CircRRAS2 promotes myogenic differentiation of bovine MuSCs and is a novel regulatory molecule of muscle development.

The proliferation and myogenic differentiation of muscle stem cells (MuSCs) are important factors affecting muscle development and beef quality. There is increasing evidence that circRNAs can regulate myogenesis. We found a novel circRNA, named circRRAS2 that is significantly upregulated in the differentiation phase of bovine MuSCs. Here, we aimed to determine its roles in the proliferation and myogenic differentiation of these cells. The results showed that circRRAS2 was expressed in several bovine tissues. CircRRAS2 inhibited MuSCs proliferation and promoted myoblast differentiation. In addition, chromatin isolation by using RNA purification and mass spectrometry in differentiated muscle cells identified 52 RNA-binding proteins that could potentially bind to circRRAS2, in order to regulate their differentiation. The results suggest that circRRAS2 could be a specific regulator of myogenesis in bovine muscle.HighlightsCircRRAS2 expression is higher in DM cells than in GM cells.CircRRAS2 could significantly inhibit the proliferation and apoptosis of bovine MuSCs.CircRRAS2 promotes the differentiation of bovine MuSCs into myotubes.CircRRAS2 may exert regulatory effects through multiple RNA binding proteins.

CircUBE2Q2 promotes differentiation of cattle muscle stem cells and is a potential regulatory molecule of skeletal muscle development.

BACKGROUND: The growth and development of muscle stem cells (MuSCs) are significant events known to affect muscle plasticity, disease, meat production, and meat quality, which involves the types and functions of mRNA and non-coding RNA. Here, MuSCs were cultured from Guangxi fetal cattle. RNA sequencing was used to analyze the RNA expression of mRNA and non-coding RNAs during the cell proliferation and differentiation phases. RESULTS: Two thousand one hundred forty-eight mRNAs and 888 non-coding RNAs were differentially expressed between cell proliferation and differentiation phases, including 113 miRNAs, 662 lncRNAs, and 113 circRNAs. RT-qPCR verified the differential expression levels of mRNAs and non-coding RNAs, and the differentially expressed circUBE2Q2 was subsequently characterized. Expression profile analysis revealed that circUBE2Q2 was abundant in muscle tissues and intramuscular fat. The expression of cricUBE2Q2 was also significantly upregulated during MuSCs myogenic differentiation and SVFs adipogenic differentiation and decreased with age in cattle muscle tissue. Finally, the molecular mechanism of circUBE2Q2 regulating MuSCs function that affects skeletal muscle development was investigated. The results showed that circUBE2Q2 could serve as a sponge for miR-133a, significantly promoting differentiation and apoptosis of cultured MuSCs, and inhibiting proliferation of MuSCs. CONCLUSIONS: CircUBE2Q2 is associated with muscle growth and development and induces MuSCs myogenic differentiation through sponging miR-133a. This study will provide new clues for the mechanisms by which mRNAs and non-coding RNAs regulate skeletal muscle growth and development, affecting muscle quality and diseases.

[Differentially expressed proteins in serum among different Chinese medical syndrome types of primary liver cancer in the perioperative period of interventional treatment].

OBJECTIVE: To explore different expressions of serum proteins among different Chinese medical syndrome types of primary liver cancer (PLC) in the perioperative period of interventional treatment, and to explore its significance. METHODS: Totally 154 PLC patients were assigned to Gan depression syndrome (GDS, 37 cases), Pi deficiency syndrome (PDS, 45 cases), dampness heat syndrome (DHS, 18 cases), blood stasis syndrome (BSS, 28 cases), and yin deficiency syndrome (YDS, 26 cases). The mass spectra of serum proteins were analyzed by using surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). By combining the features of Chinese medical syndromes, the different expressions of serum proteins among different Chinese medical syndrome types of PLC in the perioperative period of interventional treatment were explored. RESULTS: One week before interventional treatment, there was statistical difference in the expression of serum protein peak with mass-to-charge ratio (M/Z) being 3,392, 4,970, 5,911, 6,200, and 8,575 Da (P <0.05, P < 0.01). The aforesaid differentially expressed protein peaks occurred simultaneously in PDS and BSS. One week after interventional treatment, the expression of the serum protein peak was down-regulated in YDS syndrome with M/Z being 8,575 Da, showing statistical difference (P <0.01). CONCLUSION: Different peaks of serum proteins occurred in different Chinese medical syndrome types of PLC in the perioperative period of interventional treatment.

[Observation of serum protein fingerprinting in primary liver cancer patients of different Chinese medical syndromes before and after interventional treatment].

OBJECTIVE: To explore changes of serum protein fingerprinting in primary liver cancer (PLC) patients of different Chinese medical syndromes before and after interventional treatment detected by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). METHODS: Totally 154 PLC patients were assigned to 5 groups, i.e., Gan depression syndrome (GDS, 37 cases), Pi deficiency syndrome (PDS, 45 cases), dampness heat syndrome (DHS, 18 cases), blood stasis syndrome (BSS, 28 cases), yin deficiency syndrome (YDS, 26 cases). The mass spectra of serum proteins was analyzed by using SELDI-TOF-MS. Then the correlation between Chinese medical syndrome types and the mass spectra of serum proteins was explored before and after interventional treatment. RESULTS: Compared with the healthy control group, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in DHS with M/Z being 5 710 Da, in YDS with M/ Z being 6 992 Da, while it was up-regulated in PDS with M/Z being 5 816 Da and in BSS with M/Z being 4 297 Da, showing statistical difference (P < 0.01). Compared with before intervention, the expression of serum proteins peak was down-regulated in GDS with M/Z being 6 589 and 4 182 Da, in PDS with M/Z being 5 816 Da, in DHS with M/Z being 5 710 Da in BSS with M/Z being 4 297 Da, while it was up-regulated in YDS with M/Z being 6 992 Da, showing statistical difference (P < 0.05, P < 0.01). CONCLUSION: There was statistical difference in changes of serum protein fingerprinting in PLC patients of different Chinese medical syndromes before and after interventional treatment.

E2F1-induced long non-coding RNA MCF2L-AS1 modulates Cyclin D1 mRNA stability through ELAVL1 to induce Gefitinib resistance in non-small cell lung cancer.

PURPOSE: Gefitinib is a widely used therapeutic drug for non-small cell lung cancer (NSCLC), and its acquired resistance has become one of the barriers to the successful use of the drugs to treat NSCLC patients. Long non-coding RNA (lncRNA) has an essential role in developing cancer drug resistance. Hence, this study aimed to investigate the effect and modulatory mechanisms of lncRNA MCF2L-AS1 in Gefitinib resistance in NSCLC. METHODS: IBEAS-2B and A549 cells and human NSCLC tissues were used. A549/GR cell line was constructed by continuous exposure to Gefitinib. Cell viability, apoptosis, migration, colony formation, and protein expression studies were done in transfected cells. Interactions of MCF2L-AS1, ELAVL1, and Cyclin D1 (CCND1 was also investigated. RESULTS: In patients with Gefitinib-resistant NSCLC, MCF2L-AS1 and CCND1 were both up-regulated. Knockdown of MCF2L-AS1 reduced Gefitinib-resistant NSCLC cell progression, indicating that inhibition of MCF2L-AS1 restrained Gefitinib-resistant NSCLC. Mechanically, MCF2L-AS1 enhanced CCND1 mRNA stability via combining with ELAVL1, thereby elevating the resistance of NSCLC cells to Gefitinib. Moreover, E2F1 could transcriptionally up-regulate MCF2L-AS1. CONCLUSION: The results manifest that lncRNA MCF2L-AS1, as an oncogene of NSCLC, controls CCDN1 via ELAVL1 to drive the growth of NSCLC cells and Gefitinib resistance.

Combined effects of low-dose gambogic acid and NaI131 in drug-resistant non-small cell lung cancer cells.

Radioactive seed brachytherapy is a method for treating drug-resistant, late-stage non-small cell lung cancer (NSCLC). To elucidate the mechanism of low-dose gambogic acid (GA) and NaI131 in drug-resistant NSCLC cells, the human NSCLC A549 cell line and the drug-resistant A549/cisplatin (DDP) and A549/Taxol cell lines were treated with NaI131, low-dose GA or a combination of both in the present study; the control group of each cell line was treated with phosphate-buffered saline (PBS). Following treatment, cell proliferation, apoptosis and cell cycle analysis was performed. Apoptosis-related proteins, namely CDK1, cyclin B, mutant p53 (mtp53), heat shock protein 90 (HSP90), Bax and Bcl-2, and P-glycoprotein 1 (P-gp), which is known to confer resistance to chemotherapy, were detected using western blotting and immunofluorescence analysis. mRNA levels of p53 and HSP90 were measured using reverse transcription-quantitative PCR. Compared with the PBS control group, the A549, A549/DDP and A549/Taxol cells treated with NaI131, GA or a combination of the drugs exhibited G2/M arrest and increased percentages of total apoptotic cells, as well as significantly decreased protein levels of CDK1, cyclin B, mtp53, HSP90, Bcl-2 and P-gp, increased protein levels of Bax and decreased mRNA levels of p53 and HSP90. The changes in the combination group were the most evident and were significantly different from the other groups (P<0.001). In conclusion, low-dose GA may be a potential radionuclide sensitizer.

Two new species of Araneus Clerck, 1757 (Araneae, Araneidae) and first description of A. wulongensis male from China.

Two new species of Araneus Clerck, 1757 are described: A. conexus sp. nov. (♂♀) and A. digitatus sp. nov. (♂♀) from Yunnan and Hubei provinces. The male of A. wulongensis Song & Zhu, 1992 is described here for the first time. All species treated in this study belong to A. strurmi species group. Detailed description and illustrations of somatic features, and copulatory organs as well as distribution maps are provided.

Two new species of Yaginumaella, Prószynski 1976 from Yunnan, China (Araneae, Salticidae).

The present paper deals with two new species of Yaginumaella, Yaginumaella lushuiensissp. n. (female and male) and Yaginumaella pseudoflexasp. n. (female and male). The male of Yaginumaella lushuiensissp. n. differs from the related species Yaginumaella flexa Song & Chai, 1992 in ventral view of palpal organ. The female of Yaginumaella lushuiensissp. n. differs from the related species Yaginumaella urbanii Zabka, 1981 by: 1) hoods locate at the anterior area of epigynum and far away from the copulatory openings; 2) epigynum about circular; 3) copulatory openings transverse. The male of Yaginumaella pseudoflexasp. n. differs from the related species Yaginumaella bulbosa Peng, Tang & Li, 2008 in ventral view of palpal organ: 1) basal portion of embolus touches the margin of genital bulb. 2) distal portion of tibial apophysis covers the posterior margin of cymbium and far away from the margin of genital bulb. The female of Yaginumaella pseudoflexasp. n. differs from the related species Yaginumaella urbanii Zabka, 1981 by: epigynum about as long as wide; hoods locate at the anterior area of the epigynum, above the outside area of the copulatory openings and far away from the copulatory openings. Photos of body and copulatory organs, line drawings of copulatory organs, as well as the locality map are provided. Descriptions of morphology are given.