Objective To understand the running status and distribution of water-improving pmjects in Anda city.Methods The running and management of the water-improving projects were investigated and located the position by Global Positioning Systerm(GPS)in Anda city.Results Among 317 water-improving projects,16.09%were either long-term projects or in poor management or had already stopped usage,77 projeets were broken,accounting for 24.29%;all inadequate supply of equipment and pipeline,83 projects had never been started,accounting for 26.18%;now,106 projects were running,accounting for 33.44%.In only 46 projects,the water fluoride concentration was lower than 1.00 mg/L,accounting for 14.51%,as a result 36 thousand people benefited.Conclusions The running status of water-improving projects was unacceptable,most of them stopped running and endemic fluorosis control was still severe in Anda city.

Objective To study the inducement of U251 glioblastoma cell apoptosis in vivo through up-regulating PUMA expresion and knocking down miR-221/222, and explore its mechanism.Methods Nude mouse xenograft models were established in 5-week-old BALB/c nude mice by subcutaneous vaccination of U251 glioblastomas; 1 week later, they were treated with intratumoral injection of lipofcctamine-mediated miRNA-221/222 antisense oligonucleotides (GroupA), nonsense sequences (Group B) and controls (Group C),respectively (n=8).The tumor growth was monitored until the end of observation period (28 d after the treatment) and pathological changes of the glioblastoma tissues were observed by HE staining at the end of observation.Fluorescence in situ hybridization (FISH) and real-time PCR were employed to measure the miR-221 and miR-222 expressions. Terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling (TUNEL) assay was used to detect the apoptosis of glioblastomas.Immunohistochemistry and Westem blotting were used to detect the expressions of PUMA,bax,bcl-2 and p53 in removed tumor specimens. Results The volume in Group A was significantly smaller than that of those in group B and group C 6-28 dater treatment (P=0.006). The miR-221 and miR-222 mRNA expressions in Group A were significantly decreased as compared with those of those in group B and group C.HE staining indicated that decreased heteromorphism and reduced number of new vessels in Group A were noted as compared with those in group B and group C.The cell apoptotic index in Group A was significantly higher than that in group B and group C (P＜0.05).Immunohistochemistry showed that the expression levels of PUMA and bax in Group A was significantly up-regulated as compared with those in group B and group C, while the expression of bcl-2 in Group A was significantly down-regulated as compared with that in group B and group C; and no significant changes were noted in the p53 expression. Conclusion By up-regulating PUMA expresion,knocking down miR-221/222 can induce U251 glioma apoptosis in vivo.

Astrocytoma ; genetics ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Frizzled Receptors ; genetics ; metabolism ; Glioblastoma ; genetics ; metabolism ; Glioma ; genetics ; metabolism ; Humans ; Paraffin Embedding ; RNA, Messenger ; metabolism ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Signal Transduction ; Wnt2 Protein ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

OBJECTIVETo study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.

METHODSC6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.

RESULTSCx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.

CONCLUSIONThe mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.

Animals ; Apoptosis ; Cell Division ; physiology ; Connexin 43 ; genetics ; physiology ; DNA, Complementary ; genetics ; Glioma ; pathology ; Rats ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo detect the expression and the CpG island methylation status of tumor suppressor gene p15 after exposure to 1,4-benzoquinone (1,4-BQ) in primary cultivated C57BL/6J mouse bone marrow cells in vitro.

METHODSThe mouse bone marrow cells were isolated in vitro. The effect of 0, 0.1, 1, 5, 10, 20, and 40 µmol/L 1,4-BQ on cell viability (CKK-8) was detected. 0, 0.1, 1, 10 µmol/L 1,4-BQ were used to intoxicate the mouse bone marrow cells for 24 h; Real-time PCR was employed to analyze the mRNA expression level of p15; The bisulfite sequencing PCR (BSP) was used to look into the methylation status of CpG islands in p15 promoter region.

RESULTS1,4-BQ exhibited dose-dependent toxicity to mouse bone marrow cells, and the LC(50) was 8.3 µmol/L (95%CI: 4.6 - 10.6 µmol/L). The mRNA expression of p15 in 10 µmol/L group was only equivalent to 43% of control group. Compared with control group, the decrease of p15 mRNA expression in1 and 10 µmol/L concentration were obvious, and the differences had statistical significance (P < 0.05 or P < 0.01). BSP sequencing results were same between the exposure groups and control group, the 56 CpG sites on CpG islands remained in the state of unmethylated.

CONCLUSIONmRNA expression of p15 gene decreases after exposure to 1,4-BQ, but the CpG islands methylation status in promoter is not affected, suggesting that methylation does not participate in 1,4-BQ-mediated p15 gene expression decrease, other effect mechanisms still need to be investigated.

Animals ; Base Sequence ; Benzoquinones ; toxicity ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; CpG Islands ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; DNA Methylation ; Environmental Exposure ; Mice ; Mice, Inbred C57BL ; Promoter Regions, Genetic ; RNA, Messenger ; genetics

Objective To study the mechanism of miR-21 in regulating the invasion of human glioblastoma (GBM) cells in vitro. Methods The transfection reagent oligofectamine was mixed with antisense miRNA-21 (AS-miR-21) and nonsense oligodeoxyribonucleotides (ODN), respectively, and then, they were added into the medium of U251 GBM cell line as AS-miR-21 treatment group and nonsense ODN treatment group, respectively; control group (treated with PBS) was also established.MiR-21 luciferase reporter assay was used to detect the miR-21 knocking down effect. Matrigel cell growth assay and Transwell assay were used to determine the invasion and migration abilities of U251 cells. Western blotting was employed to test the expressions of invasion-related proteins (FAK,MMP-9/2, TIMP-1 and Tubulin-α); immunofluorescence was also employed to observe the morphology of Tubulin-α protein in GBM cells. Results Luciferase intensity in as-miR-21 treated U251 cells was significantly suppressed as compared with that in the control group and nonsense ODN treatment group (P＜0.05). The diameter of cultured clone in as-miR-21 treated U251 cells was smaller than that in the controls and nonsense ODN treatment group (F=102.819, P=0.000). Decreased cells via the transwell member in thc AS-miR-21 treatment group were detected as compared with those in the controls and cnonsensc ODN treatment group (F=243.465, P=0.000). The expressions of FAK, MMP-2/9 weredown-regulated and that of TIMP-1 was up-regulated in the AS-miR-21 treated tumor cells as compared with the other 2 groups (P＜0.05). No obvious changes were noted on the expression of Tubulin α,however, the morphology of Tubulin α protein in the AS-miR-21 treatment group changed. Conclusion High expression of miR-21 induce the ability of U251 GBM cell invasion and miR-21 can be taken as a candidate for gene therapy of human glioma.

Objective To explore the effects of nursing intervention on school-age children with asthma received the treatment of oxygen-driven inhalation.Methods Totals of 167 cases were randomly divided into intervention group (84 cases) and control group (83 cases).All children received conventional treatment and care,on this basis,observation group received audio-visual training,behavioral training and oxygenation therapy nursing intervention during oxygen inhalation therapy,while control group received health education according to the traditional oxygen inhalation therapy,and inhalation therapy were completed by patients.Then,treatment effects of two groups were observed and compared.Results In intervention group,after inhalation,respiration was( 18.31 ± 2.32)cpm,heart rate was (91.23 ± 2.65 )cpm and blood oxygen saturation index was (97.25 ±2.03) percent,and that in control group was ( 21.27 ± 3.08 ) cpm,(95.87 ± 3.42) cpm and ( 95.02 ± 3.17 )percent,difference between two groups was statistically significant( t =7.020,9.807,5.420,respectively ; P ＜ 0.01 ).Intervention group asthma symptom score of day and night was (1.05 ± 0.23 )and (1.03 ± 0.21 )lower than control group that was ( 1.77 ± 0.35 ) and ( 1.75 ± 0.32 ),the difference between two groups was statistically significant (P ＜ 0.01 ).Intervention group inhalation therapy and treatment compliance were significantly better than control group,and the difference was statistically significant (x2 =34.53,10.92,respectively; P ＜0.01 ).Conclusions Whole process nursing intervention can improve therapy compliance of children,quickly alleviate disease condition,thereby improve the effects of inhalation treatment.

OBJECTIVETo study the inhibitory effect of knocking down microRNA(miR)-221 and miR-222 on human glioma cell growth and its possible mechanism.

METHODSmiRNA-221/222 antisense oligonucleotides (antisense miR221/222) were transfected into human glioma U251 cells by lipofectamine. Northern blot analysis was conducted to detect the mRNA expression of miR-221/222 in the control and transfected cell groups. The proliferation activity of cells was determined by MTT assay. Cell invasion ability was examined by transwell assay, and cell cycle kinetics and apoptosis were detected with flow cytometry. The expression of relevant proteins was analyzed by Western blotting. The therapeutic efficacy of antisense miR221/222 on the growth of xenograft tumors in nude mice were also observed.

RESULTSIn the antisense miR-221/222-transfected cells, the expression of miR-221/222 was significantly reduced; the cell invasion ability was suppressed, cell cycle was blocked at G(0)/G(1) phase, and apoptotic cells were increased. The growth of xenograft tumors treated with antisense miR-221/222 was also inhibited. In antisense miR-221/222 treated tumor cells, the expression of bcl-2 was down-regulated while connexin43, p27, PUMA, caspase-3, PTEN, TIMP3 and Bax up-regulated, and p53 expression not changed.

CONCLUSIONThere is a significant inhibitory effect of antisense miR-221/222 on the growth of human glioma U251 cells. miR-221/222 may be considered as a candidate target for gene therapy of human gliomas.

Animals ; Apoptosis ; Base Sequence ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Genetic Therapy ; Glioma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; MicroRNAs ; biosynthesis ; genetics ; Molecular Sequence Data ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-3 ; metabolism ; Transfection

OBJECTIVETo investigate the differential gene expression of ependymomas.

METHODSFour fresh samples of ependymomas and 1 of normal brain tissue were collected during operation. The extracted total RNAs were converted as (32)P tagged cDNA probes, which were then hybridized with the Atlas Human Cancer Array, producing the array based hybridization maps following the protocol provided with the kit. A set of special software was applied to the analysis and RT-PCR was performed to test the result.

RESULTIn comparison with the normal brain tissue, there were 31 upregulated gene and 1 downregulated gene in ependymomas, most of which were firstly found to be differentially expressed in this kind of tumor.

CONCLUSIONThe discrepancy of gene expression profiles between ependymomas and normal brain tissues is highly put through and effectively detected with cDNA array, which provides new information for the further research on the molecular mechanisms of this lesion.

Brain ; metabolism ; Brain Neoplasms ; genetics ; Ependymoma ; genetics ; Gene Expression Profiling ; Humans ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND:Internal and external fixation combined with autologous bone graft for treating atrophic nonunion has a long treatment cycle,and moreover,it cannot achieve a 100％ cure rate.Platelet-rich plasma contains a variety of growth factors and a large number of white blood cells,and contributes to tissue healing.However,there is no clinical study on the effectiveness of platelet-rich plasma combined with conventional surgery in the treatment of atrophic nonunion.OBJECTIVE:To investigate the effectiveness of platelet-rich plasma in the treatment of atrophic nonunion of femoral shaft fractures.METHODS:We conducted a prospective,open-label,randomized,controlled clinical trial at the Affiliated Hospital of Qinghai University,China.Ninety-two patients with atrophic nonunion of femoral shaft fractures were equally and randomly divided into control group and experimental group.Patients in the control group received conventional surgery.Patients in the experimental group were injected with autologous platelet-rich plasma on the basis of conventional surgery.The primary outcome was fracture healing rate at postoperative 9 months.The secondary outcomes were visual analogue scale scores in resting state and during passive motion,healing time,treatment costs,and adverse reactions.The study protocol was approved by the Ethics Committee of Affiliated Hospital of Qinghai University of China (approval number:QHG0223A) on May 20,2014.Written informed consent was provided by each patient and their family members after they fully understood the treatment plan.RESULTS AND CONCLUSION:Our partial results demonstrated that visual analogue scale scores and complications were similar between the two groups at postoperative 1-3 days.The healing rate was significantly higher in the experimental group than in the control group.The healing time was significantly shorter in the experimental group than in the control group.This trial will provide objective data for the clinical use of platelet-rich plasma combined with conventional surgery for the treatment of atrophic nonunion.