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 hypertension (HTN) trends, key risk factors, and gender disparities in rural China, and to propose targeted strategies for improving HTN control in resource-limited settings. METHODS: This longitudinal study used data from the Henan Rural Cohort Study, including baseline (2015-2017; n = 39,224) and follow-up (2018-2022; n = 28,621) participants. HTN was defined as systolic/diastolic blood pressure ≥ 140/90 mmHg, self-reported diagnosis, or use of antihypertensive medication. Severity was classified using a 7-tier blood pressure (BP) staging system (optimal, normal, high normal, and HTN stages 1-4). A generalized linear mixed-effects model (GLMM) identified associated risk factors. RESULTS: HTN prevalence increased modestly from 32.7% (95% CI: 32.2-33.2) to 33.9% (95% CI: 33.3%-34.4%). Awareness and treatment improved from 20.1% to 25.3%, and from 18.8% to 24.4%, respectively, but control rates remained low (6.2% to 12.3%). After adjustment, women had a 1.53-fold higher HTN risk than men ( OR = 1.53, 95% CI: 1.43-1.63), revealing gender-specific trends. Key risk factors included alcohol use ( OR = 1.37, 95% CI: 1.27-1.47) and overweight status ( OR = 1.76, 95% CI: 1.66-1.86). BP staging showed an increase in optimal BP (42.3% to 45.8%), but stagnant management of advanced HTN stages. CONCLUSION Hypertension in rural China is shaped by behavioral risk factors and healthcare access gaps. Gender-sensitive, community-based interventions, including task-shifting models, are necessary to mitigate the growing burden of hypertension.
Humans ; Hypertension/etiology* ; China/epidemiology* ; Female ; Male ; Rural Population/statistics & numerical data* ; Prevalence ; Risk Factors ; Middle Aged ; Adult ; Aged ; Longitudinal Studies ; Sex Factors ; Cohort Studies ; East Asian People

Objective To investigate the safety and efficacy of totally no tube three-port thoracoscopic surgery (TNTT) for thymic tumor via lateral thoracic approach. Methods The clinical data of patients with thymoma admitted to the Department of Thoracic Surgery of the General Hospital of Northern Theater Command from November 2021 to May 2022 were retrospectively analyzed. The patients were divided into a TNTT group and a single utility port video-assisted thoracic surgery (SVATS) group according to different surgical methods. The clinical data were compared between the two groups. Results A total of 111 patients were collected. There were 44 patients in the TNTT group, including 20 males and 24 females, with an average age of 60.11±8.64 years, and 67 patients in the SVATS group, including 30 males and 37 females, with an average age of 62.40±7.92 years. There was no significant difference between the two groups in the baseline data (P>0.05). The postoperative hospital stay and intraoperative blood loss were shorter or less in the TNTT group (P<0.05), and the visual analogue scale score 48 hours after the operation was smaller in the SVATS group (P<0.05). Conclusion TNTT has a good surgical safety, and can shorten postoperative hospital stay, reduce intraoperative blood loss, and has significant advantages in enhanced recovery after surgery, but SVATS can reduce postoperative pain in patients.

Objective To analyze the learning curve of Da Vinci robotic segmentectomy. Methods Cumulative sum analysis (CUSUM) was used to analyze the learning curve of Da Vinci robotic segmentectomy performed by the General Hospital of Northern Theater Command from February 2018 to December 2020. The learning curve was obtained by fitting, and R2 was used to judge the goodness of fitting. The clinical data of patients in different stages of learning curve were compared and analyzed. Results The first 50 patients who received Da Vinci robotic segmentectomy were included, including 24 males and 26 females, with an average age of 61.9±10.6 years. The operation time decreased gradually with the accumulation of operation patients. The goodness of fitting coefficient reached the maximum value when R2=0.907 (P<0.001), CUSUM (n) =0.009×n3−0.953×n2+24.968×n−7.033 (n was the number of patients). The fitting curve achieved vertex crossing when the number of patients reached 17. Based on this, 50 patients were divided into two stages: a learning and improving stage and a mastering stage. There were statistical differences in the operation time, intraoperative blood loss, postoperative drainage volume, number of lymph node dissection, postoperative catheter time, postoperative hospital stay, and postoperative complications between the two stages (P<0.05). Conclusion It shows that the technical competency for assuring feasible perioperative outcomes can be achieved when the cumulative number of surgical patients reaches 17.

Objective:To investigate effects of polydimethylsiloxane coating on the quality characteristics and qual-ity control methods of medium borosilicate glass tube injection bottles,including appearance,identification,physi-cal properties,water resistance of inner surface and element leaching amount.Methods:Nine batches of coated medium borosilicate glass tube injection bottles and corresponding medium borosilicate glass tube injection bottles from three companies were used to evaluate whether the quality control method of medium borosilicate glass tube in-jection bottles after polydimethylsiloxane film was modified,and to evaluate the effect of coating on the quality char-acteristics of bottles.Results:The coating did not have effect on appearance,identification and physical properties of medium borosilicate glass tube injection bottle.The coating process did not damage the original performance of the medium borosilicate glass tube injection bottle,and lead to the matters with special attention on the inspection method.The water resistance of the inner surface of the coated medium borosilicate glass tube injection bottle was improved.The leaching amounts of harmful elements such as arsenic,antimony,lead and cadmium before and after coating were low,indicating that the coating process did not increase the leaching risk of these harmful elements.The leaching amounts of elements were determined by ICP-OES,and no significant changes were found in most metal elements.Conclusion:The appearance,identification,physical properties and element leaching quality con-trol method of medium borosilicate glass tube injection bottle after poly dimethylsiloxane film coating do not need to be changed.The water resistance of the inner surface after coating is improved,and there is no significant adverse effect on other quality characteristics.

Objective:To investigate the correlation between RYR1 gene and the development of gastric cancer,as well as the mechanism of RYR1 in promoting the progression of gastric cancer.Methods:We analyzed gastric cancer data from TCGA and conducted high-throughput targeted sequencing and transcriptome sequencing on 81 gastric cancer tissue samples at Tianjin Medical University Cancer Institute&Hos-pital(TJMUCH)from December 2010 to December 2012.We collected clinicopathological data,compared the correlation between RYR1 mutations and expression levels,and analyzed the impact of RYR1 on the prognosis of patients with gastric cancer.Additionally,we explored the underlying molecular mechanism to study its role in promoting the development of gastric cancer by generating stable cell lines overex-pressing RYR1.Results:In TCGA gastric cancer patients,the mutation rate of RYR1 in Asian population was higher than that in others popula-tion(12.68%vs.8.13%).In gastric cancer patients from TJMUCH,RYR1 mutations ranked ninth in frequency,with a mutation rate of 33.33%.Mutations in RYR1 were negatively correlated with RYR1 expression(P=0.006 9,P<0.000 1).Patients with high RYR1 expression had significantly worse overall survival than those with low RYR1 expression(P=0.009 0,P=0.042 0).Overexpression of RYR1 promoted prolifera-tion,migration,invasion and reduced apoptosis of gastric cancer cell lines.Moreover,RYR1 overexpression was associated with decreased sensitivity to chemotherapeutic drugs in gastric cancer cells.Inhibiting RYR1-mediated calcium over-release could suppress malignant beha-viors and reverse chemoresistance.Conclusions:RYR1 had a high mutation rate in Asian gastric cancer patients and a significantly negative correlation with RYR1 mRNA levels.High RYR1 expression serves as a novel prognostic predictive marker for gastric cancer.RYR1 overex-pression promoted malignant progression of gastric cancer and chemoresistance by increasing the release of calcium ions from the endo-plasmic reticulum.Thus,RYR1 inhibition can reduce the proliferation,migration,and invasion of gastric cancer cells and reverse chemores-istance,which highlights potential combination therapies for gastric cancer.

Objective    To compare and analyze the therapeutic effects of robot-assisted lobectomy and segmentectomy for stage ⅠA non-small cell lung cancer with a diameter≤2 cm. Methods    A total of 181 patients with pathologically confirmed stage ⅠA non-small cell lung cancer (diameter≤2 cm) who underwent robot-assisted lobectomy and segmentectomy in our hospital from 2018 to 2021 were included. There were 74 males and 107 females with an average age of 57.50±10.60 years. They were divided into two groups according to the surgical procedure: a segmentectomy group (85 patients) and a lobectomy group (96 patients). Results    There was no statistically significant  difference between the two groups in terms of clinical data such as age, gender, smoking history, basic disease, pathological type, tumour diameter, operative time, postoperative 24 h drainage volume and overall complications (P>0.05). The intraoperative blood loss (33.88±16.26 mL vs. 39.27±19.48 mL, P=0.046), groups of dissected lymph nodes (4.76±1.19 vs. 5.52±1.46, P=0.000), number of dissected lymph nodes (14.81±7.23 vs. 18.06±7.70, P=0.004) and postoperative 72 h drainage volume (561.65±225.31 mL vs. 649.84±324.34 mL, P=0.037) of patients in the segmentectomy were less than those in the lobectomy group. The chest drainage time (5.49±3.92 d vs. 7.60±4.96 d, P=0.002) and postoperative hospital stay time (7.47±4.16 d vs. 9.67±5.50 d, P=0.003) were shorter than those in the lobectomy group. There was no conversion to thoracotomy or perioperative death in the two groups. The postoperative follow-up rate was 100.0% with a longest follow-up time of 48 months. The 3-year recurrence-free survival rates of the segmentectomy group and lobectomy group were 87.7% and 92.4%, respectively (P=0.465). Conclusion    The da Vinci robot-assisted lobectomy and segmentectomy are safe and feasible surgical procedures for patients with stage ⅠA non-small cell lung cancer (diameter≤2 cm), with a similar 3-year recurrence-free survival rate. The lobectomy group has more lymph nodes dissected, while the segmentectomy group is superior to the lobectomy group in terms of intraoperative blood loss, postoperative 72 h chest drainage volume, chest drainage time and postoperative hospitalization time.

Objective:To explore whether the electroacupunture stimulation (ES) at acupoint Zusanli (ST36) can inhibit the bone loss caused by Staphylococcus aureus (SA) infection and its mechanism in a model of SA osteomyelitis.Methods:Twelve male C57 BL/6 mice aged 10 to 12 weeks were randomly divided into 2 even groups ( n=6) for SA infection + ES or SA infection only. After ES at ST36 was conducted for 4 weeks in the model of SA osteomyelitis, samples were harvested from the femora and tibiae. Micro-CT reconstruction was performed to detect trabecular bone volume fraction (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), connectivity density (Conn.Dn) to analyze changes in bone mass. Leptin receptor (LEPR) staining was performed to detect osteoblasts. Tartrate resistant acid phosphatase (TRAP) staining was used to detect the changes in osteoclasts. The changes in plasma inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). Results:Micro-CT results showed that the BV/TV, Tb.N, Tb.Th, and Conn.Dn in the cancellous bone in the target areas in the SA + ES group were all higher than those in the SA group, LEPR immunofluorescence results indicated that the number of osteogenic precursor cells in the ES group was larger than that in the SA group, and serum ELISA indicated a decrease in inflammatory factors in the blood in the SA+ES group compared with the SA group. There were significant differences in the comparisons above ( P<0.05). There was no significant difference in the number of osteoclasts on the surface of trabecular bone between the 2 groups in TRAP staining. Conclusion:ES may slow down infectious bone destruction by inhibiting the inflammatory response induced by SA infection and by inducing aggregation and differentiation of mesenchymal stem cells into trabecular bone.

Objective    To compare the postoperative chylothorax outcomes of robot-assisted thoracic surgery (RATS) and video-assisted thoracoscopic surgery (VATS), analyze the risk factors for postoperative chylothorax after minimally invasive radical lung cancer resection and explore possible prevention and control measures. Methods    Between June 2012 and September 2020, 1 083 patients underwent minimally invasive pulmonary lobectomy and systematic lymph node dissection in our hospital, including 578 males and 505 females with an average age of 60.6±9.4 years. Patients were divided into two groups according to the operation methods: a RATS group (499 patients) and a VATS group (584 patients). After propensity score matching, 434 patients were included in each group (868 patients in total). Chylothorax and other perioperative indicators were compared between the two groups. Univariate and multivariate logistic regression analyses were performed to identify risk factors for postoperative chylothorax. Results    Overall, 24 patients were diagnosed with chylothorax after surgery. Compared with the VATS group, the rate of chylothorax was higher (3.9% vs. 1.6%, P=0.038), the groups and numbers of dissected lymph nodes were more (both P<0.001), and the intraoperative blood loss was significantly less (P<0.001) in the RATS group. There was no statistical difference in the postoperative hospital stay (P=0.256) or chest tube drainage time (P=0.504) between the two groups. Univariate analysis showed that gender (P=0.021), operation approach (P=0.045), smoking (P=0.001) and the groups of dissected lymph nodes (P<0.001) were significantly associated with the development of chylothorax. Multivariate analysis showed that smoking [OR=4.344, 95%CI (1.149, 16.417), P=0.030] and the groups of dissected lymph nodes [OR=1.680, 95%CI (1.221, 2.311), P=0.001] were the independent risk factors for postoperative chylothorax. Conclusion    Compared with the VATS, the rate of chylothorax after RATS is higher with more dissected lymph nodes and less blood loss. The incidence of chylothorax after minimally invasive radical lung cancer resection is higher in the patients with increased dissected lymph node groups and smoking history.