The complete mitochondrial genome of Gasterophilus intestinalis, the first representative of the family Gasterophilidae.

Gasterophilus spp. (Diptera: Gasterophilidae) has a worldwide distribution; however, no complete mitochondrial (mt) genome data is available for Diptera which has greatly impeded population genetics, phylogenetics, and systematics studies in Gasterophilidae. Mt genome is known to provide genetic markers for investigations in these areas, but complete mt genomic datasets have been lacking for many Gasterophilidae species. Herein, we present the complete mt genome of the third-stage larvae (L3) of Gasterophilus intestinalis from the stomach wall of naturally infected horses in Heilongjiang province (HLJ) and Xinjiang Uygur Autonomous Region (XJ), China. The complete mt genome of G. intestinalis was 15,687 bp (HLJ) and 15,660 bp (XJ) in length and consists of 37 genes, including 13 genes for proteins, 22 genes for tRNA, and 2 genes for rRNA. The gene arrangement is the same as those of Oestroidae species. Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes by Bayesian inference (BI) and maximum likelihood (ML), suggested that the families Gasterophilidae and Oestroidae were more closely related than to Tachinidae. The mt genome of G. intestinalis represents the first mt genome of any member of the family Gasterophilidae. These data provide novel mtDNA markers for studying the molecular epidemiology and population genetics of the G. intestinalis and its congeners.

Diagnostic efficacy of contrast-enhanced ultrasound, dynamic contrast-enhanced MRI combined with tumor markers AFP and DCP for primary hepatocellular carcinoma.

The purpose of this study was to investigate the diagnostic efficacy of contrast-enhanced ultrasound (CEUS) and dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) combined with tumor markers alpha-fetoprotein (AFP) and des-γ-carboxyl prothrombin (DCP) for primary hepatic carcinoma (PHC). A total of 70 patients with PHC (PHC group), 42 patients with liver cyst (benign liver disease group (BLDG)) and 30 healthy people (healthy group (HG)) were selected as the research objects. CEUS and DCE-MRI were performed by American GE Vivid E9 color Doppler ultrasound system and Siemens 1.5T magnetic resonance imager, respectively. The levels of AFP and DCP were detected by ABBOTT i2000SR chemiluminescence instrument and enzyme-linked immunoassay (ELISA), respectively. In DCE-MRI examination, the portal phase and prolonged phase were mostly low signal in T1-weighted imaging (T1WI) sequence, and arterial phase was mostly high signal in T2WI sequence. In CEUS, most lesions showed hyper-enhancement in arterial phase, and hypo-enhancement in portal phase and delayed phase. AFP and DCP levels in PHC group were significantly higher than that in BLDG group and HG group. There were statistically significant differences among the three groups. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of combined diagnosis were statistically significant when compared with CEUS, AFP and DCP alone and either AFP or DCP positive. CEUS, DCE-MRI combined with tumor markers AFP and DCP have high sensitivity, specificity and accuracy in the diagnosis of PHC, which can more accurately diagnose the lesion type, provide basis for further treatment, and is worthy of clinical application.

miR-211 inhibits proliferation, invasion and migration of cervical cancer via targeting SPARC.

Cervical cancer remains one of the most frequent gynecological malignancies among females around the world. Therefore, fully understanding the molecular mechanisms underlying the progression of cervical cancer may be critical for the development of effective therapeutic strategies against cervical cancer. The object was to evaluate the potential effect of miR-211 and verify its influence on the function of secreted protein acidic and rich in cysteine (SPARC) in cervical cancer. It was demonstrated that miR-211 was downregulated in cervical cancer cell lines (HeLa and C33A) and cervical cancer specimens, while SPARC expression level was higher in tumor tissues. We also revealed miR-211 upregulated expression could inhibit cells proliferation, migration and invasion in vivo. SPARC was confirmed as a direct and functional target of miR-211 and the inverse relationship between them was also observed. The results of the present study suggest that miR-211 reduced cancer growth, migration and invasion, and suppresses the SPARC expression in cervical cancer. This newly identified miR-211 may provide further insight into the progression and offers a promising target for cervical cancer therapy.

Sequence variation in three mitochondrial genes among Gasterophilus intestinalis isolates from two distinct regions in China.

Horse gastrointestinal myiasis caused by larvae of Gasterophilus spp. has a worldwide distribution. However, little information is available on the genetic variation of Gasterophilus spp. at the molecular level. In the present study, sequence variation was examined in three mitochondrial (mt) genes, namely mt cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunits 5 (nad5) and large subunit ribosomal RNA (rrnL), among G. intestinalis isolates originating from Heilongjiang Provience and Xinjiang Uygur Autonomous Region, China. The complete cox1, nad5, and rrnL gene sequences were amplified by PCR separately from adult G. intestinalis individuals and the amplicons were sequenced from both directions. The size of the sequences of cox1, nad5, and rrnL genes was 1539 bp, 1717 bp, and 1321 bp, respectively. The A + T contents of the sequences were 62.7-63.9% (cox1), 70-70.7% (nad5), and 74.5-74.9% (rrnL). The intra-specific sequence variations within G. intestinalis were 0.1-3.1% for cox1, 0.4-3.4% for nad5, and 0.1-1.5% for rrnL. Phylogenetic analyses based on the cox1 sequences using the Bayesian inference indicated that all the G. intestinalis isolates grouped together with high statistical support. These findings demonstrated clearly the usefulness of mt cox1, nad5, and rrnL sequences for the molecular identification and for studying population genetics of G. intestinalis in horses.

Genetic variability among Hymenolepis nana isolates from different geographical regions in China revealed by sequence analysis of three mitochondrial genes.

Hymenolepis nana is a common tapeworm that parasitizes in the small intestine of rodent animals and humans. The present study examined the sequence diversity of three mitochondrial (mt) genes namely NADH dehydrogenase subunits 5 (nad5), small subunit ribosomal RNA (rrnS), and ATPase subunit 6 (atp6) of H. nana from mice in different geographical regions of China. A part of the nad5 (pnad5), complete rrnS and atp6 genes were amplified separately from individual H. nana isolates using polymerase chain reaction (PCR) and then sequenced. The sequences of pnad5, rrnS, and atp6 were 710 bp, 704-711 bp, and 516 bp in length, respectively. The A + T contents of the sequences were 70.1-73.5% (pnad5), 70.1-71.7% (rrnS), and 76.6-77.9% (atp6). Sequence variation within H. nana was 0-1.4% for atp6, 0-1.7% for rrnS, and 0-0.7% for pnad5. The inter-specific sequence differences between H. nana and Hymenolepis diminuta were significantly higher, which was 31.6-31.7% (pnad5), 16.1-17.6% (rrnS), and 26.5-27.1% (atp6). Phylogenetic analysis based on the combined three sequences using the maximum parsimony (MP) method supported that H. nana is a species complex or "cryptic" species. These findings demonstrated clearly the usefulness of the three mtDNA sequences for population genetics and systematic studies of H. nana of human and animal health significance.

Complete Mitochondrial genome of an equine intestinal parasite, Triodontophorus brevicauda (Chromadorea: Strongylidae): the first characterization within the genus.

The complete mitochondrial (mt) genome sequence of Triodontophorus brevicauda, an intestinal equine nematode parasite was determined for the first time. The circular T. brevicauda mt genome is 14,305 bp in length and contains 36 genes, of which 12 code for protein, 22 for transfer RNA, and two for ribosomal RNA, and lacks atp8 mtDNA gene. Phylogenetic analysis based on the concatenated amino acid sequence of the 12 protein-coding genes was performed using three different tree-building methods. The Strongyloidea cluster divides into two large branches, and each nematode family included in our study forms an independent clade, though paraphyly confounds the issue at some nodes. T. brevicauda clusters together with Cylicocyclus insignis with high statistical support. The mtDNA data in this study not only provide a new mtDNA resource for phylogeny, but also become a novel and useful genetic marker for further studies on the identification, population genetics, and molecular epidemiology of the genus Triodontophorus in equine.