Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 24
Filter
1.
Eur J Anaesthesiol ; 41(3): 226-233, 2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38230449

ABSTRACT

BACKGROUND: Sleep disturbances in the peri-operative period have been associated with adverse outcomes, including postoperative delirium (POD). However, research on sleep quality during the immediate postoperative period is limited. OBJECTIVES: This study aimed to investigate the association between sleep quality on the night of the operative day assessed using the Sleep Quality Numeric Rating Scale (SQ-NRS), and the incidence of POD in a large cohort of surgical patients. DESIGN: A prospective cohort study. SETTING: A tertiary hospital in China. PATIENTS: This study enrolled patients aged 65 years or older undergoing elective surgery under general anaesthesia. The participants were categorised into the sleep disturbance and no sleep disturbance groups according to their operative night SQ-NRS. MAIN OUTCOME MEASURES: The primary outcome was delirium incidence, whereas the secondary outcomes included acute kidney injury, stroke, pulmonary infection, cardiovascular complications and all-cause mortality within 1 year postoperatively. RESULTS: In total, 3072 patients were included in the analysis of this study. Among them, 791 (25.72%) experienced sleep disturbances on the night of operative day. Patients in the sleep disturbance group had a significantly higher risk of developing POD (adjusted OR 1.43, 95% CI 1.11 to 1.82, P  = 0.005). Subgroup analysis revealed that age 65-75 years; male sex; ASA III and IV; haemoglobin more than 12 g l -1 ; intra-operative hypotension; surgical duration more than 120 min; and education 9 years or less were significantly associated with POD. No interaction was observed between the subgroups. No significant differences were observed in the secondary outcomes, such as acute kidney injury, stroke, pulmonary infection, cardiovascular complications and all-cause mortality within 1 year postoperatively. CONCLUSIONS: The poor subjective sleep quality on the night of operative day was independently associated with increased POD risk, especially in certain subpopulations. Optimising peri-operative sleep may reduce POD. Further research should investigate potential mechanisms and causal relationships. TRIAL REGISTRY: chictr.org.cn: ChiCTR1900028545.


Subject(s)
Acute Kidney Injury , Cardiovascular Infections , Delirium , Emergence Delirium , Stroke , Aged , Humans , Male , Cardiovascular Infections/complications , Delirium/diagnosis , Delirium/epidemiology , Delirium/etiology , Emergence Delirium/diagnosis , Emergence Delirium/epidemiology , Emergence Delirium/etiology , Postoperative Complications/diagnosis , Postoperative Complications/epidemiology , Postoperative Complications/etiology , Prospective Studies , Risk Factors , Sleep Quality , Female
2.
J Am Heart Assoc ; 12(24): e029745, 2023 Dec 19.
Article in English | MEDLINE | ID: mdl-38084712

ABSTRACT

BACKGROUND: Cardiac hypertrophy (CH) is a well-established risk factor for many cardiovascular diseases and a primary cause of mortality and morbidity among older adults. Currently, no pharmacological interventions have been specifically tailored to treat CH. OTUD7B (ovarian tumor domain-containing 7B) is a member of the ovarian tumor-related protease (OTU) family that regulates many important cell signaling pathways. However, the role of OTUD7B in the development of CH is unclear. Therefore, we investigated the role of OTUD7B in CH. METHODS AND RESULTS: OTUD7B knockout mice were used to assay the role of OTUD7B in CH after transverse aortic coarctation surgery. We further assayed the specific functions of OTUD7B in isolated neonatal rat cardiomyocytes. We found that OTUD7B expression decreased in hypertrophic mice hearts and phenylephrine-stimulated neonatal rat cardiomyocytes. Furthermore, OTUD7B deficiency exacerbated transverse aortic coarctation surgery-induced myocardial hypertrophy, abnormal cardiac function, and fibrosis. In cardiac myocytes, OTUD7B knockdown promoted phenylephrine stimulation-induced myocardial hypertrophy, whereas OTUD7B overexpression had the opposite effect. An immunoprecipitation-mass spectrometry analysis showed that OTUD7B directly binds to KLF4 (Krüppel-like factor 4). Additional molecular experiments showed that OTUD7B impedes KLF4 degradation by inhibiting lysine residue at 48 site-linked ubiquitination and suppressing myocardial hypertrophy by activating the serine/threonine kinase pathway. CONCLUSIONS: These results demonstrate that the OTUD7B-KLF4 axis is a novel molecular target for CH treatment.


Subject(s)
Aortic Coarctation , Kruppel-Like Factor 4 , Mice , Rats , Animals , Cardiomegaly/genetics , Cardiomegaly/prevention & control , Cardiomegaly/metabolism , Phenylephrine/pharmacology , Phenylephrine/metabolism , Mice, Knockout , Ubiquitination , Myocytes, Cardiac/metabolism , Mice, Inbred C57BL , Endopeptidases/metabolism , Endopeptidases/pharmacology
3.
Adv Sci (Weinh) ; 10(33): e2303639, 2023 Nov.
Article in English | MEDLINE | ID: mdl-37807820

ABSTRACT

To explore carbide superconductors with higher transition temperature, two novel carbon structures of cage-network are designed and their superconductivity is studied by doping metals. MC6 and MC10 are respectively identified as C24 and C32 cage-network structures. This study finds that both carbon structures drive strong electron-phonon interaction and can exhibit superconductivity above liquid nitrogen temperature. Importantly, the superconducting transition temperatures above 100 K are predicted to be achieved in C24 -cage-network systems doped by Na, Mg, Al, In, and Tl at ambient pressure, which is far higher than those in graphite, fullerene, and other carbides. Meanwhile, the superconductivity of cage-network carbides is also found to be sensitive to the electronegativity and concentration of dopant M. The result indicates that the higher transition temperatures can be obtained by optimizing the carbon-cage-network structures and the doping conditions. The study suggests that the carbon-cage-network structure is a direction to explore high-temperature superconducting carbides.

4.
Nanoscale Res Lett ; 17(1): 102, 2022 Oct 27.
Article in English | MEDLINE | ID: mdl-36301360

ABSTRACT

Light-stimulated synaptic devices are promising candidates for the development of artificial intelligence systems because of their unique properties, which include broad bandwidths, low power consumption, and superior parallelism. The key to develop such devices is the realization of photoelectric synaptic behavior in them. In this work, visible-light-stimulated synaptic transistors based on CdSe quantum dot (CdSe QD)/amorphous In-Ga-Zn-O hybrid channels are proposed. This design can not only improve the charge separation efficiency of the photogenerated carriers, but also can induce delayed decay of the photocurrent. The improved charge separation efficiency enhances the photoelectric properties significantly, while the delayed decay of the photocurrent led to the realization of photoelectric synaptic behaviors. This simple and efficient method of fabricating light-stimulated phototransistors may inspire new research progress into the development of artificial intelligence systems.

5.
Inorg Chem ; 61(13): 5318-5325, 2022 Apr 04.
Article in English | MEDLINE | ID: mdl-35302364

ABSTRACT

Herein, a N-rich metal-organic framework (MOF) with four kinds of cages, Zn4(ade)2(TCA)2(H2O) (NENU-1000, Hade = adenine, H3TCA = 4,4',4″-tricarboxytriphenylamine, NENU = Northeast Normal University), was prepared by the mixed-ligand strategy. Cationic dyes can be selectively absorbed by NENU-1000 at proper concentrations, but not neutral and anionic dyes, which perhaps can be assigned to the N-rich neutral framework of NENU-1000. When NENU-1000 was introduced to a relatively lower concentration of cationic dye solutions (e.g., rhodamine B or basic red 2), the colors of these systems faded quickly. Furthermore, the faded solutions can be used for the detection of methanol and other small alcohol molecules with either the naked eye or common UV-vis spectra. The effect of the length of carbon chain, the position of the -OH group, and the number of the hydroxyl group of the alcohols was explored for the color development rate. In addition, the performance of NENU-1000 in iodine sorption and release was also studied.


Subject(s)
Coloring Agents , Metal-Organic Frameworks , Alcohols , Carbon , Humans
6.
Sci Rep ; 9(1): 13609, 2019 09 20.
Article in English | MEDLINE | ID: mdl-31541168

ABSTRACT

Assortative matching (AM) can be theoretically an effective means to facilitate cooperation. We designed a controlled lab experiment with three treatments on multi-round prisoner's dilemma. With matching based on weighted history (WH) as surrogate for AM, we show that adding pro-social dummies to the WH treatment may significantly improve cooperation, compared to both the random matching and the WH treatment. In society where assortative matching is effective and promoted by the underlying culture, institutional promotion of virtue role models can be interpreted as generating additional pro-social dummies, so as to move the initial state of cooperators into the basin of attraction for a highly cooperative polymorphic equilibrium.

7.
Proc Natl Acad Sci U S A ; 115(40): 9968-9973, 2018 10 02.
Article in English | MEDLINE | ID: mdl-30224497

ABSTRACT

Sustaining cooperation in social dilemmas is a fundamental objective in the social and biological sciences. Although providing a punishment option to community members in the public goods game (PGG) has been shown to effectively promote cooperation, this has some serious disadvantages; these include destruction of a society's physical resources as well as its overall social capital. A more efficient approach may be to instead employ a reward mechanism. We propose an endogenous reward mechanism that taxes the gross income of each round's PGG play and assigns the amount to a fund; each player then decides how to distribute his or her share of the fund as rewards to other members of the community. Our mechanism successfully reverses the decay trend and achieves a high level of contribution with budget-balanced rewards that require no external funding, an important condition for practical implementation. Simulations based on type-specific estimations indicate that the payoff-based conditional cooperation model explains the observed treatment effects well.


Subject(s)
Cooperative Behavior , Models, Economic , Reward , Humans
8.
J Phys Condens Matter ; 30(24): 245703, 2018 Jun 20.
Article in English | MEDLINE | ID: mdl-29749961

ABSTRACT

Light-element compounds hold great promise of high critical temperature superconductivity judging from the theoretical perspective. A hydrogen-rich material, benzene, is such a kind of candidate but also an organic compound. A series of first-principles calculations are performed on the electronic structures, dynamics properties, and electron-phonon interactions of solid benzene at high pressures. Benzene is found to be dynamically stable in the pressure range of 180-200 GPa and to exhibit superconductivity with a maximum transition temperature of 20 K at 195 GPa. The phonon modes of carbon atoms are identified to mainly contribute to the electron-phonon interactions driving this superconductivity. The predicted superconductivity in this simplest pristine hydrocarbon shows a common feature in aromatic hydrocarbons and also makes it a bridge to organic and hydrogen-rich superconductors.

9.
Vet Microbiol ; 183: 85-91, 2016 Feb 01.
Article in English | MEDLINE | ID: mdl-26790939

ABSTRACT

Pigs are susceptible to both human and avian influenza viruses and therefore have been proposed to be mixing vessels for the generation of pandemic influenza viruses through reassortment. In this study, for the first time, we report the isolation and genetic analyses of three novel triple-reassortant H1N1 swine influenza viruses from pigs in Tianjin, Northern China. Phylogenetic analysis showed that these novel viruses contained genes from the 2009 pandemic H1N1 (PB2, PB1, PA and NP), Eurasian swine (HA, NA and M) and triple-reassortant swine (NS) lineages. This indicated that the reassortment among the 2009 pandemic H1N1, Eurasian swine and triple-reassortant swine influenza viruses had taken place in pigs in Tianjin and resulted in the generation of new viruses. Furthermore, three human-like H1N1, two classical swine H1N1 and two Eurasian swine H1N1 viruses were also isolated during the swine influenza virus surveillance from 2009 to 2013, which indicated that multiple genetic lineages of swine H1N1 viruses were co-circulating in the swine population in Tianjin, China. The emergence of novel triple-reassortant H1N1 swine influenza viruses may be a potential threat to human health and emphasizes the importance of further continuous surveillance.


Subject(s)
Influenza A Virus, H1N1 Subtype/isolation & purification , Orthomyxoviridae Infections/veterinary , Reassortant Viruses/isolation & purification , Swine Diseases/virology , Animals , China , Genes, Viral/genetics , Influenza A Virus, H1N1 Subtype/classification , Influenza A Virus, H1N1 Subtype/genetics , Orthomyxoviridae Infections/virology , Phylogeny , RNA, Viral/genetics , Reassortant Viruses/classification , Reassortant Viruses/genetics , Swine
10.
J Zhejiang Univ Sci B ; 16(8): 709-19, 2015 Aug.
Article in English | MEDLINE | ID: mdl-26238546

ABSTRACT

The presence of yeast cells could stimulate hydrogen utilization of acetogens and enhance acetogenesis. To understand the roles of acetogens in rumen fermentation, an in vitro rumen fermentation experiment was conducted with addition of acetogen strain (TWA4) and/or Saccharomyces cerevisiae fermentation product (XP). A 2×2 factorial design with two levels of TWA4 (0 or 2×10(7) cells/ml) and XP (0 or 2 g/L) was performed. Volatile fatty acids (VFAs) were increased (P<0.05) in XP and TWA4XP, while methane was increased only in TWA4XP (P<0.05). The increase rate of microorganisms with formyltetrahydrofolate synthetase, especially acetogens, was higher than that of methanogens under all treatments. Lachnospiraceae was predominant in all acetogen communities, but without close acetyl-CoA synthase (ACS) amino acid sequences from cultured isolates. Low-Acetitomaculum ruminis-like ACS was predominant in all acetogen communities, while four unique phylotypes in XP treatment were all amino acid identified low-Eubacterium limosum-like acetogens. It differs to XP treatment that more low-A. ruminis-like and less low-E. limosum-like sequences were identified in TWA4 and TWA4XP treatments. Enhancing acetogenesis by supplementation with an acetogen strain and/or yeast cells may be an approach to mitigate methane, by targeting proper acetogens such as uncultured low-E. limosum-like acetogens.


Subject(s)
Acetates/metabolism , Fermentation/physiology , Microbial Consortia/physiology , Rumen/microbiology , Rumen/physiology , Saccharomyces cerevisiae/physiology , Animals , Cattle , Species Specificity
11.
Asian Pac J Cancer Prev ; 16(7): 3035-42, 2015.
Article in English | MEDLINE | ID: mdl-25854402

ABSTRACT

BACKGROUND: Isorhamnetin (Iso), a novel and essential monomer derived from total flavones of Hippophae rhamnoides that has long been used as a traditional Chinese medicine for angina pectoris and acute myocardial infarction, has also shown a spectrum of antitumor activity. However, little is known about the mechanisms of action Iso on cancer cells. OBJECTIVES: To investigate the effects of Iso on A549 lung cancer cells and underlying mechanisms. MATERIALS AND METHODS: A549 cells were treated with 10~320 µg/ml Iso. Their morphological and cellular characteristics were assessed by light and electronic microscopy. Growth inhibition was analyzed by MTT, clonogenic and growth curve assays. Apoptotic characteristics of cells were determined by flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay, immunocytochemistry and terminal deoxynucleotidyl transferase nick end labeling (TUNEL) . Tumor models were setup by transplanting Lewis lung carcinoma cells into C57BL/6 mice, and the weights and sizes of tumors were measured. RESULTS: Iso markedly inhibited the growth of A549 cells with induction of apoptotic changes. Iso at 20 µg/ml, could induce A549 cell apoptosis, up-regulate the expression of apoptosis genes Bax, Caspase-3 and P53, and down-regulate the expression of Bcl-2, cyclinD1 and PCNA protein. The tumors in tumor-bearing mice treated with Iso were significantly smaller than in the control group. The results of apoptosis-related genes, PCNA, cyclinD1 and other protein expression levels of transplanted Lewis cells were the same as those of A549 cells in vitro. CONCLUSIONS: Iso, a natural single compound isolated from total flavones, has antiproliferative activity against lung cancer in vitro and in vivo. Its mechanisms of action may involve apoptosis of cells induced by down-regulation of oncogenes and up-regulation of apoptotic genes.


Subject(s)
Cell Proliferation/drug effects , Lung Neoplasms/drug therapy , Quercetin/analogs & derivatives , Animals , Apoptosis/drug effects , Carcinoma, Lewis Lung/diet therapy , Carcinoma, Lewis Lung/metabolism , Caspase 3/metabolism , Cell Line, Tumor , Cyclin D1/metabolism , Down-Regulation/drug effects , Humans , Lung Neoplasms/metabolism , Mice , Mice, Inbred C57BL , Proliferating Cell Nuclear Antigen/metabolism , Quercetin/pharmacology , Tumor Suppressor Protein p53/metabolism , Up-Regulation/drug effects , bcl-2-Associated X Protein/metabolism
12.
World J Gastroenterol ; 18(8): 754-66, 2012 Feb 28.
Article in English | MEDLINE | ID: mdl-22371635

ABSTRACT

AIM: To investigate the effects of resistin-like molecule ß (RELMß) over-expression on the invasion, metastasis and angiogenesis of gastric cancer cells. METHODS: Human RELMß encoding expression vector was constructed and transfected into the RELMß lowly-expressed gastric cancer cell lines SGC-7901 and MKN-45. Gene expression was measured by Western blotting, reverse transcription polymerase chain reaction (PCR) and real-time quantitative PCR. Cell proliferation was measured by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetry, colony formation and 5-ethynyl-20-deoxyuridine incorporation assays. The in vitro migration, invasion and metastasis of cancer cells were measured by cell adhesion assay, scratch assay and matrigel invasion assay. The angiogenic capabilities of cancer cells were measured by tube formation of endothelial cells. RESULTS: Transfection of RELMß vector into SGC-7901 and MKN-45 cells resulted in over-expression of RELMß, which did not influence the cellular proliferation. However, over-expression of RELMß suppressed the in vitro adhesion, invasion and metastasis of cancer cells, accompanied by decreased expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. Moreover, transfection of RELMß attenuated the expression of vascular endothelial growth factor and in vitro angiogenic capabilities of cancer cells. CONCLUSION: Over-expression of RELMß abolishes the invasion, metastasis and angiogenesis of gastric cancer cells in vitro, suggesting its potentials as a novel therapeutic target for gastric cancer.


Subject(s)
Intercellular Signaling Peptides and Proteins/metabolism , Neoplasm Metastasis/pathology , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Animals , Cell Adhesion/physiology , Cell Line, Tumor , Cell Movement/physiology , Cell Proliferation , Humans , Intercellular Signaling Peptides and Proteins/genetics , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/genetics , Matrix Metalloproteinase 9/metabolism , Neovascularization, Pathologic , Proto-Oncogene Protein c-ets-1/genetics , Proto-Oncogene Protein c-ets-1/metabolism , Stomach Neoplasms/genetics , Transfection , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/metabolism
13.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 41(3): 382-5, 2010 May.
Article in Chinese | MEDLINE | ID: mdl-20629303

ABSTRACT

OBJECTIVE: To study the effect of shRNA expressing vector targeting to id1 gene on the biological behavior of the esophageal cancer cell. METHODS: The specific shRNA sequence was designed, synthesized and cloned into pGFU6/neo vector. Then, the resulting recombinants were transfected into Eca-109 cells using Lipofetamine 2000 following the instruction of the manufactures. The expression levels of id1 mRNA were analyzed by RT-PCR and the levels of Id1, p16 and PCNA protein were detected by Western blot assay. The growth of Eca-109 cells was analyzed using clone formation assay and trypan blue exclusion assay. Distribution of cell cycle and proliferation of Eca-109 cells were assessed by flow cytometry. RESULTS: The sequence of recombinant plasmid was confirmed by sequence analysis. The expression levels of id1 mRNA and Id1 protein significantly decreased in Eca-109 cells transfected with pGFU6/neo-Id1-1 vector compared to the negative control group (P < 0.05). The recombinant plasmid expressing id1 shRNA could effectively inhibit cell proliferation and induce cell cycle arrest in G0/G1 phase with a remarkably decrease of cells in S phase (P < 0.05). CONCLUSION: All the above data suggested that the expression of id1 gene was significantly inhibited in Eca-109 cells transfected with pGFU6/neo-id1-1 recombinant. Furthermore, the proliferation of id1 knockdown cells was depressed.


Subject(s)
Esophageal Neoplasms/genetics , Esophageal Neoplasms/pathology , Inhibitor of Differentiation Protein 1/genetics , RNA, Small Interfering/genetics , Cell Line, Tumor , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p16 , Humans , Inhibitor of Differentiation Protein 1/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Proliferating Cell Nuclear Antigen/genetics , Proliferating Cell Nuclear Antigen/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Transfection
14.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 38(4): 583-6, 598, 2007 Jul.
Article in Chinese | MEDLINE | ID: mdl-17718416

ABSTRACT

OBJECTIVE: To develop DNA vaccine for Legionella pneumophila. METHODS: PAL gene of Legionella pneumophila was amplified with PCR. The amplified DNA was ligated to pcDNA3.1(+) vector. The recombinant plasmid, which was identified by restriction analysis, PCR and sequence analysis, was named pcDNA3.1-PAL. The NIH3T3 cells were transfected with the recombinant plasmid pcDNA3.1-PAL by Lipofection. Transient and stable expression products of the PAL gene were detected by immunofluorescence and RT-PCR. RESULTS: The recombinant plasmid pcDNA3.1-PAL expressed PAL protein in the eukaryotic cell NIH3T3. CONCLUSION: This study has built a foundation for the development of PAL gene DNA vaccine for Legionella pneumophila.


Subject(s)
Legionella pneumophila/genetics , Lipoproteins/genetics , Animals , Cloning, Molecular , Gene Expression , Genetic Engineering , Immunohistochemistry , Legionella pneumophila/immunology , Lipoproteins/isolation & purification , Mice , NIH 3T3 Cells , Plasmids/genetics , Reverse Transcriptase Polymerase Chain Reaction , Vaccines, DNA/genetics , Vaccines, DNA/immunology
15.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 38(2): 306-8, 2007 Mar.
Article in Chinese | MEDLINE | ID: mdl-17441356

ABSTRACT

OBJECTIVE: To explore the relationship between Aurora A expression and cell sensitivity to taxol, and elucidate the effect of Aurora A on the treatment efficacy of taxol for prostate cancer. METHODS: A cell clone highly expressing Aurora A was constructed through transfected with full length Aurora A gene, and then termed as Du145-Aurora A. Cells were treated with different dose taxols. The growth inhibition rates of cells were detected by MTT assay, and the cell apoptosis rates were detected through flow cytometry. The growth inhibition and apoptosis rates of Du145-Aurora A and Du145 cells were compared for Aurora A up-regulated expression. Simultaneously, cells Du145-Aurora A were treated with DNAzymes for down-regulating the Aurora A expression, and then the variations of cell growth inhibited rate and apoptosis rate were detected. RESULTS: Up-regulation of Aurora A expression reduced the cell inhibition and apoptosis rates when treated by taxol, but the down-regulation of Aurora A expression after DNAzymes applied increased the cell inhibition and apoptosis rates when treated by taxol. CONCLUSION: There is a positive relationship between Aurora A expression and treatment efficacy of taxol for prostate cancer, and inhibiting Aurora A expression will enhance the treatment efficacy of taxol for prostate cancer.


Subject(s)
Antineoplastic Agents/pharmacology , Gene Expression Regulation, Neoplastic , Paclitaxel/pharmacology , Prostatic Neoplasms/drug therapy , Prostatic Neoplasms/pathology , Protein Serine-Threonine Kinases/metabolism , Antineoplastic Agents/therapeutic use , Aurora Kinases , Cell Line, Tumor , Cell Proliferation/drug effects , Down-Regulation , Humans , Male , Paclitaxel/therapeutic use , Prostatic Neoplasms/genetics , Up-Regulation
16.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 36(6): 773-5, 2005 Nov.
Article in Chinese | MEDLINE | ID: mdl-16334549

ABSTRACT

OBJECTIVE: To construct recombinant plasmid of Legionella pneumophila mip gene and detect its expression in NIH3T3 cells. METHODS: mip gene of Legionella pneumophila was amplified by PCR. The amplified DNA was ligated to pcDNA3.1(+) vector. The recombinant plasmid was named pcDNA3.1-mip. NIH3T3 cell was transfected by recombinant plasmid pcDNA3.1-mip with Lipofection strategy. Transient and stable products of mip gene were detected by immunofluorescence and Western-blot. RESULTS: It was found that there was high green fluorescence on the cell membrane and inside the cell. It showed that NIH3T3 cell was transfected by pcDNA3.1-mip successfully. Rabbit serum antibody of Legionella pneumophila detected the NIH3T3 cell transfected with pcDNA3.1-mip. There was the protein in relative molecular weight 24 X 10(3), whereas no evidence for the protein in NIH3T3 cell transfected with pcDNA3.1(+) was seen. The protein expression of mip gene was shown. CONCLUSION: We have successfully constructed the recombinant plasmid of Legionella pneumophila mip gene and detected the relative molecular weight 24 X 10(3) Mip protein in NIH3T3 cells.


Subject(s)
Immunophilins/biosynthesis , Legionella pneumophila/genetics , Membrane Proteins/biosynthesis , Peptidylprolyl Isomerase/biosynthesis , 3T3 Cells , Animals , Bacterial Proteins , Cloning, Molecular , Eukaryotic Cells/metabolism , Genetic Vectors , Humans , Immunophilins/genetics , Membrane Proteins/genetics , Mice , Peptidylprolyl Isomerase/genetics , Plasmids/genetics , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombination, Genetic , Transfection
17.
Space Med Med Eng (Beijing) ; 18(3): 222-3, 2005 Jun.
Article in Chinese | MEDLINE | ID: mdl-16161280

ABSTRACT

OBJECTIVE: To investigate the role of circadian gene mPeriod2 (mPer2) on tumor proliferation and apoptosis. METHOD: The eukaryotic mPer2 expression vector (pcDNA 3.1-mPer2) based on pcDNA 3.1 was transfected into the cultured Lewis tumor cell by liposome method in vivo. The expression of mPer2 in transfected Lewis tumor cell was detected by immunohistochemistry and flowcytometry. The stable transfected cell line was screened by G418 and its proliferation and apoptosis was examined by flowcytometry. RESULT: Expression of PERIOD2 protein in pcDNA 3.1-mPer2 transfected Lewis tumor cell was proved by immunohistochemistry and flowcytometry. Flowcytometry result of mPer2 expression Lewis tumor cell showed less proliferation and high rate of apoptosis. CONCLUSION: mPer2 expression can suppress the proliferation of tumor cell and increase the apoptosis of it.


Subject(s)
Circadian Rhythm/genetics , Nuclear Proteins/metabolism , Transcription Factors/metabolism , Animals , Apoptosis , Carcinoma, Lewis Lung/genetics , Cell Cycle Proteins , Cell Division , DNA, Complementary/genetics , Flow Cytometry , Gene Expression , Mice , Nuclear Proteins/genetics , Period Circadian Proteins , Transcription Factors/genetics , Transfection , Tumor Cells, Cultured/immunology
18.
Acta Biochim Biophys Sin (Shanghai) ; 37(3): 199-204, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15756423

ABSTRACT

The mip gene of Legionella pneumophila and the ctxB gene of Vibrio cholerae were amplified by PCR respectively. The amplified cDNA was ligated to the pcDNA3.1(+) vector. The recombinant plasmids pcDNA3.1-mip and pcDNA3.1-ctxB were identified by restriction analysis and PCR, and further confirmed by sequencing analysis. NIH3T3 cells were transfected with pcDNA3.1-mip and pcDNA3.1-ctxB according to the Lipofection method. Transient and stable products of the co-expression of the mip gene and ctxB gene were detected by immunofluorescence and Western blotting. The results showed that NIH3T3 cells were successfully transfected, and that the transiently and stably co-expressed products can be detected in the transfected cells. To detect the humoral and cellular immune response in immunized mice induced by the co-mmunization of the mip and ctxB genes, female BALB/c mice were immunized intramuscularly with pcDNA3.1-mip and pcDNA3.1-ctxB. The results showed that the specific antibody titer and the cytotoxic T-lymphocyte response for pcDNA3.1-mip immunization and co-immunization were increased compared with that of pcDNA3.1(+) immunization. Furthermore, the specific antibody titer and cytotoxic T-lymphocyte response for co-immunization were increased compared with that of pcDNA3.1-mip immunization. Statistical analysis using one-way analysis of variance (ANOVA) showed that there was a significant difference between the groups (P<0.01). The results indicated that the ctxB gene enhanced the humoral and cellular immune response to the mip gene immunization. These findings provide experimental evidence to support the development of the L. pneumophila DNA vaccine.


Subject(s)
Cholera Toxin/biosynthesis , Cholera Toxin/immunology , Immunophilins/biosynthesis , Immunophilins/immunology , Legionella pneumophila/metabolism , Membrane Proteins/biosynthesis , Membrane Proteins/immunology , Peptidylprolyl Isomerase/biosynthesis , Peptidylprolyl Isomerase/immunology , Protein Engineering/methods , Vibrio cholerae/metabolism , Adjuvants, Immunologic/biosynthesis , Adjuvants, Immunologic/genetics , Animals , Bacterial Proteins , Cholera Toxin/genetics , Female , Immunity, Cellular/immunology , Immunophilins/genetics , Immunophilins/therapeutic use , Legionella pneumophila/genetics , Legionnaires' Disease/prevention & control , Membrane Proteins/genetics , Membrane Proteins/therapeutic use , Mice , Mice, Inbred BALB C , Peptidylprolyl Isomerase/genetics , Peptidylprolyl Isomerase/therapeutic use , Recombinant Proteins/biosynthesis , Recombinant Proteins/immunology , Vibrio cholerae/genetics , Viral Vaccines/biosynthesis , Viral Vaccines/genetics , Viral Vaccines/immunology , Viral Vaccines/therapeutic use
19.
Acta Biochim Biophys Sin (Shanghai) ; 36(9): 629-36, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15346201

ABSTRACT

Basic peptides such as human immunodeficiency virus type 1 (HIV-1) Tat-(48-60) and Drosophila Antennapedia-(43-58) have been reported to have a membrane permeability and a carrier function for intracellular protein delivery. Based on the fluorescence microscopic observations of the vascular endothelial cells (ECV-304) and the primary cultured neuroglial cells, we found that human Clock protein DNA-binding peptide [residue 35-47, hClock-(35-47)] had a translocation activity very similar to Tat-(48-60). The cellular uptake of hClock-(35-47) increases with the increase of incubation time and concentration. The internalization effect at 4 degrees was same as that at 37 degrees C. Internalization of hClock-(35-47) was saturable and could be inhibited by the excess of the other MPPs. Moreover, the uptake of these peptides were significantly inhibited in the presence of heparan sulfate. These results strongly suggested that the hClock-(35-47) shared a common or very similar internalization pathway with other MPPs. Furthermore, we injected rat through the common carotid artery with hClock-(35-47)-FITC peptide, and cryostat sections of the brain were prepared and observed using a fluorescence microscope. Result showed that the peptide had the ability to translocate through the blood-brain barrier. It is promising to provide a new safe carrier for the intracellular and encephalic treatment.


Subject(s)
Brain/metabolism , Cell Membrane Permeability , Cell Membrane/metabolism , DNA-Binding Proteins/metabolism , Peptides/metabolism , Proteins/chemistry , Amino Acid Sequence , Animals , Blood-Brain Barrier , Brain/cytology , Cells, Cultured , Endothelium, Vascular/cytology , HeLa Cells , Heparitin Sulfate/pharmacology , Humans , Kinetics , Microscopy, Fluorescence , Neuroglia/cytology , Neuroglia/metabolism , Peptides/chemical synthesis , Peptides/chemistry , Peptides/drug effects , Rats
20.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 35(2): 198-200, 2004 Mar.
Article in Chinese | MEDLINE | ID: mdl-15071915

ABSTRACT

OBJECTIVE: To investigate the growth-inhibiting and apoptosis-inducing effects of isorhmnetin on HeLa cells and to disclose the role of telomerase activity of tumor cells. METHODS: The methods of cell culture in vitro were adopted. HeLa cells were treated with isorhmnetin in different concentrations for 2 days, and then were observed and analyzed by use of MTT, Flow-Cytometry (FCM) and TRAP-ELIAS technique for inspecting the HeLa cells' growth and telomerase activity. RESULTS: The growth of HeLa cells was inhibited evidently after treatment by isorhmnetin. The rate of apoptosis was 31.7% after the HeLa cells were treated with 20 micrograms/ml isorhmnetin. Isorhmnetin could inhibit the activity of telomerase. CONCLUSION: By inhibiting the activity of telomerase, isorhmnetin can inhibit the growth of HeLa cells.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Flavonols/pharmacology , Telomerase/metabolism , Cell Division/drug effects , HeLa Cells , Humans , Quercetin/analogs & derivatives , Telomerase/drug effects
SELECTION OF CITATIONS
SEARCH DETAIL
...