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 molecular mechanism and identify potential drugs for subthreshold depression (SD), and elucidate the detalied mechanism of Danzhi Xiaoyao powder (DZXY) in SD. Methods: Using RNA-sequencing, we identified differentially expressed genes (DEGs) in leukocytes of SD compared to healthy controls, deciphered their functions and pathways, and identified the hub genes of SD. We also assessed changes in leukocyte transcription factor activity in patients with SD using the TELiS platform. The Connectivity Map database was retrieved to screen candidate drugs for SD. Based on network pharmacology, we elucidated the “multi-component, multi-target, and multi-pathway” mechanism of DZXY in the treatment of SD. Results: We identified 1080 DEGs (padj <0.05 and |log2 (fold change)| ≥ 1 & protein coding) in the leukocytes of patients with SD. These DEGs, including hub genes, were primarily involved in immune and inflammatory response-related processes. Transcription factor activity analysis revealed similarities between the leukocyte transcriptome profile in SD and the conserved transcriptional response to adversities in immune cells. Connectivity Map analysis identified 28 potential drugs for SD treatment, particularly SB-202190 and TWS-119. Constructing the “Direct Compounds-Direct Targets-Pathways” network for DZXY and SD revealed the curative mechanisms of DZXY in SD, primarily including inflammatory response, lipid metabolism, immune response, and other processes. Conclusion These results provide new insights into the characteristics and functional changes of leukocytes in SD, partially illustrate the pathogenesis of SD, and suggest potential drugs for SD. The curative mechanisms of DZXY in SD are also partially elucidated.

Objective:To understand the status quo and influencing factors of evidence-based nursing competencies among oncology nurses and provide a reference for enhancing this competency among oncology nurses.Methods:From September 2022 to January 2023, totally 300 practicing nurses from Zhejiang Cancer Hospital were selected by convenience sampling and surveyed with the Chinese version of the Evidence-Based Practice Belief-Knowledge-Practice Scale. Univariate analysis and multiple linear regression were employed to analyze factors affecting the evidence-based nursing competencies of these nurses.Results:The evidence-based nursing competency scores of the 300 oncology nurses were (44.50±10.78) . Among the three dimensions, the belief dimension had the highest average item score, while the practice dimension had the lowest. Multiple linear regression analysis revealed that gender, age, evidence-based training, participation in evidence-based projects, reading professional literature, publication of papers within the last five years and medical statistical level were influencing factors of their evidence-based nursing competencies (all P<0.05) . Conclusions:The evidence-based nursing competencies of oncology nurses are at a relatively low level. Nursing managers and educators should focus on female nurses, older nurses, intensify evidence-based nursing training efforts, and thus enhance the evidence-based nursing competencies and overall research quality of the nursing team.

The stimulator of interferon genes(STING),an integral adaptor protein in the DNA-sensing pathway,plays a pivotal role in the innate immune response against infections.Additionally,it presents a valuable therapeutic target for infectious diseases and cancer.We observed that fangchinoline(Fan),a bis-benzylisoquinoline alkaloid(BBA),effectively impedes the replication of vesicular stomatitis virus(VSV),encephalomyocarditis virus(EMCV),influenza A virus(H1 N1),and herpes simplex virus-1(HSV-1)in vitro.Fan treatment significantly reduced the viral load,attenuated tissue inflammation,and improved survival in a viral sepsis mouse model.Mechanistically,Fan activates the antiviral response in a STING-dependent manner,leading to increased expression of interferon(1FN)and interferon-stimulated genes(ISGs)for potent antiviral effects in vivo and in vitro.Notably,Fan interacts with STING,preventing its degradation and thereby extending the activation of IFN-based antiviral responses.Collectively,our findings highlight the potential of Fan,which elicits antiviral immunity by suppressing STING degra-dation,as a promising candidate for antiviral therapy.

Objective: To clarify the structural and functional characteristics of the gut microbiota in individuals with qi stagnation constitution (QSC) and identify the potential biomarkers related to QSC.Methods: This cross-sectional study involved individuals with QSC and balanced constitution (BC) confirmed by TCM clinicians. The clinical features were recorded, and fecal samples were collected for 16S rDNA sequencing. The structure of the fecal-associated microbiome (FAM) was described by the alpha-diversity indexes, beta-distances, and relative abundances of dominant taxa. The differences in FAM distribution were analyzed by the Wilcoxon rank-sum test, MetagenomeSeq, and LEfSe analysis. The 16S rDNA gene sequences were assigned to the KEGG dataset to predict the functional information of bacterial metabolic pathways by using PICRUSt. Differences in functional pathways between groups were assessed with the Wilcoxon rank-sum test. The ROC curve based on specific operational taxonomic units (OTUs) was constructed, and the AUC was calculated. Results: Twenty-two individuals with BC and 8 with QSC were recruited. Significant differences between the two groups were found in body mass index, health status, and low-density lipoprotein, etc. There was no significant difference in the alpha-diversity index. PCoA showed no evident clustering of bacterial communities according to constitutions. Bacteroidaceae, Lachnospiraceae, Ruminococcaceae, and Pre-votellaceae were the four common bacteria with high abundances. Notably, MetagenomeSeq, LEfSe analysis, and the Wilcoxon rank-sum test identified significantly different distributions of 66, 42, and 36 OTUs, respectively. Predictive function analysis showed that 13 metabolic pathways were significantly differentially distributed, including those related to fatty acid synthesis. Five specific OTUs were selected as potential biomarkers of a QSC, and the AUC was 0.94. Conclusion: Individuals with QSC have unique FAM structure and related functional characteristics. Five specific OTUs were identified to serve as potentially effective biomarkers related to QSC.

OBJECTIVE: To clarify the association between regulatory region of HLA-DPB1 (3'UTR) with Naso pharyngeal carcinoma in Guangdong Province Hans. METHOD: The allelic types of HLA-DPB1-3'UTR were detected by sequence specific primer (SSP) in 104 patients with NPC and 105 control individuals from Guangdong Province Hans. RESULT: The frequencies of allelic types B/B, haplotype B were higher in patients with NPC than those of the control individuals. CONCLUSION Positive association may exist between certain HLA-DPB1 alleles and NPC in Guangdong Province Hans.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Gene Frequency ; HLA-DP Antigens ; genetics ; HLA-DP beta-Chains ; Haplotypes ; Humans ; Molecular Sequence Data ; Nasopharyngeal Neoplasms ; genetics ; Polymorphism, Single Nucleotide

OBJECTIVETo investigate the changes of subgroups of peripheral blood T lymphocytes in patients with severe acute respiratory syndrome (SARS) and its clinical significance.

METHODSSubgroups of blood T lymphocytes in 93 patients with SARS were detected by flow cytometer. The results detected in 64 normal subjects and 50 patients with AIDS served as controls.

RESULTSThe numbers of CD(3)(+), CD(4)(+), and CD(8)(+) lymphocytes all significantly decreased in acute phase of patients with SARS [(722 +/- 533)/microliter, (438 +/- 353)/microliter, (307 +/- 217)/microliter] compared with those in normal controls [(1527 +/- 470)/microliter, (787 +/- 257)/microliter, (633 +/- 280)/microliter, all P <0.01], which was different from what we observed in patients with AIDS who had decreased CD(4)(+) [(296 +/- 298)/microliter] but increased CD(8)(+) [(818 +/- 566)/microliter] counts. The counts of CD(3)(+), CD(4)(+), and CD(8)(+) lymphocytes decreased more apparently in patients with severe SARS. All the five patients who died had CD(4)(+) counts less than 200/microliter. As the patients' condition improved, CD(3)(+), CD(4)(+), and CD(8)(+) counts gradually returned to normal ranges.

CONCLUSIONThe damage of cellular immunity is probably an important mechanism of pathogenesis of SARS.

Acquired Immunodeficiency Syndrome ; immunology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; immunology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo make a comparative study of HLA-DQA1 and HLA-DRB1 allele frequencies in the cases of endometriosis and adeonmyosis.

METHODSThe allelic types of HLA-DQA1 and HLA-DRB1 were detected by polymerase chain reaction-sequence specific primers (PCR-SSP) technique in 51 cases of endometriosis, 45 cases of adenomyosis, and 44 normal individuals as the control.

RESULTSThe frequencies of HLA-DQA1*0401(7.8%, 10.0%) were significantly increased in the endometriosis group and the adenomyosis group (Pc=0.03, Pc=0.01), and the frequencies of HLA-DQA1*0301(8.8%, 5.6%) were significantly decreased in these two groups (Pc=0.00, Pc=0.00).There was no significant difference between the frequencies of HLA-DQA1 and HLA-DRB1 of endometriosis and adenomyosis.

CONCLUSIONThe results indicate that HLA-DQA1*0301 and *0401 alleles are associated with both endometriosis and adenomyosis, and there is perhaps common mechanism involved in both endometriosis and adenomyosis based on HLA-DQA1 and HLA-DRB1 allele frequencies.

Asian Continental Ancestry Group ; genetics ; Endometriosis ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; HLA-DQ Antigens ; genetics ; HLA-DQ alpha-Chains ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Polymerase Chain Reaction ; methods

By making investigation on the animals with venom protein and peptied.In China sea we find that Coelenterata of them mainly includes 14 kinds of Jellyfist,15 kinds of Sea anemone,one kind of Corals and two kinds of Hydrozoa,altogether 32 kinds;Mollusea of them mainly includes 34 kinds of Conus,two kinds of Aplysia and 3 kinds of Cephalopoda,altogether 39 kinds;Nemartina of them has one kind;Echinodermata of them mainly includes two kinds of starfish and 8 kinds of Sea urchins,altogether 10 kinds;fishes of them mainly include one kind of Venomous fishes,two kinds of Ichthyocrinotoxic fishes and 74 kinds of Venomous fishes,altogether 77 kinds;Sea snake is 15 kinds.These animals with venom protein and peptide mainly are distributed in South sea of China.

Objective Vessel injury provokes the release of angiotensin II that influences the proliferation and migration of vascular smooth muscle cells (VSMCs). T lymphocytes produced, interleukin 10 is of immunoregulatory function. The purpose of this study is to determine the effects of recombinant human interleukin 10 (rhIL 10) on the proliferation of isolated rat vascular smooth muscle cell and the activity of p44/p42 mitogen activated protein kinase (MAPK) promoted by angiotensin II. Methods Rat aortic VSMCs were cultured and treated with rhIL 10 or angiotensin II, and then co treated with rhIL 10 and angiotensin II. The proliferation was quantified by colormetric assay. The p44/p42 MAPK activity was evaluated by the immunobloting technique using anti p44/p42 phospho MAPK antibody. Results Compared with control group, angiotensin II stimulated significantly VSMCs proliferation(1 311?0 201 vs 0 781?0 236, P 0 05), but significantly inhibited VSMCs proliferation induced by angiotensin II at a dose of 1,10,100 ng/ml (0 984?0 172 , 0 932?0 134 , 0 784?0 097 vs 1 311?0 201, P