Exosome-mediated transfer of MIF confers temozolomide resistance by regulating TIMP3/PI3K/AKT axis in gliomas.

Temozolomide (TMZ) resistance is an important cause of clinical treatment failure and poor prognosis in gliomas. Increasing evidence indicates that cancer-derived exosomes contribute to chemoresistance; however, the specific contribution of glioma-derived exosomes remains unclear. The aim of this study was to explore the role and underlying mechanisms of exosomal macrophage migration inhibitory factor (MIF) on TMZ resistance in gliomas. We first demonstrated that MIF was upregulated in the exosomes of TMZ-resistant cells, engendering the transfer of TMZ resistance to sensitive cells. Our results indicated that exosomal MIF conferred TMZ resistance to sensitive cells through the enhancement of cell proliferation and the repression of cell apoptosis upon TMZ exposure. MIF knockdown enhanced TMZ sensitivity in resistant glioma cells by upregulating Metalloproteinase Inhibitor 3 (TIMP3) and subsequently suppressing the PI3K/AKT signaling pathway. Additionally, exosomal MIF promoted tumor growth and TMZ resistance of glioma cells in vivo, while IOS-1 (MIF inhibitor) promotes glioma TMZ sensitive in vivo. Taken together, our study demonstrated that exosome-mediated transfer of MIF enhanced TMZ resistance in glioma through downregulating TIMP3 and further activating the PI3K/AKT signaling pathway, highlighting a prognostic biomarker and promising therapeutic target for TMZ treatment in gliomas.

Evaluation of Potato virus X mild mutants for cross protection against severe infection in China.

BACKGROUND: Cross protection is a promising alternative to control plant viral diseases. One critical factor limiting the application of cross protection is the availability of attenuated mutants or mild strains. Potato virus X (PVX) infects many crops and induces huge economic losses to agricultural production. However, researches on the variability and mechanism of PVX virulence are scarce. METHODS: The mutants were obtained by introducing mutations into the RNA dependent RNA polymerase (RdRp) gene of PVX via site-directed mutagenesis. Attenuated mutants were screen according to their symptoms in Nicotiana benthamiana plants. The protection efficacy against severe infection were evaluated with interval of 5, 10 and 15 days. RESULTS: Among the 40 mutants obtained, four mutants carrying substitutions of either Glu46, Asn863, Asn968 or Glu1001 to Ala in PVX RdRp showed drastically attenuated symptom, accompanying with reduced accumulation levels of coat protein, plus- and minus-sense RNAs. When the interval between protective and challenging inoculations was 15 days, mutant E1001A (with substitution of Glu1001 to Ala in RdRp) provided complete protection against severe infection in both Nicotiana benthamiana and tomato, while E46A (Glu46 mutated to Ala) provided incomplete protection. To reduce the risk of reverse mutation, we constructed mutant dM which carries double mutations of both Glu46 and Glu1001 to Ala in RdRp. The mutant dM could provide effective protection against severe PVX infection. CONCLUSION: Mutations of Glu46, Asn863, Asn968 or Glu1001 to Ala in PVX RdRp significantly reduced the viral symptoms. Mutants E1001A and E46A could provide effective protection against wild type PVX in both Nicotiana benthamiana and tomato. These results provide theoretical and practical bases for the control of PVX via cross protection.

Molecular characterization of ascaris from Tibetan pigs by three mitochondrial markers of nad1, cox1 and cox2.

Ascaris is a helminthic parasite, which infects a wide range of host species causing ascariasis, a predominant disease worldwide. This parasite causes significant economic losses to the pig industry. The current study was designed to determine the Ascaris nematode by the genetic characterization of three mitochondrial (mt) genes namely NADH dehydrogenase subunit 1 (nad1), cytochrome oxidase subunit 1 (cox1) and cytochrome oxidase subunit 2 (cox2). A high infection rate of Ascaris nematode has been found in Tibetan pigs at the slaughter houses in Tibet Autonomous Region of China. The nad1, cox1 and cox2 genes sequences collected from adult Ascaris individuals were amplified by polymerase chain reaction. The cloned-amplicons and the positive products were sequenced and phylogenetic analysis was performed. The results indicated that the Ascaris infecting the Tibetan pigs were Ascaris suum (A. suum). This is the first report on the isolation, identification and genetic characterization of three mitochondrial genomes (nad1, cox1, and cox2) of A. suum originated from Tibetan pigs at high altitudes in Tibet.

Prevalence, associated risk factors, and phylogenetic analysis of Echinococcus granulosus isolated from freerange Tibetan pigs in Tibet, China.

The current study was performed to investigate the prevalence, associated risk factors exploration and phylogenetic analysis of Echinococcus granulosus (E. granulosus) genotypes isolated from Tibetan pigs. A total 373 Tibetan pigs were examined during 2014 and 2015, and the variables potentially associated with E. granulosus infection were explored with a multivariable logistic regression model. E. granulosus cysts (n=37) were collected from Tibetan pigs (lungs or livers). Fragments amplification of mitochondrial (mt) DNA of cox1 (shorter and longer) and atp6 were employed. The genotype of E. granulosus were identified by sequence and phylogenetic analysis. Results showed the prevalence of E. granulosus in Tibetan pigs was 9.9%. The prevalence of E. granulosus in male and female Tibetan pigs was 6.8% and 13.3%, with a significant difference in the two genders (P<0.05). In different seasons, the infection rate of E. granulosus in Tibetan pigs were ranged from 5.8% to 12.3%. E. granulosus infection rates in different growing stages of Tibetan pigs were ranged from 4.4% to 15.9%, with a statistical difference in the three stages (P<0.01). The prevalence of E. granulosus in Tibetan pigs were 7.9% in 2014 and 13.0% in 2015. Genders and growing stages were demonstrated to be risk factors to influence the prevalence significantly through multivariable logistic regression model. All the three fragments were successfully amplified from each of the 37 cysts. E. granulosus genotypes of G4 and G6 were identified by comparing with reference sequences of E. genotypes available at NCBI database and phylogenetic analysis by using MEGA software.

Seroprevalence of bluetongue virus infection in goats in the central China.

Seroprevalence of Bluetongue virus (BTV) in goats from Hubei was investigated by a commercial competitive enzyme-linked immunosorbent assay kits. Blood samples (n= 1157) were collected during the year 2014 and 2015. The results showed that 13.31% (CI 95% 11.4%-15.4%) serum samples were positive for BTV antibodies in goats in Hubei. The prevalence of BTV antibodies in each region ranged from 1.32% to 27.70%, and differences among the regions were statistically significant (p < 0.01). The prevalence of BTV in male and female goats was 14.23% (95% CI: 11.3, 17.6) and 12.58% (95% CI: 10.1, 15.4), respectively, no significant difference in genders (p > 0.05). In different seasons, the seroprevalence were 8.94% (95% CI: 5.6, 13.3) in spring; 18.31% (95% CI: 14.5, 22.7) in summer; 23.08% (95% CI: 17.0, 30.2) in autumn and 6.98% (95% CI: 4.6, 10.0) in winter, respectively with a significant difference of the prevalence in the different seasons (p < 0.01).

Molecular characterization of ascaris from Tibetan pigs by three mitochondrial markers of nad1, cox1 and cox2

Ascaris is a helminthic parasite, which infects a wide range of host species causing ascariasis, a predominant disease worldwide. This parasite causes significant economic losses to the pig industry. The current study was designed to determine the Ascaris nematode by the genetic characterization of three mitochondrial (mt) genes namely NADH dehydrogenase subunit 1 (nad1), cytochrome oxidase subunit 1 (cox1) and cytochrome oxidase subunit 2 (cox2). A high infection rate of Ascaris nematode has been found in Tibetan pigs at the slaughter houses in Tibet Autonomous Region of China. The nad1, cox1 and cox2 genes sequences collected from adult Ascaris individuals were amplified by polymerase chain reaction. The cloned-amplicons and the positive products were sequenced and phylogenetic analysis was performed. The results indicated that the Ascaris infecting the Tibetan pigs were Ascaris suum (A. suum). This is the first report on the isolation, identification and genetic characterization of three mitochondrial genomes (nad1, cox1, and cox2) of A. suum originated from Tibetan pigs at high altitudes in Tibet.

Molecular variability of five Cucumber mosaic virus isolates from China.

Cucumber mosaic virus (CMV) isolates are currently divided into two main groups, I and II according to their genomic sequences. The group I is further divided into two subgroups IA and IB. We performed a phylogenetic analysis of the genome regions containing 1a, 2a, 2b, coat protein (CP), and movement protein (MP) genes of 5 CMV isolates from China and other 28 CMV isolates available in the GenBank. The results indicated that CMV isolates could be genetically divided into three groups I, II, and III according to the genes encoding MP, CP, 1a, and 2a proteins and to the 2 groups according to the gene 2b. Group I could be further divided into two subgroups (IA and IB) according to the genes encoding CP, MP, 2a, and 2b proteins and to the three subgroups (IA, IB, and IC) according to the gene encoding 1a protein. Four of 5 examined Chinese CMV isolates belonged to the subgroup IB, while the remaining isolate was a natural inter-subgroup reassortant. We found that the 2b gene of CMV was under positive selection, while the other genes were under negative selection. No evidence of the selection associated with a host adaptation or geographic distribution was found.

Value of two-dimensional echocardiography in selecting patients and balloon sizes for percutaneous balloon mitral valvuloplasty.

The effect of valvular and subvalvular morphologic features and balloon size/mitral anulus size ratio on results of valvuloplasty were prospectively studied in 38 consecutive patients undergoing mitral valvuloplasty. The severity of valvular and subvalvular disease was graded echocardiographically from grade I to IV (mild to severe) for immobility, thickening, calcification of mitral leaflets and subvalvular thickening and fusion, yielding a maximal total score of 16. The diastolic mitral anulus diameter was measured in the apical four chamber view. After valvuloplasty, the mitral valve area increased from 0.9 +/- 0.3 to 2.2 +/- 0.5 cm2 (p less than 0.001) with increasing mitral regurgitation in 12 (32%) of the 38 patients. Multiple stepwise analysis revealed that the ratio of balloon size and annular size and the severity of subvalvular disease are two independent factors that correlated significantly with the mitral valve area after valvuloplasty (multiple r = 0.65, p less than 0.0002). One of 34 patients with mild subvalvular disease of grade III or less had an unsatisfactory increase in mitral valve area to less than or equal to 1.5 cm2, whereas 3 of 4 patients with severe (grade IV) subvalvular disease had a valve area less than or equal to 1.5 cm2 (p less than 0.002) after valvuloplasty. The increase in mitral regurgitation after valvuloplasty correlated significantly with the ratio of balloon to mitral anulus size and the severity of subvalvular disease (multiple r = 0.53, p less than 0.003). (ABSTRACT TRUNCATED AT 250 WORDS)

Percutaneous mitral balloon dilatation by a new sequential single- and double-balloon technique.

Percutaneous balloon valvotomy by means of a new sequential single- and and double-balloon dilatation procedure was performed in 23 patients (aged 13 to 53 years) with severe rheumatic mitral stenosis. The dilatation procedure was initially performed with a small balloon to primarily dilate the stenotic valve for easier passage of a second balloon catheter and to make the procedure tolerable for severely ill patients; the procedure was then followed by two balloons to further increase the mitral valve area (MVA) for effective dilatation of the stenotic mitral orifice. The dilatation was successful in all patients; the mitral valve pressure gradient decreased from 19 +/ 6 to 5 +/ 2 mm Hg, the cardiac output increased from 4.0 +/ 0.5 to 5.2 +/ 0.6 L/min, and the MVA increased from 0.8 +/ 0.2 to 1.9 +/ 0.4 cm2 (p less than 0.01 each). The MVA after dilatation was relative to the effective balloon dilatation diameter selected (r = 0.57; p less 0.01). A small atrial septal defect was observed in 3 of 23 patients immediately after the dilatation procedure. Mild mitral regurgitation was produced in 3 of 23 patients by the dilatation. We conclude that the sequential single- and double-balloon dilatation procedure can effectively increase the MVA and improve hemodynamics in severe mitral stenosis and that the larger effective balloon diameter of 24.8 mm or more (12 +/ 18 mm of two balloons) is necessary for effective dilatation.