Objective This study aims to achieve high-yield functional expression of the human voltage-gated potassium channel K_V3.1 using an Escherichia coli cell-free protein synthesis system, thereby providing a novel synthetic approach for drug screening, structural analysis and functional characterization of K_V3.1. Methods K_V3.1 was expressed in an Escherichia coli cell-free protein synthesis system for 10 hours in the presence of peptide surfactant A₆K. The secondary structure of K_V3.1 was analyzed by circular dichroism spectroscopy. The potassium channel activity of the recombinant protein liposome K_V3.1-A₆K was investigated using fluorescent dyes Oxonol VI as indicators, which are capable of reflecting alterations in membrane potential. Results Soluble K_V3.1 protein was successfully synthesized, achieving a purified yield of up to 1.2 mg/mL via an Escherichia coli cell-free protein synthesis system. Circular dichroism spectroscopy revealed that K_V3.1 exhibited characteristic α-helical secondary structures. Membrane potential fluorescence assays demonstrated that the K_V3.1-A₆K proteoliposomes, which were reconstructed with surfactant peptide A₆K, exhibited remarkable potassium ion permeability. Conclusion This study successfully achieved high-yield expression of human K_V3.1 with activity using an Escherichia coli-based cell-free protein synthesis system. This innovative method not only significantly enhances the expression yield of K_V3.1, but also maintains its functional activity, thereby establishing a novel and efficient synthetic platform for drug screening and advancing our understanding of structure-function relationships in K_V3.1 research.
Humans ; Escherichia coli/metabolism* ; Shaw Potassium Channels/biosynthesis* ; Cell-Free System ; Circular Dichroism ; Protein Biosynthesis ; Recombinant Proteins/metabolism* ; Membrane Potentials ; Shab Potassium Channels

Objective:To investigate the effects of cervical cold knife conization (CKC) on preterm delivery, other pregnancy complications and neonatal outcomes, and explore the relationship between preterm delivery risk and the depth and volume of conization.Methods:The clinical data and pregnancy outcomes of 272 women who underwent CKC in Peking Union Medical College Hospital from January 2002 to March 2018 (conization group) and 1 647 pregnant women who gave birth in Peking Union Medical College Hospital during January to December 2019 (control group) were collected. The preterm delivery, premature rupture of membranes, other pregnancy complications and neonatal outcomes of the two groups were compared, and the relationship between the depth and volume of conization and the risk of preterm delivery in postoperative singleton pregnancy was analyzed.Results:(1) There were no significant differences between the two groups in delivery age, parity, proportion of singleton pregnancy, proportion of assisted reproductive technology (all P>0.05). (2) The rate of preterm delivery in the conization group was significantly higher than that in the control group [14.8% (39/264) vs 5.7% (91/1 589); χ2=28.397, P<0.001]. There were still significant differences in preterm delivery rates between the two groups at <34 weeks and 34-37 weeks (all P<0.01). There was no significant difference in the incidence of premature rupture of membrane between the two groups [23.5% (62/264) vs 23.4% (372/1 589); χ2=0.001, P=0.979], but the incidence of preterm premature rupture of membrane in the conization group was significantly higher than that in the control group [11.4% (30/264) vs 2.2% (35/1 589); χ2=56.132, P<0.001]. (3) The rate of cesarean section in the conization group was higher than that in the control group [59.6% (162/272) vs 38.8% (639/1 647); χ2=41.377, P<0.001]. The birth weight of preterm infants in the conization group was significantly higher than that in the control group [(2 409±680) vs (2 150±684) g; t=2.184, P=0.030]. However, there were no statistically significant differences in the incidence of gestational diabetes mellitus, hypertensive disorders in pregnancy, the birth weight of full-term infants, incidence of small for gestational age infant and neonatal intensive care unit admission rate between the two groups (all P>0.05). (4) The preterm delivery rates of coning depth >15 mm, cone size ≥2 cm 3 and cone size <2 cm 3 were higher than that in the control group (all P<0.05). When the coning depth ≤15 mm, the preterm delivery rate in the conization group was higher than that in the control group, but there was no significant difference ( P=0.620). The rate of preterm delivery of pregnant women with coning depth >15 mm was significantly higher than those with coning depth ≤15 mm ( RR=3.084, 95% CI: 1.474-6.453; P=0.001). There was no significant difference in the preterm delivery rate between pregnant women with cone size >2 cm 3 and those with cone size ≥2 cm 3 ( RR=1.700, 95% CI: 0.935-3.092; P=0.077). Conclusion:The risk of preterm delivery and preterm premature rupture of membranes in subsequent pregnancies are increased after cervical CKC, and the risk of preterm delivery is positively correlated with the depth of cervical coning.

Lung cancer is the malignant tumor with the highest incidence and mortality among the Chinese.Tumor node metastasis(TNM)staging established by the American Joint Committee on Cancer(AJCC)and International Union Against Cancer(UICC)is a commonly used criterion,but it still has limitations in judging the prognosis of non-small cell lung cancer(NSCLC)patients.With the advantages of real-time and convenient sampling,the immune score based on peripheral blood biomarkers have the ability to predict prognosis and efficacy of NSCLC patients,which have been developed and validated in clinical studies.However,clinical impleruentation of peripheral immune scores is still not widely in NSCLC patients.Therefore,this study introduces and evaluates the 6 peripheral immune scores and reviews the reseach progress of them in the treatment of NSCLC.

Objective To analyze the needs and current situation of home-based exercise rehabilitation for people with disabilities,develop service plans for different types of disabilities,and assess its health benefits for people with severe dis-abilities. Methods Based on the World Health Organization(WHO)Global Action Plan on Physical Activity 2018-2030:More Ac-tive People for a Healthier World and WHO Guidelines on Physical Activity and Sedentary Behavior,functional and individualized home-based exercise rehabilitation plans for people with severe disabilities were developed.Begining from May,2023,a six-month intervention was conducted for 37 people with severe disabilities.They were assessed with International Physical Activity Questionnaire(IPAQ),WHO Disability Assessment Schedule 2.0(WHODAS 2.0),WHO Quality of Life(WHOQOL-BREF)before and after intervention. Results The time of low-intensity leisure physical activity of IPAQ increased after intervention,while the scores of do-mains of cognition,mobility,self-care,getting along,life activities and participation,and total score of WHO-DAS 2.0 decreased,and the scores of physical health,psychological health,independence,and environment di-mensions,and total score of WHOQOL-BREF increased. Conclusion The functioning-oriented and individualized home exercise rehabilitation program can improve the level of physical activity,enhance the level of activity and boost the quality of life for people with severe disabilities.

Objective:To evaluate the effectiveness of hands-on training and video demonstration in training of forceps delivery for residents.Methods:Forty nine residents who were rotating in the obstetrics department of Peking Union Medical College Hospital from 2019 to 2021 were enrolled. The residents were randomly divided into two groups: the instructor group ( n=24) was taught by hands-on training of forceps delivery and the video group ( n=25) was instructed by watching video demonstration. All the trainees completed the self-confidence questionnaire survey, and were evaluated by written tests and objective structured assessment of technical skills scoring system. Results:The scores of self-confidence in each item after the simulation training were higher than those before training in both groups; and there were no significant differences between two groups in the increment of scores(mastering knowledge: 1.54±0.98 vs. 1.40±0.71, U=266.68, P=0.480;mastering operation skills: 1.42±0.93 vs.1.80±0.87, U=233.47, P=0.161; mastering forceps structure: 1.63±1.10 vs. 1.88±0.93, U=261.63, P=0.416; confidence in independent operation: 1.13±0.90 vs. 1.00±1.08, U=287.74, P=0.799; evaluation of simulation training: 0.21±0.51 vs. 0.16±0.55, U=288.27, P=0.776). In the written tests, the scores of the instructor group were significantly higher than those of the video group (83.00±7.18 vs.70.56±10.37; t=4.86, P<0.001). In the practical operation, the instructor group significantly outperformed the video group in items of “right blade placement” (0.71±0.46 vs. 0.20±0.41, U=147.54, P<0.001), “objective total score” (6.17±1.46 vs. 4.72±1.65, U=155.49, P=0.003) and “correct traction” (0.85±0.31 vs. 0.56±0.51, U=213.86, P=0.036). Conclusion:Training delivered via hands-on instruction and demonstration was generally more effective than that delivered via video, although both groups show a increased self-confidence in learning and performing forceps delivery.

Pulmonary blast injury has become the main type of trauma in modern warfare, characterized by externally mild injuries but internally severe injuries, rapid disease progression, and a high rate of early death. The injury is complicated in clinical practice, often with multiple and compound injuries. Currently, there is a lack of effective protective materials, accurate injury detection instrument and portable monitoring and transportation equipment, standardized clinical treatment guidelines in various medical centers, and evidence-based guidelines at home and abroad, resulting in a high mortality in clinlcal practice. Therefore, the Trauma Branch of Chinese Medical Association and the Editorial Committee of Chinese Journal of Trauma organized military and civilian experts in related fields such as thoracic surgery and traumatic surgery to jointly develop the Clinical treatment guideline for pulmonary blast injury ( version 2023) by combining evidence for effectiveness and clinical first-line treatment experience. This guideline provided 16 recommended opinions surrounding definition, characteristics, pre-hospital diagnosis and treatment, and in-hospital treatment of pulmonary blast injury, hoping to provide a basis for the clinical treatment in hospitals at different levels.

ObjectiveTo observe the effect of ginsenoside Rg₁ （G-Rg₁） on the biological activity of cryopreserved Schwann cells （SCs） of the rat sciatic nerve and explore the feasibility of G-Rg₁ in reducing the cryopreservation-induced injury in SCs. MethodBilateral sciatic nerves of SD rats were randomly divided into a fresh group， a blank group， and five G-Rg₁ groups of different doses （1×10^-7， 1×10^-6， 1×10^-5， 1×10^-4， and 1×10^-3 mol·L^-1）. The nerves in the blank group and the G-Rg₁ groups were preserved in liquid nitrogen solutions containing 0， 1×10^-7， 1×10^-6， 1×10^-5， 1×10^-4， and 1×10^-3 mol·L^-1 G-Rg₁ for four weeks. The apoptosis of SCs was detected by TdT-mediated dUTP-biotin nick end labeling （TUNEL）/S100 immunofluorescence staining. The expression of cysteinyl aspartate-specific protease （Caspase）-9， Caspase-3， major histocompatibility complex （MHC）-Ⅰ， and MHC-Ⅱ was detected by Western blot. Subsequently， all nerves were cultured in the incubator at 37 ℃ with 5% CO₂ for 7 days. The expression of glial cell line-derived neurotrophic factor （GDNF） and nerve growth factor （NGF） was detected by Western blot. In addition， the above cryopreserved nerves in the blank group and the 1×10^-6， 1×10^-5， and 1×10^-4 mol·L^-1 G-Rg₁ groups were transplanted to the Wistar rats by allografting （blank transplantation group and the 1×10^-6， 1×10^-5， and 1×10^-4 mol·L^-1 G-Rg₁ transplantation groups）， and fresh sciatic nerve allograft and isograft control group were set up. Sixteen weeks after transplantation， compound muscle action potential （CMAP） and nerve conduction velocity （NCV） were measured by electrophysiology. Nerve filament （NF）200 immunofluorescence staining， transmission electron microscopy， and toluidine blue staining were used to analyze the histology of the regenerated nerves. ResultCompared with the fresh group， the blank group and the G-Rg₁ groups showed increased expression of Caspase-9， Caspase-3， and the apoptosis of SCs （P<0.05，P<0.01） and decreased expression of GDNF， NGF， MHC-Ⅰ， and MHC-Ⅱ （P<0.01）. Compared with the results in the blank group， the expression of Caspase-9 and Caspase-3 decreased in the 1×10^-7， 1×10^-6， 1×10^-5，1×10^-4 mol·L^-1 G-Rg₁ groups （P<0.01）， and the apoptosis of SCs was reduced in the 1×10^-7-1×10^-4 mol·L^-1 G-Rg₁ groups（P<0.05，P<0.01） and increased in the 1×10^-3 mol·L^-1 group （P<0.05）， while the expression of GDNF and NGF increased in the 1×10^-7， 1×10^-6， 1×10^-5，1×10^-4 mol·L^-1 G-Rg₁ groups and decreased in the 1×10^-3 mol·L^-1 group （P<0.05）. There was no statistical significance in the expression of MHC-Ⅰ and MHC-Ⅱ between the blank group and the G-Rg₁ groups. Compared with the 1×10^-7 mol·L^-1 and 1×10^-3 mol·L^-1 G-Rg₁ groups， the 1×10^-6 1×10^-5， 1×10^-4 mol·L^-1 G-Rg₁ groups showed decreased expression of Caspase-3 and the apoptosis of SCs （P<0.05，P<0.01） and increased expression of GDNF and NGF （P<0.05，P<0.01）. There was no statistical significance in MHC-Ⅰ and MHC-Ⅱ expression among G-Rg₁ groups. Sixteen weeks after transplantation， compared with the isograft group， the blank transplantation group and the G-Rg₁ transplantation groups showed decreased CMAP， NCV， myelin sheath thickness， and number of myelinated nerve fibers （P<0.01）， and the 1×10^-6 and 1×10^-4 mol·L^-1 G-Rg₁ transplantation groups showed decreased NF200 （P<0.01）. Compared with the allograft group， the blank transplantation group and the G-Rg₁ transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Compared with the blank transplantation group， the G-Rg₁ transplantation groups showed increased CMAP， NCV， NF200， myelin sheath thickness， and number of myelinated nerve fibers （P<0.05，P<0.01）. Among all groups of G-Rg₁ transplantation， each index of the 1×10^-5 mol·L^-1 G-Rg₁ transplantation group was superior to that of the 1×10^-4 and 1×10^-6 mol·L^-1 G-Rg₁ transplantation group （P<0.05）. ConclusionG-Rg₁ at a certain centration can maintain the biological activity of cryopreserved SCs of rat sciatic nerve， alleviate the cryopreservation-induced injury of rat sciatic nerve， and promote nerve regeneration after allograft.

Objective    To evaluate the impact of different surgical strategies for moderate functional mitral regurgitation (FMR) at the time of aortic valve replacement (AVR) on patients' prognosis. Methods    A total of 118 AVR patients, including 84 males and 34 females, aged 58.1±12.4 years, who were complicated with moderate FMR were retrospectively recruited. Patients were divided into three groups according to the treatment strategy of mitral valve: a group A (no intervention, n=11), a group B (mitral valve repair, n=51) and a group C (mitral valve replacement, n=56). The primary endpoint was the early and mid-term survival of the patients, and the secondary endpoint was the improvement of FMR. Results    The median follow-up time was 29.5 months. Five patients died perioperatively, all of whom were from the group C. Early postoperative FMR improvement rates in the group A and group B were 90.9% and 94.1% (P=0.694). The mid-term mortality in the three groups were 0.0%, 5.9% and 3.9%, respectively (P=0.264), while the incidences of major cardiovascular and cerebrovascular events were 0.0%, 9.8% and 17.7%, respectively (P=0.230). Improvements of FMR in the group A and group B were 100.0% and 94.3% at the mid-term follow-up (P>0.05). Conclusion    For patients receiving AVR with moderate FMR, conservative treatment or concurrent repair of mitral valve may be more reasonable, while mitral valve replacement may increase the incidence of early and mid-term adverse events.

Objective:To explore the risk factors influencing the prognosis for patients with hepatocellular carcinoma (HCC) recurrence after liver transplantation and summarize the relevant diagnostic and therapeutic experiences.Methods:The clinicopathological features with diagnosis and treatment plan of 102 recurrent HCC patients fulfilling the Fudan Criteria were compared for survival rate (univariate analysis) and independent prognostic indicators were obtained by Cox multivariate analysis.Results:The 1/3/5-year overall survival rates were 92.2%, 48.6% and 34.6% and the 1/3/5-year survival rates with tumor were 63.2%, 31.0% and 16.7% respectively. Cox regression analysis indicated that patient age, whether tumor can be surgically resected or not and personalized diagnostic & therapeutic plan based upon targeted therapy were independent prognostic factors affecting the overall survival rates and survival rates with tumor.Conclusions:Although HCC recurrence and metastasis after liver transplantation seriously influence patient prognosis, satisfactory outcomes may be obtained for some patients through active, effective and precise managements.

Objective:To investigate the cognitive status of the elderly in rural areas and explore the characteristics and influencing factors of subjective cognitive decline (SCD).Methods:A baseline survey was conducted among 5 765 rural elderly people aged 60 years old or above from March to September 2018.Subjective cognitive decline questionnaire(SCD-Q9), mini-mental state examination(MMSE), verbal fluency test (VFT), Chinese auditory verbal learning test (CAVLT), digital span test(DST)and activities of daily living(ADL)were used in the survey.The result of the survey indicated that there were 2 654 subjects with SCD (SCD group) and 1 008 subjects with normal cognitive function (NC group). Social support rating scale (SSRS) and short version of geriatric depression scale-15(GDS-15) were used to evaluate their psychosocial status.Descriptive analysis and Logistic regression analysis were conducted by SPSS 26.0 software.Results:(1) Compared with NC group, the SCD group had the following characteristics: delayed recognition rate(8.25 ±2.51), (12.38 ±2.53), reverse digit span (2.63±1.37), (3.69±1.45), social support (69.81±8.71), (64.40±9.44), GDS-15 (2.27±2.63), (1.31±2.17), and the differences were statistically significant (all P<0.05). However, there were no significant differences in the following characteristics: MMSE score (21.62±5.73), (21.47±5.84), speech fluency (27.80±7.35), (28.25±7.56), ADL score (20.70±1.35), (20.77±1.30), all P>0.05.(2) There were no significant differences in diet structure, blood glucose, blood lipid, cerebrovascular disease, diabetes, epilepsy and coronary heart disease between SCD group and NC group (all P>0.05). (3)SCD was mainly affected by age( β=0.06, OR=2.29, 95% CI=1.09-4.85), depression( β=-0.01, OR=2.96, 95% CI=0.68-4.94), hypertension( β=-0.17, OR=1.89, 95% CI=1.11-2.15), and low level of social support( β=2.07, OR=1.49, 95% CI=1.32-2.12) (all P<0.05). Conclusion:The scores of delayed recognition and reverse digit span in patients with SCD are lower than those with normal cognitive function.The other objective cognitive functions are basically normal.Old age, low social support level, depression, low education level and hypertension are the risk factors of SCD.