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 observe the effect of combining heat-sensitive moxibustion with action observation therapy in the treatment of spastic wrist and hand dysfunction post-stroke.Methods:Ninety stroke survivors with spasmodic wrist and hand dysfunction were randomly assigned to a heat-sensitive moxibustion group, an action observation group, or a combination group, each of 30. In addition to routine rehabilitation treatment, the groups received heat-sensitive moxibustion, action observation therapy or moxibustion followed by action observation therapy in 6 sessions a week over 6 weeks. Surface electromyography signals (iEMG and RMS) were recorded from the affected wrist and hand muscles before and after the treatment. All of the participants were also assessed using the Fugl-Meyer motor assessment for the upper extremities (FMA-UE).Results:After treatment, a significant improvement was observed in the average FMA-UE values, with that of the combination group (17.2±2.1) significantly higher than those of the other two groups. The average iEMG of the wrist flexors and extensors, the extensors of the fingers, and abductor pollicis brevis had increased significantly, with the combination group′s averages significantly higher than those of the other two groups. The average RMS values of those muscles had decreased significantly after the treatment, with those of the combination group significantly lower than the others.Conclusions:Combining heat-sensitive moxibustion and action observation therapy can significantly reduce muscle tension, and promote strength and better functioning of spastic wrist and hand muscles after a stroke. The combination is worthy of clinical application and promotion.

Objective To investigate the relationship between tooth loss and the occurrence of esophageal cancer in a natural village in Wenfeng District, Anyang City, Henan Province. Methods A prospective cohort study was conducted to observe the occurrence of tooth loss and esophageal cancer among the asymptomatic residents of the natural village for 16 years from January 2008 to July 2024. Data were analyzed by chi-square test, binary logistic regression, and restricted cubic spline. Results Among the total population of 711 cases, 136 cases were lost to follow-up and 575 cases were included in the final statistics, including 45 cases with esophageal cancer. Significant statistical difference was found between esophageal cancer patients with and without tooth loss (P<0.05). Logistic regression analysis showed that tooth loss was associated with the occurrence of esophageal cancer (OR=3.977, 95%CI: 1.543-10.255). After the adjustment for confounders, tooth loss remained significantly associated with the occurrence of esophageal cancer (OR=3.038, 95%CI: 1.035-8.914). A nonlinear dose-response relationship was observed between the number of teeth lost and the incidence of esophageal cancer. When the number of teeth lost was less than 12, the risk of esophageal cancer increased with the number of teeth lost. When more than 12 teeth were lost, the risk of esophageal cancer decreased as the number of teeth lost increased. Conclusion Tooth loss is a risk factor for the occurrence of esophageal cancer in this natural village. When the number of teeth lost is less than 12, the risk of esophageal cancer increases with the number of teeth lost.

Objective To investigatethe relationship between gastroesophageal reflux disease (GERD) symptoms and clinicopathological characteristics, p53 expression, and survival of Chinese patients with esophageal adenocarcinoma. Methods A total of 3882 patients with esophageal adenocarcinoma, 970 matched patients with esophageal squamous cell carcinoma, and 970 matched patients with gastric cardia adenocarcinoma were included to compare the incidence of GERD symptoms. Patients with esophageal adenocarcinoma were divided into two groups according to the presence and absence of GERD symptoms. The differences in clinicopathological characteristics and p53 expression between the two groups were compared by chi-square test, and the differences in survival between the two groups were compared by landmark analysis. Results The incidence of GERD symptoms increased successively in esophageal squamous cell carcinoma, esophageal adenocarcinoma, and gastric cardia adenocarcinoma. In patients with esophageal adenocarcinoma, the proportions of male, less than 65 years old, lower thoracic tumors, tumors without squamous cell carcinoma, poorly differentiation, early tumors (stage 0-I), and p53-positive tumors in the group with GERD symptoms were higher than those in the group without GERD symptoms; moreover, the long-term survival (≥15 years) was better. Conclusion GERD symptom is an important clinical sign of esophageal adenocarcinoma. It is closely related to specific clinicopathological characteristics, p53 overexpression, and good long-term prognosis.

Objective:To investigate the association of the interaction and combined effect of renal function and body mass index (BMI) with the risk for all-cause death in patients with type 2 diabetes mellitus (T2DM) in communities of Jiangsu Province.Methods:The study subjects were from the Comprehensive Research Project of Diabetes Prevention and Control conducted in Jiangsu from December 2013 to January 2014, and follow up was conducted for them until September 30, 2023. A total of 20 025 subjects were included in the study. Cox proportional hazards regression model was used to analyze the association of renal function with risk for death in T2DM patients, and the association of interaction between renal function and BMI and their combined effect with all-cause death risk in T2DM patients.Results:In the follow up for 198 370 person-years, a total of 4 459 deaths were recorded. Cox proportional hazards regression model analysis showed that renal dysfunction was associated with 71% risk of all-cause mortality in all T2DM patients [hazard ratio ( HR) =1.71, 95% CI: 1.59-1.84], as well as in all BMI subgroups. Likelihood ratio test indicated an interaction between renal function and BMI (interaction for P=0.030). Compared with patients with normal renal function and normal BMI, those with normal renal function and over weight or obesity had a lower risk of all-cause mortality, and those with renal dysfunction and low weight had the highest risk for death ( HR=2.78, 95% CI: 1.87-4.14). Conclusions:There is association of interaction between renal function and BMI with all-cause mortality in T2DM patients. T2DM patients with renal dysfunction and low body weight had significant higher risk for death.

AIM:This study aimed to utilize network pharmacology and an anxiety rat model to investigate the intervention targets and underlying mechanisms of Baixiangdan capsule(BXD)in the treatment of anxiety-like behavior.METHODS:We screened the action targets of BXD on the anxiety model using network pharmacology,followed by Gene Ontology(GO)enrichment and the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis of the identified targets.Additionally,we evaluated the effects of BXD on rat anxiety behaviors induced by foot shock,with measurement of serum level of c-Jun N-terminal kinase inhibitor(JIK),and the protein levels of c-Jun N-terminal kinase(JNK),phos-phorylated JNK(p-JNK)and JIK in the left hippocampus and hypothalamus.RESULTS:A total of 97 targets,including the dopamine D1/D3 receptors,were identified as intersection targets.GO enrichment analysis indicated that target pro-teins were significantly involved in molecular functions such as catecholamine binding,receptor activation,and signal transduction.KEGG pathway analysis revealed that neural receptor-ligand pathways,including dopaminergic synapses(15%)and neuroactive ligand-receptor interaction(18%),were notably represented.In behavioral experiments,BXD and diazepam treatments significantly reduced total path in the open-field test(P＜0.01),while the number of entries into the central region increased(P＜0.05).Additionally,the percentage of entry times of the open arm and the percentage of time spent in the open arm were also increased(P＜0.01).Furthermore,BXD treatment led to decreased ratio of p-JNK/JNK and increased level of JIK(P＜0.05).CONCLUSION:The traditional Chinese medicine BXD demonstrates a promising efficacy in alleviating anxiety-like behaviors in model rats.It operates through a multi-component,multi-target,and multi-pathway approach,primarily by modulating the JIK/JNK signaling pathway.

Purpose To investigate the correlation between the methylation level at CpG sites of CXCL17 and the clinicopathological parameters of papillary thyroid carcinoma(PTC).Methods samples from 186 cases of PTC and 191 cases of benign thyroid nodule(BTN)were collected.Methylation levels of CXCL17 were semi-quantitatively as-sessed using mass spectrometry.Logistic regression analysis,which adjusted for age,gender and related hormones,was conducted to evaluate the correlation between CXCL17 methylation and PTC,and calculate the odds ratios(ORs)and 95％confidence intervals(CIs).Results Hypomethylation level of CXCL17_CpG_1.2 was significantly associat-ed with PTC(OR=1.36,95％CI:1.16-1.60,P＜0.001)and early stage of PTC patients(Stage Ⅰ,OR=1.41,95％CI:1.19-1.67,P＜0.001).Gender-based hierarchical management analysis showed that decreased methyla-tion level of CXCL17_CpG_1.2 was significantly associated with female PTC patients(OR=1.39,95％CI:1.15-1.67,P＜0.001).In subgroups stratified by age(＜50 and≥50 years old),hypomethylation at CXCL17_CpG_1.2 was significantly associated with PTC,with a stronger association in the younger subgroup(＜50 years old:OR=1.42,95％CI:1.14-1.77,P＜0.01;≥ 50 years old:OR=1.30,95％CI:1.03-1.64,P＜0.05).Conclusion There was a significant difference in CXCL17 methylation levels between benign and malignant thyroid tumors.It was showed that hypomethylation of CXCL17 is closely associated with PTC,particularly in young women patient.Thus,CXCL17 methylation may serve as a biomarker for accurate differential diagnosis of thyroid nodule.

A case(male,17 years old)of anterior teeth with grade Ⅲ open bite,Class Ⅱ malocclusion and high facial angle was trea-ted with palatal mini-screw implant anchorage and high-pull headgear after extraction of 4 second premolars,bilateral maxillary second molars and bilateral mandibular third molars.The vertical height was controlled,the mandible was rotated counterclockwise.The sagittal and vertical facial growth pattern was improved and an optimal occlusal relationship was achieved.

Objective:To study the allocation efficiency of private ophthalmology health resources in Shanxi,and to provide references for improving the allocation efficiency of health resources in Chinese private ophthalmology medical institutions.Methods:The resource allocation and services of 70 private ophthalmic medical institutions in Shanxi were collected through a questionnaire survey,and Data Envelopment Analysis(DEA)was used to evaluate the efficiency of health resource allocation in medical institutions of Shanxi.Results:The average values of technical efficiency,pure technical efficiency,and scale efficiency of health resource allocation in private ophthalmic medical institutions in Shanxi were 0.963,0.980,and 0.982,respectively.Among the 70 private ophthalmology institutions,7 institutions were DEA-strongly efficient in health resource allocation,26 institutions were DEA-weakly efficient,37 institutions were non-DEA efficient,15 institutions had constant return to scale,40 institutions had increasing return to scale,and 15 institutions had decreasing return to scale.The allocation of health resources in 7 cities,including Taiyuan,Datong,and Shuozhou,etc.were DEA-strongly efficient;Changzhi and Jincheng were DEA-weakly efficient,both with increasing return to scale;and Linfen was non-DEA efficient with increasing return to scale.Conclusion:The efficiency of health resource allocation in some municipalities of Shanxi needs to be improved;the level of inter-organization varied,and the problems of insufficient resources and wasted inputs coexisted.In the future,ophthalmic resources should be rationally allocated,and input and output indicators should be adjusted according to the actual situation.

Objective:To investigate the expression and methylation status of the SALL4 gene in placental tissues of fetal growth restriction (FGR) and its effects on trophoblast cell proliferation, migration, and invasion. Methods:Placental tissues were collected from 20 full-term FGR patients and 20 healthy term controls who underwent regular prenatal examination and cesarean section at the Third Affiliated Hospital, Zhengzhou University between July 2023 and February 2024. SALL4 mRNA and protein expression were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Methylation specific polymerase china reaction (MSP) assessed promoter methylation levels. HTR8/SVneo cells were transfected with SALL4-targeting small interfering RNA (si-SALL4) or negative control small interfering RNA (si-NC). HTR8/SVneo cells were treated with the demethylating agent 5-aza-2′-deoxycytidine (5-Aza-dC) to inhibit gene methylation (5-Aza-dC group) or with 10% RPMI-1640 medium as a vehicle control. Transfection efficiency (for siRNA) and the efficacy of 5-Aza-dC-induced demethylation were assessed by qRT-PCR and Western blot. The functional effects of SALL4 knockdown and methylation inhibition on trophoblast cells were evaluated using proliferation assays, scratch wound healing assays, and Transwell invasion assays. Statistical analyses included independent t-tests and Chi-square test. Results:(1) Human tissues: FGR placentas showed lower SALL4 mRNA (0.802±0.194 vs. 1.015±0.186, t=3.55) and protein expression (0.445±0.114 vs. 0.701±0.113, t=3.19), alongside higher methylation rates of SALL4 [80% (16/20) vs. 15% (3/20), χ2=14.44] compared to controls (all P<0.05). (2) In vitro: si-SALL4 transfection reduced HTR8/SVneo proliferation (OD450 at 48 h: 0.653±0.021 vs. 0.827±0.040, t=6.60), migration [healing rate at 48 h: (24.317±2.637)% vs. (49.327±1.961)%, t=13.18], and invasion [counted invaded cells: (133.000±6.557) vs. (272.667±18.009) cells, t=12.62] versus si-NC (all P<0.05). Conversely, 5-Aza-dC treatment increased HTR8/SVneo proliferation (0.917±0.042 vs. 0.783±0.031, t=－4.47), migration [(71.097±3.354)% vs. (51.632±2.877)%, t=－7.63], and invasion [(384.000±12.166) vs. (202.833±7.095) cells, t=－13.69] versus vehicle control (all P<0.05). Conclusions:Hypermethylation of the SALL4 promoter in FGR placentas suppresses its expression, impairing trophoblast cell function. Demethylation restores SALL4 expression and enhances cellular proliferation, migration, and invasion, involving in the occurrence and development of FGR disease.