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 assess the clinical value of multimodal ultrasound score for assessment of endometrial receptivity among patients with polycystic ovary syndrome (PCOS), and to provide guidance for improving pregnancy outcomes among PCOS patients. Methods A total of 48 PCOS patients admitted to Jiangning Hospital Affiliated to Nanjing Medical University between January and December 2023 were enrolled as the case group, while 50 healthy women of childbearing age received ovulation monitoring at the same hospital during the same period served as the control group. Subjects received two-dimensional grayscale ultrasound during the implantation window (19 to 23 days of the menstrual cycle) for measurement of endometrial thickness, Gonen classification, and endometrial peristalsis. Two-dimensional color Doppler ultrasound was used for assessment of endometrial blood flow and three-dimensional ultrasound was used for assessment of endometrial volume and vascularization flow index (VFI). The endometrium multimodal ultrasound scores were estimated, and various parameters were compared between the two groups. The diagnostic performance of these parameters for PCOS was evaluated with receiver operating characteristic (ROC) curves. Results The age of subjects in the case group ranged from 20 to 38 years, with a mean age of (28.20 ± 2.82) years, and their body mass index (BMI) ranged from 21.23 to 29.11 kg/m², with a mean BMI of (26.25 ± 1.60) kg/m². The age of subjects in the control group ranged from 22 to 38 years, with a mean age of (28.10 ± 1.99) years, and their BMI ranged from 21.33 to 29.03 kg/m², with a mean BMI of (26.10 ± 1.78) kg/m². There were no significant differences between the case and control groups in terms of mean age, BMI, estradiol, and testosterone (t = 0.218, 0.422, 0.010, and 0.221; all P > 0.05). The endometrial thickness, endometrial volume, and VFI were significantly higher in the control group than in the case group (t = 4.838, 4.978, and 7.115; all P < 0.05). There were significant differences between the two groups in terms of endometrial classification, endometrial peristalsis pattern, and endometrial and sub-endometrial blood flow (Z = −4.136, −4.048, and −3.884; all P < 0.05). The scores of endometrial classification, endometrial peristalsis, endometrial and sub-endometrial blood flow, endometrial volume, VFI, and multimodal ultrasound were significantly lower in the case group than in the control group (t = 4.539, 4.449, 4.205, 3.209, 5.206, and 4.495; all P < 0.05). No significant difference was detected in the endometrial thickness score between the two groups (t = -0.149, P = 0.882). The areas under the ROC curves for endometrial thickness, endometrial volume, VFI, and multimodal ultrasound scores in diagnosis of PCOS were 0.753, 0.747, 0.809, and 0.858, respectively. Conclusion Multimodal ultrasound score provides a comprehensive assessment of the endometrium, and is effective in the assessment of endometrial receptivity, which may provide a reference for guiding pregnancy planning in PCOS patients.

Objective:To analyze the echocardiographic and genetic characteristics of fetuses with common arterial trunk（CAT）.Methods:A retrospective analysis was conducted on 77 480 fetal echocardiograms examined at the Maternal-Fetal Medicine center in Fetal Heart Disease of Beijing Anzhen Hospital from November 2010 to November 2024.Among them，106 fetuses were initially diagnosed with CAT，and 95 cases were ultimately confirmed（0.1%，95/77 480）. The echocardiographic and genetic features of CAT fetuses were analyzed. According to the modified Van Praagh classification，CAT was divided into types A1-A4［with ventricular septal defect（VSD）］and B1-B4（without VSD）based on the origin of the pulmonary artery branches and the presence or absence of a VSD. Additionally，CAT was categorized into isolated and complex types based on the presence of associated intracardiac or extracardiac anomalies.Results:① Among the 95 confirmed CAT fetuses，type A accounted for 90.5%（86/95），and type B accounted for 9.5%（9/95）. All 9 type B CAT fetuses exhibited no overriding of the arterial trunk ， with 8 cases showing left ventricular hypoplasia accompanied by mitral atresia or absence.② Of the 95 CAT fetuses，14 were isolated（14.7%，14/95） ， and 81 were complex（85.3%，81/95）.The main associated intracardiac anomalies included：single ventricle（22 cases），complete atrioventricular septal defect（12 cases），anomalous pulmonary venous drainage（10 cases），right aortic arch with mirror-image branching（16 cases），and persistent left superior vena cava（14 cases）. ③ Genetic testing was performed in 31 fetuses，with 18 showing positive results，primarily 22q11.21 deletion syndrome（29.0%，9/31）. Conclusions:Apart from VSD，the most common intracardiac anomaly associated with CAT fetuses is single ventricle. Type B CAT without trunk overriding is often associated with left ventricular hypoplasia and mitral atresia or absence. The most frequent genetic abnormality in CAT fetuses is 22q11.21 deletion syndrome. Prenatal echocardiography should clarify the CAT subtype and associated anomalies，and genetic testing is strongly recommended for perinatal counseling and prognostic evaluation.

Objective:To compare the anterior long-segment fixation, posterior long-segment fixation, and combined anterior-posterior fixation in the treatment of ankylosing spondylitis with cervical fracture (ASCF).Methods:A retrospective study was conducted to analyze the 153 patients with ASCF who had been treated at Department of Orthopaedics, Peking University Third Hospital and Department of Orthopedics, Luoche Central Hospital between January 2014 and December 2023. The cohort included 86 males and 67 females, with an age of (41.6±11.5) years, a disease duration of (10.0±3.9) years, and an interval from injury to surgery of (3.3±1.4) d. By Frankel's classification for preoperative nerve injury, 57 cases were grade B, 51 grade C, and 45 grade D. Based on the surgical approaches, the patients were divided into 3 groups: an anterior group ( n=63) undergoing the anterior cervical long-segment fixation, a posterior group ( n=51) undergoing the posterior cervical long-segment fixation, and a combination group ( n=39) undergoing combined anterior-posterior cervical fixation. Surgical time, intraoperative blood loss, fracture healing, complications, and changes in Frankel grading for spinal cord injury were compared among the 3 groups. Results:There was no statistically significant difference in the preoperative general data among the 3 groups, indicating comparability ( P > 0.05). All patients were followed up for (33.5±12.0) months after surgery. In the anterior group, the surgical time [(103.0±16.8) min] was significantly shorter than that in the posterior group [(148.4±17.7) min] and that in the combination group [(228.5±23.9) min], the intraoperative blood loss [(92.8±27.8) mL] was significantly less than that in the posterior group [(477.5±109.5) mL] and that in the combination group [(769.5±136.9) mL], and the incidence of complications [9.5% (6/63)] was significantly lower than that in the posterior group [41.2% (21/51)] and that in the combination group [53.8% (21/39)] (all P<0.05). There was no statistically significant difference in the fracture healing time among the 3 groups ( P=0.111). At the last follow-up, X-ray and CT scans showed no loosening or breakage of internal fixation in all the 3 groups. The Frankel grading at the last follow-up: 12 cases of grade C, 15 cases of grade D, and 36 cases of grade E in the anterior group; 3 cases of grade B, 12 cases of grade C, 12 cases of grade D, and 24 cases of grade E in the posterior group; 6 cases of grade C, 12 cases of grade D, and 21 cases of grade E in the combination group. At the last follow-up, all patients showed a significant improvement compared to their Frankel grades before surgery ( P<0.001), but there was no statistically significant difference between the 3 groups ( H=2.238, P=0.327). Conclusions:In the treatment of ASCF, anterior long-segment fixation is advantageous over posterior long-segment fixation and combined anteri-or-posterior fixation due to its shorter surgical time, reduced intraoperative blood loss, and a lower complication incidence. All the 3 surgical approaches demonstrate comparable outcomes in terms of fracture healing time, radiographic stability, and final neurological recovery.

Objective To investigate the regulatory mechanism of the central nervous system in patients with refractory overactive bladder (rOAB) using diffusion tensor imaging (DTI) and graph theory analysis. Methods A total of 43 rOAB patients (rOAB group) and 46 matched healthy controls (HC group) were recruited during May and Nov.2024. All participants were scanned with DTI, and surveyed with the overactive bladder symptom score (OABSS), and overactive bladder questionnaire (OAB-q). Their age, gender, height, weight, and educational years were collected.DTI plus graph theory analysis was employed to explore the alterations in global and local topological properties of the brain structural network in rOAB patients. Brain regions showing significant group differences in structural metrics [specifically, the right paracentral lobule (PCL.R) ]were further used as seed points for functional connectivity (FC) analysis. Correlations between the nodal clustering coefficient (NCp) of the identified region, FC strength, OABSS, and OAB-q score were investigated. Results The OABSS [8 (6,10) vs.0 (0,1) ]and OAB-q [71 (53,80) vs.20 (19,24) ]were higher in the rOAB group than the HC group (P<0.001). Graph theory analysis revealed no statistically significant differences in global network metrics between the two groups (P>0.05). However, the NCp was significantly higher in the PCL.R of rOAB group compared to HC group (P<0.05, FDR-corrected).FC analysis using the PCL.R as a seed region demonstrated significantly reduced FC value in the left cerebellar crus Ⅱ (Cerebelum_Crus2_L) of the rOAB group (P<0.05, FDR-corrected). Partial correlation analysis showed that the NCp of PCL.R was positively correlated with both OABSS (r=0.255, P=0.018) and OAB-q score (r=0.257, P=0.017). Conversely, the FC of Cerebelum_Crus2_L was significantly negatively correlated with OABSS (r=-0.545, P<0.001) and OAB-q score (r=-0.535, P<0.001). Conclusion Patients with rOAB exhibit distinct brain structural network alterations compared to healthy individuals, primarily manifestation in the NCp value of PCL.R increased, and the FC intensity of Cerebelum_Crus2_L is significantly weakened. These alterations in the topological properties of the structural network may be implicated in the pathogenesis of rOAB.

Objective:To investigate the effects of percutaneous nerve electrical stimulation (TENS) combined with exercise therapy on lower limb microcirculation, lower limb muscle strength, and motor function in patients with knee osteoarthritis (KOA).Methods:Sixty patients with KOA who received treatment at Zhejiang Rongjun Hospital from January 2022 to December 2023 were included in this study. The patients were randomly divided into a control group ( n = 30) and an observation group ( n = 30) using the random number table method. The control group received exercise therapy, while the observation group received TENS and exercise therapy. Both groups were treated for 2 weeks. The efficacy was compared between the two groups. Additionally, pain levels, lower limb microcirculation, lower limb muscle strength, and motor function were compared between the two groups both before and after treatment. Results:The total effective rate in the observation group was significantly higher than that in the control group [93.33% (28/30) vs. 73.33% (22/30), χ2 = 4.32, P < 0.05]. After treatment, Visual Analog Scale score and microcirculatory blood perfusion volume in the observation group were (2.03 ± 0.41) and (7.98 ± 1.24) mL/min, respectively, both of which were significantly lower than those in the control group [(3.78 ± 0.59), (9.23 ± 1.36) mL/min, t = 13.34, 3.72, both P < 0.05]. The proportion of patients achieving Lovett grades of 4-5 in the observation group was significantly higher than that in the control group [83.33% (25/30) vs. 60.00% (18/30), Z = 1.99, P < 0.05]. After treatment, the Hospital for Special Surgery knee score in the observation group was significantly higher than that in the control group [(78.98 ± 5.65) vs. (67.87 ± 6.26), t = 7.21, P < 0.05]. Conclusions:TENS combined with exercise therapy has a good efficacy in the treatment of KOA. The combined therapy can improve lower limb microcirculation, lower limb muscle strength, and motor function.

Objective:To investigate the effects of percutaneous nerve electrical stimulation (TENS) combined with exercise therapy on lower limb microcirculation, lower limb muscle strength, and motor function in patients with knee osteoarthritis (KOA).Methods:Sixty patients with KOA who received treatment at Zhejiang Rongjun Hospital from January 2022 to December 2023 were included in this study. The patients were randomly divided into a control group ( n = 30) and an observation group ( n = 30) using the random number table method. The control group received exercise therapy, while the observation group received TENS and exercise therapy. Both groups were treated for 2 weeks. The efficacy was compared between the two groups. Additionally, pain levels, lower limb microcirculation, lower limb muscle strength, and motor function were compared between the two groups both before and after treatment. Results:The total effective rate in the observation group was significantly higher than that in the control group [93.33% (28/30) vs. 73.33% (22/30), χ2 = 4.32, P < 0.05]. After treatment, Visual Analog Scale score and microcirculatory blood perfusion volume in the observation group were (2.03 ± 0.41) and (7.98 ± 1.24) mL/min, respectively, both of which were significantly lower than those in the control group [(3.78 ± 0.59), (9.23 ± 1.36) mL/min, t = 13.34, 3.72, both P < 0.05]. The proportion of patients achieving Lovett grades of 4-5 in the observation group was significantly higher than that in the control group [83.33% (25/30) vs. 60.00% (18/30), Z = 1.99, P < 0.05]. After treatment, the Hospital for Special Surgery knee score in the observation group was significantly higher than that in the control group [(78.98 ± 5.65) vs. (67.87 ± 6.26), t = 7.21, P < 0.05]. Conclusions:TENS combined with exercise therapy has a good efficacy in the treatment of KOA. The combined therapy can improve lower limb microcirculation, lower limb muscle strength, and motor function.

Objective:To analyze the echocardiographic and genetic characteristics of fetuses with common arterial trunk（CAT）.Methods:A retrospective analysis was conducted on 77 480 fetal echocardiograms examined at the Maternal-Fetal Medicine center in Fetal Heart Disease of Beijing Anzhen Hospital from November 2010 to November 2024.Among them，106 fetuses were initially diagnosed with CAT，and 95 cases were ultimately confirmed（0.1%，95/77 480）. The echocardiographic and genetic features of CAT fetuses were analyzed. According to the modified Van Praagh classification，CAT was divided into types A1-A4［with ventricular septal defect（VSD）］and B1-B4（without VSD）based on the origin of the pulmonary artery branches and the presence or absence of a VSD. Additionally，CAT was categorized into isolated and complex types based on the presence of associated intracardiac or extracardiac anomalies.Results:① Among the 95 confirmed CAT fetuses，type A accounted for 90.5%（86/95），and type B accounted for 9.5%（9/95）. All 9 type B CAT fetuses exhibited no overriding of the arterial trunk ， with 8 cases showing left ventricular hypoplasia accompanied by mitral atresia or absence.② Of the 95 CAT fetuses，14 were isolated（14.7%，14/95） ， and 81 were complex（85.3%，81/95）.The main associated intracardiac anomalies included：single ventricle（22 cases），complete atrioventricular septal defect（12 cases），anomalous pulmonary venous drainage（10 cases），right aortic arch with mirror-image branching（16 cases），and persistent left superior vena cava（14 cases）. ③ Genetic testing was performed in 31 fetuses，with 18 showing positive results，primarily 22q11.21 deletion syndrome（29.0%，9/31）. Conclusions:Apart from VSD，the most common intracardiac anomaly associated with CAT fetuses is single ventricle. Type B CAT without trunk overriding is often associated with left ventricular hypoplasia and mitral atresia or absence. The most frequent genetic abnormality in CAT fetuses is 22q11.21 deletion syndrome. Prenatal echocardiography should clarify the CAT subtype and associated anomalies，and genetic testing is strongly recommended for perinatal counseling and prognostic evaluation.

Objective:To compare the anterior long-segment fixation, posterior long-segment fixation, and combined anterior-posterior fixation in the treatment of ankylosing spondylitis with cervical fracture (ASCF).Methods:A retrospective study was conducted to analyze the 153 patients with ASCF who had been treated at Department of Orthopaedics, Peking University Third Hospital and Department of Orthopedics, Luoche Central Hospital between January 2014 and December 2023. The cohort included 86 males and 67 females, with an age of (41.6±11.5) years, a disease duration of (10.0±3.9) years, and an interval from injury to surgery of (3.3±1.4) d. By Frankel's classification for preoperative nerve injury, 57 cases were grade B, 51 grade C, and 45 grade D. Based on the surgical approaches, the patients were divided into 3 groups: an anterior group ( n=63) undergoing the anterior cervical long-segment fixation, a posterior group ( n=51) undergoing the posterior cervical long-segment fixation, and a combination group ( n=39) undergoing combined anterior-posterior cervical fixation. Surgical time, intraoperative blood loss, fracture healing, complications, and changes in Frankel grading for spinal cord injury were compared among the 3 groups. Results:There was no statistically significant difference in the preoperative general data among the 3 groups, indicating comparability ( P > 0.05). All patients were followed up for (33.5±12.0) months after surgery. In the anterior group, the surgical time [(103.0±16.8) min] was significantly shorter than that in the posterior group [(148.4±17.7) min] and that in the combination group [(228.5±23.9) min], the intraoperative blood loss [(92.8±27.8) mL] was significantly less than that in the posterior group [(477.5±109.5) mL] and that in the combination group [(769.5±136.9) mL], and the incidence of complications [9.5% (6/63)] was significantly lower than that in the posterior group [41.2% (21/51)] and that in the combination group [53.8% (21/39)] (all P<0.05). There was no statistically significant difference in the fracture healing time among the 3 groups ( P=0.111). At the last follow-up, X-ray and CT scans showed no loosening or breakage of internal fixation in all the 3 groups. The Frankel grading at the last follow-up: 12 cases of grade C, 15 cases of grade D, and 36 cases of grade E in the anterior group; 3 cases of grade B, 12 cases of grade C, 12 cases of grade D, and 24 cases of grade E in the posterior group; 6 cases of grade C, 12 cases of grade D, and 21 cases of grade E in the combination group. At the last follow-up, all patients showed a significant improvement compared to their Frankel grades before surgery ( P<0.001), but there was no statistically significant difference between the 3 groups ( H=2.238, P=0.327). Conclusions:In the treatment of ASCF, anterior long-segment fixation is advantageous over posterior long-segment fixation and combined anteri-or-posterior fixation due to its shorter surgical time, reduced intraoperative blood loss, and a lower complication incidence. All the 3 surgical approaches demonstrate comparable outcomes in terms of fracture healing time, radiographic stability, and final neurological recovery.

Objective:To clarify the transitional components in the blood of compound Banlangen Granules; To explore the mechanism of drugs in the treatment of epidemic encephalitis B, hepatitis and parotitis.Methods:The transitional components in blood of compound Banlangen Granules were analyzed by ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS/MS). The regulatory targets and pathways of compound Banlangen Granules in the treatment of epidemic encephalitis B, hepatitis and parotitis were analyzed based on UPLC-MS/MS and network pharmacology.Results:A total of 9 blood components were identified, of which 8 were prototype components, including sucrose, o-aminobenzoic acid, uridine, adenosine, guanosine, indole-3-acetonitrile-2 murine-S-β-D-glucopyranoside and salicylic acid. Through network pharmacological analysis, it was concluded that compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.Conclusion:The 9 blood components of compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.