OBJECTIVE:To evaluate the quality of Bifonazole Gel from different manufacturers. METHODS:HPLC was per-formed on the column of Diamonsil C18 with mobile phase of methanol- water- tetrahydrofuran(75∶24∶1,V/V/V)at a flow rate of 1.0 ml/min,detection wavelength was 254 nm,column temperature was 23 ℃,and the injection volume was 10 μl. RESULTS:The linear range of bifonazole was 50-600 μg/ml (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 97.71%-101.68%(RSD=1.16%,n=9). The contents of Bifonazole gel samples from 3 manufacturers were more than 10 mg/g,with little difference. CONCLUSIONS:The method is simple and accurate with good separation and re-producibility,and suitable for the content determination of Bifonazole gel. The investigated contents of products from different man-ufactures conform to relevant standards.

Objective: To isolate small molecular polypeptides which bind to KDR specifically using C7 and 12 peptide libraries. Methods: KDR/IgGFc was coated directly on plates, C7 and 12 peptide libraries are then applied, followed by vigorous washings in washing buffers to remove most non-specifically bound phage and to select specific phage particals by VEGF suspension and elution in acid buffers. The positive phage clones were detected by ELISA, cell-ELISA and competitive binding ELISA. Results: After three washes, 12 positive phage clones were selected and sequenced. Conclusion: A conserved peptide motif was not found in these sequences. The peptide binding to KDR specifically may be helpful for lead drug to improve selectivity and decrease the side effect of anti cancer drug in clinical treatment.

Objective To collect and summarize long-term complications of glucocorticoid (GC) in treatment of Diamond-Blackfan anemia (DBA).Methods A total of 17 DBA patients,treated with GC more than 1 year from December 2009 to November 2012 in Being Children's Hospital,Capital Medical University,were retrospectively investigated.The data of general information,drug treatment,treatment response,height and body mass in different therapy periods,and therapy related adverse reaction were collected.The data entry and the statistical analysis were performed using SPSS 16.0 software.Results Seventeen cases which fulfilled the research criteria were enrolled.The 58.8 percent of cases (10/17) began GC therapy from the age younger than 6 months.The 76.4 percent of the cases (14/17) started prednisone therapy with the dosage ≥2 mg/(kg · d),and the median time of maintenance therapy with this dosage was 2 months (1-5 months).The median time of prednisone dosage greater than 0.5 mg/(kg · d) was 6 months (3-48 months).Patients were divided into 2 groups at the beginning of therapy according to whether their age was younger or older than 6 months.The median height of younger age group was-1.0 SD (-3.5-1.0 SD) of corresponded age-sex-standard height at the beginning of prednisone therapy,and was dropped to-3.5 SD (-3.5--2.0SD)afterhalfyeartreatment.For older age group,it was0.0 SD(-1.5-2.0 SD)and-0.5 SD (-1.5-0.5 SD) respectively.During the therapy,there were 1 fracture,2 measles pneumonia,3 pneumonia,3 hirsuitisms,5 thrushes,and 12 central obesity cases.Conclusions GC related adverse reaction might appear when applying prednisone for the treatment of DBA in long term.It was suggested that GC therapy should start after 6 months old if possible,and the duration of 0.5 mg/(kg · d) GC treatment should be reduced as short as possible.

Objective To analyze the levels and speciation of arsenic metabolites in urine of rats treated with sodium arsenite and sodium arsenate in order to investigate the different aspects of metabolism between sodium arsenite and sodium arsenate,thus to understand further the basic data about relationship between it's metabolism and mechanism of toxicity. Methods Seventy Wistar rats,weighting 80-120 g,were divided into 7 groups of 10 each,such as normal control group,high,middle and low sodium arsenite group and high,middle and low sodium arsenate group. After the animals were fed for one month,the urine was collected by metabolic cage in 12 hours. Applying the high efficiency liquid chromatography and hydride genesis atomic fluorescence spectroscopy (HPLC-HGAFS),the levels and speciation of arsenic metabolites were determined in urine of rats. Meanwhile,the recovery rate of dimethyl arsinic acid(DMA) would be determined to estimate the degree of accuracy of results. Results The levels of iAs~(3+),iAs~(5+) and DMA in middle sodium arsenite group[(121.66±1.26),(10.26±2.68),(200.91±0.56) μg/L]were higher than the high sodium arsenite group[(113.20±0.75),(5.16±1.32),(147.70±μ0.77)μg/L,all P < 0.05]and low sodium arsenite group[(79.35±2.12),(5.13±2.25),(56.35±1.23)μg/L,all P < 0.05]. The levels of iAs~(3+) and DMA in middle sodium arsenate group[(315.81±1.69),(245.12±1.18)μg/L]were higher than the high sodium arsenate group[(85.03±0.56),(110.34±1.04)μg/L,all P< 0.05]and low sodium arsenate group[(22.97±2.67),(15.75±2.15)μg/L,all P < 0.05]. Compared with sodium arsenate group,the levels of iAs~(3+) and DMA in high and low sodium arsenite group were higher(all P < 0.05) ; and the levels of iAs~(3+) and DMA in middle sodium arsenite group were lower(all P < 0.05). Meanwhile,the average urinary recovery rate of DMA of rats in different sodium arsenite group were 94.80%-102.70%,and the average urinary recovery rate of DMA of rats in different sodium arsenate group were 95.33%-108.40%. Conclusion The speciation and levels of arsenic are influenced by the external exposure dose,and some distinction appeared in the metabolism and metabolic path between sodium arsenite and sodium arsenate in urine in vivo.

Objective To investigate the gene expression of methyl-CpG-binding protein 2 (MeCP2) and methyl-CpG-binding domain 2 (mbd2) in patients with systemic lupus erythematosus(SLE) and their significance in the pathogenesis of SLE. Methods Semiquantitative reverse transcriptase-PCR was applied to detect the mRNA expression of MeCP2 and mbd2 in PBMCs obtained from relieved (n=17), active (n=17) SLE patients and healthy controls (n=17). The correlations were further analyzed among these parameters. Results No significant difference was observed in the expression level of MeCP2 mRNA among active SLE patients, relieved SLE patients and healthy controls (P>0.05). The expression of mbd2 in relieved SLE patients was significantly higher than that in health controls (t=12.8, P<0.01), but lower than that in active SLE patients (t=20.0, P<0.01). The expression of mbd2 positively correlated with SLEDAI (r=0.737, P=0.0001) of patients with SLE, and a positive correlation was also observed between the expression of mbd2 and MeCP2 in healthy controls (r=0.550, P=0.0222). Conclusions The expression of MeCP2 and mbd2 may be mutually constrained in normal human, but this relationship seems to be disturbed in patients with SLE.

Objective To explore the clinical application and diagnostic efficiency of two-dimensional shear wave elastography (2D-SWE) in assessing liver fibrosis of patients with viral and non-viral hepatitis. Methood Seventy-three patients with viral hepatitis and sixty with non-viral hepatitis scheduled for liver biopsy in the third Affiliated Hospital, Sun Yat-Sen University from April, 2011 to January, 2013 were enrolled in this study. The Young's modulus in different fibrosis stages, correlation coefficients of liver fibrosis level and area under receiver operating characteristic curve (ROC) were compared between patients with viral and non-viral hepatitis respectively. Results The hepatic Young's modulus of patients with viral and non-viral hepatitis in S0-1, S2-3, S4 were 6.1(4.8-6.6)kPa,7.4(6.0-8.4)kPa,10.3(7.6-14.0)kPa, and 10.7(8.0-13.5)kPa,24.7(17.4-32.1)kPa,26.8(16.5-31.7)kPa, respectively. The difference of Young's modulus between viral and non-viral hepatitis in S0-1 were statistically significant (Z=-3.45, P=0.001), while not in S2-3 and S=4 (Z=-0.40, -0.06, P=0.686, 0.956). Correlation coefficients of liver fibrosis with 2D-SWE in viral and non-viral hepatitis are 0.964,0.817 ( both P=0.000 ) with statistically significant difference (Z=2.42, P=0.015). The area under ROC for S≥2 and S=4 in viral and non-viral hepatitis were 0.964 and 0.930,0.817 and 0.906 respectively. The comparison was significantly different for S≥2 (Z=-2.47, P=0.014), while not for S=4 (Z=-0.502, P=0.616). Conclusion In liver fibrosis assessment, the diagnosis efficiency of 2D-SWE in patients with viral and non-viral hepatitis was different and dependent on fibrosis stage.

OBJECTIVEThis study observed the curative effect of sustained negative pressure drainage application on treatment of severe maxillofacial and neck space infection.

METHODSIncision and drainage were performed to treat 18 patients with severe maxillofacial and neck space infection. A small incision was made on the site of the most obvious swelling or fluctuations, and localized negative pressure was applied with a drainage device on the wound during suturing.

RESULTSAmong the 18 patients, 14 were healed, whereas 4 underwent dehiscence of the wound after the operation. Negative pressure was lost as the drainage tubes were removed, and non-negative pressure drainage method was used instead. During the negative pressure treatment, swelling and pain did not increase after the operation. Other complications, such as asphyxia, septic shock, or mediastinal abscess, did not occur. All the patients were healed and eventually discharged from the hospital.

CONCLUSIONSustained negative pressure drainage, which is a modified version of the traditional method of incision and drainage, is an alternative treatment for severe maxillofacial and neck space infection. Such treatment reduces patient pain and eases doctor exertion. Thus, this method provides a new therapeutic strategy for severe maxillofacial and neck space infection.

Objective To analyze the correlation of interleukin(IL)-12B gene single nucleotide polymorphism(SNP)rs6887695 with clinical phenotypes(including age at onset,family history,clinical types,gender)of psoriasis vulgaris in Chinese Han population.Methods This study recruited 575 patients with psoriasis vulgaris and 1403 healthy controls.DNA samples were obtained from these subjects.PCR with Taqman fluorescent probe(ABI 7900 system)was performed to analyze the genotype of SNP rs6887695 in IL-12B gene.Statistical analysis was carried out by using the software SPSS 14.0,and Chi-square test was conducted to compare the frequency of the SNP rs6887695 genotypes and alleles between the patients and controls as well as between patients with different clinical phenotypes of psoriasis.Results The frequency of GG,GC and CC genotype of the SNP rs6887695 was 42.61％,45.39％ and 12.0％ respectively in the patients,compared to 34.42％,47.83％ and 17.75％ in the healthy controls(x2 =16.31,P ＜ 0.01);the frequency of G and C allele of the SNP rs6887695 was 65.30％ and 34.70％ respectively in the patients,compared to 58.34％ and 41.66％ respectively in the healthy controls(x2 =16.54,P＜0.01).Significant differences were observed in the distribution of genotypes and alleles of the SNP rs6887695 between patients with chronic plaque psoriasis(n =543)and those with acute guttate psoriasis(n =32,x2 =18.11,12.19,both P ＜ 0.01).Increased frequency of G allele and GG genotype of the SNP rs6887695 were noted in the patients with psoriasis vulgaris compared with the healthy controls,and in the patients with plaque psoriasis compared with those with guttate psoriasis.However,there was no statistical difference in the distribution of SNP rs6887695 genotypes or alleles between 540 patients with adult onset psoriasis and 35 patients with child onset psoriasis,between 102 patients with family history and 440 patients without family history,or between 341 male patients and 234 female patients(all P ＞ 0.05).Conclusions The IL-12B SNP rs6887695 may be associated with the susceptibility to psoriasis vulgaris in Chinese Han population,especially with the susceptibility to plaque psoriasis,but seems unassociated with the age at onset,family history or gender of patients.

Objective　To discuss the development and the invasive transfer mechanism.Methods　Cell culture method, p53 protenin immunohistochemistry, PCR-SSCP examination, zymography, PCM and heterogenic inouclation test inside the nude mice were adopted. Results　p53 genetic mutation existed in human melanoma cells. The fluorescent positive rate of 67KD LN-R on the surface of the melanoma cell and the average flurescent intensity are repectively WM451＞WM983A＞WM1341B＞WM35.Early WM35 did not produce MMPs; WM1341B only produced 72KDa(MMP-2),not 92KDa(MMP-9);Both WM983A at progressive stage and distal transfering tumor WM45 produced 72KDa and 92KDa. WM983A and WM451 were also seen to form evident transfering tumor inside the naked mice.Conclusion　p53 genetic mutation, the production of MMPs and the high indication of 67KD LN-R on the celluar surface are the key factors for the human melanoma cell to obtain invasive transferring abililty.

Objective To study the effects of russel viper venom X(RVV X)on blood coagulation protein.Methods We divide diluted protein into control and RVV-X groups,then use chromogenic substract assay to detect the activation effect of RVV-Ⅹ on coagulation factor Ⅶ,Ⅸ,Ⅹ and antithrombin,plasminogen,with or without activator.Results In RVV-Ⅹ group,the coagulation factor Ⅶ, Ⅸ and plasminogen displayed weakly enhanced chromogenesis,all P