[Development and Quality Evaluation of Evidence-based Clinical Practice Guidelines of Chinese Medicine].

More attentions have been paid to the development of evidence-based clinical practice guidelines (ECPGs) of Chinese medicine (CM). International guideline evaluation instruments such as Appraisal of Guidelines for Research and Evaluation (AGREE I) has been gradually applied in ECPGs quality evaluation of CM. Nowadays, there are some certain methodological defects in partial ECPGs of Chinese medicine, with relatively low applicability and slowly update. It is suggested to establish technical specifications of CM-ECPGs in accordance with the characteristics of CM and international general specification, strengthen the quality evaluation of CM-ECPGs, attach great importance to the regularly update as well as popularization and application of CM-ECPGs.

[Effects of Chinese herbal drug-containing serum on oxidative damage and apoptosis of umbilical vein endothelial cells induced by oxidized low-density lipoprotein].

OBJECTIVE: To investigate the effects of Chinese herbal drug-containing serum, prepared by administration of Chinese herbal medicine for activating blood (Xiongshao Capsule, XS) or for activating blood and detoxifying (Xiongshao Capsule plus Huanglian Capsule, XSHL) in rats, on cell viability, oxidative damage and apoptosis of human umbilical vein endothelial cells (HUVECs) induced by oxidized low-density lipoprotein (ox-LDL). METHODS: Thirty-two rats were randomly divided into 4 groups: control group, positive control group (simvastatin 1.8 mg/kg), activating blood (XS, 0.135 g/kg) group, and activating blood and detoxifying (XS Capsule 0.135 g/kg and Huanglian Capsule 0.135 g/kg, XSHL) group. Corresponding drugs were continuously administered to the rats for 7 days and then drug-containing serum was harvested 1 hour after the last administration. HUVECs isolated from newborn children by collagenase digestion were stimulated by ox-LDL (100 µg/mL) [corrected] and incubated with corresponding drug-containing serum for 24 hours. Untreated HUVECs were also used as a normal control. The morphology and structure of HUVECs were observed by an inverted microscope. Cell viability was measured by methyl thiazolyl tetrazolium method, and cell membrane damage was determined by lactate dehydrogenase (LDH) leakage. Activity of superoxide dismutase (SOD) was examined by spectrophotometry, and content of malondialdehyde (MDA) in the cell lysate was examined by thiobarbituric acid assay. HUVECs were stained with Annexin V-fluorescein isothiocyanate and propidium iodide and analyzed on a flow cytometry to determine apoptosis. RESULTS: Compared with the normal HUVECs, the cell viability and the activity of SOD were significantly decreased while the content of MDA and apoptosis rate were significantly increased after 24-hour ox-LDL stimulation (P<0.01, P<0.05). Simvastatin-, XS-, and XSHL-containing serum significantly promoted the ox-LDL-stimulated HUVEC viability and inhibited early apoptosis (P<0.01, P<0.05), while had no significant effect on LDH leakage. Simvastatin-containing serum and XS-containing serum also showed significant decrease in MDA content and increase in SOD activity, while XSHL-containing serum showed no significant effects. There was no significant difference between the XS-containing serum group and the XSHL-containing serum group. CONCLUSION: Both sera containing XSHL and XS show protective action against the oxidative damage and apoptosis of HUVECs induced by ox-LDL.

[Translational medicine and its application in research & development of integrative medicine].

The background, concept and status quo of translational medicine at home and abroad were introduced systematically in this review, and the application mode of translational medicine in the research and development of Chinese medicine (CM) was analyzed. Targeting the characteristics of CM and the changes in the spectrum of diseases in China, some suggestions were made to strengthen the translational research in CM and integrative medicine.

[Screening, identification and analysis of platelet differential functional proteins in patients with coronary heart disease of blood-stasis pattern].

OBJECTIVE: To seek the key platelet functional proteins (PFPs) for the occurrence of blood-stasis pattern (BP) in patients with coronary heart disease (CHD). METHODS: Peripheral blood platelet protein of 22 patients and 24 healthy person (for control) were extracted respectively in 4 batches for carrying 4 times of the test out. Differential PFPs in samples were screened out by two-dimensional fluorescence difference gel electrophoresis, and identified with matrix-assisted laser desorption/ionization-time of flight mass spectrometry; then the identified proteins were further authenticated by Western-blotting. RESULTS: Thirteen differential PFPs were screened out, and among them the 7 identified by spectrometry were: isoform 1 of integrin alpha- II b, isoform 2 of integrin alpha- II b, actin-cytoplasmic 1, actin-cytoplasmic 2, cDNA FLJ52842, cDNA FLJ55253, and cDNA FLJ43573 fis. Among them isoform 2 of integrin alpha- II b (CD41) and actin-cytoplasmic 2 (Acting) were authenticated successfully. CONCLUSION: CD41 and acting are the possible marker proteins, and the other PFPs might play crucial roles in the occurrence and development of BSS in CHD.

[Current status and problems in developing clinical guidelines for Chinese medicine and integrative medicine].

With the wide application of clinical guidelines and standardization of Chinese medicine (CM), guidelines of CM and/or integrative medicine (IM) were also emerging. By the guideline evaluation instruments such as the Appraisal of Guidelines Research and Evaluation (AGREE) Instrument and Conference on Guideline Standardization (COGS), a preliminary assessment of 11 clinical guidelines for CM and/or IM published before October 2008 was performed. Methodological description of evidence collection or synthesization was absent in most clinical guidelines, and evidence-grading criteria were listed in only one of the eleven guidelines. Inadequate standardization of guideline development, single professional background of guideline developers, and lack of high-grade evidence were the current problems. It was suggested that guideline development group should include individuals from multiple relevant professional fields. Stress should be laid on evidence collection and recommendation grading. Guideline developers should follow the rigorous development methodology of evidence-based guidelines, and the methods for evaluating evidence and grading recommendations should be set up according to the characteristics of medical literature of CM. In addition, more attention should be paid to appraise the quality of clinical practice guidelines of CM and IM.

[Differential gene expression profile in ischemic myocardium of Wistar rats with acute myocardial infarction: the study on gene construction, identification and function].

OBJECTIVE: To construct the differential genes expressed profile in the ischemic myocardium tissue reduced from acute myocardial infarction(AMI), and determine the biological functions of target genes. METHODS: AMI model was generated by ligation of the left anterior descending coronary artery in Wistar rats. Total RNA was extracted from the normal and the ischemic heart tissues under the ligation point 7 days after the operation. Differential gene expression profiles of the two samples were constructed using Long Serial Analysis of Gene Expression(LongSAGE). Real time fluorescence quantitative PCR was used to verify gene expression profile and to identify the expression of 2 functional genes. The activities of enzymes from functional genes were determined by histochemistry. RESULTS: A total of 15,966 tags were screened from the normal and the ischemic LongSAGE maps. The similarities of the sequences were compared using the BLAST algebra in NCBI and 7,665 novel tags were found. In the ischemic tissue 142 genes were significantly changed compared with those in the normal tissue (P<0.05). These differentially expressed genes represented the proteins which might play important roles in the pathways of oxidation and phosphorylation, ATP synthesis and glycolysis. The partial genes identified by LongSAGE were confirmed using real time fluorescence quantitative PCR. Two genes related to energy metabolism, COX5a and ATP5e, were screened and quantified. Expression of two functional genes down-regulated at their mRNA levels and the activities of correlative functional enzymes decreased compared with those in the normal tissue. CONCLUSION: AMI causes a series of changes in gene expression, in which the abnormal expression of genes related to energy metabolism could be one of the molecular mechanisms of AMI. The intervention of the expressions of COX5a and ATP5e may be a new target for AMI therapy.

Effects of propyl gallate on carotid artery thrombosis and coagulation/fibrinolysis system in rats.

OBJECTIVE: To investigate the effects of propyl gallate (PrG) on the thrombus formation time and the coagulation/fibrinolysis system in an experimental carotid artery thrombosis model in rats. METHODS: Fifty SD rats were randomly divided into 5 groups (10 animals/group): the normal group (normal saline 2 mL/kg), the model group (normal saline, 2 mL/kg), the heparin control group (1,250 IU/kg), the low dose PrG group (30 mg/kg), and the high dose PrG group (60 mg/kg). Thirty minutes after intravenous injection of saline or the corresponding drugs, a carotid artery thrombus was induced by continuous electric stimulation in all rats except for those in the normal group. The duration from the initiation of the electric stimulation to the sudden drop in carotid temperature was recorded as the thrombus formation time. Levels of plasma tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) were determined by ELISA. RESULTS: PrG (30 and 60 mg/kg) can prolong the thrombus formation time, but the effect was obviously weaker than that of heparin (P<0.05, P<0.01). Compared with the model group, PrG (30 and 60 mg/kg) elevated the plasma activity of t-PA (both P<0.05) and showed an increasing tendency in elevating the ratio of t-PA/PAI-1 (P>0.05), while it had no significant effect on the level of PAI-1. CONCLUSION: PrG has a certain antithrombotic effect and can slightly regulate the imbalance of the t-PA /PAI-1 ratio.

[Treatment of non-ST-elevation acute coronary syndrome with propyl gallate].

OBJECTIVE: To investigate the therapeutic effects of propyl gallate (PrG) in combination with standard medication on patients with non-ST-elevation acute coronary syndrome (NST-ACS), including unstable angina and acute non-ST-elevation myocardial infarction, and its influences on serum inflammatory marker and platelet activation. METHODS: Fifty-five patients with NST-ACS were randomly assigned to two groups. Accessory to the standard Western medicine, the 27 patients in the tested group treated with PrG and the 28 in the control group with salvia composite (SC), all being medicated for 14 days. Effects on angina pectoris and electrocardiogram were observed. The positive rate and mean fluorescence density (MFI) of GP IIb-IIIa and CD62p expression on platelet surface were detected using flow cytometer; the serum concentration of high sensitive C-reactive protein (Hs-CRP) was determined using ELISA before and after treatment respectively. RESULTS: The therapeutic effects on angina and electrocardiogram between the two groups showed no significant difference. Serum level of Hs-CRP, GP IIb-IIIa MFI and CD62p positive rate were significantly lowered after treatment in both groups (P < 0.05), no significant difference was found between groups, though the lowering of Hs-CRP and GP IIb-IIIa MFI in the tested group displayed a further decreasing trend. CONCLUSION: In combination with standard medication of Western medicine, PrG and SC showed no obvious difference in the therapeutic effect and influences on angina pectoris and electrocardiogram in patients with non-ST-elevation acute coronary syndrome.

[Effects of propyl gallate on serum inflammatory factors and protein expression of COX-2 and ICAM-1 in ischemic myocardium of rats with acute myocardial infarction].

OBJECTIVE: To investigate the effects of Propyl Gallate (PrG) on serum inflammatory factors and protein expression of cyclooxygenase-2 (COX-2) and intercellular adhesion molecule-1 (ICAM-1) in ischemic myocardium of rats with acute myocardial infarction (AMI). METHODS: AMI model was induced by ligating the left anterior descending (LAD) branch of coronary artery in Wistar rats, and the perfect modeling was certified with ST segment elevation by standard limb lead II of electrocardiogram. Rats were randomly divided into 6 groups: Group A of normal rats, Group B of rats through sham operation, Group C of AMI model rats, Group D of model rats treated with high dose PrG (80 mg x kg(-1) x d(-1), via peritoneal injection), Group E of model rats treated with low dose PrG (40 mg x kg(-1) x d(-1), via peritoneal injection), and Group F of model rats treated with aspirin (25 mg x kg(-1) x d(-1), via gastrogavage), all the treatments were given in succession for 7 days. Radioimmunoassay (RIA) was used to determine serum contents of interleukin (IL)-1beta and tumor necrosis factor-alpha (TNF-alpha), immunohistochemistry was used to determine the level of COX-2 and ICAM-1 protein expression in myocardium. RESULTS: Compared with Group B, the serum level of TNF-alpha increased significantly, but not the level of IL-1beta in Group C. Besides, the COX-2 and ICAM-1 protein expressions in ischemic myocardium increased in Group C. All the above-mentioned changes were reversed to certain extent in Group E after treatment. CONCLUSIONS: PrG (40 mg x kg(-1) d(-1)) could decrease the serum level of inflammatory factor TNF-alpha, and slightly inhibit COX-2 and ICAM-1 protein expression in ischemic myocardium of AMI rats.

[Effects of Xuefu Zhuyu Oral Liquid on hemorheology in patients with blood-stasis syndrome due to coronary disease and their relationship with human platelet antigen-3 polymorphism].

OBJECTIVE: To investigate the effects of Xuefu Zhuyu Oral Liquid, a compound traditional Chinese herbal medicine for resolving stagnation, on hemorheology in the patients with blood-stasis syndrome due to coronary disease and their relationship with human platelet antigen-3 (HPA-3) polymorphism of membrane glycoprotein IIb (GPIIb). METHODS: Thirty-two patients with blood-stasis syndrome due to coronary disease were selected in this study. Blood-stasis syndrome scoring was performed and hemorheological parameters were measured in all subjects before and after Xuefu Zhuyu Qral Liquid treatment. The genotypes of GPIIb HPA-3 were determined by TaqMan probe technology. RESULTS: The hemorheology was improved in the patients with blood-stasis syndrome due to coronary heart disease after the treatment. There were significant differences in the whole blood viscosity, deformation index of red blood cells and scores of blood-stasis syndrome between the patients carrying AC plus CC and the patients carrying AA after the treatment (P<0.05). CONCLUSION: Xuefu Zhuyu Oral Liquid can improve clinical symptoms and hemorheology in the patients with blood-stasis syndrome due to coronary heart disease, which is related to GPIIb HPA-3 polymorphism.

Relationship between two blood stasis syndromes and inflammatory factors in patients with acute coronary syndrome.

OBJECTIVE: To investigate the relationship between inflammatory factors and two Chinese medicine (CM) syndrome types of qi stagnation and blood stasis (QSBS) and qi deficiency and blood stasis (QDBS) in patients with acute coronary syndrome (ACS). METHODS: Sixty subjects with ACS, whose pathogenesis changes belongs to qi disturbance blood stasis syndrome, were divided into 2 groups: 30 in the QSBS group and 30 in the QDBS group. The comparative analysis on them was carried out through comparing general information, coronary angiography and inflammatory factors including intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40) and lipoprotein-associated phospholipase A2 (Lp-PLA2). RESULTS: Compared with the QSBS group, Lp-PLA2 and YKL-40 levels in the QDBS group showed no-significant difference (P>0.05); ICAM-1 was significantly higher in the QDBS group than in the QSBS group in the pathological processes of qi disturbance and blood stasis syndrome of ACS (P<0.05). CONCLUSIONS: Inflammatory factor ICAM-1 may be an objective basis for syndrome typing of QSBS and QDBS, which provides a research direction for standardization research of CM syndrome types.

[Acting mechanism of qidan liquid in promoting myocardial angiogenesis in rats after acute myocardial infarction].

OBJECTIVE: To investigate the effect of Qidan Liquid (QL) on myocardial angiogenesis in rats after acute myocardial infarction (AMI) and explore its possible molecular mechanism. METHODS: AMI model was established by ligating the left anterior descending (LAD) coronary artery in Wistar rats, and intervened with QL in high, medium and low doses (QLh, QLm and QL1 group), and isosorbide dinitrate (ID group) respectively. Meanwhile, the model group, the sham group and the normal group were also set up. On the 7th (d7) and 14th day (d14) after modeling, mRNA and protein expressions of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected with RT-PCR and immunohistochemistry, factor VIII expression in endothelial cells was examined, and microvascular density (MVD) in ischemic myocardium was calculated. RESULTS: MVD in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), on d7 and d14, it was lower in the model group than that in the high and medium dose of QL groups and the ID group (P < 0.05). On d7, VEGF protein expression in the model group was higher than that in the normal group (P< 0.05), equivalent to that in the other groups, on d14, it was lower than that in the high and medium dose of QL groups and the ID group (P < 0.05 or P < 0.01). On d7 and d14, the protein expression of bFGF in the model rats increased significantly, higher than that in the normal and the sham group (P < 0.01), it decreased along the time progress between the two time points (P < 0.05), and lower than that in the high dose of QL group and ID group (P < 0.01). The mRNA expressions of VEGF and bFGF were higher in the model group than those in the normal and sham group, but lower than those in all the treated groups on d7 and d14. Comparison of the strips on d7 and d14 among various groups showed that the bFGF mRNA expression in the model group showed a descending trend as the time goes on, but in all the treated groups it was unchanged, while the change of VEGF mRNA expression didn't show any definite tendency. CONCLUSION: QL could up-regulate the protein and gene expressions of VEGF and bFGF persistently and increase MVD in ischemic myocardium to promote the development of collateral circulation in AMI rats.

[Effect of folium panax quinquefolium saponins on apoptosis of cardiac muscle cells and apoptosis-related gene expression in rats with acute myocardial infarction].

OBJECTIVE: To observe the effect of Folium Panax quinquefolium saponins (PQS) on apoptosis of cardiac muscle cells (CMCs) and apoptosis-related gene expression in rats with acute myocardial infarction (AMI). METHODS: Fifty Wistar rats were randomly divided into 5 groups, 40 rats underwent coronary ligation of left anterior descending branch to establish AMI rats model, while to the other 10 rats in the sham-operation group, thoracotomy without coronary ligation was carried out. The AMI model rats in treated groups were treated with high-dosage PQS, low-dosage PQS, captopril respectively for 7 days, and those in the control group were treated with normal saline instead. Then, the apoptotic CMCs were labeled by TUNEL and the expression of Bcl-2 and Fas in cardiac muscle cells were determined by immunohistochemical methods. RESULTS: The apoptotic index of CMCs in the model rats (46.48%) was significantly higher than that in the sham-operation group ( 1.03%, P<0.01 ). CMCs apoptosis rate in the low-dosage PQS group, the large-dosage group and the captopril group was 23.53 %, 17.58 % and 25.17 %, respectively, all were significantly lower than that in the sham-operation group (P<0.01). The expression of Bcl-2 gene was up-regulated and expression of Fas protein was down-regulated in the three treated groups. CONCLUSION: PQS can inhibit CMCs apoptosis in AMI rats, downregulate Fas protein expression and up-regulate Bcl-2 protein expression, and has antagonistic effect in myocardial ischemic injury.

[Effect of ginseng fruit saponins on insulin sensitivity index in high fat-fed rats].

OBJECTIVE: To observe the effect of ginseng fruit saponins (GFS) on insulin sensitivity index in high fat-fed rats. METHODS: An animal model of insulin resistance was established by injecting low dose of streptozotocin (STZ) in high fat-fed rats. Effect of GFS on insulin sensitivity was detected with glucose infusion rate (GIR) by euglycemic hyperinsulinemic clamp technique. RESULTS: The level of fasting blood glucose and insulin in untreated group increased more significantly than that in normal control group (P<0.05, P<0.01), while the index of GIR decreased significantly (P<0.01). As compared with the untreated groupìthe parameters of GFS-treated groups were improved significantly in a dosage-dependent manner (P<0.05, P<0.01). CONCLUSION: GFS can improve experimental insulin resistance in rats.

[Study on interaction of Radix Paeoniae 801 and endothelin-1 by using a piezoelectric quartz crystal biosensor].

OBJECTIVE: To explore the possible target and molecular mechanism of Radix Paeoniae 801 (RP801), an effective ingredient extracted from Radix Paeoniae, the Chinese herbal medicine for activating blood circulation to remove blood stasis, using experimental in vitro method by directly detecting the interaction between RP801 and endothelin-1 (ET-1). METHODS: Piezoelectric quartz crystal biosensor, namely, the quartz crystal microbalance (QCM) was used to detect the specific combining between RP801 and ET-1 by binding avidin to the pre-activated Au surface of electrode of QCM, followed by immobilizing biotinylated ET-1 to it, and adding RP801, then the binding curve was recorded. PBS washing was applied at the end of every steps of combining reaction for dissociate the non-specific absorption. RESULTS: Specific combining of RP801 and ET-1 was found. CONCLUSION: ET-1 could possibly be one of the acting targets of RP801 in the body, that is, RP801 could combine with ET-1 to impede the binding of ET-1 with its receptor, so as to counteract the action of ET-1, dilate blood vessels and inhibit platelet aggregation.