AIM:To study the protective effect of anti-aging Klotho protein on human umbilical vein endothe-lial cells (HUVECs) treated with high glucose (HG).METHODS:HUVECs were cultured in vitro, and divided into PBS control group, 5.5 mmol/L glucose group, 33.3 mmol/L glucose group, 0.1 μmol/L Klotho +33.3 mmol/L glucose group, 1 μmol/L Klotho+33.3 mmol/L glucose group , and 10μmol/L Klotho+33.3 mmol/L glucose group .The viabili-ty of the HUVECs was measured by MTT assay .The content of malondialdehyde ( MDA) , and the activities of lactate de-hydrogenase (LDH), superoxide dismutase (SOD) and glutathione (GSH) in cell culture supernatants were observed . The production of reactive oxygen species ( ROS) in HUVECs was analyzed by flow cytometry .The levels of nitric oxide ( NO) , endothelin ( ET-1 ) , intercellular adhesion molecule-1 ( ICAM-1 ) in HUVEC culture medium were detected by ELISA.The protein expression of nuclear factor-kappa B (NF-κB) in the HUVECs was determined by Western blot .RE-SULTS:Compared with PBS control group , 33.3 mmol/L glucose significantly decreased the HUVEC viability , increased ROS, LDH and MDA levels , reduced the activities of SOD and GSH , decreased the NO secretion , and induced the ET-1 and ICAM-1 secretion and the protein expression of NF-κB in HUVECs.When HUVECs were treated with Klotho protein at different concentrations combined with 33.3 mmol/L glucose, the cell viability was increased significantly , the ROS, LDH and MDA levels were decreased significantly , the antioxidant SOD and GSH activities were significantly increased , the se-cretion of NO was increased , but ET-1 and ICAM-1 releases and protein expression of NF-κB were significantly reduced . CONCLUSION:Anti-aging Klotho protein promotes the viability of HUVECs treated with HG , reduces the oxidative dam-age and ROS production , and restores the normal secretory function of HUVECs , thus playing a protective role in vascular endothelial cells through reducing the protein expression of NF-κB.

Objective To investigate the incidence and relevant factors of upper gastrointestinal bleeding(UGB)in patients with coronary heart disease(CHD)undergoing dual antiplatelet therapy(DAT)with aspirin plus clopidogrel.Methods A total of 391 CHD patients treated with aspirin plus clopidogrel (DAT group)and 502 control CHD patients receiving single antiplatelet therapy(SAT)with aspirin or clopidogrel(SAT group)were enrolled.The incidence and relevant factors associated with UGB were retrospectively analyzed in both groups of patients.Data were analyzed with Chi-square test and logistic regression by using SAS software version 9.0.Results The incidence of UGB in DAT group was significantly higher compared with SAT group(7.1％ vs.3.4％,P ＜ 0.01).The factors,including DAT,age over 65 years,medication duration over 3 months and previous digestive diseases increased the risk of UGB in CHD patients(P ＜ 0.05),while the use of PPIs(proton-pump inhibitors)or gastric mucosal protectives lowered the risk of UGB(OR:0.415,95％ CI 0.226 ～ 0.762,P =0.0035).Conclusions DAT can significantly increase the risk of UGB.UGB can be increased in elderly CHD patients and those with long-term medication and previous digestive diseases.Utility of PPIs or gastric mucosal protectives can effectively lower the incidence of UGB.

[ ABSTRACT] AIM:To study the effects of anti-aging Klotho protein on neonatal rat myocardial cells with hypo-xia/reoxygenation ( H/R) injury.METHODS:The cardiomyocytes of neonatal SD rats were cultured to establish hypoxia/reoxygenation model.The myocardial cells were divided into normal control group, H/R model group, different concentra-tions of Klotho protein (0.1μmol/L, 1μmol/L and 10μmol/L) pretreatment groups.The myocardial cells pulse frequen-cy was observed before and after H/R.The cell viability was measured by MTT assay.The leakages of LDH, CK and AST, the content of MDA and the activity of SOD were detected.The apoptotic rate of the myocardial cells was analyzed by flow cytometry.The mRNA expression of endoplasmic reticulum stress markers and apoptosis-related molecules GRP78, CRT, CHOP and caspase-12 was measured by real-time PCR.The protein levels of CHOP, caspase-12 and phosphorylated Akt in the myocardial cells were determined by Western blot.RESULTS: Compared with normal control group, the pulse fre-quency, cell viability rate and SOD activity of myocardial cells were significantly decreased, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were increased in H/R model group.The mRNA expressions of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and caspase-12 were increased, whereas p-Akt level was decreased obviously.Compared with H/R model group, the pulse frequency, cell viability rate and SOD activity were in-creased significantly, the cell apoptotic rate as well as the contents of LDH, CK, AST and MDA were decreased in Klotho pretreated group.The mRNA expression of GRP78, CRT, CHOP and caspase-12 as well as the protein levels of CHOP and
caspase-12 were decreased, while p-Akt level increased significantly.CONCLUSION:Anti-aging Klotho protein improves the myocardial cell survival and inhibits the apoptosis by increasing the resistance of the cells to oxidative stress and exces-sive endoplasmic reticulum stress response, which is related with the activation of Akt phosphorylation in H/R-injured my-cardial cells.

Objective To investigate the efficacy of 50% acetic acid as a renal cyst sclerotherapy agent, and with further comparison to that of absolute alcohol. Methods Eighty five patients with renal cyst were undergone sclerotherapy through spiral CT guidance including 43 cases with absolute alcohol and the others with 50% acetic acid as sclerosing agents. All the cysts were aspirated under CT-guidance, beforehand. The sclerosising agents were withdrawn from the cysts after a definite period of retention. Results The disappearance rates of cyst cavity with absolute alcohol and acetic acid were 55.81% and 71.42%, respectively. Complication occurence rates with absolute alcohol and acetic acid were 16.28% and 4.76% , respectively. The average retention periods of absolute alcohol and acetic acid in cyst were (20?4)minutes, and (10?2)minutes, respectively. Statistical analysis demonstrated that all the data in two groups were significantly different. Conclusion Using 50% acetic acid as sclerosising agent in treating renal cyst possesses the better effect and less side effect, providing a tendency to replace the traditional therapy.

AIM:To observe the effects of neuroglobin ( NGB) overexpression on the apoptosis induced by Aβin the brains of double transgenic AD ( APPswe/PS1dE9) mice and to explore its potential mechanisms .METHODS:Twenty-four 13-month-old double transgenic AD mice were randomly divided into 3 groups:intracerebroventricular injection with normal saline (NS) group, intracerebroventricular injection with pcDNA3.1 and NS group, and intracerebroventricular injection with pcDNA3.1 and pNGB group.Immunohistochemistry was used to detect the expression of Aβ1-42 in the brains. TUNEL staining was used for analyzing the apoptosis , and the protein levels of cleaved caspase-3, caspase-9, PI3K, Akt and p-Akt were determined by Western blot .RESULTS: After intracerebroventricular injection with pNGB , the areas of Aβ1-42 in the hippocampus and cortex were decreased compared with NS group and pcDNA 3.1+NS group (P<0.01).The TUNEL-positive staining cells in the pNGB group were less than those in NS group and pcDNA 3.1 group ( P<0.01 ) . NGB overexpression attenuated the protein levels of cleaved caspase-3 and caspase-9 (P<0.01), but induced the produc-tion of PI3K and p-Akt (P<0.01).CONCLUSION:Overexpression of pNGB significantly inhibits the generation of Aβand attenuates the apoptosis induced by Aβ, indicating that NGB overexpression activates PI 3K/Akt pathway and inhibits the production of cleaved caspase-3 and caspase-9, which were tightly related with apoptosis .

Objective To evaluate the clinical value of Gli1 expression in breast cancer.Methods Immunohistochemistry was used to analyze the protein expression of Glil in breast cancer tissues,adjacent cancer tissues and metastatic lymph node.Results ①Breast cancer tissues expressed high levels of Gli1 compared to paracancerous tissues.Gli1 overexpression was found in 93.3％ of breast cancer tissues with recurrence and metastasis and 81.2％ in breast cancer tissues without metastasis.Gli1 expression could be detected either in interstitial tissues of cancer or in cancer tissues,or in both.The rate of Gli1 overexpression was 89.3％ in the inter stitial tissues of the primary cancer with recurrence and metastasis and 13.4％ in interstitial tissues of breast cancer tissues without metastasis.The difference had statistical significance(P ＜ 0.001).②High expression of Gli1 in breast cancer tissues was closely related to early recurrence and metastasis of breast cancer.Gli1 expression displayed a significant correlation with relapse-free survival (RFS) (P =0.046).The recurrence rate was 17.8％ in the high expression group in breast cancer tissue,whereas it was only 6.2％ in the low expression group(P =0.046),while the recurrence rate was 31.2％ in the high expression group in interstitial tissues,whereas it was only 11.1％ in the low expression group(P ＜ 0.001).Conclusion The high expression of Gli1 in breast primary cancer tissues,metastasis tissues and interstitial tissues is associated with recurrence and metastasis of breast cancer.

AIM:To explore the effects of transforming growth factor-α( TGF-α) in the monoclonal formation , proliferation, migration and adhesiveness of human endothelial progenitor cells ( EPCs).METHODS: The isolated and cultured EPCs were treated with various concentrations of TGF-α(final concentrations of 1, 5, 10μg/L, respectively).At the same time, the PBS control and epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) group (10μg/L TGF-αplus 1∶1 000 EGFR-TKI) were set.The effects of TGF-αon monoclonal formation , proliferation, migration and adhesiveness of EPCs were determined by clone formation experiment , thiazolyl blue tetrazolium bromide (MTT), EdU, Transwell and adhesion assays , respectively.The expression of epithelial growth receptor (EGFR) and vascular endothelial growth factor ( VEGF) were measured by Western blotting .RESULTS:Different concentrations of TGF-αall significantly induced the monoclonal formation , proliferation, migration and adhesiveness of EPCs (P<0.01), which were inhibited by EGFR-TKI.The results of Western blotting showed that TGF-αalso induced the expression of EGFR and VEGF with a cer-tain concentration effect ( P<0.01) .CONCLUSION:By combining with EGFR induced the expression of VEGF , TGF-αsignificantly promotes the related cell function of monoclonal formation , proliferation, migration, adhesiveness in EPCs.

AIM: To study the neuroprotective roles of neuroglobin (NGB) over-expression in the SH-SY5Y cells transfected with pAPPswe.METHODS:The plasmid pEGFP-NGB was successfully constructed and transfected into the SH-SY5Y cells, which were pretreated with pAPPswe.MTT assay was applied to detect the effect of NGB over-expres-sion on the cell survival rates.JC-1 staining was used to detect the level of mitochondrial transmembrane potential.The cell apoptosis was analyzed by flow cytometry.The effects of NGB over-expression on the protein level of p-Akt, Akt and caspase-3/9 were determined by Western blotting.The generation of Aβ42 in the cells was measured by ELISA.RE-SULTS:The cell survival rate was remarkably increased after transfection with NGB compared with control group and emp-ty plasmid group (P<0.05).The over-expression of NGB significantly inhibited the decrease in mitochondrial membrane potential induced by pAPPswe.The over-expression of NGB inhibited the apoptosis of the cells.Furthermore, over-expres-sion of NGB not only inhibited the expression of caspase-3 and caspase-9, but also induced the production of p-Akt, which was prevented by LY294002, an inhibitor of PI3K/Akt.The generation of Aβ42 was inhibited in the cells with the over-ex-pression of NGB.CONCLUSION: Over-expression of NGB significantly inhibits the SH-SY5Y cell injuries induced by pAPPswe and inhibits the expression of caspase-3/9, which is tightly related with cell apoptosis.Furthermore, the neuro-protective roles of NGB may be via activating PI3K/Akt signaling pathway.

Objective To study the appearance of MR diffusion weighted imaging in early stages of cartilage degeneration and to detect its values. Methods In 20 goat left knees, intra- articular injection of 5 units of papain was performed causing a loss of cartilage proteoglycan. Twenty right knees were used as control group. MR diffusion weighted imaging was performed at 24 hours after intra-articular injection of papain. ADC of each part of articular cartilage was measured and compared with each other. The proteoglycan content was measured biochemically and histochemicaUy. Routine MRI and DWI were performed in 100 patients with osteoarthritis and 20 healthy people. The ADC of each interested part of articular cartilage was measured and compared with each other. Results In experimental control group, the ADCav of articular cartilage was (14.2±2.3)×10-4 mm2/s. In early stages of cartilage degeneration group, the ADCav of articular cartilage was (17.5±4.2) × 10-4 mm2/s. The ADCav of the control group was lower than that of the early stages of cartilage degeneration group (t = 2.709 ; P = 0.016) . The proteloglyean content of articular cartilage was 4.22×10<'6> μg/kg in control group, and 0.82×10<'6>μg/kg in experimental group at 24 hours after injection of papain. The difference between control group and experimental group was significant (t = 2.705, P = 0.018). In healthy people, the ADCav of articular cartilage was (7.6±2. 2) × 10-4 mm2/s. In osteoarthritis group, the ADCav of articular cartilage was (10.3±4. 2) × 10-4 mm2/s. The ADCav in the healthy group was significantly lower than that in the osteoarthritis group (t = 2.609, P = 0.014). Conclusion DWI is an useful method in detecting early stages of cartilage degeneration which can not be showed on routine sequences.

Objective To evaluate the feasibility and effect of transthoracic echocardiography(TTE)on guiding the occlusion of the soft-rim atrial septal defect(ASD).Methods Sixty two patients with the soft-rim ASD were enrolled.The size of ASD was measured and rim of ASD was observed by TTE on various views by using color Doppler system with tissue harmonic function before occlusion,and filmy rim of ASD with flapping which could not sustain occluder was eliminated.The size of occluder was selected by integratively judging the size of ASD and"sustainable diameter of ASD"The waist size of occluder was measured after releasing occluder and compared with the longest diameter of ASD and"sustainable diameter of ASD"measured by TTE.Results The longest diameter of ASD measured by TTE before occlusion was 11-35 mm[average(21.6±5.2)mm],the "sustainable diameter of ASD"was 15-37 mm[average(25.6±5.(J)mm],the size of selected occluder was 18-44mm[average(30.7±5.5)mini and the waist size of released occluder was 13-35 mm[average(24.2±5.6)mm].Fine correlation was existed between the longest diameter of ASD measured by TTE and the waist size of released occluder(r=0.86,P＜0.000I).Morever,improved correlation was found between the"sustainable diameter of ASD"measured by TTE and the waist size of released occluder(r=0.89,P＜0.0001).Occluder was firmly fixed without falling in all patients.Conclusions TTE with tissue harmonic function can be used to measure the size of soft-rim ASD and the"sustainable diameter of ASD".It is a feasible,and effective method on guiding occlusion of soft-rim ASD.