Effects of Jiaotai Pills on CUMS-induced depression model in mice based on changes of SIRT1 expression in hippocampus / 中国中药杂志

The present study investigated the effects and mechanisms of Jiaotai Pills on depressed mice induced by chronic unpredictable mild stress(CUMS). The CUMS-induced depression model mice were established and the depression behaviors of mice were evaluated by sucrose preference test, open field test, tail suspension test, and forced swimming test. Molecular docking was employed to simulate the interaction of six main active ingredients in Jiaotai Pills with SIRT1. Immunohistochemical staining was used to detect the level of SIRT1 in the hippocampus of mice. Western blot was used to detect the protein expression levels of SIRT1, p-NF-κB p65, NF-κB p65, and FoxO1 in the hippocampus of mice. Enzyme-linked immunosorbent assay(ELISA) kits were used to detect the levels of interleukin(IL)-1β, IL-6, tumor necrosis factor-α(TNF-α), and brain-derived neurotrophic factor(BDNF) in the hippocampus and serum of mice. Biochemical kits were used to detect superoxide dismutase(SOD) activity and malondialdehyde(MDA) and glutathione(GSH) levels in the hippocampus and serum of mice. Liquid chromatography-tandem mass spectrometry(LC-MS/MS) was used to detect the levels of dopamine(DA), 5-hydroxytryptamine(5-HT), and norepinephrine(NE) in the hippocampus and serum of mice. The results showed that the sucrose preference rate, movement distance, and the number of crossing centers were reduced in the model group(P<0.01), and the tail suspension time and swimming immobility time were increased(P<0.01). Molecular docking results indicated good binding of six main active ingredients in Jiaotai Pills to SIRT1. In the hippocampus, the expression level of SIRT1 was reduced(P<0.01), and the levels of p-NF-κB p65/NF-κB p65 and FoxO1 were increased(P<0.01). In the hippocampus and serum, the levels of IL-1β, IL-6, TNF-α, and MDA were increased(P<0.01), and the activity of SOD and the levels of GSH, DA, 5-HT, NE, and BDNF were reduced(P<0.01). The treatment with high-dose Jiaotai Pills increased the sucrose preference rate, movement distance, and the number of crossing centers(P<0.05), reduced tail suspension time and swimming immobility time(P<0.01), elevated hippocampal SIRT1 expression level(P<0.01), decreased hippocampal and serum IL-1β, IL-6, TNF-α, and MDA levels(P<0.01), potentiated SOD activity, and up-regulated GSH, DA, 5-HT, NE, and BDNF levels in the hippocampus and serum(P<0.05, P<0.01) in model mice. In conclusion, the results showed that Jiaotai Pills could improve the depression behaviors of model mice with CUMS-induced depression, and the underlying mechanism was related to the up-regulation of SIRT1 in the hippocampus of mice to exert anti-inflammatory and anti-oxidative stress effects.

Research progress on induction of osteogenic differentiation of mesenchymal stem cells induced by natural small molecule compounds / 中草药

The repair of bone defect has always been a difficulty in the field of orthopedics. The development of stem cell transplantation-based regenerative medicine has brought hope to overcome this disease. Mesenchymal stem cells (MSCs) have strong osteogenic differentiation potentials, which are excellent seed cells for bone tissue engineering. Natural small molecule compounds are characterized by abundant sources, complex structures, and extensive and unique biological activities. In recent years, it has been found to be a natural inducer for the efficient induction of differentiation of mesenchymal stem cells into osteoblasts, and has important application prospects in the treatment of bone lesions by stem cells. In this paper, recent advances in the induction of osteogenic differentiation of mesenchymal stem cells by natural small molecules are reviewed. It is expected to accelerate the clinical application of mesenchymal stem cells in bone lesions and bone tissue engineering.

Effects of placental growth factor on revascularization after acute myocardial infarction in rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of placental growth factor (PlGF) on revascularization after acute myocardial infarction.</p><p><b>METHODS</b>Myocardial infarction model was established by ligation of left anterior descending coronary artery in Wistar rats. Thirty AMI rats were divided into 3 groups with 10 in each group: PlGF, mouse VEGFR-1/Flt-1 antibody, or saline (control group) was injected in the infarcted border zone for each group, respectively. Two weeks later the hemodynamics parameters were measured with a left ventricular needle catheter and a biological signal analysis system; left ventricular remodeling was observed by HE staining; angiogenesis was examined by immunohistochemistry and cardiomyocyte apoptosis rate was detected by TUNEL.</p><p><b>RESULTS</b>The stroke volume, systolic pressure and left ventricular developed pressure in PlGF group were all higher than those in control group (112±7 vs 65.63±8.50 μl, P<0.01; 131.61±9.26 vs 94.84±8.53 mm Hg, P<0.01 and 175.85±11.36 vs 105.50±15.83 mm Hg, P<0.01; respectively). PlGF animals had less ventricular dilation (left ventricular diameter 8.20±0.14 vs 9.25±0.32 mm, P<0.01) and increased left ventricular wall thickness (1.81±0.10 vs 1.35±0.10 mm, P<0.01) compared to controls. The geometry parameter of anti-VEGFR1 and control animals was almost the same. PlGF animals had increased angiogenesis compared to controls (29.44±5.75 vs 15.88±2.42 endothelial cells/high-powered field, P<0.01); the alpha smooth muscle actin (α-SMA) showed that PlGF animal had a higher density of artery than others (25.14±1.83 vs 19.70±2.52 arteries/mm(2), P<0.01), and the density of artery in anti-VEFGR1 group was less than the controls. The apoptosis rate of cardiomyocytes in PlGF animals was significantly lower than that in controls (9.51%±2.75% vs 37.81%±8.74%, P<0.01).</p><p><b>CONCLUSION</b>Regional delivery of PlGF following acute myocardial infarction can improve cardiac function and left ventricular remodeling, enhance angiogenesis and reduce cardiomyocyte apoptosis rate. PlGF may be a potential agent in adjuvant therapy for acute myocardial infarction.</p>

Effect of transplanted mesenchymal stem cells from rats of different ages on the improvement of heart function after acute myocardial infarction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mesenchymal stem cells (MSCs) transplantation is of therapeutic potential after ischemic injury in both experimental and clinical studies. Clinically, elderly patients are more vulnerable to acute myocardial infarction (AMI). But little is known about the characteristics of young donor-derived MSCs transplanted to old patients with AMI. The present study was designed to investigate the effect of transplanted MSCs from rats of different ages on the improvement of heart function after AMI.</p><p><b>METHODS</b>MSCs from Sprague-Dawley (SD) rats were isolated and cultured in vitro. The apoptosis characteristics of MSCs were observed under conditions of ischemia and anoxia. SD rats underwent MI received intramyocardial injection of MSCs from young donor rats (n = 8), old donor rats (n = 8), respectively. AMI control group received equal volume physiological saline. Immunofluorescence was used to observe the differentiation of the grafted cells into cardiomyocytes. Four weeks after cell transplantation, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry for vascular endothelial growth factor (VEGF), VIII-factor immunohistochemistry for vessel density, TUNEL, caspase-3 for cardiomyocyte apoptosis, echocardiography and hemodynamic detection for heart function were performed.</p><p><b>RESULTS</b>The apoptosis rate of the old donor-derived MSCs group was significantly higher than that of the young donor-derived MSCs group under conditions of ischemia and anoxia (P < 0.05). Engrafted MSCs survived, proliferated and differentiated into myocardium-like cells. VEGF gene expression and capillary density in the old donor-derived group were lower than those in the young donor-derived group but higher than those in the control group (P < 0.05). The transplantation of old donor-derived MSCs attenuated apoptosis of cardiomyocytes in the peri-infarct region compared with the control group and the effect was elevated in young donor-derived MSCs (P < 0.05). The heart functions (left ventricle ejection fraction (LVEF), left ventricle fractional shortening (LVFS)) were improved more significantly in the old donor-derived MSCs group than in the control group and the heart function in the young donor-derived MSCs group further improved (P < 0.05).</p><p><b>CONCLUSIONS</b>Young donor-derived MSCs can improve heart function significantly through angiogenesis and decreasing cardiomyocyte apoptosis when transplanted to the infarcted area.</p>

Clinical studies in the effect of operative strategies on the hemodynamics in off-pump coronary artery bypass grafting / 中华外科杂志

<p><b>OBJECTIVE</b>To study the influence of anastomoses sequence on the hemodynamics in off-pump coronary artery bypass grafting (CABG), and to investigate the strategy to stabilize the hemodynamics.</p><p><b>METHODS</b>From March 2005 to March 2007, 67 patients with serious triple-vessel coronary artery lesions (male 45, female 22) with a age range from 44 to 81-years-old were enrolled for off-pump CABG. All the patients underwent left internal mammary artery-left anterior descending branch (LIMA-LAD) anastomose firstly, followed by the foreword anastomose of the other two vessels. According to the anastomose sequence of posterior descending branch (PDA) and obtuse marginal branch (OM), the patients were divided into three groups. Group I (n = 22) did the sequence of PDA-OM-aortic root (Ao). Group II (n = 14) did the sequence of Ao-PDA-OM. Group III (n = 31) did the sequence of Ao-OM-PDA. The hemodynamics markers, including heart rate (HR), central venous pressure (CVP), mean artery pressure (MAP), cardiac index (CI), left ventricular-stroke work index (LVSWI) and right ventricular stroke work index (RVSWI), were analyzed before LIMA-LAD anastomose to serve as baseline. And these markers were re-measured at LIMA-LAD anastomose, immediately after LIMA-LAD anastomose, at PDA anastomose and at OM anastomose.</p><p><b>RESULTS</b>All the patients survived after off-pump CABG. Compared with the baseline, there were no significant hemodynamic changes when doing LIMA-LAD anastomose. When the anastomose finished, the hemodynamic indices improved significantly. When performing OM anastomose, HR and CVP increased significantly, and MAP, CI, LVSWI and RVSWI decreased remarkably in group I and group III, while there were no significant changes of MAP, CI and RVSWI in group II . When performing PDA anastomose, HR and CVP increased significantly, and MAP, CI, LVSWI and RVSWI decreased remarkably in group I and group II, while there were no significant hemodynamic changes in group III except HR.</p><p><b>CONCLUSION</b>Proximal anastomose first when performing off-pump CABG, followed by distal anastomoses of target vessels is beneficial to those who have hard exposure and difficult anastomose of OM and PDA.</p>

Safety and efficacy on cerebral protection with filter devices during carotid artery stenting in Chinese patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To assess the efficacy, safety, procedural success and long-term clinical outcome in patients underwent percutaneous carotid stenting with distal device.</p><p><b>METHODS</b>Percutaneous carotid stents with distal device were implanted to 58 patients with 59 significant (> 75%) carotid artery stenosis (49 men, mean age 68 years) between January 2000 to December 2005. Forty-five out of 58 patients were symptomatic, 35 had coronary artery diseases and 10 had previous strokes.</p><p><b>RESULTS</b>Sixty one carotid stenting were implanted to 59 lesions in 58 patients. Stents with filter devices were successfully implanted in 57 out of 58 (98%) patients. Angioplasty success rate was 100%. Aspirin (300 mg/d) and Clopidogrel (75 mg x 2/d) were administered 3 days prior operation and clopidogrel was discontinued 30 days post stenting and aspirin was continued at dose of 100 mg/d. The percentage of stenotic carotid artery reduced from 85.3% to 6.2% after stenting and the diameter increased from 1.3 +/- 0.9 mm to 5.2 +/- 1.1 mm. Two minor strokes (3.4%) occurred during operation and at 14 days post stenting. All patients were discharged from the hospital after an average of 2.5 days hospitalization. At 14 +/- 2 months follow up, all patients survived and there were 2 asymptomatic restenosis (50% and 70% and the latter underwent successful balloon angioplasty), 2 myocardial infarctions (1 non-Q wave and 1 Q wave myocardial infarction, all underwent successful emergent PCI) and 2 minor strokes occurred at 6 and 8 months post stenting.</p><p><b>CONCLUSION</b>Carotid stenting with distal device appears to be safe and effective in treating patients with carotid artery stenosis.</p>

Safety and efficacy of carotid artery stenting / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the safety and efficacy of carotid artery stenting (CAS).</p><p><b>METHODS</b>We prospectively evaluated the safety and efficacy of 76 carotid artery stenting (CAS) procedures in a consecutive series of 70 Chinese patients. This series represented a high-risk group that included patients with unstable angina, previous ipsilateral CEA, contralateral carotid artery occlusion, post-radiation carotid artery stenosis and other severe co-morbid conditions. The patients had independent neurological examinations before and after the procedure and follow-up cerebral angiography at 6 month.</p><p><b>RESULTS</b>The procedural success rate was 100%. The mean carotid artery stenosis was (82+/-18) % before and (5+/-10) % after the procedure. During the initial hospital period and 30 days after CAS, there was no major stroke but 3 minor strokes (5.7%). No myocardial infarctions or deaths occurred during or within 30 days following CAS. At a mean follow-up of 20+/-12 months, 2 patients (2.8%) had asymptomatic restenosis, and 2 non-Q wave myocardial infarction. Death occurred in 2 cases, but none of them was attributed to a neurological cause. Three patients had minor strokes and no major strokes occurred during the follow-up period.</p><p><b>CONCLUSION</b>Percutaneous carotid artery stenting is feasible for performance in Chinese patients and may lower the restenosis rate.</p>

Upregulating the expression of angiogenesis-related genes by transplanting autologous mononuclear bone marrow cells into myocardial infarction scar and the periphery / 中华心血管病杂志

<p><b>OBJECTIVE</b>To detect the regulation of angiogenic genes involved in the processes of collateral development.</p><p><b>METHODS</b>Myocardial infarction (MI) scar was induced by cryoinjury in New Zealand rabbits. Four weeks after MI, 24 hours before cell transplantation, bone marrow was aspirated from the right thigh bone and mononuclear bone marrow cells (BMCs) were isolated by Ficoll density gradient centrifugation. Then the mononuclear BMCs (n = 8) or IMDM culture medium (n = 8) were transplanted into infarction scar and the periphery. Four weeks after mononuclear BMCs transplantation, DNA microarray analysis was performed to detect the regulation of angiogenesis-related genes in infarction scar and the periphery. And the differences of angiogenic genes expression were compared among several important growth factors by Western blot.</p><p><b>RESULTS</b>DNA microarray analysis showed the detail regulation of genes involved in the angiogenic processes. There were 15 genes upregulated over 3 times in the infarction scar. In addition, we also found more genes are involved in the process of angiogenesis in its periphery than in the infarction scar (40 genes vs. 15 genes). Western bolt analysis further demonstrated that mononuclear BMCs transplantation was capable of increasing the levels of VEGF, FGF and Angiopoietin-I expression in the infarction scar and its periphery, compared with the control group, P < 0.05.</p><p><b>CONCLUSION</b>These findings indicate that the natural angiogenic processes leading to collateral development are extremely complex, since many kinds of bone marrow-derived growth factors involved in the processes after mononuclear BMCs transplantation into infarction sites.</p>

Effects of a plasmid expressing antisense tissue inhibitor of metalloproteinase-1 on liver fibrosis in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>No efficient therapy for liver fibrosis has been available. This study was aimed to provide evidence that the introduction of a plasmid expressing antisense tissue inhibitor of metalloproteinase-1 (TIMP-1) into a rat model of immunologically induced liver fibrosis can result in the increased activity of interstitial collagenase, thus enhancing the degradation of collagen.</p><p><b>METHODS</b>Real-time nested polymerase chain reaction (RT-Nested-PCR) and gene recombination techniques were used to construct a rat antisense TIMP-1 recombinant plasmid that can be expressed in eukaryotic cells. Both the recombinant plasmid and an empty vector (pcDNA3) were encapsulated with glycosyl-poly-L-lysine and injected into rats suffering from pig serum-induced liver fibrosis. The expression of exogenous transfected plasmid was assessed by Northern blot, RT-PCR, and Western blot. Hepatic interstitial collagenase activity was detected using fluorescinisothiocyanate (FITC)-labeled type I collagen. In addition to hepatic hydroxyproline content, hepatic collagen types I and III were detected by immunohistochemical staining, and the stages of liver fibrosis by Van Gieson staining.</p><p><b>RESULTS</b>Exogenous antisense TIMP-1 was successfully expressed in vivo and could block the gene and protein expression of TIMP-1. Active and latent hepatic interstitial collagenase activities were elevated (P < 0.01), hepatic hydroxyproline content and the accumulation of collagen types I and III were lowered, and liver fibrosis was alleviated in the antisense TIMP-1 group (P < 0.01) as compared with the model group.</p><p><b>CONCLUSION</b>The results demonstrate that antisense TIMP-1 recombinant plasmids have some inhibitory effect on liver fibrosis.</p>

Purification and characterization of platelet aggregation inhibitor component from venom of agkistrodon halys pallas / 中国实验血液学杂志

Snake venom proteins,particularly from the viper and elapid families, have been known to contain a number of platelet active components including what cause platelet aggregation or inhibit platelet aggregation. Some of them have potential clinical usefulness for the treatment of human hemorrhagic or thrombotic disease. Agkistrodon halys pallas belonging to viper family is only growing in China. The aim of this study was to purify a human platelet aggregation inhibitor from venom of Agkistrodon halys pallas and determine its biochemical character. Whether a component could inhibit human platelet aggregation was act as a method to follow the tracks of the protein. Crude venom of Agkistrodon halys pallas was loaded onto a DEAE-Sepharose CL-6B chromatography column could gain 6 peaks. A platelet inhibitor with molecular mass of 65 kD on SDS-PAGE, was purified from peak 2 by Sephadex G-75 gel filtration and SP-Sepharose, Mono Q on FPLC. It could inhibit human platelet aggregation induced by ADP, collagen without activities of phospholipase A2, esterase, fibrinogenolytic. It is concluded that a platelet inhibitor can be isolated and purified from venom of Agkistrodon halys pallas and its inhibition of platelet aggregation is does-dependent.

Effects of antisense transforming growth factor beta receptor-II (TGFbetaRII) expressing plasmid on experimental liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effects of antisense transforming growth factor beta receptor-II (TGFbetaRII) expressing plasmid on experimental liver fibrosis.</p><p><b>METHODS</b>RT-Nest-PCR and gene recombinant techniques were used to construct the rat antisense TGFbetaRII recombinant plasmid which can be expressed in eukaryotic cells. Thirty-six male SD rats were randomly distributed into five groups: 10 in experimental liver fibrosis model induced by pig-serum as disease control group; 10 in antisense TGFbetaRII transfection as treatment group; 10 in pCDNA3 transfection as treatment control group and 6 in normal control group. The recombinant plasmid and empty vector (pCDNA3) were encapsulated by glycosyl-poly-L-lysine and then transducted into rats of pig serum-induced liver fibrosis model respectively. Expression of exogenous transfected plasmid was assessed by Northern blot, RT-PCR and Western blot. We also tested ELISA of serum TGF-beta1, the contents of hepatic hydroxyproline, immunohistochemistry of type I and III collagen, and VG staining for pathological study.</p><p><b>RESULTS</b>The antisense TGFbetaRII expressing plasmid could be well expressed in vivo, and could block the mRNA and protein expression of TGFbetaRII in the fibrotic liver induced by pig serum. Its expression also reduced the level of TGF-beta1 [antisense treatment group (23.16+/-3.13) ng/ml, disease control group (32.96+/-3.79) ng/ml; F=36.73, 0.01]. Compared with the disease control group, the contents of hepatic hydroxyproline [antisense treatment group (0.17+/-0.01) mg/g liver, disease control group (0.30+/-0.03) mg/g liver; F=15.48, 0.01] and the deposition of collagens type I and type III decreased in the antisense group (antisense treatment group collagen type I 650.26+/-51.51, collagen type III 661.58+/-55.28; disease control group type I 1209.44+/-116.60, collagen type III 1175.14+/-121.44; F values are 69.87, 70.46, 0.01). And its expression also improved the pathologic classification of liver fibrosis models (0.01).</p><p><b>CONCLUSION</b>The results demonstrate that TGF-beta plays a key role in liver fibrogenesis and the prevention of liver fibrosis by antisense TGFbetaRII recombinant plasmid intervention may be therapeutically useful.</p>