AIMTo establish a new and simple flow injection method for the rapid determination of acetylspiramycin (ASPM).

METHODSASPM was determined by chemiluminescence (CL) method combined with flow injection (FI) technology, which was based on the inhibitive effect of ASPM on the chemiluminescence reaction of the luminol-K3Fe (CN)6 system.

RESULTSThe decrease of chemiluminescence intensity was proportional to the logarithm of ASPM concentration (0.1-100) microgram.L-1, the detection limit was 40 ng.L-1 (3 sigma). The whole process, including sampling and washing, could be completed in 0.5 min with a RSD less than 3.0% (n = 5).

CONCLUSIONThe FI-CL method is of both high sensitivity and good selectivity giving a throughput of 120 h-1. The proposed method was applied successfully to the determination of ASPM in pharmaceutical preparations and human urine without any pre-treatment. It was found that the ASPM concentration reached its maximum after being orally administrated for two hours.

Anti-Bacterial Agents ; analysis ; blood ; urine ; Flow Injection Analysis ; Humans ; Luminescent Measurements ; Luminol ; chemistry ; Male ; Microchemistry ; Spiramycin ; analogs & derivatives ; analysis ; blood ; urine

Acute Disease ; Anemia, Refractory ; etiology ; therapy ; Child ; Child, Preschool ; Cord Blood Stem Cell Transplantation ; methods ; Female ; Histocompatibility Testing ; Humans ; Leukemia ; complications ; pathology ; therapy ; Male ; Treatment Outcome

OBJECTIVETo study the stereoselectivity of R-(+) and S-(-)-propranolol glucuronidation and metabolic interaction between R(+)- and S-(-)-propranolol.

METHODSA RP-HPLC analytical method was developed for determination of R-(+)-and S-(-)-propranolol glucuronide (PG) incubated with rat hepatic microsome induced with phenobarbital (PB). The method was applied to investigate the stereoselectivity metabolism of racemic propranolol glucuronidation in vitro.

RESULTIn control and PB group, the concentration of R-(+)-PG produced at different substrates was higher than that of S-(-)-PG. Compared with the control, the V(max) and Cl(int) for R(+)-and S-(-)-propranolol increased significantly the K(m) for R(+)-propranolol was elevated, while that for S-(-) propranolol was decreased.

CONCLUSIONThere is a stereoselectivity in glucuronidation of propranolol in rat hepatic microsome induced with PB and R-(+)-propranolol is preferred. Metabolic interaction between R-(+)-and S-(-)-propranolol exists with a concentration-dependent mode.

Animals ; Enzyme Induction ; Microsomes, Liver ; enzymology ; Phenobarbital ; pharmacology ; Propranolol ; analogs & derivatives ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

Objective To observe the clinical efficacy of electroacupuncture plus oral administration ofZeling Guanjie Xiaozhong Heji in treating rheumatoid arthritis (RA) due to liver-kidney yin deficiency.Method Totally 126 patients with active RA due to liver-kidney yin deficiency were randomized into a treatment group and a control group, 63 cases in each group. The control group was prescribed with orally taking Methotrexate tablets and Leflunomide tablets, while the treatment group was intervened by electroacupuncture plus oral administration ofZeling Guanjie Xiaozhong Heji in addition to the medications given to the control group. The Disease Activity Score 28 (DAS28) was evaluated before and after intervention, and the therapeutic efficacies were compared based on the criteria of the American College of Rheumatoid (ACR) and syndrome of traditional Chinese medicine (TCM).Result The ACR total effective rate was 87.3% in the treatment group versus 65.1% in the control group, and the difference was statistically significant (P<0.01). The total effective rate based on TCM syndrome was 87.3% in the treatment group versus 73.0% in the control group, and the difference was statistically significant (P<0.05). There was a significant difference in comparing the DAS28 score between the two groups after intervention (P<0.01).Conclusion Electroacupuncture plus oral administration of Chinese medication and western medication is an effective approach in treating RA due to liver-kidney yin deficiency, and it can significantly enhance the therapeutic efficacy based on ACR20 and TCM syndrome.

Blood Cell Count ; Bone Marrow Cells ; metabolism ; Child ; Cord Blood Stem Cell Transplantation ; Cytokines ; therapeutic use ; Histocompatibility Testing ; Humans ; Male ; Myelodysplastic Syndromes ; blood ; therapy ; Treatment Outcome

Objective To explore the safety of collection of peripheral blood stem cells(PBSCs) from low-weight infants with osteopetrosis(OP) and its clinical significance. Methods One case of low-weight infants with OP received PBSCs collection using a continuous-flow blood cell separator,and the safety of collection process was observed.The amount of monocyte cell(MNC) and CD34+ cell were noted and its clinical significance was analyzed.Results Low-weight infants with OP could tolerate collection process,the number of collection MNC and CD34+ cells were 10.06?108/kg,2.74?106/kg.Conclusion Adequate PBSCs can be collected from OP who need not be mobilized,thus can offer backup for graft failure.PBSCs collection from low-weight infants is safe.

OBJECTIVETo establish a quantitative analysis method for analyzing the nucleosides in Cordyceps sinensis with capillary electrophoresis, and compare the difference between natural and the cultured C. mycelia.

METHODCapillary zone electrophoresis method was employed to quantitate the adenosine, uridine, guanosine and inosine in C. sinensis, with 0.25 mg x L(-1) boric acid-sodium hydroxide buffer, pH 9.5. The working voltage was 20 kV, the temperature was 25 degrees C, and the detection wavelength was 260 nm.

RESULTWith the capillary zone electrophoresis method, the average recovery of the above 4 nucleosides was 98.9%, 95.1%, 97.8% and 98.8% respectively, with the RSD 0.4%, 1.7%, 1.3% and 5.0%. There was no adenosine in natural C. sinensis and no inosine in the cultured C. mycelia detected.

CONCLUSIONThis method can be used to determine the adenosine, uridine, guanosine and inosine in C. sinensis. The nucleosides in C. sinensis produced from Qinghai province and cultured C. mycelia are obviously different.

Adenosine ; analysis ; Animals ; Cordyceps ; chemistry ; classification ; Culture Techniques ; Electrophoresis, Capillary ; methods ; Guanosine ; analysis ; Inosine ; analysis ; Lepidoptera ; chemistry ; Uridine ; analysis

OBJECTIVETo study the clinical efficacy of transplantation of human neural progenitor cells (hNPCs) in the treatment of severe cerebral palsy (CP) in children.

METHODSForty-five children with CP were voluntarily accepted transplantation of hNPCs. The cells obtained from the forebrain of 10 to 12-week-fetus were cultured and amplified into hNPCs. Then the hNPCs were injected into the cerebral ventricle of the patients with the supersonic guidance.

RESULTSDyssomnia, irritability and muscular tension were improved in one patient 3 days after transplantation. The clinical improvements were observed in the majority of the patients 1 month after transplantation. The therapeutic effects slowed down 3 to 6 months after transplantation. One year after transplantation the gross and fine motor skills and the congnition ability in the transplantation group were considerably surpassed to those in the control group. No delayed severe complications were observed after transplantation. No tumorigenesis was noted 5 years after transplantation.

CONCLUSIONSThe transplantation of hNPCs as a novel therapy is effective and safe for severe CP. Many investigations are needed to evaluate the effect of the therapy.

Cerebral Palsy ; therapy ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Neural Stem Cells ; transplantation

Human Zona Pellucida(ZP), which is a complex matrix surrounding oocytes,is comprised of three immunologically distinct glycoproteins(hZP1, hZP2 and hZP3). Because hZP3 possesses the sperm receptor activity and the acrosome-inducing activity, it has long been used as a candidate antigen to develop an immunocontraceptive vaccine. However, a large amount of native hZP3 protein is unavailable. It is an effective way to express hZP3 protein directly in vitro. Nevertheless, it had been reported that the rhZP3 protein produced in Pichia pastoris was not secreted but accumulated in the cells and could only be purified after being solubilized by strong denaturants. More unfortunately, after purification the final product required 6mol/L urea to maintain solubility. An improved project was advanced with the aim to express secreted and soluble rhZP3 protein in yeast. In this study, the fragment of hZP3 cDNA coding for aa 23 - 408, which the N-terminal leader was removed and most of the C-terminal transmembrane-like domain was reserved, was amplified by two PCR primers including EcoR I and Not I sites respectively and a His6 codon cassette was added to 5'-terminal. The hZP3 insert was incorporated into expression vector pPIC9K. The resulting recombinant yeast expression vector was designated pPIC9K-rhZP3. Linearized pPIC9K-rhZP3 was transformed into Pichia pastoris. After G418 selection, the recombinant Pichia pastoris strains were identified by PCR and the rhZP3 was expressed following the manufacturer' s protocol. Following induction with methanol, the rhZP3 protein was secreted and dissolved into the culture supernatant. SDS-PAGE and Western blot analyses showed that the apparent molecular weight of the expressed rhPZ3 proteins in yeast was smaller and a little size heterogeneity than native ones; after purified with Ni-chelating affinity chromatography, the final product's apparent molecular weight was about 32 - 34KD and their yield more than 20mg/L. We supposed that the C-terminal transmembrane-like domain be useful for secretion of rhZP3 into the culture supernatant and the expressed rhZP3 protein be incompletely digested by proteinases of Pichia into shorter fragments which all were glycosylated inhomogeneously. Fortunately, the fragments of rhZP3 protein can be recognized in Western blot by the polyclonal antibodies to porcine ZP3 which has showed a cross-reactivity with human ZP in vitro. It will be expected that the rhZP3 protein expressed in Pichia pastoris not only has immunogencity, say, it can rise antibodies in vivo to prevent spermatozoa-ovum binding, but also does not contain ovarian factors that might be the cause of undesired side effects, e.g. ovaritis and can be used as a safe immunogen in human antifertility vaccine research.
Blotting, Western ; Chromatography, Affinity ; Egg Proteins ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; genetics ; Humans ; Membrane Glycoproteins ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; metabolism ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism ; Zona Pellucida Glycoproteins

OBJECTIVETo investigate the characteristic and influential factors of the degradation of residual pesticides and alkaloids in Radix Sophorae Flavescentis by H2O2.

METHODThe spiked samples were treated in H2O2 in different reaction time, concentration and pH value. The pesticide residuals were determined by GC-MS, and the contents of alkaloids were determined by HPLC.

RESULTH2O2 had highly activity in degrading organophosphorus and pyrethroid, but had less activity to organochlorines. The degradation processes of organophosphorus and pyrethroid followed first-order kinetics equations, and were influenced by the pH value, the concentration of H2O2 and reaction time. The contents of alkaloids in Radix Sophorae Flavescentis changed not obviously after treatment with 3 mL x L(-1) H2O2 less than 6 hours under neutral condition.

CONCLUSIONH2O2 is a useful reagent for the degradation of organophosphorus and pyrethroid in crude drug.

Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; Gas Chromatography-Mass Spectrometry ; Hydrogen Peroxide ; pharmacology ; Hydrogen-Ion Concentration ; drug effects ; Organothiophosphorus Compounds ; chemistry ; Oxidation-Reduction ; drug effects ; Pesticide Residues ; chemistry ; Plants, Medicinal ; chemistry ; Pyrethrins ; chemistry ; Quinolizines ; analysis ; Sophora ; chemistry