptors , Signal Transduction
17.
Chinese Journal of Digestion ; (12): 446-450, 2013.
Article in Chinese | WPRIM | ID: wpr-437078

ABSTRACT

Objective To explore the efficacy of flupenthixol melitracen in the treatment of functional dyspepsia (FD) and its effects on receptivity expansion and visceral sensitivity.Methods A randomized,controlled,multi-centre study was carried out.The FD patients with depression were divided into anti-depression group and control group.The patients of both groups received omeprazole and mosapride treatment for eight weeks.The patients of anti-depression group added flupenthixol melitracen and psychotherapy.Before the treatment,by the end of the second,fourth,sixth and eighth week of the treatment,the scores of upper gastrointestinal symptoms and adverse effects of the patients of the two groups were recorded,the psychological status was evaluated by Hamilton depression scale and the gastric receptivity expansion and visceral sensitivity were assessed by water loading test (WLT).The data were analyzed by t test and chi-square test.Results Among two hundred and fifty enrolled FD patients with depression,245 patients (129 in anti-depression group,116 in control group) were enrolled in intention-to treat analysis.Before the treatment and by the end of the eighth week of the treatment,the scores of upper gastrointestinal symptoms of anti depression group were 12.23±4.45 and 2.31±2.13 (t=22.837,P=0.008),the control group were 12.29± 4.62 and 8.58±3.76 (t=6.708,P=0.184).By the end of the eighth week of treatment,the difference between the two groups was statistically significant (t =16.261,P<0.05).By the end of the fourth and eighth week of treatment,the total effective rates of the improvement of upper gastrointestinal symptoms of anti-depression group were 76.0% (98/129) and 83.7% (108/129),control group were 14.7 % (17/116) and 25.9 % (30/116),the differences between the two groups were statistically significant (x2 =92.191 and 83.117,both P<0.05).By the end of the eighth week of treatment,the total effective rate of the improvement of depression of anti depression group was 71.3% (92/129),which was higher than that of control group (23.3%,27/116),the difference between two groups was statistically significant (x2 54.434,P<0.01).Before the treatment,the maximum WLT volume of anti depression group was (850±130) mL,by the end of the eighth week of treatment was (1400±210) mL (t=25.292,P<0.01).Before the treatment and by the end of the eighth week of treatment,the maximum WLT volumes of control group were (800± 120) mL and (960±140) mL (t=9.345,P<0.01).All the adverse effects of anti depression therapy were mild.Conclusion The treatment of flupenthixol melitracen can improve the efficacy in patients with FD,relieve the depression status,improve gastric receptivity expansion and visceral sensitivity with a good safety and tolerability.

18.
Article in Chinese | WPRIM | ID: wpr-397817

ABSTRACT

Objective To investigate the effect of the psychological behavior intervention on the mental status and clinical symptom of patients with functional constipation. Methods All of 78 patients with functional constipation diagnosed by Rome m diagnostic code were randomly divided into study group (n =40) and control group (n = 38). The psychological behavior intervention was used in the study group, the caccagogue and symptomatic treatment was used in the control group. The self-rating anxiety scale(SAS),self-rating depressive scale (SDS) was analyzed and the improving of clinical symptom, adverse effect was compared before and after 4 weeks of treatment respectively. Results The scores of SAS and SDS in the study group significantly decreased after treatment [ (36.25±8.61 )scores vs (42.97±9.25 )scores, P<0.01 ;(38.81±6.53)scores vs (43.36±7.18 )scores ,P < 0.01].There was significant difference between the two groups after treatment (P <0.05).The effective rate of study group was higher than that of control group .(97.50% vs 84.21%,P <0.05). The rate of celiodynia, diarrhea and the recrudescence after the course of treatment in control group was 23.68%, 15.79% and 89.47% respectively, while this numerus in study group was 5.00%,2.50% and 5.00%. Conclusions Psychological behavior intervention plays an important role in the treatment of functional constipation. It can improve the mental status, clinical symptom, and decrease the adverse effect.

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Article in Chinese | WPRIM | ID: wpr-584230

ABSTRACT

Objective:To evaluate the efficacy of paroxetine on functional low fever. Methods:Using a randomized and placebo controlled trail, a total of 54 with functional low fever were randomized into paroxetine group and placebo group. In paroxetine group, 28 patients received oral paroxetine 1 tablet (20mg/tablet), one times a day for 8 weeks. There were 26 patients in the placebo. The criteria of assessing the therapeutic efficacy on functional low fever and Hamilton Depression Scale (HAMD; 17 items) were used to assess the therapeutic efficacy at the 4th and 8th weekend respectively.Results:After 8 weeks of treatment, the efficacy rate on functional low fever and antidepression were 78.6%, 82.1% in paroxetine group and 26.9%, 23.1% in the placebo group respectively. There was a significant difference between the two groups (P

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Article in Chinese | WPRIM | ID: wpr-578861

ABSTRACT

Objective To investigate the effects and possible mechanisms of ursolic acid (UA) on inhibiting proliferation of human esophageal carcinoma cell Eca-109 and inducing its apoptosis. Methods Cell proliferation was determined by MTT assay. Electron microscope was used to observe the ultrastructural changes of Eca-l09 induced by UA. Cell cycle and apoptotic rate were analyzed by flow cytometry (FCM),and the expression of P27kip1,Bcl-2 and Bax were detected by Western blot method. Results UA could significantly inhibit the growth of Eca-109 cells(P

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