The purpose of this study was to explore the investigative mechanism of salidroside (SAL) on LPS-induced acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). The exosomes from RLE-6TN are extracted and identified by transmission electron microscopy, particle size analysis and protein marker detection, and co-cultured with NR8383 cells. The ALI/ARDS model of SD rats was established by LPS (10 mg/kg) intratracheal instillation. Following a four-hour intratracheal instillation of LPS, 50 μl of RLE-6TN exosomes were injected through the tail vein. After that, SAL and miR-146a antagomir were injected into the tail vein for 72 h, respectively. As the changes of HE stain, body weight and ALI score are observed. The expression of miR-146a, TLR4, NF-kB, IRAK1, TRAF6 and their related proteins were detected by RT-PCR and Western blot, respectively. TNF-α, IL-6, IL-8 and IL-1 β inflammatory factors were detected by ELISA. The expression of miR-146a, NF-kB, IRAK, TRAF6 and related inflammatory factors in LPS-induced NR8383 was significantly higher than that in the control group, while SAL has greatly reduced the expression of TLR4 mediated NF-kB inflammatory pathway and related inflammatory factors. SAL can significantly improve the LPS-induced lung morphological abnormalities, slowed down the rate of weight loss in rats, and reducing the ALI score. The expression trend of NF-kB, IRAK, TRAF6 and related inflammatory factors in rats’ lung tissues was consistent with that in NR8383 cells. SAL has a protective effect on ALI/ARDS caused by sepsis, which is likely to be developed to a potential treatment for the disease. To sum up, this study provides a new theoretical basis for the treatment of ALI/ARDS with SAL.

Thymosin-beta 4 (T beta 4) has been reported to exert a pro-angogenic effect on endothelial cells. However, little is known on the role and underlying mechanisms of T beta 4 on critical limb ischemia (CLI). The present study aimed there-fore to investigate the mechanisms and pro-angiogenic effects of T beta 4 in CLI mice. T beta 4 overexpression lentiviral vector was first transfected into HUVEC and CLI mice model, and inhibitors of Notch pathway (DAPT) and NF-kappa B pathway (BMS) were also applied to HUVEC and CLI mice. Subsequently, MTT, tube formation and wound healing assays were used to determine the cell viability, angiogenesis and migratory ablity of HUVEC, respectively. Western blotting, reverse transcription, quantitative PCR, immunofluorescence and immunohistochemistry were used to detect the expression of the angiogenesis-related factors angiopoietin-2 (Ang2), TEK receptor tyrosine kinase 2 (tie2), vascular endothelial growth factor A (VEGFA), CD31 and alpha-smooth muscle actin (alpha-SMA) and the Notch/NF-kappa B pathways-related factors NOTCH1 intracellular domain (N1ICD), Notch receptor 3 (Notch3), NF-kappa B and p65 in HUVEC or CLI mice muscle tissues. The results demonstrated that T beta 4 not only enhanced the cell viability, angiogenesis and migratory ability of HUVEC but also promoted the expression of Ang2, tie2, VEGFA, N1ICD, Notch3, NF-kappa B, and phosphorylated (p)-p65 in HUVEC. In addition, T beta 4 promoted the expression of CD31, alpha-SMA Ang2, tie2, VEGFA, N1ICD and p-p65 in CLI mice muscle tissues. Treatment with DAPT and BMS had opposite effects of T beta 4, whereas T beta 4 reversed the effect of DAPT and BMS. The findings from the present study suggested that T beta 4 may promote angiogenesis in CLI mice via regulation of Notch/NF-kappa B pathways.

OBJECTIVE:To study whether tricalcium phosphate(TCP) wear particles cause injuries of periprosthetic osteocytes in the mouse calvaria, and to explain its molecular mechanism.METHODS:Thirty six-week(ICR)male mice were randomly divided into sham group, model (TCP) group and 3-methyladenine (3-MA) group. A murine calvarial model of osteolysis was established by 30 mg of TCP wear particles implantation over the periosteum around the middle suture of calvaria in mice. On the second postoperative day, the autophagy specific inhibitor 3-MA (1.0 mg/kg) was subcutaneously injected to the calvaria in the 3-MA-treated mice every other day. After 2 weeks, blood and the calvaria were obtained. Micro-CT was used to detect bone mineral density(BMD), bone volume fraction (BVF) and porosity number. HE staining and flow cytometry were performed to analyze the viability and apoptosis of periprosthetic osteocytes. The serum levels of dentin matrix protein 1(DMP-1) and sclerostin (SOST) were determined by ELISA. The proteins expressions of DMP-1, SOST, Beclin-1 and microtuble-associated protein 1 light chain 3 (LC-3) were detected by Western blot in the calvaria osteocytes.RESULTS:Compared with the sham group, the mice in the TCP group showed that a significant decrease in the viability of periprosthetic osteocytes, but obvious increases in number of osteocytes death and osteocytes apoptosis (P<0.05), and in serum level and protein expression of SOST; significant decreases in serum level and protein expression of DMP-1 (P<0.05), and remarkable up-regulation of autophagy-related factors beclin-1 and the conversion of LC3-Ⅱ from LC3-I in the calvaria osteocytes. Compared with TCP group, the mice in the 3-MA group showed that injuries of calvaria osteocytes were obviously aggravated, and osteocytes apoptosis was significantly increased (P<0.05).CONCLUSIONS:TCP wear particles can cause injuries of periprosthetic osteocytes via activation of apoptosis and autophagy, which promotes osteolysis around the prosthesis osteolysis and joint aseptic loosening.

Wear debris-induced osteolysis and aseptic loosening are the most frequent late complications of total joint arthroplasty leading to revision of the prosthesis. However, no effective measures for the prevention and treatment of particles-induced osteolysis currently exist. Here, we investigated the efficacy of local administration of osthole on tricalcium phosphate (TCP) particles-induced osteolysis in a murine calvarial model.

ObjectiveTo investigate the relationships between traditional Chinese medicine(TCM) syndrome differentiation and serum cystatin C(Cys-C) and homocysteine(Hcy) in patients with chronic heart failure(CHF). Methods 115 cases with CHF admitted into the Department of Cardiology of the First Affiliated Hospital of Zhejiang Chinese Medical University were selected in the CHF group, and 30 cases who had taken health examination in the same period were chosen in the healthy control group. According to the TCM syndrome differentiation, the CHF cases were subdivided into four groups with different types of syndrome: 30 cases of deficiency of both Qi and Yin syndrome, 30 cases of Qi deficiency syndrome and blood stagnation syndrome, 30 cases of heart and kidney Yang deficiency syndrome and 25 casesof flooding due to Yang deficiency syndrome. The serum levels of Cys-C and Hcy in different groups were tested, and the relationships between TCM syndrome differentiation and serum Cys-C and Hcy were analyzed by using Spearman rank correlation analysis.Results The serum levels of Cys-C and Hcy in the patients with CHF were significantly higher than those in the healthy control group〔Cys-C(mg/L):1.24±0.34 vs. 0.77±0.22, Hcy(μmol/L):18.66±4.57 vs. 11.65±3.21,bothP<0.05〕. Compared with the healthy control group, the serum levels of Cys-C and Hcy in the above four groups of different syndromes had a tendency of gradual elevation in the sequence as follows: deficiency of both Qi and Yin, Qi deficiency and blood stagnation, heart and kidney Yang deficiency and flooding due to Yang deficiencygroups〔Cys-C(mg/L):1.02±0.27,1.09±0.31,1.32±0.22, 1.59±0.25; Hcy(μmol/L): 14.94±2.20, 17.66±3.04, 19.79±3.48, 22.96±5.31〕, and the elevation in levels of flooding due to Yang deficiency group was the most prominent compared with that in other groups(P<0.05). The correlation analyses showed that different types of TCM syndrome in patients with CHF were positively correlated with the levels of Cys-C and Hcy(r1=0.73,r2=0.79,bothP<0.05).ConclusionThe changes of serum Cys-C and Hcy levels are consistent with the evolution of regular pattern of TCM syndrome differentiation in patients with CHF, and these two markers can be regarded as the objective indicators of TCM syndrome differentiation of CHF.

目的 观察辨证使用参附、参麦注射液对扩张型心肌病心力衰竭患者心功能的影响.方法 采用病例对照研究方法,将93例扩张型心肌病心衰患者分为3组,符合心阳亏虚型进入A组,在西医常规治疗基础上加用参附注射液.符合气阴两虚型进入B组,在西医常规治疗基础上加用参麦注射液.C组仅用西医常规抗心衰治疗.观察3组患者治疗前后血压、心率、6分钟步行试验、血B型脑钠肽(BNP)浓度、高敏C反应蛋白的变化.结果 治疗2周后,A组、B组在控制血压与心率,降低B型脑钠肽与高敏C反应蛋白,改善6分钟步行试验方面明显优于C组,差异有统计学意义(P＜0.05).结论 对扩张型心肌病心衰患者辨证联合使用参附、参麦注射液,可降低脑钠肽与高敏C反应蛋白,改善亚极量运动能力,有效改善心功能.可能机制为控制心率、调节心脏神经内分泌、增强心肌收缩力等.

Thymosin beta4 (Tβ4) has multi-functional roles in angiogenesis and arteriogenesis, but little is known about its mechanism. The Notch signaling pathway is important in regulation of angiogenic behavior of endothelial cells, in addition to vascular endothelial growth factor (VEGF). Whether, Tβ4 regulates angiogenesis through Notch signaling pathway is not clear. In this article, we evaluated the effect of Notch signaling in Tβ4-induced angiogenesis in human umbilical vein endothelial cell (HUVEC). Our results revealed that Tβ4 increased Notch1 and Notch4 expression in a dose and time-dependent manner. The inhibition of Notch1 or Notch4 with siRNA or the Notch receptor inhibitor DAPT significantly prevented Tβ4-induced HUVEC tube formation and lymphocyte transendothelial migration. The inhibition of Notch1 or Notch4 also blocked Tβ4-induced VEGF and HIF-1α expression. VE-cadherin is the major endothelial adhesion molecule in the control of angiogenesis. Tβ4 significantly reduced VE-cadherin expression levels in HUVEC, while the inhibition of Notch signaling prevented Tβ4-induced VE-cadherin down-regulation. The results of this study suggest that Tβ4 induces HUVEC angiogenesis through Notch signaling pathway.

目的:观察参附注射液联合曲美他嗪对扩张型心肌痛心力衰竭患者的临床疗效.方法:采用前瞻性研究方法,将79例扩张型心肌痛心力衰竭患者随机分为治疗组与对照组,两组均采用西医常规治疗,对照组和西医常规治疗基础上加曲美他嗪,治疗组在对照组治疗基础上加用参附注射液,2周为1个疗程.观察两组患者治疗前后明尼苏达心衰生活质量评分、血压、心率、6分钟步行试验、血脑钠肽(BNP)浓度及射血分数等的变化.结果:治疗组在明尼苏达心衰生活质量评分、控制血压与心率、改善心功能、降低脑钠肽、提高射血分数等方面均明显优于对照组.结论:应用参附注射液联合曲美他嗪治疗扩张型心肌病心力衰竭患者,能明显改善症状,有效改善心功能.

Objective: B-type natriuretic peptide (BNP) was recently demonstrated to be a potential stimulator of angiogenesis and arteriogenesis. The correlation between BNP level and collateral formation in patients with coronary artery disease (CAD) has not been reported.Methods and results: The study included 311 consecutive patients who underwent coronary angiography were divided into three groups according to coronary angiography and collateral formation: normal group (100 patients with normal coronary angiographic findings); poor collateral group (116 patients with at least one coronary stenosis of >= 75% without visible collateral circulation); and good collateral group (95 patients with at least one coronary stenosis of >= 75% with well-developed collateral circulation). Collateral score was analyzed using the Cohen-Rentrop classification. Plasma BNP levels were 45.77 +/- 4.66 pg/ml, 116.40 +/- 28.15 pg/ml, and 254.20 +/- 42.85 pg/ml for patients in normal, poor collateral, and good collateral groups, respectively. Plasma BNP levels in the latter were significantly higher than in the normal group (p < 0.01) and poor collateral group (p < 0.05). There were no significant differences between the good collateral group and poor collateral group when compared with left ventricular ejection fraction (LVEF), left ventricular dimensions at end diastole (LVEDd), age, severity of angiographic disease, and other cardiovascular risk factors. After adjustment in the multiple ordinal logistic regression model, plasma BNP levels showed a strong independent association with collateral Cohen-Rentrop score (chi(2) = 5.636, OR = 1.002, 95% CI 1.000-1.004, p = 0.018).Conclusions: An elevated level of BNP in plasma is independently associated with collateral development; patients with good collaterals tend to have a higher BNP level.

Background Thymosin beta 4 was recently demonstrated to be a potential stimulator of angiogenesis and arteriogenesis in both in-vivo and in-vitro studies. Little is known about the relationship between serum thymosin b4 levels and the extent of coronary collaterals in patients with coronary artery disease.Hypothesis Serum thymosin beta 4 independently predicts the formation of coronary collaterals in patients with coronary artery disease.Materials and methods A total of 190 consecutive patients who underwent coronary angiography were divided into two groups according to coronary angiograms and collateral formation: the poor collateral group consisting of 103 patients with at least more than or equal to one coronary stenosis of greater than or equal to 75% without visible collateral circulation, and the good collateral group consisting of 87 patients with at least more than or equal to one coronary stenosis of greater than or equal to 75% with well-developed collateral circulation. Collateral score was analyzed using the Cohen-Rentrop classification.Results Serum thymosin beta 4 levels were 1024.50 +/- 516.92 ng/ml and 1373.00 +/- 1082.88 ng/ml for patients in the poor collateral group and the good collateral group, respectively. Serum thymosin beta 4 levels in the good collateral group were significantly higher than those in the poor collateral group (P < 0.01). After adjustment in the multiple ordinal logistic regression model, serum thymosin beta 4 levels showed a strong independent association with the collateral Cohen-Rentrop score (chi(2) = 4.45, odds ratio = 1.00, 95% confidence interval: 1.000-1.002, P = 0.035).Conclusion Elevated serum thymosin beta 4 level is independently associated with a significant enhancement in coronary collateralization and patients in the good collateral tend to have a higher thymosin beta 4 level. Coron Artery Dis 22:401-404 (C) 2011 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

ObjectiveIn-vivo studies have shown that hyaluronan (HA) can promote angiogenesis and arteriogenesis, which results in accelerated collateral development. This study is aimed at investigating the association between plasma HA levels and the extent of coronary collaterals, in patients with coronary artery disease. MethodsA total of 253 consecutive patients, who underwent coronary angiography, were divided into three groups according to coronary angiograms: normal group, 81 patients with normal coronary angiographic findings; poor collateral group, 98 patients with at least one coronary stenosis of at least 75%, but without visible collateral circulation; good collateral group, 74 patients with at least one coronary stenosis of at least 75% with well-developed collateral circulation. Plasma HA levels were measured by radioimmunoassay. The correlation between HA levels and the extent of coronary collaterals according to the Cohen–Rentrop classification was calculated by cumulative logits models. ResultsPlasma HA levels were 43.71±2.91, 61.77±4.10, and 131.97±11.76 ng/ml, for patients in the normal, poor collateral, and good collateral groups, respectively. The good collateral group had significantly higher plasma HA levels than the poor collateral (P<0.001) and normal group (P<0.001), whereas there was no significant difference between the normal and poor collateral group. HA levels elevated with increasing Rentrop score, and the cumulative logits model showed a strong graded association between plasma HA levels and the collateral Cohen–Rentrop score (odds ratio=1.021, χ2=17.811, 95% confidence interval: 1.011–1.031, P=0.000). ConclusionThis study suggests that elevated plasma HA levels are associated with a significant enhancement in coronary collateralization. HA may serve as a novel potential biomarker for collateral formation in patients with coronary artery disease.