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 evaluate the therapeutic efficacy of stenting combined with medication versus medication alone in treating severe extracranial carotid artery stenosis.Methods This prospective,multicenter cohort study enrolled 337 patients with carotid artery stenosis ranging from 70%to 99%.Based on patient preference,they were divided into two groups:a stent combined with medication group(antiplatelet drugs+statins,n=169)and a medication-only group(n=168).The study assessed perioperative safety and the incidence of recurrent cerebrovascular events related to the target vessel within one year.Results The success rate of the stent combined with medication group was 100%.There was one case of hyperperfusion-related intracranial hemorrhage during the perioperative period(1/169)The patient recovered well after medical treatment.There was no statistically significant difference in perioperative complications between the two groups(P=0.318).During the 1-year follow-up,the stent combined with medication group(3/169)had a significantly lower incidence of recurrent ischemic stroke compared to the medication-only group(15/168),with a statistically significant difference(P=0.003).Univariate analysis showed that gender(χ2=4.372,P=0.037),stenosis rate≥85%(χ2=10.738,P=0.001),and non-stent treatment(χ2=7.171,P=0.007)were risk factors for recurrent stroke.However,the number of recurrent ischemic strokes in females was relatively small and after adjusting the chi-square test,the difference was not statistically significant(P=0.075).Logistic regression analysis revealed that stenosis rate≥85%(OR=115.925,95%CI:2.077-122.102,P=0.008)and non-stent treatment(OR=0.158,95%CI:0.044-0.564,P=0.004)were independent risk factors for recurrent cerebrovascular events within one year.Conclusions In patients with≥85%extracranial carotid artery stenosis,stenting combined with medication reduces recurrent ischemic cerebrovascular events within 1 year.

Objective To evaluate the therapeutic efficacy of stenting combined with medication versus medication alone in treating severe extracranial carotid artery stenosis.Methods This prospective,multicenter cohort study enrolled 337 patients with carotid artery stenosis ranging from 70%to 99%.Based on patient preference,they were divided into two groups:a stent combined with medication group(antiplatelet drugs+statins,n=169)and a medication-only group(n=168).The study assessed perioperative safety and the incidence of recurrent cerebrovascular events related to the target vessel within one year.Results The success rate of the stent combined with medication group was 100%.There was one case of hyperperfusion-related intracranial hemorrhage during the perioperative period(1/169)The patient recovered well after medical treatment.There was no statistically significant difference in perioperative complications between the two groups(P=0.318).During the 1-year follow-up,the stent combined with medication group(3/169)had a significantly lower incidence of recurrent ischemic stroke compared to the medication-only group(15/168),with a statistically significant difference(P=0.003).Univariate analysis showed that gender(χ2=4.372,P=0.037),stenosis rate≥85%(χ2=10.738,P=0.001),and non-stent treatment(χ2=7.171,P=0.007)were risk factors for recurrent stroke.However,the number of recurrent ischemic strokes in females was relatively small and after adjusting the chi-square test,the difference was not statistically significant(P=0.075).Logistic regression analysis revealed that stenosis rate≥85%(OR=115.925,95%CI:2.077-122.102,P=0.008)and non-stent treatment(OR=0.158,95%CI:0.044-0.564,P=0.004)were independent risk factors for recurrent cerebrovascular events within one year.Conclusions In patients with≥85%extracranial carotid artery stenosis,stenting combined with medication reduces recurrent ischemic cerebrovascular events within 1 year.

Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment, with the predominant clinical diagnosis of spatial working memory (SWM) deficiency, which seriously affects the physical and mental health of patients. However, the current pharmacological therapies have unsatisfactory cure rates and other problems, so non-pharmacological physical therapies have gradually received widespread attention. Recently, a novel treatment using 40 Hz light flicker stimulation (40 Hz-LFS) to rescue the cognitive function of model animals with AD has made initial progress, but the neurophysiological mechanism remains unclear. Therefore, this paper will explore the potential neural mechanisms underlying the modulation of SWM by 40 Hz-LFS based on cross-frequency coupling (CFC). Ten adult Wistar rats were first subjected to acute LFS at frequencies of 20, 40, and 60 Hz. The entrainment effect of LFS with different frequency on neural oscillations in the hippocampus (HPC) and medial prefrontal cortex (mPFC) was analyzed. The results showed that acute 40 Hz-LFS was able to develop strong entrainment and significantly modulate the oscillation power of the low-frequency gamma (lγ) rhythms. The rats were then randomly divided into experimental and control groups of 5 rats each for a long-term 40 Hz-LFS (7 d). Their SWM function was assessed by a T-maze task, and the CFC changes in the HPC-mPFC circuit were analyzed by phase-amplitude coupling (PAC). The results showed that the behavioral performance of the experimental group was improved and the PAC of θ-lγ rhythm was enhanced, and the difference was statistically significant. The results of this paper suggested that the long-term 40 Hz-LFS effectively improved SWM function in rats, which may be attributed to its enhanced communication of different rhythmic oscillations in the relevant neural circuits. It is expected that the study in this paper will build a foundation for further research on the mechanism of 40 Hz-LFS to improve cognitive function and promote its clinical application in the future.
Humans ; Adult ; Rats ; Animals ; Memory, Short-Term/physiology* ; Rats, Wistar ; Neurodegenerative Diseases ; Hippocampus ; Prefrontal Cortex

Polyhydroxyalkanoates (PHA) synthesis by activated sludge using volatile fatty acids (VFAs) in fermentation liquid of excess sludge as carbon source is a hotspot in the field of environmental biotechnology. However, there is no unified conclusion on the effects of non-VFAs, mainly dissolved organic matter (DOM), on PHA production. Thus, this critical review mainly introduces the main characteristics and common analysis methods of DOM in anaerobic fermentation liquid. The effects of DOM on PHA production are analyzed from the aspects of microbiology, metabolic regulation and sludge properties. The results of different studies showed that high concentration of DOM is bad for PHA production, but an appropriate amount of DOM is conducive to the stability of sludge properties, reducing the final PHA purification cost. Finally, suitable strategies were proposed to regulate the PHA synthesis by activated sludge with DOM for PHA production by anaerobic fermentation liquid.
Anaerobiosis ; Bioreactors ; Fatty Acids, Volatile ; Fermentation ; Polyhydroxyalkanoates ; Sewage

The withdrawal of drugs from the market due to serious adverse reactions arising from the risk of cardiac arrhythmia has been a major concern for regulatory authorities in recent years. In 2005, the International Council for the Harmonisation of Registration of Medicinal Products for Human Use (ICH) issued preclinical S7B and clinical E14 guidelines for the evaluation of arrhythmia risk. The former includes in vitro hERG experiments and in vivo animal-based QT studies, while the latter is the TQT study (thorough QT study, TQT study). The high cost and conservative results of TQT studies have led to the introduction of new alternative methods, and the E14 guidelines were revised in 2015 and introduced concentration-QT analysis (C-QTc), based on the PK/PD (pharmacokinetics/pharmacodynamics) concept, by making full use of high-quality double-negative data from the non-clinical phase (double-negative, low risk of hERG tests at high clinical exposure, and low risk of QTc prolongation in vivo QTc study) makes it possible to substitute TQT studies based on specific situations. This article reviews the history of the development of E14 and its latest developments, analyses the specific situations in which C-QT can replace TQT studies through case studies, introduces the preclinical-clinical arrhythmia risk stage assessment system and compares the drug assessment process and decision making under different versions of E14 guidelines, with a view to providing a reference for the arrhythmia risk assessment system in China.

Objective:To observe the effect of CoCl2on the cisplatin sensitivity of human ovarian cancer SKOV3cells, and to clarify the possible mechanism.Methods:The SKOV3cells were cultured in vitro and randomly divided into control group, CoCl2 group, cisplatin (DDP) group and CoCl2 combined with DDP (combination) group.The cells in CoCl2group were cultured in normal cell medium for 20hafter cultured in 200μmol·L-1 CoCl2for 4h, the cells in DDP group were cultured in normal cell medium containing 10mg·L-1 DDP for 24h, and the cells in combination group were cultured in 10mg·L-1 DDP for 20hafter cultured in 200μmol·L-1 CoCl2for 4h.The survival rates of SKOV3cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1α (HIF-1α) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method.Rhod 2-AM fluorescence probe was used to observe the levels of Ca2+in mitochondria in the cells in various groups.Western blotting method was used to observe the expression levels of cytochrome C (cyto C) , cysteinyl aspartase 3 (caspase 3) and cleaved cysteinyl aspartase 3 (cleaved caspase 3) .Muse○R apoptosis assay kit was used to detect the apoptotic rates of cells in various groups.Results:Compared with control group, the survival rate of the cells in CoCl2group had no significant change (P＞0.05) , and the survival rates of the cells in DDP and combination groups were decreased (P＜0.05) ;the survival rate in combination group was higher than that in DDP group (P＜0.05) .Compared with control group, the positive expression intensities of HIF-1αin CoCl2and combination groups were increased (P＜0.05) .Compared with control group, the positive expressions of iNOS in DDP and combination groups were increased (P＜0.05) .The Ca2+levels in the cells in DDP group and combination groups were higher than that in control group (P＜0.05) and the Ca2+level in DDP group was higher than that in combination group (P＜0.05) .Compared with control group, the expression levels of cyto C, caspase 3and cleaved caspase 3proteins in the SKOV3cells in CoCl2group had no significant changes (P＞0.05) , and the expression levels of cyto C, caspase 3and cleaved caspase 3in DDP group were increased significantly (P＜0.05) ;compared with DDP group, they were lower than those in combination group (P＜0.05) .Compared with control group, the apoptotic rate of SKOV3cells in DDP group was increased significantly (P＜0.05) ;the apoptotic rate of SKOV3cells in combination group was lower than that in DDP group (P＜0.05) .Conclusion:CoCl2can redece the mitochondrial apoptosis of human ovarian cancer SKOV3cells by inhibiting the DDP-induced enhancement of iNOS expression and decrease the sensitivity of SKOV3cells to cisplatin.

With the development of informatization in healthcare system, it is necessary to build the electronic healthcare records. This article reviews the development and meanings of using the standardize nursing terminology in electronic healthcare records, and discusses the challenges and methods of the development of standardized nursing terminology in electronic healthcare records.

Mental fatigue is a subjective fatigue state caused by long-term brain activity, which is the core of health problems among brainworkers. However, its influence on the process of brain information transmission integration is not clear. In this paper, phase amplitude coupling (PAC) between theta and gamma rhythm was used to study the electroencephalogram (EEG) data recorded before and after mental fatigue, so as to explain the effect of mental fatigue on brain information transmission mechanism. The experiment used a 4-hour professional English reading to induce brain fatigue. EEG signals of 14 male undergraduate volunteers before and after mental fatigue were recorded by Neuroscan EEG system. Phase amplitude coupling value was calculated and analyzed. test was used to compare the results between two states. The results showed that theta phase of more than 90% of the electrodes in the whole brain area jointly modulated gamma amplitude of the right central area and the right parietal area, and the coupling effect among different brain regions significantly decreased ( < 0.05) when participants had felt mental fatigue. This paper shows that phase amplitude coupling can explain the influence of mental fatigue on information transmission mechanism. It could be an important indicator for mental fatigue detection. On the other hand, the results also provide a new measure to evaluate the effect of neuromodulation in relieving mental fatigue.

Objective To investigate the diagnostic value of dynamic-extended focused assessment with sonography for trauma (D-EFAST) in patients with multiple trauma in intensive care unit (ICU). Methods A prospective clinical study was conducted. Eighty patients with multiple trauma admitted to ICU of Anhui Provincial Hospital from September 1st, 2014 to December 31st, 2016 were enrolled. Extended focused assessment with sonography for trauma (E-FAST) check was conducted at first, for those who had positive findings diagnosis was confirmed by immediately CT examination or surgical exploration. If it was negative, the patients received E-FAST every morning for 7 days (defined as D-EFAST), for those with positive findings, immediately CT or surgery was performed to clarify the diagnosis. The final clinical diagnosis was used as the "gold standard" to calculate the diagnostic accordance rate of EFAST and D-EFAST examination technique for pneumothorax, pleural effusion, spleen injury, kidney damage, liver damage, gastrointestinal injury, pericardial effusion, bladder rupture, and pancreatic injury, as well as their sensitivity, specificity, positive predictive value, negative predictive value, accuracy rate, and missed diagnosis rate, and the difference between EFAST and D-EFAST was compared. Results There were 4 patients excluded because of death and abandoning treatment, and finally 76 patients were included in the study. The total sensitivity of E-FAST examination technique for pneumothorax, pleural effusion, spleen injury, liver damage, gastrointestinal injury, pericardial effusion, and bladder rupture was 75.9% (66/87), and the specificity was 98.3% (587/597), the positive predictive value was 86.8% (66/76), and the negative predictive value was 96.5% (587/608), the accuracy rate was 95.5% (653/684), and the rate of missed diagnosis was 24.1% (21/87). The most of the delayed injury in patients with multiple trauma occurred at 2-7 days after injury with incidence of 4.8% (33/684). The diagnostic sensitivity of D-EFAST for delayed injury was 98.3% (118/120), the specificity was 99.8% (563/564), the positive predictive value was 99.2% (118/119), the negative predictive value was 99.6% (563/565), the diagnostic accuracy rate was 99.6% (681/684), and rate of missed diagnosis was 1.7% (2/120). When the final clinical diagnosis was set as the "gold standard", D-EFAST technology for the detection rate was 98.3% (118/120) for patients with multiple trauma on organ injury while the detection rate of E-FAST was 75.9% (66/87), with statistical significant difference (P < 0.01), indicating that D-EFAST was better than E-FAST in check of multiple trauma patients with organ injury. Conclusion Although the E-FAST technology can quickly diagnose the multiple trauma patients and win the rescue time for critical patients, multiple trauma patients injured after 2-7 days prone to delayed damage and are difficult to detect, and D-EFAST can be used to find delayed damage earlier, and reduce the misdiagnosis rate of multiple trauma patients.