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

The fungal bioluminescence pathway (FBP) catalyzes the oxidation of endogenous caffeic acid to produce green bioluminescence through an enzymatic cascade. Genetic engineering of FBP into plants creates autoluminescent specimens that circumvent the substrate limitations of conventional reporter systems. These transgenic plants serve dual functions as aesthetic displays and versatile biosensing platforms, enabling applications in real-time gene expression monitoring, continuous environmental surveillance, and non-invasive bioimaging, offering novel opportunities for horticultural production, environmental conservation, and bioengineering applications. This review synthesizes current advances in plant FBP engineering and explores how machine learning approaches can optimize autoluminescent phenotypes, thereby accelerating innovation in agricultural biotechnology, environmental sensing, and synthetic biology applications.
Fungi/genetics* ; Plants, Genetically Modified/metabolism* ; Genetic Engineering ; Biosensing Techniques ; Luminescent Measurements ; Caffeic Acids/metabolism* ; Luminescence

Objective To develop a diagnostic model combining the CT angiography(CCTA)-derived myocardial radiomics signatures with the CT-derived fractional flow reserve(CT-FFR)based on coronary CCTA and investigate the diagnostic accuracy of the hybrid model for hemodynamically significant coronary artery disease(CAD).Methods The patients presenting stable angina pectoris,diagnosed with CAD,and clinically referred for CCTA examination and invasive coronary angiography were prospectively recruited.Radiomics features of the left ventricular myocardium were extracted from the three main perfusion territories demarcated according to the coronary blood supply.The extracted features were first selected by the minimum redundancy maximum relevance feature ranking method.A least absolute shrinkage and selection operator Logistic regression algorithm with leave-one-out cross-validation was then employed to construct a radiomics model.The CT-FFR value was generated for each blood vessel.The area under the receiver operating characteristics curve(AUC_ROC),sensitivity,and specificity were adopted to evaluate the performance of each model against the reference standard invasive coronary angiography/FFR.Results A total of 70 patients[42 men and 28 women;(61±10) years old] were included in this study and complemented CCTA examination,with 175 vessels and the corresponding myocardial territories undergoing invasive coronary angiography/FFR.A total of 1 656 specific radiomics parameters were extracted,from which 14 features were selected to establish the radiomics model.The AUC_ROC,sensitivity,and specificity were 0.797(95%CI=0.732-0.861),77.1%,and 73.7%for the radiomics model,0.892(95%CI=0.841-0.943),81.4%,and 88.8%for the CT-FFR model,and 0.928(95%CI=0.890-0.965),83.3%,and 88.4%for the hybrid model,respectively.The hybrid model outperformed the radiomics model and CT-FFR alone(P=0.040).Conclusions The radiomics signatures of the vessel-related myocardium from CCTA could provide incremental value to the diagnostic performance of CT-FFR and improve vessel-specific ischemia detection.The hybrid model combining CT-FFR with radiomics signatures is potentially feasible for improving the diagnostic accuracy for hemodynamically significant CAD.
Coronary Angiography/methods* ; Tomography, X-Ray Computed ; Humans ; Hemodynamics ; Coronary Artery Disease/diagnostic imaging* ; Male ; Female ; Middle Aged ; Aged ; Radiomics ; Angina Pectoris/diagnostic imaging* ; China ; Image Processing, Computer-Assisted ; Coronary Vessels/diagnostic imaging*

Objective Bevacizumab has been clinically used in colorectal cancer,ovarian cancer,cervical cancer,non-small cell lung cancer and other tumor diseases.Common adverse reactions during bevacizumab treatment include albuminuria,thrombosis,bleeding,gastrointestinal perforation and hypertension,among which the incidence of hypertension is as high as 19％-47％.The occurrence of hypertension affects the quality of life of patients,hinds the normal development of tumor treatment,and even induces serious cardiovascular diseases and increases the risk of death,which requires clinical attention.In this paper,the mechanism,influencing factors,prognosis and related treatment of bevacizumab associated hypertension were reviewed,so as to provide reference for clinical rational drug use.

Objective: To investigate the immune reconstitution of HIV/AIDS patients and its influencing factors after receiving antiviral therapy (ART) in Hangzhou City, so as to provide insights into improving the treatment effects and quality of life in HIV/AIDS patients. Methods: A retrospective cohort of HIV/AIDS patients who began antiviral treatment between January 1, 2016 and August 31, 2021 and had a baseline CD4+T lymphocyte (CD4) counts of less than 500 cells/μL or a baseline CD4/CD8+T lymphocyte (CD8) ratio of less than 0.8 in Hangzhou City was followed up until August 31, 2023. Demographic information, antiviral therapy in formation, CD4 counts, and CD4/CD8 were collected from the Chinese Disease Prevention and Control Information System. A good immune reconstitution was defined as having CD4≥500 cells/μL and CD4/CD8≥0.8. The immune reconstitution status of HIV/AIDS patients were analyzed, and factors affecting immune reconstitution were identified using a multivariable Cox proportional risk regression model. Results: A total of 3 349 HIV/AIDS patients were enrolled, with a median age at ART of 31 (interquartile range, 20) years. There were 3 075 males (91.82%), 1 600 cases with college education and above (47.78%) and 2 455 cases at WHO clinical stage Ⅰ-Ⅱ(73.31%). There were 1 368 cases with good immune reconstitution, accounting for 40.85%, and the proportion of HIV/AIDS patients with good immune reconstitution that began ART in 2016 was the highest, reaching 51.90%. Multivariable Cox proportional risk regression model identified WHO clinical stage (Ⅰ-Ⅱ, HR=2.529, 95%CI: 2.023-3.162), timely ART (HR=1.196, 95%CI: 1.027-1.394), initial treatment regimen (TDF+3TC+NVP/EFV, HR=2.185, 95%CI: 1.891-2.524; integrase inhibitors, HR=8.509, 95%CI: 6.706-10.795), baseline CD4/CD8 (≥0.1, HR: 1.600-4.515, 95%CI: 1.061-6.661), baseline hemoglobin (<90 mg/dL, HR=0.327, 95%CI: 0.121-0.880), hepatitis B infection (HR=0.619, 95%CI: 0.457-0.840) and hepatitis C infection (HR=0.308, 95%CI: 0.099-0.956) as factors affecting immune reconstitution in HIV/AIDS patients. Conclusion The immune reconstitution in HIV/AIDS patients after ART is associated with WHO clinical stage, timely ART, initial treatment regimen, baseline CD4/CD8, baseline hemoglobin and hepatitis B or C infection.

italic>Anoectochilus roxburghii liquid (spray, a hospital preparation of Wu Mengchao Hepatobiliary Hospital of Fujian Medical University) has shown a good clinical treatment effect during the COVID-19 pandemic, but its material basis and mechanism of action are still unclear. In this study, network pharmacology and molecular docking methods were used to predict the molecular mechanism of A. roxburghii liquid against COVID-19, and pharmacodynamic experiments in vitro were conducted to study the interaction between the current targets with clear preventive and therapeutic effects and the key components of A. roxburghii liquid. UPLC-MS and database were used to compare and analyze the active ingredients in the liquid, and 17 potential active ingredients with good drug-like properties were screened by in vivo pharmacokinetics process in SwissADME database. SwissTargetPrediction and GeneCards were searched to find 93 common targets. Cytoscape 3.8.2 software was used to construct the "component-target" network map, and the Metascape platform was used for gene function annotation and pathway enrichment analysis. It was found that the extract could regulate the positive response to external stimuli, inflammatory response, cytokine production and other biological processes by binding the active ingredients such as isorhamnetin, kaempferol, luteolin, quercetin and apigenin to the common targets (NOS3, MPO, MMP3, etc.), and play an anti-COVID-19 role. In the angiotensin-converting enzyme 2 (ACE2) activity inhibition assay, it was found that the stock solution of A. roxburghii liquid (for spray), and the supernatant after removing polysaccharides (mainly containing flavonoids) could to some extent inhibit the activity of ACE2. Crucially, in the experiment of 2019-nCOV-S pseudovirus infecting HEK-293T-ACE2 cells, we found that A. roxburghii liquid may exert anti-COVID-19 effects by blocking the binding of SARS-CoV-S protein to ACE2.

Objective To investigate the toxicokinetic differences of 3,4-methylenedioxy-N-methylamphetamine(MDMA)and its metabolite 4,5-methylene dioxy amphetamine(MDA)in rats af-ter single and continuous administration of MDMA,providing reference data for the forensic identifica-tion of MDMA.Methods A total of 24 rats in the single administration group were randomly divided into 5,10 and 20 mg/kg experimental groups and the control group,with 6 rats in each group.The ex-perimental group was given intraperitoneal injection of MDMA,and the control group was given intraperi-toneal injection of the same volume of normal saline as the experimental group.The amount of 0.5 mL blood was collected from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.In the continuous administration group,24 rats were randomly divided into the experi-mental group(18 rats)and the control group(6 rats).The experimental group was given MDMA 7 d by continuous intraperitoneal injection in increments of 5,7,9,11,13,15,17 mg/kg per day,respectively,while the control group was given the same volume of normal saline as the experimental group by in-traperitoneal injection.On the eighth day,the experimental rats were randomly divided into 5,10 and 20 mg/kg dose groups,with 6 rats in each group.MDMA was injected intraperitoneally,and the con-trol group was injected intraperitoneally with the same volume of normal saline as the experimental group.On the eighth day,0.5 mL of blood was taken from the medial canthus 5 min,30 min,1 h,1.5 h,2 h,4 h,6 h,8 h,10 h,12 h after administration.Liquid chromatography-triple quadrupole tandem mass spectrometry was used to detect MDMA and MDA levels,and statistical software was employed for data analysis.Results In the single-administration group,peak concentrations of MDMA and MDA were reached at 5 min and 1 h after administration,respectively,with the largest detection time limit of 12 h.In the continuous administration group,peak concentrations were reached at 30 min and 1.5 h af-ter administration,respectively,with the largest detection time limit of 10 h.Nonlinear fitting equations for the concentration ratio of MDMA and MDA in plasma and administration time in the single-administration group and continuous administration group were as follows:T=10.362C-1.183,R2=0.974 6;T=7.397 3C-0.694,R2=0.961 5(T:injection time;C:concentration ratio of MDMA to MDA in plasma).Conclusions The toxicokinetic data of MDMA and its metabolite MDA in rats,obtained through single and continuous administration,including peak concentration,peak time,detection time limit,and the relationship between concentration ratio and administration time,provide a theoretical and data foundation for relevant forensic identification.

Objective To study the expression of amplified in breast cancer 1(AIB1)and ankyrin repeat domain containing11(ANKRD11/ANCO1)in human papillomavirus(HPV)positive cervical cancer and their relationship with clinicopathological characteristics and their predictive value for postoperative recurrence.Methods Cancer tissues and paracancer tissues of 94 HPV positive cervical cancer patients who visited the Third People's Hospital of Mianyang from January 2018 to January 2020 were selected,while 50 HPV negative cervical cancer tissues during this period were taken as controls.Real-time qPCR(RT-qPCR)was conducted to detect AIB1 mRNA and ANCO1 mRNA expression in cancer tissue and paracancer tissues of HPV positive cervical cancer and cancer tissue of HPV negative cervical.Pearson analysis of the correlation between AIB1 mRNA and ANCO1 mRNA expression in HPV positive cervical cancer tissues.Logistic regression analysis was used to identify the risk factors for postoperative recurrence of HPV-positive cervical cancer patients.The predictive value of AIB1 mRNA,ANCO1 mRNA,and their combination in predicting postoperative recurrence in HPV positive cervical cancer patients was analyzed by receiver operating characteristic curve.Results Compared to the adjacent tissues of HPV positive cervical cancer and cancer tissues of HPV negative cervical cancer,AIB1 mRNA(3.04±0.37 vs 0.87±0.21,1.02±0.33)in HPV positive cervical cancer tissues was higher,while ANCO1 mRNA(1.13±0.26 vs 1.91±0.35,1.82±0.36)was lower,with significant differences(t=68.499,53.137;23.649,17.434,all P<0.05).The expression of AIB1 mRNA and ANCO1 mRNA in HPV positive cervical cancer tissues showed a negative correlation(r=-0.714,P<0.001).Compared to patients with FIGO stage ⅡA～ⅡB and without lymph node metastasis,AIB1 mRNA(3.88±0.32 vs 2.04±0.41,4.46±0.33 vs 2.16±0.46)in HPV positive cervical cancer tissues with FIGO stage ⅡA and lymph node metastasis was higher,while ANCO1 mRNA(0.67±0.29 vs 1.68±0.20,0.49±0.24 vs 1.53±0.32)was lower,with significant differences(t=24.425,26.097;19.288,16.777,all P<0.001).FIGO stage ⅡA,lymph node metastasis,and high AIB1 mRNA[OR(95％CI)=1.644(1.223～2.210),1.779(1.295～2.444),1.247(1.050～1.728)]were risk factors for postoperative recurrence in HPV positive cervical cancer patients,while high ANCO1 mRNA[(OR(95％CI):0.634(0.451～0.891)]was a protective factor.The AUC(95％CI)of AIB1 mRNA and ANCO11 mRNA combined for predicting postoperative recurrence in HPV positive cervical cancer patients was 0.914(0.863～0.952),which was higher than the single indicator detection of 0.821(0.782～0.869)and 0.794(0.763～0.847),and the differences were significant(Z=4.123,4.432,all P<0.001).Conclusion The expression of AIB1 mRNA is increased and the expression of ANCO1 mRNA is reduced in HPV positive cervical cancer tissue,which are related to the occurrence and progression of cancer.The combination of the two has a high predictive value for evaluating postoperative recurrence.

Objective:To understand the serotype distribution, drug resistance and molecular characterization of invasive non-typhoid Salmonella (iNTS) in Guangdong Province from 2018 to 2022 and provide scientific evidence for the prevention and treatment of blood flow infection caused by Salmonella. Methods:Serological identification, antimicrobial susceptibility testing, multilocus sequence typing (MLST), and whole genome sequencing were performed on Salmonella isolated from blood and stool samples in Guangdong from 2018 to 2022. Simultaneously, annotated the sequencing results for drug resistance genes and virulence factors by a microbial gene annotation system. Results:The 136 iNTS strains were divided into 25 serotypes, and Salmonella enteritidis accounted for 38.24% (52/136). The OR of other iNTS serotypes were calculated with Salmonella typhimurium as the control. The OR values of Oreninburg, Rysson, and Pomona serotypes were the highest, which were 423.50, 352.92, and 211.75, respectively. The drug resistance rate of iNTS was 0.74%-66.91%, which was lower than that of non-iNTS (3.90%-77.21%). The main iNTS of drug resistance were ampicillin and tetracycline, with resistance rates of 66.91% (91/136) and 50.00% (68/136), respectively, while the resistance rates to ciprofloxacin (5.88%,8/136), ceftazidime (5.88%,8/136), gentamicin (5.13%,7/136) and cefoxitin (0.74%, 1/136) were relatively low. iNTS carried a variety of drug-resistance genes and virulence factors, but no standard virulence factor distribution has been found. MLST cluster analysis showed that iNTS was divided into 26 sequence types, and ST11 accounted for 38.24% (52/136). Conclusions:The iNTS strains in Guangdong were dominated by Salmonella enteritidis, of which three serotypes, Oreninburg, Rison, and Pomona, may be associated with a higher risk of invasive infection during 2018 to 2022 . iNTS was sensitive to clinical first-line therapeutic drugs (cephalosporins and fluoroquinolones), with highly diverse sequences and clear phylogenetic branches. ST11 was the local dominant clone group.