BACKGROUND: Molecular subtyping is an essential complementarity after pathological analyses for targeted therapy. This study aimed to investigate the consistency of next-generation sequencing (NGS) results between circulating tumor DNA (ctDNA)-based and tissue-based in non-small cell lung cancer (NSCLC) and identify the patient characteristics that favor ctDNA testing. METHODS: Patients who diagnosed with NSCLC and received both ctDNA- and cancer tissue-based NGS before surgery or systemic treatment in Lung Cancer Center, Sichuan University West China Hospital between December 2017 and August 2022 were enrolled. A 425-cancer panel with a HiSeq 4000 NGS platform was used for NGS. The unweighted Cohen's kappa coefficient was employed to discriminate the high-concordance group from the low-concordance group with a cutoff value of 0.6. Six machine learning models were used to identify patient characteristics that relate to high concordance between ctDNA-based and tissue-based NGS. RESULTS: A total of 85 patients were enrolled, of which 22.4% (19/85) had stage III disease and 56.5% (48/85) had stage IV disease. Forty-four patients (51.8%) showed consistent gene mutation types between ctDNA-based and tissue-based NGS, while one patient (1.2%) tested negative in both approaches. Patients with advanced diseases and metastases to other organs would be suitable for the ctDNA-based NGS, and the generalized linear model showed that T stage, M stage, and tumor mutation burden were the critical discriminators to predict the consistency of results between ctDNA-based and tissue-based NGS. CONCLUSION ctDNA-based NGS showed comparable detection performance in the targeted gene mutations compared with tissue-based NGS, and it could be considered in advanced or metastatic NSCLC.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Circulating Tumor DNA/blood* ; High-Throughput Nucleotide Sequencing/methods* ; Female ; Male ; Lung Neoplasms/pathology* ; Middle Aged ; Mutation/genetics* ; Aged ; Adult ; Aged, 80 and over

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

The thalamic reticular nucleus (TRN) plays a crucial role in regulating sensory encoding, even at the earliest stages of visual processing, as evidenced by numerous studies. Orientation selectivity, a vital neural response, is essential for detecting objects through edge perception. Here, we demonstrate that somatostatin (SOM)-expressing and parvalbumin (PV)-expressing neurons in the TRN project to the dorsal lateral geniculate nucleus and modulate orientation selectivity and the capacity for visual information processing in the primary visual cortex (V1). These findings show that SOM-positive and PV-positive neurons in the TRN are powerful modulators of visual information encoding in V1, revealing a novel role for this thalamic nucleus in influencing visual processing.
Animals ; Somatostatin/metabolism* ; Parvalbumins/metabolism* ; Neurons/physiology* ; Thalamic Nuclei/physiology* ; Visual Pathways/physiology* ; Mice ; Mice, Inbred C57BL ; Visual Perception/physiology* ; Male ; Mice, Transgenic ; Visual Cortex/physiology* ; Primary Visual Cortex/cytology*

Ferroptosis of chondrocytes is a significant contributor to osteoarthritis (OA), for which there is still a lack of safe and effective therapeutic drugs targeting ferroptosis. Here, we screen for anti-ferroptotic drugs in Food and Drug Administration (FDA)-approved drug library via a high-throughput manner in chondrocytes. We identified a group of FDA-approved anti-ferroptotic drugs, among which vitamin K showed the most powerful protective effect. Further study demonstrated that vitamin K effectively inhibited ferroptosis and alleviated the extracellular matrix (ECM) degradation in chondrocytes. Intra-articular injection of vitamin K inhibited ferroptosis and alleviated OA phenotype in destabilization of the medial meniscus (DMM) mouse model. Mechanistically, transcriptome sequencing and knockdown experiments revealed that the anti-ferroptotic effects of vitamin K depended on growth arrest-specific 6 (Gas6). Furthermore, exogenous expression of Gas6 was found to inhibit ferroptosis through the AXL receptor tyrosine kinase (AXL)/phosphatidylinositol 3-kinase (PI3K)/AKT serine/threonine kinase (AKT) axis. Together, we demonstrate that vitamin K inhibits ferroptosis and alleviates OA progression via enhancing Gas6 expression and its downstream pathway of AXL/PI3K/AKT axis, indicating vitamin K as well as Gas6 to serve as a potential therapeutic target for OA and other ferroptosis-related diseases.

Ferroptosis of chondrocytes is a significant contributor to osteoarthritis(OA),for which there is still a lack of safe and effective therapeutic drugs targeting ferroptosis.Here,we screen for anti-ferroptotic drugs in Food and Drug Administration(FDA)-approved drug library via a high-throughput manner in chondrocytes.We identified a group of FDA-approved anti-ferroptotic drugs,among which vitamin K showed the most powerful protective effect.Further study demonstrated that vitamin K effectively inhibited ferroptosis and alleviated the extracellular matrix(ECM)degradation in chondrocytes.Intra-articular injection of vitamin K inhibited ferroptosis and alleviated OA phenotype in destabilization of the medial meniscus(DMM)mouse model.Mechanistically,transcriptome sequencing and knockdown experiments revealed that the anti-ferroptotic effects of vitamin K depended on growth arrest-specific 6(Gas6).Furthermore,exogenous expression of Gas6 was found to inhibit ferroptosis through the AXL receptor tyrosine kinase(AXL)/phosphatidylinositol 3-kinase(PI3K)/AKT serine/threonine kinase(AKT)axis.Together,we demonstrate that vitamin K inhibits ferroptosis and alleviates OA progression via enhancing Gas6 expression and its downstream pathway of AXL/PI3K/AKT axis,indicating vitamin K as well as Gas6 to serve as a potential therapeutic target for OA and other ferroptosis-related diseases.

Postoperative pulmonary complications and anastomotic leakage are unfavorable prognostic factors in patients with esophageal carcinoma.However,the prognostic importance of pulmonary complications and anastomotic leakage after neoadjuvant treatment in these patients remains unclear.This study aimed to determine the effect of postoperative pulmonary complications and anastomotic leakage on long-term survival after neoadjuvant therapy for esophageal cancer.Our study were recruited 441 consecutive patients who had curative resection following neoadjuvant treatment for esophageal cancer in our institution from 2011-2021.The clinicopathological characteristics and prognosis of these patients were studied in terms of postoperative pulmonary complications and anastomotic leaking.Survival was analyzed using the log-rank test and multivariable Cox regression analysis.Postoperative pulmonary complications and anastomotic leakage were present in 23.8％(n=105)and 5.2％(n=23)of esophageal cancer after neoadjuvant therapy,respectively.In the univariate analyses,pulmonary complications were associated with shorter disease-free survival,while anastomotic leakage was associated with shorter overall survival.Multivariable analysis revealed that pulmonary complications after neoadjuvant therapy were independent adverse prognostic factors for disease-free survival.Taken together,postoperative pulmonary complications and anastomotic leakage ware significantly negatively correlated with disease-free and overall survival,respectively.And the postoperative pulmonary complication is an independent poor prognostic factor of disease-free survival for esophageal cancer following neoadjuvant treatment.

Objective To explore the differences in scapular motion and shoulder function between patients suffering from rotator cuff tears(RCT)with and without type Ⅲ scapular dyskinesis(SD).Meth-ods Between September 2021 and March 2023,sixteen patients suffering from rotator cuff tears with SD(SD group)and 17 counterparts without SD(non-SD group)were recruited from the Sports Hospital of the General Administration of Sport of China.Their scapular motion was assessed by measuring three parameters in the X-rays,including scapular spine line(LSS),scapular upward rotation angle(SU-RA),and coracoid upward shift distance(CUSD).Moreover,their shoulder range of motion in flexion,abduction and external rotation were recorded,and further evaluated using the Pain Visual Analog Scale(VAS)and American Shoulder and Elbow Surgeons Score(ASES).Results No significant differenc-es were found between the two groups in the average score of SURA,CUSD and LSS at 0°～30° shoul-der abduction,or in that of CUSD and LSS at 60°～90°shoulder abduction.However,the average SU-RA score of the SD group at 60°～90°shoulder abduction was significantly greater than the other group(P<0.05).The shoulder ranges of motion during active flexion,abduction and external rotation were significantly smaller in the SD group than in the non-SD group(P<0.05).Moreover,the average VAS score in the SD group was significantly higher than the non-SD group(P<0.05),while the average ASES score was significantly lower than the latter group(P<0.05).Conclusions RCT patients type III SD exhibits greater scapular upward rotation during shoulder abduction compared to those without SD.Moreover,the former patients suffer from more severe pain and have worse shoulder range of motion and functional performance than the latter.

Malignant bowel obstruction(MBO) is a common complication of advanced ventral-pelvic tumors, with poor overall prognosis and seriously affecting the quality of life of these patients. Numerous factors are involved in the pathogenesis of MBO and there is no standard treatment. Most MBO patients were not suitable for surgery. The use of chemotherapy and total parenteral nutrition(TPN) is also controversial. The best treatment is still supportive treatment, including drug treatment and gastrointestinal decompression. Here we reported a case diagnosed as MBO. Transnasal ileus cathete was inserted immediately when hospitalizated after diagnosed 5 days. The symptom was partial relieved after conservative treatment. The abdomen CT scan suggested the sign of closed-loop obstruction. Then, an exploratory laparotomy was performed and found proximal small intestine obviously decompressed with transnasal ileus catheter. However, the tumor caused the closed-loop obstruction of terminal ileum and colon, then the transverse colostomy and terminal ileostomy were performed. After treatment, the patient recovered and was discharged from the hospital. In conclusion, the use of transnasal ileus catheter combined with surgery in MBO, could improve the accuracy of condition judgment, increase the feasibility and safety of surgery, and achieved the goals of treatment expected, and might be a potential treatment regimen for MBO.

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.

Objective To explore the feasibility of CT evaluation of myocardial strain in patients undergoing transcatheter aortic valve replacement(TAVR)with cardiac magnetic resonance(CMR)as the reference standard.Methods A retrospective study was conducted on 43 patients undergoing cardiac CT examination before TAVR in our hospital from June to December 2022.The cardiac CT and CMR data were collected,and myocardial strain parameters were measured.The cardiac function and strain parameters based on cardiac CT were compared with the parame-ters of CMR,including left ventricular ejection fraction(LVEF),left ventricular end-diastolic vol-ume(LVEDV),left ventricular end-systolic volume(LVESV),diastolic myocardial mass(MASS),cardiac output,global peak longitudinal strain(GPLS),global peak circumferential strain(GPCS)and global peak radial strain(GPRS).Results The values of LVEF and cardiac output derived from cardiac CT were close to those from CMR(P＞0.05),while the values of LVEDV,LVESV and MASS from cardiac CT were significantly higher than those from CMR,and those of GPLS,GCPS and GRPS from cardiac CT were obviously lower than those from CMR(P＜0.01).Pearson correlation analysis showed that the LVEF,LVEDV,LVESV,MASS,GCPS and GRPS derived from cardiac CT had strongly positive correlation with those from CMR(r=0.908,r=0.917,r=0.947,r=0.954,r=0.856,r=0.864,P＜0.01),while the cardiac output and GPLS had a moderate positive correlation(r=0.763,r=0.656,P＜0.01).Bland-Altman analysis showed that the LVEF,cardiac output,GPLS,GCPS and GRPS derived from cardiac CT had small biases with those from CMR.Conclusion Cardiac CT quantitative evaluation of cardiac function and strain is feasible in patients undergoing TAVR,and the cardiac function parameters of cardiac CT have good agreement with those from CMR,while the myocardial strain parameters are lower than those from CMR.