Development of virulent heat-evil-induced thrombosis animal model / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a virulent heat-evil-induced thrombosis animal model, and provide a rational animal model for pathogeny and pathogenesis research of thrombosis-related diseases, anti-thrombosis activity screening and pre-clinical studies of CAHT formula.</p><p><b>METHOD</b>SD rats were pretreated with carrageenin (Ca) intraperitoneal injection, followed by intravenous injection of endotoxin (LPS from E. coli O111:B4) 50 microg x kg(-1) 16 h later. Thrombosis in rat tails were observed during 12-24 h after injection of LPS. The inflammatory mechanism of this model were investigated by analyzing serum level of TNF-alpha, IL-6, TXB2 and 6-keto-PGF 1alpha, CD11b/CD18 expression of white blood cells (WBC) and P-selectin expression of vessel walls.</p><p><b>RESULT</b>In LPS/Ca model group, thrombosis can be clearly observed in the distal part of rat tails after 12-24 h of LPS/Ca treatment. High level of TNF-alpha and IL-6 can be measured in serum. The expression of CD11b/CD18 in WBC and P-selectin in vessel endothelium significantly increased and the number of WBC in peripheral blood markedly decreased shortly after LPS/Ca treatment. The adherence of white blood cells to vessel endothelium which can be seen by microscope mainly contributed to the decrease of WBC. The results indicated that there was obvious inflammation after treatment with LPS/Ca, suggesting that inflammation was the key mechanism for this model.</p><p><b>CONCLUSION</b>This model was developed through treatment of LPS in combination with Ca, of which LPS is considered to be an exotic virulent heat-evil in TCM, while the inflammatory molecules produced in this model, such as TNF-alpha, IL-6, CD11b/CD18 and P-selectin belong to internal virulent heat-evils, so this animal model consists of pathogeny and pathogenesis of virulent heat-evils. virulent heat-evil.</p>

Pharmacodynamic and toxicologic comparative study of crude and processed radix aristolochice / 中国中药杂志

<p><b>OBJECTIVE</b>To study the accumulated toxic action to bandicoot of aqueous extract of crude and processed Radix Aristolochice and the pharmacodynamic action of aqueous and alcoholic extract of crude and processed Radix Aristolochice.</p><p><b>METHOD</b>The LD50 of acute toxicity to mice and chronic accumulated toxicity to bandicoots of crude and processed Radix Aristolochice were observated. Intestinal and myokinetic influence of normal and revulsive hyperactive gastrointestinal motility of mice induced by neostigmine were observated by giving aqueous extract and alcoholic extract of crude and processed Radix Aristolochice. Relieving pain and eliminating inflammation to mice also were observated.</p><p><b>RESULT</b>The LD50 of aqueous extract of crude and processed Radix Aristolochice were 146. 45, 846.06 g X kg(-1) (equivalently to crude drug) respectively by intragastric administration. Bandicoot' general condition, peripheral blood, serum, organic coefficient, histopathologic examination weren't obvious changes after 1 month administrating aqueous extract of crude and processed drug in three dose. Serum indicators-urea nitrogen, cholesterol total, alkaline phosphatase manifestly were heightened and some animals'hepatic cells, nephric tubules and mucosa emerged differently damage at histomorphology by giving crude high dose after 2 months. Above organs emerged different damage in crude middle and high dose and processed high dose after 3 months and serum indicators- creatinine, urea nitrogen manifestly were increased, the coefficients of liver, kidney and gaster manifestly were heightened. However, the toxicity of identical dose processed product was lower than that of crude one. Aqueous extract and alcoholic extract of crude and processed Radix Aristolochice could obviously inhibite normal and revulsive hyperactive gastrointestinal motility by neostigmine of mice, relieve pain in mouse, stretching and heat stimulation models and inhibite dimethyl benzene-induc mouse, auricle inflammation. Pharmacodynamic action wasnt obvious difference in same dose of crude product and processed one.</p><p><b>CONCLUSION</b>Acute toxicity and chronic accumulated toxicity are stepped down after giving processed Radix Aristolochice, but pharmacodynamic effect wasn t lower. In pharmacodynamic effect, aqueous extract can't compare with alcoholic extract in same dose.</p>

Comparative research of thrombosis animal model induced byendotoxin in combination with carrageenan in different animals / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the development of thrombosis animal model induced by endotoxin(LPS) in combination with carrageenan (Ca) in different animals.</p><p><b>METHOD</b>Two species of rats (SD and Wistar) and three species of mice (Kunming, ICR and Balb/c mice) were employed in the study. The animals of each species were randomly divided into control group and model group (LPS/Ca treatment). The animals in the model group were pretreated with Ca ip at the doses of 25 mg x kg(-1) for rats and 150 mg x kg(-1) for mice, and then treated by LPS iv sixteen hours later, while in the control group were given normal saline (NS). Thrombosis in tails was observed at 24 h after LPS iv. Hematologic parameters were tested for all the animals from each species, and the blood concentration of TNFalpha and IL-6 at different time in SD and Wistar rats were measured.</p><p><b>RESULT</b>LPS/Ca combinatory treatment could induce thrombosis animal model in all five animal species, and the thrombus could be clearly observed on the tails. All species had the similar change in hematologic parameters characterized as the significant decrease of white blood cells and platlets. Inflammatory factors TNFalpha and IL-6 could be largely induced in blood of both SD and Wistar rats at 2 h after LPS iv, but both inflammatory factors only transitorily exist in blood at the early stage of thrombosis model formation.</p><p><b>CONCLUSION</b>LPS/Ca combinatory treatment can successfully induce thrombosis animal model in all tested animal species, and thus this model has extensive animal candidates. The secretion of a large amount of inflammatory factors plays a crucial role in the formation of thrombosis animal model.</p>

Development of an animal model of blood stasis syndrome and thrombosis / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an animal model of thrombosis and blood stasis syndrome in rats by using lipopolysaccharide (LPS) in combination with carrageenan (Ca).</p><p><b>METHOD</b>SD rats in control group were randomly divided into control group and model group (LPS/Ca treatment). The rats in model group were firstly treated with Ca ip, and followed by LPS iv sixteen hours later. The rats in control group were given normal saline (NS). The moment of LPS iv was served as 0 h for the observation. The ear microcirculation, blood rheology parameters (whole blood viscosity etab, plasma viscosity etap and platelet aggregation PA), cruor parameters (thrombin time TT, prothrombin time PT, and partial thromboplastin time APIT) and inflammation factors (TNFalpha, IL-6) were observed at different time after treatment.</p><p><b>RESULT</b>LPS/Ca combinatory treatment can induce a stable and repeatable thrombosis animal model. The thrombus can be observed on the tails of rats by naked eyes, and can be quantitatively measured without necessary of autopsy. Obstacle in microcirculation, increase in whole blood viscosity (etab) and a change of platelets aggregation (PA) rate were observed after LPS/Ca treatment. Cruor parameters were significantly prolonged due to large consumption of cruor factors and platelets. The concentration of inflammation factors TNFalpha and IL-6 in blood was obviously increased at the early stage of the model. The results indicate that this animal model has the characteristics of blood stasis syndrome caused by pyrogen and toxin accompanied by thrombosis.</p><p><b>CONCLUSION</b>LPS/Ca combinatory treatment can induce a easily practicable and repeatable animal model characterized as thrombosis and blood stasis syndrome</p>

Inhibitory effect of egg white lysozyme on ceftazidime-induced release of endotoxin from Pseudomonas aeruginosa / 药学学报

<p><b>AIM</b>To investigate the inhibitory effect of egg white lysozyme (LZM) on ceftazidime (CFT)-induced release of endotoxin from Pseudomonas aeruginosa.</p><p><b>METHODS</b>P. aeruginosa PAO1 was inoculated in nutrition broth or diluted rabbit blood free of antibiotics in the presence or absence of LZM and incubated at 37 degrees C on a water bath shaker. beta-Lactam antibiotic, CFT, was added to cultures at 3.5 h (nutrition broth culture) or 5 h (diluted rabbit blood culture) after inoculation. After 3 h of CFT treatment, the supernatants from different bacterial cultures were prepared by centrifuge and the concentrations of endotoxin in the supernatants were measured. The bacterial supernatants were also added to a murine macrophage cell line RAW 264.7 or intravenously injected into carrageenin-sensitized mice. Tumor necrosis factor-alpha (TNF alpha) and nitric oxide (NO) concentrations in RAW 264.7 supernatants or in mouse sera were tested.</p><p><b>RESULTS</b>CFT treatment alone obviously inhibited the growth of P. aeruginosa PAO1 accompanied by strong and rapid bacteriolysis and released relatively high concentration of endotoxin from bacteria both in nutrition broth and in diluted rabbit blood cultures. The bacterial supernatant from CFT treatment alone yielded high concentrations of TNF alpha both in RAW 264.7 cells and in mice and high level of NO in RAW 264.7 cells. Treatment with the combination of LZM and CFT evidently blocked the lysis of bacteria and reduced the release of endotoxin without decreasing bactericidal activity of CFT. TNF alpha and NO productivity of the supernatants prepared from the LZM/CFT combinative treated bacterial cultures were significantly decreased both in RAW 264.7 cells and in mice indicating that the inflammatory activity was reduced.</p><p><b>CONCLUSION</b>LZM can effectively prevent CFT-induced bacteriolysis, endotoxin release and subsequent proinflammatory factor production but without decreasing bactericidal activity of CFT, resulting in the disassociation of bactericidal activity and bacteriolysis. Thus, LZM might be important for preventing endotoxemia in Gram-negative sepsis with the treatment of antibiotics.</p>

Comparative studies on pharmacological effects of angong niuhuang pill with its simplified prescicription / 中国中药杂志

<p><b>OBJECTIVE</b>Based on the therapeutic claims of Angong Niuhuang pill, a series of pharmacodynamic experiments were designed, where pharmacological effects were investigated comparatively with its simplified prescription(realgar and cinnabar are removed from the original pill) as a parallel control in order to explore possible contribution of cinnabar and realgar to pharmacodynamic activities of the pill as a whole.</p><p><b>METHOD</b>Anti-pyretic, sedative, anti-convulsive, and mice-protected effects of the pill and its simplified prescription as a control were observed, respectively, in rabbits with fever induced by typhoid bacillus, in pentobarbital sodium-induced sleeping mice, in mice with convulsion induced by strychnine, or pentylenetetrazole, and in mice with anoxia induced by NaNO2.</p><p><b>RESULT</b>Both the pill and its simplified prescription were found to have Anti-pyretic action and protective effect against the mouse death induced by anoxia, and synergistic interaction with pentobarbital sodium in sedative activity, although neither of them was found to have any effects on the convulsion of mice.</p><p><b>CONCLUSION</b>No significant difference between Angong Niuhuang pill and its simplified prescription was found in the above pharmacodynamic experiments.</p>

Absorption and distribution of mercury and arsenic from realgar and cinnabar of angong niuhuang pill in normal rats and rats with cerebral ischemia / 中国中药杂志

<p><b>OBJECTIVE</b>To study comparatively the characteristics of absorption and distribution of mercury and arsenic from realgar and cinnabar of Angong Niuhuang Pill in normal rats and the rats with cerebral ischemia after oral administration.</p><p><b>METHOD</b>The blood samples and homogenates of liver, kidney and brain were prepared at various intervals after the animals were treated with Angong Niuhuang pill ig. The levels of total mercury and total arsenic in the blood and the organ homogenates were measured with Microwava Accelerated Reaction System and AAs, respectively.</p><p><b>RESULT</b>The blood concentrations of mercury and arseic reached the highest point in normal rats at one hour following single oral dosing of Angong Niuhuang pill. In normal rats, the mercury distribution was characterized by its higher level in blood and kidneys than in other organs, while a higher distribution of arsenic was found in blood than in organs. No difference in the distribution of mercury or arsenic was found between normal rats and rats with cerebral ischemia after the treatment with the pill.</p><p><b>CONCLUSION</b>The highest level of mercury or arsenic in blood occurs at one hour after oral administration of the pill in normal rats. There is a higher distribution of mercury in blood and kidneys, while a higher distribution of the arsenic only in blood. There is no significant difference in the distribution of mercury or arsenic between the normal rats and the ischemic rats.</p>

Reversal of anti-apoptotic action by tetrandrine in human breast carcinoma multidrug-resistant MCF-7 cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study whether the anti-apoptotic action is reversed by tetrandrine in a combination with vincristine in human breast carcinoma MCF-7 multidrug-resistant cells.</p><p><b>METHOD</b>Chromatin condensation was observed by co-staining of fluorescent dyes Hoechst 33342 and propidium iodide; and G1 sub-peak was detected by flow cytometry. Apoptotic cells were detected with TUNEL method. Cellular free ca2+ was determined with Fluo-3 staining method.</p><p><b>RESULT</b>Two types of chromatin condensation were observed after the sensitive and drug-resistant MCF-7 cells were treated with an antitumor drug vincristine 5 mumol.L-1 for 24 h. The number of cell with chromatin condensation was obviously reduced in the drug-resistant cells treated with the same concentration of vincristine, as compared with the sensitive MCF-7 cells. The number of the apoptotic cells was increased by a combination of non-cytotoxic tetrandrine 20 mumol.L-1 and vincristine in both the sensitive and drug-resistant cells, which was confirmed with fluorescent indication and TUNEL method. The increment of introcellular free Ca2+ level in the cells treated with tetrandrine in a combination of vincristine was detected with Fluo-3 staining method.</p><p><b>CONCLUSION</b>The anti-apoptotic action of human breast carcinoma MCF-7 cells can be effectively reversed by tetrandrine.</p>