Objective To study the regulation of quorum sensing molecule tyrosol and farnesol on biofilm formation of Candida albicans. Methods Candida albicans biofilms of clinic isolates and standard strain SC5314 were built when quorum sensing molecule existed. And inverted microscope was used to observe the morphology of C. albicans cells. RT-PCR and MTT assay were carried out to investigate the effect of quorum sensing molecule on expression of the two genes (HTA1 and EFG1) and cytoactive. Results Tyrosol could not promote hyphae development and cytoactive of C. albicans biofilms. The expression of HTA1 of C. albicans in biofilms was up-regulated by tyrosol but EFG1 was not. The inhibitory effect of farnesol on hyphae development, cytoactive and gene expression were not changed by addition of tyrosol. Conclusion Tyrosol can make C. albicans biofilms active in early stage. But when tyrosol and farnesol were simultaneously added, the effect of tyrosol were masked by farnesol. And C. albicans cells were more sensitive to farnesol than to tyrosol.

Objective To analyze plasma vitamin E and CoQ10 levels in patients with autosomal recessive cerebellar ataxia for finding the evidence of the related pathogenesis research and therapeutic strategies.Methods The plasma vitamin E and CoQ10 levels were detected by high performance liquid chromatography (HPLC) with diode array detector in 123 probands of autosomal recessive cerebellar ataxia pedigrees.Quantitation was performed using vitamin E and CoQ10 external standard and two 5-point calibration curve;clinical manifestations were analyzed simuhaneously.Results Vitamin E and CoQ10 levels of healthy subjects in the plasma were (8.77 ± 2.28) μg/ml and (1.31 ± 0.38) μg/ml,respectively;the plasma vitamin E and CoQ10 levels of patients were (5.61 ± 2.04) μg/ml and (0.79 ± 0.26) μg/ml,respectively,which were significantly lower than those in healthy controls (t =11.87,13.15;all P＜ 0.01).Clinical manifestations were characterized by cerebellar symptoms,and gait instability was usually the first recognized abnormality.Most of early onset occurred before the age of 25 years (111/123);dysarthria and abnormal eye movement were observed,with cerebellar atrophy on MRI;concomitant symptoms were also present.Conclusions HPLC analysis shows that the plasma vitamin E and CoQ10 levels of patients with autosomal recessive cerebellar ataxia are generally lower than those in the healthy controls.Several patients with significant reductions in these two levels have genetic defects.The combination of clinical phenotypes,biochemical indexes and genetic analyses will be helpful for the establishment of diagnosis and specific treatment.

OBJECTIVE:To study the quality control,stability and therapeutic effect of compound fleroxacin ear-drops.ME_THODS:The content of fleroxacin in the ear-drops was determined by ultraviolet spectrophotometry.RESULTS:There was a good linearity in a range of concentrations from 2?g/ml to 12?g/ml,r=0.9 998,average rate of recovery was 100.38%,RSD=0.39%(n=6).The preparation showed stable quality and satisfactory effect.CONCLUSION:This ear-drops is simple in preparation,reliable in quality-control and effective in clinical use.

Objective To explore the effect of ankle strategy stability limit training on balance and gait in recovering stroke patients with hemiplegia. Methods Forty recovering stroke patients were randomized into an intervention group and a control group.The patients in the intervention group were given ankle strategy stability limit training using visual feedback on the static long sets of a Smart Equitest Balance Master (SEBM) machine.Those in the control group practiced routine postural balance training using mirror visual feedback in parallel bars.Both groups of patients practiced balance and posture control for 30 minutes,once daily,6 days a week for two weeks. Both groups were also given routine therapy and other rehabilitation.The patients' balance function was evaluated using the Berg Balance Scale (BBS),and their gait was assessed using the walk across technique (WA). Results There was no significant difference between the two groups with regard to general information,BBS scores or WA results before treatment.After 2 weeks of treatment,BBS scores as well as the step length and pace in the WA improved significantly in both groups,but all improved significantly more in the intervention group.There was no significant difference in width of gait. Conclusion Ankle strategy stability limit training can enhance weight-bearing on stroke patients' affected foot as well as their balance and the symmetry of their steps.

Objective To study the role of cell density in the tyrosol production and morphology for Candida albicans biofilms. Methods C. albicans SC5314 and clinical isolates were propagated in yeast peptone dextrose (YPD) medium. Cells were collected by centrifugation and washed twice in sterile phosphate-buffered saline (PBS) before this study, then resuspended in RPMI 1640 supplemented with L-glutamine and adjusted to a desired concentration of 5 × 10~6 cells/ml, 5×10~5 cells/ml, 5 × 10~4 cells/ml, 5 × 10~3 cells/ml after counting with a hematocytometer. Standardized C. albicans cells were prepared as above description and 2000 μl of this standardized cell suspension was dispensed into the wells, then C. albicans biofilms were formed on the bottom of the polystyrene wells. In this study, tyrosol synthesized by SC5314 and clinical isolates of C. albicans biofilm was quantified by high performance liquid chromatography (HPLC). The effects of tyrosol on morphology of C. albicans biofilms were investigated by scanning electron microscopy (SEM). Results Tyrosol production of C. albicans biofilms was affected by cell densities. At lower inoculation size(5 μ 10~3 cells/ml), there was too less tyrosol production to be detected at the early stage of the biofilms formation. At higher inoculation size (5 μ10~6 cells/ml), tyrosol can be detectable at the early stage or at the mature stage of biofilms formation. There was a sharp increase in tyrosol concentration at 24 h, while there was a decrease in tyrosol concentration after that time from the strains when the strains were at an inoculation size of 5 × 10~6 cells/ml and 5 × 10~5 cells/ml. Cell densities affected the morphology formation of the C. albicans biofilms. At the early stage of the biofilms formation, C. albicans grew less germ tube at lower cell densities than that at the higher cell densities. With the mature of the biofilms, C. albicarts grew more hyphae at higher cell densities than that at the lower cell densities. All these above showed that cell densities played an important role in the propagation for the C. albi-cans in the biofilm formation. Conclusion Cell density play an important role in the formation of the C. albi-cans biofilms and the production of the tyrosol.

Objective To express protein transduction domain (PTD)-deletion proline domain (ΔPRD) Foxp3 fusion protein, and to analyze its influence on mixed lymphocyte reaction in mice.Methods We cloned mouse ΔPRD of Foxp3 gene by PCR, and inserted it into pET28a-PTD, pET28a-PTD-eGFP vector, then expressed fusion proteins in E.coli Rosetta (DE3). The fusion proteins were purified and refolded by Profinity IMAC Ni~(2+)-Charged Resin. The expression of fusion proteins was identified by Western blot. Flow cytometry assay was used to detect the effect of PTD-ΔPRD fusion protein to transduce into mouse EL-4 cells. The ability of fusion protein to inhibit the proliferation of EL-4 cells was analyzed by two-way mixed lymphocyte reaction.Results The PTD-ΔPRD fusion proteins were expressed and purified efficiently. Western blot and flow cytometry indicated that PTD-ΔPRD fusion protein was transduced into EL-4 efficiently. Mixed lympocyte reaction assay showed that PTD-ΔPRD fusion protein had the bioactivity to inhibit the proliferation of EL-4 cells.Conclusion The PTD-ΔPRD fusion protein was expressed in E.coli system and could be transduced into cells effectively, suggesting that PTD-ΔPRD fusion protein may be an inhibitor in lymphocytes from mouse spleen.

Objective To investigate methods for prenatal diagnosis of spinocerebellar ataxia type 3 (SCA3).Methods Cordocentesis were performed in the pregnant SCA3 female proband of pedigree during the 20th gestational week.Polymerase chain reaction (PCR) and short tandem repeat (STR) analysis based on CEQS000 sequencer were applied to analyze the CAG repeat of SCA3 gene.Results The proband had 31/75 CAG repeat alleles of SCA3 gene; her spouse had 14/27 CAG repeat alleles; the fetuse had 14/31 CAG repeat alleles (14 repeat from the father,and 31 repeat from the mother),which is in conformity with Medelian inheritance.The fetuse inherited the normal CAG repeat allele from the mother.The above results of the fetuse were verified after its birth.Conclusion Detecting CAG repeat dynamic mutation of SCA3 gene based on umbilical cord blood and STR analysis could be a rapid and reliable method for prenatal diagnosis of SCA3.

Objective:To introduce recent progress on diagnosis and treatment of benign symmetric lipomatosis(BSL).Method:Detailed clinical data of 6 patients with BSL were reviewed and analyzed.We present a summary of the clinical symptoms,physical sign,diagnosis and therapeutic methods of BSL. And related literatures were discussed together.Result:All of 6 patients have excessive subcutaneous fat deposit predominantly around neck.One patients had upper extremity localizations. Six patients had the complication of left ventricular diastolic function changes,glucose intolerance or diabetes mellitus, chronic hepatopathy, hyperuricemia and sleep apnea syndrome in one or more. One patients with several symptoms occur simultaneously, another one female patient was accompanied by all symptoms but chronic hepatopathy. Five male patient were alcohol abusers. Tatal neck lipentomy and abstinence from alcohol were performed on 5 patients. One patient refused treatment.During a follow-up of 3 months to 4 years, one patients was relapsed again,and no recurrence was seen in another 4 patients. All pathological results were nonencapsulated fat. Conclusion:BSL is a lipodystrophy caused by diffuse fatty tissue, symmetry deposition in the neck and shoulder subcutaneous fascia space or deep fascial space .The highest incidence favors to middle-aged man who is alcoholist. Chronic alcohol addiction and typical clinical symptoms help to diagnosis BSL. Lipectomy represents a successful procedure in treating BSL.

Objective To study the effect of Xi Yanping injection on electrocardiogram in Beagle dogs.Methods 24 Beagle dogs were divided into negative control group (0 mg/kg), low-dose group (15 mg/kg), middle-dose group (75 mg/kg), high-dose group (250 mg/kg).Each group consist of 3 animals/sex/group, all animals were administrated by intravenous drip at 20 mL/kg once a day.Administration period was 28-days with 14-days recovery phase.Animals LeadⅡelectrocardiogram ( ECG) was measured during quanrantine, administration and recovery phase.The following parameters included heart rate, P -Rinterval, Q -T interval, P, Q, RS wave amplitude and ST and T wave amplitude were determined, serum biochemical parameters were measured to evaluate the effect on main organs.Results All parameters of quarantine period were fell in the normal limit.After the first administration, heart rate decrease was found in low dose group and P-Rinterval prolongation was found in high dose group.No abnormal was found in dose metaphase,end of dose stage and recovery stage.Conclusion Xi Yanping injection had no obvious influence on ECG of Beagle dog.

Objective To explore the functional activation cerebral areas evoked by mechanical stimulation induced pruritus perception imitating physiological process.Methods Scratching the planta gently to induce pruritus with brush pen wool in a group of 16 healthy volunteers,scanning BOLD-fMRI serials with a block design on a 3.0 T MR machine,and identifying the functional cerebral areas evoked by pruritus with SPM8 software for analysis.Student's t test was used to compare the difference of incidences with P＜0.05 for statistical significance.Results Several brain regions were activated by pruritus stimulus.The strongest activation evoked by pruritus was found at the contralateral thalamus and paracentral lobule(t=5.26,5.23),the most activation volumes were found at the contralateral paracentral lobule,postcentral gyrus and prefrontal lobule(146,151 and 326 volumes).Different degrees of activation were discovered at bilateral insula,preeentral and postcentral gyrus,thalamus,lentiform nucleus and cingulated gyrus.Conclusion The postcentral gyrus,paracentral lobule,insula,precentral gyrus and frontal lobe were the functional activation cerebral areas of pruritus perception evoked by mechanical stimulation.