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Objective@#To investigate the awareness of air pollution protection knowledge and its influencing factors among primary school students in Shennongjia Forest District, Hubei Province, so as to provide insights into targeting implementation of health education on air pollution protection knowledge.@*Methods@#Students in Grade 3 to 5 in Shennongjia Shiyan primary school were enrolled by stratified cluster sampling method, and students' demographic features and awareness of air pollution protection knowledge were investigated using the Investigation on the Effects of Air Pollution Health Protection of Pupils (Volume A). Factors affecting the awareness of air pollution protection knowledge among primary school students were identified using a multivariable logistic regression model.@*Results@#A total of 897 questionnaires were allocated, and 877 valid questionnaires were recovered, with an effective rate of 97.77%. The respondents included 446 men (50.86%) and 431 women (49.14%), 301 third grade students (34.32%), 284 fourth grade students (32.38%), and 292 fifth grade students (33.30%), and had a mean age of (10.32±0.93) years. The overall awareness of air pollution protection was 55.76%, and the awareness rates of basic concepts, basic knowledge, and basic behaviors and skills were 42.99%, 53.48% and 57.24%, respectively. Multivariable logistic regression analysis identified age (OR=1.453, 95%CI: 1.053-2.005), living with parents (OR=2.638, 95%CI: 1.571-4.429), mother's educational level (below primary school, OR=0.270, 95%CI: 0.084-0.862; primary school, OR=0.169, 95%CI: 0.069-0.416; junior high school, OR=0.309, 95%CI: 0.138-0.691; high school, OR=0.352, 95%CI: 0.160-0.773) and average annual family income (50 000 to 100 000 Yuan, OR=1.629, 95%CI: 1.162-2.282; 100 000 to 150 000 Yuan, OR=1.802, 95%CI: 1.101-2.948; ≥150 000 Yuan, OR=1.939, 95%CI: 1.065-3.529) as factors affecting the awareness of air pollution protection knowledge among primary school students.@*Conclusion@#The awareness of air pollution protection knowledge is 55.76% among primary school students in Shennongjia Forest District, and is affected by age, mother's educational level, average annual family income and living with parents.
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@# 树突状细胞(DC)是体内功能强大的抗原提呈细胞(APC),在机体抗肿瘤免疫反应的过程中起着关键的作用。成熟 DC具有激活T淋巴细胞并激活抗肿瘤免疫反应的功能, 以DC为基础的抗肿瘤免疫疗法显示出良好的应用前景。免疫检查点疗 法是肿瘤免疫治疗的另一强有力手段, 以PD-1和CTLA-4为代表的免疫检查点分子在肿瘤微环境中起着免疫调节的作用,同时 也对DC的成熟和功能起着重要的调控作用。肿瘤微环境中的未成熟DC和免疫检查点分子是肿瘤免疫逃逸的重要因素。因此 探究免疫检查点分子对DC成熟及功能的调控机制对于抗肿瘤治疗的研究具有非常重要的意义。本文从DC的视角,阐述了肿瘤 微环境中免疫检查点分子对DC成熟及功能的调控机制以及免疫检查点靶向药物联合DC疫苗应用于肿瘤临床实验的最新研究 进展。
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Objective @#To investigate the morphology and proliferation viability in oxidative stress induced damage in human MG63 cells. @*Methods@# The MG63 cells were treated with superoxide anion (O2.') produced by different concentrations of xanthine/xanthine oxidase enzymatic reactions to establish the model of oxidative stress in MG63 cells, using the xanthine oxidase inhibitor oxypurinol to observe the reverse effect of oxypurinol on xanthine/xanthine oxidase induced damage in human MG63 cells. Using the flow cytometry, the production of intracellular reactive oxygen species (ROS) induced by xanthine/xanthine oxidase induced cellular oxidative stress damage was evaluated by the oxidation⁃sensitive fluorescent probe, the 2’7' dichlorofluorescin diacetate. Cellular viability and morphology was evaluated by the MTT assay and the phase contrast microscope.@*Results @#Xanthine/xanthine oxidase induced intracellular ROS production in a dose and time dependent manner (P < 0.05). The cellular viability was reduced and cellular morphology was damaged, too (P < 0.05). Xanthine/xanthine oxidase induced the damage of the cellular morphology. At the same processing time, the higher the xanthine/xanthine oxidase concentration, the higher intracellular ROS fluorescence intensity value, and the lower OD value, the difference was statistically significant (P < 0.05). The intracellular mean ROS fluorescence intensity in xanthine/xanthine oxidase + oxypurinol combined treatment group was significantly lower compared with the same concentration of xanthine/xanthine oxidase (P < 0.05). At the same concentration of xanthine/xanthine oxidase, with the extension of treatment time, the intracellular mean ROS fluorescence intensity gradually increased, the OD value decreased, compared with the control group, the intracellular mean ROS fluorescence intensity of 120 min increased to 345% of the control, was the highest among the xanthine/xanthine oxidase groups. The OD value of 24 h was the 22.9% of the control group, was the lowest among the xanthine/xanthine oxidase groups, cell proliferation activity decreased more obvious. @*Conclusions@#Xanthine/xanthine oxidase could induce oxidative stress damaged the cellular morphology and reduced the cellular viability in MG63 cell lines. The oxypurinol (the inhibitor of xanthine oxidas) could reverse the oxidative stress injury induced by xanthine/xanthine oxidase in human osteoblastic cells.
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9α-hydroxy-4-androstene-3,17-dione (9-OH-AD) is an important intermediate in the steroidal drugs production. 3-ketosteroid-9α-hydroxylase (KSH), a two protein system of KshA and KshB, is a key-enzyme in the microbial steroid ring B-opening pathway. KSH catalyzes the transformation of 4-androstene-3,17-dione (AD) into 9-OH-AD specifically. In the present study, the putative KshA and KshB genes were cloned from Mycobacterium smegmatis mc(2)155 and Gordonia neofelifaecis NRRL B-59395 respectively, and were inserted into the expression vector pNIT, the co-expression plasmids of kshA-kshB were obtained and electroporated into Mycobacterium sp. NRRL B-3805 cells. The recombinants were used to transform steroids, the main product was characterized as 9α-hydroxy-4-androstene-3,17-dione (9-OH-AD), showing that kshA and kshB were expressed successfully. Different from the original strain Mycobacterium sp. NRRL B-3805 that accumulates 4-androstene-3,17-dione, the recombinants accumulates 9α-hydroxy-4-androstene-3,17-dione as the main product. This results indicates that the putative genes kshA, kshB encode active KshA and KshB, respectively. The process of biotransformation was investigated and the results show that phytosterol is the most suitable substrate for biotransformation, kshA and kshB from M. smegmatis mc(2)155 seemed to exhibit high activity, because the resultant recombinant of them catalyzed the biotransformation of phytosterol to 9-OH-AD in a percent conversion of 90%, which was much higher than that of G. neofelifaecis NRRL B-59395. This study on the manipulation of the ksh genes in Mycobacterium sp. NRRL B-3805 provides a new pathway for producing steroid medicines.
Subject(s)
Androstenedione , Bacterial Proteins , Genetics , Metabolism , Biotransformation , Ketosteroids , Mixed Function Oxygenases , Genetics , Metabolism , Mycobacterium , Metabolism , Mycobacterium smegmatis , PlasmidsABSTRACT
<p><b>OBJECTIVE</b>To study the expression of WAVE1 and p22phox in peripheral blood mononuclear cells (PBMCs) in children with acute lymphocytic leukemia (ALL) and the relationship of WAVE1 with oxidative stress.</p><p><b>METHODS</b>Real-time PCR was used for detecting WAVE1 and p22phox expression in PBMCs in 41 children with ALL and 10 normal controls. Plasma activity of superoxide dismutase (SOD) was measured by the xanthine oxidase method. Plasma activity of GSH-Px was measured by the DTNB reaction test.</p><p><b>RESULTS</b>The expression of WAVE1 and p22phox was significantly higher in the active ALL groups (newly diagnosed and relapse ALL) than that in the normal control and the complete remission (CR) ALL groups (<0.01). The CR ALL group showed increased WAVE1 and p22phox expression than those in the normal control group (<0.05). Plasma activities of SOD (22.62+/-7.39 U/mL) and GSH-Px (91.73+/-28.88 micromol/L) in the active ALL group were significantly lower than those in the normal control (166.35+/-27.93 U/mL and 490.94+/-39.38 micromol/L, respectively) and the CR ALL groups (107.11+/-28.57 U/mL and 267.56+/-82.64 micromol/L, respectively) (<0.01). WAVE1 expression was positively correlated with p22phox expression (r=0.34, <0.05) but negatively correlated with plasma activities of SOD and GSH-Px ( r=-0.336 and-0.408, respectively; <0.05).</p><p><b>CONCLUSIONS</b>WAVE1 and p22phox expression in PBMCs increased and was associated with the disease course in children with ALL. Oxidative stress may be involved in the regulation of WAVE1 expression in ALL children.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Glutathione Peroxidase , Blood , Leukocytes, Mononuclear , Metabolism , NADPH Oxidases , Genetics , Oxidative Stress , Polymerase Chain Reaction , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Metabolism , RNA, Messenger , Blood , Superoxide Dismutase , Blood , Wiskott-Aldrich Syndrome Protein Family , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate role of WASP family verprolin homologous protein 1 (WAVE1) in K562 leukemia cell invasion and its mechanism.</p><p><b>METHODS</b>Immunofluorescence method was used to detect the distribution of WAVE1 and MMP-2 in the cells. K562 cells were transfected with pcDNA3. 1-WAVE1 reconstructed plasmid or with specific siRNA to WAVE1 gene. The invasion ability of K562 cells was examined by Transwell assay. The expression level of WAVE1 and MMP-2 in K562 cells was assayed by real-time PCR and Western blot.</p><p><b>RESULTS</b>(1) WAVE1 and MMP-2 mainly expressed and co-localized in the cell membrane; (2) 24 h and 48 h after transfected with pcDNA3. 1-WAVE1, the MMP-2 mRNA level in K562 cells increased by 295% and 198% while its protein increased by 80% and 23% respectively as compared with control K562 cells. At the same time point after transfected with specific siRNA, the MMP-2 mRNA level decreased by 81% and 28%, and its protein decreased by 36% and 53% respectively as compared with control. (3) The invasion ability of K562 cells was enhanced after transfected with pcDNA3. 1-WAVE1 and depressed after transfected with the specific siRNA.</p><p><b>CONCLUSION</b>The co-localization of WAVE1 and MMP-2 in K562 cells suggests they coordinate in functions; WAVE1 may involve in the migration and invasion of K562 cells through regulating the expression level of MMP-2.</p>
Subject(s)
Humans , K562 Cells , Leukemic Infiltration , Genetics , Metabolism , Matrix Metalloproteinase 2 , Genetics , Metabolism , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Transfection , Wiskott-Aldrich Syndrome Protein Family , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the therapeutic effect and the influencing factors of event-free survival (EFS) of childhood acute lymphoblastic leukemia (ALL) in Xiangya Hospital of Central South University and the First Affiliated Hospital of Guangxi Medical University.</p><p><b>METHODS</b>All the patients adopted chemotherapy according to therapeutic guideline revised by the Subspecialty Group of Hematology, The Society of Pediatrics, Chinese Medical Association for the second-time in 1998 (the Rongcheng ALL-98 Protocol). Kaplan-Meier method was used to estimate the survival rates of 188 patients who received therapy with good compliance. The differences of EFS between groups were assessed by Log-rank test. The independent influencing factors on EFS were analyzed by the Cox proportional hazards regression model.</p><p><b>RESULTS</b>After receiving inductive treatment, 354 of 374 (93.6%) patients demonstrated a complete remission; 188 patients who received complete courses of treatment with good compliance showed (68.1 +/- 5.6)% five-year EFS. Meanwhile, the five-year EFS in standard-risk (SR) group and high-risk (HR) group were (75.2 +/- 6.0)% and (47.6 +/- 11.6)%, respectively. The total relapse rate was 10.6% and the median time to relapse was 13 months. Twenty-nine of 188 patients (15.4%) were dead, and 13 patients (7.0%) died from treatment-related complications. Independent adverse prognostic factors included risk grouping, t (9; 22)/bcr-abl gene and leukocyte count.</p><p><b>CONCLUSIONS</b>The total EFS of childhood ALL patients treated with Rongcheng ALL-98 Protocol in two hospitals was close to 70%. Therefore, it is necessary to evaluate risk factors and consider the grouping in more detail to reduce the treatment-related mortality and to increase the compliance of treatment which can ultimately improve the EFS.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , China , Epidemiology , Disease-Free Survival , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Drug Therapy , Epidemiology , Mortality , Prognosis , Survival Rate , Treatment OutcomeABSTRACT
The effects of Oxymatrine (Oxy) on the proliferation and apoptosis of human esophageal carcinoma Eca109 cell line and the mechanism were investigated. The human esophageal carcinoma Eca109 cells were cultured in vitro. The Oxy-induced apoptosis of Eca109 cells was assayed by using flow cytometry. The expressions of p-ERK1/2, Cyclin D1, p21waf/cip1, Bax and Bcl-2 were detected by Western blot. Flow cytometry revealed that Oxy could induce the apoptosis of Eca109 cells. Western blot showed that Oxy of different concentrations suppressed the expressions of p-ERK1/2, cyclin D1 and Bc1-2, but up-regulated the expression of p21waf/cip1 and Bax, and the ratio of Bax/Bcl-2 was increased It was suggested the Oxy could induce the apoptosis of Eca109-cells, which might be related to the upregulation of p21waf/cip1 and the downregulation of p-ERK1/2 Cyclin D1 and p21waf/cip1. The possible pathway may be related to Bcl-2/Bax.
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<p><b>OBJECTIVE</b>To investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>WAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.</p><p><b>RESULTS</b>WAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).</p><p><b>CONCLUSIONS</b>The WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Antibiotics, Antineoplastic , Pharmacology , Apoptosis , Blotting, Western , Cell Proliferation , Doxorubicin , Pharmacology , Jurkat Cells , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Metabolism , RNA, Messenger , Wiskott-Aldrich Syndrome Protein Family , Genetics , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of high mobility group boxl (HMGBI) gene silence on adriamycin (ADM)-induced apoptosis in K562/A02 drug resistance leukemia cells.</p><p><b>METHODS</b>K562/ A02 cells were transient transfected with HMGB1- small interference RNA(siRNA) vector, and the levels of HMGB1 gene differential expression pre-and post-transfection were measured by RT-PCR and Western blotting. 50% inhibition concentration (IC50) of ADM on K562/A02 was determined by WST-8 assay. Cell apoptosis was assessed by flow cytometry. The release of Smac/DIABLO from the mitochondria to the cytoplasm was assayed by Western blotting. Activity of Caspase-3 was assayed with a Caspase Colorimetric Assay Kit.</p><p><b>RESULTS</b>(1) The HMGB1 expression at mRNA and protein levels in HMGB1 siRNA transfected K562/A02 cells were decreased by 86% and 71% respectively compared with control. (2) Suppression of HMGB1 by siRNA in K562/A02 cells resulted in a reversal of the resistance to ADM, and decreased IC50 from (4.83 +/- 0.08) microg/ml to (1.33 +/- 0.10) microg/ml. 1 microg/ml and 5 microg/ml of ADM treatment increased cell apoptotic rate by 27% and 32% respectively. (3) HMGB1 suppression in K562/A02 cells significantly promoted ADM- induced Smac/DIABLO release from the mitochondria to the cytoplasm, and increased the activities of Caspase-3.</p><p><b>CONCLUSION</b>HMGB1 gene silence can enhance sensitivity of K562/A02 cells to ADM and reverse cell resistant to ADM.</p>
Subject(s)
Humans , Apoptosis , Genetics , Doxorubicin , Pharmacology , Gene Silencing , HMGB1 Protein , Genetics , K562 Cells , RNA, Small Interfering , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate if WAVE1 is involved in mult drug-resistance (MDR) of human leukemia cell line K562/A02.</p><p><b>METHODS</b>The level of WAVE1 in K562 and K562/A02 cells was assayed by Western blot and RT-PCR; K562 cells and K562/A02 cells were transient transfected with pEFBOS-WAVE1 reconstructed plasmid or specifically siRNA to WAVE1. 50% inhibition concentration (IC50) of doxorubicin on K562/A02 was determined by WST-8 assay. Hoechst33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. The mRNA level of mdrl was assayed by RT-PCR. The Bcl-2 protein was assayed by Western blot.</p><p><b>RESULTS</b>1. The WAVE1 expression at mRNA and protein level in K562/A02 cells was increased by about 70% and 63% respectively as compared with that in K562 cells. 2. Overexpression of WAVE1 in K562 cells by transient transfection significantly increased the resistance to doxorubicin, and increased IC50 from (0.05 +/- 0.00) microg/ml to (2.99 +/- 0.12) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was decreased by 30% or 35% respectively. 3. Suppression of WAVE1 in K562/A02 cells by siRNA resulted in a reversal of MDR to doxorubicin, and decreased IC50 from (4.29 +/- 0.15) microg/ml to (1.85 +/- 0.07) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was increased by 24% or 21% respectively. 4. Overexpression of WAVE1 in K562 cells significantly increased the mdrl mRNA and the Bcl-2 protein, and suppression of WAVE1 in K562/A02 cells by siRNA decreased the mRNA and the protein.</p><p><b>CONCLUSION</b>WAVE1 involves in the MDR mechanisms in K562/A02 leukemia cells through regulation the level of mdrl and Bcl-2.</p>
Subject(s)
Humans , Apoptosis , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Genetic Vectors , K562 Cells , Plasmids , Genetics , RNA Interference , RNA, Messenger , Genetics , Transfection , Wiskott-Aldrich Syndrome Protein Family , Genetics , MetabolismABSTRACT
Objective To study the characteristics of mesenchymal stem cells(MSCs)derived from chronic myelogenous leukemia(CML)patients' bone marrow and examine their hematopoiesis support in vitro.Methods MSCs from CML were obtained and cultured.Immunophenotype and in vitro differentiation capacities were investigated.Moreover,the Ph chromosome and bcr/abl gene of CML derived MSCs were detected.The expression of cytokines was detected by RT-PCR,and hema- topoiesis support of MSCs in vitro was detected by long-term bone marrow culture and the methylcel- lulose progenitor assay.Results CML derived MSCs showed a typical fibroblast like morphology. They were positive for CD29,CD44 and CD105,while negative for CD14,CD31,CD45,CD34 and HLA-DR.Under suitable conditions,CML derived MSCs could differentiate into osteoblasts and adi- pocytes.Further,CML derived MSCs showed a normal choromosome,and did not express bcr/abl gene.At last,they expressed hematopoiesis cytokines and possessed ability of hematopoiesis support in vitro.Conclusion There are MSCs with multidirectional differentiation potential in CML bone mar- row and they possess the ability of hematopoiesis support.