This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

Bicuspid ; Dental Pulp Cavity ; pathology ; Humans ; Mandible ; Root Canal Therapy

Objective To study and assess clinical application of two Nickel-Titanium(NiTi)rotary instru- ments,namely ProTaper and Hero 642,in preparation of root canals.Methods 125 teeth were divided into three groups and respectively instrumented by stainless K-files,ProTaper or Hero 642 rotary instruments.All teeth were obturated with lateral condensation method.The efficiency of preparation and obturation was analyzed with radio- graphs.Results NiTi rotary instruments were better in keeping the curvature and flow of curved canals than stain- less K files.There was no transportation,apical blockage and ledge in NiTi groups.The operative time was shorter and endodontic flare-up seldom occurred in NiTi groups.Conclusion The NiTi rotary instrumentation technique could be used to prepare curved root canals effectively and quickly.The future use of NiTi engine-driven rotary in- strument appeared to be promising.

Objective To evaluate the CT findings and classification of the Le Fort fracture. Methods Sixty-two cases with Le Fort fractures were studied with thin-slice high-resolution CT scanning and analyzed with three-dimensional(3D)imaging reconstruction.Results Of the 62 patients,10 had Le Fort type Ⅰ fracture,9 had Le Fort type Ⅱ fracture,8 had Le Fort type Ⅲ fracture,and 35 had various combinations of the three types of Le Fort fractures,including 18 Le Fort Ⅰ+Ⅱ fracture,7 Le Fort Ⅰ+ Ⅱ+Ⅲ fracture and 10 Le Fort Ⅱ+Ⅲ fracture.Fifty-five cases had associated multiple fractures in the maxillofacial region.On 2D CT images,Le Fort fracture manifested as multiple and complex fractures. Though 2D image was better than 3D image in accurately defining tiny fractures and fractures of deep structures,the diagnosis of Le Fort fracture could not be correctly made solely on 2D image.3D CT clearly and stereoscopically demonstrated the entire shape and orientation of Le Fort fracture,thus facilitating the correct classification of Le Fort fracture.Conclusion 3D CT image is important in providing information about the space relationship of Le Fort fracture,thus very valuable for the preoperative planning.

Animals ; Decompression ; Decompression Sickness ; physiopathology ; Electrocardiography ; Electroencephalography ; Electromyography ; Electrophysiology ; Male ; Rabbits

This study was aimed to investigate the modulating effects on telomere length and telomerase activity in K562 cells treated by arsenic trioxide, ginseng saponin, beta-elemene alone or in combination with cyclophosphamide (CTX) and to explore the possible mechanism and new therapy for acute leukemia. Human erythroleukemic cell line K562 was co-cultured with the above-mentioned drugs. Cells were collected after 24, 48 and 72 hours for further detection. Telomere length and telomerase activity were detected by Southern-blot and PCR-ELISA respectively. The effects of these drugs were observed at different concentrations and exposure time. The results showed that (1) ginseng saponin, arsenic trioxide, beta-elemene, or CTX could completely inhibit the telomerase activity of K562 cells at proper concentrations and exposure time. The inhibiting effects were enhanced when the three former drugs were used with CTX. Telomerase activity decreased proportionally with the concentrations and length of time. (2) viability of K562 cells was decreased after being co-cultured with arsenic trioxide, ginseng saponin, beta-elemene and CTX. The level of inhibition depends on the concentration and exposure time. (3) telomere length of K562 cells was 5.36 +/- 0.18 kb. After being co-cultured with those drugs for 72 hours, telomere length was 5.90 kb -6.50 kb, significantly longer than that of control (5.18 - 5.35 kb). It is concluded that arsenic trioxide, ginseng saponin, and beta-elemene can inhibit the growth and telomerase activity of K562 cells. The inhibiting effects were enhanced when they were used in combination with CTX. The depression of telomerase activity may be one of the mechanisms of anti-tumor effect. Less dosage and shorter course can be expected when arsenic trioxide, ginseng saponin, and beta-elemene are used in combination with CTX. When telomerase activity was depressed, the telomere length prolonged a little, indicating K562 cell line may extend telomeres by some alternative way other than telomerase activation.
Arsenicals ; pharmacology ; Cyclophosphamide ; pharmacology ; Drug Synergism ; Humans ; K562 Cells ; Oxides ; pharmacology ; Panax ; chemistry ; Saponins ; pharmacology ; Sesquiterpenes ; pharmacology ; Telomerase ; drug effects ; metabolism ; Telomere ; drug effects

0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.

Objective To develop a rapid and efficient single cell PCR method for the non-invasive prenatal hemophilia A genetic diagnosis.Methods Six STR markers (DXS15,DXS9901,G6PD, DXS1073,DXS1108 and F8Civsl3) closely linked to F Ⅷ gene were examined in 118 healthy individuals and 12 hemophilia A families by muhi-plex fluorescent PCR.Three markers with the higher heterozygote rate and diagnostic rate were chosen and combined with amelogenin gene to establish the four-plex nested fluorescent PCR on single lymphocyte level.Results The single lymphocyte amplification rates of Amelogenin,DXS15,FSCivsl3 and DXSI073 were 94.3%,91.4%,100% and 100% in the normal male,and 100% ,97.1% ,97.1% and 97.1% in the normal female's respectively.The female was heterozygous in DXS1073 and DXS15.However,no allele dropout was found in both markers.Neither the false negative nor the false positive amplification was observed.Conclusion The single cell four-plex nested fluorescent PCR method established in this study is convenient and efficient,and will be hopefully employed in the clinical non-invasive prenatal genetic diagnosis for hemophilia A.

Objective To investigate the number of eosinophils in peripheral blood and total serum immunoglobulin E levels from children during infection of mycoplasma pneumoniae,which may elucidate what mycoplasma pneumoniae plays a role in persistent cough and asthma onset.Methods The number of eosinophils in peripheral blood was counted under microscope and total serum immunoglobulin E levels were determined by ELISA in 20 children with mycoplasmal infection ,30 patients with asthma and 25 control subjects.Results The number of eosinophils and total serum immunoglobulin E levels during mycoplasmal infection were significantly higher than in control group,and lower than in asthma.Conclusion The high eosinophil counts and total serum immunoglobulin E induced by mycoplasma pnemoniae play a key role in persistent cough and asthma attack.

Objective:To invetigate the value of a novel puncture and drainage of intracranial hematoma for the treatment of hypertensive intracerebral hemorrhage.Methods:In the group there were 27 patients with hypertensive intracerebral hemorrhage,with mean age of 61 years. Their hematomas located in thalamus(1 patient),basal ganglia(22 patients),and lobe(4 patients).The mean(SD)hematoma volume was 40(3.2)mL,the mean Glasgow Coma Scale (GCS)score was 10.15,and the mean National Institutes of Health Stroke Scale(NIHSS)score was 30.65 at admission.CT scan provided hematoma location.and the percutaneous puncture. grind and drainage were performed under the local anesthesia by using a novel puncture and drainage of intracranial hematoma.Results:No adverse events occurred during the punctures and after the procedures.One patiems died 20 days after procedure.Other patients were followed up for more than 6 months.Eight patients had a good outcome as assessed by Glasgow Outcome Scale(GOS)scores,15 had mild disability,2 had serious disability,and 1 was in a permanent vegetative state.Conclusions:This novel puncture and drainage of intracranial hematoma can be used in the treatment of hypertensive intracerebral hemorrhage,and it is simple and safe.