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 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 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.

Objective To study the influence factors on detection coagulation factors Ⅸ activity in human prothrombin complex concentrates(PCC).MethodsUsing Chinese Pharmacopoeia(2010)as reference,factor Ⅸ deficient plasma from different manufacturers,different types of instruments,different methods of heparin neutralization,different sample pretreatment methods were used to determine the activity of coagulation factor Ⅸ in PCC.The influence on the results of coagulation factor Ⅸ was analyzed.ResultsThere was a significant difference between factor Ⅸ deficient plasma from different manufacturers for the activity of coagulation factors Ⅸ(P<0.05).There was no significant difference between the results of coagulation factor Ⅸ measured by different types of Automatic Coagulation Analyzer.When the dilution ratio was more than 60 times,the neutralization of heparin had little effect on the coagulation factor Ⅸ activity.When the dilution ratio is less than 60 times,the detection results of coagulation factor Ⅸ by neutralization of heparin are higher than no neutralization of heparin,and the difference is significant(P<0.05).Samples for pre-temperature or not and whether dissolution for 15minutes or not,the coagulation factor Ⅸ activity showed no significant difference.Conclusion The coagulation factor Ⅸ activity in PCC was affected by factor Ⅸ deficient plasma from different manufacturers,different dilution times of heparin neutralization method and sample pre-treatment methods.It must be paid attention to in the detection process.Strengthen the quality control of the factor deficient plasma and the standardization of operation process are necessary.The External Quality Assessment for the detection of coagulation factors in PCC products should be edtablished.

OBJECTIVETo study the analgesic effects and mechanism of Yuanhuzhentong capsule.

METHODThe rats trigeminal nerve headache model, the migraine model caused by nitroglycerim, and the mice hot plate test. The rat formalin test were used to evaluate the analgesic effects. The effects on mice automative activities, the content of monoamine neurotansmitters in rats trigeminal nerve headache model and the hemorrheology in model of blood stasis were investigated to analyze the analgesic effects.

RESULTYuanhuzhentong capsule can prolong the latency of the rats trigeminal nerve headache model and reduce the durante dolors. It can relieve the reaction caused by nitroglycerim in different times and increase the threshold in mice hot plate test. It can decrease obviously the response time of the chronicity pain model rat caused by formalin in second phase and also decrease the counts of automative activities in 5 min. It can increase the content of 5-HT and decrease blood viscosity in blood stasis model rats.

CONCLUSIONThe results indicated that Yuanhuzhentong capsule has obvious analgesic effects, and the mechanism concernes with the sedation, adjusting the secretion of NT in brain and improving the blood circulation.

Analgesics ; administration & dosage ; Animals ; Capsules ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Headache ; drug therapy ; physiopathology ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Pain ; drug therapy ; physiopathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley