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1.
J Hazard Mater ; 471: 134437, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38691934

ABSTRACT

Crude oil is a hazardous pollutant that poses significant and lasting harm to human health and ecosystems. In this study, Moesziomyces aphidis XM01, a biosurfactant mannosylerythritol lipids (MELs)-producing yeast, was utilized for crude oil degradation. Unlike most microorganisms relying on cytochrome P450, XM01 employed two extracellular unspecific peroxygenases, MaUPO.1 and MaUPO.2, with preference for polycyclic aromatic hydrocarbons (PAHs) and n-alkanes respectively, thus facilitating efficient crude oil degradation. The MELs produced by XM01 exhibited a significant emulsification activity of 65.9% for crude oil and were consequently supplemented in an "exogenous MELs addition" strategy to boost crude oil degradation, resulting in an optimal degradation ratio of 72.3%. Furthermore, a new and simple "pre-MELs production" strategy was implemented, achieving a maximum degradation ratio of 95.9%. During this process, the synergistic up-regulation of MaUPO.1, MaUPO.1 and the key MELs synthesis genes contributed to the efficient degradation of crude oil. Additionally, the phylogenetic and geographic distribution analysis of MaUPO.1 and MaUPO.1 revealed their wide occurrence among fungi in Basidiomycota and Ascomycota, with high transcription levels across global ocean, highlighting their important role in biodegradation of crude oil. In conclusion, M. aphidis XM01 emerges as a novel yeast for efficient and eco-friendly crude oil degradation.


Subject(s)
Biodegradation, Environmental , Glycolipids , Mixed Function Oxygenases , Petroleum , Surface-Active Agents , Petroleum/metabolism , Surface-Active Agents/metabolism , Surface-Active Agents/chemistry , Glycolipids/metabolism , Mixed Function Oxygenases/metabolism , Mixed Function Oxygenases/genetics , Polycyclic Aromatic Hydrocarbons/metabolism , Polycyclic Aromatic Hydrocarbons/chemistry , Alkanes/metabolism
2.
Nat Commun ; 15(1): 1995, 2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38443404

ABSTRACT

Cardiac macrophage contributes to the development of cardiac fibrosis, but factors that regulate cardiac macrophages transition and activation during this process remains elusive. Here we show, by single-cell transcriptomics, lineage tracing and parabiosis, that cardiac macrophages from circulating monocytes preferentially commit to macrophage-to-myofibroblast transition (MMT) under angiotensin II (Ang II)-induced hypertension, with accompanying increased expression of the RNA N6-methyladenosine demethylases, ALKBH5. Meanwhile, macrophage-specific knockout of ALKBH5 inhibits Ang II-induced MMT, and subsequently ameliorates cardiac fibrosis and dysfunction. Mechanistically, RNA immunoprecipitation sequencing identifies interlukin-11 (IL-11) mRNA as a target for ALKBH5-mediated m6A demethylation, leading to increased IL-11 mRNA stability and protein levels. By contrast, overexpression of IL11 in circulating macrophages reverses the phenotype in ALKBH5-deficient mice and macrophage. Lastly, targeted delivery of ALKBH5 or IL-11 receptor α (IL11RA1) siRNA to monocytes/macrophages attenuates MMT and cardiac fibrosis under hypertensive stress. Our results thus suggest that the ALKBH5/IL-11/IL11RA1/MMT axis alters cardiac macrophage and contributes to hypertensive cardiac fibrosis and dysfunction in mice, and thereby identify potential targets for cardiac fibrosis therapy in patients.


Subject(s)
Adenine , Hypertension , Interleukin-11 , Animals , Humans , Mice , Adenine/analogs & derivatives , AlkB Homolog 5, RNA Demethylase , Angiotensin II , Cardiotonic Agents , Macrophages , Myofibroblasts , RNA
3.
Front Endocrinol (Lausanne) ; 14: 1216413, 2023.
Article in English | MEDLINE | ID: mdl-37937052

ABSTRACT

Background: Waist circumference can be used as an anthropometric measure to assess central obesity and is easier and more convenient than the waist-to-hip ratio in identifying the risk of obesity and medical problems. Most studies showing an association between obesity and infertility in women have used BMI to measure obesity. Our goal was to examine any potential association between waist circumference and infertility. Methods: This cross-sectional study, which formed part of the National Health and Nutrition Examination Survey (NHANES), comprised women ages 18 to 45 between 2017 and 2020. Participants without waist circumference data or information on infertility were removed from the study. The independent relationship between waist circumference and infertility was investigated using weighted binary logistic regression and subgroup analysis. Results: We investigated 1509 participants and discovered that the prevalence of infertility rose as the WC trisection rose. (tertile 1, 7.55%; tertile 2, 10.56%; tertile 3, 15.28%; trend < 0.001). Multivariate logistic regression showed that after total adjustment, higher WC levels were associated with an increased likelihood of infertility in women (OR1.02; 95% CI 1.01-1.03), and There was a 2% rise in the incidence of infertility for every unit (cm) increased WC. Subgroup analysis and interaction tests showed no significant dependence of the effects of marital status, diabetes, hypertension, and high cholesterol on the association between WC and infertility (p for all interaction tests > 0.05). The inflection point of the positive non-linear relationship between WC and infertility was 116.6 cm. Conclusion: Excessive waist circumference assessment may increase the probability of infertility, and more attention should be paid to the management of waist circumference should be given more attention.


Subject(s)
Infertility, Female , Humans , Female , Waist Circumference , Risk Factors , Nutrition Surveys , Infertility, Female/etiology , Infertility, Female/complications , Cross-Sectional Studies , Body Mass Index , Obesity/complications , Obesity/epidemiology
4.
Huan Jing Ke Xue ; 43(3): 1307-1314, 2022 Mar 08.
Article in Chinese | MEDLINE | ID: mdl-35258194

ABSTRACT

Polycyclic aromatic hydrocarbons (PAHs) play a key role in the formation of secondary organic areole and ozone. This study sampled three commercial Chinese restaurants and a food plant in Shenzhen to analyze the emission characteristics of PAHs, especially the alkyl PAHs in both gas and particle phases. The results showed that the ρ(total PAHs)in the particle and gas phase were (1381.6±140.5) ng·m-3, (1030.2±116.4) ng·m-3, (908.3±111.9) ng·m-3, and (838.0±93.5) ng·m-3 in the food plant, Sichuan, Cantonese, and Zhejiang restaurants, respectively. More than 60% of the PAHs were distributed in the gas phase, especially the lower molecular weight PAHs (lower than Chrysene). The gas phase proportion of naphthalene was the highest, with over 75% of it distributed in the gas phase. However, the PAHs with a higher molecular weight than that of benzo(b)fluorescence were mainly distributed in the particle phase. The total concentration of alkyl PAHs emitted from cooking was much lower than that of the corresponding parent PAHs, and the distribution characteristics of alkyl PAHs were quite different from those of other emission sources. The linear fitting of lgKp and lgPL showed that the slopes of the three commercial restaurants ranged from -0.25 to -0.28, whereas for the food plant, the value was -0.18, which indicates that the gas-particle partitioning of PAHs were not in equilibrium.


Subject(s)
Air Pollutants , Polycyclic Aromatic Hydrocarbons , Air Pollutants/analysis , Cooking , Environmental Monitoring , Polycyclic Aromatic Hydrocarbons/analysis
5.
Reprod Biomed Online ; 43(4): 598-606, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34474977

ABSTRACT

RESEARCH QUESTION: lncRNA IGF2-AS may be related to early pregnancy loss. Does lncRNA IGF2-AS affect trophoblast cell growth? The aim of the present study was to verify that lncRNA IGF2-AS encodes a polypeptide, IGF2-AS-168aa, and to study its role in the pathogenesis of trophoblasts. DESIGN: A small interfering RNA targeted to the IGF2-AS gene (si-IGF2-AS) was designed and transfected into JEG-3 and JAR cells for in-vitro gene silencing. Quantitative polymerase chain reaction and western blotting were used to determine lncRNA IGF2-AS levels in experimental cells. After IGF2-AS suppression, MTT assay was used to assess cell proliferation and apoptosis was determined by flow cytometry. Target gRNA IGF2-AS-gRNA was designed for knockout conducted the corresponding mRNA. HEK293T cells were transfected with the identified positive clone vectors. Finally, IGF2-AS-168aa was analysed by western blotting after the protein-coding region of the IGF2-AS gene was knocked out by CRISPR/Cas9 gene-editing technology. RESULTS: lncRNA IGF2-AS and IGF2-AS-168aa were significantly downregulated in JEG-3 and JAR cells transfected with si-IGF2-AS (lncRNA IGF2-AS: JAR: NC versus small interfering RNA (siRNA)-1: P = 0.019 NC versus siRNA-2: P = 0.013; JEG-3: NC versus siRNA-1: P = 0.001 NC versus siRNA-2: P = 0.004) (IGF2-AS-168aa: JAR: NC versus siRNA-1: P = 0.030 NC versus siRNA-2: P = 0.018; JEG-3: NC versus siRNA-1: P = 0.004 NC versus siRNA-2: P = 0.001). IGF2-AS gene was incapable of encoding IGF2-AS-168aa after the coding region was successfully knocked out in HEK293T cells. Flow cytometry and the MTT assay revealed that IGF2-AS gene silencing led to cell cycle block in the G1 phase, markedly decreasing cell proliferation and increasing apoptosis. CONCLUSION: The IGF2-AS gene encoded a peptide with a potential function in trophoblast cell cycle arrest.


Subject(s)
Abortion, Spontaneous/etiology , Cell Cycle Checkpoints , Proteins/metabolism , Trophoblasts/physiology , Base Sequence , Down-Regulation , Gene Targeting , HEK293 Cells , Humans
6.
Huan Jing Ke Xue ; 42(2): 1013-1022, 2021 Feb 08.
Article in Chinese | MEDLINE | ID: mdl-33742898

ABSTRACT

Specific organic compounds within atmospheric particulate matter are indicators of specific pollution sources and, as such, can be used to differentiate inputs from various air pollution emissions sources in urban areas. Therefore, many studies have been conducted to detect organic particulate matter and screen the associated organic tracers that provide provenance information. This review provides a brief summary of the emission characteristics of biomass burning, cooking, fossil fuel combustion, and traffic. The particular marker compounds that carry provenance information for these four emission sources are discussed and diagnostic ratios are calculated to discuss the use of organic tracers in source apportionment. The shortcomings and new directions of using source tracer screening are also discussed.

7.
Huan Jing Ke Xue ; 41(8): 3467-3474, 2020 Aug 08.
Article in Chinese | MEDLINE | ID: mdl-33124318

ABSTRACT

Cooking is an important source of atmospheric particulate organic matter (POM). In this study, four Chinese restaurants in Shenzhen (west style, dim-sim restaurant, worker's canteen, and Korean cuisine) were sampled to examine the chemical composition of POM and research molecular tracers. The result showed that more than 60% of the PM2.5 mass was due to organic compounds. For the quantified organic compounds, the results indicated that fatty acids, dicarboxylic acids, and n-alkanes were the major organic compounds emitted from all cooking styles, PAHs, sterols, and monosaccharide anhydrides were found at relatively low levels. The composition of POM was strongly influenced by cooking style. The cooking styles of the west and Korean restaurant emitted the most abundant fatty acids, n-alkanes, and PAHs, but the least sterols and monosaccharide anhydrides, whereas the dim-sim restaurant and worker's canteen displayed the opposite results. The values of Fla/(Fla+Pyr) and LG/(Gal+Man) provided candidate tracers for cooking because they were less influenced by the cooking styles and were significantly different from other pollutant sources. Furthermore, cooking contributed significant amounts of fatty acids and dicarboxylic acids to atmospheric PM in Shenzhen.


Subject(s)
Air Pollutants , Polycyclic Aromatic Hydrocarbons , Air Pollutants/analysis , Cooking , Environmental Monitoring , Particulate Matter/analysis , Polycyclic Aromatic Hydrocarbons/analysis
8.
BioData Min ; 13: 10, 2020.
Article in English | MEDLINE | ID: mdl-32863885

ABSTRACT

BACKGROUND: Identifying molecular subtypes of ovarian cancer is important. Compared to identify subtypes using single omics data, the multi-omics data analysis can utilize more information. Autoencoder has been widely used to construct lower dimensional representation for multi-omics feature integration. However, learning in the deep architectures in Autoencoder is difficult for achieving satisfied generalization performance. To solve this problem, we proposed a novel deep learning-based framework to robustly identify ovarian cancer subtypes by using denoising Autoencoder. RESULTS: In proposed method, the composite features of multi-omics data in the Cancer Genome Atlas were produced by denoising Autoencoder, and then the generated low-dimensional features were input into k-means for clustering. At last based on the clustering results, we built the light-weighted classification model with L1-penalized logistic regression method. Furthermore, we applied the differential expression analysis and WGCNA analysis to select target genes related to molecular subtypes. We identified 34 biomarkers and 19 KEGG pathways associated with ovarian cancer. CONCLUSIONS: The independent test results in three GEO datasets proved the robustness of our model. The literature reviewing show 19 (56%) biomarkers and 8(42.1%) KEGG pathways identified based on the classification subtypes have been proved to be associated with ovarian cancer. The outcomes indicate that our proposed method is feasible and can provide reliable results.

9.
Taiwan J Obstet Gynecol ; 59(1): 109-114, 2020 Jan.
Article in English | MEDLINE | ID: mdl-32039776

ABSTRACT

OBJECTIVE: The present study aimed to evaluate insulin-like growth factor 2 antisense (IGF2-AS) in the villi of human embryos and compared its expression between normal pregnancy and early pregnancy loss (EPL). MATERIALS AND METHODS: The present study conducted a microarray analysis to identify the expression profiles of lncRNAs in villi from EPL and normal controls (controls, n = 10 and EPL patients, n = 10). Embryonic villi were collected from women who underwent artificial abortion. QPCR was used to confirm the results. The DNA methylation patterns were analyzed using pyrosequencing and bisulfite sequencing polymerase chain reaction. The percentage of methylation was compared in chorionic villi from the two groups. RESULTS: A total of 57 deregulated differentially expressed lncRNAs were detected, of which 33 were upregulated, and 24 were downregulated. The expression of lncRNA IGF2-AS was downregulated significantly in EPL villi compared with the normal villi. Negative regulation of IGF2-AS may be involved in the development of EPL. Methprimer predicted that IGF2-AS promoter had CpG islands and dense CG sites. Increased methylation at CpG islands present in IGF2-AS gene promoter was observed in EPL villi. CONCLUSION: An increase in methylation of IGF2-AS likely leads to its downregulation in chorionic villi of EPL. The findings suggest that a deficiency of IGF2-AS in the villi is associated with human EPL.


Subject(s)
Abortion, Spontaneous/genetics , Chorionic Villi/metabolism , DNA Methylation/genetics , Proteins/metabolism , RNA, Long Noncoding/metabolism , Adult , Case-Control Studies , CpG Islands/genetics , Down-Regulation/genetics , Female , Humans , Pregnancy , Promoter Regions, Genetic , Up-Regulation/genetics
10.
Curr Med Sci ; 38(3): 505-512, 2018 Jun.
Article in English | MEDLINE | ID: mdl-30074219

ABSTRACT

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy. The ten-eleven translocation (TET) family and 5-hydroxymethylcytosine (5-hmC) were found to be linked with epigenetic reprogramming. The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss (EPL). Embryonic villi were collected from normal pregnant women (control) experiencing medical abortion and from EPL patients at gestation ages of 6, 7 and 8 weeks. The mRNAs of TET family were analysed using quantitative polymerase chain reaction (qPCR), and TET proteins using Western blotting and immunohistochemical analysis. The MethylFlash™ Kit was used to quantify the absolute amount of 5-methylcytosine (5-mC) and 5-hmC. Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age. Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples, which was consistent with the qPCR and Western blot results. The expression of TET1, TET2, and TET3 was lower in the villi in EPL group than in normal pregnancy group (P<0.05 for all). It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo. The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.


Subject(s)
5-Methylcytosine/analogs & derivatives , Abortion, Spontaneous/metabolism , Proto-Oncogene Proteins/metabolism , Trophoblasts/metabolism , 5-Methylcytosine/metabolism , Adult , DNA/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Dioxygenases/genetics , Dioxygenases/metabolism , Female , Humans , Mixed Function Oxygenases/genetics , Mixed Function Oxygenases/metabolism , Pregnancy , Proto-Oncogene Proteins/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Trophoblasts/pathology
11.
Acta Pharmacol Sin ; 38(6): 885-896, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28479604

ABSTRACT

Multidrug resistance (MDR) is a major hurdle in cancer chemotherapy and makes the treatment benefits unsustainable. Combination therapy is a commonly used method for overcoming MDR. In this study we investigated the anti-MDR effect of dihydroartemisinin (DHA), a derivative of artemisinin, in combination with doxorubicin (Dox) in drug-resistant human colon tumor HCT8/ADR cells. We developed a tumor-targeting codelivery system, in which the two drugs were co-encapsulated into the mannosylated liposomes (Man-liposomes). The Man-liposomes had a mean diameter of 158.8 nm and zeta potential of -15.8 mV. In the HCT8/ADR cells that overexpress the mannose receptors, the Man-liposomes altered the intracellular distribution of Dox, resulting in a high accumulation of Dox in the nuclei and thus displaying the highest cytotoxicity (IC50=0.073 µg/mL) among all the groups. In a subcutaneous HCT8/ADR tumor xenograft model, administration of the Man-liposomes resulted in a tumor inhibition rate of 88.59%, compared to that of 47.46% or 70.54%, respectively, for the treatment with free Dox or free Dox+DHA. The mechanisms underlying the anti-MDR effect of the Man-liposomes involved preferential nuclear accumulation of the therapeutic agents, enhanced cancer cell apoptosis, downregulation of Bcl-xl, and the induction of autophagy.


Subject(s)
Antineoplastic Agents/pharmacology , Artemisinins/pharmacology , Colonic Neoplasms/drug therapy , Doxorubicin/pharmacology , Drug Delivery Systems , Drug Resistance, Neoplasm/drug effects , Animals , Antineoplastic Agents/chemistry , Apoptosis/drug effects , Artemisinins/administration & dosage , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Colonic Neoplasms/pathology , Dose-Response Relationship, Drug , Doxorubicin/administration & dosage , Drug Carriers/chemistry , Drug Screening Assays, Antitumor , Female , Humans , Liposomes/chemistry , Mice , Mice, Inbred BALB C , Mice, Nude , Structure-Activity Relationship
12.
Anal Biochem ; 539: 29-32, 2017 12 15.
Article in English | MEDLINE | ID: mdl-28279647

ABSTRACT

Traditional ELISA methods of using animal immunity yield antibodies for detection Cry toxin. Not only is this incredibly harmful to the animals, but is also time-intensive. Here we developed a simple method to yield the recognition element. Using a critical selection strategy and immunoassay we confirmed a clone from the Ph.D-C7C phage library, which has displayed the most interesting Cry1Ab-binding characteristics examined in this study (Fig. 1). The current study indicates that isolating peptide is an alternative method for the preparation of a recognition element, and that the developed assay is a potentially useful tool for detecting Cry1Ab.


Subject(s)
Bacterial Proteins/analysis , Bacteriophages/metabolism , Endotoxins/analysis , Enzyme-Linked Immunosorbent Assay/methods , Hemolysin Proteins/analysis , Peptides/metabolism , Antibodies, Monoclonal/immunology , Bacillus thuringiensis Toxins , Bacterial Proteins/immunology , Endotoxins/immunology , Hemolysin Proteins/immunology , Limit of Detection , Peptide Library , Peptides/isolation & purification
13.
Biomed Pharmacother ; 77: 92-7, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26796271

ABSTRACT

Ovarian cancer (OC) remains one of the most common types of malignant cancer, and the molecular mechanism underlying its proliferation is still largely unclear. It is reported that microRNAs acted as important regulators of cell proliferation by regulating its targeted gene. In this study, our result showed that miR-572 was markedly upregulated in OC cell lines and clinical tissues. Results of both gain-of-function and loss-of-function experiments revealed that upregulation of miR-572 expression dramatically promoted OC cell proliferation, whereas decreased miR-572 expression significantly reduced cell proliferation. Bioinformatics analysis and luciferase reporter assays further revealed PPP2R2C, a putative tumor suppressor as a potential target of miR-572. Moreover, silencing of PPP2R2C using small interfering RNA (siRNA) counteracted the proliferation arrest by miR-572-in in OC cells. In sum, our data provide that miR-572 promoted cell proliferation in OC by targeting PPP2R2C and might serve as a therapeutic target of OC.


Subject(s)
MicroRNAs/genetics , Ovarian Neoplasms/genetics , Protein Phosphatase 2/genetics , Cell Line, Tumor , Cell Proliferation , Female , Gene Expression Regulation, Neoplastic , Humans , Real-Time Polymerase Chain Reaction
14.
Nan Fang Yi Ke Da Xue Xue Bao ; 35(1): 77-82, 2015 Jan.
Article in Chinese | MEDLINE | ID: mdl-25613614

ABSTRACT

OBJECTIVE: To detect changes of Foxp3 expression in the decidua in patients with preeclampsia and investigate the correlation of Foxp3-924 (rs2232365) polymorphisms with preeclampsia. METHODS: From October 2011 to December 2012, 252 normal pregnant women and 156 preeclampsia patients of Han nationality from the same geographic region were tested for Foxp3-924 genotypes by polymerase chain reaction with sequence-specific primer (PCR-SSP). Sixty-eight of the patients with preeclampsia (33 with mild and 35 with severe preeclampsia) and 30 of the normal pregnant women were also examined for Foxp3 expression in the decidua using immunohistochemical method. RESULTS: Foxp3 positive expression rates in the decidua was 51.52% in mild preeclampsia and 28.57% in severe preeclampsia cases, significantly lower than that in the control group (86.67%, P<0.05). In preeclampsia patients, the frequencies of Foxp3-924G/G, G/A, and A/A genotypes were 0.1346, 0.4615 and 0.4038, respectively, and the frequencies of Foxp3-924A and Foxp3-924 G were 0.6346 and 0.3654, respectively. The genotype frequencies of Foxp3-924G/G, G/A and A/A in the control group were 0.1508, 0.4087 and 0.4405, respectively, and the frequencies of Foxp3-924 A and Foxp3-924 G were 0.6448 and 0.3552, respectively. No significant differences were found in the gene frequencies of Foxp3-924G/A between preeclampsia patients and the control group (P>0.05). CONCLUSION: The expression level of Foxp3 in the placental tissue of preeclampsia patients is significantly lower than that in normal pregnant women, suggesting that lowered Foxp3 expression decreases the immunosuppressive function and causes imbalance of immune tolerance between maternal-fetal to induce preeclampsia. Foxp3-924 polymorphisms is not significantly correlated with the occurrence of preeclampsia.


Subject(s)
Forkhead Transcription Factors/genetics , Placenta/metabolism , Polymorphism, Genetic , Pre-Eclampsia/genetics , Case-Control Studies , Female , Forkhead Transcription Factors/metabolism , Gene Frequency , Genotype , Humans , Pregnancy
15.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(5): 949-52, 2010 May.
Article in Chinese | MEDLINE | ID: mdl-20501364

ABSTRACT

OBJECTIVE: To establish immortalized B lymphoblast cell lines (B-LCLs) from healthy anti-HBs antibody (anti-HBs)-positive volunteers and screen for human anti-HBs and the antibody-secreting cells. METHODS: The peripheral blood mononuclear cells (PBMC) isolated from 3 healthy volunteers positive for anti-HBs with hepatitis B vaccine boost vaccination were infected with Epstein-Barr virus (EBV) and incubated in the presence of CpG DNA motifs and cyclosporin A (CyA). The anti-HBs in the culture supernatant of the immortalized B-cells was quantified by Architect anti-HBs assay with chemiluminescent microparticle technique. Immunocytochemistry was performed to identify the differentiation of the cell clones expressing anti-HBs. RESULTS: Immortalized B-cell culture was successfully established from the cell clones secreting anti-HBs with EBV infection and CpG DNA stimulation. The titer of anti-HBs in the culture supernatant was at its peak at 3 weeks of cell culture and then decreased gradually. At 3 months of cell culture, the cells still retained the capacity of anti-HBs production as verified by the results of immunocytochemistry for CD20 and CD138. CONCLUSION: Immortalized B-cell culture secreting anti-HBs from volunteers receiving boost hepatitis B vaccination has been successfully established by modified EBV immortalization technique.


Subject(s)
B-Lymphocytes/immunology , Cell Transformation, Viral , Hepatitis B Antibodies/immunology , Hepatitis B Vaccines/immunology , Immunization, Secondary , Cell Line , Hepatitis B/prevention & control , Hepatitis B Surface Antigens/immunology , Herpesvirus 4, Human/immunology , Humans , Vaccination
16.
Langmuir ; 26(8): 6009-14, 2010 Apr 20.
Article in English | MEDLINE | ID: mdl-20337480

ABSTRACT

Air-stable magnetic cobalt nanocrystals have been conveniently prepared via a reverse micellar synthesis, followed by a hydrothermal treatment. The synthesis was carried out by first mixing an aqueous solution containing cobalt chloride and poly(sodium 4-styrenesulfonate) (PSS) with an organic mixture containing cetyltrimethylammonium bromide (CTAB) to form reverse micelles, followed by reducing cobalt ions with sodium borohydride. The resultant nanoparticles were then undergone a hydrothermal treatment at 165 degrees C for 8 h to generate well-dispersed CTAB/PSS-encapsulated cobalt nanocrystals with an average diameter of 3.5 +/- 0.5 nm. The nanoparticles were highly crystalline with a hexagonal close-packed crystal phase. The presence of CTAB/PSS complex coatings was identified by FT-IR and UV-vis spectroscopies as well as thermogravimetry analyses. The nanocrystals exhibited superparamagnetic property at room temperature with a saturation magnetization (M(s)) of 95 emu/g. The magnetization could be largely preserved after storage at room temperature for 4 months as the M(s) value only slightly decreased to 88 emu/g (measured at 300 K). Thus, the polymer encapsulation could not only improve thermal stability of the micelles for the growth and nucleation of Co atoms but also protect the resulting cobalt nanocrystals from oxidation through forming an oxygen impermeable sheath.


Subject(s)
Cobalt/chemistry , Nanoparticles/chemistry , Polymers/chemistry , Surface-Active Agents/chemistry , Cetrimonium , Cetrimonium Compounds/chemistry , Metal Nanoparticles/chemistry , Nanotechnology/methods
18.
Zhonghua Yi Xue Za Zhi ; 89(14): 970-2, 2009 Apr 14.
Article in Chinese | MEDLINE | ID: mdl-19671309

ABSTRACT

OBJECTIVE: To compare the effects of sperm chromatin dispersion (SCD) test and TdT-mediated dUTP nick end labeling (TUNEL) assay in assessing the DNA fragmentation in human sperm. METHODS: Motile sperms were isolated from the semen samples obtained from 20 healthy fertile men and 32 clinically infertile patients by swim-up technique, and underwent SCD and TUNEL to analyze the DNA fragmentation. RESULTS: The rate of sperm with DNA damage of the infertile patients was 12.8% +/- 5.8% tested by SCD, significantly higher than that of the healthy fertile men (7.6% +/- 3.3%, t = 3.576, P = 0.001), and the rate of sperm with DNA damage of the infertile patients was 11.1% +/- 5.1% tested by TUNEL assay, significantly higher than that of the healthy fertile men (6.8% +/- 2.8%, t = 3.467, P = 0.001). The proportion of sperm cell with abnormal DNA integrity measured by SCD test was correlated strongly with that determined by TUNEL for the infertile men (r = 0.841, P = 0.000) and for the fertile men too (r = 0.823, P = 0.000). The rate of sperm with DNA damage measured by SCD were not significantly different from those of TUNEL-positive sperm in fertile men (t = 1.996, P = 0.060). The rate of sperm with DNA damage measured by SCD was significantly higher than that measured by TUNEL among infertile patients (t = 3.023, P = 0.005). CONCLUSION: The presence of sperm DNA damage may lead to male infertility. SCD is simpler, cheaper and more reliable than TUNEL in testing the sperm DNA damage.


Subject(s)
DNA Damage , DNA/genetics , Infertility, Male/diagnosis , Infertility, Male/genetics , Spermatozoa/chemistry , Adult , Case-Control Studies , DNA Fragmentation , Humans , Infertility, Male/metabolism , Male , Sperm Motility
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 28(6): 979-81, 2008 Jun.
Article in Chinese | MEDLINE | ID: mdl-18583243

ABSTRACT

OBJECTIVE: To explore the role of interferon (IFN)-alpha/beta receptor beta subunit (IFNAR2) in the patients' response to IFN-alpha therapy as influenced by the grade of chronic hepatic inflammation, and understand the relation of IFNAR2 expression in the peripheral blood mononuclear cells (PBMCs) with HBV infection. METHODS: Liver tissue specimens were obtained from 21 patients with chronic hepatitis B for examination of the hepatic inflammation, and PBMCs were isolated from another 16 patients with chronic hepatitis B and 15 health control subjects. Both the hepatic tissues and PBMCs were examined for IFNAR2 expression using immunohistochemistry. RESULTS: The 21 patients with chronic hepatitis B were divided into 3 groups according to the severity of hepatic inflammation, namely G(1) (n=3), G(2) (n=7) and G(3) (n=11) groups. The patients in G(3) group showed had significantly higher IFNAR2 expressions in liver (25.1307-/+7.0700) than those of the G(1) (5.6913-/+1.8422) and G(2) (7.4706-/+5.3572) groups (P=0.000). The IFNAR2 levels in the PBMCs, however, did not show significant difference between patients with chronic hepatitis B and the healthy control subjects. CONCLUSION: In patients with chronic hepatitis B, IFNAR2 expression level is positively correlated to the severity of hepatic inflammation, and increased IFNAR2 expression in severe hepatic inflammation is therefore likely to result in increased response rate to INF-alpha therapy. The expression of IFNAR2 in the PBMCs is not associated with HBV infection.


Subject(s)
Hepatitis B, Chronic/metabolism , Leukocytes, Mononuclear/metabolism , Liver/metabolism , Receptor, Interferon alpha-beta/metabolism , Female , Hepatitis B, Chronic/pathology , Humans , Immunohistochemistry , Liver/pathology , Male , Receptor, Interferon alpha-beta/blood
20.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 23(2): 165-8, 2006 Apr.
Article in Chinese | MEDLINE | ID: mdl-16604487

ABSTRACT

OBJECTIVE: To investigate and clarify whether the genetic susceptibility to women with hypertensive disorder complicating pregnancy or pre-eclampsia is associated with polymorphisms and couple sharing rate of transporter associate with antigen processing genes(TAP). METHODS: One hundred and two severe pre-eclampsia women and their spouses served as study group, and 200 normal pregnant women and their spouses were selected as control group. All pregnant women were primipara with single fetus. Genomic DNA was extracted from 2 mL cubital venous blood. We used the amplification refractory mutation system polymerase chain reaction(ARMS-PCR) to characterize TAP gene locus 333, 637, 379, 565, 665. RESULTS: We observed eleven TAP haplotypes. There were four kinds of haplotypes(1A-1D) existing in TAP1, and seven kinds of haplotypes(2A-2G) existing in TAP2. The gene frequencies of TAP2B(Chi2=9.19, P<0.01, RR=4.18) and TAP2F(Chi2=5.34, P<0.05, RR=4.63) of patient group with pre-eclampsia were significantly higher as compared with control group. The analyses of some TAP haplotypes such as TAP1B(Chi2=4.87, P<0.05, RR=3.14), TAP1C(Chi2=5.42, P<0.05, RR=4.90), TAP2B(Chi2=9.65, P<0.01, RR=5.39) showed that the couple sharing rate of pre-eclampsia women and their spouses had statistically a highly significant increase in comparison with that of controls. CONCLUSION: Our data suggest that the presence of TAP2B or TAP2F haplotypes should be considered as a risk increased to pregnant women being susceptible to hypertensive disorder complicating pregnancy; and also the elevated couple sharing rates of TAP1B, TAP1C and TAP2B genes will increase the opportunity or possibility of pregnant women suffering from pre-eclampsia disease.


Subject(s)
ATP-Binding Cassette Transporters/genetics , Hypertension/complications , Polymorphism, Genetic , Pregnancy Complications/etiology , Pregnancy Complications/genetics , Adult , Family Characteristics/ethnology , Female , Genotype , Humans , Male , Pregnancy
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