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

Purpose To investigate the clinicopathological features,molecular genetics and prognosis of high grade B cell lymphoma with concurrent MYC rearrangement and 11q aberra-tions(HGBCL-MYC-11q).MethodsThree cases of HGBCL-MYC-11q were reviewed and analyzed using hematoxylin-eosin staining,immunohistochemistry,EBER in situ hybridization and fluorescence in situ hybridization.Clinical data were collected with follow-up.Results All three patients were male,age was 10,61,and 74 years,respectively.All patients had Ann Arbor stage Ⅳ disease.All three cases were biopsies occurring in the nasopharynx,upper pharynx and ileocecus,respectively.Three cases were morphologically similar to diffuse infiltrative growth of tumor cells,moderate or moderately large cells,round to slightly irregular nuclei and easily visible mitotic figures.Focal necrosis was noted in one case.One case exhibited the distinct"starry sky"pattern.All cases expressed CD20,BCL6 and MUM1 and high Ki67 index,two cases expressed CD10 and two cases ex-pressed BCL2.CD3,CD30 and TDT were all negative.EBER in situ hybridization was all negative.FISH analyses using C-MYC break-apart probes were all positive and all cases had 11q aberrations.One case only had the 11q23.3 amplification;and one case only had the 11q24.3 loss.After a follow-up for 1-18 months,one patient died and two patients survived with disease.ConclusionHGBCL-MYC-11q is rare,morphologically similar to BL/HGBCL,with MYC rearrangement and 11q abnormali-ties.We should enhance awareness of the disease and improve more accurate diagnosis and differential diagnosis of the disease.

Flubromazolam(Flub)is a novel psychoactive substance of benzodiazepines and the mechanism underlying its addiction still remains elusive.This study investigated the reward effect of Flub using conditioned place preference(CPP)mouse model.The neuronal activity was evaluated by c-Fos expression,and the neural circuit was tracked by virus tracing.This study also investigated the regulatory effect of neural circuits on Flub-induced reward effects through chemogenetic approach.The results showed that,at the dose of 3 mg/kg,Flub significantly increased CPP score and c-Fos expression in dopaminergic(DA)neurons of ventral tegmental area(VTA).Inhibition of VTA dopaminergic neuron activity dramatically decreased Flub-induced CPP score.Virus tracing verified GABAergic neuronal projection of medial rostrum tegmental nucleus(RMTg)to VTA dopaminergic neurons.Activation of RMTgGABA→VTADA circuit or blockade of benzodiazepine receptors(BZR)in RMTg significantly decreased Flub-induced CPP score.These results indicate that Flub produced reward effect via BZR-mediated RMTgGABA→VTADA circuit.

Diabetic peripheral neuropathy（DPN） is a neurodegenerative disease of diabetes mellitus involving peripheral nervous system damage， which is characterized by axonal degenerative necrosis， Schwann cell apoptosis and demyelination of nerve myelin sheath as the main pathological features， this disease is highly prevalent and is a major cause of disability in diabetic patients. Currently， the pathogenesis of DPN may be related to oxidative stress， inflammatory response， metabolic abnormality， and microcirculation disorder. The treatment of DPN in modern medicine mainly starts from controlling blood glucose， nourishing nerves and improving microcirculation， which can only alleviate the clinical symptoms of patients， and it is difficult to fundamentally improve the pathological damage of peripheral nerves. Mitochondrial quality control refers to the physiological mechanisms that can maintain the morphology and functional homeostasis of mitochondria， including mitochondrial biogenesis， mitochondrial dynamics， mitochondrial oxidative stress and mitochondrial autophagy， and abnormal changes of which may cause damage to peripheral nerves. After reviewing the literature， it was found that traditional Chinese medicine（TCM） can improve the low level of mitochondrial biogenesis in DPN， maintain the balance of mitochondrial dynamics， inhibit mitochondrial oxidative stress and mitochondrial autophagy， and delay apoptosis of Schwann cells and neural axon damage， which has obvious effects on the treatment of DPN. With the deepening of research， mitochondrial quality control may become one of the potential targets for the research of new anti-DPN drugs， therefore， this paper summarized the research progress of TCM in treating DPN based on four aspects of mitochondrial quality control， with the aim of providing a theoretical research basis for the discovery of new drugs.

Diabetic peripheral neuropathy （DPN） is characterized by insidious onset， easy misdiagnosis， and progression to severe consequences such as diabetic foot ulcers， gangrene， and amputation. The main pathological features of DPN are nerve cell injuries， such as axonal degeneration and necrosis， segmental demyelination of nerve fibers， and apoptosis of Schwann cells. The phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway is a classical pathway that communicates intracellular and extracellular information and regulates biological activities such as cell proliferation， differentiation， apoptosis， autophagy， and migration. It widely affects various cells related to DPN. In recent years， numerous studies have found that the sustained high glucose environment causes abnormalities in the PI3K/Akt signaling pathway. This， in turn， accelerates the occurrence and development of DPN by participating in the pathogenesis of DPN， such as glucose and lipid metabolism， oxidative stress， inflammation， autophagy， apoptosis， and angiogenesis. Therefore， regulating the PI3K/Akt signaling pathway is crucial for the treatment of DPN. Currently， there is a lack of effective measures to slow down or reverse DPN in clinical practice. Traditional Chinese medicine （TCM） has unique advantages in preventing and treating DPN with multiple targets， effects， and components. A large number of animal and clinical studies of TCM treatment of DPN have shown that the PI3K/Akt signaling pathway is an important target for TCM treatment of DPN. Regulating the PI3K/Akt signaling pathway can promote myelin sheath repair and regeneration， delay the process of nerve cell death， and play a role in preventing and treating DPN. However， there is currently no systematic review and summary of this field in China and abroad. Therefore， this article summarized the regulation of the PI3K/Akt signaling pathway and its role in the pathogenesis of DPN， as well as the intervention of effective components of single Chinese medicine or compounds on the PI3K/Akt signaling pathway. This study is expected to provide a reference for the clinical diagnosis and treatment of DPN with TCM， basic research， and drug development.

Objective Exploring the effect of Tong luo tang tai(TLTT)on diabetic peripheral neuropathy(DPN)in GK rats with Wnt/β-The influence of the catenin signaling pathway.Methods Fifty GK rats were randomly divided into model group,TLTT high,medium,and low dose groups,and Western medicine group,with 10 rats in each group.Another 10 wistar rats were selected as the normal group.Except for the normal group,all other groups were fed with high fat to prepare DPN rat models.After 15 weeks,the DPN model was successfully prepared,and the rats in each group were treated by gavage.The high,medium,and low dose groups of TLTT were given traditional Chinese medicine TLTT 28 g·kg-1,14 g·kg-1,and 7 g·kg-1,respectively.The western medicine group was given metformin 100 mg·kg-1 and mecobalamin 0.2 mg·kg-1 by gavage.Rats in each group were administered once a day for 8 consecutive weeks.The general state,fasting blood sugar(FBS),thermal contraction latency(TWL),motor nerve conduction velocity(MNCV),and pathological changes in the sciatic nerve tissue were observed under transmission electron microscopy(Real time PCR)Western blot detection of wingless MMTV integration site family member 3A(Wnt3a)β Catenin(β-Catenin,Glycogen Synthesis Kinase-3β Glycogen synthesis kinase-3β,GSK-3β)MRNA and protein expression levels of antagonists(WNT inhibitor factor-1,Wif-1)on the Wnt signaling pathway.Results Compared with the normal group,the model group showed poorer general condition and significant pathological ultrastructural changes in the sciatic nerve.Its FBS level increased(P<0.01),TWL level decreased(P<0.01),and MNCV significantly slowed down(P<0.01).The model group had Wnt3a β-Catenin,GSK-3β MRNA and protein expression levels decreased(P<0.05),while Wif-1 mRNA and protein expression levels increased(P<0.01).After drug intervention,compared with the model group,the general condition and pathological ultrastructure of the sciatic nerve were improved in the TLTT high,medium,low,dose,and Western medicine groups,with a decrease in FBS levels(P<0.01)and an increase in TWL levels(P<0.05).The MNCV of each TLTT dose group and Western medicine group was significantly improved(P<0.01).The Wnt3amRNA of the TLTT high-dose group and Western medicine group was significantly increased(P<0.05),while the Wif-1mRNA of the TLTT high-dose group and Western medicine group was significantly reduced(P<0.05),There was a significant increase in Wnt3 protein in the high-dose and Western medicine groups of TLTT(P<0.01),as well as in the high-dose,medium,and low-dose TLTT and western medicine groups β-Catenin protein significantly increased(P<0.01,P<0.05),with high,medium,and low doses of TLTT and Western medicine group GSK-3β The protein significantly increased(P<0.01,P<0.05),while the Wif-1 protein significantly decreased(P<0.01,P<0.05)in the high and medium dose TTLTT and western medicine groups.Conclusion Tongluo Tangtai can alleviate sciatic nerve injury in DPN to a certain extent,and its mechanism may be related to the activation of Wnt/β,the catenin signaling pathway is involved.

Background::The potential impact of β cell function and insulin sensitivity on adverse pregnancy outcomes in women with gestational diabetes mellitus (GDM) remains uncertain. We aimed to investigate the association between β cell dysfunction, insulin resistance, and the composite adverse pregnancy outcomes.Methods::This observational study included 482 women diagnosed with GDM during pregnancy. Quantitative metrics on β cell function and insulin sensitivity during pregnancy were calculated using traditional equations. The association of β cell dysfunction and insulin resistance with the risk of the composite adverse pregnancy outcomes was investigated using multivariable-adjusted logistic regression models.Results::Multivariable-adjusted odds ratios (ORs) of adverse pregnancy outcomes across quartiles of homeostatic model assessment for insulin resistance (HOMA-IR) were 1.00, 0.95, 1.34, and 2.25, respectively ( P for trend = 0.011). When HOMA-IR was considered as a continuous variable, the multivariable-adjusted OR of adverse pregnancy outcomes was 1.34 (95% confidence interval 1.16-1.56) for each 1-unit increase in HOMA-IR. Multivariable-adjusted ORs of adverse pregnancy outcomes across quartiles of homeostatic model assessment for β cell function (HOMA-β) were 1.00, 0.51, 0.60, and 0.53, respectively ( P for trend = 0.068). When HOMA-β was considered as a continuous variable, the multivariable-adjusted OR of adverse pregnancy outcomes was 0.57 (95% CI 0.24-0.90) for each 1-unit increase in HOMA-β. However, other quantitative metrics were not associated with the composite adverse pregnancy outcomes. Conclusions::We demonstrated a significant association of β cell function and insulin sensitivity with the risk of adverse pregnancy outcomes. We have provided additional evidence on the early identification of adverse pregnancy outcomes besides the glycemic values.

Objective:To evaluate the relationship between menopausal symptoms and everyday cognition in perimenopausal and postmenopausal women.Methods:A cross-sectional study was used to analyze the clinical data of 314 women who first visited the menopausal clinic of Department of Gynecology and Obstetrics of Shanghai Jiao Tong University Affiliated Sixth People's Hospital from October 2018 to October 2020. The modified Kupperman menopausal index (KMI) scale was used to assess the prevalence and severity of menopausal symptoms, and a short version of everyday cognition (ECog) scale was used to evaluate everyday cognition. Multiple linear regression was used to analyze the relationship between menopausal symptoms and everyday cognition in perimenopausal and postmenopausal women.Results:The ECog score of 314 patients was 1.50±0.49. The score of KMI was 12.67±7.88, 74.8% (235/314) of the patients had menopausal symptoms, and 34.7% (109/314) had moderate to severe menopausal symptoms. Compared with asymptomatic patients (ECog scores: 1.27±0.29), symptomatic patients had higher ECog scores (mild symptoms group: 1.48±0.47, P=0.005; moderate to severe symptoms group: 1.69±0.55, P<0.001) . Correlation analysis showed that the five symptoms of KMI (hot flashes/sweating, insomnia, irritability, melancholia and fatigue) were associated with the decline of everyday cognition ( P<0.001). Multiple linear regression analysis showed that the symptoms of hot flashes/sweating [ β=0.060, 95% confidence interval ( CI) =0.002-0.117, P=0.043], melancholia ( β=0.163, 95% CI=0.073-0.253, P<0.001 ) and fatigue ( β=0.138, 95% CI=0.071-0.205, P<0.001) were still associated with the decline of everyday cognition. Conclusion:The symptoms of hot flashes/sweating, melancholia and fatigue are independent risk factors for the decline of everyday cognition in perimenopausal and postmenopausal women.

Objective:To evaluate the relationship between menopausal symptoms and everyday cognition in perimenopausal and postmenopausal women.Methods:A cross-sectional study was used to analyze the clinical data of 314 women who first visited the menopausal clinic of Department of Gynecology and Obstetrics of Shanghai Jiao Tong University Affiliated Sixth People's Hospital from October 2018 to October 2020. The modified Kupperman menopausal index (KMI) scale was used to assess the prevalence and severity of menopausal symptoms, and a short version of everyday cognition (ECog) scale was used to evaluate everyday cognition. Multiple linear regression was used to analyze the relationship between menopausal symptoms and everyday cognition in perimenopausal and postmenopausal women.Results:The ECog score of 314 patients was 1.50±0.49. The score of KMI was 12.67±7.88, 74.8% (235/314) of the patients had menopausal symptoms, and 34.7% (109/314) had moderate to severe menopausal symptoms. Compared with asymptomatic patients (ECog scores: 1.27±0.29), symptomatic patients had higher ECog scores (mild symptoms group: 1.48±0.47, P=0.005; moderate to severe symptoms group: 1.69±0.55, P<0.001) . Correlation analysis showed that the five symptoms of KMI (hot flashes/sweating, insomnia, irritability, melancholia and fatigue) were associated with the decline of everyday cognition ( P<0.001). Multiple linear regression analysis showed that the symptoms of hot flashes/sweating [ β=0.060, 95% confidence interval ( CI) =0.002-0.117, P=0.043], melancholia ( β=0.163, 95% CI=0.073-0.253, P<0.001 ) and fatigue ( β=0.138, 95% CI=0.071-0.205, P<0.001) were still associated with the decline of everyday cognition. Conclusion:The symptoms of hot flashes/sweating, melancholia and fatigue are independent risk factors for the decline of everyday cognition in perimenopausal and postmenopausal women.

To develop a quality control checklist for the prevention and control of coronavirus disease 2019 （COVID-19） in fever clinic and isolation ward of the general hospital and to assess its application. Based on the relevant prevention and control plans and technical guidelines for COVID-19，Delphi method was used to identity items for evaluation，and a quality control checklist for the prevention and control of COVID-19 in the fever clinic and isolation ward was developed in Sir Run Run Shaw Hospital. The checklists included 8 dimensions and 32 items for fever clinic，7 dimensions and 27 items for the isolation ward. The appointed inspectors conducted daily quality control for each shift with this checklist. The expert authority coefficient was 0.88，the mean of the importance of each index in the quality control table was not less than 4.8，and the coefficient of variation was not more than 0.07. During the entire February 2020，8 problems were found and rectified on-the-spot with the application of the checklist. Quality inspection rate was 100% in both isolation wards and fever clinic. The compliance rate and accuracy rate of hand hygiene were 100%; the correct rate of wearing and removing protective equipment increased from 96% to 100%. During the same period，a total of 1915 patients were admitted to the fever clinic，including 191 suspected patients （all were isolated in the hospital，3 were confirmed）. There were no medical staff infected with COVID-19，no cross infection of patients and their families in the hospital. A quality control checklist for the prevention and control of COVID-19 has been developed and applied in the isolation wards and fever clinic，which plays an important role in preventing nosocomial infection.
COVID-19 ; Checklist ; Fever ; Hospitals, General ; Humans ; SARS-CoV-2