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Objective: To investigate the infection rate and genotype distribution of Enterocytoon (E.) bieneusi in farmed black goats from the Hainan Province, China. Methods: A total of 341 fresh fecal samples were collected from black goats farmed in five different locations of the Hainan Province, China. E. bieneusi was examined and genotyped through PCR and sequencing analysis of the internal transcribed spacer (ITS) region of this pathogen. Results: The average prevalence of E. bieneusi in black goats from from the five locations was 24.0% (82/341) ranging from 6.3% (4/63) to 37.2% (32/86) (χ2 =17.252, P<0.01). The detected 82 E. bieneusi isolates belonged to eight ITS genotypes including six known genotypes (AHG1, CHG2, CHG3, CHG5, CM21 and D) and two novel genotypes (HNG-Iand HNG-II). Amongst the genotypes, CHG5 was the the most prevalent with a prevalence of 57.3% (47/82), followed by CHG3 (28.0%, 23/82), CHG2 (4.9%, 4/82), CM21 (3.7%, 3/82), D (2.4%, 2/82), AHG1 (1.2%, 1/82), HNG-I(1.2%, 1/82) and HNG-II(1.2%, 1/82). In those genotypes, only genotype D was found in humans previously. Conclusions: This represents the first report identifying E. bieneusi in black goats from Hainan Province of China. The results indicate that E. bieneusi has a high prevalence and a wide distribution in those animals from Hainan Province, but the risk of zoonotic transmission of E. bieneusi from them to human is low.
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Objective:To conduct in-depth study of the distribution and diversity of viruses in poultry is of great importance in monitoring the emergence of interspecies transmission of novel viruses that may cause epidemics with public health significance. Poultry is an economically important source of meat, eggs and feathers which plays an important role as natural reservoirs of many pathogenic viruses. Compared with wild animals, poultry have more frequent interactions and therefore opportunities to transmit their viruses to human.Methods:To study the viromes of different types of poultry in Hainan, China, we used metagenomics for deep viral nucleic acid sequencing of the faecal samples collected from chickens, ducks and pigeons from a live poultry market in Haikou.Result:The poultry viromes were identified by sequence similarity comparisons of viral reads (BLASTxE score, <5) against viral reference database. A total of 15 309 viral reads were obtained, approximately 13 063, 1 370 and 876 viral reads were generated from the chicken, duck, pigeon faecal samples, respectively. The majority of the sequences were homologous to the animal virus of Adenoviridae, Herpeaviridae, Picobirnaviridae, Reoviridae, Retroviridae, Circoviridae, Paramyxoviridae, Astroviridae, Caliciviridae, Coronaviridae, Picornaviridae, and Orthomyxoviridae. The VP4 and VP7 segments of a pigeon rotavirus, similar to fox rotavirus in group A, were sequenced and phylogenetically analyzed. The near full genome of a pigeon circovirus was also analyzed.Conclusion:The major types of poultry in a Haikou harbor many different families of viruses in their feces which may have the potential for interspecies transmissions. Further studies should be conducted to identify the most prevalent and important viruses among a larger number of poultry in Haikou and other areas in Hainan.
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Objective:Torque teno virus(TTV), are reported in a wide range of mammals. In this study, we sequenced and analyzed the complete genome of a genetic variant of Rodent TTV, RoTTV3-HMU1 (Hainan Medical University). The virus was harbored by a Rattus norvegicus in the residential areas of Hainan Island, China.Methods:Torque Teno virus (TTV) was found widely distributed throughout the world infecting an extensively wide range of mammals .We extracted the viral DNA from a Rattus norvegicus liver which was caught from the residential areas of Hainan Island. Purifying the amplicons in the range of 250-500 bp. Then Five hundred nanograms DNA was subjected to high-throughput sequencing. The contigs were compared with the NCBI nucleiotide database, designed the primers to cover the genome by PCR amplification and amplicons of each PCR which have been cloned and sequenced. Finally the genome was annotated by using NCBI ORF finder and FGENESV0. Phylogenetic analysis was implemented by the neighbor-joining method in the MEGA6 software package.Results:We sequenced the complete genome of a genetic variant of Rodent TTV, RoTTV3- HMU1. The genomic sequence of RoTTV3-HMU1 has been deposited in GenBank under accession number MF688246.1. The complete genome of RoTTV3-HMU1 is 2 570 nucleotides (nt) in length with a G+C content of 46.93%. RoTTV3-HMU1 encoded 3 unidirectional overlapping open reading frames (ORF). Sequence analysis indicated that the genome of RoTTV3-HMU1 virus was most closely related to RN_2_15 (GenBank accession no. KM668486.1). Phylogenetic analysis based on both ORF1 and the total genome sequence placed RoTTV3-HMU1 in to the clad RoTTV3 of the RoTTV.Conclusions:Hainan Island faces mainland across the sea, however, the same genotype of RoTTV was identified in both Hainan Island and the other part of China. The detection of RoTTV3-HMU1 contributed to a better understanding about the origin and evolution of RoTTV.
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Objective:Microsporidia have been rapidly emerging as pathogens in both immunocompromised and immunocompetent humans. Enterocytozoon bieneusi (E. bieneusi) is the most common microsporidial species found in human. E. bieneusi has also been found in a wide range of animals and is considered to be a potentially important zoonotic pathogen. The epidemiological and genetic characterization of E. bieneusi among long-tailed macaques [Macaca fascicularis (M. fascicularis) is not fully understood. Here, we conducted the first molecular epidemiological investigation of E. bieneusi among M. fascicularis in Hainan Province, the southernmost part of China.Methods:A total of 193 fecal specimens of M. fascicularis were collected from a breeding base housing non-human primates for experimental use in Hainan Province, China. E. bieneusi was identified and genotyped by nested PCR analysis of the internal transcribed spacer (ITS) region of the rRNA gene. Phylogenetic analysis was performed by constructing a neighboring-joining tree of the ITS gene sequences using MEGA6.Results:A total of 59 (30.6%) of the M. fascicularis were PCR-positive for E. bieneusi. All 59 samples were sequenced successfully and 16 ITS genotypes were identified. These included nine known genotypes: Type IV (n=19), D (n=11), CM1 (n=8), PigEBITS7 (n=4), Pongo2 (n=4), Peru 8 (n=3), Peru11 (n=1), WL21 (n=1) and CM2 (n=1). Additionally, seven novel genotypes named as HNM-I to HNM-VII (one each) were identified. Importantly, genotypes D, Type IV, Peru8, PigEBITS7, and Peru11, which were the predominant (38/59, 64.4%) genotypes identified among M. fascicularis in this study, are also well-known human-pathogenic genotypes. All the genotypes of E. bieneusi identified in this study, including the seven novel ones, belonged to zoonotic group 1.Conclusions:This is the first report of the identification of E. bieneusi in M. fascicularis in Hainan Province, China. The findings of numerous known human-pathogenic types and seven novel genotypes (HNM-I to HNM-VII) of E. bieneusi all belong to zoonotic group 1 indicate the possibility of transmission of this important pathogenic parasite between M. fascicularis and humans.
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<p><b>OBJECTIVE</b>To investigate the effects of metformin on proliferation and apoptosis in multiple myeloma cell line RPMI8226 and U266, and to clarify the molecular mechanism of proliferation inhibition and apoptosis induced by metformin.</p><p><b>METHODS</b>RPMI8226, U266 cells were treated with 0, 5, 10, 20, 40, 80 mmol/L of metformin for 24, 48 and 72 hours, then the inhibition rate was detected by CCK-8; RPMI8226 cells were treated with 0, 10, 20, 40 mmol/L of metformin for 48 hours, the apoptosis rates were detected by flow cytometry with Annexin-V-FITC/PI double staining; RPMI8226 cells were treated with 0, 5, 10, 20 mmol/L of metformin for 48 hours, the expressions of Caspase-3, PARP, STAT3, p-STAT3, BCL-2, Cyclin D1 and P21 were detected by Western blot.</p><p><b>RESULTS</b>The inhibition rate increased in RPMI8226 and U266 cells treated with metformin in the dose- (r=0.982, r=0.967, P<0.05) and time-dependent (r=0.956, r=0.962, P<0.05) manner; the apoptosis rate increased(r=0.976, P<0.05) in RPMI8226 cells treated with metformin; it also was found that procaspase-3 was degraded and PARP was cleaved when treated with metformin. Proliferation inhibition and apoptosis of RPMI8226 cells were related with inhibition of STAT3 phosphorylation, down-regulation of BCL-2 and Cyclin D1, and up-regulation of P21.</p><p><b>CONCLUSION</b>Metformin can inhibit the proliferation and induce apoptosis of RPMI8226 and U266 cell lines, which may be related to down-regulation of STAT3 signal transduction pathway.</p>