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OBJECTIVE To explore the neuroprotective effect and possible mechanism of celastrol (Cel) and its derivatives (Cel-1, Cel-2) in terms of neuroinflammation and oxidative damage. METHODS Neuroinflammation model of microglial BV2 cells was induced by 1 μg/mL lipopolysaccharide (LPS); oxidative damage model of human neuroblastoma SH-SY5Y cells was induced by 200 μmol/L hydrogen peroxide (H2O2). The toxicity of different concentrations of Cel, Cel-1 and Cel-2 (0.625-20 μmol/L) to the two types of cells was investigated. The levels of nitric oxide (NO), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in BV2 cells induced by LPS at safe concentrations (0.039-0.625 μmol/L) were all detected. The survival rate of SH-SY5Y cells induced by H2O2 was also determined. The expression levels of phosphoinositide 3-kinase (PI3K), p-PI3K, protein kinase B (Akt), p-Akt, cystatinase 3 (caspase-3), B-cell lymphoma 2 (Bcl-2) and Bcl-2-related X protein (Bax) in SH- SY5Y cells induced by H2O2 at 0.156, 0.313, 0.625 μmol/L of active compound 2 were all detected. RESULTS In the concentration gradient range between 0.039 and 0.625 μmol/L, the results of neuroinflammation model experiments showed that Cel, Cel-1 and Cel-2 could reduce the contents of NO, TNF-α, IL-1β, and IL-6 in culture medium of BV2 cells (P<0.05 or P< 0.01); their IC50 values for neuroinflammation were (0.25±0.04), (0.61±0.14) and (0.11±0.02) μmol/L respectively. Meanwhile, all of them could reverse the phenomenon of decreased cell survival rate after H2O2 treatment in the oxidative damage experiments at a certain concentration (P< 0.05 or P<0.01), with neuroprotective EC50 values of (0.43± XJC2023009) 0.08), (0.45±0.04) and (0.28±0.03) μmol/L, respectively.Induced by H2O2, the phosphorylation of PI3K and Akt protein, protein expressions of Bcl-2 and Bcl-2/Bax ratio were all increased significantly (P<0.05 or P<0.01), while the protein expressions of caspase-3 and Bax were decreased significantly (P<0.05 or P<0.01). CONCLUSIONS Cel, Cel-1, and Cel-2 all have significant neuroprotective activities at certain concentrations, and Cel-2 shows the most significant protective effect. The mechanism of action of Cel-2 may be related to regulating the PI3K/Akt and caspase-3/Bcl-2/Bax signaling pathways, reducing the inflammatory response, oxidative stress damage and inhibiting neuronal apoptosis.
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ObjectiveTo investigate the effects of Linggui Zhugantang on mitochondrial fission and fusion and silencing information regulator 3(Sirt3)/adenosine monophosphate dependent protein kinase (AMPK) signaling pathway in chronic heart failure (CHF) rats after myocardial infarction (MI). MethodSD rats randomly divide into sham operation group (normal saline ,thread only without ligature), model group (normal saline, ligation of the left anterior descending coronary artery proximal to the heart), Linggui Zhugantang group (4.8 g·kg-1) and Captopril group (0.002 57 g·kg-1), with 10 rats in each group. Administere drug continuously for 28 days. Echocardiography detected cardiac function parameters. Hematoxylin eosin (HE) staining observed the pathological changes of the heart. Immunofluorescence detected the levels of reactive oxygen species (ROS). JC-1 detect mitochondrial membrane potential. Colorimetry measure adenosine triphosphate (ATP), superoxide dismutase (SOD), malondialdehyde (MDA), mitochondrial respiratory chain complex activity (Ⅰ-Ⅳ). TdT-mediated dUTP nick end labeling (TUNEL) staining detected the apoptosis rate of myocardial tissue. Western blot detected protein expression levels of Sirt3, phosphorylated AMPK (p-AMPK), phosphorylated dynamic-related protein 1(p-Drp1), mitochondrial fission protein 1(Fis1), mitochondrial fission factor (MFF), optic atrophy protein 1(OPA1). ResultCompared to the sham group, the left ventricular end diastolic diameter (LVIDd) and left ventricular end systolic diameter (LVIDs) were significantly increased in model group (P<0.01), while the left ventricular short axis shortening rate (LVFS) and left ventricular ejection fraction (LVEF) were significantly decreased (P<0.01). There were inflammatory cell infiltration and obvious pathological injury in myocardial tissue. ROS, MDA levels and myocardial cell apoptosis rate were significantly increased (P<0.01), SOD level, ATP content, and membrane potential were significantly decreased (P<0.01). The activity of mitochondrial respiratory chain complexes (Ⅰ-Ⅳ) was significantly decreased (P<0.01). Levels of p-Drp1, Fis1, MFF proteins were significantly up-regulated (P<0.01), while Sirt3, p-AMPK, OPA1 proteins level were significantly down-regulated (P<0.01). Compared with model group, LVIDd and LVIDs were significantly decreased (P<0.01), LVEF and LVFS were significantly increased (P<0.01). Inflammatory cell infiltration and pathological damage of myocardial tissue were significantly relieved. ROS, MDA levels and myocardial cell apoptosis rate were significantly decreased in Linggui Zhugantang group and Captopril group (P<0.01), SOD level, ATP content, and membrane potential significantly increased (P<0.01). The activity of mitochondrial respiratory chain complexes (Ⅰ-Ⅳ) increased significantly (P<0.01),and p-Drp1, Fis1, MFF protein levels were significantly down-regulated (P<0.01), Sirt3, p-AMPK, OPA1 protein were significantly up-regulated (P<0.01). ConclusionLinggui Zhugantang can alleviate oxidative stress and apoptosis damage of myocardial cells, maintain mitochondrial function stability, and its effect may be related to mitochondrial mitosis fusion and Sirt3/AMPK signaling pathway.
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Angelicae Sinensis Radix (AS) is reproted to exert anti-depression effect (ADE) and nourishing blood effect (NBE) in a rat model of depression. The correlation between the two therapeutic effects and its underlying mechanisms deserves further study. The current study is designed to explore the underlying mechanisms of correlation between the ADE and NBE of AS based on hepatic metabonomics, network pharmacology and molecular docking. According to metabolomics analysis, 30 metabolites involved in 11 metabolic pathways were identified as the potential metabolites for depression. Furthermore, principal component analysis and correlation analysis showed that glutathione, sphinganine, and ornithine were related to pharmacodynamics indicators including behavioral indicators and hematological indicators, indicating that metabolic pathways such as sphingolipid metabolism were involved in the ADE and NBE of AS. Then, a target-pathway network of depression and blood deficiency syndrome was constructed by network pharmacology analysis, where a total of 107 pathways were collected. Moreover, 37 active components obtained from Ultra Performance Liquid Chromatography-Triple-Time of Flight Mass Spectrometer (UPLC-Triple-TOF/MS) in AS extract that passed the filtering criteria were used for network pharmacology, where 46 targets were associated with the ADE and NBE of AS. Pathway enrichment analysis further indicated the involvement of sphingolipid metabolism in the ADE and NBE of AS. Molecular docking analysis indciated that E-ligustilide in AS extract exhibited strong binding activity with target proteins (PIK3CA and PIK3CD) in sphingolipid metabolism. Further analysis by Western blot verified that AS regulated the expression of PIK3CA and PIK3CD on sphingolipid metabolism. Our results demonstrated that sphingolipid metabolic pathway was the core mechanism of the correlation between the ADE and NBE of AS.
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Rats , Animals , Rats, Sprague-Dawley , Molecular Docking Simulation , Network Pharmacology , Drugs, Chinese Herbal/chemistry , Metabolomics/methods , Mass SpectrometryABSTRACT
Objective:To investigate the efficacy and safety of bevacizumab combined with capecitabine in the treatment of advanced breast cancer.Methods:Seventy-six patients with advanced breast cancer who were diagnosed in the Cancer Center of the People's Liberation Army Navy Anqing Hospital from August 2019 to May 2021 were selected. According to different treatment schemes, the patients were divided into the control group (using single drug capecitabine) and the test group (using bevacizumab combined with capecitabine), with 38 cases in each group. After 4 cycles of treatment, the clinical efficacy, progression-free survival (PFS), overall survival (OS) and adverse reactions were compared between the two groups, and the levels of vascular endothelial growth factor (VEGF) -121, VEGF-145, VEGF-165 and quality of life before and after treatment were compared.Results:The objective remission rate of the test group [57.89% (22/38) ] was higher than that of the control group [42.11% (16/38) ], but there was no statistically significant difference ( χ2=1.89, P=0.169) ; The disease control rate of the test group [81.58% (31/38) ] was better than that of the control group [55.26% (21/38) ], there was a statistically significant difference ( χ2=6.09, P=0.014). The median PFS of patients in the test group (6.3 months) was longer than that in the control group (4.2 months), there was a statistically significant difference ( χ2=0.48, P=0.003) ; The median OS of patients in the test group (14.8 months) was not significantly different from that in the control group (13.2 months) ( χ2=0.15, P=0.704). After treatment, the expression level of serum VEGF-121 [ (201.25±18.37) ng/L vs. (276.83±20.26) ng/L], VEGF-145 [ (102.24±12.16) ng/L vs. (170.39±15.28) ng/L], VEGF-165 [ (135.08±14.32) ng/L vs. (210.53±16.09) ng/L] in the test group was lower than that in the control group, there were statistically significant differences ( t=17.03, P<0.001; t=21.51, P<0.001; t=21.59, P<0.001). After treatment, patients in the test group were assessed according to 36-item Short-Form (SF-36) physiological function [ (80.18±13.96) score vs. (71.72±16.12) score], physiological function [ (67.19±30.62) score vs. (53.12±9.86) score], physical pain [ (70.01±17.97) score vs. (61.06±17.57) score], overall health [ (68.67±18.92) score vs. (57.96±20.97) score], vitality [ (78.39±19.37) score vs. (68.26±18.52) score], social function [ (82.24±19.73) score vs. (70.92±20.31) score], the scores of emotional function [ (73.81±28.86) score vs. (60.23±29.19) score] and mental health [ (76.19±12.82) score vs. (70.31±12.54) score] were higher than those of the control group, there were statistically significant differences ( t=2.45, P=0.017; t=2.03, P=0.046; t=2.19, P=0.031; t=2.34, P=0.022; t=2.33, P=0.023; t=2.46, P=0.016; t=2.04, P=0.045; t=2.02, P=0.047). The incidence of adverse reactions in the test group [18.42% (7/38) ] was lower than that in the control group [76.32% (29/38) ], there was a statistically significant difference ( χ2=25.54, P<0.001) . Conclusion:The combination of bevacizumab and capecitabine chemotherapy has a higher clinical effect on advanced breast cancer, which can significantly reduce the level of VEGF in patients, improve the quality of life of patients, with mild adverse reactions and high safety.
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BACKGROUND: Lymph node ratio (LNR) has been suggested to be an effective prognostic tool for stratifying non-small cell lung cancer (NSCLC) cases. In this study, we sought to determine cancer-specific survival (CCS) of NSCLC cases from the SEER registry and used the X-tile method to optimize CCS-based LNR cut-off points for prognostic stratification of node-positive NSCLC. METHODS: CSS and other clinicopathologic variables were retrieved from the SEER registry. Kaplan-Meier methods were used to calculate CSS. The optimal cut-off points for LNR classification were determined by the X-tile approach. Multivariate Cox regression analysis was performed to identify independent risks of CSS. RESULTS: Totally 11,341 lung cancer patients were included. Their median CSS was 22 months (range 0,143). The median LNR was 0.22 (Q1,Q3: 0.11, 0.50). X-tile analysis showed that the optimal LNR cut-off points were 0.28 and 0.81, dividing the cohort into low (LNR1 ≤ 0.28; n = 6580, 58%), middle (0.28 < LNR2 < 0.81; n = 3025, 26.7%), and high (LNR3 > 0.81; n = 1736, 15.3%) subsets. Kaplan-Meier analysis showed that patients with a low LNR had a significantly higher CCS versus patients with middle or high LNR (P < 0.001). Multivariate competing risks regression analysis revealed that LNR was an independent and significant adverse predictor of CSS (LNR2 vs. LNR1: SHR: 1.56, 95%CI: 1.47,1.67, P < 0.001; LNR3 vs. LNR1: SHR: 2.54, 95%CI: 2.30,2.80, P < 0.001). CONCLUSIONS: LNR is an independent prognostic factor of node-positive NSCLC and its optimal cut-off values established using the robust x-tile method effectively define subpopulations of node-positive NSCLC cases, which is important in guiding selection of treatment strategies clinically.
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Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/mortality , Lung Neoplasms/pathology , Lymph Node Ratio , Adolescent , Adult , Aged , Aged, 80 and over , Carcinoma, Non-Small-Cell Lung/diagnosis , Female , Humans , Kaplan-Meier Estimate , Lung Neoplasms/diagnosis , Lymphatic Metastasis , Male , Middle Aged , Multivariate Analysis , Neoplasm Staging , Prognosis , Proportional Hazards Models , Retrospective Studies , SEER Program , Young AdultABSTRACT
Objective To investigate the association between the nutritional status and related factors in patients with Parkinson's disease (PD).Methods Seventy-two patients with PD (PD group) and 71 age-and sex-matched healthy controls (control group) were enrolled in this study from September 2014 to November 2017 at the First Affiliated Hospital of Sun Yat-sen University.Their serum nutritional indices,including serum albumin,prealbumin,transferrin,free fatty acid and retinol conjugated protein,were collected.The PD participants were interviewed and assessed using motor and non-motor scales,including Hoehn and Yahr stage,Movement Disorder Society-Unified Parkinson's Disease Rating Scale (MDS-UPDRS),Non-Motor Symptoms Questionnaire for Parkinson's Disease (NMSS),Mini-Mental State Examination (MMSE),Unified Dyskinesia Rating Scale (UDysRS) and 39-item Parkinson's Disease Questionnaire (PDQ-39).Their medication dosage was indicated by daily levodopa equivalent doses (LEDD).Body mass index (BMI) was used to determine their nutritional status,with abnormal nutritional status defined as BMI lower than 20 kg/m2.Results Levels of serum albumin (41.60 (40.28,43.98) g/L vs 44.00 (42.30,46.20) g/L,Z=4.500,P<0.01),transferrin ((2.32±0.34) g/L vs (2.51±0.34) g/L,t=-3.305,P=0.001),and free fatty acid (418.00 (289.75,637.25) μmol/L vs 547.00 (386.00,699.00) μmol/L,Z=2.079,P=0.038) of the PD group were significantly lower than those of the control group.There was a significant negative correlation between serum albumin and MDS-UPDRS-Ⅱ score (r=-0.254,P=0.031),MDS-UPDRS-Ⅳ score (r=-0.256,P=0.030),years of dyskinesia (r=-0.240,P=0.043),years of motor fluctuation (r=-0.304,P=0.009) and LEDDs (r=-0.321,P=0.006).Disease duration was negatively correlated with serum albumin (r=-0.285,P=0.015) and transferrin (r=-0.275,P=0.019),and age (r=-0.252,P=0.032) was negatively correlated with prealbumin.The forward binary Logistic regression model indicated that abnormal nutritional status was closely associated with rigidity (OR=1.171,95%CI 1.013-1.354,P=0.032),akinesia (OR=1.070,95%CI 1.000-1.144,P=0.048),UDysRS score (OR=1.051,95%CI 1.004-1.099,P=0.032),MDS-UPDRS-Ⅳ score (OR=1.177,95%CI 1.018-1.360,P=0.027) and MMSE score (OR=0.821,95%CI 0.678-0.994,P=0.043),but not correlated with tremor and axial symptoms.Compared with PD patients with abnormal nutritional status,PD patients with normal nutritional status had higher MMSE scores (28.00 (27.00,29.00) vs 28.00 (25.00,28.00),Z=-2.060,P=0.039),lower rigidity (9.60±3.83 vs 12.00±4.29,t=-2.264,P=0.027),akinesia (19.98 ± 8.00 vs 24.42:±:8.06,t=-2.071,P=0.042) and MDS-UPDRS-Ⅳ scores (8.00 (5.00,11.00) vs 10.00 (9.00,13.00),Z=2.642,P=0.008).Conclusions PD patients tend to have a lower serum nutritional indices.PD patients with lower levels of serum nutritional indices are characterized by more severe motor complication,longer disease duration,older age and higher LEDD.PD patients with abnormal nutritional status have worse cognition and more severe motor symptoms (rigidity,akinesia and motor complication).
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Objective To investigate CT and MRI features of hepatic sclerosed hemangioma (HSH).Methods CT and MRI findings were retrospectively reviewed in 20 cases of HSH,all of which were confirmed pathologically after hepatic surgery.Twenty patients underwent CT scan,4 patients underwent MRI.Meanwhile,the enhancement pattern and signal intensity were analyzed either.Results Twenty patients showed main part of tumor was hypo-attenuating on CT plain scanning,and 16 patients showed the central area of tumor was markedly more hypo-attenuating on CT plain scanning.After administration of intravenous contrast media,multifocal linear or small nodular enhancement in the peripheral area was seen during the arterial phase on 16 patients of HSH.Venous phases showed centripetal enhancement or spread around the nodules enhancement which continued to delayed phases with low density of no enhancement in the lesion area.Four patients showed no obvious enhancement on arterial phases and slight separation sample enhancement at the edge or inside of the lesions with a wide range of non enhancement areas on venous phase and delayed phase.Four patients were performed MRI examination,the lesions demonstrated hypointensity with a lower signal area on T1WI,hyperintensity with a higher signal intensity area on T2WI.The DWI sequence of b value were 0,150,800 s/mm2,all of which were obviously hypointensity.The edge of lesions showed small nodular enhancement on arterial phase,irregular concentric enhancement on venous phase and delayed phase,and there was no enhancement area with lower signal in the center of the lesion.Conclusions The enhancement pattern of HSH different from cavernous hemangioma,with a larger non enhancement area in the center of the lesions and similar to other hepatic masses with central scar,differential diagnosis dependence on CT and MRI dynamic enhanced scan.
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<p><b>OBJECTIVE</b>To construct a lentiviral vector carrying the γ-synuclein(SNCG) gene and establish a human colorectal carcinoma cell line SW1116 stably expressing this gene, and then investigate the inhibition of the growth and invasion capacity of SW1116 cells.</p><p><b>METHODS</b>RNA interference fragment was designed according to the SNCG sequence (GenBank: No.NM003087.2), and then SNCG RNAi effective target genes were screened. After the Oligo DNA of target sequences was synthesized, the lentiviral vectors carrying LV-SNCG-RNAi-EGFP (RNAi group) and LV-SNCG-NC-EGFP (NC group) were constructed and packaged to produce lentivirus venom. The supernatants of different virus-producing cells were used to transfect SW1116 cells respectively. Wild SW1116 cells were used as blank control (CON group) EGFP fluorescence was detected by fluorescent microscopy and the differential expression of SNCG mRNA and protein was detected by real-time PCR and Western blot. CCK-8, soft agar assay and Transwell chamber were employed to estimate the inhibiting effect on growth and invasion of SW1116 respectively.</p><p><b>RESULTS</b>Recombinant lentiviral vectors respectively carrying the SNCG-RNAi-EGFP and SNCG-NC-EGFP were successfully constructed and the supernatants of lentivirus could effectively infect SW1116 cells. The titer of the virus carrying LV-SNCG-RNAi-EGFP or LV-SNCG-NC-EGFP was 8×10(8) TU/ml. Real-time PCR and Western blot confirmed that compared with the NC group, SNCG-RNAi group had lower SNCG expression (1.009±0.161 vs. 0.114±0.030, P=0.009), and showed tremendous silencing effect as 76.8%(P<0.05). SNCG protein expression was also significantly reduced (RNAi:12.001±2.884, NC:32.443±4.731, CON:34.308±6.920, P<0.05). After SNCG knockdown, the number of proliferation cells was obviously reduced at 48, 72, 96 and 120 hours respectively(P=0.036). In soft agar assay, clones in RNAi group were smaller[RNAi:(0.582±0.103) mm, NC:(1.863±0.316) mm, CON:(1.749±0.525) mm]. Colony formation rate of RNAi group was down to (17.1±3.5)%, which was significantly lower than (36.5±4.3)% in NC group and (33.8±3.9)% in CON group. In migration test, the number of invasion cell was 37.4±9.3 in RNAi group, which was significantly less than 112.3±8.6 in NC group and 100±0.0 in CON group.</p><p><b>CONCLUSION</b>Expression of SNCG mRNA and protein plays an important role in the growth and the invasion capacity of SW1116 cells.</p>
Subject(s)
Humans , Cell Line, Tumor , Cell Proliferation , Colorectal Neoplasms , Pathology , Genetic Vectors , Lentivirus , RNA Interference , RNA, Messenger , RNA, Small Interfering , Genetics , Real-Time Polymerase Chain Reaction , Transfection , gamma-Synuclein , GeneticsABSTRACT
Objective To investigate the clinical effect of longxuejie capsule in combination with tinidazole in the treatment of chronic cervicitis and its effects on levels of immunoglobulin.Methods 90 cases in our hospital from January 2013 to June 2014, with chronic cervicitis meeting the inclusion critera were randomly divided into 2 groups by method of stratified randomization equally.Control group (n=45) was treated with tinidazole and conventional treatments, while observation group ( n =45 ) was treated with tinidazole and longxuejie capsule.Then, the curative efficacy, scale of symptoms and levels of immunoglobulin were observed and compared.Results The therapeutic efficiency ratio of observation group was 95.6%, which was significantly higher than that of control group ( 80.0%, P<0.05 ).After the treatment, in comparison with the control group, scale of cervical secretion and severity of cervical lesion was statistically lower respectively than observation group ( P<0.05 ).As to immunological function, concentrations of IgG, IgM, IgA in observation group were statistically higher than that in control group after the treatment (P<0.05).The control group and observation group respectively in 2 cases and 3 cases of abdominal pain, alleviated by themselves without treatment.Conclusion longxuejie capsule combined with tinidazole is effective and reliable for patients with chronic cervicitis, which significantly improves symptoms and elevates levels of immunoglobulin.
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Objective To explore a method for establishing the hamster model of Clostridium difficile-associated diarrhea (CDAD)and the indicators for its evaluation.Methods Clindamycin was administered to hamsters subcutaneously (day 1),and 24 h later infected with C.difficile clinical isolates KH1 (ribotype 027,106-108 CFU/mL)or SH9 (ribotype 001 ,108-1010 CFU/mL)by gavage.Animals were observed for CDAD symptoms such as diarrhea,weight loss and death.At the end of ob-servation period (day 7 or death),the cecum was collected from each animal for histological evaluation of inflammation.Results Following a single dose of 100 mg/kg clindamycin subcutaneously,all the animals challenged with KH1 (108 CFU/mL)devel-oped diarrhea and then died within 5 days.All the hamsters challenged with SH9 (1010 CFU/mL)developed diarrhea as well but only 66.7% died at the end of observation period.Among other groups,only one or none developed diarrhea and then died. The symptoms of hamsters with diarrhea included loose stool,wet tail and weight loss.On histological examination,conges-tion,hemorrhage and neutrophil infiltration of the mucosa were observed in the hamsters died of CDAD.Conclusions We have successfully established a hamster CDAD model that allows for future investigations.
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Lignocellulosic biomass can be utilized to produce promising biofuels. In this study, liquid hot water pretreatments were performed to break the intricate structure of sugarcane bagasse, which resists the enzyme accessibility to cellulose. The effects of temperatures and times on the hemicellulose degradation (including the yields of pentoses and hexoses, the proportion of monomers and oligosaccharides, as well as limited inhibitors) and cellulose enzymatic digestibility were evaluated. The results indicated that the maximum xylose yields (combined 3.85 g xylose and 13.21 g xylo-oligosaccharides per 100 g raw material) in prehydrolyzate liquid were obtained at 180 °C and 30 min. Due to the effective removal of hemicellulose, the maximum glucose yield in enzyme hydrolyzate reached 37.27 g per 100 g raw material, representing 90.13% of glucose in the sugarcane bagasse. The maximal total sugars yield (combined prehydrolyzate and enzymatic hydrolyzate) were 53.65 g based on 100 g raw material.
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Carbohydrates/chemistry , Enzymes/chemistry , Hot Temperature , Saccharum/chemistry , Water , HydrolysisABSTRACT
ObjectiveTo provide reference for the prevention and treatment of common chronic diseases,and to discuss the relationship between disease and lifestyle.Methods Residents survey Questionnaire was made.All the residents from Capital Airport Community has been investigated.Results The top ten diseases in this area were:hypertension,diabetes,coronary heart disease,dyslipidemia,osteoporosis,cervical spondylosis,chronic bone and joint disease,hemorrhoids,cataracts,and benign prostatic hyperplasia.ConclusionChronic disease and lifestyle of the inhabitants from the region are closely related.
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To investigate the transgenic expressing efficacy of helper-dependent adenoviral vector (HDAd) in vitro, we constructed a HDAd encoding enhanced green fluorescent protein (EGFP), denominated as HDAd/EGFP, performed large scale preparation and purification, and then identified the purified HDAd/EGFP under fluorescent microscope and electron microscope. After the concentration of HDAd/EGFP was determined by spectrophotometer, the transgenic expression efficiency of HDAd/EGFP was compared with first generation adenoviral vector encoding EGFP (FGAd/EGFP) in vitro. Therefore, we infected A549 cells with 2000 virus particles (vp) per cell by HDAd/EGFP and FGAd/EGFP respectively and analyzed EGFP expressing level by flow cytometry. Consequently, the fluorescent expression rate and fluorescent intensity of EGFP were higher in early infected A549 cells by HDAd/EGFP than by FGAd/EGFP. HDAd, capable of expressing transgene instantly and efficiently in vitro, is a potential vaccine vector.
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Humans , Adenoviridae , Genetics , Metabolism , Cell Line, Tumor , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Helper Viruses , Genetics , Metabolism , Transgenes , Viral Fusion Proteins , Genetics , MetabolismABSTRACT
Purpose To investigate the role of urokinase plasminogen activator (uPA),uPA receptor (uPAR),tissue type plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) in the invasiveness of human breast cancer cells. Methods Three human breast cancer cell lines with different invasive ability were taken as research targets.RT-PCR and milk plates methods were used to detect the expression of uPA system members and the PA activities,respectively.Modified Boyden's chamber model was employed to detect the invasive ability of cancer cell. Results MDA-MB-231 could express high level of uPA,uPAR,PAI-1 and low level of tPA.MDA-MB-435 could express lower level of uPA and hight level of tPA,but no PAI-1 and uPAR were detected.MCF-7 could express lower level of uPAR and high level of PAI-1,but no uPA and tPA were detected.MDA-MB-231 cells showed the highest total PA and uPA activity.MDA-MB-435 cells also showed high total PA activity,but almost all the activity owed to tPA.MCF-7 showed almost no PA activity.Correlated with their PA activities,MDA-MB-231 was found the most invasive in vitro,followed by MDA-MB-435,and MCF-7 almost had no invasive ability.The antibodies against uPA and uPAR were significantly effective in reducing the matrigel invasiveness of MDA-MB-231 by approximately 83.1% and 43.9% respectively (P<0.05). Conclusions Co-expression of uPA,uPAR and PAI-1 in human breast cancer highly correlates with the invasiveness in vitro.