Background Choroidal neovascularization (CNV) is a serious complication of many fundus diseases.A variety of factors are associated with CNV.Research showed that recombinant human endostatin ( rhendostatin ) can arrest the vascular endothelial growth factor (VEGF) pathway and inhibit the proliferation of endothelial cells and angiogenesis.Objective This study was to observe the inhibition of rh-endostatin on experimental CNV.Methods The CNV animal models were created by Argon laser with the wavelength 532 nm to irradiate the inferior retina away optical disc 1-2 DD for 25 spots in 32 eyes of 16 chinchilla rabbits.The laser parameters were as follows:power 800 mW,spot diameter 75 μm and time shutter 50 ms.The models were then divided into model control group and rh-endostatin group.Rh-endostatin was intravitreously injected via scleral incision in 16 eyes of 8 model rabbits at 1 week after photocoagulation.Fundus fluorescein angiography (FFA) and optical coherence topography(OCT) were performed at 1,2,4 weeks after photocoagulation respectively.The eyeballs were enucleated and the retinal sections were prepared for the histopathologieal examination,and the contents of VEGF and pigment epithelial derived factor(PEDF) in rabbit vitreous,and blood serum were detected by ELISA at 2,4 weeks after photocoagulation.Results Retinal edema and exudes were seen in 1 week and scarring in 4 weeks after photocoagulation.In rh-endostatin injection group,the hyperfluorescence masses were seen in the background phase and early arterial phase in 42％ (84/200) of spots in the first week.The fluorescence leakage was decreased in the rh-endostatin injection group compared with control group in the second week and ceased at the third week on the FFA after injection.Variety forms of hyperreflective zones were found below the retinal pigment epithelium on the seventh day after photocoagulation.But the partial vessel occlusion and fibroplasias were identified in the rh-endostatin injection group in the third week by the OCT.The histopathological examination showed that the morphological abnormality was mild in the rh-endostatin injection group in comparison with model group.The serum PEDF concentration was significantly elevated but the VEGF/PEDF values in vitreous and serum were declined in rhendostatin injection group compared with model group (P ＜ 0.0 1 ).Conclusions Argon laser photocoagulation could induce the experimental CNV in chinchilla rabbit.Intravitreous injection of rh-endostar can effectively inhibit laser-induced CNV in rabbit.

Adult ; Benzamides ; pharmacology ; Humans ; Hypereosinophilic Syndrome ; drug therapy ; genetics ; Imatinib Mesylate ; Male ; Mutation ; Oncogene Proteins, Fusion ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; mRNA Cleavage and Polyadenylation Factors

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

Objective To discuss the level and its relativity analysis of IL-16 and IgE in asthmatic children .Methods Compare and analyze the level of IL-16,IL-10 and IgE between 18 asthmatic children and 14 healthy children.Results The serum level of IL-16 and IgE in asthmatic children was much higher than the contrast, P

OBJECTIVETo investigate changes in the adherent ability, the expression of adhesion related proteins Pyk2 and paxillin during HL-60 cells differentiation into granulocyte-monocyte induced by low-dose (LD) bufalin in combination with all-trans retinoic acid (ATRA), and to explore the effects of bortezomib on cellular adhesion and the expression of Pyk2 and paxillin.

METHODSThe expression of CD11b was detected by flow cytometry, cellular adherence ability by MTT assay, and the expressions of Pyk2, paxillin and tubulin by Western blot.

RESULTSThe combination of 5 nmol/L bufalin and 30 nmol/L ATRA induced HL-60 cells differentiation in a time-dependent manner, the percentages of CD11b positive cells treated for 2 d and 4 d being (20.0 +/- 2.8)% and (75.0 +/- 5.3)%, respectively, with the increasing of cellular adherence ability. Meanwhile the expressions of Pyk2 and Paxillin were also up-regulated in a time-dependent manner. Bortezomib suppressed HL-60 cell adhesion in a dose-dependent manner. At concentrations of 1 nmol/L and 10 nmol/L the adherence level were (7.8 +/- 0.1)% and (5.3 +/- 0.3)%, respectively, with down-regulation of Pyk2 but not Paxillin.

CONCLUSIONPyk2 is involved in the regulation of cellular adherence function. Bortezomib might inhibit HL-60 cells adhension function by down-regulation of Pyk2 expression.

Boronic Acids ; pharmacology ; Bortezomib ; Bufanolides ; pharmacology ; Cell Adhesion ; drug effects ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 2 ; metabolism ; HL-60 Cells ; Humans ; Paxillin ; metabolism ; Pyrazines ; pharmacology ; Tretinoin ; pharmacology

Cells, Cultured ; Chemokine CCL4 ; Chemotaxis, Leukocyte ; drug effects ; Endothelial Cells ; cytology ; metabolism ; Homocysteine ; pharmacology ; Humans ; Macrophage Inflammatory Proteins ; biosynthesis ; genetics ; Monocytes ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

Background and purpose:Remarkable advances have been made in cancer chemotherapy by developing new anticancer drugs and therapy strategies.However,multidrug resistance in human cancers remains a major clinical challenge for cancer treatment.Attempts in several clinical studies to reverse multidrug resistance protein (MDR) by using MDR modulators have not yet generated promising results.Our aim was to explored the mechanism of reversal of multidrug resistance in human leukemia K562 cells by PI3-K inhibitor.Methods:Trypanblue dye exclusion method was used to observe the drug sensitivity and the effect of LY294002 on the drug resistance.Western blot to analyze P-gp and p-Akt phenotypes,and flow cytometer was used to measure the intracellular drug accumulation. Results:K562/D induced by DNR was cross resistant to DNR,ADR,VCR and VP16 (Resistance Index:65,52,134 and 50 respectively).DNR induced over-expressions of P-gp and p-Akt in K562/D cells;LY294002 increased the intracellular drug accumulation,and then reversed the drug resistance to DNR,ADR,VCR and VPI6 in K562/D cells(Resistance Index:23,21,63 and 29 respectively),but not in the sensitive cells (K562/S).Conclusion:The multidrug resistance of K562/D cells can be induced by DNR which is related to the P-gp and p-Akt over-expressions, and LY294002 can reverse multidrug resistance in human leukemia cells in vitro via inhibits PI3-K/Akt pathway.

Objective To understand the status of hand hygiene(HH)of health care workers(HCWs)in a tertiary hospital in Zhuhai.Methods HH compliance of doctors,nurses,and cleaners randomly selected from 15 clinical de-partments in the whole hospital was observed through concealed observation by medical interns in October-Novem-ber 2016.Results HH compliance rate and correct rate of clinical departments in the whole hospital were 33.44%(1 131/3 382)and 59.86%(677/1 131)respectively,there was a significant difference in the compliance rate of HH among different types of HCWs (χ2 ＝12.610,P＝0.002),HH compliance rate from high to low was nurses (35.85%),cleaners (32.28%),and doctors (29.50%).Of five HH moments,HCWs’HH compliance rate after patient’s body fluid exposure was the highest (69.74%),while after touching patient surroundings was the worst (25.03%).HH compliance rates of HCWs with different occupations at different HH moments were all significant-ly different (all P＜0.05),HH compliance rates of doctors before aseptic procedure and after patient’s body fluid exposure were higher than nurses(71.25% vs 32.44%;82.86% vs 69.78%,respectively),HH compliance rate of nurses was highest after touching a patient(40.06%).Conclusion HH status in this hospital is not optimistic,HH compliance rate and correct rate are low,HH compliance rates of HCWs with different occupations and at different HH moments are both different,which need to be improved.

BACKGROUND AND OBJECTIVEc-Cbl and Cbl-b are two ubiquitous members of the Casitas B-lineage lymphoma (Cbl) family, which play important roles in the downregulation of epidermal growth factor receptor (EGFR) by acting as E3 ubiquitin ligases and multiadaptor proteins. This study investigated the expression of c-Cbl, Cbl-b, and EGFR in gastric carcinoma and its clinical significance.

METHODSThe expressions of c-Cbl, Cbl-b, and EGFR were detected by immunohistochemistry using tissue microarrays consisting of 124 specimens of gastric carcinoma and 16 specimens of normal gastric mucosa. The relationship between the expressions of c-Cbl, Cbl-b, and EGFR and clinicopathologic factors of gastric carcinoma were analyzed statistically.

RESULTSThe positive rates of c-Cbl, Cbl-b, and EGFR were higher in the gastric carcinoma group than in the normal group (71.0% vs. 18.0%, P<0.01; 82.3% vs. 25.0%, P<0.01; 56.5% vs. 12.5%, P<0.01, respectively). The expression of c-Cbl was positively correlated with depth of invasion (r=0.219, P=0.015), and TNM staging (r=0.266, P=0.003). The expression of Cbl-b was positively correlated with lymph node metastasis (r=0.190, P<0.034) and TNM staging (r=0.298, P<0.001). The expression of EGFR was positively correlated with depth of invasion (r=0.286, P<0.001) and TNM staging (r=0.362, P=0.000). The expression of both c-Cbl and Cbl-b was positively correlated with EGFR (r=0.241, P=0.007; r=0.183, P=0.042, respectively). Synchronous strong-positive expressions of c-Cbl, Cbl-b, and EGFR were observed in 27 specimens of gastric carcinoma, most of which were at advanced stage.

CONCLUSIONSOverexpressions of c-Cbl, Cbl-b, and EGFR are closely related to the invasion and progression of gastric carcinoma. c-Cbl and Cbl-b may serve as novel molecular markers for gastric carcinoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Disease Progression ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proto-Oncogene Proteins c-cbl ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Young Adult

Objective To study and discuss what part does methylated Id4 gene participate in malignant lymphoma stage Ⅳ by detecting the extent of how much Id4 gene has been methylated in afflicted children who suffer from malignant lymphoma.Methods Forty-two patients who had diagnosed with malignant lymphoma [Hodgkin's disease (HD),Non-Hodgkin's lymphoma(NHL)] were selected as study group.Their chemotherapy stages of pre-treatment,early-treatment,mid-treatment,post-treatment and clinical remissions or relapse throughout the entire treatment had been traced.At each stage the expression of methylated Id4 gene mRNA was detected by methylation-specific polymerase chain reaction (MS-PCR) and compared with the control group.The control group consisted of 20 non-neoplastic hematologic disorder affected children as sample donors.Results MS-PCR detection:in pre-treatment stage,there were 27 patients who were found methylated or partially methylated Id4 genes.Methylated ratio was thus at 64.3％ (27 patients out of a total of 42 patients).Those 27 patients were actively traced down along with different stages of treatment (21 NHL patients,6 HD patients).Before NHL there was 55.6％ methylated Id4 gene (15 cases out of 27 NHL patients).During chemotherapy treatment,there was 51.9％ of methylated Id4 gene positive (14 cases out of 27 patients).In post chemotherapy treatment,there was 48.1％ of methylated Id4 gene positive (13 cases out of 27 patients).Totally there were 9 patients showed clinical recovery after chemotherapy.There was 44.4％ of traceable methylated Id4 gene after recovered chemotherapy patients (4 recovered patients still carrying positive reading of methylated Id4 gene out of totally 9 recovered patients).There were 2 patients relapsed,with traceable methylated Id4 gene re-appeared in them afterwards.Throughout different treatment stages,there was no significant correlation in the treatment result and the appearance of methylated Id4 gene in early treatment stages (all P ＞ 0.05).But in latter treatment of chemotherapy,the correlation started to emerge (P ＜ 0.05) ; The overall statistics on NHL and HD share the same statistical pattern on different stages (all P ＞ 0.05).The control group had 20 patients,none of them had methylated Id4 gene.The study group showed noticeable difference in methylated Id4 gene before and after the experiment (P ＜ 0.001).RT-PCR result showed that before chemotherapy treatment,all of those who carried methylated Id4 gene had no expression of mRNA,by comparison to the control1 group which all had expression of mRNA.Conclusions Methylated Id4 gene is closely related in affected children who suffer from malignant lymphoma and its complications.The expression of Id4 gene is depressed when it has been methylated.The state of methylated Id4 gene is changed as patient's condition changed,so the methylated Id4 gene is thus a possible indicator of early diagnostic tool for children lymphoma.