Objective To report a case of black-dot ringworm caused by Trichophyton tonsurans in a 3-year-old girl. Methods Lesional hair was obtained from the patient and subjected to direct microscopic examination as well as culture. Subsequently, the isolate underwent morphological, biochemical and molecular biology identification. The extracellular enzymatic activity of the isolate was analyzed. Results Microscopy revealed that the hair shaft was filled with fungal spores. Typical colony of the isolate was grayish-white with downy appearance. Slide culture showed centipede-like, lateral, rod-shaped microconidia. Urease test was positive. The amplification of ribosomal DNA (rDNA) ITS domains by PCR produced a 687 bp-sized fragment which had a 100% homology with the sequences of several Trichophyton tonsurans strains in the GenBank database. The extracellular enzymatic activity analysis showed an increase in the activity of alkaline phos-phatase, acid phosphatase, esterase (C4), β-glucosidase, leucine arylamidase, N-acetyl-β-glucosaminidase and a-mannosidase. Conclusions The pathogenic fungus is identified as Trichophyton tonsurans based on morphological and biochemical features as well as sequence of the ITS region of rDNA, and the child was diagnosed with black-dot ringworm.

OBJECTIVE: To establish a HPLC method for simultaneous determination of metronidazole and chloram. phcnicol in comedo cream. METHODS: The method included a Diamonsil C18 column (250mm ? 4.6mm, 5?m) and methanol water(50 : 50,v/v)as the mobile phase and an ultraviolet detecting wavelength of 280nm .RESULTS:The calibralion curve was linear in a concentration range of ( 10 - 250)?g/ml of both metronidazole and chloramphenicol (r = 0.9 998) .The average recoveries of metronidazole and chloramphenicol were 100.53% , 100.53% and RSD were 0.79% , 0.56% respectively .CONCLUSION:The method for the determination of metronidazole and chloramphenicol was rapid and convenient. It can be used for the quality control of metronidazole and chloramphenicol in cream.

Objective To examine the distribution of Malassezia species in follicular contents and perifollicular superficial skin in patients with Malassezia folliculitis and search for its causative agent. Methods A total of 120 patients with Malassezia folliculitis were investigated. Follicular lesions at three different anatomic sites were selected in each patient. Perifolliclar superficial skin specimens were taken by sterile adhesive tape, and the follicular contents of the same follicle were taken by sterile haemostatic forceps. The above specimens were cultured respectively on media containing rapeseed oil. The isolated colonies were identified by their physiological and morphological characteristics. Results Out of 319 isolates obtained from the perifollicular superficial skin, 247 isolates (77.43%) were identified as M. sympodialis, 40 isolates (12.54%) as M. furfur, 27 isolates(8.46%) as M. globosa and 5 isolates(1.57%) as M. obtusa. Out of 314 isolates obtained from follicular contents, 252 isdates(80.25%) were identified as M. globosa, 57 isolates(18.15%) as M. sympodialis, 4 isolates(1.27%) as M. furfur, and 1 isolate(0.32%) as M. obtusa. There was statistical difference in species distribution between the follicular contents and the perifolliclar superficial skin (P

Objectives To compare the difference between multipoint inoculation and routine method for isolation of pathogenic fungi from nail samples of onychomycosis,and to analyze the epidemiology of pathogenic fungi in those patients.Methods The nail clipping samples from each patient were inoculated onto the plates with Sabouraud's agar,Sabouraud's agar without cycloheximide and medium containing rapeseed oil,respectively,by an approach of at least seven inoculating points in each plate (multipoint inoculation),and onto medium slope in tubes with the same media as above mentioned (routine method).In the multipoint inoculation method,plates with more than 3 colonies were taken for further identification of pathogenic fungi based on morphological and biochemical properties.Results Based on the data from 150 samples of onychomycosis,significant differences were found between multipoint inoculation method and routine method (P

Objective To investigate the distribution of Malassezia species in lesional and non-lesion-al sites of patients with pityriasis versicolor(PV),species-variation in different anatomic sites and in lesions with different pigmentation,and the relationship between various Malassezia species and severity and age of PV patients.Methods A total of629skin specimens taken by sterile adhesive tape from the lesions and non-lesional skin were inoculated on media containing rapeseed oil in113patients with PV.Isolated colonies were identified to species based on physiological and morphological characteristics.Results The isolation rates of Malassezia spp.were not significantly different from both lesions and corresponding non-lesional skin.Among non-lesional sites,the isolation rate was significantly higher in forehead and trunk than that in upper and lower extremities.Five species were identified out of565strains obtained from the patients,including M.sympodialis(44.78%),M.furfur(32.94%),M.globosa(11.68%),M.obtusa(5.84%)and M.restricta(4.76%).Two dif-ferent species were isolated simultaneously from27sites.There was no obvious difference in species distribu-tion patterns between lesions and non-lesional sites.M.restricta was isolated from forehead exclusively.Species-variation was closely linked to lesions with different pigmentation and the age of patients,not to the severity of disease.Conclusion There is neither statistical difference of Malassezia isolation rate and species distribution between lesions and non-lesional skin,nor correlation between disease severity and species-varia-tion.The anatomic sites,the diversity of pigmentation pattern and the age of patients seem to be associated with different Malassezia species.

Objective To observe the effect of conventional therapy plus trimetazidine on alcoholic cardiomyopathy (ACM) and plasma brain natriuretic peptide (BNP).Methods Eighty-six cases of ACM patients were divided into control group and observation group by random digits table method with 43 cases each.Control group was treated with conventional therapy,while observation group was added trimetazidine.Course of treatment was 3 months.The left ventricular ejection fraction (LVEF),left ventricular end diastolic diameter (LVEDD),6 min walk distance,plasma BNP before and after treatment and the efficiency in two groups was observed and compared.Results LVEF,LVEDD,6 min walk distance,BNP before treatment in control group were respectively (37.2 ±7.4)％,(57.6 ±7.4) mm,(312.8 ±21.6) m,(846.2 ±63.7)μg/L,and the indicators in observation group were respectively (38.5 ±8.1)％,(57.1 ±6.8) mm,(316.5 ± 23.9) m,(857.6 ± 61.4) μ g/L.All indicators between two groups was no statistically different (P＞0.05).LVEF,LVEDD,6 min walk distance,BNP after treatment in control group were respectively (43.5 ±8.6)％,(54.3 ± 6.4) mm,(511.6 ± 26.7) m,(679.4 ± 51.3) μg/L,and there were significant difference compared with those before treatment (P ＜ 0.05).LVEF,LVEDD,6 min walk distance,BNP after treatment in observation group were respectively (51.6 ± 9.2)％,(51.2 ± 6.3) mm,(579.3 ± 25.1) m,(536.5 ± 50.6)μ g/L,and there were significant difference compared with those before treatment (P ＜ 0.05).There were significant difference in LVEF,LVEDD,6 min walk distance and BNP after treatment between two groups (P ＜ 0.05).The total effective rate in observation group was 90.7％ (39/43),which was significantly higher than that in control group [72.1％ (31/43)] (x2 =4.914,P ＜ 0.05).Conclusion Adding trimetazidine on the basis of conventional therapy can improve the cardiac function and myocardial remodeling of ACM patients and improve clinical efficacy.

A strain of Prototheca species isolated from a case of meningitis was identified by routine morphologic and biochemical methods as well as amplification of the related genes, in which the 28S large-subunit (LSU) region of ribosomal RNA (rRNA) gene and intergenic space (ITS) were amplified with universal fungal primers. The small-subunit (SSU) rRNA gene was amplified with eukaryote-specific primers and Prototheca genus-specific primers. Then, compared the sequences with the ones posted on BLAST (www. ncbi. nlm. nih. gov/BLAST). The organism choice giving the closest match, up to 99%, was considered the most likely correct identification. It was found that this strain of fungus grew well at 25 ℃ or 37 ℃. Smooth,moist colonies with white color were observed on Sabouraud Dextrose Agar (SDA) and Potato Dextrose Agar (PDA). Microscopically, globular or ovoid cells, a number of round, ovoid shaped endospores could be observed. No hypha, ascus or blastic conidia was found upon cultivation on SDA. Based on the morphological characteristics, this isolate could be identified as Prototheca species. The identity with Prototheca wickerhamii was 2.9 % as demonstrated by the API 20C AUX system. Sequence analysis showed that the ITS gene was proved to be a complex structural region which was not suitable for the identification of Prototheca species, but the LSU and SSU rDNA regions showed 94% and 99.9% sequence identities with Prototheca zopfii var. hydrocarbonea (P. zopfii var. hydrocarbonea) respectively, indicating that the SSU rRNA gene sequence might be more reliable on than the LSU rRNA gene sequence for identification of Prototheca species.

Objective To investigate intraspecific and interspecific variation within Malassezia iso-lates from patients with pityriasis versicolor by random amplified polymorphic DNA (RAPD) analysis, to learn the difference between RAPD analysis and physiological and biochemical methods in the typing of Malassezia species, and to explore the relationship between RAPD patterns and Malassezia species. Methods A total of 47 Malassezia isolates were obtained from 34 patients with pityriasis versicolor, and they were classified into 5 species by morphological, physiological and biochemical features, I.e., M. Fin'fur, M. Obtusa, M. Globosa, M. Restricta and M. Sympodialis. Genomic DNA was extracted from the 47 clinical isolates and 10 reference strains (including 7 species) of Malassezia. PCR was performed using 4 random primers including S22, S24, S25 and S33. RAPD patterns were analyzed by NTSYS software and dendrogram was autogenerated. Results Genomic DNA of most strains was successfully amplified with four primers, espe-cially with primers S22 and S24 that resulted in rather stable and clear DNA bands. A total of 82 fragments were amplified from all tested strains. These strains showed both interspecifie and intraspecific variation. Multiple swains were isolated from different body sites of 4 patients and identified into different species by biochemical and morphological typing; those swains from same hosts occupied contiguous positions in the dendrogram and exhibited a high genetic convergence. Conclusion The phenomenon that different strains from a co-host show a high genetic convergence indicates that species specificity and evolution of Malassezia are closely related to its hosts.

Objective To survey the prevalence of chronic obstructive pulmonary disease (COPD)in urban areas of Hunan province and relevant risk factors and provide a basis of the prevention and treatment for COPD. Methods A questionnaire survey was conducted among 4248 residents, aged over 15, by a simple cluster random sampling method in Changsha, Hunan, Wulipai street North Station community. All the respondents filled out an unified epidemiological survey questionnaire. All of the respondents received examination for lung function. Those respondents showed FEV1/FVC ＜70% were further examined by ECG,X ray inspection for differential diagnosis. The data of epidemiological survey was analyzed by multivariate logistic regression method. Results The response rate was 92%. The total prevalence of COPD was 4. 81%.The prevalence of COPD in the males was 6. 6%, and 3. 0% in the females. The prevalence of COPD in the males was significantly higher than that in the females (x2 = 29. 915, P ＜ 0. 01). The prevalence increased with age increasing (P ＜0. 01). The more the education was, the lower the prevalence of COPD was. Risk factors analyzed with non-conditional logistic were as follow. The odd ratio (OR) for COPD in the age was 1.92(P ＜0. 01) and the odd ratio (OR) for COPD in the sex was 1.81 (P ＜0. 01). The weak lighting in house increased the risk with the OR of 4. 25(P ＜0. 01) and pet feeding further increased the risk with the OR of 12.08(P ＜0. 01). The odd ratio (OR) for COPD in the smokers was 1.74(P ＜0. 01) and the prevalence of COPD was related with smoking intensity (branch years of cigarette). Smoking intensity above 500 increased the risk of COPD. The passive smoking increased the risk with the OR of 16. 39(P ＜0. 01). The odd ratio (OR) for COPD in the paternal family history with chronic pulmonary disease was 2. 13(P ＜0. 01) and 2. 11 (P ＜ 0. 01) in the maternal family history. The odd ratio (OR)for COPD in the education degree was 0. 52(P ＜ 0. 01). Conclusions The prevalence of COPD was high in Changsha city, which might be attributed to the risk factors such as house lighting, pet feeding, cooking,aged, male, smoking, passive smoking, and family history. The education degree was the protective factor of COPD. We should intervene the relevant risk factors of COPD so that the prevalence of COPD might be cut down.

Objective: To observe the impact of high thoracic epidural blockade (HTEB) on atrial autonomic nerve remodeling in dogs with atrial ifbrillation (AF) induced by long-term rapid right atrial appendage pacing and to explore the effect of nerve growth factor (NGF) on atrial autonomic nerve remodeling.
Methods: AF model was established by consistent rapid atrial pacing for 6 weeks. 18 experimental dogs were randomly divided into 3 groups: Sham group, the dogs had no pacing while received normal saline injection; Control group, the dogs had pacing and normal saline injection; HTEB group, the dogs had pacing and 0.5% lidocaine injection for HTEB.n=6 in each group. Atrial myocardium collagen volume fraction (CVF) was examined by Masson staining; sprouting of NGF related protein 43 (GAP43) and tyrosine hydroxylase (TH) were assessed by immunohistochemistry; protein expressions of NGF, GAP43 and TH were measured by Western blot analysis.
Results: Compared with Sham group, HTEB group showed decreased CVF and sprouting of GAP43, TH,P<0.05;reduced protein expressions of NGF, GAP43 and TH,P<0.05-0.01. Compared with Sham group, HTEB group presented increased CVF and sprouting of GAP43, TH,P<0.05-0.01; elevated protein expressions of NGF, GAP43 and TH,P<0.05.
Conclusion: Long-term rapid atrial pacing induced AF dog had inhomogeneous sprouting of atrial myocardial nerve which may cause autonomic nerve remodeling; NGF played the important role in such process. HTEB could effectively inhibit NGF up-regulation and suppress the autonomic nerve remodeling in experimental dogs.