ABSTRACT
Background@#We found microRNA (miR)-1246 to be significantly differentially expressed between severe active alopecia areata (AA) patients and healthy individuals. @*Objective@#To explore the role and mechanism of miR-1246 in severe AA. @*Methods@#Expression of miR-1246, dual-specific tyrosine phosphorylation-regulated kinase 1A (DYRK1A), and nuclear factor of activated T cells 1c (NFATc1) in peripheral CD4+ T cells and in scalp tissues of patients were detected using RT-qPCR, Western blot, and immunohistochemistry assays. Peripheral CD4+ T cells from the AA patients were transfected with lentiviral vectors overexpressing miR-1246. RT-qPCR and Western blot analysis were used to measure mRNA or protein expression of retinoic-acid-receptor-related orphan nuclear receptor gamma (ROR-γt), interleukin (IL)-17, DYRK1A, NFATc1, and phosphorylated NFATc1. Flow cytometry was used to assay the CD4+ IL-17+ cells proportion. ELISA was used to measure cytokine levels. @*Results@#miR-1246 levels decreased and DYRK1A and NFATc1 mRNA levels significantly increased in the peripheral CD4+ T cells and scalp tissues of severe active AA samples.NFATc1 protein expression was also significantly increased in the peripheral CD4+ T cells but not in the scalp tissues. NFATc1 positive cells were mainly distributed among infiltrating inflammatory cells around hair follicles. In peripheral CD4+ T cells of severe active AA, overexpression of miR-1246 resulted in significant downregulation of DYRK1A, NFATc1, ROR-γt, and IL-17 mRNA and phosphorylated NFATc1 protein, as well as a decrease in the CD4+ IL-17+ cells proportion and the IL-17F level. @*Conclusion@#miR-1246 can inhibit NFAT signaling and Th17 cell activation, which may be beneficial in the severe AA treatment.
ABSTRACT
Objective: To systematically study the anti-fibrotic effect of N-acetyl-seryl-as partyl-lysyl-proline (Ac-SDKP) on pulmonary fibrosis. Methods: In May 2021, a computer search was performed on CNKI, Wanfang Knowledge Service Platform, VIP.com, China Biomedical Literature Database, Pubmed, OVID and other databases. The retrieval time was from January 2008 to May 2021. Randomized controlled experiments on the inhibition of pulmonary fibrosis by Ac-SDKP were screened. The control group was the pulmonary fibrosis model group and the experimental group was the Ac-SDKP treatment group. The quality of the literature was assessed using the syrcle risk of bias assessment tool, and data were extracted. Data analysis was Performed using revman 5.4 software. Results: 18 papers were included, with a total of 428 animal models. The results of meta analysis showed that the contents of α-smooth muscle actin (α-SMA), type I collagen, type Ⅲ collagen, transforming growth factor-β (TGF-β) and Nodule area in the exPerimental group were lower than those in the control grouP. [SMD=-2.44, 95%CI (-3.71--1.17), P=0.000][SMD=-5.36, 95%CI (-7.13--3.59), P=0.000] [SMD=-3.07, 95%CI (-4.13--2.02), P<0.000][SMD=-2.88, 95%CI (-3.63--2.14), P=0.000] [SMD=-1.80, 95%CI (-2.42--1.18), P=0.000], the content of hydroxy proline in the experimental group was higher than that in the control group [SMD=7.62, 95%CI (4.90-10.33), P=0.000], all indexes included in the literature were statistically significant. Conclusion: Ac-SDKP has obvious inhibitory effect on the process of pulmonary fibrosis, and may become a new clinical drug for the treatment of pulmonary fibrosis.
Subject(s)
Rats , Animals , Pulmonary Fibrosis , Rats, Wistar , Fibrosis , Disease Models, Animal , ProlineABSTRACT
Objective:To observe the influence of shared medical appointments on health-related quality of life and quality of sleep in patients after liver transplantation.Methods:By randomized controlled study, a total of 124 patients after liver transplantation were included from our hospital from January 2018 to January 2019, and according to the lottery method, all subjects were divided into the routine management group ( n=64) who received routine outpatient intervention and the shared medical management group ( n=60) who received shared medical appointments management. The health-related quality of life and quality of sleep were investigated and compared by post-liver transplant quality of life questionnaire (pLTQ) and Pittsburgh sleep quality index (PSQI) before intervention (the day of discharge) and after intervention (the end of the last shared outpatient service). Results:After intervention, the dimension scores of worry, economics, body function, emotional function, health service, complication and total score of pLTQ were improved in tow groups than before intervention [the routine management group: (41.90±7.61), (18.13±4.22), (22.22±5.10), (14.92±3.28), (20.39±4.87), (14.63±3.99), and (132.19±37.09) vs (32.25±5.55), (12.77±3.47), (17.58±4.72), (9.23±1.38), (15.17±4.81), (10.89±1.51) and (98.00±29.03) score, t=8.20, 7.85, 3.58, 12.79, 6.10, 7.01, 5.81, all P<0.001; shared medical management group: (46.12±7.92), (24.16±5.34), (25.55±5.42), (17.90±3.60), (24.81±5.12), (17.93±3.60) and (155.47±41.00) vs (32.57±5.69), (12.81±3.82), (17.00±4.70), (9.60±1.39), (15.39±4.84), (11.00±3.52) and (98.37±28.96) score, t=10.76, 13.39, 9.23, 16.66, 10.36, 10.66, 8.81, all P<0.001], and those in the shared medical management group were higher than those in routine management group ( t=3.03, 6.95, 3.53, 4.82, 4.93, 4.83, 3.32, all P<0.05). After intervention, the total score of PSQI scale were lower than before intervention in the routine management group [(10.48±2.14) vs (11.89±2.45) score, t=3.47, P=0.001], and the dimensions score of sleep quality, full-sleep time, sleep time, sleep efficiency, sleep disorders, daytime function, hypnotic and total score of PSQI were lower than before intervention in the shared medical management group [(1.41±0.32), (0.54±0.13), (1.17±0.26), (1.11±0.35), (1.21±0.27), (1.30±0.33), (1.08±0.21) and (8.05±1.75) vs (1.88±0.53), (0.86±0.37), (1.84±0.41), (2.05±0.56), (1.39±0.33), (1.47±0.43), (1.22±0.32) and (11.71±2.43) score, t=-5.88, -6.32, -10.69, -11.03, -3.27, -2.43, -3.65, -9.47, all P<0.05], and those in the shared medical management group were lower than those in routine management group ( t=-6.68, -6.39, -10.43, -10.97, -2.62, -2.12, -3.54, -6.90, all P<0.05). Conclusion:Shared medical appointments model can improve the health-related quality of life and quality of sleep in patients after liver transplantation, and improve the effectiveness of outpatient intervention.
ABSTRACT
Objective: To study the effect of anti-fibrotic tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) on phosphorylated heat shock protein 27 (P-HSP27) and zinc finger family transcriptional repressor 1 (SNAI1) expression to explore the anti-silicosis fibrosis effect of Ac-SDKP. Methods: In December 2014, the rat silicosis animal model was prepared by one-time bronchial infusion of silicon dioxide (SiO(2)) dust. 80 SPF healthy adult Wistar rats were selected, and the rats were divided into 8 groups according to the random number table method, 10 in each group. Model control group for 4 weeks (feeding for 4 weeks) , model control group for 8 weeks (feeding for 8 weeks) : bronchial perfusion with normal saline 1.0 ml per animal. Silicosis model group for 4 weeks (feeding for 4 weeks) and silicosis model group for 8 weeks (feeding for 8 weeks) : bronchial perfusion of 50 mg/ml SiO(2) suspension 1.0 ml per animal. Ac-SDKP administration group for 4 weeks (feeding for 4 weeks) , Ac-SDKP administration group for 8 weeks (feeding for 8 weeks) : Ac-SDKP 800 μg·kg(-1)·d(-1) was administered by intraperitoneal pump. Ac-SDKP preventive treatment group: 48 h after Ac-SDKP 800 μg·kg(-1)·d(-1) administration, bronchial perfusion of SiO(2) suspension 1.0 ml per animal, raised for 8 weeks. Ac-SDKP anti-fibrosis treatment group: after bronchial perfusion of 1.0 ml of SiO(2) suspension for 4 weeks, Ac-SDKP 800 μg·kg(-1)·d(-1) was administered for 4 weeks. Western blotting was used to detect the expression of P-HSP27, SNAI1, α-smooth muscle actin (α-SMA) , and collage typeⅠ and Ⅲ in each group. The expression of P-HSP27 and SNAI1 was detected by immunohistochemistry, and the co-localized expression of P-HSP27 and α-SMA was detected by laser confocal microscopy. Results: Compared with the model control group, the expressions of P-HSP27, SNAI1, α-SMA, and collage typeⅠ and Ⅲ in the silicosis fibrosis area of the rats in the silicosis model group were enhanced, and the differences were statistically significant (P<0.05) . After Ac-SDKP intervention, compared with silicosis model group for 8 weeks, the expressions of P-HSP27, SNAI1 α-SMA, and collage typeⅠ and Ⅲ in the Ac-SDKP preventive and anti-fibrosis treatment groups were significantly decreased, and the differences were statistically significant (P<0.05) . However, the expressions of P-HSP27 SNAI1, and collage typeⅠ and Ⅲ between the Ac-SDKP administration group and the model control group did not change significantly, and the differences were not statistically significant (P>0.05) . Laser confocal results showed that the positive cells expressing P-HSP27 and α-SMA in the lung tissue of the silicosis model group were more than those in the model control group. Compared with the silicosis model group, the Ac-SDKP prevention and anti-fibrosis treatment groups expressing the positive cells of P-HSP27 and α-SMA decreased. Compared with the model control group for 8 weeks, there were some double-positive cells expressing P-HSP27 and α-SMA in the nodules of the silicosis model group for 8 weeks. Conclusion: Ac-SDKP may play an anti-silicic fibrosis effect by regulating the P-HSP27/SNAI1 pathway.
Subject(s)
Animals , Rats , HSP27 Heat-Shock Proteins , Oligopeptides , Rats, Wistar , Silicon Dioxide , Silicosis/metabolismABSTRACT
Objective:To prepare bone marrow mesenchymal stem cells (BMSCs) transfected with therapeutic gene early growth reactive protein 1 (Egr1)-sodium/iodine symporter (NIS) and carrying gold nanoparticles (AuNPs), and to investigate the feasibility of Egr1 in promoting NIS expression and the radiasensitization effect of AuNPs.Methods:BMSCs transfected with lentivirus(Lv)-Egr1-NIS-cytomegalovirus(CMV)-green fluorescent protein (GFP) in the experimental group and Lv-Egr1-GFP in the control group were prepared and the expression of NIS induced by radioiodine was verified by iodine uptake determination. The optimal incubation time and concentration of AuNPs were observed with laser confocal microscopy. The cytotoxicity of AuNPs suspension liquid was investigated using cytotoxicity test. Iodine uptake assay was performed to investigate NIS gene expression of BMSCs-Egr1-NIS incubated with AuNPs. In vitro chemotaxis of BMSCs-Egr1-NIS incubated with/without AuNPs to breast cancer cells were verified by cell migration experiment. The radiosensitization effect of AuNPs for 131I on killing breast cancer cells MDA-MB-231 were explored. The one-way analysis of variance and Dunnett t test were used for data analysis. Results:BMSCs-Egr1-NIS (unstable transformation) was successfully prepared. Egr1 could promote NIS expression with the induction of radioiodine. The iodine uptake capacity in BMSCs-Egr1-NIS increased by 2.5-5 times or even higher compared with BMSCs-Egr1-GFP. The better incubation conditions of AuNPs for BMSCs phagocytosis were 0.20 g/L(24 h) or 0.10 g/L(48 h). The cytotoxicity of AuNPs was low in appropriate incubation time and concentration, and there was no effect on iodine uptake and chemotaxis. The chemotaxis to MDA-MB-231 of BMSCs-Egr1-NIS was identified. AuNPs radiasensitization assay showed that absorbance ( A) 570 nm of MDA-MB-231 cells were significantly deferent in 131I killing groups and blank control group without 131I ( F=60.670, P<0.01), and the cytotoxicity of 131I to MDA-MB-231 cells in the 131I killing groups with 0.20 g/L AuNPs and 0.40 g/L AuNPs ( A570 nm values: 0.87±0.05, 0.41±0.07) was significantly higher than that in the group with 0 g/L AuNPs ( A570 nm=1.39±0.11; both P<0.01). Conclusions:BMSCs, transfected with therapeutic gene Egr1-NIS and incubated with AuNPs, can be used as a carrier to target breast cancer. NIS gene expression of BMSCs-Egr1-NIS was highly promoted in the presence of radioiodine. At the same time, AuNPs can be used as a radiation sensitizer for 131I treatment.
ABSTRACT
ObjectiveNano-graphene oxide quantum dots (GOQDs) can be used to target fluorescent markers. The stem cell labeling is an important method in studying stem cell treatments. Our study aims to explore the possibility of using GOQDs as living cell fluorescent marker materials for human periodontal ligament stem cells (hPDLSCs), and to evaluate the biosecurity and effect as live cell fluorescence markers of GOQDs.Methods GOQDs were testified by TEM, DLS, UV-vis, and PL spectra. hPDLSCs were obtained by tissue cultivation and separated by single cell-derived colony selection. Then the source of the cells was carried out by immunocytochemical staining of anti-vimentin, anti-cytokeratin, and multipotent differentiation was used in the identification of stem cells. hPDLSCs were incubated with different concentrations of GOODs (0, 10, 25, and 50 μg/mL) for 24h and 72 h. Cytotoxicity and proliferation effects were determined using CCK-8, and cell cycles were detected using flow cytometry after the co-culture of GOQDs and hPDLSCs. The fluorescent labeling effect of GOQDs was tested using laser scanning confocal microscopy.ResultsThe characterization of GOQDs showed that the nanoparticles were evenly dispersed in water and showing blue light at 365 nm. TEM and DLS showed GOQDs had good dispersion, and the particle size was (6.36±1.41) nm. Immunocytochemical staining of anti-vimentin was positive while anti-cytokeratin was negative. The results of cytotoxicity showed there were no significant differences in cell activity after incubated with different concentrations of GOODs (0, 5, 10, 25, 50, 100, 200, and 400 μg/mL) (P>0.05), and there was no significant decrease in cell activity between 24h and 72h (P>0.05). There was no significant difference in the proportional distribution of G1, G2, and S phases between the two concentrations of GOQDs (0 μg/mL and 50 μg/mL) (P>0.05). Fluorescent images showed that GOQDs could enter the cell membrane and increase the fluorescence intensity at the concertation of 50 μg/mL.ConclusionGOQDs were confirmed to have good biocompatibility and could be used for live cell labeling of hPDLSCs.
ABSTRACT
ObjectiveNano-graphene oxide quantum dots (GOQDs) can be used to target fluorescent markers. The stem cell labeling is an important method in studying stem cell treatments. Our study aims to explore the possibility of using GOQDs as living cell fluorescent marker materials for human periodontal ligament stem cells (hPDLSCs), and to evaluate the biosecurity and effect as live cell fluorescence markers of GOQDs.Methods GOQDs were testified by TEM, DLS, UV-vis, and PL spectra. hPDLSCs were obtained by tissue cultivation and separated by single cell-derived colony selection. Then the source of the cells was carried out by immunocytochemical staining of anti-vimentin, anti-cytokeratin, and multipotent differentiation was used in the identification of stem cells. hPDLSCs were incubated with different concentrations of GOODs (0, 10, 25, and 50 μg/mL) for 24h and 72 h. Cytotoxicity and proliferation effects were determined using CCK-8, and cell cycles were detected using flow cytometry after the co-culture of GOQDs and hPDLSCs. The fluorescent labeling effect of GOQDs was tested using laser scanning confocal microscopy.ResultsThe characterization of GOQDs showed that the nanoparticles were evenly dispersed in water and showing blue light at 365 nm. TEM and DLS showed GOQDs had good dispersion, and the particle size was (6.36±1.41) nm. Immunocytochemical staining of anti-vimentin was positive while anti-cytokeratin was negative. The results of cytotoxicity showed there were no significant differences in cell activity after incubated with different concentrations of GOODs (0, 5, 10, 25, 50, 100, 200, and 400 μg/mL) (P>0.05), and there was no significant decrease in cell activity between 24h and 72h (P>0.05). There was no significant difference in the proportional distribution of G1, G2, and S phases between the two concentrations of GOQDs (0 μg/mL and 50 μg/mL) (P>0.05). Fluorescent images showed that GOQDs could enter the cell membrane and increase the fluorescence intensity at the concertation of 50 μg/mL.ConclusionGOQDs were confirmed to have good biocompatibility and could be used for live cell labeling of hPDLSCs.
ABSTRACT
Tripterygium wilfordii multiglycoside( GTW),an extract derived from T. wilfordii,has been used for rheumatoid arthritis and other immune diseases in China. However its potential hepatotoxicity has not been investigated completely. Firstly,the content of triptolid( TP) in GTW was 0. 008% confirmed by a LC method. Then after oral administration of GTW( 100,150 mg·kg-1) and TP( 12 μg·kg-1) in female Wistar rats for 24 h,it was found that 150 mg·kg-1 GTW showed more serious acute liver injury than 12 μg·kg-1 TP,with the significantly increased lever of serum ALT,AST,TBA,TBi L,TG and bile duct hyperplasia even hepatocyte apoptosis. The expression of mRNA and proteins of liver bile acid transporters such as BSEP,MRP2,NTCP and OATP were down-regulated significantly by GTW to inhibit bile acid excretion and absorption,resulting in cholestatic liver injury. Moreover,GTW was considered to be involved in hepatic oxidative stress injury,although it down-regulated SOD1 and GPX-1 mRNA expression without significant difference in MDA and GSH levels. In vitro,we found that TP was the main toxic component in GTW,which could inhibit cell viability up to 80% in Hep G2 and LO2 cells at the dose of 0. 1 μmol·L-1. Next a LC-MS/MS method was used to detect the concentration of triptolid in plasma from rats,interestingly,we found that the content of TP in GTW was always higher than in the same amount of TP,suggesting the other components in GTW may affect the TP metabolism. Finally,we screened the substrate of p-glycoprotein( p-gp) in Caco-2 cells treated with components except TP extrated from GTW,finding that wilforgine,wilforine and wilfordine was the substrate of p-gp. Thus,we speculated that wilforgine,wilforine and wilfordine may competitively inhibit the excretion of TP to bile through p-gp,leading to the enhanced hepatotoxity caused by GTW than the same amount of TP.
Subject(s)
Animals , Female , Humans , Rats , Caco-2 Cells , Chemical and Drug Induced Liver Injury , Pathology , Chromatography, Liquid , Diterpenes , Toxicity , Drugs, Chinese Herbal , Toxicity , Epoxy Compounds , Toxicity , Glycosides , Toxicity , Liver , Phenanthrenes , Toxicity , Plant Extracts , Toxicity , Rats, Wistar , Tandem Mass Spectrometry , Tripterygium , ToxicityABSTRACT
Objective Fluorescence-labeled pH-low insertion peptide ( pHLIP ) imaging was used to analyze in vitro acidophilic characteristic of pHLIP and its dynamic distribution in the transplanted breast tumor models. Methods The red fluorescent dye Rhodamine B ( B-pHLIP ) and near-infrared fluorescent dye Cy5 ( Cy5-pHLIP ) were respectively labeled at the hydroxyl terminal of pHLIP for imaging in vitro and in vivo. The fluorescent intensity in the MDA-MB-231 breast cancer cells and the cell vitality were analyzed under different pH values (7.8, 7.4, 7.0, 6.6). In vivo dynamic fluorescent distribution, fluorescent intensi-ties and tumor/non-tumor ( T/NT) ratios in the regions of interest of tumor and normal organs at different time points (2 h, 24 h, 3 d, 7 d) were observed or calculated, and then fluorescence imaging of the isolated tis-sues was finally performed. One-way analysis of variance and least significant difference t test were used to analyze the data. Results Relative fluorescent intensity of B-pHLIP in the MDA-MB-231 cells at pH 6.6, 7.0, 7.4 and 7.8 were (100.00±9.70)％, (69.90±5.50)％, (19.80±1.40)％ and (0.40±0.04)％, re-spectively. There were significant differences between pH 6. 6 group and other groups ( F=230. 504, t=5. 029-17.669, all P<0.05). pHLIP had no significant toxic effect on MDA-MB-231 cells. After Cy5-pHLIP injection in vivo, the fluorescent intensity of tumor in mice gradually decreased, but the T/NT ratios were stable (3.42±0.27, 3.00±1.23, 3.38±0.62 and 3.51±0.37 at 2 h, 24 h, 3 d and 7 d respectively;F=0.192, P>0. 05) . The ex vivo fluorescence distribution showed that Cy5-pHLIP had a higher uptake in the tumor tissue, and the large intestine also secreted a large amount of pHLIP. Conclusions The affinity be-tween pHLIP and tumor cell membrane is significantly increased in the extracellular acidic microenviron-ment. Cy5-pHLIP enables long-term and visual monitoring the tumor-targeted distribution of pHLIP in vivo. However, the high accumulation of pHLIP in the large intestine increases the interpretation complexity of tumor imaging.
ABSTRACT
OBJECTIVE@#To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro.@*METHODS@#placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4 T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4CD25Foxp3Treg in T lymphocytes was accounted.@*RESULTS@#After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4 T lymphocytes were cultured for 48 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (<0.01).@*CONCLUSIONS@#Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4CD25Foxp3Treg in vitro.
Subject(s)
Female , Humans , Pregnancy , Forkhead Transcription Factors , HLA-G Antigens , Mesenchymal Stem Cells , Placenta , T-Lymphocytes, RegulatoryABSTRACT
Objective To investigate whether QKI protein plays any important role in the process of spermatogene-sis.Constructing GC1-spg cell strain which knocked out QKI by the technology of CRISPR/Cas9,and detecting its effect on the proliferation and differentiation of QKI protein in vitro.Methods The plasmid PX330 was used to construct QKI knockout recombinant plasmid, then transfected it to GC1-spg wild-type cells and selected by puromycin.GC1-spg knock-QKI cell strain was identified by Western blot and gene sequencing; The wild-type and knockout cell strain was cultured normally,then detected the growth curve by cell counting kit(CCK8),and using quantitative PCR to get the changes of meiotic-related gene differentiation.Results QKI knockout GC1-spg cell strain was successfully constructed.Compared with the control group,the growth of QKI knockout cell strain was significantly decreased(P<0.05), and the expression of meiosis related molecular marker gene of c-kit, Mtl5 and Pspa2 was significantly decreased(P<0.05).Conclusions QKI proteins can affect reproductive sper-matogenesis by acting on proliferation and differentiation.
ABSTRACT
Objective To evaluate the one-year clinical outcomes in patients with the vulnerable plaque sealing with drug-eluting stents for the treatment of intermediate coronary stenosis. Methods 327 patients with an-giographically intermediate lesions(diameter stenosis 50%~70%)with the vulnerable plaque which were detected by 64 slice coronary CT were prospectively enrolled. Patients were divided into medical therapy group (n = 160) and sirolimus-eluting stent group group(n=160). The incidences of one-year major adverse cardiovascular events (MACE)was evaluated(cardiac death,myocardial infarction ,revascularization). Results The MACE tended to be lower in the sirolimus-eluting stent group than medical therapy group(3.13%vs. 10%,log-rankχ2=6.62,P=0.01). The incident of cardiac death and myocardial infarction were lower in the sirolimus-eluting stent group than medical therapy group(1.25%vs. 5.63%,log-rankχ2=4.61,P=0.03). Conclusion The treatment of the siroli-mus-eluting stent can reduce MACE for the paitents with the vulnerable plaque of intermediate coronary stenosis than medical therapy only.
ABSTRACT
Objective To evaluate the one-year clinical outcomes in patients with the vulnerable plaque sealing with drug-eluting stents for the treatment of intermediate coronary stenosis. Methods 327 patients with an-giographically intermediate lesions(diameter stenosis 50%~70%)with the vulnerable plaque which were detected by 64 slice coronary CT were prospectively enrolled. Patients were divided into medical therapy group (n = 160) and sirolimus-eluting stent group group(n=160). The incidences of one-year major adverse cardiovascular events (MACE)was evaluated(cardiac death,myocardial infarction ,revascularization). Results The MACE tended to be lower in the sirolimus-eluting stent group than medical therapy group(3.13%vs. 10%,log-rankχ2=6.62,P=0.01). The incident of cardiac death and myocardial infarction were lower in the sirolimus-eluting stent group than medical therapy group(1.25%vs. 5.63%,log-rankχ2=4.61,P=0.03). Conclusion The treatment of the siroli-mus-eluting stent can reduce MACE for the paitents with the vulnerable plaque of intermediate coronary stenosis than medical therapy only.
ABSTRACT
<p><b>OBJECTIVE</b>To study on the expression patterns of proteins associated with cell junctions in the developing mouse testes.</p><p><b>METHOD</b>The expression levels of reproductive related cell lines spermatogonia cell line GC1 spg, spermatocyte cell line GC2 spg, leydig cell line TM3, and sertoli cell line TM4, primary sertoli cells, and 1-6-week mouse testes were analyzed using Western blot.</p><p><b>RESULTS</b>The sertoli cell junction-associated membrane proteins adhesion molecule A, Occludin and Claudin, and the sertoli-germ cell junction-associated membrane proteins junctional adhesion molecule C, Nectin-3, and E-cadherin were stage-specific in the seminiferous tubules in the mouse testes. The adaptor proteins associated with cell juctions zonula occludens-1, zonula occludens-2, Afadin, Β-catenin, and CD2-associated protein were not stage-specific in the seminiferous tubules in the mouse testes.</p><p><b>CONCLUSIONS</b>In the seminiferous tubules in the mouse testes, the membrane proteins associated with cell junctions are stage-specific. However, the expressions of adaptor proteins associated with cell junctions do not obviously change.</p>
Subject(s)
Animals , Humans , Male , Mice , Adaptor Proteins, Signal Transducing , Metabolism , Cdh1 Proteins , Metabolism , Cell Adhesion Molecules , Metabolism , Cell Line , Cytoskeletal Proteins , Metabolism , Intercellular Junctions , Metabolism , Membrane Proteins , Metabolism , Microfilament Proteins , Metabolism , Nectins , Seminiferous Tubules , Cell Biology , Metabolism , Sertoli Cells , Cell Biology , Testis , Cell Biology , Zonula Occludens-1 Protein , Metabolism , Zonula Occludens-2 Protein , Metabolism , beta Catenin , MetabolismABSTRACT
AIM: To investigate the efficiency of intravitreal ranibizumab therapy ( IVR ) for polypoidal choroidal vasculopathy ( PCV) in single or multiple polyps. METHODS: A total 63 patients diagnosed with PCV in Shaoxing City People's Hospital from May 2013 to May 2015 were enrolled and divided into single polyp group and multiple polyps group. All patients received intravitreal ranibizumab 3 monthly and were followed up for 12mo. Observe the changes of best corrected visual acuity ( BCVA ) and central retinal thickness ( CRT ) at different time points. RESULTS: The single polyps group exhibited a better BCVA, shorter greatest linear dimension, and lower prevalence of fibro - vascular pigment epithelial detachment compared with the multiple polyp group before treatment (P CONCLUSION: IVR meet better result in PCV patients with multiple polyp and polyp numbers may be valuable to prognosis.
ABSTRACT
Objective:To observe the effects of different moxibustion times on proteins of transient receptor potential vanilloid 3 (TRPV3) ion channel protein and synovial cell apoptosis in rats with rheumatoid arthritis (RA), to provide a new basis for the anti-inflammatory mechanism of moxibustion. Methods:A total of 50 Sprague-Dawley (SD) rats were divided into a normal group, a model group, moxibustion groupⅠ, moxibustion groupⅡ and moxibustion groupⅢ by complete randomization, with 10 rats in each group. Rats in the normal group were bred routinely, and rats in the model group were also bred routinely after successful modeling. After successful modeling, rats in moxibustion groupⅠ,Ⅱ andⅢ accepted consecutive moxibustion at Zusanli (ST 36) and Shenshu (BL 23) for 15 d, once a day, respectively 5 min, 20 min and 30 min for each session. The degree of paw edema was observed and recorded. Immunohistochemical assay was used to detect the protein expression of TRPV3 ion channel in dorsal root ganglia and spinal cord dorsal horn. Terminal-deoxynucleoitidyl transferase-mediated nick end labeling (TUNEL) was used to detect apoptotic synovial cell number. Results: At the end of treatment, paw circumference of rats in moxibustion groupⅡ andⅢ were significantly reduced as compared with that in the model group (P<0.05). TRPV3 ion channel protein expression of dorsal root ganglia and spinal cord dorsal horn was higher in the model group than that in the normal group (P<0.05); the TRPV3 ion channel protein expressions of dorsal root ganglia and spinal cord dorsal horn in moxibustion groupⅡ andⅢ were higher than that in moxibustion groupⅠ (P<0.05); apoptotic synovial cell number in the model group was larger than that in the normal group (P<0.05), and apoptotic synovial cell numbers in moxibustion groupⅡ andⅢ were significantly higher than that in the model group (P<0.05). Conclusion:Moxibustion of appropriate time could induce TRPV3 expression, and promote synovial cell apoptosis.
ABSTRACT
Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene,and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma.Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro,then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP.pLVX-CMV-0-EGFP was constructed as control.BMSCs were isolated,cultured,and transfected by lentivirus.The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected.The expression of NIS gene was examined by Western blot.Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4.The nude mice intracranial glioma models were established.MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein.Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged.BMSCs were successfully isolated and cultured.Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection.The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP,but not in BMSCs-EGFP.BMSCs-NIS-EGFP showed significantly more uptake of 125I (nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4.The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection.Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma.It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma.
ABSTRACT
AlM:To compare the efficacy and safety of latanoprost and brimonidine in the treatment of open angle glaucoma, and provide reference for rational drug use.METHODS:A total of 121 cases ( 136 eyes ) who were diagnosed as primary open angle glaucoma were selected in this study, and they were randomly divided into experimental group (62 cases, 70 eyes) and control group ( 59 cases, 66 eyes) according to different drug treatment. Patients in the control group received brimonidine eye drops twice a day, while patients in the experimental group received latanoprost eye drops once a day. The intraocular pressure, visual acuity and adverse reactions were checked of the two groups in the following 3mo.RESULTS:The intraocular pressure of patients in the control group was 18. 1 ± 1. 3mmHg, while the experimental group was 17. 0 ± 0. 9mmHg after 12wk of treatment, which were both lower than before (P<0. 05). The fluctuation of intraocular pressure in the experimental group was significantly lower than that of the control group. There was no significant difference in the LogMAR visual acuity between before and after treatment in the control group, while the LogMAR visual acuity of the experimental group was significantly improved. The control group had hyperemia, burning sensation, tearing, eyelid edema and other adverse side effects, and the experimental group had little adverse reactions. CONCLUSlON: Latanoprost can significantly reduce intraocular pressure in glaucoma patients with in the follow- up time, and reduce the impact of elevated intraocular pressure in the vision of glaucoma patients, with little adverse reaction, worthy of clinical application.
ABSTRACT
Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P＜0.05).Furthermore,a notable decreased cell survival rate (38.3％) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8％ in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2％ and 10.9％ respectively,F=19.926,45.409;both P＜0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.
ABSTRACT
Objective To construct a recombinant lentivirus vector containing the human NIS gene and HIF-1α with the myosin light chain-2v(MLC-2v) as a promoter and to investigate the specific expression and feasibility of NIS as a reporter gene in cardiomyocytes.Methods The target gene HIF-1α and NIS were subcloned into the lentivirus (Lv)-elongation factor (EF)1-HIF-1α-internal ribosome entry site (IRES)-NIS and Lv-MLC-HIF-1α-IRES-NIS lentivirus vectors.The recombinated vectors were transfected into Hela cells by lipofectamine 2000.The expression of HIF-1α and NIS in the transfected Hela cells was detected by indirect immunofluorescence and Western blot.The H9C2 cells were exposed to different multiplicities of infection (MOI; 5,10,20,40) with packaged virus particles.The infection efficiency was detected by Western blot.MOI 20 was used for H9C2,NIH-3T3 and L6 cell lines and the specificity of the MLC-2v promoter was detected by the count of NIS protein in the 3 different cell lines with Western blot.The function and features of NIS protein were evaluated by dynamic iodine uptake and NaClO4 iodine uptake inhibition tests in vitro.Two-sample t test was used to analyze the data.Results The two recombinant lentivirus vectors were constructed successfully.The HIF-1α protein was expressed in the cytoplasm and the NIS protein was expressed on the cell membrane in Hela cells.The grey levels of NIS and HIF-1α proteins in the positive control were 69.8 and 71.9,respectively,which were 109.4 and 92.7 after being prompted by EF1,and 141.9 and 132.4 by MLC-2v.The expression of these proteins was much higher by EF1 promoter than that by MLC-2v promoter.The optimal MOI for the Lv-MLC-HIF-1α-IRES-NIS virus to infect H9C2 cells was 20.With the MOI of 20,the grey levels of NIS protein promoted by EF1 were 23.4,29.8 and 28.6 for H9C2,NIH-3T3 and L6 cells infected with Lv-EF1-HIF-1α-IRES-NIS virus,respectively.The expression of NIS protein promoted by MLC-2v was much higher in H9C2 cells than the other two cell lines.The grey level of NIS protein was 157.9 in H9C2 cells,178.8 in L6 cells and 217.3 in NIH-3T3 cells.The NIS protein expressed in infected H9C2 cells showed high radioiodine uptake.The peak of iodine uptake was 4 287.2 counts · min-1 at 40 min which was 16.85 times of the control group (254.4 counts · min-1) (t=5.34,P＜ 0.01).The inhibition rate of iodine uptake was up to 85.5％ (3 666.4/4 287.2,t=21.3,P＜0.01) by NaClO4.Conclusions MLC-2v promoter allows specific expression of the external gene HIF-1α and NIS in myocardium.The cardiomyocytes transfected with NIS gene acquires the function of iodine uptake.Therefore,NIS may have a potential to be the reporter gene to monitor the external gene therapy in ischemic cardiomyopathy.