Molecular phylogeny and morphology reveal a new wood-rotting fungal species, Sistotrema yunnanense sp. nov. from the Yunnan-Guizhou Plateau.

Wood-rotting fungi are important components of woody plant ecosystems and play an active role in the decomposition and turnover of nutrients from wood, and are among the major groups of Basidiomycota. In this study, a new species of wood-rotting fungus, Sistotrema yunnanense, was proposed based on morphological characteristics and molecular evidence. It is characterized by resupinate basidiomata, a monomitic hyphal system having generative hyphae with clamp connections, suburniform to urniform basidia, and short-cylindrical to oblong ellipsoid basidiospores (4.5-6.5 × 3-4 µm). Phylogenetic analyses performed using the large subunit nuc rDNA indicated that S. yunnanense was nested within the genus Sistotrema s.l. of the family Hydnaceae, within the order Cantharellales.

Exome sequencing identified a missense mutation of EPS8L3 in Marie Unna hereditary hypotrichosis.

BACKGROUND: Marie Unna hereditary hypotrichosis (MUHH) is an autosomal dominant disorder characterised by coarse, wiry, twisted hair developed in early childhood and subsequent progressive hair loss. MUHH is a genetically heterogeneous disorder. No gene in 1p21.1-1q21.3 region responsible for MUHH has been identified. METHODS: Exome sequencing was performed on two affected subjects, who had normal vertex hair and modest alopecia, and one unaffected individual from a four-generation MUHH family of which our previous linkage study mapped the MUHH locus on chromosome 1p21.1-1q21.3. RESULTS: We identified a missense mutation in EPS8L3 (NM_024526.3: exon2: c.22G->A:p.Ala8Thr) within 1p21.1-1q21.3. Sanger sequencing confirmed the cosegregation of this mutation with the disease phenotype in the family by demonstrating the presence of the heterozygous mutation in all the eight affected and absence in all the seven unaffected individuals. This mutation was found to be absent in 676 unrelated healthy controls and 781 patients of other disease from another unpublished project of our group. CONCLUSIONS: Taken together, our results suggest that EPS8L3 is a causative gene for MUHH, which was helpful for advancing us on understanding of the pathogenesis of MUHH. Our study also has further demonstrated the effectiveness of combining exome sequencing with linkage information for identifying Mendelian disease genes.

rhPLD2 suppresses chronic inflammation reactions in a guinea pig asthma model.

BACKGROUND: Asthma is a complex inflammatory disorder of the airways, and research on alternative therapeutic strategies has attracted attention. This study aimed at hypersusceptibility and toxicity of recombinant human phospholipase D2 (rhPLD2) in guinea pigs. We determined the behavioral responses in the model of immediate hypersensitivity animals and changes of eosinophil levels following use of the drugs. Special attention was given to the effects of rhPLD2 in vivo on the guinea pig with chronic persistent asthma and the mechanism involved. METHODS: To investigate the effect of rhPLD2 on the expression of protein kinase C (PKC), and to examine the activity of signal transducer and activator of transcription 1 and 5a in the lung of the guinea pig with chronic asthma. Guinea pigs with chronic asthma were divided into five groups: a saline group, a dexamethasone 5.0 mg group, and rhPLD2 (1.5, 2, or 3 mg) groups. Non-sensitized animals were as normal control group. PKC expression was measured by immunohistochemistry, alterations of STAT1 and STAT5a were detected by TransAM transcription factor assay kits. RESULTS: rhPLD2 (3.0 mg) decreased PKC expression to baseline and inhibited STAT1 activity compared with that of the saline group (p < 0.01). CONCLUSION: The rhPLD2 may suppress the chronic inflammatory reaction through down-regulating PKC expression and STAT1/STAT5a activity in the lung. The rhPLD2 may be a suitable therapeutic target for asthma.

Association of HLA haplotype with keloids in Chinese Hans.

Keloids are common abnormal raised fibroproliferative lesions that can occur following even minor cutaneous trauma. Human leukocyte antigen (HLA) polymorphisms have shown strong association with susceptibility to keloids with different ethnic backgrounds. In this study, the polymerase chain reaction sequence-specific primer method was used to analyze the distribution of HLA haplotype in 192 patients with keloids and 252 healthy control individuals. Controls were matched by gender, age, and race. We compared haplotype between the two groups, and analyzed their association with keloids. The haplotype analysis revealed that three new two-locus haplotypes including B*07-DQB1*0501, B*07-DRB1*15, DQB1*0503-DRB1*15 (P<0.05) were associated with keloids, while two extended haplotypes B*07-Cw*0802-DQB1*0501 (P=0.0063) and Cw*0802-DQB1*0501-DRB1*15 (P=0.0121) were found to be related to keloids. This is the first detailed report to elucidate HLA haplotypes associated with keloids. Our results provide some information for future research on predisposing genes in major histocompatibility complex (MHC) regions in Chinese patients with keloids. In addition, the association of certain HLA haplotypes with susceptibility to keloids would provide clues in choosing proper preventive strategies.

The association of the BLK gene with SLE was replicated in Chinese Han.

Systemic lupus erythematosus (SLE) is an autoimmune disease influenced by genetic and environmental factors. Recently, single nucleotide polymorphisms (SNPs) in the region of B lymphoid tyrosine kinase (BLK) have been shown to be associated with SLE in Caucasian population. In this paper, we genotyped SNP rs2248932 in 1,396 SLE patients of Chinese Han and 4,362 ethnically matched control subjects by using the Sequenom MassArray system. We confirmed that SNP rs2248932 in BLK gene was significantly associated with SLE (P = 1.41 x 10(-8) for the allele frequency, Odds ratio [OR] = 0.74, 95% confidence interval (CI): 0.66-0.82).The association of BLK in Chinese SLE patients was consistent with a dominant model (P = 8.88 x 10(-9), OR = 0.69, 95% CI: 0.61-0.79). In contrast to the Caucasian, this risk allele was the major allele in the Chinese Han; the risk allele frequency was higher in Chinese Han than in Caucasian. We did not find the association between this SNP and any subphenotype of SLE. The SNP rs2248932 was correlated to the expression of BLK mRNA. We conclude that the association of the BLK region with SLE was replicated in Chinese Han population living in mainland.

A single-nucleotide polymorphism of the TNFAIP3 gene is associated with systemic lupus erythematosus in Chinese Han population.

Systemic lupus erythematosus (SLE) is a complex systemic disease influenced by genetic and environmental factors. The exact pathogenesis of SLE is still unknown. Recently, several genome-wide association studies (GWA) in European population have found many novel susceptibility genes for SLE including TNFAIP3. In order to examine whether TNFAIP3 is associated with SLE in Chinese Han population, we genotyped one of its non-synonymous mutation SNP rs2230926, showing significant association evidence with SLE in European population, with 1,420 cases and 4,461 controls of Chinese Han by using Sequenom MassArray system. Highly significant association between SNP rs2230926 and SLE of Chinese Han was detected [OR = 1.65, 95% confidence interval (CI): 1.392-1.986, P = 2.03 x 10(-8)]. Interestingly, rs2230926 of TNFAIP3 was also associated with arthritis, ANA and some other subphenotypes of the disease. Our findings suggest that SNP rs2230926 in the TNFAIP3 might be a common genetic factor for SLE within different populations in terms of Chinese Han and European population.

Association of HLA class I alleles with keloids in Chinese Han individuals.

Keloids are benign fibroproliferative dermal tumors of unknown etiology. Some studies have suggested that human HLA status might potentiate development of keloids phenotype. No report has been published about HLA class I alleles associated with keloids in Chinese Han individuals. To investigate the etiology of keloids, the polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA class I alleles in 192 patients with keloids and 252 healthy controls in a Chinese Han population. The frequencies of HLA-A*03 (6.77% vs 0%, p(c) < 10(-7)), A*25 (10.16% vs 4.56%, p(c) = 0.0111), B*07 (7.81% vs 2.58%, p(c) = 0.0080), and Cw*0802 (19.79% vs 10.32%, p(c) = 0.0004) were significantly increased in keloid patients, whereas the frequency of HLA-A*01 (18.75% vs 38.10%, p(c) < 10(-7)) was highly decreased, compared with that in healthy controls. The A*03-B*07, A*25-B*07, A*03-Cw*0802, A*25-Cw*0802, and B*07-Cw*0802 were found as high-risk haplotypes in developing keloids in this study. No extended haplotype was found to be significantly related to keloids. Through stratified analysis, the association of subgroups (single site/multiple site, severity, and family history) of keloid patients with specific HLA alleles was identified. Our data suggest these alleles may be keloids susceptibility genes or may be in close linkage with the susceptibility genes.

[Roles of vascular endothelial growth factor and platelet-derived growth factor in lymphangiogenesis in epithelial ovarian carcinoma.].

OBJECTIVE: To assess roles of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in the mechanisms of lymphangiogenesis in epithelial ovarian carcinoma. METHODS: (1) Expression of Prox1, a newly described lymphatic endothelial cell nucleus marker, VEGF-A, VEGF-C, VEGF-D and PDGF-A, PDGF-B, PDGF-C, PDGF-D were detected by RT-PCR in SKOV3 cell line and in 90 ovarian tissue samples, included 15 benign tumors, 10 borderline tumors, 45 malignant tumors and 20 normal ovarian samples. (2) Expression levels of Prox1, VEGF-A, -C, -D and PDGF-A, -B, -C, -D were detected in 90 ovarian tissue sample mentioned above by real-time quantitative PCR (RTQ-PCR). RESULTS: (1) Prox1 was expressed in ovarian samples mentioned above, while not detected in SKOV3 cell. VEGF-A, -C, -D and PDGF-A, -B, -C, -D were found in SKOV3 cell and various ovarian tissues. (2) Expression levels of Prox1 (2.2 +/- 1.3, P < 0.01), VEGF-A (3.5 +/- 1.5, P < 0.01), VEGF-C (19 +/- 14, P < 0.01), VEGF-D (3.0 +/- 1.8, P < 0.01) and PDGF-A (3.3 +/- 3.3, P < 0.05), PDGF-C (6.9 +/- 4.6, P < 0.01) in malignant group were found to be significantly higher than those in borderline group and benign group. (3) The expression levels of Prox1, VEGF-A and PDGF-A were significantly greater in samples from the patients with lymph node metastasis (Prox1: 3.0 +/- 1.4, VEGF-A: 4.1 +/- 1.7, PDGF-A: 4.9 +/- 4.1), peritoneum metastasis (Prox1: 2.8 +/- 0.9, VEGF-A: 4.0 +/- 1.8, PDGF-A: 4.5 +/- 4.0) and in stage III - IV (Prox1: 2.6 +/- 1.3, VEGF-A: 4.0 +/- 1.4, PDGF-A: 4.1 +/- 3.7) than those without lymph node metastasis, without peritoneum metastasis and in stage I - II. There was a significant increased in the degree of VEGF-C and VEGF-D expression in positive lymph node metastasis group (VEGF-C: 24 +/- 13, VEGF-D: 3.9 +/- 2.0) compared with negative group (P < 0.05). (4) There were significant positive correlations between the expression levels of Prox1 and VEGF-D (r = 0.62, P < 0.01), PDGF-C (r = 0.91, P < 0.01) or PDGF-D (r = 0.61, P < 0.01). CONCLUSIONS: VEGF-A, VEGF-C and PDGF-A may promote lymphatic metastasis in epithelial ovarian carcinoma through else mechanisms other than lymphangiogenesis. VEGF-D may facilitate lymphangiogenesis and lymph node metastasis in epithelial ovarian cancer. There is no significant correlation between the expression of PDGF-B and lymphangiogenesis and lymph node metastasis. PCGF-C and PDGF-D may motivate lymphangiogenesis, but could not participate in lymph node metastasis in ovarian carcinoma.

Association of HLA-DQA1 and DQB1 alleles with keloids in Chinese Hans.

BACKGROUND: Some studies have suggested that human HLA status might potentiate development of keloids phenotype, and exists ethnic differences. No report has been published about HLA-DQA1 and DQB1 alleles associated with keloids in Chinese Hans. OBJECTIVES: To investigate whether HLA-DQA1 and DQB1 alleles are associated with genetic susceptibility to keloids in Chinese Hans. METHODS: Polymerase chain reaction-sequence-specific primer (PCR-SSP) method was used to analyze the distribution of HLA-DQA1 and DQB1 alleles among 192 patients with keloids and 273 healthy controls in Chinese Hans. RESULTS: (1) The frequencies of HLA-DQA1*0104, DQB1*0501 and DQB1*0503 (OR = 2.13, P(c) = 0.0063; OR = 14.42, P(c) < 10(-7) and OR = 6.09, P(c) < 10(-7), respectively) were significantly higher, while the frequencies of DQA1*0501, DQB1*0201 and DQB1*0402 (OR = 0.46, P(c) = 0.0099; OR = 0.24, P(c) < 10(-4) and OR = 0.10, P(c)=0.0054, respectively) were lower in patients than in controls. (2) In this study significant susceptibility haplotypes to keloids were DQA1*0104-DQB1*0501 and DQA1*0104-DQB1*0503. (3) HLA-DQB1*0501 and DQB1*0503 were positively associated with all subgroups of keloid patients. However, the DQA1*0104 (OR = 2.51, P(c) = 0.0009; OR = 2.22, P(c) = 0.0090 and OR = 2.20, P(c) = 0.0117, respectively) was only prevalent in keloid patients with single site, moderate severity and negative family history. (4) HLA-DQB1*0201 (OR = 0.27, P(c) = 0.0018 and OR = 0.27, P(c) = 0.0012, respectively) and DQB1*0402 (OR = 0.07, P(c) = 0.0270 and OR = 0.07, P(c) = 0.0306, respectively) were negatively associated with moderate severity and negative family history in keloids, moreover, HLA-DQB1*0201 (OR = 0.23, P(c) = 0.0003) and DQA1*0501 (OR = 0.43, P(c) = 0.0234) were less prevalent in patients with single site. CONCLUSION: This study demonstrated the positive association of HLA-DQA1 and DQB1 alleles and haplotypes with keloids.

[Study of bcl-2 antisense oligodeoxynucleotides on reversal of cisplatin resistance in ovarian cancer cell line A2780/DDP].

OBJECTIVE: To evaluate the influence of bcl-2 antisense oligodeoxynucleotides (AODN) on reversing cisplatin (DDP) resistance in ovarian carcinoma cell lines. METHODS: In the experiment groups, cisplatin resistant ovarian carcinoma cell line A2780/DDP cells were treated with cisplatin after oligodeoxynucleotides were transfected by lipofectamine. Cell proliferation was determined by counting the number of the cells continuously for six days. The expression level of bcl-2 mRNA was detected with reverse transcription polymerase chain reaction (RT-PCR) and protein was tested by immunofluorescence technique. The apoptosis of ovarian cancer cell lines was measured by DNA ladder assay. The inhibitory rate of cells was detected by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT). The cells treated with culture media were used as the negative control, and the untreated cells were used as the blank. RESULTS: The proliferation of AODN transfected cells was inhibited significantly compared with that in the control groups, and the logarithmic growth delayed for 24h. The expression of bcl-2 mRNA and bcl-2 protein in AODN transfected cells was 0.76 +/- 0.01 and 171.3 +/- 3.1 respectively. Compared with the expression of bcl-2 mRNA 1.56 +/- 0.03 and bcl-2 protein 105.3 +/- 1.7 in the blank, the difference was significant (P < 0.01), but compared with the same concentration of sense oligodeoxynucleotides (SODN) or random-oligodeoxynucleotides (RODN) transfected cells, the difference was not significant (P > 0.05). The AODN transfected cells were sensitive to DDP again, and the inhibitory rate was (97.49 +/- 1.00)%, which was higher than that of the negative control (P < 0.01). In the cells treated with 16 microg/ml AODN for 72 h followed by DDP, DNA ladder was obviously observed with agarose gel electrophoresis, but in the RODN and SODN transfected cells, DNA ladder could not be seen. CONCLUSION: Through inhibiting the expression of bcl-2 mRNA and protein and increasing the apoptosis, AODN can reverse the resistance to cisplatin in ovarian cancer cell line A2780/DDP.

[Killing effect of adenovirus mediated fusion gene cytosine and deaminase uracil phosphoribosyl transferase directed by glutathione S-transferase P1 promoter on cisplatin-resistant ovarian cancer cells in vitro].

OBJECTIVE: To construct an adenoviral vector in which the fusion gene cytosine and uracil phosphoribosyl (UPP) transferase was directed by glutathione S-transferase P1 (GSTP1) promoter, and to investigate specific killing effect of the suicide gene system on cisplatin-resistant ovarian cancer cells. METHODS: Recombinant adenovirus was generated through homologous recombination in bacteria. A2780 and A2780/DDP cells were infected with Ad and then received flucytosine (5-FC) administration. The relative survival of these cells was tested. And a bystander effect was observed by mixing gene-transferred and gene-untransferred A2780/DDP cells with 5-FC. RESULTS: In vitro, when MOI was 100 and 5-FC was 250 micro g/ml, relative survival rate of A2780/DDP cells was only (3.6 +/- 1.0)%; that of A2780 cells was (76.5 +/- 2.8)%. Significant bystander effect was caused by CD-UPP gene and 20% gene-transferred A2780/DDP cells induced 80.3% of total cells to death. CONCLUSION: Recombinant adenovirus carrying CD-UPP gene driven by GSTP1 promoter has a specific killing effect on cisplatin-resistant ovarian cancer cells.