Circular RNAs (circRNAs) are endogenous non-coding RNAs with covalently closed structures, which have important functions in plants. However, their biogenesis, degradation, and function upon treatment with gibberellins (GAs) and auxins (1-naphthaleneacetic acid, NAA) remain unknown. Here, we systematically identified and characterized the expression patterns, evolutionary conservation, genomic features, and internal structures of circRNAs using RNase R-treated libraries from moso bamboo (Phyllostachys edulis) seedlings. Moreover, we investigated the biogenesis of circRNAs dependent on both cis- and trans-regulation. We explored the function of circRNAs, including their roles in regulating microRNA (miRNA)-related genes and modulating the alternative splicing of their linear counterparts. Importantly, we developed a customized degradome sequencing approach to detect miRNA-mediated cleavage of circRNAs. Finally, we presented a comprehensive view of the participation of circRNAs in the regulation of hormone metabolism upon treatment of bamboo seedlings with GA and NAA. Collectively, our study provides insights into the biogenesis, function, and miRNA-mediated degradation of circRNAs in moso bamboo.
RNA, Circular/metabolism* ; Multiomics ; Poaceae/metabolism* ; Seedlings/genetics* ; Hormones/metabolism* ; MicroRNAs/metabolism* ; Gene Expression Regulation, Plant

Objective To assess the drug sensitivity of pancreatic cancer cells based on real-time cell analysis and provide a reference for individualized diagnosis and treatment of pancreatic cancer.Methods Three human pancreatic cancer cells lines SW1900, Capan-2 and PANC-1 were selected and treated with gemcitabine hydrochloride and tegafur gimeracil oteracil potassium capsules, respectively.After 24 hours of culture, the cells were treated with the two drugs in gradient concentration.The cell growth curves before and after the drug administration was monitored using a real-time cells analyzer and the growth inhibition rates (IC50) of the drugs of the pancreatic cancer cells were calculated.At the same time, the cells in the cell culture plate were treated with the drug, and acridine orange/ethidium bromide (AO/EB) staining and laser scanning confocal microscopy were performed to observe the changes of cells after the drug administration.Results 72 hours after the drug administration, IC50 values for the three cell lines were different.The IC50 values of gemcitabine hydrochloride for SW1900, Capan-2 and PANC-1 cells were 1.69 μmol/L, 10.05 μmol/L and 12.74 μmol/L, respectively.The IC50 values of tegafur capsule for SW1900, Capan-2 and PANC-1 cells were 180.29 μmol/L, 765.70 μmol/L and 95.57μmol/L, respectively.AO/EB staining confirmed the reliability of IC50.Conclusions SW1900 and Capan-2 cells can be used as the control for gemcitabine hydrochloride and tegafur gimeracil oteracil potassium capsules to establish cell models for drug screening in vitro, which provides a reference for the application of the technology in anticancer drugs screening.

Objective To develop a more convenient and stable method for assessment of drug sensitivity of prostate cancer based on real-time cell analysis system as a reference for clinical treatment.Methods Human prostate cancer VCaP, DU145, PC-3, PC-3M-2B4 and PC-3M-IE8 cells were chosen to detect the sensitivity to three drugs, docetaxel, cabazitaxel and abiraterone acetate.Serial dilutions of the three drugs were used to treat the cell culture for 24 hours.The drug-induced effects on the cell lines after an incubation of 24 hours were recorded by the real-time cell analysis system to determine the half maximal inhibitory concentration (IC50).Results Docetaxel showd IC50 of 8.81 nmol/L, 11.61 nmol/L, 1.78 nmol/L, 1.44 nmol/L, 8.69 nmol/L for VCaP, DU145, PC-3, PC-3M-2B4, PC-3M-IE8 cells, respectively.Cabazitaxel showed IC50 of 3.73 nmol/L, 3.96 nmol/L, 10.41 nmol/L, 5.43 nmol/L, and 7.37 nmol/L, respectively, for the five cell lines.Abiraterone acetate showed IC50 of 8.34 μmol/L, 8.60 μmol/L, 24.20 μmol/L, 8.59 μmol/L, and 13.21 μmol/L for the five cell lines.Conclusions PC-3M-2B4 and DU145, VCaP and PC-3 cells can be used as control for docetaxel, cabazitaxel and abiraterone acetate to establish cell models for the drug screening in vitro and to provide reference for clinical applications.

Objective To explore the effect of tert?butylhydroquinone(tBHQ)on ultraviolet B(UVB)?induced oxidative damages in human im?mortalized keratinocytes(HaCaT),and discuss its mechanism. Methods The cultured HaCaT cells were randomly divided into 4 groups:control group(G1),ultraviolet irradiation group(G2),25μmol/L tBHQ pretreatment before ultraviolet irradiation group(G3),and 50μmol/L tBHQ pre?treatment before ultraviolet irradiation group(G4). The content of reactive oxygen species was detected by DCFH?DA method,and the cell prolifera?tion was evaluated by MTT. Western blot was used to measure the protein expression of nuclear factor E2?related factor 2(Nrf2)in both nuclear fac?tions and whole?cell of HaCaT. The mRNA expressions of CAT and SRX were determined by real?time RT?PCR. Results The content of reactive oxygen species in HaCaT cells was increased,and the cell proliferation rate was decreased significantly after ultraviolet irradiation. The pretreatment of 25 and 50μmol/L tBHQ can inhibit the UVB?induced oxidative damage in a dose?dependent manner in HaCaT cells. Compared with G2 group, tBHQ pretreatment could dose?dependently increase the level of Nrf2 protein in nuclear factions and whole?cell of HaCaT,and also the mRNA ex?pressions of CAT and SRX. Conclusion UVB irradiation can induce oxidative stress damages of HaCaT cells. tBHQ may inhibit the UVB?induced oxidative damages through enhancing Nrf2 expressions and nuclear translocation,then activating the transcription of the downstream antioxidant en?zymes CAT and SRX.

Objective To investigate the meridians and acupoints selection in the treatment of post-stroke spastic paralysis with Tuina (Chinese massage). Methods The literatures about Tuina therapy for post-stroke spastic paralysis were retrieved. The frequency of meridians and acupoints used was counted and analyzed. Results 99 papers were collected, which involved in 226 acupoints and 1602 times of application. The selected acupoints were distributed in all the fourteen meridians and 64.5% (1033/1602, 606 on upper limb, 427 on lower limb) of them were on the limbs. The acupoints of the Yang meridians was 75.0% (1200/1602) and the specific acupoints was 71.7% (1148/ 1602). Conclusion The acupoint selected was basically focused on the local areas in Tuina for post-stroke spastic paralysis, assisted with the involved meridians and distal acupoints. The acupoints of the Yang meridians were the first option, mainly on the extremities. The specific acupoints were the major components of the prescription, especially the Five-shu acupoints and the He acupoints.