[ 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.

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 study the quality of life(QOL) of patients with mild to moderate primary hypertension after treatment adjusted medication time by ambulatory blood pressure .Methods All 128 cases were divided into two groups randomly including chronos medication group and normal medication group .We tested ambulatory blood pressure ,QOL and influencing factors before medication and 6 months later .Results It′s observed the parameters of the ambulatory blood pressure improved significantly in all of the groups .There existed significant difference in nSBP ,nDBP ,nMAP ,SBP in dangerous night time ,and nocturnal blood pres‐sure decline rate between two groups (P<0 .05) .There existed very significant difference in DBP and MAP in dangerous night time (P<0 .01) .There were significant difference in the fields of MH (mental health) and RE (role limitation because of emotional health problem) (P<0 .05) ,and very significant difference in the fields of PF (physical functioning) ,RP (role limitation because of physical health) ,VT(vitality) (P<0 .01) .Multiple regression analysis indicate the influencing factors of the quality of life about patients with hypertension are age ,BMI index number ,medication time ,average family income ,medical insurance type ,culture de‐gree and job title .Age ,BMI index number and medication time depress QOL of patient with hypertension while average family in‐come ,better medical insurance type ,culture degree and job title improve it .Conclusion It′s possible to improved the parameters of the ambulatory blood pressure for patients with mild to moderate primary hypertension by chronos medication compared with nor‐mal medication more significantly .It′s useful to improve the QOL of patient with hypertension by improving average family income , medical insurance type and culture degree .

Objective To compare the quality of life before ischemia postconditioning and 6 months after ischemia postconditioning in patients with acute ST segment elevation myocardial infarction,and investigate the related risk factors.Methods One hundred and twenty-nine patients with acute ST segment elevation myocardial infarction were given the ischemie postconditioning.The quality of life was evaluated before ischemia postconditioning and 6 months after ischemia postconditioning by 36-item short-form health survey (SF-36) and seattle angina questionnaire (SAQ).Results In acute ST segment elevation myocardial infarction patients treated with ischemia postconditioning,the scores of SF-36 and SAQ were significantly lower at 6 months after ischemia postconditioning than those before ischemia postconditioning [SF-36:total score of body health (76.4 ± 17.3) scores vs.(56.3 ± 16.8) scores,physiologic function (75.3 ± 18.1) scores vs.(52.4 ± 19.2) scores,physiologic functional authority (75.6 ± 16.5) scores vs.(48.5 ± 20.3) scores,body pain (77.2 ± 15.4) scores vs.(58.7 ± 16.6) scores,total health state (73.6 ± 17.8) scores vs.(50.6 ± 14.7)scores,total score of mental status (77.5 ± 15.3) scores vs.(55.3 ± 17.3) scores,vitality (69.3 ± 18.1)scores vs.(43.2 ± 17.4) scores,society function (70.3 ± 17.5) scores vs.(41.3 ± 14.2)scores,affection functional authority (80.2 ± 17.4)scores vs.(63.5 ± 14.3)scores,mental health (77.6 ± 18.4) scores vs.(55.6 ± 20.1) scores,health change (76.3 ± 17.4) scores vs.(53.4 ± 16.6) scores;SAQ:body limitation of activity (78.46 ± 12.21) scores vs.(35.34 ± 15.33)scores,angina stable state (74.23 ± 8.53) scores vs.(30.12 ± 5.38) scores,angina episode state (72.34 ± 10.33) scores vs.(27.33 ± 9.12) scores,satisfaction degree of treatment (76.42 ± 12.13)scores vs.(30.56 ± 15.57) scores,knowledge of diseases (74.22 ± 9.35)scores vs.(37.25 ± 20.32) scores] (P ＜ 0.05).The results of multiple linear regression equation showed that the older,more severe vascular lesions,and lower total score of body health score before ischemia postconditioning indicated lower total score of body health,and worse quality of life after ischemia postconditioning.Conclusions The quality of life of patients with acute ST segment elevation myocardial infarction after ischemia postconditioning is significantly improved,compared with that before ischemia postconditioning.Advanced age,vascular disease and lower total scores of body health before ischemia postconditioning are independent risk factors of total scores of body health after ischemia postconditioning.

Object To develop an improved RP-HPLC method for determining ferulic acid in the human serum. Methods The determination was carried on RP-HPLC, using Kromasil-C 18column, methanol-water-acetic acid (36.4∶63∶0.6) as mobile phase with a flow rate of 1.0 mL/min and at the detection wavelength of 322 nm, and p-hydroxybenzaldehyde was used as internal standard. The serum sample was deproteinized with acetonitrile to extract ferulic acid. Results The calibration curve showed good linearity over the range of 9.94-159.04 ng/mL (r=0.992 5). RSD was less than 10% within day and day-to-day, the average recovery was 99.77% and the minimal concentration in serum was 5 ng/mL. Conclusion This method, which is simple, rapid, sensitive, reproducible and low toxic, is appropriate for the quantitative determination of ferulic acid in the human serum.

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.

Objective To investigate the therapeutic effect of quercetin on acute gouty arthritis and its influence in re-nal function in rats. Methods Seventy male Sprague-Dawley rats were randomly divided into normal control group, model group, colchicine group (0.5 mg/kg), allopurinol group (20 mg/kg), quercetin 100, 200 and 400 mg/kg groups (n=10 for each group). Rats were administered various drugs by oral gavage once a day for seven consecutive days throughout the experi-ment. On the fifth day, the animal model of acute arthritis was set up by giving monosodium urate crystal combined with hypo-xanthine. The inflammatory reaction was detected by measuring the circumference of right hind leg anklejoint with a tie line method at 0, 2, 6, 12, 24 and 48 h. The swelling ratio was calculated. The serum levels of uric acid (UA),β2-microglobulin (β2-MG), cystatin C (Cys-C), urea nitrogen (Urea) and creatinine (Cr) were detected by colorimetry and enzyme-linked im-munosorbent assay.Rats were sacrificed at the end of experiment, and the kidney was weighed and the renal index was calcu-lated. Results Treatment with quercetin, colchicine or allopurinol can significantly attenuate swelling rate in rats of acute gouty arthritis. The serum levels of UA were significantly higher in colchicine group and quercetin group than those of nor-mal control group and allopurinol group. The serum levels of UA was significantly lower in allopurinol group than that of nor-mal control group. After 48-h modeling, there was no significant difference in serum UA level between seven groups except allopurinol group. The levels ofβ2-MG and Cys-C were the lowest in normal control group than those of other groups. The se-rum levels of Urea and Cr and renal index were the highest in allopurinol group compared with those of other groups ( P<0.05). Conclusion Quercetin shows a significant effect of anti-inflammatory on acute gouty arthritis in rats. The model es-tablishment may lead to different degrees of renal damage. Quercetin has no protective effect against renal injury, and allopu-rinol aggravates kidney injury.

Objective To explore the effect of gestational hypertension on multiple organ system in neonates. Methods A total of 100 newborns whose mother had pregnancy complicating primary hypertension admitted to our hospital from December 2011 to December 2012 were selected and divided into preeclampsia group (n=53), gestational hypertension group (n=47) according to the blood pressure during pregnancy. Meanwhile, 100 newborns with healthy mother were selected as control group including 12 term infants and 88 premature infants. Data including birth weight, length and head circumference, Apgar score, the percentage of amniotic lfuid pollution, placental abruption and fetal distress, Neonatal Behavioral Neurological Assessment (NABA) score, serum level of creatine kinase, pulmonary arterial pressure, thyroid function, blood glucose, blood routine, cranial MRI parameters were collected and compared among three groups. Results In preeclampsia group, the gestational age, birth weight and head circumference, 1-min and 5-min Apgar scores were lowest while the ratio of low birth weight infants was highest among three groups, and the differences were signiifcant (P<0.05). In preeclampsia group, the rates of antenatal abnormalities (amniotic lfuid meconium III degree pollution, placental abruption and fetal distress) and complications (severe infection, myocardial damage, neonatal polycythemia, liver and kidney damage, hypoglycemia, hypothyroidism and respiratory failure) were highest among three groups, and the differences were signiifcant (P<0.01). In preeclampsia group, the red blood cell count, the levels of hemoglobin, hematocrit and thyroid stimulating hormone were signiifcantly higher than those in the other two groups ( P<0.05 ), and the white blood cell and platelet count was signiifcantly lower than that in the control group (P<0.05). The passive muscle tension scores in preeclampsia group were signiifcantly lower than those in the other two groups (P<0.05).The abnormity rate of cranial MRI in preeclampsia group was highest among three groups, and the difference was signiifcant (P<0.01). Conclusions Gestational hypertension may cause multisystem disorders in newborns, such as fetal intrauterine growth restriction, endocrine system disorders, heart dysfunction, increased blood viscosity, delayed neurodevelopment. The severity of gestational hypertension is associated with the adverse impact on the multiple systems in neonates.

Objective To explore the level and the significance of amino-terminal propeptide of procollagen type Ⅲ(P Ⅲ NP)in the serum of hypertrophic scar patients at excessive stages.Methods The serum samples of 74 inpatients admmitted in Affiliated Fosham Hospital of Sun Yat-sen University from August 2007 to August 2009 suffering from long-persisting post-burn hypertrophic scar at various stages were collected.Serum samples of 12 were normal persons were also collected and used as control group.Hypertrophic scar group were divided into 4 subgroups according to the phase of scar and the serum concentrations of P Ⅲ NP were detected using the sensitive RIA method.Results The level of PⅢ NP increases gradually during the process of immature hypertrophic scar to mature scar before it peaking in 4 to 6 months scar subgroup.The concentration of PⅢ NP degreased gradually with the maturation of hypertrophic scar.Conclusions The high concentration of PⅢNP can reflect the high level of the synthesis of type Ⅲ collagen in the tissue of post-burn hypertrophic scar.The level of P Ⅲ NP is synonymous with the ongoing process of hypertrophic scar.PⅢ NP may be a satisfactory marker in discerning the growth and development of post-burn hypertrophic scar.