Telmisartan reduces retina vessel endothelial cell apoptosis via upregulating retinal ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of angiotensin II (Ang II) antagonist telmisartan on retina vessel endothelial cell apoptosis and its impact on the ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats (SHR).</p><p><b>METHODS</b>Thirty-six SHR 16 week-old were randomly divided into 3 groups (n = 12 each): SHR, SHRT (telmisartan 10 mg · kg-1 · d-1 by gastric gavage) and SHRTA group (telmisartan 10 mg · kg-1 · d-1 by gastric gavage plus intravenous injection of A-779 0.5 mg · kg-1 · d-1), twelve WKY rats served as normotensive control group. Systolic blood pressure was measured at pre-treatment and 8 weeks later. After 8 weeks, rats were sacrificed, the expression of ACE2 and Mas in retina were analyzed by qRT-PCR, Western blot and Immunohistochemistry, the Ang-(1-7) concentration in serum was measured by ELISA. Specimens were obtained and stained by hematoxylin and eosin, and the morphology of retina vessel was observed. Apoptosis of vessel endothelial cells were determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method.</p><p><b>RESULTS</b>The systolic blood pressure of SHR, SHRT and SHRTA groups at baseline were significantly higher than age-matched WKY group (all P < 0.01). Eight weeks later, the systolic blood pressure group was significantly lower in SHRT group than in the SHR group (P < 0.01), this effect was partly reversed in SHRTA group. The retinal ACE2 mRNA and protein expression was significantly lower in SHR group than in WKY and SHRT groups (P < 0.01), which was similar between SHRT group and SHRTA group (P > 0.05). The retinal Mas mRNA and protein expression were significantly lower in SHR group compared to WKY and SHRT groups (all P < 0.01), which was significantly lower in SHRTA group than in the SHRT group (P < 0.05). ELISA results showed that serum Ang-(1-7) protein level was significantly lower in SHR group than in WKY group and SHRT group (both P < 0.05), which was lower in SHRTA group compared to SHRT group. Retinal vessel endothelial cell apoptosis was higher in SHR group than in WKY group, which could be reduced by cotreatment with telmisartan and this beneficial effect could be reversed by A-779.</p><p><b>CONCLUSION</b>Telmisartan can reduce retinal vessel endothelial cell apoptosis via upregulating the ACE2-Ang-(1-7)-Mas axis.</p>

Analysis of related factors for pacemaker pocket infection in elderly patients after implantation of permanent pacemakers / 中华老年医学杂志

Objective To analyze related factors for pacemaker pocket infection in elderly patients after implantation of permanent pacemakers and to provide a theoretical basis for preventing pacemaker pocket infection.Methods Pacemaker pocket infection and related factors were analyzed for 412 patients who received implantation of permanent pacemakers from Apr.2010 to Jun.2013 in the Department of Cardiology.Results With 5 cases of pacemaker pocket infection,the rate of infection was 1.2％.The infected patients were older than the uninfected patients [(74.5±4.2) years vs.(60.3±6.6) years,t=4.781,P＜0.01].The rate of infection was higher in patients who had undergone operations twice or more than in patients who had undergone one operation [10.0％ (3/300) vs.0.5％ (2/382),x2=10.583,P＜0.01].The rate of infection was higher in patients with the operation lasting 2 hours or longer than in patients with the operation time shorter than 2 hours [(3.8％ (4/106) vs.0.3％ (1/306),x2=7.802,P＜0.01].The rate of infection was higher in patients with pocket hematoma than in patients without pocket hematoma [16.7％ (3/18) vs.0.5％(2/394),x2=37.492,P＜0.01].Independent risk factors for pacemaker pocket infection included pocket hematoma (OR=6.193),number of operations≥2 (OR=2.594),operating time≥2 hours (OR=2.265) and age of 75 years or older (OR =2.193).Conclusions Pocket infection after implantation of permanent pacemakers is related to pocket hematoma,number of operations,operating time and age.

Effect of siRNA interfering SLC7a8 on uptaking L-dopa in NRK-52E cells / 中国病理生理杂志

AIM: To investigate the effect of selective silencing of SLC7a8 on uptaking L-dopa in renal tubular epithelial cells of rat (NRK-52E). METHODS: The three siRNAs targeting SLC7a8 (siRNA-1, siRNA-2, siRNA-3) were designed and synthesized. A siRNA with nonspecific coding sequence (siRNA-con) was used for control. All siRNAs were transfected into NRK-52E cells. The siRNA-con transfected group, blank control group and gene-specific silencing SLC7a8 group were set up. The efficiency of transfection was estimated by flow cytometry. The efficiency of RNA interference was detected and screened by RT-PCR preliminarily, and was followed by Western blotting at protein level. The concentrations of L-dopa uptake into the NRK-52E cells were detected by ultraviolet spectrophotometry at different time points (6, 12, 24, 36, 48, 60, 72 and 120 min). RESULTS: The transfection efficiency was 94% detected by flow cytometry. The initial screening of RT-PCR showed that the efficiencies of RNA interference of siRNA-1 and siRNA-3 were higher, and siRNA-3 was the highest at protein level determined by Western blotting. No distinctive change was found between siRNA-con treated NRK-52E and blank control cells. The L-dopa uptake at different time points (6, 12, 24, 36, 48, 60, 72 and 120min) in siRNA-interference group was lower than that in siRNA-con transfected group and blank control group. No significant difference of L-dopa uptake between siRNA-con group and blank control group was observed. CONCLUSION: RNA interference technology selectively down-regulates SLC7a8 expression in rat renal tubular epithelial cells. The L-dopa uptake is also decreased after specifically silencing the slc7a8 expression.

Up-regulation of Bcl2l14 gene in myocardium of Pax-8 gene knockout mouse / 中国病理生理杂志

AIM: To investigate the downstream genes of the transcriptional factor Pax-8 related to cardiopathy. METHODS: The total RNA derived from the heart of Pax-8 KO~(-/-) and Pax-8 KO~(+/-) mice was extracted. Mouse genome DNA microarray containing 31 802 mouse oligonucleotides probes was used to investigate the differential expression between the Pax-8 KO~(-/-) and the Pax-8 KO~(+/-) mice hearts. The candidate genes were confirmed by RT-PCR and real time RT-PCR assay. RESULTS: Microarray results showed that, compared to the Pax-8 KO~(+/-) mice, 25 genes were down-regulated and 17 were up-regulated in the Pax-8 KO~(-/-) mice, concerning metabolize enzymes, cell signal conducting and nuclear transcript factors and so on. Bcl2-like 14 (Bcl2l14) was proved to be up-regulated by RT-PCR. Real time RT-PCR results revealed that Bcl2l14 in the Pax-8 KO~(-/-) mice was 2.07 and 2.23 fold as much as that in the Pax-8 KO~(+/-) and the Pax-8KO~(+/+) mice (P<0.01). CONCLUSION: The Bcl2l14 gene is one of the downstream genes of Pax-8 and probably plays an important role in the mechanism of ventricular septum defect.

Apoptosis of retinal capillary cells and expression of p53 in rats with spontaneous hypertension after ischemic reperfusion injury / 中华眼底病杂志

Objective To investigate the effect of apoptosis-related gene p53 on the apoptosis of retinal capillary cells in rats with spontaneously hypertensive (SHR) after ischemic reperfusion injury. Methods A total of 60 SHR rats were randomly divided into sham group (SHR-SH) and retinal ischemie reperfusion group (SHR-RIR), which were subdivided into 5 subgroups according to the time after RIR: 2, 6, 24, and 72 hours and 7 days, with 6 rats in each subgroup. Another 60 Wistar-Kyoto (WKY) rats were divided into the same groups as the SHR rats as the control. The RIR model was set up. The apoptosis of retinal capillary cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods and the expression of p53 was determined by streptavidin-perosidase (SP) immunohistochemistry. Results The apoptosis rate of retinal capillary cells in the 5 SHR-RIR groups was (8.64±0.56)%, (14.92±0.99)%, (24.72±2.98)%, (16.53±1.80)%, and (7.12±1.10)%,respectively. The expression of p53 in SHR-RIR groups increased at the 2nd hour after RIR, reached the peak at the 24th hour, kept the high level at the 72nd hour, and remained a little at the 7th day, which was significantly different from which in the SHR-SH groups (P<0. 01). The expression of p53 were higher in SHR-IR groups than that in the WKY-RIR groups (P<0. 01). Conclusions p53 may play a part in RIR injury by inducing or promoting apoptosis. The apoptosis of retinal capillary cells after RIR is more severe under the hypertension, and reaches the peak at the 24 hour after RIR.

Prognostic value of cardiac troponin I and CRP in patients undergoing percutaneous coronary intervention / 中华检验医学杂志

Objective,This study was designed to evaluate the predictive value of pre-procedure Cardiac troponin I(cTnI)and CRP levels in patients with acute coronary syndrome(ACS)undergoing percutaneous coronary intervention(PCI).Methotis cTnI and CRP were determined on admission in 335 consecutive patients with ACS who underwent primary PCI.Blood samples were obtained within 6-10 h after onset of symptom.The concentration of cTnI was determined by an automated chemiluminescence immunoassay.CRP was measured by immunoassay assay.According to the admission cTnI(＜0.1,0.1-0.5,＞0.5μg/L)and CRP(≤3,＞3 mg/L)divided into different groups.The pre-procedure cTnI and CRP status associated with 30 days cardiac mortality and major adverse cardiac events(MACE.including cardiac death.non-fatal recurrent MI.heart failure.readmission for any reason)were analyzed.The cardiac mortality at follow.uD period of 2 years were analyzed.Results Muhivariate logistic regression analyses revealed preoperative cTnI predicted 30 days cardiac mortality(OR=3.5,95%CI 2.2-5.3,P＜0.01),and recurrent MI rate(OR=1.5,95%CI 1.1-2.6,P＜0.05),independent of other known prognostic factors such as age,gender,hypertension,Hypercholestemlemia,diabetes and smoking.The pre-procedure CRP was independently related to 30 days cardiac mortality(OR=1.6,95%CI 1.1-2.3.P＜0.05),whereas there was no relationship to the MI rate.In ACS,levels of CBP≤3 mg/L,the three different risk groups (cTnI＜0.1,0.1-0.5,＞0.5μL)with corresponding 30 days MACE rates of 4.3%,11.7%,18.8%(X~2=4.829,P=0.028),CRP＞3 mg/L,the three groups mth corresponding 30 days MACE mtes of 5.5%,13.2%,21.1%(X~2=5.862,P=0.015),respectively.Patients were followed up for 2 years,Kaplan-Meier survival analysis demonstrated a significantly reduced survival at 2 years in patients witll a cTnI ＞0.5μg/L(80.0%versus 89.1%for a cTnI of 0.1-0.5μg/L and versus 92.2%flor cTnI＜0.1μg/L;X~2=7.571,P＜0.05 by log-rank).Conclusions The levels of CRP and cTnI in Acs of onset in 6-10 h provide an even better risk stratification after the PCI.and closely correlate with 30 days MACE.Elevated cTnI provides long-term prognostic information regarding cardiac mortality.Therefore.The combination of CRP and cTnI measurement should be taken into consideration for risk stratification to decide about the management strategies in ACS patients.

Effect of Tetramethylpyrazine on the Function of Endothelial Progenitor Cells in vitro / 浙江中医药大学学报

[Objective] This study was to investigate the effect of TMP on function of EPCs in normal or hydrogen peroxide(H2O2)inducing injury model cell culture,and to approach a therapy to coronary heart disease(CHD).[Method] Total mononuclear cells were isolated from the cord blood by ficoll density gradient centrifugation,cultured in vitro.After the cells cultured for 7 days,identified them by cellmarkers and the ability of intaking the ac-LDL,and made sure they were EPCs.To divide the cells into three groups,TMP(5,25,100,200mg),H2O2(500??)model and TMP together with H2O2,cultured for 24h and then detected the function and apoptosis of EPCs.[Result] Versus to control group,the TMP(5,25,100mg)groups had no important effect on the proliferation and adhesion of EPCs,but TMP(200mg)group had negative effect on EPCs.Versus to H2O2 model group,TMP(5,25,100mg)preincubated groups had important protective effect on EPCs in proliferation,adhesion and apoptosis.[Conclusion] TMP could protect EPCs from oxidative stress injury,but had little benefit to EPCs in normal incubation.

Preliminary study of endothelial progenitor cells formed vessel-like structure in vitro / 中国病理生理杂志

AIM: Endothelial progenitor cells(EPCs) are a group of stem cells/progenitor cells,which exist in postnatal body and can be of specially homing to the foci of angiogenesis and then differentiate into endothelial cells.This investigation was to study the method for culturing endothelia progenitor cells(EPCs) in vitro,and to observe its feasibility and condition formed vessel-like structure.METHODS: The cells were isolated from born marrow,peripheral blood,umbilical cord blood or spleen in different laboratories.The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro through adhesion selection and were differentiated into endothelial cells under the induction of special cytokines.The expression of CD34,VEGFR-2,AC133 and VE-cadherin were detected by fluorescence-activated cell sorting.The endothelial cell lineage was confirmed by DiI-ac-LDL up-taking and Ⅷ factor immunocytochemistry.RESULTS: The EPCs derived from human umbilical cord blood and rabbit peripheral blood were cultured in vitro successfully,forming cord-like and tube-like structure.The EPCs derived from rabbit peripheral blood differentiated more mature and formed vessel-like structure.CONCLUSION: The EPCs derived from human umbilical cord blood and rabbit peripheral blood formed vessel-like structure in vitro.EPCs may be a potential resource of vessel tissue engineering.

Protein tyrosine kinase of focal adhesion kinase subfamily-a new related gene of ventricular septal defect / 中国病理生理杂志

AIM: To investigate the function of the prot ein tyrosine kinase of focal adhesion kinase subfamily gene, the gene regulation and the signaling pathway related to ventricular septal defect during heart dev elopment. METHODS: The ALK3 downstream genes were screened usin g microarray. The ALK3 downstream genes was identified using RT-PCR and real tim e quantitative RT-PCR.RESULTS: It was found that the protein ty rosine kinase of focal adhesion kinase subfamily gene was up-regulated 3.7 fold in the 11.5 days embryonic heart of cardiac- specific deletion of ALK3 mice. CONCLUSION: The protein tyrosine kinase of focal adhesion kinase subfamily possibly is a regulatory factor in bone morphogenetic protein signalin g pathway and may be related with the development of ventricular septal defect.