Objective To explore the feasibility of the nested PCR technique to detect the Trichosporon asahii DNA in the blood and liver samples from the murine model of disseminated trichosporonosis.Methods On day 3,7,14 after intravenous inoculation of Trichosporon asahii suspension,blood and liver samples of 30 BALB/c mice were collected.Another ten normal mice serve as control.DNA was amplified by nested PCR,with a pair of primer.Fungal culture was done as control.Results On day 3,7,14 after inoculation,the positive rate of the serum fungal cultivation were 0,20%,0;the positive rate of the serum after nested PCR assay was 80%,90%, 88.9%;the positive rate of liver fungal cultivation were 50%,40%,22.2%;the positive rate of liver after nested PCR assay was 90%,90%,88.9%.There were statistical differences between the positive rate by nested PCR assay and fungal cultivation to check the serum and liver samples on day 3,7,14 after inoculation.The survival mice to the 14~(th) day were too less to make statistical analysis.Conclusion The results suggest that T.asahii DNA assay by nested PCR is more specific,more sensitive,and faster than fungal cultivation.The nested PCR assay could be a potential assay for the diagnosis of disseminated trichosporonosis.

Objective To observe the changes of pulmonary function parameters in healthy middle-aged and elderly people in Shanghai.Methods A total of 298 subjects aged 45 years and over with normal pulmonary function and no smoking history were selected among subjects who underwent routine physical examination from Jun 2009 to Sep 2011.The subjects were divided into three groups:45-59 years,60-69 years,and ≥70 years.The tested pulmonary function included 13 parameters:vital capacity (VC),residual volume (RV),functional residual capacity (FRC),total lung capacity (TLC),RV/TLC,forced vital capacity (FVC),forced expiratory volume in one second (FEV1),FEV1/FVC,peak expiratory flow (PEF),forced expiratory flow at 25 ％ of FVC exhaled (FEF25),forced expiratory flow at 50％ of FVC exhaled (FEF50),diffusion capacity of the lung for carbon monoxide (DLCO),and specific diffusion capacity of CO (KCO).Results Compared with the young adults,7 pulmonary function parameters including VC,FVC,FEV1,PEF,TLC,DLCO and KCO were declined significantly in healthy middle aged and elderly people Compared with the young adults,pulmonary function parameters including RV and RV/TLC were increased [male (1.9 ±0.4)Lvs.(1.6±0.3)L,(34.7±4.9)％ vs.(26.4±4.3)％,P＜0.01;female (1.5±0.3)L vs.(1.3±0.3)L,(36.1±5.3)％ vs.(30.3±5.5)％,P＜0.01].Although RVincreased with aging in middle-aged and elderly people,but there was no statistical significance (P＞0.05).FRC didn't change obviously.FEV1/FVC was lower in old people than in young people[male (80.7±4.3)％ vs.(84.2±5.6)％,P ＜0.01;female (82.5± 5.5) ％ vs.(86.4±4.8)％,P＜0.01],but the difference between 3 groups of middle-aged and the elderly was not significant.The FEV1/FVC of men and women aged 70 and more were (80.2±4.5)％ and (81.8±5.6)％,respectively.Conclusions Both ventilation function and diffusion function of healthy elderly people are decreased with ageing,RV is increased and FRC is steady,and the declining tendency of FEV1/FVC obviously slows down with aging in middle aged and elderly people.

Objective To screen, analyze and predict microRNAs (miRNAs) related to systemic scle-roderma (SSc). Methods Differentially expressed miRNAs between tissue samples from 3 patients with SSc and 3 normal human controls were screened with a gene chip including 924 miRNAs. Target genes regulated by differentially expressed miRNAs were searched with bioinformatics method. Finally, miRNAs related to SSc were predicted. Results There were 24 miRNAs differentially expressed between tissue samples of SSc and normal controls, including 9 up-regulated miRNAs and 15 down-regulated miRNAs. Literature review disclosed that SSc was associated with target genes regulated by hsa-miR-206, has-let-7g, hsa-miR-133a, hsa-miR-125b, hsa-miR-40-5p and hsa-miR-23b. In particular, 15 target genes regulated by hsa-miR-206 were closely correlated with the pathogenesis of SSc. Conclusions In lesions of SSc, there is an expression of miRNAs related to the pathogenesis of SSc, which may include hsa-miR-206 as well as 5 other miRNAs.

OBJECTIVE To observe the significance of(1,3)-?-D-glucan in different humor samples on the rat model of systemic trichosporosis.METHODS Established the animal model with systemic infection of trichosporosis,to collect the bronchoalveolar lavage fluid(BALF),plasma and urine samples for determining,and checked with G-test,nested PCR and fungus culture.RESULTS In the earlier infection(10d),the sensitivity of G-test in the samples of plasma,BALF and urine were 80.00%,86.67%,and 6.67%,respectively,and had significant difference with control group.In the 30 plasma samples,the average sensitivity of G-test and nested-PCR blood fungus culture was 76.76%,68.85%,and 3.33%,respectively.With the statistical analysis,the sensitivity of G-test had significant deviation than fungus culture(P

OBJECTIVE To identify if Trichosporon asahii exist cph1 gene homolog of Candida albicans,and analyze its nucleotide sequence.METHODS Nuclear DNA was extracted from the cells of T.asahii by using a simplified protocol,designed 29 pairs of primers according to the cph1 gene sequence of C.albicans and amplify by PCR.The PCR products were cloned and sequenced using the ABI377 nucleotide sequenator,and BLAST analysis.RESULTS An 827 bp gene was amplified successfully,which was homolog with the cph1 gene of C.albicans and their identity was 97.3%.CONCLUSIONS This trial determines the clone cph1 gene in T.asahii for the first time,which makes bases for the role of cph1 gene in the hypha formation.

Objective To explore the expression and significance of COX-2 ,MMP-9 and PTEN in colonic adenocarcinoma tissues .Methods Seventy-two cases of pathologically archived paraffin specimens after colonic adenocarcinoma radical operation in the hospital from March 2014 to May 2016 were collected ,30 cases of colonic adenoma tissues and 30 cases of normal mucosal mem-brane tissues served as the controls .The expressions of COX-2 ,MMP-9 and PTEN were detected by adopting the immunohisto-chemical SP method .Then the correlation between COX-2 ,MMP-9 and PTEN with clinicopathological features of colonic adenocar-cinoma was analyzed .Results The positive expression rate of COX-2 and MMP-9 in colonic adenocarcinoma tissues was significant-ly higher than that in colonic adenoma tissues and normal colon mucosal membrane tissues (P<0 .05);the positive expression rate of PTEN was significantly lower than that in colonic adenoma tissues and normal colon mucosal membrane tissues (P<0 .05) .The expression of COX-2 ,MMP-9 and PTEN in colonic adenocarcinoma was correlated with the tumor invasive depth ,lymph node me-tastasis and TNM staging(P<0 .05) ,while had no correlation with sex ,age ,tumor morphology ,tumor size and differentiation de-gree(P>0 .05) .COX-2 ,MMP-9 and PTEN expressions showed negative correlation (r= -0 .260 ,-0 .282 ,P<0 .05) ,COX-2 and MMP-9 expression showed a positive correlation (r=0 .335 ,P=0 .004) .Conclusion The abnormal expression and interaction of COX-2 ,MMP-9 and PTEN are closely correlated with the occurrence ,invasion and metastasis of colonic adenocarcinoma .Their combined detection has an important significance for early diagnosis and prognosis assessment of colonic adenocarcinoma .