OBJECTIVE: To report the prevalence of lupus anticoagulants and anticardiolipin antibodies in patients with recurrent spontaneous abortion and infertility. MATERIAL AND METHOD: Lupus anticoagulants and anticardiolipin antibodies were analyzed by Diluted Russell's Viper Venom Test (DRVVT) and solid phase enzyme immunoassay, respectively. RESULTS: In 200 patients with infertility, there were 6 cases (3%) with positive lupus anticoagulants or anticardiolipin antibodies. Of these, 3 patients (1.5%) showed positive lupus anticoagulants and anticardiolipin antibodies, respectively. In 120 patients with recurrent spontaneous abortion, there were 13 cases (10.8%) of positive lupus anticoagulants or anticardiolipin antibodies. Of these, one patient (1%) showed lupus anticoagulants and 12 patients (10%) showed anticardiolipin antibodies. But in two groups, there was no cases with positive lupus anticoagulants and anticardiolipin antibodies. CONCLUSION: Lupus anticoagulants and anticardiolipin antibodies are definite cause of recurrent spontaneous abortion. There has been a speculation that they might be associated with infertility and repeated IVF failures. But it was found that the role of lupus anticoagulants and anticardiolipin antibodies in these cases are not clear.
Abortion, Spontaneous* ; Antibodies, Anticardiolipin* ; Anticoagulants* ; Female ; Humans ; Immunoenzyme Techniques ; Infertility* ; Pregnancy ; Prevalence ; Russell's Viper ; 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

The present study evaluated the effects of Androctonus amoreuxi scorpion venom, Cerastes cerastes snake venom and their mixture on prostate cancer cells (PC3). An MTT assay was used to determine the anti-proliferative effect of the venoms, while quantitative real time PCR was used to evaluate the expression of apoptosis-related genes (Bax and Bcl-2). Furthermore, colorimetric assays were used to measure the levels of malondialdehyde (MDA) and antioxidant enzymes. Our results show that the venoms significantly reduced PC3 cell viability in a dose-dependent manner. On the other hand, these venoms significantly decreased Bcl-2 gene expression. Additionally, C. cerastes venom significantly reduced Bax gene expression, while A. amoreuxi venom and a mixture of A. amoreuxi & C. cerastes venoms did not alter Bax expression. Consequently, these venoms significantly increased the Bax/Bcl-2 ratio and the oxidative stress biomarker MDA. Furthermore, these venoms also increased the activity levels of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Overall, the venoms have cytotoxic and anti-proliferative effects on PC3 cells.
Apoptosis ; Catalase ; Cell Survival ; Gene Expression ; Genes, bcl-2 ; Glutathione Peroxidase ; Glutathione Reductase ; Hand ; Humans* ; Malondialdehyde ; Oxidative Stress ; Prostate* ; Prostatic Neoplasms* ; Real-Time Polymerase Chain Reaction ; Scorpion Venoms ; Scorpions* ; Snake Venoms* ; Snakes* ; Superoxide Dismutase ; Venoms ; Viper Venoms ; Viperidae*

The metalloproteinases/disintegrins in the snake venom act as platelet aggregation inhibitor by an antagonism against integrin on platelet through its RGD sequence and may play other important role in cell-cell fusion, cell matrix interaction and other cellular function. Ussurin is a new metalloproteinase/disintegrin that was cloned from Gloydius ussuriensis. Poly (A+) RNA was purified from the total RNA preparation from venom gland of a single G. ussuriensis using the poly (A+) tract-mRNA isolation system. A cDNA library was constructed with the SMART PCR cDNA library construction kit. The cDNA library was screened and the positive clones were selected. The full-length cDNA of Ussurin was obtained. The cDNA encoding the Ussurin precursor has a 51bp 5'-UTR, the open reading frame of Ussurin and a 490 bp 3'-UTR, the open reading frame of Ussurin cDNA nucleotide sequence is 1434 bp and codes for 478 amino acids with a predicted molecular mass of 53.2 kD and an isoelectric point of 5.37. There is no potential N-glycosylation site in the deduced sequence region. Its deduced amino acid sequence consists of four region, a signal sequence of 18 amino acid residues, a zymogen pro-peptide of 171 amino acid residues with a cysteine switch motif (PK-MCGVT) in it, a central metalloproteinase domain of 201 amino acid residues containing a conserved zinc-chelating sequence (HEXXHXXGXXH) and a methionine-turn CIM involving zinc banding also, a space sequence between metalloproteinase domain and disintegrin domain of 15 amino acid residues with a conserved T392, T397, S400, which is specific residues of the P-II snake venom metalloproteinases, a disintegrin domain of 73 amino acid residues with a characteristic RGD region and six-disulfide bonds. Ussurin belongs to P-II class. The cDNA sequence and deduced amino acid sequence of Ussurin precursor were compared with homologous sequence in the GenBank database, the result reveals high degree of homology in sequence and organization pattern of domain with metalloproteinase/disintegrin gene family of other snake species. Compared with the alignment of amino acid sequence of metalloproteinase/disintegrin member, hypervariable regions of this member were revealed, besides they share higher homologous in the zymogen domain. It suggests that the hypervariable regions are the counterparts directly suitable for interacting with different domain of receptors, different receptors or substrates.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Disintegrins ; chemistry ; genetics ; metabolism ; Metalloproteases ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Sequence Alignment ; Viper Venoms ; enzymology ; genetics ; Viperidae ; genetics ; metabolism

This study was aimed to investigate the hemostatic effects of hemocoagulase agkistrodon (HCA) and its mechanism. The procoagulative and hemostatic effects of HCA were evaluated by using rabbit blood coagulatin time and mouse tail bleeding time; the mechanisms of HCA hemostatic effect were analyzed by using rabbit blood clot lysis and fibrinogen lysis. The results showed that HCA shortened the rabbit blood coagulation time and the mouse tail bleeding time significantly. The effects are nearly similar to that of positive control (reptilase). HCA also induced rabbit blood clot lysis and directly hydrolysed the alpha-chain of fibrinogen. It is concluded that HCA exert its hemostatic effects by hydrolysing the alpha-chain of fibrinogen, but it is not able to induce production of XIII factor.
Agkistrodon ; Animals ; Batroxobin ; isolation & purification ; pharmacology ; Bleeding Time ; Blood Coagulation ; drug effects ; Crotalid Venoms ; enzymology ; Fibrinogen ; metabolism ; Hemostatics ; pharmacology ; Mice ; Rabbits ; Random Allocation

OBJECTIVETo investigate the effect of endothelin receptors in chronic venous insufficiency (CVI) in lower extremities.

METHODSTen cases of varicose veins from CVI patients (as case group) and ten cases of non-varicose veins (as control group) were investigated in this study. The two groups were divided into two groups respectively: endothelium-intact group and de-endothelium groups. The vasoconstriction mediated by endothelin A (ETA) and endothelin B (ETB) receptors was recorded with myography. The distribution of ETA and ETB receptors was detected by immunohistochemistry method.

RESULTSEndothelin-1 (ET-1) and sarafotoxin 6c (S6c) induced concentration-dependent contraction in the veins. In endothelium-intact veins, the E(max) and pD(2) of contraction curve induced by ET-1 were 132.30% +/- 43.42% and 6.03 +/- 0.35, respectively in control group;and were 19.24% +/- 12.94% and 6.78 +/- 0.46, respectively in case group. The E(max) and pD(2) in case group were much lower than in control group (P < 0.05). The E(max) and pD(2) induced by S6c were 30.10% +/- 12.90% and 6.54 +/- 0.36, respectively in control group, and were 9.61% +/- 1.32% and 6.75 +/- 0.29, respectively in case group; The E(max) in case group was lower than in control group (P < 0.05). In de-endothelium veins, E(max) and pD(2) of S6c were 146.18% +/- 32.33% and 6.50 +/- 0.17 in control group, and 32.93% +/- 3.00% and 6.69 +/- 0.39 in case group; The E(max) in case group was significantly lower than in control group (P < 0.05). ETA receptors was located in endothelium mainly, and ETB receptors in smooth muscle cells mainly. The sites of both ETA and ETB receptors were decreased in case group obviously.

CONCLUSIONSThe contraction mediated by ETA receptor and ETB receptor was decreased with a decrease of ETA receptor and ETB receptor sites in varicose veins of CVI. The contraction insufficiency and down-expression of ETA receptor and ETB receptor are correlated with CVI.

Adult ; Endothelin-1 ; pharmacology ; Humans ; In Vitro Techniques ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Receptors, Endothelin ; drug effects ; physiology ; Vasoconstriction ; drug effects ; physiology ; Vasoconstrictor Agents ; pharmacology ; Venous Insufficiency ; physiopathology ; Viper Venoms ; pharmacology

OBJECTIVETo explore the clinical effect of Agkistrodon antithrombogenase (AAT) in the treatment of rheumatoid arthritis (RA) and its possible mechanism.

METHODSBesides the conventional non-steroid anti-inflammatory agents and disease-modifying anti-rheumatic drug, patients were treated supplementally with intravenous injection of AAT. The intracutaneous test showed allergic to AAT patients were treated with Salvia injection and taken as control group. Changes of related clinical indexes in the two groups were observed.

RESULTSAfter 3 weeks treatment, condition of patients in both groups were improved clinically in joint swollen index, joint tenderness index, morning stiffness time, pain assessment (VAS) and health assessment questionnaire (HAQ) on daily life activity as well as ESR level (P < 0.05 or P < 0.01), with the VAS, HAQ and fibrinogen levels more significantly improved than those of control (P < 0.05 or P < 0.01), and the total effective rate higher in the AAT treated group than those in the control group (P < 0.05).

CONCLUSIONAAT has good effect on easing clinical symptoms of RA possibly through anti-inflammation and improving the microcirculation with less toxic and adverse reaction, so is worthy of recommendation.

Adult ; Ancrod ; therapeutic use ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; Crotalid Venoms ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies

We report the computed tomographic and angiographic findings in the case of a recently obtained successful clinical outcome after embolization of the hepatic artery in the case of a snakebite causing hemoperitoneum associated with hepatic necrosis and rupture with active bleeding.
Aged, 80 and over ; Contrast Media/administration & dosage ; Embolization, Therapeutic/*methods ; Female ; Fibrin Foam/therapeutic use ; Follow-Up Studies ; Hemoglobins ; Hemoperitoneum/*etiology/therapy ; Hemorrhage/etiology/therapy ; Hepatic Artery/radiography ; Humans ; Korea ; Liver/*injuries/pathology/radiography ; Massive Hepatic Necrosis/complications/*etiology/therapy ; Radiographic Image Enhancement/methods ; Rupture, Spontaneous ; Snake Bites/*complications ; Tomography, X-Ray Computed/*methods ; Treatment Outcome ; Viper Venoms/adverse effects

AIMTo determine the involvement of extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in the expression of endothelin receptor type B (ETB) during culture.

METHODSSB386023, a specific inhibitor for ERK1/2 pathway, was used to define the intracellular signaling pathway for the upregulation of ETB receptors and sarafotoxin 6c (S6c), a selective agonist for ETB receptors, induced contraction in isolated rat superior mesenteric arteries. The contraction was recorded by a sensitive in vitro myograph and the receptor mRNA was quantified by a real-time PCR. The phosphorylated ERK1/2 proteins were analyzed by phosphoELISA assay.

RESULTSS6c induced strong contractile responses of the artery after culture for 24 h, while there was no response to S6c in fresh vessel segments. The enhanced contractile response to S6c paralleled with an increase of mRNA for ETB receptors. The phosphorylated ERK1/2 proteins significantly increased after culture for 3 h. After co-culture with SB386023 for 24 h, S6c-induced contractions significantly decreased with reduction of Emax from (217 +/- 14) % to (127 +/- 23) % (P <0.01). This response paralleled with a decreased level of ETB receptor mRNA.

CONCLUSIONERK1/2 pathway was involved in the up-regulation of ETB receptors on smooth muscle cells isolated from rat mesenteric arteries during culture.

Animals ; Cells, Cultured ; Male ; Mesenteric Arteries ; cytology ; Mitogen-Activated Protein Kinase 1 ; antagonists & inhibitors ; metabolism ; Mitogen-Activated Protein Kinase 3 ; antagonists & inhibitors ; metabolism ; Muscle Contraction ; drug effects ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Organ Culture Techniques ; Phosphorylation ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptor, Endothelin B ; biosynthesis ; genetics ; Signal Transduction ; Up-Regulation ; Vasoconstrictor Agents ; pharmacology ; Viper Venoms ; pharmacology

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