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AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.
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OBJECTIVE@#To investigate the inhibitory effect of agkistrodon halys venom antitumor component-I (AHVAC-I) on vasculogenic mimicry (VM) formation in triple-negative breast cancer MDA-MB-231 cells and explore its possible mechanism.@*METHODS@#CCK8 assay was used to determine the optimal concentration of AHVAC-I for cell treatment based on its halfinhibitory concentration (IC50). MDA-MB-231 cells were treated with different concentrations of AHVAC-I or 5-Fu, and the changes in vasomimetic capacity of the cells were examined using Matrigel assay. The expression levels of matrix metalloproteinase-2 (MMP2) and MMP9 in the treated cells were detected using quantitative PCR and Western blotting.@*RESULTS@#Compared with the control treatment with culture medium, treatment with 5, 10 and 20 μg/mL AHVAC-I significantly reduced vasomimetic ability of MDA-MB-231 cells in a dose-dependent manner (P < 0.01). MMP2 supplementation obviously restored the vasomimetic ability of the cells inhibited by AHVAC-I.@*CONCLUSION@#AHVAC-I inhibits VM formation in triplenegative breast cancer cells in vitro by down-regulating MMP2 production.
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Animals , Humans , Agkistrodon/metabolism , Cell Line, Tumor , Healthy Life Expectancy , Matrix Metalloproteinase 2/metabolism , Neovascularization, Pathologic/metabolism , Triple Negative Breast Neoplasms/metabolism , VenomsABSTRACT
BACKGROUND: Agkistrodon acutus, a traditional Chinese medicine, clinically used in the treatment of rheumatism, tumor, and cardiovascular and cerebrovascular diseases. Due to the unique medicinal value and the difficulty of artificial breeding of Agkistrodon acutus, the supply of Agkistrodon acutus on the market exceeds the demand, and a large number of its adulterants are found on the market. In this study, the cytb gene sequences of Agkistrodon acutus and 9 snakes were compared and analyzed, specific primers were designed, and specific PCR methods were established to detect Agkistrodon acutus medicinal samples on the market. RESULTS: This method was successfully applied to distinguish the snake from other adulterated species, and tested 18 Agkistrodon acutus samples randomly purchased from six cities. Twelve samples were counterfeit and six were genuine. The standard reference material of Agkistrodon acutus was cloned by molecular cloning and sequencing, and the gene sequence difference with other species was significant. It shows that the region could be used as the fingerprint region of the target species. CONCLUSIONS: The proposed method can be used as a species-specific marker and can be highly distinguished from other adulterated snake species, which is helpful to effectively avoid the problem of false sale of Agkistrodon acutus.
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Animals , Polymerase Chain Reaction/methods , Agkistrodon/genetics , Cytochromes b/genetics , Mitochondria/genetics , Snakes , Species Specificity , DNA/analysis , Cloning, Molecular , Medicine, Chinese TraditionalABSTRACT
AIM: To explore whether Agkistrodon Halys venom antitumor component-I (AHVAC-I) affects the migration of gastric cancer cells by human primary gastric cancer-associated fibroblast (GCAFs). METHODS: Tissue block culture and trypsin digestion were used to separate and culture human primary gastric cancer-associated fibroblasts (GCAFs); the GCAFs-CM
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AIM:To explore effect and meaning of Agkistrodon halys venom platelet inhibitor on GPVI expression and hemorheology in rats with acute myocardial ischemia reperfusion injury. METHODS: Thirty matched SD male rats were randomly divided into sham operation group (6 rats), myocardial ischemia-reperfusion injury(MIRI) model group(6 rats) and agkistrodon halys venom platelet inhibitor (AHV-PI) group(18 rats). The AHV-PI experimental group was divided into low, middle and high dose groups according to the dose of AHV-PI injected into sublingual vein (0.05, 0.1, 0.2 mg/kg), with 6 rats in each group. Electrocardiogram(ECG) changes of rats were monitored by RM6240 biological signal collection and processing system. Western blot was used to monitor the expression of platelet membrane glycoprotein VI (GPVI) in rats under the intervention of AHV-PI. Blood coagulation time (R), blood clots forming time (K), Alpha Angle (A), and maximum amplitude of blood coagulation (MA) were assayed by Thrombelastography (TEG5000).Platelet aggregation rate was measured by turbidimetry. RESULTS:Compared with MIRI model group, the expression level of GPVI AHV-PI medium dose experimental group and high dose experimental group were significantly decreased (P<0.05), and the R and K values were significantly prolonged, while the A value and MA values were significantly decreased (P<0.05). Platelet aggregation time, aggregation amplitude and aggregation rate were significantly decreased (P<0.05). CONCLUSION: AHV-PI can significantly inhibit the expression of GPVI in MIRI rats and improve the hemorheological properties related to myocardial injury in rats, thus weakening the injury process.
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AIM: To explore the effect of inhibiting proliferation and inducing apoptosis in human primary gastric tumor by the Agkis-trodon halys venom anti-tumor component (AHVAC-). METHODS: Human primary gastric cancer cells were isolated by trypsin digestion, serum-free culture, and purified by differential adherence method, and cells were identified by immunohistochemistry. Cell proliferation and toxicity assay (CCK-8) was used to detect the inhibition rate of AHVAC- in different concentrations of primary gastric cancer cells. Immunohistochemistry was used to verify the apoptosis of human primary gastric cancer cells induced by AHVAC- and the morphological changes were observed by Hematoxylin-Eosin staining (HE staining). AHVAC--induced primary gastric cancer cell transformation rate was detected by flow cytometry Annexin V/PI double staining.RESULTS: Seven human primary gastric cancer cells were successfully isolated and purified, and 11 cases failed. Immunohistochemical identifications of carcinoembryonic antigen (CEA) and broad-spectrum keratin protein (AE1/AE3) were positive for both antibodies. AHVAC- inhibited the proliferation of human primary gastric cancer cells and showed a dose-dependent effect (P<0.01). Immunohistochemistry showed that the expression level of cysteine aspartic protease-3 (Caspase-3) up-regulated with the increase of AHVAC- concentration. HE staining showed that with the increase of AHVAC- concentration, the cell gap increased, nuclear pyknosis, and apoptosis cells increased. Flow cytometry showed that the apoptosis rate of human primary gastric cancer cells up-regulated with the increase of AHVAC- concentration (P<0.05). CONCLUSION: AHVAC- can inhibit the proliferation and induce apoptosis in human primary gastric cancer cells in a dose-dependent manner.
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Agkistrodon acutus is a traditional Chinese herb medicine which has immunological regulation,anti-tumor,anti-inflammatory and analgesic effects,which is mainly used for the treatment of rheumatoid arthritis,ankylosing spondylitis,sjogren's syndrome and tumors. In order to excavate more important functional genes from A. acutus,the transcriptome of the venom gland was sequenced by the Illumina Hi Seq 4000,and 32 862 unigenes were assembled. Among them,26 589 unigenes were mapped to least one public database. 2 695 unigenes were annotated and assigned to 62 TF families,and 5 920 SSR loci were identified. The majority of mapped unigenes was from Protobothrops mucrosquamatus in the NR database,which revealed their closest homology. Three secretory phospholipase A_2 with different amino acid sequences showed similar spatial structures and all had well-conserved active sites. The 3 D structural models of C-type lectin showed conserved glycosylation binding sites( Asn45). This study will lay the foundation for the further study of the function of snake venom protein,and promoting the development and utilization of genome resources from A. acutus.
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Animals , Agkistrodon/genetics , Crotalid Venoms , Gene Expression Profiling , Snake Venoms/genetics , Snakes , TranscriptomeABSTRACT
AIM:To explore the role of nuclear factor-κB(NF-κB)and activator protein-1(AP-1)signaling pathway in the inhibitory effects of Agkistrodon acutivirus protein C activator(PCA)on lipopolysaccharide(LPS)-induced tissue factor(TF)expression in human umbilical vein endothelial cells(HUVECs).METHODS: The viability of the HUVECs was measured by MTT assay.The protein distribution of tumor necrosis factor-associated factor 6(TRAF6)in the cells was detected by immunohistochemical staining.The protein expression of NF-κB p65,TF,c-Fos and c-Jun was deter-mined by Western blot.The mRNA expression of TF in the HUVECs was detected by qPCR.The content of TF in the me-dium of each group was measured by ELISA.RESULTS:Compared with the control group,the viability of the HUVECs in LPS group decreased significantly(P<0.01), obvious yellow dye particles appeared in the cytoplasm, cytoplasmic stai-ning deepened,and the average absorbance of TRAF6 was increased(P<0.01).The protein expression of NF-κB p65, c-Jun and c-Fos were significantly increased(P<0.01).The expression of TF at mRNA and protein levels were signifi-cantly increased(P<0.01).Compared with the LPS group,the cell viability in PCA +LPS group was slightly increased (P<0.05),the cell morphology was normal,cytoplasmic yellow dye particles were not obvious, and the average absor-bance of TRAF6 was significantly lower than that in LPS group(P<0.01).The protein expression of NF-κB, c-Jun and c-Fos was significantly decreased(P<0.01),and the expression of TF at mRNA and protein levels were decreased(P<0.01).CONCLUSION:PCA significantly reduces the damage of HUVECs induced by LPS.The mechanism may be a-chieved by reducing the activation of TRAF 6,NF-κB and AP-1 nuclear transcription factors,thereby reducing the release of tissue factor.
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Objective To investigate the renal protective effect of antivenom therapy assisted with hyperbaric oxygen(HBO) in agkistrodon acutus poisoning rat.Methods Ninety-six healthy adult male SpragueDawley rats were randomized into the control group(group A),snake venom poisoning group(group B),antivenom treatment group(group C) and antivenom+ HBO treatment group(group D),24 cases in each group.The group B,C and D were injected with agkistrodon acutus toxin 0.8 × LD50 (LD50 =1.594 mg/kg) by tail vein for establishing the poisoning model.The renal tissue pathological changes after agkistrodon acutus poisoning were observed.In the antivenom treatment group,antivenom(0.8 ×＜ 78 U/kg) was injected by tail vein after successfully constructing model,the group D was given once HBO treatment at 0,4,11 23 h after antivenom injection,while the group A was not given any treatment.Results The rats appeared obvious poisoning symptoms after snake venom intravenous injection.Compared with the group B,therat renal function and coagulation functions in the group C and D were significant improved,more over the group D was better than the group C(P＜0.05).The rats of the group B appeared glomerular hemorrhage,renal capsule expansion,different degrees of congestion in peripheral blood capillaries,renal tubular cavity dilation,cellular fatness,falling of brush border,vacuolation of renal tubular epithelial cells and inflammary cells infiltration in renal interstitial tissue.Along with time lapse,the renal hemorrhage was decreased and inflammatory cells infiltration was increased.The abovelesions in the group C and D were significantly improved.Conclusion The Agkistrodon acutus poisoning can lead to renal damage in rats;the antivenom treatment assisted with HBO has the protective effect on renal injury in agkistrodon acutus poisoning rat,moreover early adopting has better protective effect.
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El presente trabajo muestra una experimentación pura basada en la preparación homeopática del veneno de la serpiente Agkistrodon bilineatus, algo que ha demostrado su eficacia en diferentes aplicaciones clínicas que datan de la época de Constantine Hering, uno de los pioneros de la Homeopatía en Estados Unidos. Los resultados son alentadores, toda vez que los órganos con mayor número y diversidad de síntomas fueron el sistema nervioso central, los ojos, así como los aparatos respiratorio y digestivo. En conclusión, este derivado del veneno de la serpiente en referencia, preparado homeopáticamente, puede emplearse debido a que se generó una patogenesia concreta en un grupo de experimentadores.
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Humans , Male , Female , Materia Medica , Hahnemannian Method , Homeopathic Semiology , Agkistrodon , Clinical ClerkshipABSTRACT
Objective:To establish a method for the separation and purification of haemocoagulase from agkistrodon blomhoffii in Changbai Mountain. Methods: An enzyme component with clotting activity was isolated from the venin of agkistrodon blomhoffii in Changbai Mountain by gel filtration, anion-exchange chromatography and heparin-Sepharose affinity chromatography. SDS-Page and RP-HPLC were used to determine its purity, SDS-Page was applied to study its action mode with bovine fibrinogen, HPSEC was used to determine the molecular weight, an IEF method was employed to detect its isoelectric point, and Lowry method was used for the deter-mination of protein concentration. Results:One haemocoagulase was purified from agkistrodon in Changbai Mountain. SDS-Page dis-played one band, and HPLC showed one single chromatographic peak. The haemocoagulase acted only on α chain of fibrinogen. Its molecular weight was 32. 2kD with isoelectric point of 5. 21. The enzyme had clotting activity in vitro. Conclusion:The method can be used for the separation and purification of haemocoagulase from agkistrodon blomhoffii in Changbai Mountain.
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OBJECTIVE:To optimize the best crushing particle size of Agkistrodon decocting powder,and study its effect on apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes. METHODS:Pre-column derivatization RP-HPLC was adopted. Using the total decoction amounts of 4 main amino acids (aspartic acid,glutamic acid,L-hydroxyproline,glycine) that decocted once as index,the Agkistrodon decoction pieces and those through No.1-8 seive were screened,and the best crushing particle size of Agkistrodon decocting powder was optimized. The human rheumatoid arthritis fibroblast-like synoviocytes were divided into nega-tive control group,positive control group (1 μmol/L methotrexate) and the best crushing particle size of Agkistrodon decocting powder group(2.0 mg/mL),flow cytometry was used to determine the cell apoptosis after cultured for 48 h. RESULTS:The best crushing particle size was through No.6 seive,when the total amount of 4 main amino acids was(61.27±0.02)mg/g(n=3). Com-pared with negative control group,the apoptosis rate in the best crushing particle size of Agkistrodon decocting powder group was significantly increased(P<0.05),which was slightly higher than positive control group. CONCLUSIONS:The best crushing parti-cle size is through No.6 seive;and Agkistrodon decocting powder shows effect on reducing the apoptosis of human rheumatoid ar-thritis fibroblast-like synoviocytes.
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OBJECTIVE:To optimize the best crushing particle size of Agkistrodon decocting powder,and study its effect on apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes. METHODS:Pre-column derivatization RP-HPLC was adopted. Using the total decoction amounts of 4 main amino acids (aspartic acid,glutamic acid,L-hydroxyproline,glycine) that decocted once as index,the Agkistrodon decoction pieces and those through No.1-8 seive were screened,and the best crushing particle size of Agkistrodon decocting powder was optimized. The human rheumatoid arthritis fibroblast-like synoviocytes were divided into nega-tive control group,positive control group (1 μmol/L methotrexate) and the best crushing particle size of Agkistrodon decocting powder group(2.0 mg/mL),flow cytometry was used to determine the cell apoptosis after cultured for 48 h. RESULTS:The best crushing particle size was through No.6 seive,when the total amount of 4 main amino acids was(61.27±0.02)mg/g(n=3). Com-pared with negative control group,the apoptosis rate in the best crushing particle size of Agkistrodon decocting powder group was significantly increased(P<0.05),which was slightly higher than positive control group. CONCLUSIONS:The best crushing parti-cle size is through No.6 seive;and Agkistrodon decocting powder shows effect on reducing the apoptosis of human rheumatoid ar-thritis fibroblast-like synoviocytes.
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Objective To investigate the mechanism of platelet inhibitor from Agkistrodon halys venom (AHV-PI) on platelet aggregation. Methods Protein kinase Akt phosphorylation levels in platelet were measured by Western blot. XS-1000I blood cell counter was used for platelet count. The plasma content of 5'-NT and platelet membrane GPIb were determined by Enzyme-Linked Immunosorbnent Assay (ELISA). The effect of AHV-PI on binding rate between the fluorescence labeled monoclonal antibody CD61 (FITC-CD61) and platelet membrae glycoprotein Ⅱb/Ⅲa (GPⅡb/Ⅲa) was observed by flow cytometry (FCM). Results AHV-PI can reduce the level of Akt-phosphorylation level and the number of platelet. AHV-PI can increase the content of 5'-NT in plasma, reduce the expression of platelet GPIb. Flow cytometry displayed AHV-PI can not affect the rate of combination between platelet GPⅡb/Ⅲa and FITC-CD61. Conclusion The mechanism of inhibition of platelet aggregation may be inhibit protein kinase Akt phosphorylation to block the signal transduction pathway of Akt. Limit cell grouth and reduce platelet number, also it may be related to its 5'-NT activity, it can degradate ADP to prevent the formation of platelet thrombus.
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Objective:In order to look for a good method for preparation of hemorrhagin antiserum. Methods: Three kinds of hemorrhagins including AaH Ⅰ, AaH Ⅱ, and AaH Ⅳ were purified from Agkistrodon acutus venom according to predecessors's methods and crude AaH Ⅰ, AaH Ⅱ and AaH Ⅳ were obtained. Preparation electrophoresis was used to purify AaH Ⅰ,AaH Ⅱand AaH Ⅳ further. As for an hemorrhagin, six different dyeing methods were used to dye PAGE gel and the gel contained hemorrhagin was obtained respectively. The ground gel contained hemorrhagin was used to immune mice and its antiserum was obtained. Antiserums quality was tested through ELISA test and neutralization of the hemorrhagic activities of corresponding hemorrhagin. Results:Effective IgG concentration in different antiserum was different and effective IgG made through non toxic type protein fast stain reagent kit was higher than others. Conclusion:Non toxic type protein fast stain reagent kit is the best dyeing method among the six dyeing methods.
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OBJECTIVE:To guide rational use of Heamocoagulase agkistrodon for injection,and to provide reference for phar-maceutical care. METHODS:Treatment methods and outcome of 9 cases of allergic shock induced by Heamocoagulase agkistrodon for injection were retrieved from CNKI and VIP database during 2009-2015,and then analyzed statistically,based on one case of allergic shock induced by Heamocoagulase agkistrodon for injection in the Affiliated Hospital of Zhunyi Medical College. RE-SULTS:10 pationts with allergic shock induced by Heamocoagulase agkistrodon for injection mainly used the drug for hemostasis before and after surgery,heavy menstrual bleeding,etc.,with dose of 1 or 2 U. Route of administration included intravenous injec-tion and intravenous dripping. Allergic shock often occurred within 30 min after medication. CONCLUSIONS:The use of Heamoco-agulase agkistrodon for injection in some patients belongs to off-label drug use,and route of administration is different from that stated in package inserts. Allergic shock induced by Heamocoagulase agkistrodon for injection occurs rapidly;the drug should be used strictly according to package inserts and authoritative literatures;emergency plan should be developed before medication in or-der to guarantee the safety of drug use.
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<p><b>OBJECTIVE</b>To determine the effective dosage and formulation of agkistrodon in collagen-induced arthritis (CIA) rats.</p><p><b>METHODS</b>CIA was induced by injection of collagen in complete/incomplete Freund's adjuvant. Agkistrodon decoction, agkistrodon powder, and agkistrodon wine were administered daily starting from the onset of arthritis. Paw swelling degree was measured by using a volume-measuring instrument every 7 days after primary immunization. Arthritis index was measured and calculated using the "five scoring method" every 7 days. The levels of serum interleukin-1ß (IL-1ß) and type II collagen IgG antibodies were detected by enzyme-linked immunosorbent assay. Finally, all ankles were removed, and X-ray radiography was performed with In-vivo Imaging System FX. Samples were counterstained with hematoxylin and eosin for analysis.</p><p><b>RESULTS</b>Among the various dosage formulations of agkistrodon, high-dose powder, which was equivalent to an amount of 6 g/day in adults, showed better effects on the inhibition of joint swelling and reduction of arthritis index score. The relatively low levels of serum IL-1 and anti-type II collagen IgG antibodies, as well as the X-ray radiography and pathology results, further proved the superiority of the high-dose powder over the other formulations. The effect of decoction on inhibiting joint swelling was inversely proportional to the dosage. Other effects, such as reduction of arthritis index score and the levels of serum IL-1 and anti-type II collagen IgG antibodies, were directly proportional to the dosage. While the use of large dose agkistrodon wine led to negative effects.</p><p><b>CONCLUSION</b>These data highlight the potential function of high-dose agkistrodon powder, which was equivalent to an amount of 6 g/day in adults. The powder can quickly relieve the symptoms of rheumatoid arthritis and prevent aggravation of disease, especially during the early period.</p>
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Animals , Female , Agkistrodon , Metabolism , Antibodies , Blood , Arthritis, Experimental , Blood , Drug Therapy , Collagen Type II , Allergy and Immunology , Dosage Forms , Dose-Response Relationship, Drug , Enzyme-Linked Immunosorbent Assay , Extremities , Diagnostic Imaging , Pathology , Interleukin-1beta , Blood , Medicine, Chinese Traditional , Rats, WistarABSTRACT
Objective To systematically evaluate the efficacy and safety of injected hemocoagulase Agkistrodon for intraoperative bleeding.Methods We electronically searched several Chinese database till May 2014 to identify randomized controlled trials (RCTs) about injected hemocoagulase Agkistrodon for operative hemorrhage.The methodological quality of included RCTs was assessed,and the data were extracted by two reviewers independently according to the Cochrane Handbook.The homogeneous RCTs were pooled using RevMan software (Version 5.1.0).Results 2 358 articles were searched initially,and 12 RCTs involving 1 031 patients met the inclusion criteria.The results of Meta-analyses showed that:in the study groups,the average haemostatic time of the wound is shorter [OR =-40.29,95％ CI(-71.13,-9.44),P =0.01],the hemorrhagic volume of the wound [OR =-1.38,95％ CI(-1.90,-0.87),P ＜ 0.000 01],hemorrhagic volume per unit area of the wound [OR =-0.05,95％ CI(-0.06,-0.03),P ＜ 0.00001],introperative hemorrhagic volume and drainage volume on POD1 [OR =-18.52,95％ CI (-25.50,-11.53),P ＜ 0.000 01] are lower than the placebo groups.In all studies,no difference was found in the variation of hemostatic function,liver and renal function and drug-induced adverse events.Conclusions The injection of hemocoagulase Agkistrodon can significantly reduce the average haemostatic time of the wound,the hemorrhagic volume of the wound,overall introperative hemorrhagic volume and drainage volume on POD1,and does not affect the hemostatic function,liver and renal function and not increase the incidence of adverse effects or postoperative complications.
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Objective To investigate the efficiency and safety of agkistrodon hemocoagulase on coagulation function in pa?tients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB). Methods In this prospective,randomized controlled trial,80 eligible patients accepted valve replacement were assigned to control group (n=20) and agkistrodon hemoco?agulase groups (H1, H2, H3) according to the different timing of administration. Twenty patients were given treatment 20 minutes before anesthesia induction (H1 group), 20 patients were given treatment 20 minutes after CBP (H2 group) and 20 patients were given treatment after CBP (H3 group). Coagulation parameters including prothrombin time (PT), activated partial thromboplas?tin time (APTT), thrombin time (TT), fibrinogen (Fib) and platelet (PLT) were detected 20 minutes before surgery (T0), immedi?ately after surgery (T1) and 24 hours after surgery(T2). Data of 24-h postoperative drainage of mediastinal and pericardial, dura?tion of mechanical ventilation, stay time of intensive care unit (ICU), the actual days of hospitalization and hospital costs were recorded. The clinical parameters were also recorded including blood transfusion after surgery, secondary thoracotomy, aller?gies, liver and kidney dysfunction, deep vein thrombosis and neuropsychiatric symptoms. Results Compared with control group, values of PT, APTT and TT at T1 and T2 were significantly lower in H1, H2 and H3 groups (P<0.05). Compared with T0, values of PT, APTT and TT at T1 and T2 were significantly higher in all groups of patients (P<0.05). Values of PT and TT at T1 and T2 were significantly decreased in H3 group than those of H1 group (P<0.05). The pericardial and mediastinal drainage, the duration of ventilation support within 24-h after surgery were significantly lower in H1, H2 and H3 groups than those of control group (P<0.05). There was no significant difference in the incidence of adverse events between groups. Conclusion Agkis?trodon hemocoagulase is safe and effective in patients undergoing cardiac valve replacement with cardiopulmonary bypass.
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OBJECTIVE:To optimize the decoction process of agkistrodon. METHODS:RP-HPLC with pre-column derivatiza-tion was adopted. With the contents of 4 main amino acids in agkistrodon as index,the decoction process(decocting times,heating time,water quantity and medicinal material crushing granularity) was optimized by orthogonal tests and verified. RESULTS:The optimal technology of the decoction process of agkistrodon was as 3 times of decoction,60 min of heating time,50 ml of water consumption for 0.90 g medicinal material and No.6 sieve. The results of verification test showed the total extraction of 4 kinds of amino acids was 72.68 mg/g(RSD=3.77%,n=3). CONCLUSIONS:The decoction technology can be used for the decoction pro-cess of agkistrodon,and it is stable and feasible.