Hypertensive vascular and cardiac remodeling protection by allicin in spontaneous hypertension rats via CaMK â ¡/NF-κB pathway.

Allicin is the main active component of Traditional Chinese medicine, garlic. It is widely used to treat cardiovascular diseases. Our previous studies have confirmed that allicin significantly reduces blood pressure in Spontaneous Hypertension Rats (SHRs). However, the reports studying the effect of allicin on vascular and cardiac remodeling caused by hypertension are few, with their underlying mechanism not being studied in detail or fully elucidated. In this study, we treated 12-week-old SHRs with allicin for 4 weeks. After 4 weeks, allicin was shown to improve vascular and cardiac remodeling in SHRs, as evidenced by reduced cardiac left ventricular wall thickness, aortic vessel thickness, and reduced proliferating cell nuclear antigen (PCNA) and smooth muscle actin (α-SMA), and increased expression of and smooth muscle 22α (SM 22α). Additionally, allicin reduced serum IL-1ß, IL-6, and TNF-α levels, improved calcium homeostasis in cardiomyocytes, downregulated calcium transportation-related CaMK II and inflammation-related NF-κB and NLRP3, which were observed in smooth muscle cells and cardiomyocytes. Thus, we inferred that allicin protected hypertensive vascular and cardiac remodeling in Spontaneous Hypertensive Rats by inhibiting the activation of the CaMK II/ NF-κB pathway. This study also provided new mechanistic insights into the anti-hypertensive vascular and cardiac remodeling effects of allicin, highlighting its therapeutic potential.

Revealing Calcium Signaling Pathway as Novel Mechanism of Danhong Injection for Treating Acute Myocardial Infarction by Systems Pharmacology and Experiment Validation.

Introduction: Danhong injection (DHI) is a traditional Chinese medicine preparation commonly used in the clinical treatment of acute myocardial infarction (AMI). In this study, the active components of DHI and its mechanism in the treatment of AMI were investigated. Methods: The chemical components of DHI were detected by the ultra-high-performance liquid chromatography-linear trap quadrupole-orbitrap-tandem mass spectrometry (UHPLC-LTQ-Orbitrap-MS/MS), and the targets and pathways of DHI in the treatment of AMI were analyzed by systems pharmacology, which was verified by molecular docking and animal experiments. Results: A total of 12 active components of DHI were obtained, and 158 common targets of component and disease were identified by systems pharmacology. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis results showed that DHI is closely related to the calcium signaling pathway in the treatment of AMI. Molecular docking showed that the key target protein has good binding affinity to related compounds. The experimental results showed that compared with the model group, LVAWs, EF, and FS significantly (p < 0.05) increased in the DHI group. The percentage of myocardial infarction significantly (p < 0.01) decreased, both in the ventricular and total cardiac regions, and the pathological damage of myocardial tissue also decreased. In addition, the expression of the protein CaMK II decreased (p < 0.01) and the expression of SERCA significantly increased (p < 0.01). Conclusion: This study revealed that ferulic acid, caffeic acid and rosmarinic acid could inhibit AMI by regulating PLB, CaMK II, SERCA, etc. And mechanistically, calcium signaling pathway was critically involved. Combination of systems pharmacology prediction with experimental validation may provide a scientific basis for in-depth clinical investigation of the material basis of DHI.

Bacterial endophytes from Lycoris radiata promote the accumulation of Amaryllidaceae alkaloids.

Lycoris radiata is the major source of Amaryllidaceae alkaloids, having various medicinal activities. However, the low content of these alkaloids in planta limits their pharmaceutical development and utilization. In this study, the ability of bacterial endophytes to enhance the accumulation of five important Amaryllidaceae alkaloids was investigated. A total of 188 bacterial endophytes were isolated from L. radiata and their composition and diversity were analyzed. Fourteen ones were demonstrated to significantly increase the concentration of the alkaloids of interest in different organs, up to 11.1-fold over the control level, with no adverse influence on the plant growth. An additional 3 bacterial endophytes were found to significantly increase the dry weight of L. radiata with no adverse influence on the concentration of the alkaloids in planta, so the total yield of alkaloids in planta was increased up to 2.4-fold over the control level. Considering the plant growth-promoting abilities of these bacterial endophytes, it is speculated that the indole-3-acetic acid and siderophore secreted by them, combined with their nitrogen fixation ability, may contribute to the enhanced plant growth and the increased alkaloid accumulation in L. radiata. To our knowledge, this work is firstly defining the diversity of culturable bacterial endophytes in L. radiata and determining which species promoted the accumulation of Amaryllidaceae alkaloids. It provides several valuable bacterial inoculants that can be further applied to improve alkaloid production in L. radiata and broadens our understanding of the interactions between a medicinal plant and the bacterial endophytes.

Toxoflavin Produced by Burkholderia gladioli from Lycoris aurea Is a New Broad-Spectrum Fungicide.

Fungal infections not only cause extensive agricultural damage but also result in serious diseases in the immunodeficient populations of human beings. Moreover, the increasing emergence of drug resistance has led to a decrease in the efficacy of current antifungals. Thus, screening of new antifungal agents is imperative in the fight against antifungal drug resistance. In this study, we show that an endophytic bacterium, Burkholderia gladioli HDXY-02, isolated from the medicinal plant Lycoris aurea, showed broad-spectrum antifungal activity against plant and human fungal pathogens. An antifungal ability assay indicated that the bioactive component was produced from strain HDXY-02 having an extracellular secreted component with a molecular weight lower than 1,000 Da. In addition, we found that this new antifungal could be produced effectively by liquid fermentation of HDXY-02. Furthermore, the purified component contributing to the antifungal activity was identified to be toxoflavin, a yellow compound possessing a pyrimido[5,4-e][1,2,4]triazine ring. In vitro bioactivity studies demonstrated that purified toxoflavin from B. gladioli HDXY-02 cultures had a significant antifungal activity against the human fungal pathogen Aspergillus fumigatus, resulting in abolished germination of conidia. More importantly, the growth inhibition by toxoflavin was observed in both wild-type and drug-resistant mutants (cyp51A and non-cyp51A) of A. fumigatus Finally, an optimized protocol for the large-scale production of toxoflavin (1,533 mg/liter) has been developed. Taken together, our findings provide a promising biosynthetic resource for producing a new antifungal reagent, toxoflavin, from isolates of the endophytic bacterium B. gladioliIMPORTANCE Human fungal infections are a growing problem associated with increased morbidity and mortality. Moreover, a growing number of antifungal-resistant fungal isolates have been reported over the past decade. Thus, the need for novel antifungal agents is imperative. In this study, we show that an endophytic bacterium, Burkholderia gladioli, isolated from the medicinal plant Lycoris aurea, is able to abundantly secrete a compound, toxoflavin, which has a strong fungicidal activity not only against plant fungal pathogens but also against human fungal pathogens Aspergillus fumigatus and Candida albicans, Cryptococcus neoformans, and the model filamentous fungus Aspergillus nidulans More importantly, toxoflavin also displays an efficacious inhibitory effect against azole antifungal-resistant mutants of A. fumigatus Consequently, our findings provide a promising approach to abundantly produce toxoflavin, which has novel broad-spectrum antifungal activity, especially against those currently problematic drug-resistant isolates.

Molecular Cloning and Characterization of a meta/para-O-Methyltransferase from Lycoris aurea.

O-methyltransferases (OMTs) have been demonstrated to play key roles in the biosynthesis of plant secondary metabolites, such as alkaloids, isoprenoids, and phenolic compounds. Here, we isolated and characterized an OMT gene from Lycoris aurea (namely LaOMT1), based on our previous transcriptome sequencing data. Sequence alignment and phylogenetic analysis showed that LaOMT1 belongs to the class I OMT, and shares high identity to other known plant OMTs. Also, LaOMT1 is highly identical in its amino acid sequence to NpN4OMT, a norbelladine 4'-OMT from Narcissus sp. aff. pseudonarcissus involved in the biosynthesis of Amaryllidaceae alkaloids. Biochemical analysis indicated that the recombinant LaOMT1 displayed both para and metaO-methylation activities with caffeic acid and 3,4-dihydroxybenzaldehyde, and showed a strong preference for the meta position. Besides, LaOMT1 also catalyzes the O-methylation of norbelladine to form 4'-O-methylnorbelladine, which has been demonstrated to be a universal precursor of all the primary Amaryllidaceae alkaloid skeletons. The results from quantitative real-time PCR assay indicated that LaOMT1 was ubiquitously expressed in different tissues of L. aurea, and its highest expression level was observed in the ovary. Meanwhile, the largest concentration of lycorine and galanthamine were found in the ovary, whereas the highest level of narciclasine was observed in the bulb. In addition, sodium chloride (NaCl), cold, polyethylene glycol (PEG), sodium nitroprusside (SNP), and methyl jasmonate (MeJA) treatments could significantly increase LaOMT1 transcripts, while abscisic acid (ABA) treatment dramatically decreased the expression level of LaOMT1. Subcellular localization showed that LaOMT1 is mainly localized in cytoplasm and endosome. Our results in this study indicate that LaOMT1 may play a multifunctional role, and lay the foundation for Amaryllidaceae alkaloid biosynthesis in L. aurea.

Effect of Allicin against Ischemia/Hypoxia-Induced H9c2 Myoblast Apoptosis via eNOS/NO Pathway-Mediated Antioxidant Activity.

Allicin (2-propene-1-sulfinothioic acid S-2-propenyl ester, diallyl thiosulfinate) is the main biologically active ingredient in garlic. The present study investigated the protective effect of allicin against cardiomyocyte apoptosis that was induced by ischemia in vitro and the potential molecular mechanisms that were involved in this antiapoptotic effect. The results indicated that allicin increased H9c2 cell activity and attenuated the rate of apoptosis that was induced by ischemia/hypoxia. Intracellular calcium concentrations significantly decreased in the allicin-treated groups. Bax expression significantly decreased, and Bcl-2 expression increased in allicin-treated rats. Nitric oxide blockade significantly inhibited these effects. Allicin also increased the activity of SOD and NO release and decreased MDA levels. Allicin significantly increased the expression of eNOS, Nrf2, and HO-1 proteins. Collectively, these findings demonstrate that allicin protects H9c2 cells against apoptosis, and this protective effect appears to occur via eNOS/NO pathway-mediated antioxidant activity.

[Developing traditional Chinese medicine injection is the need for curing sickness to save patients].

Safety issues of traditional Chinese medicine injections has been heated debate. There are two diametrically opposed views: it should be used reasonable and developed healthily or be forbidden to use. Some people have many misunderstandings and prejudices about the safety of traditional Chinese medicine injections. Compared with western medicine,traditional Chinese medicine has its own particularity. Traditional Chinese medicine has complex components. Its research and clinical application is different from western medicine. Adverse reactions of traditional Chinese medicine injections are related to many factors,such as a large number of irrational use,blind use of traditional Chinese medicine injections and western medicine injections,counterfeit and substandard drugs,incorrect methods of intravenous infusion,toxicity of supplementary materials,drug ingredients. Developing traditional Chinese medicine injection is the need for curing sickness to save patients. The purposeful, targeted, organized and planned systematic research of traditional Chinese medicine injections should be strengthened,especially the safety of traditional Chinese medicine. Strengthen supervision and control of rational drug use.Strengthen the examination and approval,supervision and management of all aspects to ensure the safety of patients.

Allicin improves cardiac function by protecting against apoptosis in rat model of myocardial infarction.

OBJECTIVE: To study the effects of allicin on cardiac function and underlying mechanism in rat model of myocardial infarction (MI). METHODS: Ninety-four Wistar rats were randomly assigned to 6 groups (n=14-16 per group): sham control group [underwent thoracotomy without left anterior descending (LAD) occlusion and only received an injection of the same amount of citrate buffer], MI control group (subjected to LAD occlusion and only received an injection of same amount of citrate buffer), positive control group (subjected to LAD occlusion and received an injection of diltiazem hydrochloride at the dose of 1.5 mg/kg), and MI + allicin groups (subjected to LAD occlusion and received an injection of allicin at the doses of 1.2, 1.8, and 3.6 mg/kg). All of the drugs were administered intraperitoneally daily for 21 days. The infarct area was measured by myocardial staining. Hematoxylin-eosin staining was used to observe the pathological changes. Cardiac function parameters were assessed by echocardiography. The myocardial apoptotic index was estimated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining. The expression of Bax and Bcl-2 were detected by quantificational real-time polymerase chain reaction and Western blot. RESULTS: Treatment with allicin could attenuate the myocardial infarct area (P<0.05) and relieve the changes of the myocardium. The left ventricular anterior wall diastolic and systolic thicknesses were increased in the allicin-treated groups (P<0.05), while there was no signifificant difference in the left ventricular posterior wall diastolic and systolic thickness (P>0.05). The left ventricular internal diameter in systole, ejection fraction, fractional shortening, and stroke volume were dramatically elevated in allicin-treated rats (P<0.05). Allicin dose-dependently reduced creatine kinase and lactate dehydrogenase levels (P<0.05). The myocardial apoptotic index was also markedly lowered, and Bax expression was signifificantly decreased, whereas Bcl-2 expression exhibited an opposite trend in allicin-treated rats (P<0.05). CONCLUSION: Allicin appears to exert a cardioprotective effect that may be linked to blocking Bcl-2/Bax signaling pathway-denpendent apoptosis, further improving cardiac function.

[Effect of allicin on myocardial fibrosis after myocardial infarction in rats and its relationship with TGFß/Smads signal transduction].

Allicin is the internationally accepted active substance of garlic, and has cardiovascular protective effect. This research was designed to investigate the effect of allicin on myocardial fibrosis after myocardial infarction and explore the relationship between the effect and TGFß1/Smads signaling pathway. The rat myocardial infarction model were made by ligating the left anterior desending coronary artery. The drugs were administered intraperitoneally 24 h after the operation. After 21 days, the rats were sacrificed and myocardial collagen fibres were observed by Masson staining. The protein expression of Ⅰ, Ⅲ collagen and TGFß1, Smad3, Smad7 in the myocardium was measured by the immunohistochemistry. The results showed that myocardial fibrosis was serious and the expression of Ⅰ, Ⅲ collagen was increased in model group. After treatment with allicin, the myocardial fibrosis could be relieved markedly, and the expression of collagen was down-regulated. Meanwhile, TGFß1 and Smad3 in heart tissue could be down-regulated and Smad7 could be up-regulated in allicin groups. So allicin may exhibit anti-myocardial fibrosis effect on rats, and the mechanism of this is related to TGFß/Smads signal transduction.

[Effect of paeonol on blood pressure and blood flow in artery of spontaneously hypertensive rats and its mechanisms related on vasomotion].

Previous studies have shown that paeonol can antagonize acute myocardial ischemia and infarction in rat. This study further researched the effects of paeonol on blood pressure and blood flow in the artery of spontaneously hypertensive rats and its mechanisms related on vasomotion. Firstly, thirty spontaneously hypertensive rats were randomly divided into spontaneously hypertensive control group and paeonol-treating groups of high dose and low dose, and also, the other ten Wistar rats as healthy control group. Before and after the intraduodenal administration of the drug, arterial blood pressure was measured by carotid artery and blood flow through the renal artery and carotid artery in vivo were measured by animal flowmeter. The same volume of solvent was given to the spontaneously hypertensive control group and the healthy control group, and the other operations were same. In order to further study the effect of paeonol on vasomotor function, the superior mesenteric artery, renal artery and coronary artery of the spontaneously hypertensive rat were removed and separated, precontracted by a certain concentration of potassium chloride (KCl) and 5-serotonin (5-HT) respectively, and dilatory responses were assessed by cumulative addition of paeonol. Results showed that after duodenal one-time delivery of paeonol, the blood pressure significantly lowered, the renal arterial blood flow and the carotid arterial blood flow significantly increased in spontaneously hypertensive rat. And also, paeonol relaxed the mesenteric artery, renal artery and the coronary artery of spontaneously hypertensive rat in a concentration-dependent manner. These results indicated that the effect of paeonol on decreasing arterial blood pressure and increasing the arterial blood flow was related to its vasodilative effect.

[Academician Li Lianda talking about doctors doing scientific research].

At present, Chinese medical field faces with an important problem of how to correctly handle the relationship between medical and scientific research. Academician Li Lianda advocates doctors doing scientific research under the premise of putting the medical work first. He points out that there are many problems in the process of doctors doing scientific research at present such as paying more attention to scientific research than medical care, excessively promoting building scientific research hospital, only paying attention to training scientific talents, research direction be flashy without substance, the medical evaluation system should be improved and so on. Medical, scientific research and teaching are inseparable because improving medical standards depends on scientific research and personnel training. But not all doctors need to take into account of medical treatment, scientific research and teaching in the same degree while not all hospitals need to turn into three-in-one hospital, scientific research hospital or teaching hospital. It must be treated differently according to the actual situation.

[Specificity of traditional Chinese medicine's clinical re-evaluation].

Clinical re-evaluation is to verify the drug's safety and effectiveness again,while the drug itself has not been improved. However, due to the complexity of traditional Chinese medicine, ingredients in bulk drugs, prescription, productive processes, quality standards and other aspects need to be enhanced. So improving the quality, safety and effectiveness of traditional Chinese medicine by clinical re-evaluation is also very necessary. Therefore, except for achieving those basic requirements of medicine, it should also be improved on itself and pay full attention to the particularity, then traditional Chinese medicine's clinical re-evaluation will play its due role.

[Comparative on acute myocardial infarction models in beagle dogs and mongrel dogs].

OBJECTIVE: To compare the acute myocardial infarction models in Beagle dogs and mongrel dogs, and study whether the Beagle dog model is sensitive to drug intervention. METHOD: The acute myocardial infarction model of dog was set up through ligation of anterior descending branch of coronary artery in dogs, in order to observe morphological changes of the heart and determine artery length and heart coefficient of exposed anterior descending branch of coronary artery. The epicardium electrocardiogram (sigmaST, N-ST) was used to measure the degree of myocardial ischemia. The quantitative histological assay (nitroblue tetrazolium, N-BT stain) was adopted to determine the area of myocardial infarction. RESULT: There was no significant difference between Beagle dogs and mongrel dogs in terms of sigmaST, N-ST and ischemia area. The diltiazem group of Beagle dogs showed obvious reduction in the ischemia area (P < 0.05 and P < 0.01), with notable decline in sigmaST and N-ST, however, it had no statistical difference compared with the Beagle dog model group. Beagle dogs had clear coronary branches, longer exposed arteries and less difference in organ coefficient, which were suitable for the preparation of the myocardial infarction model, whereas mongrel dogs had irregular coronary branches and exposed arteries, with greater individual difference. CONCLUSION: Beagle dogs are superior to mongrel dogs in the preparation of the acute myocardial infarction model, which is sensitive to for drug intervention.

Paeonol induces vasodilatation in rat mesenteric artery via inhibiting extracellular Ca²âº influx and intracellular Ca²âº release.

OBJECTIVE: To investigate the vasodilative effect of paeonol in rat mesenteric artery and the mechanisms responsible for it. METHODS: Rats were anaesthetized and sacrificed. The superior mesenteric artery was removed, dissected free of adherent tissue and cut into 2.0 mm long cylindrical segments. Isometric tension of artery rings was recorded by a myograph system in vitro. Concentration-relaxation curves of paeonol (17.8 µ mol/L to 3.16 mmol/L) were recorded on artery rings precontracted by potassium chloride (KCl) and concentration-contraction curves of KCl, 5-hydroxytryptamine (5-HT), noradrenaline (NA) or calcium chloride (CaCl2) were recorded in the presence of paeonol (10(-4.5), 10(-3.8), 10(-3.5) mol/L) respectively. And also, concentration-relaxation curves of paeonol were recorded in the presence of different potassium channel inhibitors and propranolol on rings precontracted with KCl respectively. To investigate the role of intracellular Ca(2+) release from Ca(2+) store, the contraction induced by NA (100 µ mol/L) and CaCl2 (2 mmol/L) in Ca(2+) free medium was observed in the presence of paeonol respectively. RESULTS: Paeonol relaxed artery rings precontracted by KCl in a concentration-dependent manner and the vasodilatation effect was not affected by endothelium denudation. Paeonol significant decreased the maximum contractions (Emax) induced by KCl, CaCl2, NA and 5-HT, as well as Emax induced by NA and CaCl2 in Ca(2+) -free medium, suggesting that paeonol dilated the artery via inhibiting the extracellular Ca(2+) influx mediated by voltage-dependent calcium channel, and receptor-mediated Ca(2+)-influx and release. Moreover, none of glibenclamide, tetraethylammonium, barium chlorded and propranolol affected the paeonol-induced vasodilatation, indicating that the vasodilatation was not contributed to ATP sensitive potassium channel, calcium-activated potassium channel, inwardly rectifying potassium channel, and ß-adrenoceptor. CONCLUSION: Paeonol induces non-endothelium dependent-vasodilatation in rat mesenteric artery via inhibiting voltage-dependent calcium channel-mediated extracellular Ca(2+) influx and receptor-mediated Ca(2+) influx and release.

[Experimental study on anaphylactoid reactions induced by different components of shengmai injection (new production process) on Beagle dogs].

OBJECTIVE: To evaluate the sensitization effect of different components of Shengmai injection (new production process) on Beagle dogs. METHOD: Beagle dogs were randomly divided into 7 groups, 3 in each group. Each group was respectively injected with 5% glucose injection, Ginseng Radix et Rhizoma Rubra extract, Ophiopogonis Radix extract, Schisandrae Chinensis Fructus extract, Schisandrae Chinersis Fructus distillate, Shengmaifang, 0.2% tween 80. The changes of each dog were observed from injection before until 24 hours after injection, and the response level was determined according to the severity of the symptoms. Blood samples were collected before injection and at 10 min after injection for measuring histamine content in plasma by ELISA. Sensitization of the injection was comprehensively determined by combined the response level of symptoms and the histamine level. RESULT: One dog of Ginseng Radix et Rhizoma Rubra extract group showed untypical symptoms of anaphylactoid reactions, and serum histamine of two dogs increased more than doubled. The Beagle dogs administrated with 0.2% tween 80 showed typical symptoms of anaphylactoid reactions, while there was no significant increase of serum histamine. Other groups were observed with no typical anaphylactoid reactions. CONCLUSION: The sensitization effect of Shengmai injection (new production process) may be associated with Ginseng Radix et Rhizoma Rubra extract and 0.2% tween 80.

[Experimental study on anaphylactoid reactions induced by shengmai injection (new production process) on Cynomolgus monkey].

OBJECTIVE: To compare the anaphylactoid effect of old Shengmai injection and new Shengmai injection on Cynomolgus monkey. METHOD: Cynomolgus monkeys were randomly divided into 4 groups, and were respectively injected with 5% glucose injection, old Shengmai injection, new Shengmai injection, positive control drug. The changes of each monkey were observed from injection before until 24 hours after injection, and the response level was determined according to the severity of the symptoms. Blood samples were collected before injection and at 10 min after injection for measuring histamine content in plasma. Blood pressure and heart rates were detected before injection and at 10 min after injection. Sensitization of the injection was comprehensively determined by combined the response level of symptoms and the histamine level. RESULT: The Cynomolgus monkeys administrated with old Shengmai injection showed typical symptoms of anaphylactoid reactions and the content of serum histamine is not more than doubled. The Cynomolgus monkeys administrated with new Shengmai injection showed untypical symptoms of anaphylactoid reactions and the content of serum histamine did not rise. CONCLUSION: The old Shengmai injection can induce typical anaphylactoid reactions on Cynomolgus monkeys, and the sensitization ability is strong. The symptoms of anaphylactoid reactions induced by the new Shengmai injection appeared later and showed lesser degree with the sensitization lower.

[Experimental study on anaphylactoid reactions of guinea pig induced by danshen injection and its components].

OBJECTIVE: To compare the anaphylactoid effect of Danshen injection and its components on guinea pig. METHOD: Applying active systemic anaphylaxis (ASA) tests, the corresponding experimental injections were administrated to guinea pigs to sensitized, and allergen with double doses was injected to stimulate in the 11 days after the last sensitized. The anaphylaxis situation of guinea pigs was observed. RESULT: Danshen injection and its components are suspicion on guinea pigs, while negative reaction was observed on guinea pigs which injected by the liquid excipients of Danshen injection. CONCLUSION: Danshen injection using the ultrafiltration method still have some antigenic impurities which cannot be removed completely, and this may be one of the reasons for anaphylactic reaction.

[Study on improving the speed and quality of making rat myocardial infarction model by coronary artery ligation].

OBJECTIVE: To establish the methods for improving the speed and quality of making rat myocardial infarction model by coronary artery ligation. METHODS: With the precondition of having no effect on rat myocardial infarction size: (1) To compare the thoracotomy time, the ligation time of the way of coronary artery ligation by tying one knot with the way by tying two knots. (2) To compare the survival rate of making model, the onset time of anesthesia, the awakening time, the effect on myocardial infarction size between using hydration chlorine aldehyde and using sodium pentobarbital. RESULTS: In case of having no significant effect on the myocardial infarction size (P > 0.05), the way of tying one knot could significantly shorten the thoracotomy time and the ligation time (P < 0.01). There was no statistical difference in the onset time of anesthesia or the myocardial infarction size between the model made by sodium pentobarbital and that made by hydration chlorine aldehyde (P > 0.05). But the awakening time of the model made by sodium pentobarbital was obviously shortened (P < 0.01). The clarity rate of myocardial infarction size was improved. CONCLUSIONS: The way of tying one knot could improve the speed of model making. Sodium pentobarbital as the anesthetic for in vivo rat myocardial infarction model could improve the clarity rate of myocardial infarction area.

[Protective effects of paeonol, paeoniflorin and their compatibility on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury].

OBJECTIVE: To observe the protective effects of paeonol, paeoniflorin, and their compatibility on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury. METHODS: Cardiomyocytes from neonatal rats were in vitro cultured and injured by a hypoxia of 2.5 - 5 h and a following 2-h reoxygenation. To observe the effects of paeonol and paeoniflorin, four doses of 100, 75, 50 and 25 mg/L were respectively set up. And to observe the compatibility of paeonol and paeoniflorin, five doses were set up as follows: paeonol 40 and 20 mg/L, paeoniflorin 40 and 20 mg/L, compatibility of paeonol 20 mg/L and paeoniflorin 20 mg/L. The above drugs were incubated with cardiomyocytes during the hypoxia and reoxygenation period respectively. No drug intervention was given to the model group, while no modeling was given to the normal control group. The transudatory creatine kinase (CK), lactate dehydrogenase (LDH), and malondialdehyde (MDA) in the culture medium were determined after the hypoxia period and the reoxygenation period respectively, and the total outleakage and the leakage inhibition ratio during the whole procedure were calculated. Results of each group were got from parallel operations for 5 times. RESULTS: Compared with the normal control group, the MDA leakage increased 2.5 h after hypoxia, the leakage and the total outleakage of CK, LDH, and MDA all significantly increased 3 and 5 h after hypoxia, and 2 h after reoxygenation. The leakage inhibition ratio of each index decreased with statistical difference (P<0.01, P<0.05). Compared with the model group, the leakage and the total outleakage of LDH and MDA both decreased in the high dose paeonol group, and the high and middle dose paeoniflorin groups after hypoxia and 2 h after reoxygenation (P<0.01, P<0.05), and the leakage inhibition ratio of each index increased (P<0.01, P<0.05). However, the leakage and the total outleakage of CK decreased in the low dose and the extreme low dose paeonol groups only 2 h after reoxygenation (P<0.01, P<0.05), while the leakage inhibition ratio of CK increased (P<0.01). The leakage and the total outleakage of LDH decreased in the extreme low dose paeoniflorin group only 2 h after reoxygenation (P<0.01), while the leakage inhibition ratio of LDH increased (P<0.01). The effects of their compatibility showed no significant difference (P>0.05). CONCLUSIONS: Paeonol, paeoniflorin, and their compatibility all have remarkable protective effects on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury. There was no significant synergistic effect when paeonol was used with paeoniflorin together.

[Experimental model of histamine-induced anaphylactoid reaction on beagle dogs].

OBJECTIVE: To establish animal model of histamine-induced anaphylactoid reactions on Beagle dogs through intravenous injection of histamine phosphate injection. METHOD: Anaphylactoid reactions was determined according to the changes of praxiology and blood pressure of Beagle dogs after one intravenous injection of histamine phosphate injection. RESULT: It showed that typical anaphylactoid reactions be caused by histamine phosphate injection on Beagle dogs, and the response of the high-dose group was more obvious than that of the low-dose group. CONCLUSION: Intravenous injection of histamine phosphate injection could simulate Beagle dogs to bring typical clinical anaphylactoid reaction, that could be used as a sensitive animal model to evaluate anaphylactoid reaction of traditional Chinese medicine injections (TCMIs) in the pre-clinical experiments of both TCMIs and their composition.