To investigate the immunological activities of the recombinant human phosphatase D2 (rhPLD2) in vitro and in vivo, especially its ability to reduce inflammatory reactions, the cDNA fragment encoding rhPLD2 was cloned into prokaryotic expression vector pET30a by RT-PCR and the recombinant protein rhPLD2 expressed in E.coli was purified from the inclusion bodies, while the anti inflammatory activity of rhPLD2 was determined by the amount of eosinophils in bronchoalveolar fluid(BALF) and blood and the expression of IL-5 and MMP-9 in lung tissues of guinea pig model of chronic asthma. It was found that the rhPLD2 recombinant protein was obtained from human Daudi cells by cloning to E.coli, which contained no membrane-binding site and signal peptide. The cDNA sequence encoded 631 amino acid residues (GenBank Accession Number: AY178289). The purity of the rhPLD2 approached up to 76% with a bioactivity of 50.9745 units/L (0.9212 g/L). In addition, the anti inflammatory effect of rhPLD2 protein could be demonstrated in the guinea pig model of chronic asthma after treatment with rhPLD2 protein, such as down regulation in the expression of the inflammatory cytokine IL-5. It is concluded that the anti-inflammator activity of the recombinant human truncated PLD2 protein produced from the E.coli plasmid can be demonstrated both in vitro and in vivo.

OBJECTIVE To study whether immunosuppressant(IS,cyclophosphamide,CPA) have the function of prevention and treatment of congenitalautoimmune sensorineural hearing loss or not.METHODS The female guinea pigs were immunized by crude conspecific inner ear antigens(CIEAgs),and took orally IS or not at the same time during gestation.Then the pregnant guinea pig's and their offspring's hearing function were measured and inner ear histopathologic changes were observed.Some of offspring borne by the female guinea pigs which immunized with CIEAgs and not treated with IS showed hearing loss and were treated by IS,and their hearing functions were measured and inner ear histopathologic changes were observed with same techniques.RESULTS After immunizing with CIEAgs,some of female guinea pigs which immunized with CIEAgs and not treated with IS showed different degrees of hearing loss(the thresholds of acoustic nerve compound active potential and cochlear microphonic potentials elevated) and immune inflammations in inner ear tissues.All females guinea pigs in experimental group which immunized with CIEAgs and treated with IS at same time,and their offspring have no any hearing loss and inner ear histopathologic changes.After IS therapy,the hearing function improved(mainly at the low-frequency region) in some offspring guinea pigs,which borne by the female guinea pigs which immunized with CIEAgs and not treated with IS.CONCLUSION IS could effectively prevent offspring's congenital sensorineural hearing loss induced by their mother's special antibodies against inner ear tissues antigens.IS showed effective result for treatment of congenitalautoimmune sensorineural hearing loss,but the curative effect was limited.

Objective To clone and express the fused gene fragment coding rhoptry protein ROP2 and major surface protein P30 from Toxoplasma gondii as a preparation for the construction of the complex ROP2,P30 antigen by gene engineering.Methods The gene fragment encoding P30 was amplified by PCR from T.gondii RH strain and subcloned into the recombinant plasmid pUC119/ROP2 already constructed.The recombinant plasmid pUC119/ROP2,P30 was digested by SacⅠ/HindⅢ and inserted into the same site of expression vector pET28b.The recombinant plasmid of pET28b/ROP2,P30 was transformed to E.coli and expressed under the induction of IPTG.Results The gene fragment 700 bp encoding P30 was obtained from the total DNA of T.gondii by PCR.The recombinant plasmid pET28b/ROP2,P30 was successfully constructed,which was highly expressed in E.coli,a fusion protein with molecular weight of 69 000.Conclusion The fusion gene encoding the rhoptry protein ROP2 and the major surface protein P30 of T.gondii has been successfully cloned and expressed to be an expected recombinant fusion protein ROP2,P30 with molecular weight 69 000.

Objective To study the clinical diagnosis and treatment of malignant gastrointestinal stromal tumor. Methods To retrospectively analyze the clinical data of 28 cases with malignant gastrointestinal stromal tumors underweat surgical treatment . Results The malignant gastrointestinal stromal tumor in adults were more than 50 years old,71.4%(20/28) ,and common clinical symptoms were gastrointestinal bleeding,anemia,and pain. The lesion site: 19 cases of gastric bowel, 8 cases of small intestine, 1 case of colon, radical excision in 22 cases, local excision palliative resection in 5 cases, three cases were multi-visceral resection. Conclusion Malignant gastrointestinal stromal tumor could be diagnosed by the means of endoscopic imaging and clear,and preoperative diagnosis was difficult. Surgical resection was the pathology diagnosis and treatment of primary method,if necessary,to ensure multi-visceral resection of the tumor to prevent recurrence of thoroughness, had important significance.

Objective To develop a rapid and reliable polymerase chain reaction (PCR) procedure to detect the pathogenic fungi in clinical specimens. Methods Skin, nail and hair samples were taken from patients suspected of being infected with superficial mycosis. Pathogens were detected by PCR based on the ITS1 primer, and the results were compared with those from microscopic examination and culture. Results One hundred and twelve patients were recruited in this study. For PCR, microscopic examination and culture the sensitivities were 80.7%, 96.5% and 70.2%, the specificities were 100%, 89.1% and 100%, the positive predictive values were 100%, 90.2% and 100%, and the negative predictive values were 83.3%, 96.1% and 76.4%, respectively. The PCR process could be completed within 24 h. Conclusions PCR assay has good specificity and accuracy, while fungal culture takes 2 weeks to get the results. PCR is helpful for making rapid clinical diagnosis, which leads to the appropriate treatment of superficial fungal infection.

Objective To investigate the potential effect of cetyltrimethyl ammonium bromide(CTAB)separated mannan of cell wall from Candida albicans on the production of IL -6and IL -8in h uman peripheral blood mononuclear cells(PBMC)induced by lipopoly saccharide(LPS).Methods PBMCs were pretreated with differen t concentrations of CTAB mannan(1.000mg /mL?0.100mg /mL?0.010mg /mL)for 24h.LPS(50?g /mL)was added and co-incubated for 24h.And a t 48h,the supernatants were collected.At 24h and 48h,only the super-natants of stimulated by CTABmannan were collected.LPS(50?g /mL)was the positive control,unstimula ted culture medium the negative control.The con tents of IL -6and IL -8in the supernatants were determined by ELISA.Re-sults At 24h and 48h,no IL -6and IL -8were detected in 3different concentration-CTAB mannan groups.LPS could induce IL -6(478.507?24.876ng /mL),IL -8(529.655?53.279ng /mL).The contents of IL -6and IL -8of negative control were not detectable.In 1.000mg /mL CTAB mannan +LPS group the contents of IL -6were(85.620?16.058ng /mL,P=0.004),IL -8were(123.940?20.319ng /mL,P=0.011).In 0.100mg /mL CTAB mannan +LPS group,IL -6(210.086?27.874ng /mL,P=0.007),IL -8(206.798?31.878ng /mL,P=0.022).In 0.010mg /mL CTAB mannan +LPS grou p,IL -6(201.387?32.396ng /mL,P=0.014),IL -8(203.133?36.012ng /mL,P=0.015).Conclusion CTAB mannan of cell wall from Candida albicans could downregulate the production o f IL -6and IL -8from human peripheral blood mononuclear cells induced by LPS.

Objective To rapidly detect and identify pathogenic fungi of some deep fungal infections by PCR.Methods The suspensions of22pathogenic fungi(23strains)were amplified by PCR with fungal universal primers ITS86and ITS4which were labeled by FAM.The precise length of amplified fragments was determined by ABI PRISM TM 377Sequencer and Genescan analysis software,then compared with that of am-plicons of corresponding fungal DNA which were previously extracted.Results(1)Amplification of17pathogenic fungi with ITS4,ITS86resulted in a unique fragment length(except for A.nidulans and A.niger,C.albicans and C.stellatoidea,F.pedrosoi and E.dermatitidis).(2)No significant difference of the length of am-plicons was found between the fungal suspension and control organisms,based on the results of Genescan analysis.(3)The whole process took only6h to complete the detection.Conclusion The combination of fun-gal suspension PCR with ITS fungal universal primers and Genescan analysis might provide an accurate,spe-cific,sensitive,and rapid approach to detect and identify22pathogenic fungi causing deep fungal infections,and hold promise to be applied for the diagnosis of deep fungal infection.

Objective To develop a murine model of chromomycosis by using Cladosporium carrioni and explore the pathogenicity of Cladosporium carrio nii in mice. Methods The suspension of Cladosporium carrioni was inoculated to two groups of mice, the immunocompetent mice and the immunosuppressed mice, b y intraperitoneal route using 1 ml inoculum containing 108 conidia/mL. All mice were sacrificed 30 days after inoculation, and then macroscopic examination, his topathology and fungal culture were performed. Results The morbidity in both g roups was 100% according to the dark brown hyphae and sclerotic bodies found in histopathologic examination and fungal culture. Macroscopic examination found th at the adhesion among the internal organs in immunocompetent mice was more sever e than that in immunosuppressed mice. Histopathologic sections showed that necro sis and inflammatory infiltration in immunocompetent mice were more obvious than those in immunosuppressed mice. Conclusions The virulence of Cladosporium car rionii strains is strong enough to construct experimental murine model of chromo mycosis, and animal passage of the strains is unnecessary. This murine model cou ld be used to study the pathogenesis of chromomycosis.

64 ?g/mL, turbinafine 0.125 ?g/mL, ketoconazole 4.0 ?g/mL, and miconazole 8.0 ?g/mL. Conclusion Based on the morphology of colony on SDA and the characteristic structures under the microscope, this is a case of subcutaneous infection caused by Arthrinium phaeospermum.

Objective To investigate the efficacy and safety of butenafine hydrochloride 1% aerosol in the treatment of tinea pedis,tinea cruris or tinea corporis.Methods A randomized,double-blind,multi-center clinical trial was conducted.Efficacy was assessed in terms of mycological cure,total clinical sign and symptom scores,and clinical response,at baseline,mid-term,end of study,and 2 weeks after treatment.Results One hundred and seventeen patients with tinea cruris or tinea corporis were randomly allocated to individual groups treated with either butenafine 1% aerosol (n = 58,male 53,female 5,age 29.45 ? 11.80,course of disease 3.0 ? 5.0 months) or bifonazole 1% aerosol (n = 59,male 49,female 10,age 34.12 ? 12.98,course of disease 3.0 ? 11.0 months).One hundred and nineteen patients with tinea pedis were also allocated to two groups treated with either butenafine (n = 59,male 59,age 22.97 ? 3.97,course of disease 24.0 ? 36.0 months) or bifonazole (n = 60,male 60,age 23.77 ? 4.12,course of disease 36.0 ? 48.0 months).The cure rates and total response rates were 25.86% vs.40.68%,and 86.21% vs.91.53%,in the study group and the control group,respectively,at the end of study,and 58.62% vs.74.58%,and 96.55% vs.96.61% in 2 weeks following-up,for the patients with tinea cruris or tinea corporis.Also,the cure rates and total response rates were 23.73% vs.25.00%,81.36% vs.78.33%,in the study group and the control group,respectively,at the end of study,and 37.29% vs.41.57% and 81.36% vs.90.00% in 2 weeks following-up,for the patients with tinea pedis.Local adverse reactions were recorded in 13 of butenafine group,and 20 of bifonazole group.The differences of above data between two groups were not statistically significant.Conclusion Butenafine hydrochloride 1% aerosol is effective and well tolerated for the treatment of tinea pedis,tinea cruris or tinea corporis.