Objective To evaluate the performance of microfluidic chips in the identification and genotyping of Malassezia species.Methods This study included 6 reference Malassezia strains and clinical Malassezia isolates from the scrapings of patients with pityriasis versicolor and follicular contents of patients with Malassezia folliculitis.These isolates were identified by DNA sequencing,random amplified polymorphic DNA (RAPD)-PCR and microfluidic chips.Cluster analysis was carried out and tree diagrams were generated.Results A total of 83 Malassezia isolates were obtained from 72 patients with pityriasis versicolor and 11 patients with Malassezia folliculitis.Genomic DNA of most strains was successfully amplified by PCR with two primers S22 and S24,and PCR with S22 primer produced more stable and clear amplification bands than that with S24.Positive bands of different sizes were repetitively obtained by using microfluidic chips,with interspecies and intraspecies polymorphisms observed in all the strains.On the basis of DNA sequencing,microfluidic chips and RAPD-PCR could be used to successfully distinguish the following eight species,i.e.,M.furfur,M.sympodialis,M.globosa,M.pacbydermatis,M.slooffiae,M.Japonica,M.yamatoensis and M.dermatis.Conclusions As a rapid,high-throughput and high-sensitivity method,microfluidic chips combined with RAPD-PCR shows an advantage for analyzing interspecies genetic diversity,genetic relationship of Malassezia species,as well as for identifying new Malassezia species.

A 13 year old male farmer suffered from cutaneous and nail phaeohyphomycosis for five years is reported in this paper. The lesions were dull red, nodules and well defined plaques with ulcerations, mainly on the face, extremities, palmar and plantar surfaces and buttocks. Some of them were covered with thick greyish black crusts. General examination did not reveal abnormal findings except the skin lesions. Histopathology showed granulomatous response with numerous light brown septate branching hyphae. The colonies were greyish brown on SDA and PDA at 25℃ and 37℃ with brownish black ascomata. The ioslated strain was identified as Chaetomium globosum based on the morphological features of ascomata, ascomal hairs and ascospores.

Several years after trauma,a patien t suffered from skin verrucous hyperplasia for more than 30years,accompanied by ulcer occasio nally,on the feet,ankles and legs su ccessively.Mycological examinati on(9times)had been done before treatment.Slig ht septate hyphae and /or spores were demonstrated by direct mi-croscopy(5times);and yellowish green colony grew on S abouraud agar at 25℃for 2times,it wa s identified as Epidermophyton floccosum.Yeast -like colony grew on Sabourau d agar at 25℃for 5times,it was identi fied as Candida ciferrii by API.After treatment,Candida ciferrii could still be detected by mycologic al examination(6times),but no Epidermophyton floccosum was found.Slight septate hyphae were demonstrated in stratum spinosum,stratum granulosum and stratum corneum by histopathological examination.Isolated or clustered blastospores,spores and chlamydospores were demonstrated in stratum c orneum by histopathological examin ation.Therefore,it was concluded that the skin verrucous hyperplasia in this c ase is caused by mixed infection of Epidermophyton floccosum and Candida ciferrii.The patient was treated with flucon azole capsule 50mg daily for5weeks,followed by terbinafine tab let 250mg daily for 36weeks,and then250mg twice daily for 18weeks.In the 5th week of terbinafine therapy,a part of the lesions on the shin of left leg healed.No further therapeutic effect was observed.

Objective To study the distribution of genotypes of Candida albicans isolated from different body sites of patients with candidal vulvovaginitis(CVV).Methods PCR was designed to amplify group I intron-containing region in25S rDNA of Candida albicans.The strains of Candida albicans could be classified into three genotypes:genotype A(~450bp),B(~840bp)and C(~450bp and~840bp),on the basis of different ranges of bands of amplicons.Results Sixty women with CVV were recruited,of whom54were caused by Candida albicans.Among the54patients39had non-recurrent CVV and15had recurrent CVV(RCVV).Candida albicans could be isolated simultaneously from different body sites in32of54patients,including19(19/39)with non-RCVV and13(13/15)with RCVV.A total of92strains of Candida albicans were isolated from vagina,tongue and anus in54patients with CVV.Eighty strains of genotype A,8of genotype B and4of genotype C were found.The same genotypes of Candida albicans in different body sites were identified in24patients,and the different genotypes were identified in8patients.Conclusion Genotype A is predominant in CVV.The other two genotypes(B and C)are not commonly seen,and mainly isolated from non-vaginal sites.The colonization of Candida albicans in the non-vagina sites is more frequent in RCVV than that in CVV,and the intestinal reservoir theory may play a role in the relapse of RCVV.

Neuron is the basic unit of the biological neural system. The Hodgkin-Huxley (HH) model is one of the most realistic neuron models on the electrophysiological characteristic description of neuron. Hardware implementation of neuron could provide new research ideas to clinical treatment of spinal cord injury, bionics and artificial intelligence. Based on the HH model neuron and the DSP Builder technology, in the present study, a single HH model neuron hardware implementation was completed in Field Programmable Gate Array (FPGA). The neuron implemented in FPGA was stimulated by different types of current, the action potential response characteristics were analyzed, and the correlation coefficient between numerical simulation result and hardware implementation result were calculated. The results showed that neuronal action potential response of FPGA was highly consistent with numerical simulation result. This work lays the foundation for hardware implementation of neural network.
Action Potentials ; Computer Simulation ; Humans ; Models, Neurological ; Neural Networks (Computer) ; Neurons ; cytology ; Synaptic Transmission

Objective To investigate the mycologic features and variation in gene sequence of a hot-resistant Trichophyton rubrum which caused granulomatous skin lesion.Methods The isolate was identified by routine and polymerase chain reaction(PCR)technique.Ribosomal conserved sequence and trs-1(tandem repetitive subelement)sequence in the ribosomal non-transcribed spacer(NTS)were determined.Results The hot-resistant isolate was identified as T.rubrum by routine and PCR technique.The trs-1sequence variation in rDNA NTS was found in the strain.Conclusion The trs-1sequence variation suggests that this strain be one of isotypic variants of T.rubrum,which has much higher pathogenicity to cause granulomatous skin lesion,or sequence variation of the strain may be the result of adapting itself to environmental change(37℃),which needs further studies.

Objective To report a case of subcutaneous phaeohyphomycosis caused by Curvularia clavata in China. Methods The skin specimen was collected, and examined by KOH preparation, fungus culture and histopathology. Results A 17-year-old male farmer had dull red, warty and well-defined plaques with some ulceration on his face and his left upper arm for nine years and a history of trauma of his face before the appearance of the lesions. General examination did not reveal abnormal findings except the skin lesions. KOH direct examination showed septate hyphae with irregular branches. On the culture medium the colonies were black in color. Under microscope the conidia were borne singly or in groups at the tip or at the sides of the conidiophores, they were light brown to brown, smooth, straight or slightly curved and clavate, measuring 15 ~ 30 ?m long and 6 ~ 12 ?m thick, with 3 ~ 4 septates. Histopathology showed a granulomatous response. PAS staining showed irregularly swollen or toruloid conidia with branched hyphae. The isolated strain was identified as Curvularia clavata Jain. Conclusion It is the first case report of subcutaneous phaeohyphomycosis caused by Curvularia clavata in China.

This study was aimed to optimize the extraction process of ginseng in Shen-Qi Bu-Qi (SQBQ) granules. The orthogonal experiment method was used to optimize the extraction process of ginseng adopting ethanol concentration, ethanol volume, extraction time and extraction times as factors. The content of ginsenoside Rg1, Re, Rb1 content in the extract was determined by HPLC. And the optimum extraction conditions were determined with indexes of three kinds of saponin yield. The results showed that the optimum extraction process of ginseng for SQBZ granules was that adding 6 times amount of 60% ethanol technology for medicinal materials, extract for 2 times, and 3 h for each time. It was concluded that the optimization of ginseng extraction process was stable, reasonable and feasible with high extraction rate.

Objective To investigate the biological effect of anti-leukemic cells induced by eosinophilic granulocyte (EOS) in bone marrow of patients with chronic myelogenous leukemia (CML).Methods The BCR-ABL fusion gene as well as the expression of IL-12 and IL-17 mRNA were performed by RT-PCR.The serum concentrations of cytokine IL-12 and IL-17 were determined by enzyme-linked immuno sorbent assay (ELISA).Immunochemistry staining and cytochemistry staining were used to observe the peroxydase (POX) and human Leukocyte antigen (HLA)-DR expression of EOS in bone marrow.Immunofluorescence staining was used to observe mannose receptor (MR),IL-12,IL-17A and IL-17receptor A (IL-17RA) expression of EOS.The results between the CML patients and the healthy controls were compared.Results Serum levels of IL-12 and IL-17 were higher in the 60 CML patients [(196.33 ±21.79) ng/L and (36.55-±3.01) ng/L] than those in the controls [(96.60 ±4.92) ng/L and (23.74 ±1.36) ng/L].In the 32 patients with activated EOS,the levels of IL-12 and IL-17 were (273.12 ± 17.16)ng/L and (40.11 ± 6.13) ng/L,which were significantly higher than those in the non-activated EOS [(126.16 ± 14.27) ng/L and (28.14 ±5.29) ng/L] (P values ＜0.01).IL-12 and IL-17 mRNA were expressed in activated EOS,while BCR-ABL fusion gene was not found.The amounts of EOS were increased abnormally in the bone marrow and peripheral blood of the CML patients with POX positive staining in the cytoplasm and weakly positive HLA-DR staining.It was observed easily by a microscope that EOS could attack leukemic cells in bone marrow through adhesion,capture and phagocytosis.Activated EOS could express IL-12,IL-17A and MR,which was related with the serum levels of these cytokines.Conclusions Activated EOS in bone marrow of CML patients could express IL-12 and IL-17.Activated EOS could induce coup injury to leukemic cell by releasing POX and expressing IL-12 and IL-17.It can also capture or swallow target cells via the expression of MR on the membrane.EOS may play an important role in the anti-tumor immunologic function in bone marrow of CML patients.

Objective To develop a microtitration plate en zyme immunoassay to differentiate Candida albicans from Candida dubliniensis isolates using two specific probes s imultaneously.Methods The fun-gus-specific universal primers derived from the internal transcribed s pacer region of fungal rDNA were labeled with biotin,while the C.albicans or C.dubliniensis specific capture probes were coated on the microplates.Genomic DNA purified from the two species was amplified by PCR.The biotinylated p roducts were captured by the probes coated on the microplates.The A 405 value was finally determined by the c olorimetric assay.Results The two species of Candida could be detected specifically.Out of 108clinical isolates originally identified as C.albicans on the basis of germtube formation,two isolates were positive for C.dubliniensis and negative for C.albicans.The other106isolates were positive for C.albicans and negative for C.dubliniensis.Conclusions Two-specific-probe hybridization method is rapid and re liable for differentiating C.albicans from C.dubliniensis.