Analysis of Pyricularia oryzae and P. grisea from Different Hosts Based on Multilocus Phylogeny and Pathogenicity Associated with Host Preference in China.

Pyricularia oryzae and P. grisea are important agents of major diseases on a wide range of gramineous hosts. Whereas P. oryzae is the most important pathogen causing outbreaks of rice blast, P. grisea is mainly a pathogen of crabgrass. In this study, 103 P. oryzae and 20 P. grisea isolates were collected from seven species of plants, and we analyzed their phylogeny, pathogenicity, and relationship with host preferences to investigate the differences among them from different hosts. Based on phylogenetic analysis of multilocus sequences, 16 isolates from crabgrass and four isolates from green bristlegrass were identified as P. grisea and another 103 isolates from crabgrass, green bristlegrass, goose grass, foxtail millet, wild millet, rice, and sedge belonged to P. oryzae. Results of pathogenicity tests by artificial inoculation demonstrated that six of 10 P. oryzae isolates from rice and three of 44 P. oryzae isolates from green bristlegrass showed cross-infectivity on green bristlegrass and rice, respectively. Taken together, our results demonstrated that isolates from green bristlegrass and crabgrass consist of both P. oryzae and P. grisea and that P. oryzae isolates showed cross-infectivity between rice and green bristlegrass, suggesting that host shifts may occur for P. oryzae and P. grisea.

Methylation in the promoter regions of WT1, NKX6-1 and DBC1 genes in cervical cancer tissues of Uygur women in Xinjiang.

This study aimed to explore: 1) DNA methylation in the promoter regions of Wilms tumor gene 1 (WT1), NK6 transcription factor related locus 1 gene (NKX6-1) and Deleted in bladder cancer 1 (DBC1) gene in cervical cancer tissues of Uygur women in Xinjiang, and 2) the correlation of gene methylation with the infection of HPV16/18 viruses. We detected HPV16/18 infection in 43 normal cervical tissues, 30 cervical intraepithelial neoplasia lesions (CIN) and 48 cervical cancer tissues with polymerase chain reaction (PCR) method. Methylation in the promoter regions of the WT1, NKX6-1 and DBC1 genes in the above-mentioned tissues was measured by methylation-specific PCR (MSP) and cloning sequencing. The expression level of these three genes was measured by real-time PCR (qPCR) in 10 methylation-positive cervical cancer tissues and 10 methylation-negative normal cervical tissues. We found that the infection of HPV16 in normal cervical tissues, CIN and cervical cancer tissues was 14.0, 36.7 and 66.7%, respectively. The infection of HPV18 was 0, 6.7 and 10.4%, respectively. The methylation rates of WT1, NKX6-1 and DBC1 genes were 7.0, 11.6 and 23.3% in normal cervical tissues, 36.7, 46.7 and 30.0% in CIN tissues, and 89.6, 77.1 and 85.4% in cervical cancer tissues. Furthermore, WT1, NKX6-1 and DBC1 genes were hypermethylated in the high-grade squamous intraepithelial lesion (CIN2, CIN3) and in the cervical cancer tissues with infection of HPV16/18 (both P< 0.05). The expression of WT1, NKX6-1 and DBC1 was significantly lower in the methylation-positive cervical cancer tissues than in methylation-negative normal cervical tissues. Our findings indicated that methylation in the promoter regions of WT1, NKX6-1 and DBC1 is correlated with cervical cancer tumorigenesis in Uygur women. The infection of HPV16/18 might be correlated with methylation in these genes. Gene inactivation caused by methylation might be related to the incidence and development of cervical cancer.

High Expression of long non-coding RNA PVT1 predicts metastasis in Han and Uygur Patients with Gastric Cancer in Xinjiang, China.

To analyze the level and diagnostic value of plasmacytoma variant translocation 1 (PVT1) in gastric cancer (GC) of Han and Uygur in Xinjiang, China, we collected 42 GC and 47 normal gastric tissues and performed tissue microarray. In situ hybridization was used to detect PVT1, while immunohistochemistry was used to analyze c-myc. The relationship between PVT1, c-myc and clinical pathological features was investigated. We then analyzed the expression of PVT1 in six GC cell lines. RNA interference was used to silence PVT1 in BGC823 and AGS cells. c-myc was detected by western blotting after silencing PVT1, while proliferation, invasion and migration ability were also analyzed. We found that PVT1 and c-myc were highly expressed in both Han and Uygur GC tissues. In Han GC, PVT1 was correlated with lymph node metastasis and primary tumor site. In Uygur GC, both PVT1 and c-myc were correlated with lymph node metastasis and clinical staging. PVT1 was positively correlated with c-myc. BGC823 and AGS cells exhibited high levels of PVT1. When PVT1 expression was silenced, the expression of c-myc decreased, while migration and invasion ability were also decreased in cells. PVT1 could therefore be a potential biomarker to predict the metastatic tendency of GC in both Han and Uygur patients.

Expression levels and the prognostic value of long non-coding RNA PVT1 in serum of Han and Uygur gastric cancer patients in Xinjiang, China.

BACKGROUND: The expression level of a long non-coding RNA (lncRNA), plasmacytoma variant translocation 1 (PVT1), was studied in serum of Han and Uygur gastric cancer (GC) patients and normal persons in Xinjiang, China, and the prognostic value of PVT1 was analyzed. METHODS: We collected serum samples from 87 GC patients (51 Han and 36 Uygur), and 95 normal persons (55 Han and 40 Uygur). Total RNA was extracted from the serum and used for real-time polymerase chain reaction (PCR) to detect the level of PVT1.The relationships between PVT1 and clinicopathological features were analyzed using the Spearman's relative analysis, rank sum test, and χ2 test. At the same time, selection electrochemiluminescence was used to detect the concentration of serum tumor markers, including alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), and carbohydrate antigen 72-4 (CA72-4). The correlation between PVT1 and the tumor markers was analyzed by Spearman's relative analysis. RESULTS: The serum expression level of PVT1 was higher than that of the normal group in both Han and Uygur GC patients (P<0.05). PVT1 expression in Uygur GC serum was higher than that of Han patients (P<0.05). The high level of serum PVT1 correlated with lymph node metastasis in both Han and Uygur GC patients (P<0.05). Compared with Han GC patients, the Uygur people were more likely to develop distant metastasis (P<0.05). Uygur patients were more often diagnosed at stage III or IV (P<0.05). In addition, serum PVT1 showed a significant correlation with CA19-9 in Han GC patients (P<0.05). CONCLUSIONS: Increased serum PVT1 level may indicate an increased tendency for lymphatic metastasis, especially in Uygur GC patients. When combined with CA19-9, the PVT1 expression level may be a better diagnostic marker in Han GC patients.

Variants of human papillomaviruses 16 (HPV16) in Uigur women in Xinjiang, China.

BACKGROUND: Persistent infection of high-risk human papillomaviruses 16 (HPV16) has been considered as the leading cause of cervical cancer. In this study we assessed HPV16 sequence variation and genetic diversity of HPV16 variants in cervical cancer in Uigur women in Xinjiang, China. We analyzed the nucleotide sequences of the open reading frames of E6 and E7, and part of the open reading frames of L1 of HPV16 in Uigur women. METHODS: Biopsies of histologically confirmed HPV16 infections with cervical cancer were obtained from 43 Uigur women in Xinjiang, China. E6, E7 and L1 genes of HPV16 of all samples were amplified and sequenced; the sequences were used in phylogenetic analysis of HPV16 variants. RESULTS: Our analysis revealed nine nucleotide changes in E6 (five changes), E7 (one change) and L1 (three changes) gene. The most frequently observed variations were T350G (79.1 %). One variation T295G (D64E) at E6 were detected in 6 cases (KT959536, KT959542, KT959546, KT959550, KT959553, KT959558). Deletion (464Asp) along with insertion (448Ser) were observed in L1 (100 %). Most variants were European lineage (97.7 %); only one belongs to Asia variants with common T178G (D25E) in E6 and A647G (N29S) in E7. CONCLUSION: The most prevalent HPV16 variants in the Uigur women we studied were of the European lineage. Our results indicate that HPV16 European lineage may serve as a harmful factor associated with the development and progression of cervical cancer.

Methylation in the promoter regions of WT1, NKX6-1 and DBC1 genes in cervical cancer tissues of Uygur women in Xinjiang

Abstract This study aimed to explore: 1) DNA methylation in the promoter regions of Wilms tumor gene 1 (WT1), NK6 transcription factor related locus 1 gene (NKX6-1) and Deleted in bladder cancer 1 (DBC1) gene in cervical cancer tissues of Uygur women in Xinjiang, and 2) the correlation of gene methylation with the infection of HPV16/18 viruses. We detected HPV16/18 infection in 43 normal cervical tissues, 30 cervical intraepithelial neoplasia lesions (CIN) and 48 cervical cancer tissues with polymerase chain reaction (PCR) method. Methylation in the promoter regions of the WT1, NKX6-1 and DBC1 genes in the above-mentioned tissues was measured by methylation-specific PCR (MSP) and cloning sequencing. The expression level of these three genes was measured by real-time PCR (qPCR) in 10 methylation-positive cervical cancer tissues and 10 methylation-negative normal cervical tissues. We found that the infection of HPV16 in normal cervical tissues, CIN and cervical cancer tissues was 14.0, 36.7 and 66.7%, respectively. The infection of HPV18 was 0, 6.7 and 10.4%, respectively. The methylation rates of WT1, NKX6-1 and DBC1 genes were 7.0, 11.6 and 23.3% in normal cervical tissues, 36.7, 46.7 and 30.0% in CIN tissues, and 89.6, 77.1 and 85.4% in cervical cancer tissues. Furthermore, WT1, NKX6-1 and DBC1 genes were hypermethylated in the high-grade squamous intraepithelial lesion (CIN2, CIN3) and in the cervical cancer tissues with infection of HPV16/18 (both P< 0.05). The expression of WT1, NKX6-1 and DBC1 was significantly lower in the methylation-positive cervical cancer tissues than in methylation-negative normal cervical tissues. Our findings indicated that methylation in the promoter regions of WT1, NKX6-1 and DBC1 is correlated with cervical cancer tumorigenesis in Uygur women. The infection of HPV16/18 might be correlated with methylation in these genes. Gene inactivation caused by methylation might be related to the incidence and development of cervical cancer.