A rapid and sensitive method based on biotin-avidin mediated competitive enzyme-linked immu nosorbent assay(BA-ELISA) was established for the determination of ketamine.The optimal concentration of coated antigen and anti-ketamine monoclonal antibody were found to be 2.0 and 10.2 mg/L.The concentra tions of biotinylated goat anti-mouse IgG(Biotin-IgG) and streptavidin-horseradish peroxidase(SA-HRP) were optimized and the optimum results were found to be 0.29 and 1.0 mg/L, respectively.The linear range of the presented method was from 0.1 to 1000 μg/L, and the limit of detection was found to be 0.03 μg/L.The recoveries of ketamine spiked in human serum and urine were between 94% and 102%.Comparing the result of traditional ELISA, the present BA-ELISA method had a lower detection limit for ketamine.The experimental results indicated that the present BA-ELISA method was specific and sensitive.

We summarized our accumulated clinical and teaching experiences and explored the regularity of acupuncture needling and teaching. It is of great importance in pressing hand during inserting needle. Stroking and pressing are two crucial parts which deserve more attention, and seldom useage of pressing hand should be abolished. Operating hand needs practice before inserting needle, while it should fully relaxed during inserting. Blending "touching", "stretch" "gathering" "erupting" and "advancing" in single moment, applying appropriate dynamic mode of inserting needle such as "join 3 forces as one" "3 points in a line" expertly and naturally. In addition, enough attention should be paid on "altering direction" and "shifting point". Inserting deftly and powerfully, no/slight sensation, deqi when inserting needle are the highest reflection as an acupuncturist.
Acupuncture ; education ; Acupuncture Therapy ; instrumentation ; methods ; China ; Humans ; Teaching

Advance on chemical constituents and pharmacological activities of Stellera plants have been conducted. The chemical constituents include terpenes, coumarins, flavonoids, lignans, volatile oils, and other compounds. Pharmacological studies showed that diterpenoids and biflavones showed strong activities, such as antitumor, anti-HIV, and immune regulations. This review hopes to provide a scientific basis for further research and explorations of the medicinal values of the genus.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Molecular Sequence Data ; Molecular Structure ; Thymelaeaceae ; chemistry ; classification

Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase.Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar lar-vae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR.And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization.Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining.The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR.Results The reverse transcription PCR showed that the ampli-fication products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall.β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of im-munohistochemical staining.The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut.And it was of statistical difference in gene expression level of β-glucosidase among these organs (P <0.05).Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase.Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

Adolescent ; Child ; Cord Blood Stem Cell Transplantation ; Female ; Flow Cytometry ; Humans ; Leukocyte Count ; Male ; Siblings ; Transplantation, Homologous ; immunology ; Treatment Outcome

ObjectiveTo investigate the clinical features and risk factors of patient with Wegener's granulomatosis complicated with pulmonary infection.Methods Patients with Wegener's granulomatosis admitted to our hospital in the past 11 years were retrospectively analyzed.Comparisons between groups were performed by t tests or Fisher test.ResultsPulmonary infection occurred in 27 cases with an incidence rate of 29％.Twenty-six percent of pulmonary infections occurred at the initial diagnosis,and 44％ occurred within 6 months,while 30％ occurred later than 6 months.The clinical manifestations of pulmonary infection were productive cough (89％),hemoptysis (63％),fever and fatigue (56％),chest pain and pactoralgia (33％).The most common causative pathogen were bacteria(59％ ),fungi(37％ ),and tubercle bacillus(37％ ).Sinus infection(P=0.01),hypoproteinemia(P=0.03),hypoimmunoglobulinemia (P=0.007),and methylprednisolone pulse therapy(P=0.002) were the risk factors for pulmonary infection.ConclusionThe occurrence of Wegener's granulomatosis complicated with pulmonary infection is high within 6 months.The most common clinical manifestation is productive cough.The most common causative pathogens are bacteria,tubercle bacillus and fungi.Sinus infection,hypoproteinemia,hypoimmunoglobulinemia,and methylprednisolone pulse therapy are risk factors of pulmonary infection.

The photolytic behavior of deoxyadenosylcobalamin and methylcobalamin in water solution was investigated with high performance liquid chromatography. The results indicated that the photolytic cleavage rate increased with the light intensity, according to which a new method was developed to determine the concentration of vitamin B12 in fermentation broth. The samples were completely photolyzed after cell disruption. The content of vitamin B12 was obtained by determining the content of the hydroxycobalamin. The method shows many advantages, such as rapidness, high accuracy and low sample quantity needed, over traditions methods. The developed method may be used in the field of vitamin B12 fermentation.

ObjectiveTo investigate and compare the behavioral changes, neuron loss of hippocampus and mossy fiber sprouting between pilocarpine-induced status epilepticus (SE) model and pentylenetetrazole (PTZ) kindling model in rats.MethodsAfter two different epilepsy models were made, Vedio was adopted to observe the behavioral changes. Nissl staining and Neo-timms' staining were separately used to observe and compare the neuron loss of hippocampus and mossy fiber sprouting in the dentate gyrus (DG) at different time points during epileptogenisis.ResultsNo recurrent spontaneous seizure, no neuron loss and no mossy fiber sprouting were found in PTZ kindling model; whereas obvious neuron loss was found in CA1, CA3 of hippocampus and hilus of DG, and mossy fiber sprouting were found in pilocarpine model in parallel with recurrent spontaneous seizures. ConclusionPTZ kindling model resembles absence epilepsy in human, while pilocarpine-induced status epilepticus model resembles chronic temporal epilepsy in human. Neuron loss and mossy fiber sprouting may play an important role in epileptogenisis. Pilocarpine-induced epilepsy model can be regarded as an ideal chronic temporal epilepsy model.

Objective　To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolatin g and culturing in vitro. Methods　Common cell culture techn ique, light and electron microscopy were used to study the effects of the growth , proliferation, morphology of the bone marrow mesenchymal stem cells in differe nt adherent time, concentration of serum and cell density. Results　The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×104/ml cell density. The cells were sp indle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cell s in electron microscope. Conclusion　The best condition for iso lation and culture of bone marrow mesemchymal stem cells was successfully establ ished and some biological features were obserred. It found a base for further in vestigation and using of mesenchymal stem cells.