Diversity of porcine reproductive and respiratory syndrome virus in Shandong, China.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically significant pathogens in swine industry of China. To study infection and genetic variation of PRRSV, 637 tissue samples were collected from diseased pigs in Shandong, and then subjected to detection of PRRSV. The nsp2 and ORF5 genes were sequenced for investigation of variations and phylogenetic analysis. The results showed that positive rate of PRRSV was 9.58% in the collected samples. Phylogenetic analysis of GP5 showed that these strains were clustered into two lineages (1 and 8) indicating different genotypes of PRRSV were circulating in Shandong province. Meanwhile, sequence analysis Of nsp2 showed that the PRRSV strains with 30 amino acids deletions were dominant. Moreover, novel pattern of recombination/deletion and insertion in nsp2 was observed in these strains, indicating that novel PRRSV strains with different patterns of deletions or insertions in nsp2 are emerging in China. All the results suggested that continuous surveillance of PRRSV in China is warranted. Keywords: PRRSV; GP5; nsp2; genetic analysis; Shandong.

Genetic characterization and mutation analysis of Qihe547 Aujeszky's disease virus in China.

BACKGROUND: Aujeszky's disease virus (ADV) can cause neurologic disease in young pigs, respiratory disease in older pigs and abortion or birth of mummified fetuses or stillborn neonates. The re-emergence of Aujeszky's disease (AD) in pig farms vaccinated with live vaccine (Bartha-K61) caused substantial economic losses to Chinese pig industry since late 2011. A field ADV, named Qihe547, was isolated from pigs that exhibited suspected AD clinical symptoms. To better understand the genetic characteristics and mutations of Qihe547 ADV, the whole genome was sequenced and analyzed. RESULTS: The genomic length of Qihe547 ADV was 143,404 bp, with 73.59% G + C contents. Phylogenetic analysis based on the whole genome of ADV strains revealed that Chinese ADV strains were located to one group with three subgroups. Qihe547 ADV was closely related to these novel ADV strains isolated in China since 2012. Qihe547 presented numerous hypervariable regions compared with oversea ADV strains. In 34 genes of Qihe547 ADV, amino acid (AA) insertion or deletion were observed. In addition, numerous AA mutations were found in the main protective antigen genes (gB, gC and gD genes). The differences of potential antigenic peptides in the main protective antigens between Qihe547 ADV and ADV Bartha were discovered in the dominant antigenic regions of gB (AA59-AA126, AA507-AA734),the extracellular region of gC and gD. CONCLUSION: High diversity was observed between Qihe547 and foreign ADV isolates. The AA variations and the differences of potential antigenic peptides in the important functional regions of the main protective antigen (gB, gC and gD) of ADV Qihe547 may contribute to immune evasion of the virus and may be partial reason that the virus escapes from the vaccination of Bartha-K61 vaccine. In a word, the effect of the variations obviously requires further research.

Multiple amino acid transporters as carriers load L-valine-phenazine-1-carboxylic acid conjugate into Ricinus sieve tubes for the phloem translocation.

Some transporters play important roles in the uptake and acropetal xylem translocation of vectorized agrochemicals. However, it is poorly understood the basipetally phloem-loading functions of transporters toward vectorized agrochemicals. Here, L-Val-PCA (L-valine-phenazine-1-carboxylic acid conjugate) uptake was demonstrated carrier-mediated. RcAAP2, RcANT7, and RcLHT1 showed a similarly up-regulated expression pattern from 62 transporter coding genes in Ricinus at 1 h after L-Val or L-Val-PCA treatment. Subcellular localization revealed that fusion RcAAP2-eGFP, RcANT7-eGFP and RcLHT1-eGFP proteins were expressed in the plasma membrane of mesophyll and phloem cells. Yeast assays found that RcAAP2, RcANT7, and RcLHT1 facilitated L-Val-PCA uptake. To further demonstrate the phloem-loading functions, using vacuum infiltration strategy, an Agrobacterium-mediated RNA interference (RNAi) protocol was constructed in seedlings. HPLC detection indicated that L-Val-PCA phloem sap concentrations were significantly decreased 54.5 %, 27.6 %, and 41.6 % after silencing for 72 h and increased 48.3 %, 52.6 %, and 52.4 % after overexpression, respectively. In conclusion, the plasma membrane-located RcAAP2, RcANT7, and RcLHT1 can loaded L-Val-PCA into Ricinus sieve tubes for the phloem translocation, which may aid in the utilization of transporters and molecular design of phloem-mobile fungicides target root or vascular pathogens.

[Value of MiR-155 in Prognostic Evaluation of Elderly Patients with Primary Immune Thrombocytopenia].

OBJECTIVE: To explore the value of has-microRNA-155 (miR-155) in peripheral blood mononuclear cells (PBMNC) in prognostic evaluation of elderly patients with primary immune thrombocytopenia (PITP). METHODS: One hundred and thirty elderly PITP patients and 60 healthy volunteers in our hospital were selected. The relative expression level of miR-155 in PBMNC was detected by RT-PCR. Unconditional logistic multivariate regression analysis was used to analyze the relationship between miR-155 expression and prognosis of PITP patients, and Kaplan-Meier was further used to analyze the relationship between miR-155 and PITP recurrence. RESULTS: The relative expression level of miR-155 in PBMNC of elderly PITP patients was significantly higher than that in healthy volunteers, and increased significantly with the severity of the disease (P＜0.05). The overall effective rate of elderly PITP patients with miR-155 low-expression was significantly higher than that in the patients with miR-155 high-expression (96.92% vs 72.31%) by after treatment with glucocorticoid. Multivariate analysis showed that miR-155 was an independent risk factor for PITP patients. Elderly patients with high expression of miR-155 showed a higher risk of recurrence. CONCLUSION: miR-155 in PBMNC has a high accuracy for PITP diagnosis, and the elderly patients with high level of miR-155 show a poor prognosis and a higher risk of recurrence.

[Effect of MDR1 and CYP3A5 gene polymorphisms on outcomes of patients receiving imatinib treatment for chronic myeloid leukemia].

OBJECTIVE: To study the effect of MDR1 and CYP3A5 gene polymorphisms on the outcomes of imatinib treatment in patients with chronic myeloid leukemia (CML). METHODS: A total of 100 patients with CML treated with imatinib were enrolled in this study, including 50 patients with cytogenetic relapse (study group) and 50 without cytogenetic relapse (control group) during the follow-up for 45 months. For all the patients, single nucleotide polymorphisms (SNPs) of C1236T, C3435T, and G2677T/A loci in the MDR1 gene and A6986G locus in CYP3A5 gene were genotyped and the trough levels of imatinib was measured using LC-MS/MS. The relationship between SNPs of the loci and the risk of cytogenetic relapse were analyzed. RESULTS: The risk of cytogenetic recurrence was significantly higher in patients with CC genotypes of MDR1-C1236T and MDR1-C3435T than in those with CT + TT genotypes (P < 0.05). The median survival time of the patients with TT genotypes of MDR1-C3435T and MDR1-C1236T was significantly higher than that of patients with CC genotypes and CT genotypes (P < 0.05). The incidences of hematologic toxicity and neutropenia were significantly higher in patients with cytogenetic relapse than in those without cytogenetic relapse (P < 0.05). MDR1-C3435T genotype and imatinib concentration were independent predictors of cytogenetic relapse of CML. CONCLUSIONS: The risk of cytogenetic relapse of CML was significantly affected by SNPs of C1236T and C3435T loci of MDR1 gene and blood imatinib concentration. MDR1-C3435T genotype can be used as a potential biomarker for predicting cytogenetic relapse in CML patients.

[Heavy Metal Contamination of Soils and Crops near a Zinc Smelter].

To evaluate the influence of mining activities on heavy metal contamination and health risks in a zinc-smelting area in Guizhou Province, Pb, Cd, Zn, and Cu in agriculture soils and dominant crops were monitored and analyzed. A single factor pollution index and a comprehensive pollution index were calculated to assess the quality of soils and crops. Furthermore, the potential ecological risks of the four heavy metals in soils and crops were evaluated using the potential ecological risk index (RI). The potential health risks to adults and children induced by the intake of these heavy metals through consumption of crops were evaluated using the hazard quotient (HQ). The results showed that:①The farmlands in the zinc-smelting area were contaminated by heavy metals; the most polluted farm land was corn fields followed by paddy fields and wheat fields. The order of the heavy metal concentrations was Cd > Cu > Zn > Pb. The Nemero index of corn fields was higher than the threshold value of heavy pollution and was higher than that of the paddy and wheat fields. Cd contributed most to the integrated pollution index. ②The very high ecological risk was posed by Cd followed by Pb, Cu, and Zn. The results of the potential ecological risk assessment showed that 1.41% of the sites were classified as low risk, 21.1% classified as medium risk, 35.2% as high risk, and 42.3% as extremely high risk. ③The average concentrations of Pb, Cd, Zn, and Cu in rice were 0.145, 0.017, 16.97, and 2.704 mg·kg-1, respectively. The average concentrations of Pb, Cd, Zn, and Cu in corn were 0.094, 0.055, 26.81, and 4.464 mg·kg-1, respectively. The average concentrations of Pb, Cd, Zn, and Cu in wheat were 0.048, 0.085, 35.37, and 5.426 mg·kg-1, respectively.④Rice, corn, and wheat from the study area were subjected to pollution at different degrees. Among the three crops studied, wheat was polluted most heavily, followed by rice and corn. Contamination levels of rice and corn were safe, whereas the contamination of wheat reached alarming levels. ⑤The daily intake of individual heavy metals from food was below the standard exposure dose set by the United States Environmental Protection Agency. In addition, the assessment results for the comprehensive health risk index for the three crops indicate that the contamination of heavy metals poses health risks to adults and children by food ingestion. ⑥ Positive correlations were not observed between farmlands and crops.