Argonaute 1 (AGO1) is a core component of the RNA-induced silencing complex (RISC) which plays a crucial role in small RNA-mediated gene silencing. AGO1 gene has been characterized in various plants, such as Arabidopsis and rice. However, there is no information about AGO1 in the medicinal plant species, Panax ginseng. Using the rapid amplification of cDNA ends technology (RACE), we cloned full-length PgAGO1 cDNA from Panax ginseng. It is 3 776 bp in length, including 204 bp of 5' UTR, 254 bp of 3' UTR, and 3 318 bp of ORF encoding 1106 amino acids. The molecular weight (MW) and theroretical isoelectric point (pI) of the deduced PgAGO1 protein is 122.22 kDa and 9.71, respectively. PgAGO1 shares 91.72% similarity with Arabidopsis AtAGO1 and contains three consered domains, including DUF1785, PAZ and Piwi, suggesting it is an authentic AGO. PgAGO1 was expressed in all of the tissues analyzed with the highest level in flowers and the lowest level in roots. The results provide useful information for further elucidating the function of AGO1 in Panax ginseng.
Amino Acid Sequence ; Argonaute Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Genes, Plant ; Molecular Sequence Data ; Panax ; genetics ; Plants, Medicinal ; genetics ; Sequence Alignment

OBJECTIVETo study the expressions of Piwil2 protein and mRNA in papillary thyroid carcinoma (PTC) and the relationship between Piwil2 and the invasion and metastasis of PTC.

METHODSImmunohistochemistry and in situ hybridization were used to detect the expression of Piwil2 protein and mRNA in 60 cases of PTC with the matched adjacent non-cancerous epithelium (NCE).

RESULTSThe positive rates of Piwil2 protein expression in PTC and NCE were 88.3% (53/60) and 10.0% (6/60) respectively, with significant difference (χ² = 73.654, P < 0.01). The positive rates of Piwil2 mRNA expression in PTC and NCE were 85.0% (51/60) and 6.7% (4/60) respectively, also with significant difference (χ(2) = 74.148, P < 0.01). Up-regulated expressions of Piwil2 protein and mRNA were related to the invasion and metastasis of PTC (P < 0.05).

CONCLUSIONSPiwil2 may play a role in the invasion and metastasis of PTC.

Adolescent ; Adult ; Aged ; Argonaute Proteins ; Carcinoma ; Carcinoma, Papillary ; Child ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Thyroid Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo analyze the amino acid sequence composition, secondary structure, the spatial conformation of its domain and other characteristics of Argonaute protein.

METHODSBioinformatics tools and the internet server were used. Firstly, the amino acid sequence composition features of the Argonaute protein were analyzed, and the phylogenetic tree was constructed. Secondly, Argonaute protein's distribution of secondary structure and its physicochemical properties were predicted. Lastly, the protein functional expression form of the domain group was established through the Phyre-based analysis on the spatial conformation of Argonaute protein domains.

RESULTS593 amino acids were encoded by Argonaute protein, the phylogenetic tree was constructed, and Argonaute protein's distribution of secondary structure and its physicochemical properties were obtained through analysis. In addition, the functional expression form which comprised the N-terminal PAZ domain and C-terminal Piwi domain for the Argonaute protein was obtained with Phyre.

CONCLUSIONSThe information relationship between the structure and function of the Argonaute protein can be initially established with bioinformatics tools and the internet server, and this provides the theoretical basis for further clarifying the function of Schistosoma Argonaute protein.

Animals ; Argonaute Proteins ; chemistry ; genetics ; Chemical Phenomena ; Cluster Analysis ; Computational Biology ; methods ; Models, Molecular ; Phylogeny ; Protein Conformation ; Schistosoma ; chemistry ; genetics ; Sequence Homology, Amino Acid

In about half the cases of involuntary childlessness, a male infertility factor is involved. The PIWI-LIKE genes, a subclade of the Argonaute protein family, are involved in RNA silencing and transposon control in the germline. Knockout of murine Piwi-like 1 and 2 homologs results in complete infertility in males. The aim of this study was to analyze whether the mRNA expression of human PIWI-LIKE 1-4 genes is altered in ejaculated spermatozoa of men with impaired sperm characteristics. Ninety male participants were included in the study, among which 47 were with normozoospermia, 36 with impaired semen characteristics according to the World Health Organization (WHO) manual, 5^th edition, and 7 with azoospermia serving as negative control for the PIWI-LIKE 1-4 mRNA expression in somatic cells in the ejaculate. PIWI-LIKE 1-4 mRNA expression in the ejaculated spermatozoa of the participants was measured by quantitative real-time PCR. In nonazoospermic men, PIWI-LIKE 1-4 mRNA was measurable in ejaculated spermatozoa in different proportions. PIWI-LIKE 1 (100.0%) and PIWI-LIKE 2 (49.4%) were more frequently expressed than PIWI-LIKE 3 (9.6%) and PIWI-LIKE 4 (15.7%). Furthermore, a decreased PIWI-LIKE 2 mRNA expression showed a significant correlation with a decreased sperm count (P = 0.022) and an increased PIWI-LIKE 1 mRNA expression with a decreased progressive motility (P = 0.048). PIWI-LIKE 1 and PIWI-LIKE 2 mRNA expression exhibited a significant association with impaired sperm characteristics and may be a useful candidate for the evaluation of the impact of PIWI-LIKE 1-4 mRNA expression on male infertility.
Adult ; Argonaute Proteins/genetics* ; Azoospermia/genetics* ; Case-Control Studies ; Gene Expression ; Humans ; Infertility, Male/genetics* ; Male ; Middle Aged ; RNA, Messenger/metabolism* ; RNA-Binding Proteins ; Sperm Count ; Sperm Motility/genetics* ; Spermatozoa/metabolism* ; Young Adult

OBJECTIVETo observe the histological features of tumor-bearing tissues formed by human fibroblasts after reprograming by spermatogonial stem cell self-renewal key regulating gene Piwil2 (Piwil2-iCSC).

METHODSPiwil2-iCSC tumor spheroids-like colonies were selected for tumor formation assay in four nude mice. Pathological features of Piwil2-iCSC tumors were observed by histology. Stem cell markers and common triploblastic markers were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay and immunohistochemistry. Germ cell tumor markers were detected by immunohistochemical examination.

RESULTSTwo weeks after inoculation, subcutaneous tumors were formed in all the four nude mice with a tumor formation rate of 100%. In the Piwil2-iCSC tumor tissues, Piwil2-GFP(+) cells showed high-density nuclear expression and were widely observed in DAPI-stained sections. Numerous mitotic figure of the neoplastic cells were seen (>10 cells/field of vision under high magnification) in HE-stained sections. Enlarged abnormal cell nuclei were observed. RT-PCR assay showed that Piwil2-iCSC tumors still expressed Piwil2 and some self-renewal and pluripotent markers of stem cells and some markers of triploblastic differentiation. Immunohistochemical staining showed that the tumors expressed stem cell markers, triploblastic markers and germ cell tumor markers AFP and HCG.

CONCLUSIONSPiwil2-iCSC tumors are probably undifferentiated embryonic small cell carcinoma, most likely to be immature teratoma, mixed with yolk sac tumor and choriocarcinoma components. It can be used as a useful model for the research of origin or genesis mechanism of cancer stem cells and the treatment of relevant tumors.

Adult Stem Cells ; Animals ; Argonaute Proteins ; genetics ; Cellular Reprogramming Techniques ; Choriocarcinoma ; pathology ; Endodermal Sinus Tumor ; pathology ; Fibroblasts ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Mice ; Mice, Nude ; Neoplasms, Germ Cell and Embryonal ; chemistry ; genetics ; pathology ; Neoplastic Stem Cells ; chemistry ; pathology ; Real-Time Polymerase Chain Reaction ; Spheroids, Cellular ; Teratoma ; pathology ; Time Factors