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

Drug therapy is a common method to cure diseases and relieve symptoms.The value of patient-reported outcome(PRO)in evaluating the effect of drug therapy has been increasingly paid attention.The PRO scale is a standardized questionnaire,which can scientifically evaluate the experiences and subjective effects of drug use from a patient-centered perspective,and help patients and clinicians make more reasonable medication decisions.By reviewing and sorting out relevant global literature,this paper found that the content of the PRO scales relevant to drug therapy focused on five fields:"medication satisfaction""medication adherence""drug treatment burden""medication-related quality of life"and"adverse drug reactions".This paper described the basic information,measurement characteristics and application of common scales in recent years respectively,and summarized and analyzed the problems and enlightenment of scale development,aiming to provide theoretical reference for the selection,application and development of PRO scales.

Pharmacy services in grassroots medical institutions provide professional pharmacy services for patients. This service not only expanded the scope of primary care delivery, but also meets the residents′need for appropriate medication. At present, the majority of grassroots medical institutions in China provide outpatient pharmacy services by means of pharmacy service alliance, and foreign grassroots medical institutions mainly provide pharmaceutical outpatient services with drug treatment management as the core for patients by granting pharmacists partial prescription rights.In recent years, pharmaceutical outpatient services in grassroots medical institutions had achieved good results in improving patient clinical outcomes, enhancing medication rationality, and enhancing patient compliance. However, there were many unresolved issues, including low public awareness, shortage of pharmaceutical talents, and imperfect guarantee systems. The author suggestted that efforts should be made to increase the publicity of pharmaceutical outpatient services, improve the training mechanism for pharmacists, strengthen cooperation between pharmacists and clinical physicians, include pharmacists in the contracted service team of family doctors, and improve various guarantee systems, in order to provide reference for promoting the sustainable development of pharmaceutical outpatient services in grassroots medical institutions in China.

Objective:To explore the differences of clinical features and surgical efficacy between the elderly and the middle-aged and young patients with meningiomas in the central cortex area.Methods:Forty-three elderly patients with meningiomas in the central cortex area (≥60 years old) and 63 middle-aged and young patients with meningiomas in the central cortex area (18-59 years old), accepted surgery in Department of Neurosurgery, Guangdong 999 Brain Hospital from January 2011 to December 2020, were chosen. The differences of symptom, gross tumor volume, peritumoral edema degrees, intraoperative blood loss, tumor resection degrees, complications, and hospital stays between the 2 groups were analyzed.Results:The elderly patients had significantly higher proportion of preoperative muscle strength decline, lower proportion of preoperative headache, larger preoperative tumor volume, more severe peritumoral edema degrees before and after surgery, small volume of intraoperative blood loss, and longer hospital stays compared with the middle-aged and young patients ( P<0.05). However, no significant differences in distributions of tumor resection Simpson grades, proportion of new neurological dysfunction, incidence of postoperative complications, and proportions of reoperations and tumor recurrence were noted between the 2 groups ( P>0.05). Significant difference in distributions of peritumoral edema degrees among patients with different WHO grades was noted in the elderly group ( P<0.05), and significant difference in distribution of peritumoral edema degrees among patients with different tumor volumes was noted in the middle-aged and young group ( P<0.05). Conclusions:Although differences in clinical characteristics exist between the elderly and the middle-aged and young patients with meningiomas in the central cortex area, no significant difference in surgical efficacy is noted between the 2 groups. The peritumoral edema degree in the elderly patients is related to tumor pathological grades, while that in middle-aged and young patients is related to tumor volumes.

Objective:To evaluate the safety and tolerance of sequential thoracic radiotherapy combined with PD-1/PD-L1 inhibitors in patients with extensive-stage small cell lung cancer (ES-SCLC) after induction systemic therapy.Methods:ES-SCLC patients from a phase I trial and a real-world study were enrolled for those who received thoracic radiotherapy after induction systemic treatment (chemotherapy/chemotherapy combined with PD-1/PD-L1 inhibitors) and consolidated with PD-1/PD-L1 inhibitors. These two studies were both approved by the Ethics Committee of Chinese Academy of Medical Sciences Cancer Hospital (Clinical Trials.gov number, NCT03971214, NCT04947774).Results:Between January 2019 and March 2021, a total of 11 patients with ES-SCLC were analyzed, aged 52-73 years, with a median age of 62 years. Among them, five patients (45.5%) received induction chemotherapy and six patients (54.5%) received chemotherapy combined with PD-1/PD-L1 inhibitor, and then all received intensity-modulated thoracic radiotherapy after evaluation of systemic treatment efficacy. Two patients developed treatment-related grade G3-5 toxicity (18.2%, 1 treatment-related pneumonitis and 1 radiation esophagitis). G 1-G 2 hematologic toxicity, pneumonia, and anorexia were common mild toxicities. Only one patient (9.1%) terminated immunotherapy due to immune-related pneumonitis. During a median follow-up time of 12.5 months (range: 3.5-16.4 months), the median disease progression-free survival and overall survival was 7.4 months (95% CI: 6.9-8.0 months) and 14.6 months (95% CI: 9.0-20.2 months), respectively. Conclusions:Sequential thoracic radiotherapy followed by PD-1/PD-L1 inhibitor is safe and feasible in patients with ES-SCLC after induction therapy. Given that both thoracic radiotherapy and immunotherapy benefits the ES-SCLC in survival, this comprehensive treatment modality warrants further investigation.

OBJECTIVE：To study the improvement effects of ginsenoside Rb 3 combined with β-asarone on vascular dementia （VD）model mice and its mechanism. METHODS ：ICR mice were randomly divided into model group ，ginsenoside Rb 3 group（10 mg/kg），β-asarone group （10 mg/kg），drug combination group （ginsenoside Rb 3 10 mg/kg+β-asarone 10 mg/kg），positive control group（donepezil hydrochloride 1 mg/kg）and Akt inhibitor group （LY294002，1 mg/kg），and sham operation group was set up ， with 10 mice in each group. Except for sham operation group ，VD model was induced by four vessel occlusion method in other groups. After modeling ，sham operation group and model group were given constant volume of normal saline ，Akt inhibitor group was given relevant medicine intraperitoneally ，and other groups were given relevant medicine intragastrically ，twice a day ，for consecutive 30 d. After last administration ，the learning and memory ability of mice was detected by avoiding darkness test. The contents of 4-hydroxydecenoic acid （4-HNE），8-hydroxydeoxyguanosine （8-OHdG） and reactive oxygen species （ROS） in hippocampus was detected by ELISA. RT-PCR assay was used ， to detect the mRNA expression of Bcl- 2 and Bax inhippocampus. The protein expression of Bcl- 2 in cortex wadetected by immunofluorescence method. Western blotting deng- assay was used to detect the protein expression of Bcl- 2 and mz1@126.com Bax in hippocampus. RESULTS ： Compared with sham operation group ，the incubation period of avoiding darkness xiaoyinlanlp@126.com test in model group was shortened significantly ； and the number of errors was increased significa ntly；4-HNE，8-OHdG and ROS contents ，mRNA and protein expression of Bax were increased significantly ，and mRNA and protein expression of Bcl- 2 was decreased significantly （P＜0.01）. Compared with model group，the incubation period of avoiding darkness test was prolonged significantly in ginsenoside Rb 3 group，β-asarone group ，drug combination group and positive control group ，the number of errors was decreased significantly ；4-HNE，8-OHdG，ROS contents ， mRNA and protein expression of Bax were decreased significantly ，and mRNA and protein expression of Bcl- 2 were increased significantly，especially in drug combination group （P＜0.05 or P＜0.01）. But the incubation period of avoiding darkness test was shortened significantly in Akt inhibitor group ，and the number of errors was increased significantly ；4-HNE，8-OHdG，ROS contents，mRNA and protein expression of Bax were increased significantly ，and mRNA and protein expression of Bcl- 2 were decreased significantly （P＜0.05）. CONCLUSIONS ：Ginsenoside Rb 3 combined with β-asarone has a protective effect on VD model mice ，and the effect was better than that of each compound alone. The mechanism of which may be associated with anti-oxidative stress and anti-apoptosis of hippocampus.

Objective To evaluate the effect of an inverse planning simulated annealing (IPSA) in the treatment of cervical cancer with combined intracavitary and interstitial three-dimensional brachytherapy.Methods A total of 60 patients with locally advanced cervical cancer who received both external beam radiotherapy and combined intracavitary and interstitial brachytherapy in our hospital from October 2016 to July 2018 were enrolled.Patients were divided into four groups with 15 patients each according to the number of needles applied (1,2,3,and 4 needles,respectively).Dosimetric distributions were optimized with both Graphical optimization (GRO) and IPSA.Paired t-test was applied to compare the dosimetric differences between plans optimized with GRO and IPSA.Results The Dg0 and V100 of IPSA plans were higher than those of GRO (t=-4.742,-4.823,P＜0.05),while the conformity index (CI) and conformal index (COIN) were slightly lower than those of GRO plans (t=9.642,8.783,P＜0.05).No significant difference in the V150,V200,V300 between IPSA and GRO (P＞0.05) was observed.There was also no significant difference in the D2cm3 of bladder and rectum between IPSA and GRO (P＞0.05).The difference of Dg0 between IPSA and GRO was increased as the number of implanted needles increased,which increased from 4 cGy to 14 cGy as the number of needle increased from 1 to 4.The difference of V100 between GRO and IPSA was also increased as the number of needle increased.Conclusions In the treatment of cervical cancer with combined intracavitary and interstitial threedimensional brachytherapy,IPSA plan could improve the target coverage(D90,V100)without increasing the dose to the OARs and high dose region in the target compared with GRO.With the numbers of needles increased,the advantage of IPSA increased in terms of target coverage.

Objective:To analyze the sedative effect of dexmedetomidine in the elderly patients with intraspinal anesthesia .Meth-ods:Totally 52 elderly patients with intraspinal anesthesia were randomly divided into the observation group (26 cases) and the control group (26 cases).After anesthetized successfully , the patients in the control group were given normal saline with intravenous pum-ping,while those in the observation group were given dexmedetomidine with intravenous pumping .The levels of blood pressure , RR, SpO2and HR, the Ramsay sedation score and the adverse reactions of the two groups before anesthesia (T0),10 min after anesthesia (T1),30 min after anesthesia (T2),60 min after anesthesia (T3) and at the end of operation (T4) were compared.Results:The lev-els of RR, SpO2 and HR in the two groups were maintained within the normal range , while the levels of SBP ,HR at T2 and T3 were sig-nificantly lower than those at T 0 in the observation group , and also lower than those at the same time point in the control group ( P<0.05).The Ramsay sedation score from T2 toT4 were significantly lower than that at T0 in the observation group, and also lower than that at the same time point in the control group (P<0.05).The incidence of adverse reactions in the observation group was obviously lower than that in the control group .Conclusion: Dexmedetomidine used in the elderly patients with intraspinal anesthesia can keep hemodynamic stability and exhibits adequate sedation with significantly reduced adverse reactions , which is worthy of clinical promo-tion.

Objective The method for detecting expression of human CDK14 gene with Real-time quantitative PCR was developed.Methods To establish a method for detecting expression of human CDK14 gene with Real-time quantitative PCR by designing and synthesis of the primers of CDK14 target gene andβ-Actin reference gene and extracting total RNA from different lung cancer cell lines.Then the specificity,detection range and repeatability of this method were evaluated.At last,the expression level of CDK14 gene in different cell lines,which were with or without siRNA interference,were carried out by using this method.Results The method for detecting expression of human CDK14 gene with Real-time quantitative PCR,which had good specificity,good repeatability (CV=7.3 ％) and wide detection range (Ct value range of CDK14 and β-Actin amplification curve were 22.47～32.96 and 15.14～ 27.55 respectively,r2 =0.9844),was developed and it was verified by electrophoresis analysis,melting curve,PCR product sequencing.And CDK14 gene expression level,which was detected by this method,increased in HCC827 D5,H1650 and number 1 siRNA segment was effective interference segment.Conclusion The method for detecting expression of human CDK14 gene with Real-time quantitauve PCR was established successfully.

Objective To evaluate the procedures for patient-specific quality assurance ( QA ) measurements using modulated spot scanning beam for proton and carbon ion treatment. Methods Records of 1734 beam measurements were analyzed by using a passing criteria, namely, dose difference between the measured and calculated doses≤3% or distance-to-agreement ( DTA) between them≤3 mm, and meanwhile mean dose deviation in all chambers ≤3% and at least 22 of 24 chamber array having passed this criteria. The results of measurement were categoried by water phantom, treatment room, measurement depth in chamber array, etc. Results Stratification of result showed some correlation between measurement parameters and passing rates. The total passing rates were 97. 7% and 91. 9% for proton and carbon ion beams, respectively. The passing rates were high at all measurement depths for proton beam, but decreased for carbon ion beam with increased depth. Chambers were mostly stable, leading to no significant difference in passing rate between different water phantoms and between different treatment rooms. Conclusions A good agreement was shown between the doses measured by water phantoms with those by the treatment planning system ( TPS) . When the chamber position was deeper than 150 mm in carbon ion measurements, a lower passing rate was observed. This could be caused by uncertainty of ion chamber array setup ( lateral and in depth) in highly modulated beams or by incorrect modelling of scattering by the TPS. These deviations need the further investigation.