ABSTRACT
INTRODUCTION: Pulmonary embolism (PE) is a potentially fatal complication and its morbidity together with fatalness will further increase when in patients with malignant tumors. Fast and accurate early diagnosis of PE thus seems considerably important. OBJECTIVE: To explore the risk factors of lung cancer complicated with PE. MATERIALS AND METHODS: A retrospective cohort study consisted of 40 lung cancer patients with PE (PE group) and 60 lung cancer patients without PE (non-PE group) were analyzed. RESULTS: The white blood cell (WBC) count, D-dimer and low-density lipoprotein (LDL) were higher in PE group than those in non-PE group (P < 0.05), whereas the arterial partial pressure of oxygen (PaO2 ) in PE group was lower than that in non-PE group (P < 0.05). Carcinoembryonic antigen (CEA) level between two groups also exhibited statistical difference (P < 0.05). Those lung adenocarcinoma patients with stages III and IV tumor, coupled with deep venous thrombosis (DVT), having experienced bevacizumab treatment or platinum-based chemotherapy more likely suffered from PE (P < 0.05). The multivariate analysis revealed that high D-dimer, chemotherapy, DVT, stages III to IV, adenocarcinoma were independent risk factors associated with PE (P < 0.05). The overall survival time of patients in case group was significantly shorter than that in the control group with a median survival duration being 10.5 months (95%CI, 8.95-12.05) and 16.8 months (95%CI, 14.62-18.98), respectively, (P < 0.01). CONCLUSIONS: High D-dimer, chemotherapy, DVT, stages III to IV and adenocarcinoma might have a positive correlation with PE, meanwhile, PE always predicted a poor prognosis in lung cancer patients.
Subject(s)
Adenocarcinoma of Lung , Lung Neoplasms , Pulmonary Embolism , Humans , Lung Neoplasms/complications , Lung Neoplasms/epidemiology , Pulmonary Embolism/complications , Pulmonary Embolism/diagnosis , Pulmonary Embolism/epidemiology , Retrospective Studies , Risk FactorsABSTRACT
Biohydrometallurgy is generally considered as a green technology for the recycling of industrial solid waste. In this study, an indigenous fungal strain named Y5 with the ability of high-yielding organic acids was isolated and applied in bioleaching of waste printed circuit boards (PCBs). The strain Y5 was identified as Penicillium chrysogenum by morphological and molecular identification. Meanwhile, we investigated that an optimal set of culturing conditions for the fungal growth and acids secretion was 15 g/L glucose with initial pH 5.0, temperature 25°C and shaking speed 120 rpm in shaken flasks culture. Moreover, three bioleaching processes such as one-step, two-step and spent medium processes were conducted to extract copper from waste PCBs. Spent medium bioleaching showed higher copper extraction percentage and it was 47% under 5%(w/v) pulp density. Transmission electron microscope (TEM) observation combining with energy dispersive analysis of X-rays (EDAX) showed that the leached metal ions did not obviously damage the hypha cells. All above results indicated that P.chrysogenum strain Y5 has the tolerance to metal ions, suggesting its potential in recycling of metals from waste PCBs in industry.
Subject(s)
Copper/pharmacokinetics , Electronic Waste , Industrial Waste , Penicillium chrysogenum/isolation & purification , Penicillium chrysogenum/metabolism , Recycling/methods , Biodegradation, Environmental , Copper/analysis , Copper/isolation & purification , Green Chemistry Technology/methods , Metallurgy/methods , Metals, Heavy/chemistry , Metals, Heavy/isolation & purification , Metals, Heavy/pharmacokinetics , Microscopy, Electron, Transmission , Penicillium chrysogenum/cytology , Soil Pollutants/chemistry , Soil Pollutants/isolation & purification , Soil Pollutants/pharmacokineticsABSTRACT
To study the effect of heavy metal pollution on microbial communities, microbial diversity and community structure of soils near Qingshuitang industrial district in Zhuzhou, China, were analyzed by Illumina high-throughput sequencing technology. In this study, the microbial diversity and community relative abundance decreased with increased heavy metal pollution level. Proteobacteria (49.56%) were the most abundant phylum in all samples, followed by Chloroflexi (13.07%) and Acidobacteria (8.77%). The microbial community structures of all samples were similar. The overlap of OTUs was 52.64%, and the structures of the abundant OTUs subcommunities were more similar than the rare OTUs community structures were. Heavy metals caused increases in Proteobacteria and Chloroflexi and decreases in Acidobacteria and Nitrospirae. According to Spearman's correlation, Proteobacteria was significantly negatively correlated with Cr, whereas Chloroflexi was positively correlated with Cr. Cd, Cu, Pb, and Zn were significantly negatively correlated with Nitrospirae. These results showed that heavy metal pollution is an important factor affecting the soil microbial structure in the soil of the Qingshuitang industrial district.
Subject(s)
Bacteria/classification , Metals, Heavy/analysis , Microbiota , Soil Microbiology , Soil Pollutants/analysis , China , IndustryABSTRACT
To seek a feasible technique for processing waste printed circuit boards (PCBs), pretreatment of PCBs by table separation and further bioleached by moderate thermophiles in a stirred tank reactor were investigated. The shaking table separation, conducted after grinding and sieving of PCBs, produced two fractions: metal-rich parts (RPCBs), which is more suitable for pyrometallurgy process than untreated PCBs, and metal-poor parts (PPCBs) with only 8.83% metals was then bioleached by a mixed culture of moderate thermophiles effectively. After adaptation, the mixed culture could tolerate 80 g/L PPCBs. The bioleaching results showed that metals recovery was 85.23% Zn, 76.59% Cu and 70.16% Al in only 7 days. Trace Pb and Sn were detected in the leachate because of precipitating. The microorganism community structure was analyzed by amplified ribosomal DNA restriction analysis. Two moderately thermophilic bacteria species were identified as Leptospirillum ferriphilum and Acidithiobacillus caldus. Furthermore, uncultured Thermoplasmatales archaeon was also detected in the leaching system. It was also shown that moderate thermophiles revealed best bioleaching ability when compared with mesophiles and the mixture of mesophiles and moderate thermophiles. Finally, we designed a two-stage process model according to the present study to achieve semi-industrial waste PCBs recycling and economic feasibility analysis indicated that the process was profitable.
Subject(s)
Acidithiobacillus/metabolism , Bioreactors/microbiology , Electronic Waste , Metals/metabolism , Recycling , Temperature , Metals/isolation & purificationABSTRACT
BACKGROUND: To investigate the value of Ubiquitin specific peptidase 8 (USP8), Chitinase 3-like 1 (YKL40), Heat shock protein 90a (HSP90α), glutathione S-transferase P1 (GSTP1), carcinoembryonic antigen (CEA), neuron specific enolase (NSE) and cytokeratin fragment antiogen 21-1 (CYFRA21-1) in bronchoalveolar lavage fluid (BALF) and serum for diagnosis in patients with peripheral lung cancer. METHODS: The concentration of these markers were measured in 50 patients with peripheral lung cancer and 50 patients with benign lung diseases by using enzyme-linked immuno sorbent assay methods. RESULTS: There were significant differences between the peripheral lung cancer group and the benign lung disease group (P < 0.05) in the BALF of USP8, YKL40, HSP90α, CEA, NSE and CYFRA21-1. There were significant differences between the peripheral lung cancer group and the benign lung disease group (P < 0.05) in the serum of HSP90α and CEA. There were no differences in others. There were no correlation between the concentration of all markers and age, histological type, TNM stage (I-IV). There was a weak correlation between the primary foci diameters and the concentration of YKL40 in BALF. (Pearson's correlation: 0.203, P = 0.048) The diagnostic efficiencies of USP8, YKL40, HSP90α were superior to CYFRA21-1 and NSE, being lower CEA. CONCLUSION: Detection of tumor markers in BALF was superior to serum specimens. The measurement of USP8, HSP90α and YKL40 in BALF had more clinical value for the diagnosis of peripheral pulmonary carcinoma.
Subject(s)
Biomarkers, Tumor/blood , Bronchoalveolar Lavage Fluid/chemistry , Carcinoma, Squamous Cell/pathology , Lung Neoplasms/pathology , Adult , Aged , Antigens, Neoplasm/blood , Antigens, Neoplasm/metabolism , Biomarkers, Tumor/metabolism , Carcinoembryonic Antigen/blood , Carcinoembryonic Antigen/metabolism , Carcinoma, Squamous Cell/blood , Carcinoma, Squamous Cell/metabolism , Chitinase-3-Like Protein 1/metabolism , Endopeptidases/metabolism , Endosomal Sorting Complexes Required for Transport/metabolism , Female , Glutathione S-Transferase pi/blood , Glutathione S-Transferase pi/metabolism , HSP90 Heat-Shock Proteins/blood , HSP90 Heat-Shock Proteins/metabolism , Humans , Keratin-19/blood , Keratin-19/metabolism , Lung Neoplasms/blood , Lung Neoplasms/metabolism , Male , Middle Aged , Neoplasm Staging , Phosphopyruvate Hydratase/blood , Phosphopyruvate Hydratase/metabolism , Ubiquitin Thiolesterase/metabolismABSTRACT
This study assessed the modulating effects of liquiritin against cognitive deficits, oxidative damage, and neuronal apoptosis induced by subsequent bilateral intrahippocampal injections of aggregated amyloid-ß1-42 (Aß1-42). This study also explored the molecular mechanisms underlying the above phenomena. Liquiritin was orally administered to rats with Aß1-42-induced cognitive deficits for 2 weeks. The protective effects of liquiritin on the learning and memory impairment induced by Aß1-42 were examined in vivo by using Morris water maze. The rats were then euthanized for further studies. The antioxidant activities of liquiritin in the hippocampus of the rats were investigated by biochemical and immunohistochemical methods. The apoptosis of the neurons was assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay. Liquiritin at doses of 50-100 mg/kg significantly improved the cognitive ability, restored the abnormal activities of glutathione peroxidase and superoxide dismutase, and decreased the levels of malondialdehydeï¼8-hydroxy-2'-deoxyguanosine and protein carbonyl in the hippocampus of rats with Alzheimer's disease. Moreover, neural apoptosis in the hippocampus of Aß1-42-treated rats was reversed by liquiritin. Liquiritin can significantly ameliorate Aß1-42-induced spatial learning and memory impairment by inhibiting oxidative stress and neural apoptosis.
Subject(s)
Flavanones/pharmacology , Glucosides/pharmacology , Hippocampus/drug effects , Neuroprotective Agents/pharmacology , 8-Hydroxy-2'-Deoxyguanosine , Alzheimer Disease/drug therapy , Amyloid beta-Peptides , Animals , Apoptosis/drug effects , Cognition , Cognition Disorders , Deoxyguanosine/analogs & derivatives , Flavanones/chemistry , Glucosides/chemistry , Glutathione Peroxidase/metabolism , Male , Malondialdehyde/pharmacology , Maze Learning/drug effects , Molecular Structure , Neurons/metabolism , Oxidative Stress/drug effects , Peptide Fragments/pharmacology , Rats , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolismABSTRACT
BACKGROUND AND AIMS: Tuberculosis, a chronic infectious disease caused by Mycobacterium tuberculosis, may invade all organs but mainly affect lungs. Most hepatic tuberculosis could be a part of systemic miliary tuberculosis. METHODS: We reported a case of pulmonary tuberculosis combined with hepatic tuberculosis and reviewed the relevant literature. RESULTS: A 40-year-old Chinese male with fatigue for half a year and cough as well as night sweat for 2 months was admitted to our hospital. The chest computed tomography (CT) showed multiple nodules combined with bronchial stenosis and lymphadenectasis in the mediastina at the right hilum of lung. The epigastrium CT showed lumps in the liver and retroperitoneal lymphadenectasis in the peritoneal cavity. The abdominal color Doppler ultrasound revealed lumps in the liver. The lung and liver puncture biopsy revealed granulomatous lesions, chronic inflammatory changes in the strip-like fibrous tissues and plenty of caseification, all of which suggest the diagnosis of tuberculosis. CONCLUSION: Hepatic tuberculosis is usually associated with atypical clinical manifestations. Imageological examination combined with imaging-guided fine needle aspiration biopsy may be the best method for the confirmed diagnosis.
Subject(s)
Tuberculosis, Hepatic/drug therapy , Tuberculosis, Pulmonary/drug therapy , Adult , Antitubercular Agents/therapeutic use , Comorbidity , Diagnosis, Differential , Humans , Male , Tomography, X-Ray Computed , Treatment Outcome , Tuberculosis, Hepatic/complications , Tuberculosis, Hepatic/diagnostic imaging , Tuberculosis, Pulmonary/complications , Tuberculosis, Pulmonary/diagnostic imaging , Ultrasonography, Doppler, ColorABSTRACT
BACKGROUND: Since the first genome-wide association study report of an association between the ORMDL3 rs7216389 polymorphism and asthma, many studies have been carried out to establish its role in asthma susceptibility among different ethnic groups. However, results have not been consistent across all studies, compelling us to conduct the present meta-analysis. METHODS: A literature search for eligible studies published before January 20, 2014 was conducted in the MEDLINE, EMBASE, and CNKI databases. The association was assessed using pooled crude odds ratios (ORs) with their corresponding 95% confidence intervals (CIs). RESULTS: A total of 18 individual studies in 15 publications (total 7904 asthma patients and 10,874 healthy controls) were included in the meta-analysis. A meta-analysis of all included studies suggested that there was a highly significant risk effect conferred by the rs7216389*T allele on asthma susceptibility. In addition, we performed stratified analyses to evaluate ethnicity-specific and age-specific effects. Our subgroup analyses based on ethnicity and age-of-onset confirmed the role of the ORMDL3 rs7216389 polymorphism in conferring susceptibility to both childhood- and adult-onset asthma, especially in Caucasians and Asians. CONCLUSIONS: The results of this meta-analysis firmly established that genetic variation at the rs7216389 locus, which controls the expression of the ORMDL3, may be a major, independent predisposing factor for asthma in ethnically diverse populations. However, further systematic studies are needed to determine the underlying mechanisms of this association.
Subject(s)
Asthma/genetics , Genetic Predisposition to Disease/genetics , Genetic Variation , Membrane Proteins/genetics , Age of Onset , Alleles , Asian People/genetics , Asthma/epidemiology , Asthma/ethnology , Gene Frequency , Genetic Predisposition to Disease/ethnology , Genotype , Humans , Odds Ratio , Polymorphism, Single Nucleotide , Risk Factors , White People/geneticsABSTRACT
Studies have suggested that bone marrow-derived mesenchymal stem cells (MSCs) may be used as a tool for gene therapy. Developmental endothelial locus-1 (Del-1) is a critical factor for cell migration and infiltration via the inhibition of the function of a major leukocyte adhesion receptor LFA-1 which prevents leukocyte adhesion to the endothelium. In the present study, we hypothesized that MSC-based Del-1 gene therapy may have potential therapeutic applications for lipopolysaccharide (LPS)-induced lung injury. The MSCs in the present assay were isolated from 6 week-old male mice. In order to investigate the therapeutic effect of the Del-1 gene on LPS-induced ALI mice, a lentivirus vector containing the Del-1 gene was constructed and transduced into the MSCs. In the in vivo assay, we induced lung injury with LPS injection and treated mice with different groups of MSCs, and compared with groups treated with MSCs alone, we observed that the administration with MSCs carrying Del-1 (MSCs-Del1) markedly alleviated the LPS-induced lung injury. There were significant decreases in the number of neutrophils in bronchoalveolar lavage (BAL) and the serum levels of TNF-α and IL-6 in the Del-1-expressed MSC-treated mice. Furthermore, compared with MSCs treated alone, Del1-MSC-treated mice also exhibited low lung injury scores, high protein concentrations and myeloperoxidase activity. In conclusion, treatment with Del-1-expressed MSCs significantly decreases the severity of endotoxin-induced acute lung injury and the level of inflammatory cytokines in mice.
Subject(s)
Acute Lung Injury/genetics , Acute Lung Injury/therapy , Carrier Proteins/genetics , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/metabolism , Acute Lung Injury/chemically induced , Animals , Bronchoalveolar Lavage Fluid/chemistry , Calcium-Binding Proteins , Carrier Proteins/metabolism , Cell Adhesion Molecules , Female , Gene Expression , Genetic Therapy , Genetic Vectors/genetics , Intercellular Signaling Peptides and Proteins , Interleukin-6/metabolism , Lipopolysaccharides/adverse effects , Lung/metabolism , Lung/pathology , Male , Mice , Peroxidase/metabolism , Retroviridae/genetics , Transduction, Genetic , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND: Angptl4 is a secreted protein involved in the regulation of vascular permeability, angiogenesis, and inflammatory responses in different kinds of tissues. Increases of vascular permeability and abnormality changes in angiogenesis contribute to the pathogenesis of tumor metastasis, ischemic-reperfusion injury. Inflammatory response associated with Angptl4 also leads to minimal change glomerulonephritis, wound healing. However, the role of Angptl4 in vascular permeability, angiogenesis, and inflammation is controversy. Hence, an underlying mechanism of Angptl4 in different kind of tissues needs to be further clarified. METHODS: Keywords such as angptl4, vascular permeability, angiogenesis, inflammation, and endothelial cells were used in search tool of PUBMED, and then the literatures associated with Angptl4 were founded and read. RESULTS: Data have established Angptl4 as the key modulator of both vascular permeability and angiogenesis; furthermore, it may also be related to the progression of metastatic tumors, cardiovascular events, and inflammatory diseases. This view focuses on the recent advances in our understanding of the role of Angptl4 in vascular permeability, angiogenesis, inflammatory signaling and the link between Angptl4 and multiple diseases such as cancer, cardiovascular diseases, diabetic retinopathy, and kidney diseases. CONCLUSIONS: Taken together, Angptl4 modulates vascular permeability, angiogenesis, inflammatory signaling, and associated diseases. The use of Angptl4-modulating agents such as certain drugs, food constituents (such as fatty acids), nuclear factor (such as PPARα), and bacteria may treat associated diseases such as tumor metastasis, ischemic-reperfusion injury, inflammation, and chronic low-grade inflammation. However, the diverse physiological functions of Angptl4 in different tissues can lead to potentially deleterious side effects when used as a therapeutic target. In this regard, a better understanding of the underlying mechanisms for Angptl4 in different tissues is necessary.
Subject(s)
Angiopoietins/metabolism , Capillary Permeability/physiology , Inflammation/metabolism , Angiopoietin-Like Protein 4 , Animals , Humans , Neoplasms/metabolism , Neoplasms/pathology , Neovascularization, Pathologic/metabolism , Reperfusion Injury/metabolismABSTRACT
The aim of our study was to investigate the chronic obstructive pulmonary disease (COPD) assessment test (CAT), serum copeptin, procalcitonin and C-reactive protein (CRP) levels as potential predictive factors for recurrence of acute exacerbation and all-cause mortality in 6 months of COPD inpatients. One hundred fifty-nine patients who met the inclusion criteria were enrolled and followed up for 6 months. The CAT scores, serum copeptin, procalcitonin and CRP levels were measured on admission and 14 days and 3 months later in all patients. The primary endpoint was recurrence of acute exacerbation in 6 months. The secondary endpoint was all-cause mortality after 6 months. The CAT scores, serum copeptin, procalcitonin and CRP levels were significantly elevated on admission and stabilized at 14 days (P < 0.01). In a univariate logistic regression analysis, CAT scores (odds ratio [OR] = 1.10), forced expiratory volume in 1 second % (OR = 1.01), serum copeptin (OR = 1.32) and CRP levels (OR = 1.01) were significantly related to recurrence of acute exacerbation in 6 months (P < 0.05). In a multivariate logistic regression model, increasing CAT scores (OR = 1.10) and serum copeptin levels (OR = 1.29) were still associated with an increased odds of exacerbation (P < 0.05). In a univariate logistic regression analysis, increasing CAT scores (OR = 1.19), forced expiratory volume in 1 second % (OR = 1.05), serum copeptin levels (OR = 1.44) and hospitalization in the previous years (OR = 1.24) were significant determinants of death over a follow-up period of 6 months (P < 0.05). But only serum copeptin (OR = 1.53) and CAT scores (OR = 1.37) were associated with mortality in multivariate logistic regression analysis. Hence, high CAT scores and serum copeptin levels link with recurrence of acute exacerbation and all-cause mortality during 6 months in patients with acute exacerbation of COPD.
Subject(s)
Glycopeptides/blood , Pulmonary Disease, Chronic Obstructive/blood , Pulmonary Disease, Chronic Obstructive/diagnosis , Respiratory Function Tests/standards , Acute Disease , Aged , Aged, 80 and over , Biomarkers/blood , Female , Follow-Up Studies , Humans , Male , Middle Aged , Protein Precursors/blood , Pulmonary Disease, Chronic Obstructive/mortality , Recurrence , Respiratory Function Tests/methods , Surveys and QuestionnairesABSTRACT
Rhodococcus sp. JDC-11, capable of utilizing di-n-butyl phthalate (DBP) as the sole source of carbon and energy, was isolated from sewage sludge and confirmed mainly based on 16S rRNA gene sequence analysis. The optimum pH, temperature, and agitation rate for DBP degradation by Rhodococcus sp. JDC-11 was 8.0, 30 degrees C, and 175 rpm, respectively. In addition, the effect of glucose concentration on DBP degradation indicated that low concentration of glucose inhibited the degradation of DBP while high concentrations of glucose increased its degradation. Meanwhile, the substrates utilization test showed that JDC-11 could also utilize other phthalates. Furthermore, the major metabolites of DBP degradation were identified as mono-butyl phthalate and phthalic acid by gas chromatography-mass spectrometry and the metabolic pathway of DBP degradation by Rhodococcus sp. JDC-11 was tentatively speculated. Using a set of new degenerate primer, partial sequence of the 3, 4-phthalate dioxygenase gene was obtained from the strain. Sequence analysis revealed that the phthalate dioxygenase gene of JDC-11 was highly homologous to the large subunit of phthalate dioxygenase from Rhodococcus coprophilus strain G9.
Subject(s)
Bacterial Proteins/genetics , Dibutyl Phthalate/metabolism , Oxygenases/genetics , Rhodococcus/metabolism , Bacterial Proteins/metabolism , Biodegradation, Environmental , Molecular Sequence Data , Oxygenases/metabolism , Phylogeny , Rhodococcus/classification , Rhodococcus/genetics , Rhodococcus/isolation & purification , SewageABSTRACT
The serum samples and corresponding cervical swabs were collected from 50 women with genital warts from Tianjin city, China. The neutralizing antibodies against HPV-16, -18, -58, -45, -6 and -11 in serum samples were tested by using pseudovirus-based neutralization assays and HPV DNAs in cervical swabs were also tested by using a typing kit that can detect 21 types of HPV. The results revealed that 36% (18/50) of sera were positive for type-specific neutralizing antibodies with a titer range of 160-2560, of which 22%(11/50), 12%(6/50), 10%(5/50), 4%(2/50), 4%(2/50) and 2%(1/50) were against HPVs -6, -16, -18, -58, -45 and -11, respectively. Additionally, 60% (30/50) of samples were HPV DNA-positive, in which the most common types detected were HPV-68(18%), HPV-16(14%), HPV-58(12%), HPV-33(8%) and HPV-6, HPV-11, HPV-18 and HPV-52 (6% each). The concordance between HPV DNA and corresponding neutralizing antibodies was 56% (28/50) with a significant difference (P<0.05). The full-length sequences of five HPV types (HPV -42, -52, -53, -58 and -68) were determined and exhibited 98%-100% identities with their reported genomes. The present data may have utility for investigating the natural history of HPV infection and promote the development of HPV vaccines.
Subject(s)
Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Condylomata Acuminata/virology , Papillomaviridae/classification , Papillomaviridae/isolation & purification , Blood/immunology , Cervix Uteri/virology , China , Cluster Analysis , DNA, Viral/chemistry , DNA, Viral/genetics , Female , Genotype , Humans , Molecular Epidemiology , Molecular Sequence Data , Neutralization Tests/methods , Papillomaviridae/genetics , Papillomaviridae/immunology , Phylogeny , Prevalence , Sequence Analysis, DNA , Virology/methodsABSTRACT
A gram negative isolate designated JDC-41 was obtained from river sludge using mixtures of phthalate esters as the sole source and energy. The isolate was identified as Ochrobactrum sp. based on its 16S rRNA gene sequence. Over 87% of supplied di-n-butyl phthalate (DBP) was degraded by JDC-41 in a pH neutral mineral salts medium at 30 degrees C within 48 h. Increased DBP (50-500 mg/L) in the culture correspondingly increased degradation half-life from 3.83 to 18.12 h. DBP induced cells more rapidly degraded DBP.
Subject(s)
Dibutyl Phthalate/metabolism , Ochrobactrum/metabolism , Plasticizers/metabolism , Water Pollutants, Chemical/metabolism , Base Sequence , Molecular Sequence Data , Ochrobactrum/genetics , Ochrobactrum/isolation & purificationABSTRACT
Four di-butyl-phthalate(DBP)-degrading bacterial strains, JDC-1, JDC-8, JDC-9 and JDC-12, were isolated from soil. The strains were gram positive. The 16S rRNA sequence analysis revealed that the four strains had similarities of 99% with Arthrobacter sp.. According to the morphologic, physiobiochemical characteristics and the analysis of their 16S rRNA, all the four strains were identified as Arthrobacter sp.. A 900 bp DNA fragment was obtained from the four strains by PCR amplified and clone. When compared with the large subunit of phthalate dioxygenase gene (phtA) of Arthrobacter keyseri, more than 96% similarities were evident in the nucleotide sequences. The optimal growth conditions and degradation rates of DBP were tested and the result indicated that the optimal growth conditions of the four bacteria strains were pH 7.0-8.5 and 30-35 degrees C. All the four bacteria strains performed efficiently for DBP degrading capabilities under optimal conditions. The most efficient strain JDC-1 degraded 500 mg/L DBP completely within 28 h whereas the least efficient strain JDC-8 degraded 500 mg/L DBP completely within 40 h. This study is helpful to the investigation of DBP-degrading mechanisms and the development of microbial resources.
Subject(s)
Arthrobacter/genetics , Bacterial Proteins/genetics , Dibutyl Phthalate/metabolism , Oxygenases/genetics , Soil Pollutants/metabolism , Amino Acid Sequence , Arthrobacter/enzymology , Arthrobacter/growth & development , Arthrobacter/isolation & purification , Bacterial Proteins/metabolism , Biodegradation, Environmental , Cloning, Molecular , Genes, Bacterial , Molecular Sequence Data , Oxygenases/metabolism , RNA, Ribosomal, 16S/genetics , Soil MicrobiologyABSTRACT
To find out whether the mutations of HIV-1 Env have influence on the assembly of pseudovirus and their abilities to infect cells, site-directed mutation (A457D)was performed using cycling mutagenesis and selection of mutants with DpnI. Transformation and plasmid purification technologies were used to obtain mutated env clone. Then both the prototype and the mutant were co-transfected with pSG3(delta(env)) to 293FT cells, respectively. Single-cycle infection assay was employed to analyze the effect of the prototype and the mutant on the ability of functional pseudovirus assembly. The transient expression of both the prototype S12-42-1 and mutant S12-42M were confirmed by Western blot essay. The S/CO value was less than 1 for S12-42-1 and 6.65 for S12-42M, demonstrating the functional pseudovirus was generated only for S12-42M. So mutation on HIV-1 Env has influence on the assembly of pseudovirus and their abilities to infect cells.
Subject(s)
HIV Infections/virology , HIV-1/physiology , Mutation , Virion/physiology , Virus Assembly , env Gene Products, Human Immunodeficiency Virus/genetics , Amino Acid Sequence , Base Sequence , Cell Line , HIV-1/chemistry , HIV-1/genetics , Humans , Molecular Sequence Data , Sequence Alignment , Virion/genetics , env Gene Products, Human Immunodeficiency Virus/chemistry , env Gene Products, Human Immunodeficiency Virus/metabolismABSTRACT
In order to evaluate the immunogenicity of HPV 58 L1 DNA vaccines, five DNA vaccines had been constructed with pcDNA3.1 vector containing different L1 genes of HPV 58, which were designated as L1h, L1hDeltac, L1S, L1SM and L1wt. The protein expression of DNA vaccines in vitro was tested by Western blot. The ability of forming pseudovirus was evaluated by transfecting DNA vaccine together with pcDNA3.1-h58L2 and pcDNA3.1-GFP into 293FT cells. The neutralizing antibodies and cellular immune response produced in BALB/c mice immunized with the DNA vaccines were detected by using pseudovirus-based neutralization assay and ELISPOT respectively. The results showed that the five DNA vaccines had been successfully constructed; the level of protein expression of L1hDeltac was the highest and those for L1S and L1SM were of medium, while no expressed target protein of L1wt was detected. Only L1S could form the pseudovirus while the other four vaccines could not. L1S and L1h could induce neutralizing antibody. However, the average titer of neutralizing antibody for L1S (1:6,400) was much higher than that for L1h (1:48) and the other three vaccines could not induce neutralizing antibody. No cellular immune response for all five DNA vaccines was detectable by ELISPOT. The results indicated that DNA vaccine against HPV 58 can form pseudovirus in vitro, also can induce high level of neutralizing antibodies. This provides reference for screening HPV vaccine in future.