Application of transcatheter aortic valve replacement in elderly patients with native pure aortic regurgitation / 中国医师杂志

Objective:This study sought to investigate the feasibility, anatomical indications and operating points of transcatheter aortic valve replacement (TAVR) procedure in the treatment of pure aortic regurgitation (AR).Methods:The medical records of 4 elderly patients with pure AR who were treated in the cardiology department, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from March 2020 to March 2021 were retrospectively analyzed. All patients were implanted with self-expandable valve stents via peripheral artery approach for TAVR treatment. The feasibility, anatomical indications and key points of TAVR were analyzed.Results:The 4 patients with pure AR who were carefully screened had an average age of 66 years, and all achieved TAVR treatment success without serious perioperative complications and death. Postoperative examination and follow-up data showed that cardiac functions and cardiac remodeling indexes were significantly improved.Conclusions:This exploratory study shows that TAVR is technically feasible and effective treatment option for selected elderly patients with native pure AR, which is worthy of further study.

Renal artery stenosis in children after pediatric donor kidney transplantation: a report of five cases / 中华器官移植杂志

Objective:To explore the clinical features of transplanted renal artery stenosis after pediatric donor kidneys in children.Methods:We retrospectively summarized the clinical data in five cases of transplanted renal artery stenosis undergoing deceased pediatric donor kidney transplantation from May 1, 2014 to June 30, 2021.Donor/receptor characteristics, diagnosis, treatment and prognosis were recorded.The median follow-up period was 29 months.The median age of five donors and recipients was 9 and 132 months respectively.En bloc renal allograft( n=2)and single kidney transplantation( n=3)were performed.End-to-side anastomosis was performed for renal arteries.The median diagnostic time of renal artery stenosis was 10(3-60)months post-transplantation.Except for one 3-year-old recipient with an earlier onset of stenosis, four stenotic cases during a rapid growth period had a maximal height increase of 30 cm post-transplantation.Three of them had a history of surgery at graft site, including previous kidney transplantation( n=1)and transplant urinary tract reconstruction( n=2). All five cases had hypertension and two showed an elevated serum level of creatinine.Ultrasound indicated a significantly elevated flow rate of >300 cm/s( n=4)and CTA/MRA indicated that the degrees of stenosis varied from 50% to 95%( n=5). Results:After balloon dilatation, stenosis either improved( n=2)or relapsed( n=2). Further stenting succeed( n=1)or failed( n=1). One case of stenosis was relieved partially after 6-month observation without any invasive treatment. Conclusions:As a serious complication, transplant renal artery stenosis is common after pediatric donor kidney transplantation.Too small size in donor kidney and rapid recipient growth may be specific risk factors.After diagnosis, balloon dilation is a preferred treatment.Stent placement should be cautiously employed.

Early Diagnosis and Clinical Significance of Acute Cardiac Injury - Under the Iceberg: A Retrospective Cohort Study of 619 Non-critically Ill Hospitalized COVID-19 Pneumonia Patients

RationaleCoronavirus disease 2019 (COVID-19) can cause a viral pneumonia together with other extrapulmonary complications. Acute cardiac related injury (ACRI) is common in hospitalized COVID-19 patients. ObjectiveTo explain the pathological mechanism of ACRI and improve the treatment strategy by retrospectively observing the factors associated with ACRI and factors affecting the prognosis of ACRI with COVID-19 at an early stage. Methods619 COVID-19 patients were from Tongji Hospital, Wuhan. Students t test was used for continuous variables while Pearson {chi}2 test for categorical factors. Univariable and multivariable logistic regression models were applied to estimate odds ratio (OR) with 95% confidence interval (CI). ResultsAmong the 619 OOS Level-I hospitalized COVID-19 patients, 102 (16.5%) were defined as ACRI (stage-1: 59 cases, stage-2: 43 cases). 50% of ACRI patients developed into severe cases and 25 patients died(CFR=24.5%), 42 times that of non-ACRI patients. Elderly (OR=2.83, P<0.001), HTN (OR=2.09, P=0.005), {gamma}-globulin (OR=2.08, P=0.004), TCM (OR=0.55, P=0.017), PLT (OR=2.94, P<0.001) and NLR (OR=2.20, P=0.004) were independently correlated with ACRI. SBP [>=] 140, dyspnea, DM, smoking history were correlated with ACRI-stage2 only. In the prognostic subgroup analysis of ACRI patients, {gamma}-globulin treatment could prolong LOS (29.0 {+/-} 7.2 days Vs 23.5 {+/-} 8.1 days, P=0.004). TCM (OR=0.26, P=0.006), SBP [>=] 160 (OR= 22.70, P=0.005), male (OR=2.66, P=0.044) were associated with severe illness while corticosteroids treatment (OR=3.34, P=0.033) and male (OR=4.303, P=0.008) with death. Surprisingly, we found the mortality of non-elderly patients is higher than elderly (32.4% VS 20.0%, P=0.164), and both IKF and RASI treatment were not correlated with any prognostic indicators including severe, death and LOS. ConclusionThis study observed that several non-traditional issues were associated with early cardiac injury in COVID-19 while many traditional cardiovascular risk factors were not. Besides elderly and male, hypertension was confirmed to be the most important risk factor.

Impact of Chronic Comorbidities on Progression and Prognosis in Patients with COVID-19: A Retrospective Cohort Study in 1031 Hospitalized Cases in Wuhan, China

ObjectivesThe main aim of our study is to observe clinical characteristics and effects of antihypertensive drugs in different hospitalized populations, and to further provide evidence to explore causes and specific clinical markers of the aggravation of COVID-19 in patients with underlying hypertension. DesignThis was a retrospective cohort study focusing on the clinical data of COVID-19 inpatients admitted at the early stage of pandemic. SettingA single center study conducted in Tongji Hospital, Tongji Medical College of Huazhong university of Science and Technology (Wuhan, China). ParticipantsAll 1031 inpatients diagnosed with COVID-19 according to Prevention and control Scheme for Novel Coronavirus Pneumonia published by National Health Commission of the Peoples Republic of China and WHO interim guidance in Tongji hospital (Wuhan, China), from January 27, 2020, to March 8, 2020 with the cutoff date being March 30, 2020, were included in this study. Main outcome measuresDemographic data, medical history, clinical symptoms and signs, laboratory findings, chest computed tomography (CT), treatment, and clinical outcomes were extracted from electronic medical records. Results1031 COVID-19 inpatients were included in this study, of whom 866 were discharged and 165 were deceased in hospital. 73% of 165 deceased patients had chronic comorbidities. Patients with underlying diseases showed CFR 2.8 times as that of patients without. Senility and males were observed to be main risk factors for increased in-hospital case-fatality rate, with the odds ratio in multivariable regression being 2.94 (95%CI: 2.00 to 4.33; P <0.001) and 2.47 (95%CI: 1.65 to 3.70; P <0.001), respectively. The odds ratio of cases with composite endpoints for patients with simple hypertension was 1.53 (95%CI: 1.07 to 2.17; P=0.019). Senile patients with hypertension were proved to be at high risk early in the disease, which might be associated with the level of CRP, LDH, and eGFR. The odds ratio of case-fatality rate for patients with hypertension taking CCB group was 0.67 (95%CI: 0.37 to 1.20; P = 0.176). Among 271 severe cases without IKF, the odds ratio of case-fatality rate was 0.42 (95CI%: 0.18 to 0.99; P = 0.046) for patients in the CCB group after adjustment of age, sex, and underlying diseases. ConclusionsHypertension is not just a chronic underlying comorbidity, but also a risk factor affecting the severity of COVID-19 and does play a critical role in worsening patients clinical outcomes. Therefore, hypertension management in patients with COVID-19 should be regarded as a major challenge in the diagnostic and therapeutic strategies. Trial registrationN.A.

Derivation and validation of a prognostic model for predicting in-hospital mortality in patients admitted with COVID-19 in Wuhan, China: the PLANS (Platelet Lymphocyte Age Neutrophil Sex) model

OBJECTIVETo develop and validate a prognostic model for in-hospital mortality in COVID-19 patients using routinely collected demographic and clinical characteristics. DESIGNMulticenter, retrospective cohort study. SETTINGJinyintan Hospital, Union Hospital, and Tongji Hosptial in Wuhan, China. PARTICIPANTSA pooled derivation cohort of 1008 COVID-19 patients from Jinyintan Hospital, Union Hospital in Wuhan and an external validation cohort of 1031 patients from Tongji Hospital in Wuhan, China. MAIN OUTCOME MEASURESOutcome of interest was in-hospital mortality, treating discharged alive from hospital as the competing event. Fine-Gray models, using backward elimination for inclusion of predictor variables and allowing non-linear effects of continuous variables, were used to derive a prognostic model for predicting in-hospital mortality among COVID-19 patients. Internal validation was implemented to check model overfitting using bootstrap approach. External validation to a separate hospital was implemented to evaluate the generalizability of the model. RESULTSThe derivation cohort was a case-mix of mild-to-severe hospitalized COVID-19 patients (n=1008, 43.6% females, median age 55). The final model (PLANS), including five predictor variables of platelet count, lymphocyte count, age, neutrophil count, and sex, had an excellent predictive performance (optimism-adjusted C-index: 0.85, 95% CI: 0.83 to 0.87; averaged calibration slope: 0.95, 95% CI: 0.82 to 1.08). Internal validation showed little overfitting. External validation using an independent cohort (n=1031, 47.8% female, median age 63) demonstrated excellent predictive performance (C-index: 0.87, 95% CI: 0.85 to 0.89; calibration slope: 1.02, 95% CI: 0.92 to 1.12). The averaged predicted survival curves were close to the observed survival curves across patients with different risk profiles. CONCLUSIONSThe PLANS model based on the five routinely collected demographic and clinical characteristics (platelet count, lymphocyte count, age, neutrophil count, and sex) showed excellent discriminative and calibration accuracy in predicting in-hospital mortality in COVID-19 patients. This prognostic model would assist clinicians in better triaging patients and allocating healthcare resources to reduce COVID-19 fatality.

CYP2J2 and its metabolites (EETs) attenuate cardiac hypertrophy by activating AMPKα2 and enhancing nuclear translocation of Akt1 / 中国病理生理杂志

AIM:Cytochrome P450 epoxygenase 2J2 and epoxyeicosatrienoic acids ( EETs) are known to protect against cardiac hypertrophy and heart failure, which involve activation of 5′-AMP-activated protein kinase ( AMPK) and Akt.Although the functional roles of AMPK and Akt are well established , the significance of crosstalk between them in the development of cardiac hypertrophy and anti -hy-pertrophy of CYP2J2 and EETs remains unclear .Here, we investigated whether CYP 2J2 and its metabolites EETs protected against cardiac hypertrophy by activating AMPKα2 and Akt1.Moreover, we tested whether EETs enhanced crosstalk between AMPKα2 and phosphorylated Akt1 ( p-Akt1), and stimulated the nuclear translocation of p-Akt1, to exert their anti-hypertrophic effects. METHODS:The recombinant rAAV9 vector was coupled to CYP2J2 and the rAAV9-CYP2J2 construct was injected into the caudal vein of AMPKα2-/-and littermate control mice .AMPKα2 -/-and littermate control mice that overexpressed CYP 2J2 in heart were treated with angiotensin II (Ang II) for 2 weeks.Hemodynamic and cardiac functions were also evaluated after 14 days of infusion with Ang II or saline.RESULTS:Interestingly, the overexpression of CYP2J2 suppressed cardiac hypertrophy , including decreased heart size, cross sectional area of cardiomyocytes , markers of cardiac hypertrophy [ brain natriuretic peptide ( BNP) ,β-myosin heavy chain (β-MHC) and skeletal muscle α-actin (ACTA1)] and increased levels of atrial natriuretic peptide (ANP) in the heart tissue and plasma of wild-type mice but not AMPKα2 -/-mice.Measurement of left ventricular ejection fraction and fractional shortening showed that CYP2J2 overexpression prevented Ang II-induced ventricular systolic dysfunction in mice .Moreover, an Ang II-induced reduction in cardiac function, demonstrated by decreased dp/dtmax and dp/dtmin, was prevented by overexpression of CYP2J2.Mechanistically, the CYP2J2 metabolites 11,12-EET activated AMPKα2 to induce the nuclear translocation of p-Akt1, which increased production of ANP and thereby inhibited the development of cardiac hypertrophy .Furthermore , by co-immunoprecipitation analysis , we found that full-length Akt1 and an Akt1 fragment containing amino acids 150-408, which constitute the protein kinase domain , but not other frag-ments of Akt1, bind to the AMPKγ1 subunit.AMPKα2β2γ1 and p-Akt1 interact through the direct binding of the AMPKγ1 subunit to the Akt1 protein kinase domain.This interaction was enhanced by 11,12-EET.CONCLUSION:Our studies reveal a novel mechanism in which CYP2J2 and EETs enhanced Akt1 nuclear translocation through interaction with AMPKα2β2γ1 and protect against cardiac hy-pertrophy and suggest that overexpression of CYP 2J2 might have clinical potential to suppress cardiac hypertrophy and heart failure .

Relationship between microalbuminuria to estimated glomerular filtration rate ratio and incidence of contrast induced nephropathy / 实用医学杂志

Objective To investigate the relationship between microalbuminuria to estimated glomerular filtration rate (mALB/GFR) ratio and incidence of contrast induced nephropathy (CIN) after underwent percutaneous coronary intervention (PCI). Between March 2010 to August 2011, a total of 137 patients who underwent PCI were selected. Anthropometric measures, lipid profiles, fasting blood glucose, CRP, base creatinine and microalbuminuria were measured before operation. Glomerular filtration rate was calculated by MDRD formula. Contrast volume (CV) was recorded after the operation for each patient. Serum creatinine was measured 24 h, 48 h, 72 h after operation, respectively. CIN was defined as an increase in serum creatinine levels of 25% or 44 μmol/L or more from baseline within 72 h after PCI. All patients were divided into group A (CIN group) and group B (No CIN group). Results Eighteen (13.1%) patients developed CIN(group A). The others were group B (no CIN group). The level of CRP, base creatinine, microalbuminuria and mALB/GFR in group A were significantly higher than that in group B ( P < 0 . 01 ) . GFR in group A was lower than that in group B (P < 0.05). Logistic regression analysis indicated that the level of base creatinine , microalbuminuria GFR and mALB/GFR were independent risk factors of CIN occurrence (P < 0.05). The receiver-operator characteristic curve analysis indicated that a mALB/ GFR ratio of 1.17 was a fair discriminator for CIN,and sensitivity were 94.1%, specifity were 72.5%. Conclusion A mALB/ GFR ratio was a independent predictor of CIN after PCI.

Nine-month angiographic and two-year clinical follow-up of polymer-free sirolimus-eluting stent versus durable-polymer sirolimus-eluting stent for coronary artery disease: the Nano randomized trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>First generation drug-eluting stents (DES) were associated with a high incidence of late stent thrombosis (ST), mainly due to delayed healing and re-endothelization by the durable polymer coating. This study sought to assess the safety and efficacy of the Nano polymer-free sirolimus-eluting stent (SES) in the treatment of patients with de novo coronary artery lesions.</p><p><b>METHODS</b>The Nano trial is the first randomized trial designed to compare the safety and efficacy of the Nano polymer-free SES and Partner durable-polymer SES (Lepu Medical Technology, Beijing, China) in the treatment of patients with de novo native coronary lesions. The primary endpoint was in-stent late lumen loss (LLL) at 9-month follow-up. The secondary endpoint was major adverse cardiac events (MACE), a composite of cardiac death, myocardial infarction or target lesion revascularization.</p><p><b>RESULTS</b>A total of 291 patients (Nano group: n = 143, Partner group: n = 148) were enrolled in this trial from 19 Chinese centers. The Nano polymer-free SES was non-inferior to the Partner durable-polymer DES at the primary endpoint of 9 months (P < 0.001). The 9-month in-segment LLL of the polymer-free Nano SES was comparable to the Partner SES (0.34 ± 0.42) mm vs. (0.30 ± 0.48) mm, P = 0.21). The incidence of MACE in the Nano group were 7.6% compared to the Partner group of 5.9% (P = 0.75) at 2 years follow-up. The frequency of cardiac death and stent thrombosis was low for both Nano and Partner SES (0.8% vs. 0.7%, 0.8% vs. 1.5%, both P = 1.00).</p><p><b>CONCLUSIONS</b>In this multicenter randomized Nano trial, the Nano polymer-free SES showed similar safety and efficacy compared with the Partner SES in the treatment of patients with de novo coronary artery lesions. Trials in patients with complex lesions and longer term follow-up are necessary to confirm the clinical performance of this novel Nano polymer-free SES.</p>

Experimental study of CD151 gene therapy on improving myocardial function in swines with myocardial infarction / 中华老年医学杂志

Objective To investigate the effect of CD151 gene therapy on improving myocardial function in swines with myocardial infarction. Methods CD151, antisense CD151 and green fluorescent protein (GFP) were constructed into the recombinant adeno-associated virus (rAAV). Swines were divided into 4 groups: rAAV-GFP group (6 swines), rAAV-CD151 group (6 swines), rAAV-antiCD151 group (6 swines) and control group (6 swines). The swines were performed with coronary artery ligation and intramuscularly injection with rAAV. Eight weeks after vector administration, western blot was used to detect gene expression of CD151. 13N-labeled NH3 positron emission tomography (PET) was used to evaluate myocardial perfusion. Echocardiography was used to assess myocardial function. Results Compared with the control group and the rAAV-GFP group, the rAAV-CD151 group showed higher CD151 protein expression. Compared with the rAAV-GFP group, the defect size of myocardium was decreased[( 11.3±2.4)% vs. (21.1±2.6)%, t= -5.67,P<0.01] and left ventricular ejection fraction (EF), left ventricular fractional shortening (FS), the ratio of anterior lateral wall thickening (△ALWT) and ratio of interventricular septum thickening (△IVST) were significantly improved in rAAV-CD151 group 8 weeks after vector administration [(65.7±4.6)% vs. (54.7±5.3)%, (36.0±2.9)% vs. (27.6±3.1)%,(55.4± 4.9)% vs. (36.8±7.8)%, (35.2±6.0)% vs. (26.7±4.4)%, t=3.98, 3.35, 3.34, 9.27, all P< 0.05]. The level of diastolic ALWT and diastolic IVST was also increased in rAAV CD151 group compared with rAAV-GFP group ( P<0.05).Compared with rAAV-CD151 group, parameters of myocardial function in rAAV-antisense CD151 group were not improved (P<0.05). Conclusions rAAV-CD151 can effectively transfeet the myocardium, increase the expression ofCD151 protein, promote the blood perfusion of myocardium and improve the ventricular function after myocardial infarction.

Observation on cerebral microbleeds in patients with hypertension by magnetic resonance imaging / 中华神经科杂志

Objective To concentrate on the morbidity of cerebral microbleed (CMB) in patients with hypertension and to analyze the predilection and risk-factor of cerebral microbleed.Method Hypertensive patients were divided into the simple hypertention group, hypertention group with lacunar infarction and hypertention group with cerebral infarction.All of these 65 patients received examination of susceptibility-weighted imaging.Results Ninety-one focuses of cerebral microbleeds were found in these patients:58.2% of these focuses were in both basal ganglia and cerebral ganglion;35.2 percent were in cortex and subcortex;6.6 percent were in brainstem and cerebellum.The total morbidity of CMB was 33.8 percent, 52.4 percent in the group with lacunar infarction and 38.1 percent in the group with cerebral infarction, both were significantly higher than that of 8.7 percent in the simple hypertensive group (χ2= 8.08,P<0.01 andχ2=3.86, P<0.05).Conclusions The focus of CMB suggested the hemorrhagic tendency in endocranial capillary.CMB can be used as a routine exam for the hemorrhagic tendency in endocranial capillary.Synthetic analysis of risk-factor and the result of SWI help clinicians choose suitable treatment for each patient.

Role of leptin in atherogenesis.

The pathogenesis of atherosclerosis involves multiple cellular events, including endothelial cell dysfunction, inflammation, proliferation of vascular smooth muscle cells and matrix alteration that is subsequently characterized by hardening, thickening, loss of elasticity and, finally, a reduction in the vessel's lumen. Leptin, a peptide hormone, is produced by adipocytes, and the majority of obese individuals have high plasma leptin concentrations. Leptin regulates food intake as well as metabolic function. Originally thought to be a satiety factor, leptin is a pleiotropic molecule. In addition to its metabolic effects, leptin regulates the production of several pro- and anti-inflammatory cytokines by activating immune cells. It is associated with increased plasma C-reactive protein concentrations, vascular proliferation, calcification and decreased arterial distensibility. Leptin also increases oxidative stress. Moreover, leptin contributes to increases in blood pressure, and thus, probably plays an important role in the initiation and progression of atherosclerosis. 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) lower lipid concentrations and also decrease endothelial apoptosis, inhibit smooth muscle cell proliferation, and lower concentrations of C-reactive protein and proinflammatory cytokines; moreover, it is now known that statins can inhibit leptin release by adipocytes. Therefore, statins have been shown to be beneficial in atherosclerosis. The present review mainly focuses on the various evidence that suggest a potential atherogenic mechanism of leptin, and also briefly addresses the beneficial role of statins in atherosclerosis.

Effects of Simvastatin on NF-κB-DNA Binding Activity and Monocyte Chemoattractant Protein-1 Expression in a Rabbit Model of Atherosclerosis / 华中科技大学学报（医学）（英德文版）

To observe the effects of simvastatin on nuclear factor kappaB (NF-κB)-DNA binding activity and on the expression of monocyte chemoattractant protein-1 (MCP-1) in atherosclerotic plaque in rabbits and to explore the anti-atherosclerotic properties beyond its lipid-lowering effects.Thirty-six New Zealand male rabbits were randomly divided into low-cholesterol group (LC), highcholesterol group (HC), high-cholesterol+ simvastatin group (HC+S) and then were fed for 12weeks. At the end of theexperiment, standard enzymatic assays, electrophoretic mobility shift assay (EMSA), immunohistochemical staining, and morphometry were performed to observe serum lipids, NF-κB-DNA binding activity, MCP-1 protein expression, intima thickness and plaque area of aorta respectively in all three groups. Our results showed that the serum lipids, NF-κB-DNA binding activity, expression of MCP-1 protein, intima thickness, and plaque area of aorta in the LC and HC+S groups were significantly lower than those in the HC group (P＜0.05). There was no significant difference in the serum lipids between the LC and HC+S groups (P＞0.05), but the NF-κB-DNA binding activity, the expression of MCP-1 protein and the intima thickness and plaque area of aorta in the HC+S group were significantly decreased as compared to the LC group (P＜0.05). This study demonstrated that simvastatin could decrease atherosclerosis by inhibiting the NFκB-DNA binding activity and by reducing the expression of MCP-1 protein.

Effects of leptin on the expression of tumor necrosis factor-alpha in RAW264.7 cells / 中国组织工程研究

BACKGROUND: Leptin is a kind of polypeptide hormone secreted by fatty tissue, previous studies have shown that leptin plays a certain role in the formation of atherosclerosis.OBJECTIVE: To investigate the effects of leptin on the expression of tumor necrosis factor-alpha (TNF-α) in RAW264.7 cells, and investigate the possible mechanism from the angle of the change of nuclear factor-κB (NF-κB) activity.DESIGN: A controlled observational experiment.SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiments were carried out in the Department of Biochemistry and Molecular Biology, Tongji Medical College, and the Department of General Surgery, Union Hospital affiliated to Huazhong University of Science and Technology between April 2005 and February 2006.The cultured RAW264.7 cells were divided into leptin treated groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. Each group had 3 sub-wells,and the experiments were repeated for 3 times.METHODS: Mouse macrophage RAW264.7 cells were incubated into a 6-well plate at the density of 109 cells L-1, cultured in RPMI-1640 culture medium containing bovine serum of 0.1 in volume fraction. When the cells grew to 80%, serum-free culture medium Opti-MEM was changed to culture for another 24 hours, and then the cells were divided into leptin groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. After the cells were incubated with leptin for 4 hours, the expression of TNF-α mRNA expression in RAW264.7 cells was detected with reverse transcription-polymerase chain reaction (RT-PCR). After the RAW264.7 cells were incubated with leptin for 1, 3, 6 and 9 hours, the expression of TNF-c protein expression was detected with double antibody sandwich enzyme-linked immunosorbent assay (ELISA). After the RAW264.7 cells were incubated with leptin for different times, the activity of NF-κB was detected analyzed with electrophoretic mobility shift assay. Another, the RAW264.7 cells were treated with or without 50 μmol/L leptin and/or 100 μmol/L PS1145 (I kappa B kinase specific inhibitor)divided into four groups: blank control group, I kappa B kinase specific inhibitor PS1145 (10 μmol/L) treated group, leptin (50 μmol/L) treated group, leptin (50 μmol/L) + PS1145 (10 μmol/L) group. Aftere incubated for 6 hours, the activity of NF-κB and expression TNF-α mRNA were detected respectively.MAIN OUTCOME MEASURES: ① Effect of leptin of different concentration on the expression of TNF-α mRNA and protein in RAW264.7cells; ② Effect of leptin of different concentration on the activity of NF-κB in RAW264.7 cells; ③ Influence of inhibition I kappa B kinase activity inhibition on expression of TNF-a induced by leptin in RAW264.7cells.RESULTS: ① After the RAW264.7 cells were treated with leptin of different concentration, the TNF-α mRNA level was elevated in a dose-dependent manner, and it reached the peak value emerged in the 50 μg/L leptin treated group. ② The expression of TNF-α protein increased in dose-dependent and time-dependent manners, and it reached the peak val ue at 6 hours in the 50 μg/L leptin treated group. ③ The activity of NF-κB was also positively correlated with the leptin concentration, and it was the highest value at 6 hours treated with 50 μg/L leptin (P ＜ 0.05). ④ I kappa B kinase activity inhibition only partially suppressed the leptin induced elevation of TNF-α expression induced by leptin.CONCLUSION: Leptin can increase the expression and secretion of TNF-α in RAW264.7 cells directly in both dose-dependent and time-dependent manners, and the mechanism may be correlated with the activated NF-κB induced by leptin. It may be one of the mechanisms of atherosclerosis induced by leptin.

Effects of simvastatin on NF-kappaB-DNA binding activity and monocyte chemoattractant protein-1 expression in a rabbit model of atherosclerosis / 华中科技大学学报（医学）（英德文版）

To observe the effects of simvastatin on nuclear factor kappaB (NF-kappaB)-DNA binding activity and on the expression of monocyte chemoattractant protein-1 (MCP-1) in atherosclerotic plaque in rabbits and to explore the anti-atherosclerotic properties beyond its lipid-lowering effects. Thirty-six New Zealand male rabbits were randomly divided into low-cholesterol group (LC), high-cholesterol group (HC), high-cholesterol+simvastatin group (HC+S) and then were fed for 12 weeks. At the end of the experiment, standard enzymatic assays, electrophoretic mobility shiftassay (EMSA), immunohistochemical staining, and morphometry were performed to observe serum lipids, NF-kappaB-DNA binding activity, MCP-1 protein expression, intima thickness and plaque area of aorta respectively in all three groups. Our results showed that the serum lipids, NF-kappaB-DNA binding activity, expression of MCP-1 protein, intima thickness, and plaque area of aorta in the LC and HC+S groups were significantly lower than those in the HC group (P0.05), but the NF-kappaB-DNA binding activity, the expression of MCP-1 protein and the intima thickness and plaque area of aorta in the HC+S group were significantly decreased as compared to the LC group (P<0.05). This study demonstrated that simvastatin could decrease atherosclerosis by inhibiting the NF-kappaB-DNA binding activity and by reducing the expression of MCP-1 protein.

Danshen inhibits oxysterol-induced endothelial cell apoptosis in vivo.

Oxysterols induce apoptosis in vascular endothelial cells in vitro, but it is not clear whether they do so in vivo. We intravenously injected an oxysterol, cholestan-3beta, 5alpha, 6beta-triol, into rats and quantitatively analyzed endothelial cell apoptosis in the aorta. Oxysterol significantly promoted apoptosis in a time- and dose-dependent fashion. The apoptosis had increased 4.5-fold 6 hrs after injection, and returned to the background level at 48 hrs. An extract of the Chinese herb Danshen as well as probucol abolished triol-induced endothelial cell apoptosis in vitro and in vivo. Since apoptotic cells are quickly cleared, oxysterol-induced apoptosis could significantly affect endothelial integrity over a long period of time. Radical scavengers may be useful for the prevention of endothelial damage.

Effects of losartan and ginseng on cardiomyocyte Bcl-2 gene expression induced by ischemia and reperfusion / 中国组织工程研究

BACKGROUND: The studies found that cardiomyocyte apoptosis is related to ischemia-reperfusion injury directly. The clinical and experimental studies proved that ginseng and losartan could improve myocardial iscbemia and prevent the ischemia-reperfusion injury.But the comparative study of their effects on cardiomyocyte injury induced by ischemia and reperfusion has not been reported.OBJECTIVE: To compare the effects of ginseng and losartan on cardiomyocyte Bcl-2 gene expression after ischemia and reperfusion in vivo.DESIGN: Randomized controlled experiment.SETTING: Department of Cardiovascular Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Laboratory of Cardiovascular Internal Medicine of Tongji Hospital of Tongji Medical College Affiliated to Huazhong University of Science and Technology from November 2002 to April 2003. Totally 40 healthy adult Wistar rats, weighting 200-250 g, of either gender, were selected and divided into 4 groups:sham operated group, ischemia-reperfusion group, ginseng treated group and losartan treated group with 10 rats in each group.METHODS: Rats were modeled in ischemia-reperfusion group, ginseng treated group and los arran treated group but not in sham operation group.20 mg/kg (1 mL in volume) losartan was given by stomach. The first administration was 2 hours prior to operation. Subsequently, the second and third administrations were given in immediate and 24 hours after operation,respectively; 1 mL of radix ginseng rubra (1 g/mL) was given by stomach.The first administration was 2 hours before operation. Subsequently, the second and third administrations were given in just and 24 hours after operation, respectively. The rats in ischemia-reperfusion and sham-operated group were given the normal saline with the same volume and at the same time. Immunohistochemistry and in situ hybridization histochemistry were used to measure mRNA and protein of Bcl-2 gene expression that were compared with those in the control group.MAIN OUTCOME MEASURES: The expression level of Bcl-2 mRNA and Bcl-2 protein in ginseng treated group and losartan treated group.RESULTS: Data of totally 40 rats was entered the final analysis. ① Content of Bcl-2 mRNA and protein were not significantly different both in ginseng group and control group (P ＞ 0.05). ② Contents of Bcl-2 mRNA and Bcl-2 protein were higher in ginseng treated group than those in the ischemia-reperfusion group (P ＜ 0.05 or 0.01).CONCLUSION: Expressions of Bcl-2 mRNA and Bcl-2 protein in ginseng treated group are higher than those in losartan treated group, which suggests that ginseng has not same effect with losartan in inhibiting cardiomyocyte apoptosis and in preventing and curing cardiomyocyte injury induced by ischemia-reperfusion.

Effects of carvedilol on cardiomyocyte apoptosis and gene expression in vivo after ischemia-reperfusion in rats / 华中科技大学学报（医学）（英德文版）

The effects of carvedilol on cardiomyocyte apoptosis and expression of bcl-2, bax genes following ischemia (0.5 h) and reperfusion (48 h) in vivo and the possible biological mechanism of carvedilol inhibiting cardiomyocyte apoptosis were studied. The left anterior descending artery in Wistar rats were ligated to establish ischemia-reperfusion (I/R) models. The model animals were divided into two groups: I/R group, the model rats not subject to other treatments except ischemia-reperfusion (n = 8); carvedilol-treated group (n = 8), I/R model rats treated with carvedilol. Eight rats in the sham-operated group were subjected to only experimental open operation. The number of apoptotic cardiomyocyte was determined by TUNEL staining. Immunohistochemistry and in situ hybridization histochemistry (ISHH) were used to detect the expression of bcl-2 and bax genes. Image processing system was used to quantitatively dispose the positive metric substances of both immunohistochemistry and ISHH through the average optical density (OD) value. The results showed that the number of the apoptotic cells were 36.18 +/- 9 (I/R group), 0-1 (sham-operated group) and 9.5 +/- 3 (carvedilol-treated group) in each visual field respectively with the difference being very significant among the groups (P < 0.001). The OD values of bcl-2 protein in sham-operated group, I/R group and carvedilol-treated group were 0.14 +/- 0.01, 0.08 +/- 0.02 and 0.15 +/- 0.03, respectively. The OD values of bcl-2 mRNA in sham-operated group, I/R group and carvedilol-treated group were 0.08 +/- 0.01, 0.06 +/- 0.01 and 0.09 +/- 0.01, respectively. There was no significant difference between carvedilol-treated group and I/R group (P > 0.05). The OD values of bax protein in I/R group, sham-operated and carvedilol-treated-treated group were 0.13 +/- 0.02, 0.07 +/- 0.01, 0.06 +/- 0.01, respectively. There was very significant difference between carvedilol-treated group and I/R group (P < 0.01). Bcl-2/bax ratio was 1.07 +/- 0.14 (I/R group), 1.28 +/- 0.16 (sham-operated group), 2.5 +/- 0.26 (carvedilol-treated group) respectively with the difference being very significant between carvedilol-treated group and I/R group (P < 0.05). It was indicated that carvedilol could inhibit cardiomyocyte apoptosis following ischemia and reperfusion, which was related to the increased bcl-2/bax ratio due to inhibition of bax gene expression.

Effect of ginsenoside-Rb_1 and Re against cardiomyocyte apoptosis and expression of the related gene proteins in the experimental cardiac ischemia-reperfusion in rats / 中华物理医学与康复杂志

Objective To observe the effect of ginsenoside Rb 1 and Re on cardiomyocyte apoptosis and expression of Bcl-2, Bax, Bad and Fas gene proteins after acute ischemia and reperfusion in rats and elucidate the possible mechanisms. Methods The ischemia-reperfusion heart model was made by ligating the left anterior descending branch of coronary artery in Wistar rats. The apoptotic cardiomyocytes were confirmed with transmission electron microscopy and counted with in situ nick labeling (TUNEL) method and light microscopy. The expression of Bcl-2?Bax?Bad and Fas gene proteins were studied by immunohistochemical staining. Mean optical density (OD) value of the gene proteins expression were quantitatively examined by image analysis system. Results A.The apoptotic cardiomyocytes were not observed in the sham-operation group. The number of the apoptotic cells were 134.45?45.61/field in the ischemia- reperfusion group, 51.65?13.71/field in the ginsenoside Rb 1-treated group and 90.66? 19.22/field in the ginsenoside Re-treated group, respectively. The differences were significant among the three groups (P0.05), but Bax?Bad and Fas gene expression were decreased significantly in the ginsenoside Rb 1 and Re-treated group as compared with the ischemia-reperfusion group (P