Mamushi is a species of pit viper distributed throughout Japan excluling the Ryukyu Islands and sighted from spring to autumn. It is estimated that about ten people dies in a year. It is important in the rural medicine because a lot of people are bitten in mountains and fields. A total of 35 cases of mamushi viper bite were treated from 1999 to 2006 in our hospital. The patients were 17 men and 18 women ranging in age from 7 to 80 years old (average: 60). Local swelling and pain were manifest in all the cases. Sixteen patients had systemic symptoms, and toxic effects commonly appeared in the eyes. The blood tests revealed elevated CPK levels in 24 patients (69%). The elevation correlated to the seriousness of the clinical symptoms. In accordance with our manual for mamushi viper bites, all patients were hospitalized after incision for exclusion of the toxin. The median length of time before the worst symptoms began to subside was 3 days. The median length of hospitalization stays was 7 days. It took long before the patients fully recovered. The severity of envenomation was different from patient to patient and one case needed intensive care. The median of treatment period was 31 days. The quick and appropriate primary care for the mamushi viper bits is important to prevent serious complications.
Median Statistical Measurement ; symptoms <1> ; Agkistrodon halys blomhoffi ; Clinical ; Cases

BACKGROUND AND PURPOSE: Arginine esterase(Ancrod), a thrombin-like enzyme, purified from the venoms of Agkistrodon halys, has known to cleave fibrinopeptide A from the fibrinogen and lead to circulation of soluble noncross-linked "ancrodfibrin', which stimulates endogenous T-PA release.Many authors have suggested clinical applicability of this enzyme,but clinical studies on its fibrinolytic action has been insufficient.Thus we studied the influence of this enzyme on fibrinolytic activity in cerebral infarction. METHOD: We observed the change of euglobulin fibrinolytic activity, t-PA antigen, t-PA activity, fibrinopeptide A, fibrinogen, FDP and D-dimer, during 12 hours after a bolus intravenous administration of 0.25 unit of the arginine esterase to the 9 patients with cerebral infarction. RESULT:There was no change of the euglobulin fibrinolytic activity, fibrinopeptide A and t-PA Ag but there was significant increase in both t-PA activity and FDP, D-dimer and significant decrease in fibrinogen. CONCLUSION: Our result suggest that arginine esterase converts fibrinogen to a fibrin polymer which has a increased susceptibility to lysis by plasmirl This enzyme seems to amplify T-PA activity through the consequent increase in FT)P, because there is no increase in the euglobulin fibrinolytic activity, fibr'mopeptide A and t-PA Ag suggesting direct T-PA release. Arginine esterase, having action of effective defibrinogenation and safe fibrinolysis,could be used for the thrombus related disease.
Administration, Intravenous ; Agkistrodon ; Arginine ; Cerebral Infarction ; Fibrin ; Fibrinogen ; Fibrinopeptide A ; Humans ; Polymers ; Snake Venoms* ; Snakes* ; Thrombosis ; Venoms

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.
Agkistrodon/genetics* ; Animals ; Crotalid Venoms ; Gene Expression Profiling ; Snake Venoms/genetics* ; Snakes ; Transcriptome

Venomous snakes are estimated to inflict 400,000 bites annually, resulting in approximately 40,000 deaths. There are nearly 3,500 known species of snakes worldwide, and three species exist in Korea. Venom contains toxins that effect the cardiovascular system, the kidneys, the respiratory system and the muscles. There are many modalities in treating snake bites, but most of all, neutralization of the venom is the most important. however, many doctors in Korea hesitate in using antivenom. The reason is that there are no specific antivenoms against Korean venous snakes. In this case, a 2-year-old female child with a snake bite vistied our hospital Korea via a local clinic. Severe systemic symptoms and local symptoms, including compartment syndrome, were observed. When we tried antivenom, the result was satisfactory.
Agkistrodon* ; Antivenins ; Cardiovascular System ; Child ; Child, Preschool ; Compartment Syndromes ; Female ; Humans ; Kidney ; Korea ; Muscles ; Respiratory System ; Snake Bites* ; Snakes* ; Venoms

Thrombin-like enzymes (TLEs) are studied widely because of their therapeutic potential in myocardial infarction and thrombotic diseases. We synthesized the DNA fragment encoding thrombin-like enzyme calobin from Agkistrodon caliginosus (Korean Viper) venom by fusion PCR and expressed it in Pichia pastoris. After induction by 0.5% methanol for 48 h, the expression level of recombinant calobin reached 3.5 g/L in medium. The recombinant calobin was purified by Q-Sepharose Fast Flow ion-exchange chromatography and Sephacryl-S-100 gel filtration chromatography. Purified sample had a molecular weight of 32 kD shown in SDS-PAGE. It hydrolyzed fibrinogen and formed a light white hydrolysis circle in fibrinogen plate. SDS-PAGE analysis showed that recombinant calobin cleaved Aalpha-chain of fibrinogen specifically, and produced an appropriately 40 kD new band. However, we failed to find its fibrin-clot formation activity.
Agkistrodon ; Animals ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Serine Endopeptidases ; biosynthesis ; genetics ; Thrombin ; biosynthesis ; genetics ; Viper Venoms ; enzymology

Human sparganosis is a zoonotic disease caused by infection with larval forms (procercoid/plerocercoid) of Spirometra spp. The purpose of this study was to identify Spirometra spp. of infected snakes using a multiplex PCR assay and phylogenetic analysis of mitochondrial DNA sequence data from the spargana of terrestrial snakes obtained from Korea and China. A total of 283 snakes were obtained that included 4 species of Colubridae comprising Rhabdophis tigrinus tigrinus (n=150), Dinodon rufozonatum rufozonatum (n=64), Elaphe davidi (n=2), and Elaphe schrenkii (n=7), and 1 species of Viperidae, Agkistrodon saxatilis (n=60). The snakes were collected from the provinces of Chungbuk, Chungnam, and Gyeongbuk in Korea (n=161), and from China (n=122). The overall infection rate with spargana was 83% (235/283). The highest was recorded for D. rufozonatum rufozonatum (100%), followed by A. saxatilis (85%) and R. tigrinus tigrinus (80%), with a negative result for E. davidi (0%) and E. schrenkii (0%). The sequence identities between the spargana from snakes (n=50) and Spirometra erinaceieuropaei (KJ599680) or S. decipiens (KJ599679) control specimens were 90.8% and 99.2%, respectively. Pairwise genetic distances between spargana (n=50) and S. decipiens ranged from 0.0080 to 0.0107, while those between spargana and S. erinaceieuropaei ranged from 0.1070 to 0.1096. In this study, all of the 904 spargana analyzed were identified as S. decipiens either by a multiplex PCR assay (n=854) or mitochondrial cox1 sequence analysis (n=50).
Agkistrodon ; China* ; Chungcheongbuk-do ; Chungcheongnam-do ; Colubridae ; DNA, Mitochondrial ; Gyeongsangbuk-do ; Humans ; Korea* ; Multiplex Polymerase Chain Reaction ; Sequence Analysis ; Snakes* ; Sparganosis ; Sparganum ; Spirometra* ; Viperidae ; Zoonoses

This study was carried out to confirm the infection source of the human case of Fibricola seoulensis, and to reveal out a part of its life cycle in Korea. Also the morphological characteristics of the metacercaria were described. The results were summarized as follows: Rana nigromaculata and Natrix tigrina lateralis were found to be infected naturally by the metacercariae(diplostomula) of F. seoulensis. The metacercarial capsule was round to elliptical in tissue of the intermediate hosts with a long diameter 0.232 - 0.385 mm. Liberated metacercariae were ovoid with small conical posterior body. Body length measured 0.199 - 0.312 mm and width 0.153 - 0.252 mm. The infection rate of R. nigromaculata by the metacercariae ranged from 87% to 100% by area, and the number of the larvae ranged from 3 to 390 by frog. The metacercariae were found in skeletal muscle of frogs, from head to hindlegs. All examined N. t. lateralis were found to be infected by the metacercariae with the range of numbers frome 3 to 35,918. The larvae were collected from all viscera and body segments except for the head of the snakes. However, a great majority of the metacercariae were collected from the stomach. Hematoxylin-eosin stained preparations of frog skeletal muscle and snakes stomach revealed that the metacercariae had no cyst wall of worm origin, but encapsulated by the host tissue. Some of them were found in dilated lymphatic vessels. The larval infection was associated with slight or severe inflammatory reaction even with granuloma formation. By above results, it was concluded that the frog, Rana nigromaculata, was the second intermediate host, and the snake Natrix tigrina lateralis was a paratenic host of F. seoulensis in nature in Korea.
parasitology-helminth-trematoda ; Fibricola seoulensis ; metacercaria ; host ; frog ; Rana nigromaculata ; Rana rugosa ; Natrix tigrina lateralis ; Elaphae dione ; Dinodon rufozonatum rufozonatum ; Agkistrodon blomhoffi brevicaudus ; epidemiology

A novel serine protease with high purity was extracted from the venom of Agkistrodon hlays Pallas using monoclonal antibody affinity chromatography. This protease releases bradykinin and has arginine esterase activity without being activated. After purification, its hydrolytic activity exceeded 800 U/mg, far higher than its counterparts from mammalian sources. The purity of the kininogenase could exceed 95%. The acute toxicity and the long-term toxicity of this kallikrein was studied for its potential clinical application. The maximum tolerance dose in adult was 150,000 times greater than the maximum applied dose, and long-term administration at the dose 50 times of allowed clinical dose did not obviously after the animals' body weight, survival condition, liver function, renal function, or blood routines, suggesting the extremely low toxicity of the kallidrein.
Agkistrodon ; Animals ; Bradykinin ; metabolism ; Crotalid Venoms ; toxicity ; Kallikreins ; toxicity ; Maximum Tolerated Dose ; Mice ; Mice, Inbred BALB C ; Serine Endopeptidases ; toxicity ; Toxicity Tests, Acute

The study was purposed to investigate the effect of extract of Agkistrodon Halys venom on proliferation and apoptosis of K562 cells. The inhibition of K562 cell proliferation was measured by MTT assay; The morphologic changes of K562 cells was observed by microscopy; the apoptosis of K562 cells was measured by flow cytometry; the activity of extracellular signal-regulated kinase (ERK) in K562 cells was detected by Western blot. The results showed that when K562 cells were treated with 0, 1, 10, 20 microg/ml of the extraction for 48 hours, the apoptosis rates were 2.1%, 21.3%, 49.7%, 70.1%, respectively. The proliferation of K562 cells was obviously inhibited in dose-dependent manner. Typical morphologic changes significantly appeared in the extract-treated K562 cells. The extract obviously inhibited the activity of ERK in K562 cells. It is concluded that the extract of Agkistrodon Halys' venom can inhibit the proliferation of K562 cells and induce apoptosis of K562 cells.
Agkistrodon ; Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Complex Mixtures ; pharmacology ; Crotalid Venoms ; chemistry ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; K562 Cells

BACKGROUNDSnake venom contains a number of components with different pharmacological and biological activities, especially in cancer therapy, and has increasingly become a research focus. This study was designed to isolate and purify a novel anti-clotting protein component from the venom of Agkistrodon acutus, and to explore its physico-chemical properties and biological activity.

METHODSThe venom of Agkistrodon was isolated and purified by ion-exchange chromatography on diethylaminoethyl (DEAE)-Sepharose Fast Flow, molecular sieve filtration through Sephadex G75, SP-Sepharose Fast Flow and molecular sieve filtration through Sephadex G50. We detected the activated partial thromboplastin time (APTT) of the eluant to select the anti-clotting protein component of interest. The molecular weight was determined by sodium dodecyl sulfate-polyacrylamid gel electrphoresis (SDS-PAGE) and liquid chromatography. Its protein content was detected by bicinchoninic acid (BCA).

RESULTSSDS-PAGE vertical gel electrophoresis showed that the anticoagulant factor is a tripolymer composed of three proteins whose molecular weights are 25 KDa, 30 KDa and 50 KDa. The factor contains about 65% percent protein.

CONCLUSIONSA novel anti-clotting protein component was purified by ion-exchange chromatography and molecular sieve filtration from the venom of Agkistrodon acutus and was found to be composed of three kinds of proteins.

Agkistrodon ; metabolism ; Animals ; Anticoagulants ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Crotalid Venoms ; chemistry ; Electrophoresis, Polyacrylamide Gel ; Proteins ; chemistry ; isolation & purification