Calcined oyster is a commonly used shellfish traditional Chinese medicine in clinical practice in China. During the processing of oysters, their microscopic characteristics are destroyed, and open-fire calcination can damage the DNA of oysters, making it difficult to identify the primary source. The establishment of a specific polymerase chain reaction(PCR) method for the identification of calcined oysters can provide a guarantee for the safety and clinical efficacy of the medicine and its processed products. With Ostrea gigas as an example, the DNA extraction method of decoction pieces and formula particles of calcined oysters was improved, and high-quality DNA was obtained. Based on the specific single nucleotide polymorphism(SNP) sites of O. gigas and the other two species, the specific identification primers were designed, and the site-specific identification method of formula granules of calcined oyster(O. gigas) was established. The specificity and applicability of the method were investigated. The results showed that when the annealing temperature was 54 ℃, and the cycle was 44 times, the PCR amplified products of calcined oyster(O. gigas) and its formula granules produced a single bright identification band at 102 bp, while the other two species of oysters, O. talienwhanensis Crosse and O. rivularis Gould, had no band. In this study, DNA extraction and PCR identification of animal medicinal materials by calcination were established for the first time, which provided a tool for solving the difficult identification of calcined decoction pieces and ensuring drug safety.
Animals ; Ostrea/classification* ; Polymerase Chain Reaction/methods* ; Polymorphism, Single Nucleotide ; DNA/genetics*

The N⁶-methylation of RNA adenosines (N⁶-methyladenosine, m⁶A) is an important regulator of gene expression with critical implications in vertebrate and insect development. However, the developmental significance of epitranscriptomes in lophotrochozoan organisms remains unknown. Using methylated RNA immunoprecipitation sequencing (MeRIP-seq), we generated transcriptome-wide m⁶A-RNA methylomes covering the entire development of the oyster from oocytes to juveniles. Oyster RNA classes display specific m⁶A signatures, with messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) exhibiting distinct profiles and being highly methylated compared to transposable element (TE) transcripts. Epitranscriptomes are dynamic and correspond to the chronological steps of development (cleavage, gastrulation, organogenesis, and metamorphosis), with minimal mRNA and lncRNA methylation at the morula stage followed by a global increase. mRNA m⁶A levels are correlated with transcript levels, and shifts in methylation profiles correspond to expression kinetics. Differentially methylated transcripts cluster according to embryo-larval stages and bear the corresponding developmental functions (cell division, signal transduction, morphogenesis, and cell differentiation). The m⁶A level of TE transcripts is also regulated and peaks during the gastrulation. We demonstrate that m⁶A-RNA methylomes are dynamic and associated with gene expression regulation during oyster development. The putative epitranscriptome implication in the cleavage, maternal-to-zygotic transition, and cell differentiation in a lophotrochozoan model brings new insights into the control and evolution of developmental processes.
Animals ; RNA/metabolism* ; Methylation ; RNA, Messenger/metabolism* ; Transcriptome ; Ostreidae/metabolism* ; RNA, Long Noncoding/metabolism*

Objective: To evaluate the application of real-time RT-PCR and semi-nested RT-PCR in the detection of norovirus in oysters and analyzing the genetic characteristics of the isolates. Methods: Real-time fluorescent RT-PCR and semi-nested RT-PCR were used to detect norovirus GⅠ/GⅡ in fresh oysters collected from the markets in Beijing from November 2014 to October 2015. The detection rate of the parallel test was also analyzed. In addition, the reliability of semi-nested RT-PCR was evaluated by agreement rate and consistency test (Kappa value). The positive products of norovirus GⅠ/GⅡ capsid protein region gene by semi-nested RT-PCR were sequenced. Software BioEdit 7.0.9.0 was used for sequence alignment, and software Mega 6.0 was used to construct the evolutionary tree. Results: In 72 samples, the detection rate of norovirus was 31.94% (23/72) by real-time RT-PCR, 38.89% (28/72) by semi-nested RT-PCR and 48.61% (35/72) by parallel test. The coincidence rate of the two methods was 73.61%, a moderate degree (Kappa value =0.43). A total of 13 norovirus strains were successfully sequenced, and 11 strains (7 GⅡ.17 strains, 2 GⅡ. 4 Sydney_ 2012 strains, 1 GⅡ. 1 strain and 1 GⅡ. 21 strain) were obtained from norovirus positive samples by two RT-PCR methods, two strains (1 GⅡ. 17 strain and 1 GⅡ. 3 strain) were obtained from real-time RT-PCR negative samples which were positive for norovirus by semi-nested RT-PCR. The similarity between these strains and reference strains from diarrhea patients, environmental sewage, and shellfish products were 84.4% - 100.0%. Conclusions: The parallel test of norovirus in oysters by two RT-PCR methods can improve the detection rate and detect more genotypes. Norovirus strains in oysters were highly homologous with reference strains from diarrheal patients, environmental sewage, and shellfish products. Therefore, surveillance, prevention and control for norovirus should be carried out in people who have frequent contacts with oysters and related environments.
Animals ; Beijing ; Humans ; Norovirus/genetics* ; Ostreidae ; RNA, Viral/genetics* ; Real-Time Polymerase Chain Reaction ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction

Aims: Bivalve molluscs are filter feeders that tend to accumulate and concentrate any suspended particles or bacteria from the water environment. Although they have been proven to accumulate heavy metals, investigation on the presence of antibiotic-resistant bacteria is still lacking. Therefore, this study examines the occurrence of antibiotic-resistant bacteria isolated from bivalve shellfish, namely, blood cockles (Anadara granosa) and green-lipped mussel (Perna viridis) purchased from markets in Johor, Malaysia. Methodology and results: Samples purchased were homogenized and then diluted. Viable cell count and bacterial isolation were performed using diluted samples followed by an antibiotic susceptibility test (ampicillin, ciprofloxacin and tetracycline) conducted on the pure isolates. The total viable count of bacterial colonies for cockle and mussel samples ranged from 1.1 × 107 to 4.4 × 109 CFU/mL and 1.2 × 109 to 4.3 × 109 CFU/mL. The numbers of colonies isolated from respective bivalves were 11 and 6. Generally, cockles isolates revealed higher resistance towards all three antibiotics at or above the Clinical and Laboratory Standard Institute (CLSI) threshold value. Meanwhile, mussel isolates showed full susceptibility to any ciprofloxacin concentration and tetracycline but exhibited resistance to ampicillin at a concentration exceeding the CLSI value. The number of drug resistance isolates in cockle and mussel samples decreased with increasing antibiotic concentration. The maximum number of antibiotics the mussel isolates were resistant to was two, whereas cockle isolates achieved three. Conclusion, significance and impact of study The outcome of this study concludes that some isolates from cockle and mussel samples can resist antibiotic concentration above the CLSI threshold value. Resistance of more than the CLSI threshold level revealed that these isolates could pose significant health risks especially when the bivalves are ingested raw or undercooked.
Drug Resistance, Bacterial ; Bivalvia

Animals ; Cadmium/toxicity* ; Dysbiosis/metabolism* ; Gastrointestinal Microbiome ; Glutamine/pharmacology* ; Methionine ; Mice ; Mice, Inbred C57BL ; Ostreidae ; Protein Hydrolysates ; Tyrosine

Crassostrea sikamea (C.sikamea) is an important edible and medicinal seafood in China. In the present study, a compound named flazin was separated and identified from the ethyl acetate extract of C.sikamea (EAECs) for the first time. In addition, the 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra zolium (MTS) assay revealed that EAECs and flazin inhibited the transformation of splenic lymphocytes in vitro. Moreover, flazin (20 μg·mL
Animals ; Carbolines ; Crassostrea ; Furans ; Lymphocytes ; Rats ; Rats, Sprague-Dawley ; Spleen

Aims: Escherichia coli O157:H7 is known to be transmitted via fecal-oral route, where water plays a role in the transmission process. Oysters as bivalves, bio accumulate pathogens from the water through filter feeding and are suspected to play a role as disease transmission vector. In Malaysia, the data on oyster’s microbiological quality are limited. Hence, it was vital to conduct oyster related studies in Malaysia. The main objectives of this study include the enumeration of most probable number (MPN) of fecal coliforms and E. coli and isolation of E. coli from oyster (Crassostrea iredalei) and water sample for the detection of 16S rRNA and HlyA (Hemolysin A) genes of E. coli O157:H7. Methodology and results: A total of 120 oysters and water samples (n=6) were collected from a fisherman village located in southern Malaysia. Total fecal coliforms and E. coli were determined using the MPN procedure. Colonies of E. coli were identified based on Gram staining, biochemical test, and PCR detection for the presence of 16S rRNA and HlyA gene of E. coli O157:H7. The enumeration results showed that the MPN of the fecal coliforms and E. coli found in the collected oyster samples do not meet the standard to be directed for human consumption (0.72 ± 0.19 × 104 MPN/100 g and 0.13 ± 0.03 × 10 4 MPN/100 g, respectively). The PCR assays showed that 16 out of the 104 (15.38%) of E. coli isolated from water and oysters showed the presence of HlyA gene. The phylogenetic tree analysis showed there were genetic relationships between the HlyA gene of the E. coli isolated in this study with the ones isolated from calf and human faeces. Conclusion, significance and impact of study The detection of Shiga toxin producing E. coli O157:H7 (HlyA gene) in cage cultured oysters (C. iredalei) and water from southern Malaysia was first time reported here. In the future, more study can be conducted to study the expression of the HlyA gene and confirm of its identity as E. coli O157:H7 using different target genes such as eaeA (encodes a 94 kD outer membrane protein called intimin) and Stx1 (Shiga toxin, Shigella dysenteriae type 1).
Escherichia coli O157 ; Crassostrea

Objective: Owing to antibiotic abuse and the subsequent development of antibiotic resistance, bacterial infection has become one of the most persistent unresolved problems. New antibacterial agents, especially those that are environmental-friendly, are urgently needed. Methods: Melanin extracted by filtration centrifugation and acid and proteolytic hydrolysis was characterized using UV, FTIR, TEM, and XPS. Photothermal conversion was calculated, and the bacteriostatic effects, and , were assessed by plate counting and ratios (%) of wound areas. Results: Natural melanin hydrolyzed by trypsin had good photothermal conversion effects, which resulted in superior bacteriostatic activities. The extracted melanin along with laser NIR irradiation at 808 nm promoted the healing of wounds infected by drug-resistant bacteria and was biocompatible according to toxicity tests and . Conclusion The present findings indicated a safe and efficient method of developing natural antibacterial agents.
Animal Shells ; chemistry ; Animals ; Anti-Bacterial Agents ; pharmacology ; Escherichia coli ; drug effects ; radiation effects ; Escherichia coli Infections ; drug therapy ; Melanins ; pharmacology ; Mytilus edulis ; chemistry ; Photochemical Processes ; Rats ; Rats, Sprague-Dawley ; Staphylococcal Infections ; drug therapy ; Staphylococcus aureus ; drug effects ; radiation effects ; Wound Healing

Intraosseous hemangioma is a rare, slow-growing, benign tumor of blood vessels. Primary hemangioma of the skull is a benign lesion that may appear as a palpable mass or accidentally detected during image evaluation. Simple radiography is the most commonly used technique to localize a lesion and computed tomography (CT) may help determine the effect of a lesion. We report a case of multifocal intraosseous calvarial hemangioma developed in the subgaleal plane of an elderly male patient. Ultrasonography examination revealed hyperechoic striated septae parallel to the skin and discontinuity of the focal cortex, however, the underlying bone cortex appeared relatively intact. No significant flow is observed on Doppler ultrasonography. Based on these evaluations, the mass was interpreted by a radiologist as a subgaleal lipoma. This case highlights the importance of additional CT examination in a patient presenting with a scalloping sign of the underlying calvarium. Clinicians also should be aware of the possibility of intraosseous calvarial hemangiomas in lesion. Furthermore, the proper choice of congenital vascular malformation term is still quite confusing with misconception present in the literature.
Aged ; Blood Vessels ; Hemangioma ; Humans ; Lipoma ; Male ; Pectinidae ; Radiography ; Skin ; Skull ; Ultrasonography ; Ultrasonography, Doppler ; Vascular Malformations

PURPOSE: This study examined the most efficient exercise position to activate the gluteus medius (GM) and tensor fascia latae (TFL) in hip abduction in side-lying (HA-SL), clam in side-lying (CL-SL), and sling bridging in side-lying (SB-SL), which are the most representative GM exercises. METHODS: Twenty-four healthy male adults aged from 20 to 40 years, whose body mass index was under 25, participated in this experiment. While all participants conducted three different positions with a counterbalanced manner, such as in AB, CL, and BR, activation of the GM and TFL was measured using 8-channel wireless EMG. Exercise was performed for 10 seconds three times in total with a five minute-break session. RESULTS: Significant differences in GM and TFL activation were observed among the three positions (p<0.05). The highest activation of 60.69 was observed in BR followed in order by 46.03 and 12.92 in HA-SL and CL, respectively. TFL activation in HA-SL was 42.01, followed in order by 35.98 and 14.01 in BR and CL, respectively. On the other hand, there was no significant difference in TFL muscle activation between BR and HA-SL. CONCLUSION: These findings suggest that both BR and HA-SL in GM can be done selectively. CL has remarkably low muscle activation in GM and TFL, which makes it less valuable in GM and TFL exercise. In conclusion, selective BR and HA-SL exercise should be applied to maximally and effectively activate the GM.
Adult ; Bivalvia ; Body Mass Index ; Exercise ; Fascia Lata ; Hand ; Hip ; Humans ; Male