BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have a remarkable differentiation potential and superiority as a type of seed cells,but their application is limited in the presence of certain diseases,such as aplastic anemia and myelogenous neoplasm.The present studies have found that seed cells called muscle-derived stem cells (MDSCs) have brought more and more attention,because of their capability of stir-renewal and multi-diffcrentiation like B MSCs.OBJECTIVE: To explore the biological characterization of the muscle-derived stem cells (MDSCs) from rabbits,and analyze the phenotype.DESIGN,TIME AND SETTING: Cell in vitro observation experiment was performed at the Medical Research Center of Second Affiliated Hospital of Sun Yat-sen University from August 2005 to March 2006.MATERIALS: A New Zealand rabbit (1.5 months old,clean grade) was enrolled for the preparation of Muscle-derived stem cells.Growth medium was DMED-LG added with 10% fetal bovine serum and 10% horse serum and fusion medium was DMEM-LG added with 2% fetal bovine serum.METHODS: The muscle mass was removed from the anesthetized rabbit to isolate MDSCs.These cells were dissociated using three enzymes (collagenase XI,dispase and trypsin) respectively.Sediment was resuspended.Then preplate technique was used.The muscle cell extract was plated on a collagen-coated culture flask with growth medium.The flask was called PP1.PPI was kept overnight in a 37 ℃ incubator containing 5% CO2,After that,the suspension was transferred to another collagen-coated culture flask,which was called PP2.PP3,PP4,PP5 and PP6 were constructed later following the same procedures.The cells adhered in PP6 were collected,plated in 6-well plates,and divided into 2 groups.Growth medium was used in one group,in which the cells were kept growing at a degree of confluence beyond 50%,and fusion medium was used in the other one,in which the cells were passaged with a degree up to 30%.MAIN OUTCOME MEASURES: The cells from PP1 to PP6 were collected,and the characterization was identified preliminary by Flow cytomctry,Immunocytochemistry and Western Blotting.The fusion of cells in PP6 was detected at different confluence degrees and concentration of medium.RESULTS: The cells in PP6 showed ＞ 80% desmin+,＞ 70% Bcl-2+,＞ 95% CD45,which indicated that MDSCs were in a high concentration.The expressions of α-SMA in the cells were decreasing with the Preplate technique used and the cells in PP6 almost had no α -SMA expression.When passaged at a high confluence (＞ 50%) or cultured with low concentrations of serum (2% serum),the cells in PP6 had a strong tendency of fusing into myotubes or cell chains and were skeletal myosin+.CONCLUSION: MDSCs,which are capable of multi-differentiation under a high fusion or low serum conditions,express dcsmin and Bcl-2 highly,but extreruelv little CD45 and no α -SMA.

Objective To study the effects of motor imagery therapy combined with electromyographic (EMG) biofeedback on upper limb function in hemiplegic patients.Methods Sixty hemiplegic stroke patients were recruited and divided into a control group (n=20),an electrical stimulation group (n=20) and a combination group (n=20).All groups received basic medication and routine rehabilitation training once daily for 4 weeks.The electrical stimulation group was also treated with EMG biofeedback,and the combination group with motor imagery therapy plus EMG biofeedback.The Fugl-Meyer assessment (FMA),the modified Barthel index (MBI) and EMG parameters were assessed before and after 2 courses of treatment.Results After 8 weeks of treatment all groups had significantly higher FMA scores and MBI scores,and better integrated EMG values,but the effects in the combination group were significantly better than those in the other two groups.Conclusions Motor imagery therapy combined with EMG biofeedback can more effectively promote recovery of upper limb function in hemiplegic stroke patients.

OBJECTIVE: To establish xiaochuan oral oral solution quality control method.METHODS: Three kinds of main components' contents in xiaochuan oral solution were determined by HPLC.RESULTS: The prepared oral solution was brown liquid,with identification and tests all up to the standards specified in Chinese Pharmacopeia(2005 edition).The linear ranges of guaifenesin,ephedrine hydrochloride,and aminophylline(theophylline) were 7～13 ?g?mL-1(r=0.999 9),4.55～8.45 ?g?mL-1(r=0.999 8),and 10.64～19.76 ?g?mL-1(r=0.999 9),respectively,with the average recoveries at 99.35%(RSD=0.97%),100.40%(RSD=1.02%),and 100.57%(RSD=1.08%),respectively.CONCLUSION: The established quality control method could be used for quality control of it.

50%) or cultured with low concentration of serum,these cells tended to fuse to form myotubes,and were skeletal myosin~+.CONCLUSION: Preplate technique can effectively isolate MDSCs,this provides tissue engineering with a new type of seed cells.

Objective To evaluate the clinical application of a novel hepatitis B virus YMDD mutation DNA diagnostic kit (magnetic beads method kit).Methods A total of 324 HBV clinical serum samples was tested with the magnetic beads method kit and another kind of fluorescence diagnostic kit (boiling method).Accuracy, specificity, and sensitivity were compared.Results The consistency of positive detection rate of two kits was 100％ (95％ CI : 98.0％ ～ 100％), negative consistency was 97.12％ (95％ CI : 92.8％ ～99.2％) and the total consistency was 98.76％ (95％ CI : 96.9％ ～99.7％).Four cases of discrepant samples were confirmed by sequencing, and statistical analysis performed by Kappa test (Kappa =0.975) shows good consistency between the two methods.Conclusions The magnetic beads method kit has good consistency compared to the regular boiling method kit, and the polymerase chain reaction (PCR) detection system contains an internal positive control (internal control) to avoid a false negative resuit, which is more suitable for clinical diagnosis.

Objective To prepare the flurbiprofen orally disintegrating tablets and establish the procedures of controlling the quality.Methods Flurbiprofen orally disintegrating tablets was prepared by wet granulation and it content was determined by HPLC.Results The obtained tablets were fine in the shape and smooth in the surface.Their hardness was 2 kg,the disintegrating time was about 30 s.The linear correlation was in a range of 1.0-10.0 ?g/ml,r=0.999 9.The average recovery was 99.32%,RSD=1.09%(n=7).Conclusion Our methods of flurbiprofen orally disintegrating tablets are simple and feasible in preparation and quality control.

Objective To investigate the effects of Shen-fu injection on pulmonary contusion rabbits. Meth-ods Sixteen rabbits were randomly divided into 2 groups: the treatment group (Shen-fu group) and the control group. The animals were induced the pulmonary contusion models. After 60min, the animals in Shen-fu group re-ceived Shen-fu injection received 5 ml/kg, and those in control group 5 ml/kg LRS. The animals were killed six hours later, the right lung tissue wet-to-dry (W/D) ratio and myeloperoxidase (MPO) activity were obtained,the pro-tein expression of nuclear factor-kappa B (NF-κB) and intercellular adhesion molecule-1 (ICAM-1) were also detec-ted. Results The lung tissue W/D, MPO, and the protein expression of NF-κB and ICAM-1 were decreased evi-dently in Shen-fu group (P<0. 01). The morphologic and ultrastructural damages in Shen-fu group were milder than in control group. Conclusion Shen-fu injection is effective on pulmonary contusion rabbits.

Our study investigated the neurotoxicity of quinolinic acid (QA) to spiral ganglion cells (SGCs), observed the protective effects of N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 and magnesium ions on the QA-induced injury to SGCs, and analyzed the role of QA in otitis media with effusion (OME)-induced sensorineural hearing loss (SNHL). After culture in vitro for 72 h, SGCs were exposed to different media and divided into 4 groups: the blank control group, the QA injury group, the MK-801 treatment group, and the MgCl(2) protection group. The apoptosis rate of SGCs was analyzed by Annexin V and PI double staining under the fluorescence microscopy 24 h later. SGCs were cultured in vitro for 72 h and divided into four groups: the low concentration QA group, the high concentration QA group, the MK-801 group, the MgCl(2) group. The transient changes of intracellular calcium concentration were observed by the laser scanning confocal microscopy. Apoptosis rate in QA injury group was higher than that in blank control group and MgCl(2) protection group (both P<0.05), but there was no significant difference between MK-801 treatment group and blank control group (P>0.05). In high concentration QA group, there was an obvious increase of the intracellular calcium concentration in SGCs, which didn't present in low concentration QA group. In MgCl(2) group, the peak values of the intracellular calcium concentration in SGCs were reduced and the duration was shortened, but the intracellular calcium concentration in SGCs had no significant change in MK-801 group. It was concluded that QA could injure SGCs by excessively activating NMDA receptors on the cell membrane, which might be the mechanism by which OME induced SNHL, while Mg(2+) could protect the SCGs from the neurotoxicity of QA.

A Staphylococcus aureus strain, designated zfb, was isolated from a clinical bovine mastitis case of a dairy cow. Staphylococcus aureus zfb can have resistance to methicillin and no lipase contrast by ATCC 25923. The production of the capsule was assessed by the diffuse colonial morphology in serumsoft agar. A mouse infection model was used to determine the LD50 and the invasiveness of SA zfb. The LD50 of SA 25923 to experimental mice was 10-2.5/mL, and the LD50 of SA zfb to experimental mice was 10-4.33/mL. The purpose to detect characteristics of SA zfb makes it an interesting candidate for the preparation and assay of an avirulent mutants against staphylococcal infections and further investigate on pathogenic mechanism.