ABSTRACT
The CRISPR/Cas9 gene editing technology directs Cas9 protein to recognize, bind and cleave the target site specifically by using artificial single-guide RNA (sgRNA), through non-homologous end joining or homologous end-recombinant repair mechanisms of cells, which can be engineered to knockout or knock-in of genomes. RIG-I is a pattern recognition receptor that recognizes the 5'-triphosphate-containing RNA in the cytoplasm and activates IRF3/7 and NF-κB by interacting with the downstream signaling molecule MAVS, thus initiating the expression of type I interferons and inflammatory factors. Previous studies found that influenza B virus (IBV) can up-regulate the expression of RIG-I. In the present study, to explore whether RIG-I is the major receptor for IBV to active the antiviral innate immune response and its effect on IBV replication, RIG-I gene in 293T cells was knocked out by CRISPR-Cas9 system, and a stable RIG-I knockout 293T (RIG-I(-/-) 293T) cell line was screened by puromycin pressure. The results of Western blotting showed that RIG-I was not expressed in this cell line after IBV or Sendai virus (SeV) infection, indicating that the RIG-I(-/-) 293T cell line was successfully constructed. The transcription levels of interferons, inflammatory factors and interferon-stimulated genes in RIG-I(-/-) 293T cells which were infected by IBV decreased significantly compared with those in wild-type 293T cells. Moreover, the phosphorylation of p65 and IRF3 were not detected in IBV or SeV infected RIG-I(-/-) 293T cells. It is indicated that the expression of cytokines mainly depends on the RIG-I-mediated signaling pathway at the early stage of IBV infection. Furthermore, the multi-step growth curves of IBV in the wild type and RIG-I(-/-) 293T cells showed that RIG-I inhibited the replication of IBV. Collectively, the RIG-I knockout 293T cell line was successfully constructed. We found that RIG-I is the main receptor for IBV to active the antiviral innate immune response and is critical for inhibiting IBV replication, which lays the foundation for further study of IBV infection mechanism.
ABSTRACT
Objective To explore the correlation between the detection rate of diffusion weighted imaging(DWI) in diagnosis prostate cancer and the Gleason score.Methods Ninety-one cases of intact pathological data and clinically suspected prostate cancer were collected.All cases were undertaken the routine sequence (TIWI/T2WI) and DWI (value b=600 s/mm2) examination.The cases of pathologically diagnosed prostate cancer were performed the Gleason score.Results Among 91 cases,44 cases were benign prostatic hyperplasia(BPH) and 47 cases were prostate carcinoma(PCa).The ADC values in the BPH group and PCa group were (1.250±0.263)× 10-3mm2/s and (0.955±0.191)× 10-3mm2/s.According to Gleason score,the PCa group was re-divided into three sub-groups:high differentiation,middle differentiation and low differentiation,the apparent diffusion coefficient(ADC) values were (1.158±0.173)× 10-3mm2/s,(1.050±0.192)× 10-3mm2/s and (0.891±0.241) × 10-3 mm2/s respectively.Their diagnostic accuracy rates for diagnosing PCa were 63.6％,78.3％ and 84.6％ respectively.The ADC value of different pathological groups in PCa with Gleason score is negatively correlated with the Gleason score.The accuracy rate of DWI in diagnosis of PEa is positively correlated with the Gleason score.Conclusion The lower prostate cancer differentiation,the higher Gleason score and the higher accuracy of DWI in diagnosing PCa.
ABSTRACT
Objective Based on the diagnostic technique of electric meridian detection, to observe the clinical efficacy of acupuncture in treating angioneurotic headache.Method Totally 106 patients with angioneurotic headache were randomized into a treatment group and a control group, 53 cases in each group. The treatment group received acupuncture based on the electric meridian detection, while the control group received medication treatment. The hemorheology indexes [plasma viscosity (PV), erythrocyte sedimentation rate (ESR)] and McGill Pain Questionnaire (MPQ) [Pain Rating Index (PRI), Visual Analogue Scale (VAS), and Present Pain Intensity (PPI)] were observed before and after intervention, and the clinical efficacies were compared. Result The total effective rate was 88.7% in the treatment group versus 71.7% in the control group, and the between-group difference was statistically significant (P<0.05). After intervention, the hemorheology indexes and MPQ scores were significantly changed in both groups (P<0.01,P<0.05).Conclusion Acupuncture based on electric meridian detection is an effective method in treating angioneurotic headache.