Objective: To investigate the role of apoptosis-inducing factor (AIF) in TAp63γ-induced apoptosis. Methods: Plasmid pcDNA3. 1-TAp63γ was transfected into human esophageal squamous cancer EC9706 cells for 24 h. Apoptosis of EC9706 cells was detected by observing DNA fragmentation. The apoptotic rate was measured by flow cytometry. The translocation of AIF in EC9706 cells was studied with Mitochondrial/Cytosol/Nuclear extraction method. The plasmid that expressed hairpin RNA specific for AIF were constructed using chemosynthesis and gene recombination technology, and the interfering efficacy of AIF shRNA was detected by western blotting. After siRNA interference the apoptotic rate of EC9706 cells transfected with pcDNA3. 1-TAp63γ was analyzed by flow cytometry. Results: Agarose gel electrophoresis revealed that DNA ladder, a typical feature of apoptosis, was observed in the pcDNA3. 1-TAp63γ plasmid-transfected cells 24 later, but not in the pcDNA3. 1 plasmid-transfected cells. The apoptotic rate was 13.64% and 1. 37% after EC9707 cells were transfected with pcDNA3. 1-TAp63γ and pcDNA3. 1 plasmid, respectively. Signals of AIF-immunostaining were detected in the nucleus and cytosol proteins of pcDNA3. 1-TAp63γ transfection group, but not in pcDNA3. 1 transfection group. The plasmid expressing shRNA specific for AIF was successfully constructed and the interfering efficiency was about 80%. The apoptotic rate in AIF siRNA group was 4.52% at 24 h after transfection with pcDNA3. 1-TAp63γ. Conclusions: TAp63 γ gene expression induces the apoptosis of EC9706 cells. Down-regulation of AIF protein expression decreases the apoptosis induced by TAp63 γ.

Objective To evaluate the antitumor effect of corosolic acid and its impact on CAM and YSM angiogenesis.Methods The effects of CRA on A549 proliferation was studied by MTT method in vitro.Tumor-bruden nude mice model were established by injecting A549 lung cancer cell marked with bioluminescent to nude mice,and the growth of tumor in mice were detected by IVIS small animal in vivo imaging system.Experiment of chicken embryos eggs are used to observe the role of drugs on CAMand YSMblood vessels. Results The value of IC50 of CRA for A549 in vitro was 26.8μg/mL.A tumor-burdened animal experiment results showed that the CRA to A549 solid tumor has a certain therapeutic effect.CAM and YSM blood vessels of chicken embryos eggs treated with CRA were decreased significantly than negative control group.Conclusion CRA has certain therapeutic effect for A549,which may be related to the inhibition of angiogenesis in tumor tissues.

Objective To investigate the signal pathway of platelet activation in acute respiratory distress syndrome (ARDS). Methods Thirty healthy Sprague-Dawley (SD) rats were randomly divided into control group (n = 6) and model group (n = 24). The model of ARDS was reproduced by intravenous injection of oleic acid (0.25 mL/kg), and the rats in control group were injected with the same amount of normal saline. The blood of abdominal aorta was collected at 2, 6, 24, and 72 hours after model reproduction, the platelets were separated, and c-Jun N-terminal kinase phosphorylation (pJNK) levels which was one of major protein kinases in the mitogen-activated protein kinases (MAPKs) signal pathway was determined by Western Blot. The rats were sacrificed, the lung tissues were harvested, and lung coefficient (lung weight/body weight ×100%) and lung wet/dry (W/D) ratio were calculated. Pathological changes in the lung tissue were observed with hematoxylin-eosin (HE) staining in light microscope. Results Comp ared with the control group, platelet pJNK level in ARDS model group was significantly increased at 2 hours after model reproduction (gray value: 0.72±0.09 vs. 0.22±0.01), and peaked at 6 hours (gray value: 0.91±0.03 vs. 0.22±0.01), then it was decreased gradually. It was also significantly higher than that of control group till 72 hours after model reproduction (gray value: 0.39±0.06 vs. 0.22±0.01, all P < 0.05). Lung coefficient and lung W/D ratio in ARDS model group were significantly increased at 2 hours after model reproduction as compared with those of control group [(1.30±0.20)% vs. (0.60±0.10)%, 6.00±0.60 vs. 3.30±0.30], then they were decreased gradually. They were also significantly higher than those of control group till 72 hours after model reproduction [(0.90±0.10)% vs. (0.60±0.10)%, 4.80±0.70 vs. 3.30±0.30, all P < 0.05]. It was showed by light microscopy that lung tissue of rats in the control group had no significant pathological changes. At 2 hours after model reproduction in model group, clearly visible alveolar edema and interstitial edema, interstitial lung infiltration of inflammatory cells, small blood vessels dilation and congestion were found, and the re were a lot of protein exudates. The lesions of lung peaked at 24 hours. At 72 hours, absorption of most of fluid leaking in alveolar, alveolar space narrow, alveolar septum thickening, the reduction of inflammatory cells infiltration, fibrous tissue proliferation, and micro thrombosis formation were found. Conclusion In ARDS, in addition to pathological changes in the lung tissue, platelet activation occurs, and its activation process is related to the priming of JNK signal transduction pathways.

Background and purpose: Uveal melanoma (UM) is the most common primary intraocular malignancy in adult. Due to a high tendency for early metastasis the treatment of UM is very difficult. This study aimed to explore an effective approach for the treatment of patients with UM, we designed a strategy that combined HtrA2 gene therapy and radiation therapy. Methods:pIRES-Egr1-Omi/HtrA2 (pEgr1-HtrA2) recombinant plasmids were constructed and transfected into human UM cells (OCM-1) in vitro. The transfected cells were exposed to irradiation. HtrA2 mRNA and protein levels were detected by qRT-PCR and Western blot respectively. Assays that evaluated the apoptosis inducibility caused by HtrA2 gene therapy combined with radiation was performed by lfow cytometry. Followingly, the effects of HtrA2 overexpression on the in vitro radiosensitivity of uveal melanoma cells were investigated by clonogenic formation assay. The in vivo effects of HtrA2 gene therapy combined with radiation therapy were evaluated in different groups. Results:The recombinant plasmids could be successfully transferred into OCM-1 cells and transfection of pEgr1-HtrA2 plasmids combined with radiotherapy caused dramatically elevation of HtrA2 compared with non-irradiation cells in mRNA and protein levels, which was associated with increased apoptosis.Furthermore, we observed that the transfection of pEgr1-HtrA2 could significantly enhance radiosensitivity of OCM-1 cell in vitro. In mice bearing xenograft tumors, pEgr1-HtrA2 combined with radiation therapy signiifcantly inhibited tumor growth compared with the other treatment groups (P<0.05). Conclusion:Our ifndings indicate that radiation-inducible gene therapy may have potential to be a more effective and speciifc therapy for uveal melanoma because the therapeutic gene can be spatially or temporally controlled by exogenous radiation.

Background Postoperative endophthalmitis following intraocular lens(IOL)implantation is still one of the most feared complications of cataract surgery.Bacterial adhesion to IOLs during their insertion is a prominent etiological factor.The adhesion characteristics of bacteria to IOL are very vital for the prevention of endophthalmitis after cataract surgery.Objective The present study was to observe the in vitro adherence ability of Staphylococcus epidermidis to different intraocular lenses(IOLs)and compare the results in bacterial counting between scanning electron microscopy(SEM)photographs and quantitative cultures. Methods Five types of IOLs,including hydrophobic acrylic IOL,polymethylmethaerylate(PMMA)IOL,heparin-surface-modified(HSM) PMMA IOL,silicone(SI)IOL and hydrophilic acrylic IOL,were put into S.epidermidis(ATCC 12228)suspension for 1 hour.The bacterial adhesion numbers on the IOL surfaces were counted by quantitative cultures and scanning electron microscopy(SEM) photographs. Results Quantitative culture counting of viable adherent bacteria released by sonication showed that hydrophobic acrylic IOL and PMMA IOL were more likely for bacteria to attach.The number of bacteria on the five types of IOL surfaces showed significant differences(F=100.084,P=0.000).No significant differences were found in the number of bacteria between hydrophilic acrylic IOL and HSM-PMMA IOL (t=2.285,P=0.052)with the quantitative culture method.Direct counting of adherent bacteria in SEM photographs revealed that there were significant differences in bacterial adhesion numbers among difierent IOL material groups,with the numbers from high to low as follows:Hydrophobic IOL＞PMMA IOL＞SI IOL＞Hydrophilic IOL＞HSM-PMMA IOL(F=118.065,P=0.000).The counting method by SEM method was superior to that by quantitative cultures (t=5.019,P=0.000). Conclusion The bacterial adhesion ability varies upon the difference of IOL materials.Less bacterial adhesion is found on hydrophilic acrylic IOL and HSM-PMMA IOL,implying that the use of IOLs made from these two materials during surgery could diminish the incidence of postoperative endophthalmitis and intraocular inflammation associated with IOLs implantation.

Objective To evaluate the clinical application of sweep pattern visual evoked potential in assessing visual acuity (SPVEP-A)of amblyopic children.Design Prospective clinical study.Participants 80 patients(148 case)of amblyopic children and 26 children(52 case)with normal vision.Methods The correlation were analysed of SPVEP-A and E acuity in 80 amblyopic children and of SPVEP-A and PVEP-A in 26 children of normal visual.SPVEP-A was recorded with UTAS-E 3000(LKC)using gratings of 25 differ- ent spatial frequency from 0.2 to 18.8cpd as stimulus.The response were averaged and DFT on the monitor display.SPVEP-A was de- termined by extrapolate to 0 response amplitude.Both transient PVEP-A and SPVEP-A were examined in 26 normal children.The ob- jective visual acuity in PVEP were determined with the highest spatial frequency which evoked a recognizable response.Main Out- come Measures SPVEP-A,PVEP-A and E acuity.Results SPVEP-A were from 0.35 to 0.9 among 80 patients with corrected E acu- ity from 0.1 to 0.8.The correlation coefficient of SVEP-A-I with E acuity was 0.602 .The correlation coefficient of SPVEP-A.between reproducibility detection was 0.448.The correlation between SPVEP-A and E acuity in amblyopic slightly was higher than middle and severe amblyopia(r=0.773 vs r=0.590).In 26 normal children a correlation coefficient of 0.679 was obtained between E acuity anti PVEP-A but 0.424 between E acuity and SPVEP-A.Higher evaluation or lower evaluation of E acuity was found in SPVEP-A in this study.Acuity was upostimated in lower scope of E acuity and underestimated in higher scope of E acuity.The reproducibility and vari- ation were not satisified.Conclusion The sweep VEP is a objective method of amblyopic screening.There is significant positive corre- lation between SPVEP-A and E,but its variability is more great,so we should improve its stability and positive rate uheriorly.

Objective: To compare two methods (microbial assay and radioimmunoassay) for measuring plasma folate concentrations, and to examine the relationship between plasma folate levels, and alcohol consumption, tobacco use and body mass index, and the risk of hyperhomocysteinemia in China. Methods: We used a microtiter plate microbial assay and a radioimmunoassay to measure the folate concentration in 88 plasma samples. After comparing the results of these two methods and fitting a regression line, we examined the geographical, seasonal, and gender differences in folate concentration of plasma collected from 2 422 adults in south and north areas in China, and evaluated the association of plasma folate concentration, with alcohol consumption, cigarette smoking, and body mass index, and with the risk of hyperhomocysteinemia, using the data from the two assays. Results: The data from the two assays had a linear relationship ( r =0.879, P =0.000); the regression was Y =0.683 X +0.308 (where X and Y were nature logarithmic transformations of plasma folate by microbial assay and radioimmunoassay, respectively); however, the mean plasma folate levels by microbial assay were much higher than those obtained by radioimmunoassay. Both data sets showed similar plasma folate distributions among Chinese adults, associations with other risk factors, and the risk of hyperhomocysteinemia. We estimated that 19.9% of the Southerners and 67.1% of the Northerners had plasma folate concentrations by radioimmunoassay lower than the 6.8 nmol/L used to define plasma folate deficiency. Conclusion: There is a linear relationship between plasma folate levels determined by microbial assay and radioimmunoassay, but because of the different levels obtained in the two assays, it is difficult to use the microbial assay results to evaluate folate status at this time. The use of 10.5 nmol/L as a cut off for plasma folate deficiency by microbial assay needs further study.

Objective Carbon dots (CDs) are an emerging carbon nano-material which is environmentally-friendly, economical , efficient and stable .Their fluorescence properties can match the semiconductor quantum dot .Moreover , CDs have more excellent biocompatibilities .The purpose of this experiment is to apply CDs to the fluorescent immune probe to make them a new label , which can replace the traditional fluorescent dyes .Methods Using microwave heating method , the high strength fluorescent carbon dots were prepared .Wtih the EDC coupling method , the high strength fluorescent car-bon dots could bond with Escherichia coli antibodies to form a complex immune fluorescent probe .Specific recognition exper-iments were carried out in the model of E.coli O157∶H7.Results CDs were successfully applied to immune recognition of E.coli O157∶H7 and multicolor fluorescence was observed .Conclusion CDs can serve as a label of the fluorescent im-mune probe , and are expected to become a new type of low toxicity biosensor with independent intellectual property rights .

Colonoscopy has been accepted as the standard method for evaluation of colon and rectum,its success rate depends on the quality of bowel preparation. Aims:To evaluate the cleanliness and tolerance of fractionated dose versus single dose polyethylene glycol electrolyte solution( PEG-ES) bowel preparation regimens for colonoscopy. Methods:A total of 427 consecutive asymptomatic individuals undergoing colorectal cancer screening were enrolled and randomly assigned into 2 groups. Subjects in group A drank 1. 5 L PEG-ES on the eve and 4 hours before colonoscopy, respectively;subjects in group B received a single dose of 3 L PEG-ES 5 hours before colonoscopy. Score and degree of Boston bowel preparation scale(BBPS)and PEG-ES related adverse effects of the two groups were assessed and compared. Results:There were no significant differences in gender,age and cecal insertion rate between group A and group B(P ﹥ 0. 05). Score of BBPS was significantly higher in group A than in group B(P ﹤0. 01). Both regimens met the requirement of conventional colonoscopy,however,the cleanliness of colon was graded as excellent in more subjects of group A( P ﹤ 0. 01),and less subjects of group A complained PEG-ES related nausea(P ﹤0. 05). Logistic regression analysis revealed that the PEG-ES drinking pattern was associated with cleanliness of colon and occurrence of nausea( P ﹤ 0. 05). Conclusions:Fractionated dose PEG-ES regimen provides a better colonic cleansing quality and tolerance for bowel preparation of colonoscopy,which is superior to that of single dose regimen.

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.