Background The underlying mechanism of choroidal neovascularization(CNV) is multifactorial and complex.Focal adhesion kinase(FAK) plays a crucial role in controlling essential cellular processes and influencing distinct steps of the angiogenic response.But to our knowledge,seldom study on the effect of FAK on CNV formation has been reported previously.Objective In this study,the effect of several CNV risk factors on the expression of FAK in cultured retinal pigment epithelium(RPE) cells was investigated to illuminate effect of FAK on CNV.Methods Human RPE cells were isolated from donor eyes and exposed to H2O2,swallow of outer segment of photoreceptors(POS) and extracellular matrix(ECM) separately with the treating as follows:RPE cells were co-cultured with 10,20,50 and 100μmol/L H2O2 for 20 days;POS(1×106/ml) were co-cultivated with RPE cells for 20 days(setting control group,POS group,hypoxia group with 200μmol/L CoCl2,and POS+hyoxia group);RPE cells were cultured on the plates coated with 100mg/L fibronectin(FN),laminin(LN) or collagen typeⅠfor 30minutes or 1 hour.The expression of FAK and pFAK in RPE cells were examined by Western blot analysis.Results FAK was highly expressed in the 20μmol/L and 50μmol/L H2O2 groups compared with control group(P＜0.01);while he expression level of pFAK was reduced after treated with H2O2 in comparison with the control group(P＜0.01).After cultured with POS for 20 days,the undigested lysosome could be observed in RPE cells.The expressions of FAK and pFAK in RPE cells were not significantly changed between control group and POS groups(P＞0.05),but those in hypoxia group were significantly up-regulated in comparison with control group(P＜0.01).Compared with the hypoxia group,the expression amount of pFAK was elevated in POS+hyoxia group(P＜0.01).In comparison with control group,the increased pFAK expression was seen in FN,LN and collagen typeⅠtreating for 1-hour groups(P＜0.05,P＜0.01).Conclusion FAK pathway participates in several CNV-initiated signaling,such as H2O2,POS and ECM,in cultured RPE cells.It is reasonable to believe that FAK potentially plays an important role in CNV-dependent disorder.

OBJECTIVETo investigate the influence of pyrrolidine dithiocarbamate (PDTC) on the expression of transforming growth factor beta(1) (TGF-beta(1)), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor-1 of metalloproteinase (TIMP-1) in rats with pulmonary damage induced by paraquat (PQ).

METHODSFifty-four healthy male SD rats were randomly assigned into the control group (normal saline), the PQ-treatment groups (4 groups) and the PDTC treatment groups (4 groups). Except the rats in the control group, the rats in the PQ group were gavaged only with 40 mg/kg PQ, and PDTC group with 40 mg/kg PQ plus immediate injection 120 mg/kg PDTC (i.p). On the 3rd, the 7th, the 14th and 28th day after treatments, one group rats of each treatments were sacrificed and lung and blood samples were collected. The level of TGF-beta(1) protein in the plasma, the mRNA expression of TGF-beta(1), MMP-2 and TIMP-1 were evaluated using RT-PCR and real-time quantitative PCR, while pathological changes of lung were examined under optical microscope and electrical microscope.

RESULTSThe TGF-beta(1) protein, TGF-beta(1) and MMP-2 mRNA expression were increased significantly in the earlier stage and then decreased after PQ administration (P < 0.05 or P < 0.01), while the mRNA level of TIMP-1 was augmented continuously (P < 0.01) throughout the study compared to the control group. In comparison with the PQ group, in the PDTC treatment group, the TGF-beta(1) mRNA expression on the 3rd and the 14th day, 0.54 +/- 0.08 and 0.72 +/- 0.04 respectively, the MMP-2 mRNA expression on the 7th and 14th day, 1.62 +/- 0.50 and 1.97 +/- 0.34 respective-ly, and the TIMP-1 mRNA on the 7th and 21st day, 1.79 +/- 0.21 and 2.00 +/- 0.34 respectively, were significantly decreased (P < 0.05 or P < 0.01).

CONCLUSIONPDTC could attenuate paraquat-induced up-regulation of TGF-beta(1) and its mRNA expression, MMP-2 and TIMP-1 mRNA levels, which indicates that PDTC may exert its protective effects on paraquat-induced pulmonary damage by alleviating the earlier inflammation damage and adjust-ing the balance between MMPs and TIMPs. However, further studies are still warranted to investigate and clarify the underlying mechanisms involved in this complicated process.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Disease Models, Animal ; Lung ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Paraquat ; poisoning ; Pyrrolidines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates ; pharmacology ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; metabolism

OBJECTIVETo investigate the effect of handportable mobiletelephone microwave radiation on rat central nervous system by setting up rat model.

METHODS80 healthy male SD rats (weighed about 200 g) were divided into 4 groups at random: control, radiation, decranium, decranium + radiation. TUNEL method was adopted used to detect the apoptosis of neurons after irradiation, then immunohistochemistry was used to detect Bcl-2, Bax expression in all brain tissue.

RESULTSTUNEL positive rate, Bax and Bcl-2 positive cell numbers could be found in decranium + radiation group [(26.45 +/- 9.27)%, (23.5 +/- 3.58), (11.1 +/- 2.55) respectively]. There were significant differences among control [(9.59 +/- 2.55)%, 14.2 +/- 2.46, 7.0 +/- 1.14 respectively], decranium group [(9.52 +/- 1.93)%, 15.5 +/- 1.77, 7.4 +/- 1.76], radiation group [(10.04 +/- 3.62)%, 15.9 +/- 2.02, 7.2 +/- 1.07] (P < 0.01). But the difference was not found in the ratio of Bax to Bcl-2.

CONCLUSIONMicrowave radiation did not affect the intact rat, but did promote the occurrence of neuron apoptosis in cranial defect rat. Bax, Bcl-2 gene participated in regulation of apoptosis. The intact cranium may be an important factor to protect the neurons against handportable mobiletelephone microwave radiation to some extent.

Animals ; Apoptosis ; radiation effects ; Brain ; pathology ; radiation effects ; In Situ Nick-End Labeling ; Male ; Microwaves ; adverse effects ; Neurons ; pathology ; radiation effects ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein

Global longitudinal strain (GLS) at rest on two-dimensional speckle tracking echocardiography (2D STE) was demonstrated to help detect coronary artery disease (CAD).However,the optimal cut-off point of GLS and its diagnostic power for detecting critical CAD in non-diabetes mellitus (DM) patients are unknown.In the present study,211 patients with suspected CAD were prospectively included,with DM patients excluded.All patients underwent echocardiography and subsequently coronary angiography within 3 days.Left ventricular (LV) GLSs were quantified by 2D STE.Territorial peak systolic longitudinal strains (TLSs) were calculated based on the perfusion territories of the 3-epicardial coronary arteries in a 17-segment LV model.Critical CAD was defined as an area stenosis ≥70％ in ≥1 epicardial coronary artery (≥50％ in left main coronary artery).Totally 145 patients were diagnosed as having critical CAD by coronary angiography.Significant differences were observed in all strain parameters between patients with and without critical CAD.The area under the receiver operating charcteristic (ROC) curve (AUC) for GLS in the detection of left main (LM) or threevessel CAD was 0.875 at a cut-off value of-19.05％ with sensitivity of 78.1％ and specificity of 72.7％,which increased to 0.926 after exclusion of apical segments (cut-off value-18.66％;sensitivity 84.4％ and specificity 81.8％).The values of TLSs were significantly lower in regions supplied by stenotic arteries than in those by non-stenotic arteries.The AUC for the TLSs to identify critical stenosis of left circumflex (LCX) artery,left anterior descending (LAD) artery and right coronary artery (RCA),in order of diagnostic accuracy,was 0.818 for LCX,0.764 for LAD and 0.723 for RCA,respectively.In conclusion,in non-DM patients with suspected CAD,GLS assessed by 2D STE is an excellent predictor for LM or three-vessel CAD with high diagnostic accuracy,and a higher cut-off point than reported before should be used.Excluding apical segments in the calculation of GLS can further improve the predictive accuracy of GLS.It is unsatisfactory for TLSs to be used to identify stenotic coronary arteries.

Objective To observe the serum level of cyclopropaneoctanoic acid 2-hexyl (CPOA2H) in patients with hypertrilyceridemia-related disorders,and investigate its clinical significance.Methods 53 obese patients (Obese group),62 obese patients after a 3 month low calorie diet (Obese patients after low calorie diet group),46 patients with chronic kidney disease (CKD group),and 60 healthy controls (Control group) were collected from Mar 2013 to Dec 2015 in Shaanxi Provincial People's Hospital.Gas chromatography-mass spectrometry (GC-MS) were used to detect the serum fatty acid and CPOA2H level in four groups.Results Ages,body mass index (BMI),C-reaction protein (CRP),total cholesterol (TC),triacylglycerols (TAG) among four groups had significant difference (F=6.85 ～ 25.36,P＜0.05).Most saturated fatty acid (14 ∶ 0,16 ∶ 0) and monounsaturated fatty acid (14 ∶ 1,16 ∶ 1,18 ∶ 1,20 ∶ 1) in obese group were higher than controls (F=3.251～7.351,P＜0.05).The polyunsaturated fatty acid (18 ∶ 2n-6,20 ∶ 4n-6) in CKD group were lower than controls (F=4.351 ～6.251,P＜0.05).There existed significant difference of serum levels of CPOA2H among four group (F=19.95,P=0.005).Serum level of CPOA2H in control (r=0.63,P=0.033) and CKD group (r=0.61,P=0.044) were positively related with TAG.Serum level of CPOA2H in obese group and obese patients after low calorie diet group were positively related with TC (r=0.70,P=0.011;r=0.48,P=0.021) and TAG (r=0.75,P=0.024;r=0.72,P=0.018).Conclusion Hypertrilyceridemia-related disorders,such as obesity and CKD,presented with elevated serum levels of CPOA2H,it suggested that serum level of CPOA2H is positively related to serum TAG concentration rather than BMI.

Not Abstract.

OBJECTIVETo investigate the spatial learning and exploration along with the CNS excitatory amino acid neurotransmitters profiles in adult rats subchronically exposed to the anticholinesterase organophosphorus insecticide dimethoate.

METHODSRats were gavaged daily with dimethoate (0, 5, 10 or 20 mg/kg via oral) in NS. for 90 days. Morris water maze tasks were used to test the spatial learning and memory in the rats after the dimethoate exposure. Simultaneously, rats were decapitated for the determination of brain cholinesterase AChE activities, glutamate concentrations, and the NMDA receptor NMDA-R densities and affinities in hippocampus.

RESULTSLatencies to find a hidden escape platform were significantly longer in dimethoate dosed groups than that of the control group in the place navigation tests. Subsequently, the times of crossing the location of platform which had been removed obviously decreased in the highest dose group compared with that of the control in the spatial probe tests (P < 0.05). AChE activity was significantly reduced 42% approximately 78% by all three doses of dimethoate (P < 0.05). Glutamate concentrations were increased significantly 132.9% approximately 134.5% by the two highest doses of dimethoate (P < 0.05). In addition, the NMDA receptor bindings were reduced 21.2% approximately 23.2% with the statistical significance at the same two highest doses (P < 0.05). Furthermore, the receptor affinities was reduced 33.1% by the highest dose group (P < 0.05). The lesions of spatial memory were statistically corrected with the decrease of the NMDA-R affinities (P < 0.05).

CONCLUSIONThe cholinergic lesion as well as the excitatory amino acid system alteration might attribute to the inferior ability in spatial learning and memory in dimethoate subchronically exposed rats.

Acetylcholinesterase ; metabolism ; Animals ; Chronic Disease ; Dimethoate ; toxicity ; Disease Models, Animal ; Glutamic Acid ; metabolism ; Insecticides ; toxicity ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Toxicity Tests, Subchronic

OBJECTIVETo investigate whether the p38 mitogen-activated protein kinase (MAPK) signaling pathway mediates advanced oxidation protein products (AOPPs)-induced epithelial-to-mesenchymal transition (EMT) in tubular cells.

METHODSHuman proximal tubular cells (HK-2 cells) exposed to AOPP-bovine serum albumin (BSA) were examined for expressions of p38 MAPK and phosphorylated p38 MAPK using Western blotting. Western blotting and quantitative RT-PCR were used to examine the protein and mRNA expressions of EMT markers E-cadherin and vimentin and endoplasmic reticulum stress marker glucose-regulated protein (GRP) 78 in cells treated with SB203580 (an inhibitor of the p38 MAPK signaling pathway) prior to AOPP exposure. The cells treated with AOPPs following pretreatment with salubrinal (an inhibitor of endoplasmic reticulum stress) were also examined for expressions of p38 MAPK and phosphorylated p38 MAPK.

RESULTSAOPP treatment induced the phosphorylation of p38 MAPK in HK-2 cells. AOPP-induced decrease in E-cadherin expression and overexpression of vimentin and GRP78 were partly inhibited by pretreatment of the cells with SB203580. Salubrina partly suppressed AOPP-induced phosphorylation of p38 MAPK in the cells.

CONCLUSIONp38 MAPK signaling pathway, which is regulated by endoplasmic reticulum stress, might mediate AOPP-induced EMT in HK-2 cells.

To investigate the effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor simvastatin (SV) on proliferation, apoptosis and the PI3K/AKT signaling pathway in human acute monocytic leukemia cell line SHI-1. SHI-1 cells were incubated with different concentrations of SV (5, 10, 15 µmol/L). Otherwise, SHI-1 cells without any treatment were used as control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detection. MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the early stage apoptosis ratio. The human PI3K-AKT Signaling Pathway RT(2) Profiler(TM) PCR Array was used to detect the expression of 84 genes involved in PI3K-AKT signaling. The results indicated that the SV inhibited the proliferation and inducted the apoptosis of SHI-1 cells in time- and dose-dependent manners significantly. The growth inhibition rates of SHI-1 cells treated with 15 µmol/L SV for 24, 48 and 72 h were 26.82, 47.09 and 63.92, respectively; and their early stage apoptosis ratios were 5.75, 13.25 and 15.59, respectively. Compared with the control group, expression levels of 39 genes were changed in the group of 15 µmol/L SV at 48 h, among them 26 genes were down-regulated and 13 genes were up-regulated. It is concluded that the SV can inhibit proliferation and induce apoptosis of SHI-1 cells, and the mechanism may be associated with the changes of gene expression level in PI3K-AKT signaling pathway regulated by SV.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Monocytic, Acute ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; Simvastatin ; pharmacology

OBJECTIVETo investigate the effect and mechanism of arsenic trioxide on hepatoma cell line BEL-7402 and observe the effect and best administration method of arsenic trioxide on hepatocellular carcinoma patients who were not suitable for operation.

METHODSThe cell activity and morphologic changes were studied after being treated with arsenic trioxide in different concentrations. The apoptosis was detected by flow cytometry assay and DNA fragmentation assay. The caspase-3 level of mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and fluoro-spectrophotometer. The growth inhibition of implant tumor was observed in nude mice treated with arsenic trioxide in different concentrations. Arsenic trioxide was used in hepatocellular carcinoma patients by intravenous dropping and continuous regional infusion through hepatic artery.

RESULTSThe effect of arsenic trioxide on hepatoma cell lines was dependent on the time and concentration obviously. The decrease in cell number was preceded by morphological changes in the treated BEL-7402 cells that were characteristic of apoptosis, including membrane blebbing, shrunken cytoplasm, nuclear condensation and loss of adhesion. Flow cytometry assay showed an arrestment at G(2)/M phase and sub-G(1) cell peak. DNA fragmentation assay showed a marked DNA ladder. The mRNA level of caspase-3 was no change in RT-PCR whereas the protein of caspase-3 was increased after added As(2)O(3) 1 - 36 h. Caspase-3 activity began to increase after 2 h and reached a maximal level after 12 h in a linear fashion. Then, the level of caspase-3 was decreased, but still in a high level. The growth inhibition of implant tumors was obviously in nude mice. The intravenous usage of arsenic trioxide could improve the quality of life with low toxicity in hepatocellular carcinoma patients not suitable for operation. The tumor size decreased in 30 patients and AFP value decreased in 19 patients by continuous regional infusion through hepatic artery.

CONCLUSIONSArsenic trioxide can obviously inhibit the growth of hepatoma cell line BEL-7402 through inducing hepatoma cell apoptosis. The activation and increase of Caspase-3 is the possible mechanism of apoptosis, and the acting point is in pro-enzyme level. The best result of arsenic trioxide on non-operative patients should be gotten in continuous infusion through hepatic artery.

Adult ; Aged ; Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; therapeutic use ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Line, Tumor ; Chemotherapy, Cancer, Regional Perfusion ; Female ; Humans ; Liver Neoplasms ; drug therapy ; pathology ; Liver Neoplasms, Experimental ; pathology ; Male ; Mice ; Mice, Nude ; Middle Aged ; Oxides ; pharmacology ; therapeutic use