Objective To study the inhibitory effects of pigment epithelium derived factor (PEDF) on oxygen-induced retinal neovascularization in mice,and to investigate the possible involvement of interleukin-1β (IL-1β) in the neovascular-inhibitory function of PEDF.Methods A total of 140 postnatal day (P)7 C57BL/6 mice were randomly divided into normal control group,oxygen-induced retinopathy (OIR) model group,PEDF treatment group and PBS treatment control group.All mice except normal control group with their mothers were exposed to (75 ± 2)％ oxygen environment for 5 days and then kept in room air for another 5 days to establish the OIR model.Mice in normal control group were kept in room air only.At P12 and P14,respectively,mice in PEDF treatment group received intravitreous injections of 1 μl PEDF (2 μg/μl),while PBS treatment control group received the same volume of PBS (10 mmol/L,pH7.4).All mice were euthanized at P17 and eyes were isolated.The changes of retinal vessels were observed on retinal flat mounts and cryosections by fluorescence microscopy.Retinal specimens were prepared for IL-1β protein and mRNA analysis by Western blot and real time fluorescence quantitative reverse transcription-polymerase chain reaction (Real-time RT-PCR).Results Changes of retinal vessels had been viewed by fluorescence microscopy on flat-mounted retina,the relative retinal neovascularization areas were significantly increased in OIR model group compared with normal control group (t =15.02,P＜0.01),and the relative retinal neovascularization areas were obviously smaller in PEDF treatment group than those in PBS treatment control group (t=5.96,P＜0.01).Fluorescence staining revealed that retinal vascular tufts were extending from outer plexiform layer (OPL) to ganglion cell layer (GCL) of the retina along with multiple interconnections; Neovascular tufts in OIR model group and PBS treatment control group were presenting distinctly more than those of normal control group and PEDF treatment group.The specific expression levels of IL-1β protein in retinas of OIR mice by Western blot analysis were higher than those of normal control group(t=3.35,P＜0.05),While these of PEDF treatment group showed a considerable decline in comparison with PBS treatment control group (P＜0.01),and there were no difference in normal control group and PEDF-treated group (F=11.764,P＞0.05).Similarly,expression levels of IL-1β mRNA tested by Real-time RT-PCR were obviously increased in the OIR model group when compared to normal control group(t =4.43,P ＜ 0.01).After treated with PEDF,expression levels of IL-1β mRNA showed a considerable decrease when compared to PBS treatment control group (P ＜ 0.01),and there were no difference in normal control group and PEDF-treated group (F=11.15,P＞0.05).Conclusions PEDF can inhibit oxygen-induced retinal neovascularization.The mechanism may be related to that PEDF can downregulate the expression of IL 1β in retina.

Objective To discuss the electrocardiogram(EKG)positioning method to the guiding role of determining the pipe length and the accuracy of operative localization in central venous catheterization procedure. Methods Chose 32 cases of tumor patients who had center venipunture.Use the catheter taken EKG data when cath-etering,and then given validation using postoperative chest X -ray or fluoroscopy.Judgment the sensitivity,specificity and disposable catheters success rate of the EKG positioning method.Results In the 32 cases of cancer patients, 30 patients had characteristic P waves,when the chest X -ray confirmed the superior vena cava or the junction with the right atrium,one case into the right atrium,when one case of non -P -wave in the subclavian after intravenous discounts tune into the tube after it confirmed the superior vena cava.Conclusion EKG positioning method with high accuracy in the deep venous catheter in the catheter tip positioning applications.The clinical applications are feasible.

Objective To investigate the effect of active silver ions liquids on incontinence of patients with cerebrovascular diseases.Methods Forty-eight patients with incontinence from cerebrovascular diseases were randomly divided into the treatment group and control one in equal number:the control group was managed with washing and cleaning the perineum with warm water in the morning and evening and the treatment group with active antibacterial liquid silver ion liquids in the afternoon once a day.The two groups were compared in terms of red swelling,itching,papula,blister,local shallow ulcer and abscess in the skin around the perineum as well as the incidence of urinary system infections.Results The treatment group had significantly lower incidences of red swelling,itching,papula,blister,local shallow ulcer and abscess in the skin around the perineum as well as the incidence of urinary system infections,as compared to the control group(all P<0.05).Conclusion Cleaning perineum with active silver ions liquids is effective for the prevention of infections in the skin around the perineum and urinary system infections.

Objective To explore the effects of SLC concentration gradient on suppression of tumor immune escape. Methods According to different SLC concentration, there were six groups. The HLA-Ⅰexpression and apoptosis of MCF-7 were detected with FCM,and intracellular BCL-2 expression was analyzed by western blot. The production of TGF-? was detected with ELISA. Results In a certain range of concentration gradient, following SLC increase, HLA-Ⅰexpression level on MCF-7 was improved, and apoptosis was induced but BCL-2 expression was enhanced. Moreover, the secretion of TGF-? was suppressed. Conclusion SLC inhibites tumor immune escape.

Objective: To observe the effect of a novel targeted agent enzastaurin alone or in combination with gefitinib on ge-fitinib-resistant human non-small cell lung cancer cells to explore the rational drug combination. Methods: CCK-8 assay was used to measure the cell proliferation. Combination index ( CI) was calculated by Chou-Talalay method to assess the efficacy of the combination therapy. The flow cytometry (FCM) was used to analyze the change in the cell cycle. Results:In 72h, the IC50 value of gefitinib and enzastaurin for the lung cancer NCI-H460 cells was 6. 99μmol·L-1 (95%CI:3. 55-13. 79μmol·L-1 ) and 7. 25μmol·L-1 (95%CI:4. 77-1. 02 μmol·L-1), respectively. The inhibition effect on the cell proliferation was stronger in the combination treatment than that in the monotherapy (P<0. 05), and the simultaneous treatment showed the most significant inhibition effect (P<0. 01). The IC50 value of gefitinib for H460 cells in the simultaneous administration group, the sequential administration group with gefitinib used first and the sequential administration group with enzastaurin used first was 0.006 μmol·L-1(95%CI:0.002-0.020μmol·L-1), 0.02μmol·L-1(95%CI:0.011-0.037 μmol·L-1) and 0.085 μmol·L-1(95% CI:0.042-0.170μmol·L-1, respectively. The CI of the simultaneous administration group was lower than one when the gefitinib concentration was above 0. 05μmol·L-1 . The cell cycle distribution result indicated that the simultaneous administration group had significantly increased G0/G1 proportion (P<0. 05) and induced cell cycle arrest at G1 phase. Conclusion:Protein kinase C inhibitor enzastaurin combined with EGFR inhibitor gefitinib shows a synergistic effect, suggesting that the combination treatment of the two drugs might be a new strategy for the follow-up therapy of gefitinib-resistant non-small cell lung cancer.

A gram-negative bacillus was isolated from the bodies of dead clams among clambanks along the south beach of Jiangsu province. It was confirmed to the Vibrio furnissii by morphological and biochemical characteristic examinations. This isolate grows well in clam's body fluid media and may multiply rapidly in sea-water at temperature about 25—37℃. It is of high toxicity. The healthy clams were all diseased with the same syndrome and died as natural illness by inoculate them with the isolate. It is considered that the large enormous death of clams along the south beach of Jiansu province is concerned with the wide spreading of the clam's infectious disease caused by Vibrio furnissii which hasn't been reported both internal and abroad.

Background Bevacizumab and triamcinolone acetonide (TA) has been widely used in the treatment of diabetic macular edema (DME) clinically,but the effectiveness of both treatment has disadvantage.Therefore,some researchers try to combine bevacizumab with TA for the management of DME,but its efficacy is controversial.Objective This study was to evaluate the efficacy and safety of intraovitreal injection of bevacizumab combined with TA versus bevacizumab for DME.Methods The randomized controlled trials (RCTs) of bevacizumab combined with TA versus bevacizumab via intraovitreal injection for DME were searched from Pubmed,EMbase,Cochrane Library,CNKI.The methodological quality of the literature was evaluated according to evidencebased medicine (EBM),and the quality of the RCTs was appraised based on the Cochrane Handbook for Systematic Reviews of Interventions Version 5.0.The outcome indicators including the change values of central macular thickness (CMT) and best-corrected visual acuity (BCVA) as well as the safety indicators including topical and system adverse response of RCTs were analyzed with Cochrane Collaboration' s software RevMan 5.0.Results Nine RCTs were included with 665 eyes.The decrease value of CMT was more remarkable in the bevacizumab combined with TA group than that of the only bevacizumab group 12 weeks and 18 weeks after intravitreal injection (WMD =-44.69,95％ CI:25.27-64.11,P ＜ 0.000 001 ; WMD =-66.86,95％ CI:40.67-93.05,P ＜ 0.000 001).However,no significant differences were found in the change value of CMT in 6 weeks and 6 months after injection between the two groups (WMD =-15.40,95％ CI:-4.04-34.85,P =0.12 ; WMD =-2.57,95％ CI:-19.62-24.75,P =0.82).The improvement value of BCVA (LogMAR) in the bevacizumab combined with TA group was superior to that of the only bevacizumab group 6 weeks after injection (WMD =-0.04,95 ％ CI:-0.08--0.00,P =0.05),but there were no significant differences between the two groups at 12weeks,18 weeks and 6 months after treatment (WMD =-0.04,95％ CI:-0.12-0.05,P=0.36;WMD =-0.04,95％ CI:-0.11-0.03,P=0.28; WMD =0.03,95％ CI:-0.05-0.12,P=0.45).The incidence rate of transient anterior response after injection was not significantly different between the two groups (RR =0.89,95％ CI:0.49-1.60,P =0.70).Secondary ocular hypertension after injection occurred in 30 eyes in the bevacizumab combined with TA group,but no hypertension was seen in the only bevacizumab group.Conclusions Compared with only bevacizumab,intravitreal injection of bevacizumab combined with TA has a better efficacy in improving CMT but no obvious dominant in increasing BCVA for early DME.Intravitreal injection of bevacizumab combined with TA seemingly has a higher risk of inducing controllable ocular hypertension than administration of only bevacizumab.

Aim To investigate the protective effects of MANF on human neuroblastoma SH-SY5 Y cells suf-fering from oxygen-glucose deprivation/reperfusion ( OGD/R) and the underlying mechanism. Methods SH-SY5Y cells were treated with OGD for 6 h, fol-lowed by reperfusion for 12 h. Meanwhile, the cells were incubated with 2 μmol · L-1 recombinant human protein MANF for 12 h during reperfusion. The cell morphology was observed under an optical microscope. The cell viability was determined by MTT assay. PI
staining was performed to detect the number of dead cells. Western blot was performed to determine the protein levels of endogenous MANF, glucose-related protein 78 ( GRP78/BiP) , phosphorylated inositol re-quiring enzyme 1 ( p-IRE1 ) , phosphorylated eukaryot-ic translation initiator factor 2α ( p-eIF2α) , cleaved caspase-3, and C/EBP-homologous protein (CHOP). Results The cells exposed to OGD/R became smaller and round, and the neurites of the cells were shortened or disappeared . Recombinant human protein MANF
improved the survival rate ( P <0. 05 ) and decreased the death rate ( P <0. 05 ) of SH-SY5 Y cells treated with by OGD/R. Western blot assay showed that the endoplasmic reticulum ( ER) stress-associated proteins GRP78/BiP, p-IRE1, p-eIF2α, and MANF were in-creased significantly after OGD/R treatment, compared with the untreated controls. However, the increases of secretion levels of apoptosis-associated proteins CHOP
and cleaved caspase-3 in SH-SY5 Y cells induced by OGD/R were significantly suppressed by MANF. Con-clusion OGD/R up-regulates the ER stress-associated proteins and causes apoptosis. MANF inhibits OGD/R-induced cell death, which may be related to attenua-ting ER stress-induced apoptosis.

Objective To examine the complement component 1 Q subcomponent-binding protein (C1QBP) gene expression in human resistance choriocarcinoma cell lines and its parental cell line JeG-3,and to investigate whether silence C 1QBP by small interference RNA could reverse the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.Methods Expression of C1QBP mRNA and protein in cells were detected by real-time fluorogenic quantitative PCR and western blot,respectively.The difference of C 1QBP expression was compared between human resistance choriocarcinoma cell lines and its parental cell line JeG-3.Sub-cellular location was proved by confocal immunofluorescence microscopy.A lentiviral vector containing short hairpin RNA (shRNA) targeting C 1QBP was constructed and cotransfected with the packaging plasmid mixture into 293T cells by lipofectamine 2000.The human resistance choriocarcinoma cell lines were infected with the packaged lentivirus.Real-time fluorogenic quantitative PCR and western blot were used to validate whether the C 1QBP gene expression was silenced.The cell counting kit 8(CCK8)was used to determine the drug sensitivity.Results (1)The C1QBP mRNA expression levels among four human resistance choriocarcinoma cell lines[JeG-3/floxuridiuum (FUDR),JeG-3/methotrexate (MTX),JeG-3/etoposide (VP),JeG-3/dactinomycin (KSM)] were 2.520±0.680,1.770±0.230,1.940±0.090 and 1.740±0.350 folds compared to that in JeG-3 cells.The C1QBP protein was higher expression level in human resistance choriocarcinoma cell lines than that in JeG-3.The immunofluorescence methods and confocal analysis showed that C1QBP localized predominantly in the mitochondrial matrix.(2)The C1QBP mRNA expression in JeG-3/FUDR cells after infected with lentiviral vector were decreased by 93.1％ (P＜0.01).The protein expression of C 1QBP in JeG-3/FUDR cells after infected with lentiviral vector were almost completely suppressed.The resistance indexes of four human resistance choriocarcinoma cell lines(JeG-3/FUDR,JeG-3/MTX,JeG-3/VP,JeG-3/KSM) were respectively 86.3％,93.9％,92.8％ and 89.9％,which were decreased remarkably by knockdown the C 1QBP expression (P＜0.05).Conclusions C1QBP is overexpressed in human resistance choriocarcinoma cell lines compared with parental cell line JeG-3.Inhibition of C 1QBP by lentivirus-mediated small interference RNA could effectively reverses the resistance of human resistance choriocarcinoma cell lines to its relevant chemotherapy drugs.

Objective To establish the platelet antigen panel for identifying the specificity of platelet antibodies which cause platelet transfusion refractoriness and neonatal alloimmune thrombocytopenia and provide evidence for clinical therapy and platelet genotyping research.Methods Based on the frequency distribution of human platelet alloantigen (HPA)-1 to HPA-16 gene in China, the frequencies of HPA-1 to HPA-6,HPA-15 alleles in blood group O donors were genotyped by the polymerase chain reaction with sequence-specific primers (PCR-SSP) method, and suitable donors were chosen to establish platelet-specific antigen panel.Using the established platelet-specific antigen panel, the specificity of platelet antibodies caused by alloimmune reaction was identified by using simplified sensitized erythrocyte platelet serology assay (SEPSA).Results Eleven ptatelet donors with blood group O were chosen to establish platelet-specific antigen panel which can identify specificity of HPA-1 to HPA-6, HPA-15 antibodies.One case of HPA-4b (Penb) and two cases of HPA-15a (Govb) platelet specific antibodies were detected in 1 120 samples.Conclusion Identifying the specific platelet antibodies using platelet specific antigen panel has profound significance on increasing the safety and effectiveness of clinical platelet transfusion and prevention of neonatal alloimmune thrombocytopenia.