Objective To screen differentially expressed proteins between granuloma- and tinea corporis-derived Trichophyton mentagrophytes strains,and to explore the pathogenesis of T.mentagrophytes in deep and superficial infection.MethodsFour T.mentagrophytes isolates from granuloma and 4 isolates from tinea corporis were cultured in agar plates and small steel rings at 27 ℃ and 37 ℃ respectively, followed by morphologic observation.Eight guinea pigs were immunocompromised by glucocorticoids,and superficially and subcutaneously inoculated with the same amount of fungal suspension to develop an animal model of tinea corporis and granuloma,respectively.Two weeks later,the infection of guinea pigs was confirmed by microscopy,fungal culture and histopathology.Proteins were extracted from a highly toxic granuloma-derived T.mentagrophytes strain and a lowly toxic tinea corporis-derived T.mentagrophytes strain,and subjected to twodimensional electrophoresis,mass spectrometry and identification by using the National Center for Biotechnology Information (NCBI) database.ResultsThe granuloma-derived T.mentagrophytes isolates grew better at 37 ℃ than tinea corporis-derived T.mentagrophytes isolates did,while no significant difference was observed in the morphology of colonies between the two kinds of T.mentagrophytes isolates at 27 ℃.Tinea corporis models were successfully established in guinea pigs with the 8 T.mentagrophytes strains,and the granuloma-derived isolates induced a more intense inflammation than tinea corporis-derived isolates.Granuloma model was constructed only with 3 granuloma-derived strains,which was proved by microscopy,fungal culture and histopathology.A total of 463 ± 20 and 398 ± 17 protein spots were detected,with 62 and 21 upregulated proteins,from granuloma-derived and tinea corporis-derived T.mentagrophytes strains,respectively.Bioinformatics analysis revealed the following meaningful proteins from differentially expressed proteins in granuloma-derived T.mentagrophytesstrains, includingenolase, heatshockprotein, serine/threonineprotease, cellularsignal transduction proteins,energy metabolism-related proteins,cytoskeleton proteins and some hypothetical proteins.ConclusionsThe granuloma-derived T.mentagrophytes strains are more heat-resistant with a higher potency to cause infection compared with tinea corporis-derived T.mentagrophytes strains.Differences exist in the expression of proteins between granuloma- and tinea corporis-derived T.mentagrophytes strains.

ObjectiveTo clone the full-length cDNA of Microsporum canis membrane protein PQ-loop repeat protein(PQ-LRP) gene,so as to investigate the roles of PQ-LRP in the pathogenesis of tinea capitis.MethodsA Microsporum canis strain(A518) from a patient with tinea capitis served as the experimental strain.Rapid cDNA end amplification(RACE) was performed to clone the full length cDNA sequence of PQLRP gene.Bioinformatics methods were used to make a preliminary functional analysis of the gene.Results The cDNA of PQ-LRP gene was obtained with a full length of 1522 bp,including the 5' untranslated region (49 bp),coding region(1080 bp) and 3' untranslated region(393 bp).The coding region encoded a protein precursor including 359 amino acid residues.The cloned cDNA of PQ-LRP gene shared an 81％ nucleotide identity with that of Trichophyton tonsurans and a 79％ nucleotide identity with that of Trichophyton rubrum.Conclusions The full-lengthcDNA of Microsporumcanis membraneproteinPQ-LRP gene hasbeen successfully cloned,which will provide an important basis for further researches into the roles of PQ-LRP in Microsporum canis-associated diseases.

Objective To study the significance of AP-PCR i n identification and subtyping of der-matophytes.Methods Using a pair of random primers,OPAA11(5' -ACCCGACCTG -3' ),and OPD18(5' -GAGAGCCAAC -3' )the DNAs of 64isolates of dermatophytes(9species of 3genera),Sporothrix schenckii and Candida albicans were amplified by AP-PCR and analyse d by electrophoresis.Results Distinct DNA band patterns were observed in diffe rent dermatophyte species.Common major DNA bands were observed in Trichophyton rubrum isolated from different areas with s train difference.Conclusion Using OPAA11and OPD18as primers,AP-PCR may be applied in the identification and subtypi ng of dermatophytes.

Objective To develop a new method for the determination of inorganic sulfide in source water by ion chromatography combined with pulsed amperometric detector.Methods The source water samples were injected into ion chromatography system after being filtrated with 0.22 ?m nylon membrane.100 mmol/L of NaOH-250 mmol/L of the NaOAc-0.5% (V/V) ethylenediamine was used as the eluent,special polarizable anions separation column IonPac 3AS7 was selected and sulfide was separated from other anions by isocratic eluent following by three-waveform pulsed amperometric detection.Results The liner range of determination for sulfide was 5-1 000 ?g/L,the equation of linear regression was y=0.325 4x-1.305 3.The detection limit was 0.5 ?g/L,the recovery rates were in the range of 90.8%-96.7%,and the relative standard deviation was 1.32%.Conclusion The method is simple with good selectivity,high sensitivity and good reproducibility and is applicable to the determination of inorganic sulfide in source water.

Objective To investigate difference of NKG2D receptor expression level on the surface of natural killer (NK) cells in the patients with hepatitis B and its clinical significance.Methods This was a four-arm study with different types of subjects,including patients with chronic hepatitis B (CHB,n =22),HBV carriers (HBVC,n=10),patients with acute hepatitis B (AHB,n=18) and healthy donors (HD,n=18).NKG2D protein and mRNA levels on the surface NK cells in the peripheral blood were examined by reverse transcription-polymerase chain reaction assay.The relationship between NKG 2D expression and serum hepatitis B virus (HBV) DNA level was analyzed.The data were compared by analysis of variance and linear regression.Results NKG2D mRNA expression levels in groups of HBVC, HD, AHB and CHB were 0.96±0.17, 1.03±0.12,1.53±0.30 and 1.51 ± 0.35,respectively; the differences among groups were statistically significant (q=7.586,7.485,7.920 and 7.880,respectively; all P＜0.01).NKG2D protein expression levels in groups of AHB,HD,CHB and HBVC were 0.87±0.14,0.89±0.17,0.67±0.09 and 0.59±0.13,respectively; the differences among groups were statistically significant (q=6.92,7.67,7.53and 8.16,respectively; all P＜0.01).The NKG2D mRNA expression levels on NK cells were negatively correlated with serum HBV DNA viral loads in patients with CHB,AHB or HBVC (r=-0.75,-0.66 and-0.69,respectively; all P＜0.01).The NKG2D protein levels on NK cells from patients with AHB and CHB were negatively correlated with serum HBV DNA levels (r=-0.47 and -0.45,respectively; both P＜0.05).Conclusion NKG2D mediated NK cytotoxicity may play a role in viral clearance in hepatitis B.

Objective To investigate the effects, and possible mechanism of action, of vasoactive intestinal peptide (VIP) on acute lung injury in a rodent model of endotoxic shock. Method Endotoxic shock was induced in Sprague-Dawley (SD) rats by intravenous injection of lipopolysacchaiide (LPS) at 10 mg/kg. Three groups (each group with 10 rats), were given injections of either normal saline (Control), LPS 10 mg/kg (LPS group), or LPS 10 mg/kg + VIP 5nmol (VIP). Samples were collected 6 hours after injection. Indices of lung injury including lung wet/dry weight ratio, protein concentration and neutrophil count in bronchoalveolar lavage fluid( BALF) were derived. Assays of TNF-α,IL-1β, IL-10 in serum and BALF were performed using ELISA. Light and electron microscopy were used to detect histopathological changes in lung tissues. Results The lung wet/dry weight ratio, protein concentration and neutrophil count in BALF were significantly raised in the LPS group compared to the Control group (P ＜ 0.05). These indices were significantly lowered in the VIP group compared to the LPS group, though not to the level of the control group. Concentrations of TNF-α, IL-1β, IL-10 in serum and BLAF also increased in the LPS group compared to the control group (P ＜ 0.0S). Levels of TNF-α and IL-1β were significantly lowered in the VTP group compared to the LPS group (P ＜ 0.05), while levels of IL-10 was significantly raised ( P ＜ 0.05). Histopathological changes due to lung injury were not as severe in the VIP group compared to the LPS group. CondusKms VIP plays a protective role during acute lung injury induced by endotoxic shock in rats. Its mechanism of action may be related to down-regulation of proinflammatory cytokines and up-regulation of anti inflammatory cytokines.

Morphea complicated by Hashimoto's thyroiditis is reported in two sisters.Case 1:a 64-year-old female presented with skin rashes on the anterior neck,trunk and bilateral anterior shins for 5 years,itching skin rashes on the perineum for 4 years,and Hashimoto's thyroiditis for 9 years.Physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Dermatological examination showed pink patches on the neck and breast,sclerosis and atrophy of skin over the back,porcelain-white patches on the perineum.Histopathological findings suggested the diagnosis of morphea on the breast and lichen sclerosus et atrophicus on the perineum.Case 2:a 55-year-old female,who was the younger sister of case 1,suffered from gradual sclerosis and atrophy of skin in the left inframammary region and abdominal region for 4 years,as well as Hashimoto's thyroiditis for 3 years.Similarly,physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Hypopigmentation,sclerosis and atrophy of skin were observed in the left inframammary region,abdominal region and central back region.Histopathological examination suggested a diagnosis of morphea.According to the clinical and histopathological manifestations,periodic acid-Schiff staining and thyroid gland function test results,the 2 cases were both diagnosed as morphea complicated by Hashimoto's thyroiditis.

Objective To explore the significance of FSH1 in the pathogenicity of M. canis. Methods Thirty M. cam's strains from tinea capitis lesions and 30 M. canis strains from tinea corporis lesions were cultured, passaged, and induced by medium containing skin tissue of scalp or foreskin from children. Semi-quantitative reverse transcription (RT)-PCR was carried out to detect the expression of FSH1 mRNA in the firstand fifth-generation M. canis strains, as well as M. canis strains induced by the skin tissues. Results The mRNA expression of FSH1 was higher in M. canis strains derived from tinea capitis lesions than in those from tinea corporis lesions (P < 0.01), but there was no significant difference between the first-generation and fifthgeneration M. canis strains (P> 0.05). The skin tissue from scalp and foreskin induced a significant elevation in the mRNA expression of FSH1 in these M. canis strains (F = 2025.713, 1833.139, both P< 0.01), and the inductive effect of the scalp tissue was different from that of the foreskin tissue (P < 0.01). Conclusions The FSH1 mRNA expression is different in M. canis isolated from different body sites. Local skin tissue has an inductive effect on the expression of FSH1 mRNA, and the inductive effect of scalp tissue is more apparent than that of foreskin tissue.

Objective To study the relationship between genotypes and phenotypes,geographical distribution,and the sites of infection of Trichophyton rubrum(T.rubrum).Methods The genotypes were determined by Southern blotting with a probe amplified from the small-subunit rDNA and adjacent internal transcribed spacer(ITS)regions.The phenotypes of T.rubrum were determined by conventional method.Results Twenty genotypes(DNA type A to T)and5phenotypes(villous,furrowed,granular,powdery,and woolly)were recognized among49strains of T.rubrum.Genotype A prevailed in all phenotypes except granu-lar type.Type B represented the most common genotype among the strains of villous type and furrowed type.Type A took the first place in powdery type and woolly type.All of the type A strains were from Dalian.Seven of9type B strains were from Nanjing.Six type C strains were all from Nanjing.The majority strains of21strains isolated from tinea unguium were type C,most of the16strains isolated from tinea cruris and tinea corporis were type A,8strains from tinea pedis were type B,and4strains from tinea capitis were type C.Conclusion There are certain possible relationships between genotypes and phenotypes,geographical distri-bution and sites of infection of T.rubrum.

Objective To develop the system of tumor recognized tool software based on the examination results of multi- tumor protein chip. Methods On the basis of the diagnostic recognized equation, the examination results were comparison by HTA combining VB script and Javascript language program. Results The functions of the software include dates input of tumor protein chip,recognized results, dates interrogation and record printing of 10 kinds of common tumor. Conclusion The software can be diagnosed automatically with the sorts of tumor, which is a scientific, convenient and efficient tool for tumor clinician.