Objective To study the effect of overexpression of tumor necrosis factor-α (TNFα) in failing heart on contractile function of mouse diaphragm. Methods Diaphragms excised from transgenic mice with cardiac specific overexpression of TNF-α and from wild-type littermate controls were studied in vitro with direct electrical stimulation. Cytosolic oxidant levels were measured with 2',7'-dichlorofluorescin diacetate (DCFH-DA). Emissions of the oxidized product were detected by fluorescence microscopy. Results The diaphragm contractile force in transgenic animals with heart failure was 47% less than in controls [(13. 2±0. 8) N/cm2 vs. (25.1±0. 6) N/cm2 , P＜O. 01]. The weakness was associated with greater intracellular oxidant levels (P＜0. 05) and was partially reversed by 30-minute incubation with the antioxidant N-acetylcysteine (NAC) 10 mmol/L ( P ＜ 0. 01 ).Exogenous TNF-a 500 mmol/L increased oxidant production in diaphragm of wild-type mice and caused weakness that was inhibited by NAC. Conclusions Overexpression of TNF in failing heart causes oxidative stress which leads contractile dysfunction in mouse diaphragm.

【Objective】 To study the removal effect of fibronectin(Fn) from von willebrand factor(vWF) by ion-exchange chromatography through processing human coagulation factor Ⅷ chromatographic washing products, in order to select a method that can effectively reduce Fn without compromising the activity yield. 【Methods】 In a multi-batch process development experiment, Fractogel^® EMD TMAE(M) strong anion filler produced by Merck(Germany) was used to conduct chromatography to investigate vWF ristomycins titer (vWF: RCof), vWF recovery, protein content and Fn content. 【Results】 During the development of vWF pilot purification process, the content of Fn in the samples can be effectively reduced by ion-exchange chromatography, with removal rate more than 87%, titer recovery of vWF more than 80%, and no significant change in other quality indexes. 【Conclusion】 The use of ion-exchange chromatography to purify vWF can effectively reduce the content of Fn, which has positive significance for developing new product process and improving the product quality of blood products manufacturers.

Objectives To study the predictive value of Toll-like receptors 3,4(TLR3,TLR4),fructosamine(FMN)and glycated hemoglobin A1c(HbA1c)in the in-stent restenosis and re-occlusion after primary percutaneous coronary intervention(PCI)in patients aged 70-85 years with old myocardial infarction.Methods 51 patients aged 70-85 years with in-stent restenosis after primary PCI from Jan 2007 to Sep 2016 were selected.Serum level changes in TLR3,TLR4 were detected by flow cytometry.The levels of FMN and HbA1c were tested by colorimetric endpoint reaction and high-pressure liquid chromatography respectively.Results The levels of TLR3,TLR4,FMN and HbA1c were gradually elevated along with the increases of artery numbers(0,1,2,>2)and percentage(0%,70-89%,90-99%,100%)of in-stent restenosis,LVEF(%)decrease and NYHA(Ⅰ,Ⅱ,Ⅲ,Ⅳ)increase(all P<0.01).The examples of data were selected in the following groups of artery numbers(0,1,2,>2)of in-stent restenosis in TLR3,and group of percentage(0%,70-89%,90-99%,100%)in the in-stent restenosis in TLR4,group of LVEF(%)in FMN,and group of NYHAⅠ,Ⅱ,Ⅲ,Ⅳ in HbA1c(%)(all P<0.01).The levels of TLR3(%)in artery numbers of restenosis(0,1,2,>2)groups were(7.6±0.5),(18.9±0.6),(32.0±0.9),(51.3±0.8),respectively(all P<0.01).The levels of TLR4(%)in the in-stent restenosis percentage(0%,70-89%,90-99%,100%)groups were(10.5±7.0),(20.1±7.2),(33.3±9.7),(69.0±11.3%)respectively(all P<0.01).The levels of FMN(mmol/L)in LVEF[(49~59%),(37~48%),(25~36%)]groups were(0.6±0.4),(9.4±0.6),(18.1±0.8),respectively(all P<0.01).And the level of HbA1c(%)in groups of NYHA Ⅰ,NYHA Ⅱ,NYHA Ⅲ,NYHA Ⅳ were(6.1±0.4),(5.9±0.6),(8.9±0.9),(12.0±0.8),respectively(all P<0.01).Conclusions Serum level changes in TLR3,TLR4,FMN and HbA1c may become the new indicators to forecast the degree of in-stent restenosis in very old patients with old myocardial infarction after primary coronary intervention.

Objective To study the relationship of levels of circulating endothelial microparticles (EMP62E, EMP31) and high-sensitivity C-reactive protein (hs-CRP) with severity of chronic left heart failure in elderly patients. Methods According to New York Heart Association (NYHA) class and left ventricular ejection fraction (LVEF), the healthy subjects and the patients were divided into five groups: control group [LVEF: (63.97±4.65)%], classⅠ group [LVEF: (42.67±2.06)%], classⅡ group [LVEF: (34.26±3.17)%], class Ⅲ group [LVEF: (29.05±1.07)%] and class Ⅳ group[ LVEF:(25.17±1.42)%] . The levels of circulating EMP62E, EMP31 and hs-CRP of the patients and healthy subjects were measured by flow cytometry and nephelometry immunoassay, respectively. Results There were significantly differences in EMP62E, EMP31 and hs-CRP between class Ⅳ group and classⅠ group P＜0.01) EMP62E [(1092.7 ± 102.8) counts/μl vs. (291.0±21.9) counts/μl], EMP31 [(1596.1±46.3) counts/μl vs. (477.8±40.3) counts/μl] and hs-CRP [(14.74±0.07) mg/L vs. (4.86 ± 0.09) mg/L]. The levels of circulating EMP62E, EMP31 and hs-CRP were gradually elevated significantly along with the increased severity of chronic left heart failure in elderly. Conclusions The upregulation of circulating EMP31, EMP62E and hs-CRP may contribute to the development of chronic heart failure in elderly.

Objective To observe the effect of changes in CD34+ cell level on various degrees of chronic heart failure (CHF) in the elderly. Methods The enrolled patients were divided into four CHF groups according to the New York Heare Association(NYHA) functional class: NYHA class I (n=23), Ⅱ (n=27), Ⅲ (n = 20) and IV group (n= 16) , and there were 41 healthy controls over the same period. The levels of peripheral blood CD34+ cells were measured, and the tumor necrosis factor-α (TNF-α). its soluble receptors (sTNFR-1 and sTNFR-2) and vascular endothelial growth factor (VEGF) were also measured. Results The levels of CD34+ cells were elevated in the early CHF and depressed in the advanced CHF in elderly patients. The levels of CD34+ cells were (0. 6± 0.2) 109/L in control group, (2.4±0. 4) 109/L in NYHA class I group, (1.9±0.2)×109/L in NYHA class Ⅱ group, (1.3±0.1)×109/L in NYHA class Ⅲ group and (0.5±0.2)×109/L in NYHA class Ⅳ group, respectively (all P<0.01).And TNF-α, sTNFR-1, sTNFR-2 and VEGF were increased in severe chronic left ventricular failure CNYHA class IV vs. I group:TNF-α: (61.4± 15.7) ng/L vs. (28.4±10.8)ng/L; sTNFR-1: (2820.9±1282.8)ng/L vs. (690.8±62.7) ng/L; sTNFR-2: (4113.1±1102.2) ng/L vs. (740.8± 112.3)ng/L; VEGF: (996.3±487.1)ng/L vs. (423. 3±147. 9)ng/L, all P<0. 013. Conclusions The changes of CD34+ cell level may predict various degrees of chronic heart failure in elderly patients.

Objective To study the changes in expression of high-sensitivity C-reactive protein (hsCRP),glutathione S-transferase Pi(GSTPi)and annexin A5 (AnxA5)in elderly patients with old myocardial infarction and the clinical significance.Methods Serum levels of GSTPi and AnxA5 were measured by ELISA and the level of hs-CRP was measured by immunoturbidimetry in elderly patients with old myocardial infarction (n =185)from December 2012 to November 2015.Results Along with the increasing coronary artery stenosis,GSTPi level was decreased and AnxA5/hs-CRP levels were increased in elderly patients with old myocardial infarction.In comparison between coronary artery stenosis ＞ 95％ group versus stenosis of 55％-65％ group,GSTPi was(190.0±37.0)μg/L vs.(289.0 ±86.0)μg/L,AnxA5 was(33.9±4.0)μg/L vs.(8.1 ± 2.9) μg/L,and hs-CRP was (15.3 ± 1.3) mg/L vs.(5.9 ± 0.8) mg/L with statistically significant differences(all P＜0.01).There were significant differences between LVEF 30％ group[GSTPi(198.0±39.0) μg/L,AnxA5(38.9±5.1)μg/L and hs-CRP(17.9± 1.9)mg/L]and LVEF 40％-54％ group[GSTPi(219.0± 61.0)μg/L,AnxA5 (12.9±3.9)μg/L and hs-CRP(10.1 ± 1.0) mg/L] (all P＜0.01).There were significant differences between NYHA Ⅳ group [GSTPi (171.0 ± 43.0) μg/L,AnxA5 (18.1 ± 5.0) μg/L and hs-CRP (16.9±2.1)mg/L]and NYHAⅠgroup[GSTPi(295.0±91.0)μg/L,AnxA5(7.3±3.1)μg/L and hs-CRP (7.8± 1.3)mg/L](all P＜0.01).Conclusions The expression of GSTPi,AnxA5 and hs-CRP in elderly patients with old myocardial infarction may become the new indicators to forecast the degrees of coronary artery stenosis and heart failure.

Objective:To establish a method for the determination of asperosaponinⅥin Dieda Cuyu tablets by HPLC. Methods:A Hypersil C18(250 mm ×4.6 mm,5 μm)column was used. The mobile phase was acetonitrile-water(30∶70) with a flow rate of 1.0 ml·min-1 . The detection wavelength was 212 nm, the column temperature was room temperature,and the injection volume was 10μl. Results:AsperosaponinⅥ showed a good linear relationship within the range of 0. 04-0. 32 μg(r=0. 999 6). The average recovery was 97. 84%(RSD=1. 70%, n=6). Conclusion:The method is simple,accurate and reproducible, which can be used in the deter-mination of asperosaponinⅥ in Dieda Cuyu tablets.

Objective:To investigate clinical characteristics for in-stent reocclusion lesions after coronary stent implantations in aged patients.Methods:229 patients diagnosed with chronic total reocclusions were recruited from Jan 2005 to Dec 2019 in this retrospective study.According to age, patients were divided into a 40-49 year-old group(n=60), a 50-59 year-old group(n=58), a 60-69 year-old group(n=55), and a 70-80 year-old group(n=56)to examine different lesion characteristics after coronary stent implantations.Results:In the 40-49 year-old group, the 50-59 year-old group, the 60-69 year-old group and the 70-80 year-old group, the rates of multi-vessel reocclusions were 11.6%, 15.5%, 21.8% and 25.0%, respectively( χ2=10.03, P=0.01). For each group, lesions with concurrent proximal and middle coronary reocclusions accounted for 8.3%, 12.0%, 30.9% and 35.7%, respectively( χ2=11.83, P=0.005); Reocclusions with severe coronary calcification accounted for 6.6%, 15.5%, 36.3% and 37.5%, respectively( χ2=11.56, P=0.006); Long coronary reocclusion lesions(36-47 mm)accounted for 15.0%, 17.2%, 21.8% and 25.0%, respectively( χ2=11.56, P=0.007); Coronary reocclusions with diffuse long calcified lesions accounted for 8.3%, 13.7%, 32.7% and 35.7%, respectively( χ2=10.80, P=0.01). Conclusions:The clinical characteristics of in-stent reocclusion lesions after coronary stent implantations include multiple chronic total coronary reocclusions, concurrent proximal and middle coronary reocclusions, heavily calcified coronary reocclusions, long coronary reocclusions and diffuse long calcified coronary reocclusions in aged patients.

Objective To study the clinical value of anti-oxidative stress biomarkers for diagnosing in-stent restenosis and in-reocclusion after coronary stent implantation in aged patients.Methods A total of 72 advanced-aged patients with in-stent restenosis and in-stent reocclusion after coronary stent implantation were successively recruited in this retrospective study from February 2010 to November 2017.Changes in serum superoxide dismutase 3(SOD3),nitric oxide(NO),endothelial cell nitric oxide synthase(eNOS)and malondialdehyde(MDA)levels were measured.Results Serum 1evels of SOD3,NO and eNOS decreased and serum MDA levels were elevated in advanced-aged patients with in-stent restenosis.There were significant differences in serum levels of SOD3,NO,eNOS and MDA between the advanced-aged patients without in-stent restenosis and the advanced-aged patients with multivessel in-stent restenosis or reocclusion[(20.0±3.2) × 103U/L vs.(10.9±3.9) ×103U/L,(61.2±14.2)μmol/L vs.(28.3±17.2)μmol/L,(75.9±24.7)ng/L vs.(33.0±119.6)ng/L,(2.2±1.4)nmol/L vs.(11.7±3.1)nmol/L,respectively,P＜0.01].Patients with 50-69％ restenosis had higher serum levels of SOD3,NO and eNOS and lower levels of MDA than patients with 100％ restenosis[(21.3 ± 2.9) × 103 U/L vs.(10.3 ± 4.0) × 103 U/L,(59.7 ± 16.7) μmol/L vs.(38.3 ±16.3)μmol/L,(74.5±21.1)ng/L vs.(41.9±26.8)ng/L,(2.6±3.9 nmol/L)vs.(10.1±3.1)nmol/L,respectively,P ＜ 0.01].Patients with left ventricular ejection fraction (LVEF) ≥ 55 ％ had higher serum levels of SOD3,NO and eNOS and lower levels of MDA than patients with LVEF＜30％ [(21.0±4.1) × 103 U/L vs.(5.3±1.9) × 103 U/L,(60.1 ± 14.2)μmol/L vs.(29.0± 13.2)μmol/L,(74.7±25.1)ng/L vs.(39.3 ± 20.3) ng/L,(2.3 ± 1.5) nmol/L vs.(10.0 ± 3.9) nmol/L,respectively,P ＜0.01].Serum levels of SOD3,NO and eNOS were higher and MDA levels were lower in patients with New York Heart Association(NYHA)Class Ⅰ than in patients with NYHA Class Ⅳ[(22.1±3.5)×103U/L vs.(9.7±2.9) × 103 U/L,(62.9± 13.9)μmol/L vs.(24.9± 13.3)μmol/L,(76.7±26.7) ng/L vs.(41.9±21.5)ng/L,(2.7± 1.9)nmol/L vs.(8.7±3.8)nmol/L,respectively,P＜0.01].Conclusions Serum level changes of anti-oxidative stress biomarkers such as SOD3,NO and eNOS may have clinical value in diagnosing in-stent restenosis and in-reocclusion after coronary stent implantation in aged patients.

Objective To investigate risk factors for in-stent restenosis and reocclusion after coronary stent implantation in aged patients.Methods 131 patients diagnosed with chronic total occlusion and old myocardial infarction due to coronary stenosis were recruited in this retrospective study from Mar 2004 to May 2015.Patients were divided into 50 to 59 years old group (n=51),60 to 69 years old group (n=43),and 70 to 80 years old group (n=37) to study coronary lesion characteristics.In-stent restenosis and reocclusion were detected at 6,12,18,and 24 months after coronary stent implantation.Results Before coronary stent implantation,the incidence rate of type 2 diabetes was significantly increased with three increasing age groups:9.8％ at ages 50-59 group (n=5),18.6％ at ages 60-69 group (n=8),and 27.0％ at ages 70-80 group (n=10) (all-P＜0.01).The incidence rates of multiple coronary artery disease,long coronary lesions (＞20 mm),eccentric coronary lesions,serious angle of coronary lesions,irregular coronary lesions,proximal coronary curvature,moderate to severe calcified coronary lesions,coronary restenosis (90％-99％ or 100％),and complex bifurcation lesions were significantly elevated with three increasing age groups (P ＜0.01 or P ＜0.05).The ratios of patients with in-stent restenosis at 24 months after coronary stent implantation were significantly elevated with three increasing age groups:at 9.8％ (n=5),18.6％ (n=8),and 27.0％(n=10) for 90％ 99％ restenosis sub-group,and at 5.9％ (n=3),14.0％ (n=6) and 24.3％ (n=9) for 100％ restenosis sub-group,respectively (all P＜0.05 or P＜0.01)Conclusions Type 2 diabetes is an independent risk factor for complex coronary lesions in aged patients Complex coronary lesions,three or more stents,and long coronary stents may lead to ir-stent restenosis and reocclusion after coronary stent implantation in aged patients.