Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there is still a lack of high-throughput methods for screening protein regulators of LLPS in target proteins. Here, we developed a CRISPR/Cas9-based screening method to identify protein phase separation regulators by integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly developed method, we screened the RNA-binding proteins that regulate PABPN1 phase separation and identified the tumor suppressor QKI as a promoter of PABPN1 phase separation. Furthermore, QKI exhibits decreased expression levels and diminished nuclear localization in colorectal cancer cells, resulting in reduced PABPN1 phase separation, which, in turn, promotes alternative polyadenylation (APA), cell proliferation, and migration in colorectal cancer.
Humans ; Colorectal Neoplasms/genetics* ; RNA-Binding Proteins/genetics* ; Poly(A)-Binding Protein I/genetics* ; CRISPR-Cas Systems ; Flow Cytometry ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

Objective:To investigate the effects of combination of omalizumab and olofatadine on the improvement of symptoms of refractory chronic spontaneous urticaria (CSU) and its influence on serum immunoglobulin E (IgE) and T cell subsets.Methods:A prospective study was conducted to include 100 patients with refractory CSU who were treated at the Hangzhou Linping District Hospital of Integrated Traditional Chinese and Western Medicine from January 2022 to December 2023. They were divided into the control group and the study group by random number table method. The control group was given olopatadine, 5 mg/ time, 2 times/d, orally; the study group was given the same dose of olofatadine as the control group, plus omazumab, 300 mg/4 weeks, subcutaneous injection. The clinical efficacy of the two groups was compared. The levels of IgE, interleukin-4 (IL-4), interferon -γ (IFN-γ) and T cell subsets before treatment and 12 weeks after treatment were compared between the two groups. The symptom scores and the occurrence of adverse reactions before and after treatment were compared between the two groups.Results:The effective rate in the study group was higher than that in the control group : 96.00% (48/50) vs. 82.00% (41/50), there was statistical difference ( χ2 = 5.01, P<0.05). After treatment, the levels of IgE and IL-4 in the study group were lower than those in the control group : (74.26 ± 12.34) kU/L vs. (93.12 ± 14.62) kU/L, (25.13 ± 3.47) ng/L vs. (29.35 ± 5.24) ng/L, while the level of IFN-γ was higher than that in the control group : (12.72 ± 1.86) ng/L vs. (11.15 ± 1.59) ng/L, there were statistical differences ( P<0.01). After treatment, the level of CD 4+/CD 8+ in the study group was higher than that in the control group 1.50 ± 0.13 vs. 1.01 ± 0.24, there was statistical difference ( P<0.01). After treatment, the scores of refractory CUS symptoms, such as the number of wheals, wheal diameter, pruritus degree and duration in the study group were lower than those in the control group, there were statistical differences ( P<0.05). The total proportion of patients with adverse reactions in the two groups had no statistical difference ( P>0.05). Conclusions:In the treatment of refractory CSU, the combination of omazumab and olatadine can reduce IgE level, improve serum factor indexes, promote the restoration of T cell subpopulation balance, and improve wind mass, pruritosis and other symptoms, with significant clinical efficacy and certain drug safety.

In order to study the pathogenesis of meningitis in lambs caused by Enterococcus faecalis(E.faecalis),and to elucidate the pathogenesis of meningitis from the circRNA level,E.faecalis(named XJ6)was reintroduced into the host to construct a lamb brain tissue injury model,and then the lamb brain tissues were selected for whole genome transcript sequencing at four time in-tervals(24,48,60 and 72 h).Based on the transcriptomics sequencing results,the differential cir-cRNAs were preliminarily screened,and the accuracy of the transcriptomics data was verified by real-time fluorescence quantification.The results showed that the experiments successfully con-structed the brain tissue injury model of E.faecalis infected lambs,and the GO enrichment analy-sis results showed the most significant 10 cellular components,10 biological processes,and 10 mo-lecular functions.KEGG pathway enrichment results revealed that the genes targeted by differenti-al circRNAs were mainly focused on signaling pathways regulating the neuronal cells,the MAPK signaling pathway,the Rap1 signaling pathway,VDEF signaling pathway,Apelin signaling path-way,vascular smooth muscle contraction,and cancer development.The differentially expressed cir-cRNAs validated by real-time quantitative PCR were completely consistent with the transcriptomic results.Among them,novel_circ_0004872 was identified as a candidate circRNA potentially involved in altering the permeability of the blood-brain barrier in lambs,providing a theoretical ref-erence for exploring the regulatory role of circRNAs in blood-brain barrier function.

In order to study the pathogenesis of meningitis in lambs caused by Enterococcus faecalis(E.faecalis),and to elucidate the pathogenesis of meningitis from the circRNA level,E.faecalis(named XJ6)was reintroduced into the host to construct a lamb brain tissue injury model,and then the lamb brain tissues were selected for whole genome transcript sequencing at four time in-tervals(24,48,60 and 72 h).Based on the transcriptomics sequencing results,the differential cir-cRNAs were preliminarily screened,and the accuracy of the transcriptomics data was verified by real-time fluorescence quantification.The results showed that the experiments successfully con-structed the brain tissue injury model of E.faecalis infected lambs,and the GO enrichment analy-sis results showed the most significant 10 cellular components,10 biological processes,and 10 mo-lecular functions.KEGG pathway enrichment results revealed that the genes targeted by differenti-al circRNAs were mainly focused on signaling pathways regulating the neuronal cells,the MAPK signaling pathway,the Rap1 signaling pathway,VDEF signaling pathway,Apelin signaling path-way,vascular smooth muscle contraction,and cancer development.The differentially expressed cir-cRNAs validated by real-time quantitative PCR were completely consistent with the transcriptomic results.Among them,novel_circ_0004872 was identified as a candidate circRNA potentially involved in altering the permeability of the blood-brain barrier in lambs,providing a theoretical ref-erence for exploring the regulatory role of circRNAs in blood-brain barrier function.

Objective:To investigate the effects of combination of omalizumab and olofatadine on the improvement of symptoms of refractory chronic spontaneous urticaria (CSU) and its influence on serum immunoglobulin E (IgE) and T cell subsets.Methods:A prospective study was conducted to include 100 patients with refractory CSU who were treated at the Hangzhou Linping District Hospital of Integrated Traditional Chinese and Western Medicine from January 2022 to December 2023. They were divided into the control group and the study group by random number table method. The control group was given olopatadine, 5 mg/ time, 2 times/d, orally; the study group was given the same dose of olofatadine as the control group, plus omazumab, 300 mg/4 weeks, subcutaneous injection. The clinical efficacy of the two groups was compared. The levels of IgE, interleukin-4 (IL-4), interferon -γ (IFN-γ) and T cell subsets before treatment and 12 weeks after treatment were compared between the two groups. The symptom scores and the occurrence of adverse reactions before and after treatment were compared between the two groups.Results:The effective rate in the study group was higher than that in the control group : 96.00% (48/50) vs. 82.00% (41/50), there was statistical difference ( χ2 = 5.01, P<0.05). After treatment, the levels of IgE and IL-4 in the study group were lower than those in the control group : (74.26 ± 12.34) kU/L vs. (93.12 ± 14.62) kU/L, (25.13 ± 3.47) ng/L vs. (29.35 ± 5.24) ng/L, while the level of IFN-γ was higher than that in the control group : (12.72 ± 1.86) ng/L vs. (11.15 ± 1.59) ng/L, there were statistical differences ( P<0.01). After treatment, the level of CD 4+/CD 8+ in the study group was higher than that in the control group 1.50 ± 0.13 vs. 1.01 ± 0.24, there was statistical difference ( P<0.01). After treatment, the scores of refractory CUS symptoms, such as the number of wheals, wheal diameter, pruritus degree and duration in the study group were lower than those in the control group, there were statistical differences ( P<0.05). The total proportion of patients with adverse reactions in the two groups had no statistical difference ( P>0.05). Conclusions:In the treatment of refractory CSU, the combination of omazumab and olatadine can reduce IgE level, improve serum factor indexes, promote the restoration of T cell subpopulation balance, and improve wind mass, pruritosis and other symptoms, with significant clinical efficacy and certain drug safety.

BACKGROUND:Existing studies have confirmed that exosomes can effectively promote pulp regeneration.However,the biological functions and properties of exosomes from preconditioned sources can be significantly changed,which have different effects on cell proliferation,migration and odontogenic differentiation. OBJECTIVE:To discuss the application status of exosomes and their preconditioning methods in the field of pulp regeneration,and summarize the preconditioning methods that affect the function of exosomes,and explore the effect of exosomes and their preconditioning on pulp regeneration. METHODS:The relevant articles were searched in WanFang,CNKI,PubMed,and Web of Science databases from 2006 to 2022.The Chinese and English search terms were"exosomes,pulp regeneration;preconditioning method".A total of 78 articles were included for analysis. RESULTS AND CONCLUSION:(1)Exosomes have the advantages of good biocompatibility,low immunogenicity and no cytotoxicity,and can induce the regeneration of pulp tissue by promoting stem cell tooth formation,neurogenesis and vascularization.(2)Exosomes derived from preconditioning can enhance the ability of tissue repair and regeneration and have a significant impact on the quality of regenerated dental pulp.(3)Currently,the preconditioning methods used in the field of dental pulp regeneration include inflammatory stimulation,hypoxia induction,conditioned medium and three-dimensional culture,and secreted exosomes can effectively improve the quality of regenerated dental pulp.Nevertheless,the specific effect and mechanism of different preconditioning methods on pulp regeneration need to be explored.

Sepsis-induced liver injury (SILI) is an important cause of septicemia deaths. BaWeiBaiDuSan (BWBDS) was extracted from a formula of Panax ginseng C. A. Meyer, Lilium brownie F. E. Brown ex Miellez var. viridulum Baker, Polygonatum sibiricum Delar. ex Redoute, Lonicera japonica Thunb., Hippophae rhamnoides Linn., Amygdalus Communis Vas, Platycodon grandiflorus (Jacq.) A. DC., and Cortex Phelloderdri. Herein, we investigated whether the BWBDS treatment could reverse SILI by the mechanism of modulating gut microbiota. BWBDS protected mice against SILI, which was associated with promoting macrophage anti-inflammatory activity and enhancing intestinal integrity. BWBDS selectively promoted the growth of Lactobacillus johnsonii (L. johnsonii) in cecal ligation and puncture treated mice. Fecal microbiota transplantation treatment indicated that gut bacteria correlated with sepsis and was required for BWBDS anti-sepsis effects. Notably, L. johnsonii significantly reduced SILI by promoting macrophage anti-inflammatory activity, increasing interleukin-10⁺ M2 macrophage production and enhancing intestinal integrity. Furthermore, heat inactivation L. johnsonii (HI-L. johnsonii) treatment promoted macrophage anti-inflammatory activity and alleviated SILI. Our findings revealed BWBDS and gut microbiota L. johnsonii as novel prebiotic and probiotic that may be used to treat SILI. The potential underlying mechanism was at least in part, via L. johnsonii-dependent immune regulation and interleukin-10⁺ M2 macrophage production.

[This corrects the article DOI: 10.1016/j.apsb.2022.10.016.].

People wearing positive pressure biological protective suits might face some potential safety risks when work -ing in a high-risk biological contamination environment .These risks could be effectively prevented by simulation experi-ments.This paper elaborates on the progress in risk studys on positive pressure biological protective suits in terms of dynam -ic detection methods and risk assessment , and points out the problems to be solved and the direction of future development .

Objective To evaluate the capability of virtual non-contrast (VNC)of the second-generation dual-source CT (DSCT), and to display the adipose tissue of gastric serosa.Methods 48 patients with gastric cancer undergoing preoperative DSCT and sur-gery were enrolled.The true non-contrast (TNC)values and VNC values of perigastric adipose tissue were measured and compared with conventional non-contrast scanning and VNC of arterial phase and venous phase.The VNC values of perigastric adipose tissue were compared between serosal involvement group and non-serosal involvement group.Results There was statistical difference be-tween TNC values and VNC values of perigastric adipose tissue (P <0.05),the absolute differences were under 13 HU.The differ-ence of VNC values between serosal involvement group and non-serosal involvement group was significant (P <0.05).Conclusion VNC of the second-generation DSCT can clearly display the adipose tissue of gastric serosa.It has certain capability in judging sero-sal involvement.VNC has the proficiency to replace conventional non-contrast scanning.