Peripheral T cells are closely related to nature killer( NK)cells,and share some immunophenotypic and functional properties with NK cells.Peripheral T-cell and NK cell lymphomas are heterogeneous groups of lymphomas,and the subgroup classification is complicated.Currently,the pathogenetic molecular lesions remain not to be deciphered for most entities.However,novel insights into the features of angioimmunoblastic T-cell lymphoma displaying correlation with normal helper T cells,the gene of anaplastic large cell lymphoma,and potential therapeutic biomarkers,have been gained from molecular technology application.

Objective: To evaluate color velocity imaging-quantitative method (CVI-Q) in estimating cerebral hemodynamic change in patients with cerebral infarction. Methods: The carotids of 60 normal people and 40 cerebral infarction patients were detected by CVI-Q. We observed endangium thickness and atheromatous plaques,and measured the diameter (d), peak velocity(Vmax), resistance index(RI) and blood flow volume(Q) of the common carotid arteries. Results: In cerebral infarction group there were 75% cases with endangium thickening to different degrees, 45% cases with atheromatous plaques and 71% plaques in carotid enlargement section or bifurcation. The data measured in 2 groups were compared: (1)The d value in cerebral infarction cases increased than that in normal（P<0.05 or P<0.01）; (2)The Vmax reduced in cerebral infarction cases(P<0.05); (3)The RI increased in cerebral infarction cases (P<0.05 or P< 0.01); (4) The Q value reduced in cerebral infarction cases (P<0.01). Conclusion: CVI-Q can be used for detecting cerebral hemodynamic changes and provide quantitative indexes for clinicians to estimate ischemia degree and treatment in cerebral infarction patients.

Objective To establish a diagnosis method for fungal infection using two-round PCR,and to evaluate its sensitivity in the detection of clinical specimens suspected to be infected with fungi.Methods A total of 29 specimens of clinical sputum and alveolar wash solution were collected from patients with suspicious fungal infection.All specimens uaderwent direct microscopy with 10%KOH,fungal culture,one-round PCR and two-round PCR.The fungal universal primer targeting ITS regions of rDNA was used in PCR.The detection rate for fungi was compared between these methods.Results The detection rate for fungi was 20.69%by direct microscopy,37.9%by fungal culture,17.2%by one-round PCR,and 48.3%by two-round PCR.More than one species of fungus were detected in 6.9%(2/29),3.4%(1/29)and 24.1%(7/29)of these specimens by fungal culture.one-round PCR and two-round PCR, respectively.There was a significant difference in the detection rate between two-and one-round PCR(x2=6.34,P＜0.05).With regard to the detection rate for more than one species of fungus,two-round PCR was significantly higher than one-round PCR and fungal culture(x2=4.09,6.30.bom P＜0.05).Conclusion Two-round PCR may help to improve the sensitivity of molecular diagnosis of fungus-infected specimens.

Objective To report a case of vaginal colonization due to Trichosporon inkin. Methods A 34-year-old female presented with increased vaginal discharge accompanied by abnormal odor for 2 months. Clinical laboratory examination was carried out. Cultures of vaginal discharge yielded yeast-like colony. Subsequently, the isolate underwent the following mycological examinations: purification, slide micro-culture, temperature test, urea enzyme test, biochemistry identification, antifungal susceptibility test, and gene sequencing. Results Gynecological examination revealed white homogeneous secretions attached to mucous membrane of the vagina. Nugent scores of vaginal discharge amounted to 5-6. Two rounds of culture of vaginal discharge resulted in stramineous, reductus and yeast-like colony. The isolate could grow in 42 ℃. Appressorium on the top of hypha and typical sarcinae formed in slide microculture of corn agar, and yeast malt agar was the optimal growth medium for it. Urea enzyme test was positive. API 20C AUX biochemical test and gene sequencing revealed that the isolate was consistent with Trichosporon inkin. The isolate was sensitive to amphotericin B and azoles such as clotrimazole and fluconazole, but resistant to flucytosine and caspofungin. Conclusions It is the first report of vaginal colonization due to T. Inkin in China. The accu-rate identification of T. Inkin relies on synthetic analysis of phenotype characteristics, biochemistry test and molecular sequencing.

Objective To develop a PCR-RFLP method to rapidly identify filamentous fungi causing deep infection. Methods Universal fungal primers were used to amplify the internal transcribed spacer (ITS) region of Aspergillus fumigatus, Aspergillus Bavus, Aspergillus terreus, Aspergillus niger, Aspergillus versicolor, Aspergillus nidulans, Scedosporium apiospermum and Fusarium moniliforme followed by restriction fragment length polymorphism (RFLP) analysis with restrictive endonucleases Hha I, Hae III, Hinf I, Taq I and Msp I. Then, 22 clinical and 2 environmental fungal isolates were identified with the developed PCR-RFLP method. Results The RFLP analysis of PCR products with restrictive endonucleases Hha I and Hinf I allowed discrimination of 8 filamentous fungi causing invasive infection, and it took only 1 day to carry out the whole procedure from DNA extraction to PCR and restriction digestion. The identification results of 22 clinical strains and 2 environmental isolates with this PCR-RFLP method were completely consistent with those with conventional morphological method. Conclusion PCR-RFLP analysis is an efficient method for rapid identification of cultured filamentous fungi.

Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.

Objective To analyze the differential expression of Stathmin in human cells infected with human-tropic porcine endogenous retrovirus(PERV)and to explore the potential molecular effect of human-tropic PERV on human cells.Methods HEK293 cells were infected with the human-tropic PERV infectious molecular clone.PCR,real-time RT-PCR and immunofluorescence analysis were applied to confirm that HEK293 cells were infected.Then real-time RT-PCR and Western blot were carried out to analyze the differential expression of Stathmin at the mRNA level and protein level,respectively.Results HEK293 cells were infected by human-tropic PERV.Real-time RT-PCR and Western blot analysis showed that Stathmin was up-regulated in HEK293 cells infected with PERV compared with the control cells.Conclusion Stathmin was up-regulated in HEK293 cells infected with human-tropic PERV.These studies will be helpful for revealing the interaction of PERV and human cells,and for understanding the molecular effect of humantropic PERV on human cells.In addition,it suggested that PERV infection may infect cell growth and physiological functions,even be pathogenic.These will help to clarify the biologic characteristics of PERV and evaluate the safety of PERV in pig to human xenotransplantation.

Objective To genotype Trichosporon spp. with rDNA-ITSAGSl-RFLP analysis followed by cluster analysis, and attempt to apply this method to rapid species identification of human pathogenic Trichosporon spp.. Methods Fourteen strains of Trichosporon, which belonged to 8 species, were collected. The rDNA-ITS/IGSl regions were amplified by PCR and sequenced. Simultaneously, the amplicons were digested separately with restriction enzymes, including Hae III, Hha I , Hae IH and Hha I , Hinf I , Msp I and Taq I . Results The 8 species of Trichosporon could be classified into 4 subgroups with rDNA-ITS-RFLP, while inter-species identification of all the 14 strains from 8 species of Trichosporon could be realized with rDNA-IGSl-RFLP. Also, those genotypes of T. asahii which had relative long phylogenic distance could even be discriminated with rDNA-IGSl-RFLP. Conclusion The rDNA-ITS/IGSl-RFLP analysis is expected to be used in rapid interspecific identification of genus Trichosporon.

A 19-year-old man was admitted to the hospital for erythema and nodules on the face and postauricular region for 6 years. Microscopic examination of lesion scrapings revealed brown septate hyphae. A restricted, velvety and black colony grew on Sabouraud's dextrose agar. Slide culture on potato dextrose agar plate showed flask-shaped phialides at the apex of or around the hyphae with clear collarettes and flaring apex,mucilage-encapsuled, round to oval, semi-endogenous phialosporae accumulating at the apex of the phialides,giving a flower-like appearance. Anti-fungal susceptibility test showed that the fungus was sensitive to itraconazole, terbinafine and amphotericin B, but resistant to fluconazole. Sequence analysis of the ITS1-ITS4 region revealed a 98% consistency with the reference sequence of ITS1-ITS4 of Phialophora verrucosa. On the basis of above findings, the patient was diagnosed with cutaneous phaeohyphomycosis. Clinical improvement was seen after treatment with oral itraconazole (400 mg/d).

Objective To study the immune regulation of Galectin-9 on the active CD4+T cells and demonstrate the mechanisms.Methods Lymphocytes were harvested from wild-type C57BL/6 mouse,from which na(i)ve CD4+T cells were separated via MACS and then stimulated with anti-CD3 antibody(Ab) (2.5 μg/ml),anti-CD28 Ab(5 μg/ml) and IL-2(100 ng/ml) for 3 days.The active CD4+T cells were divided into 3 groups:Control group,Galectin-9 group and Galectin-9+α-lactose group.We detected the cell proliferation level by CFSE fluorescence intensity and then dynamically observed the cell morphological changes.The proportion of CD4+CD69+T cell,Th1,Th2 and Th17 cell was valued; Meanwhile,ELISA was used to detect the cytokine levels of IFN-γ,IL-4,IL-10,IL-12,IL-17A and TGF-β1 secreted by lymphocytes.Also Western blot was used to observe the changes of T cell differentiation regulatory protein such as T-bet,GATA-3 and ROR-γt.Results Compared with control group and Galectin-9+α-lactose group,in Galectin-9 group,the cell morphology began to change at 2 h.Moreover the proportions of CD4+CD69+ T cell,Th1 and Th17 cells decreased (P＜0.05),but no significant differences in Th2 cells.The level of IFN-γ,IL-12,IL-17A and TGF-β1 from the supernatant decreased (P＜0.05),while Th2-type cytokines IL-4 and IL-1O did not change.In addition,the expressions of T-bet and ROR-γt were significantly down-regulated (P＜0.05).Conclusion Galectin-9 inhibited Th1 and Th17-type immune response,while had no effect on Th2-type immune response.The mechanism of the immune regulation may be related to affect the expression of Th1 and Th17 specific transcription factors at transcription level.