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 To investigate the effect of miR-206 on LPS-induced apoptosis and inflamma-tory response in H9c2 cardiomyocytes.Methods An inflammatory injury model was established in H9c2 cells with treatment of 10 μg/ml LPS.Then the cells were divided into control group,LPS group,LPS1 group(LPS+anti-negative control),LPS2 group(LPS+anti-miR-206),LPS3 group(LPS+miR-206 negative control),and LPS4 group(LPS+miR-206 mimic).The LPS1,LPS2,LPS3,and LPS4 groups(n=3)were transfected with anti-negative control,anti-miR-206 se-quence,miR-negative control,or miR-206 mimic sequence,using Lipofectamine 3000 reagent.CCK-8 and EdU assays were used to detect cell proliferation,Caspase-Glo 3/7 activity kit was em-ployed to measure Caspase-3/7 activity,and ELISA was utilized to test the secretion of inflamma-tory cytokines,TNF-α,IL-1β,and IL-6 in the cells.Results Compared with the control group,the LPS group had significantly reduced cell survival rate and EDU-positive cell rate,while elevated miR-206 expression,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65(P＜0.05,P＜0.01).The expression of miR-206,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65 were obviously reduced,while the cell survival rate and EDU-positive cell rate were notably increased in the LPS 2 group than the LPS group(P＜0.05).In the LPS 4 group,the activity of Caspase-3/7 was remarkably stronger than the LPS group(1586±58 vs 816±51,P＜0.05).Conclusion Suppression of miR-206 can effectively inhibit LPS-induced apoptosis and inflammatory response in H9c2 cardiomyocytes.

Objective:To evaluate the efficacy and safety of nebulized inhalation of recombinant human interferon (IFN) α1b injection in the treatment of respiratory syncytial virus (RSV) associated lower respiratory tract infections (pneumonia and bronchiolitis) in children.Methods:A randomized, double-blind, parallel, placebo-controlled add-on design was used.Children with pneumonia or bronchiolitis aged 2 months to 5 years who tested positive for RSV antigen within 72 hours of onset from 30 clinical trial sites including Beijing Children′s Hospital, Capital Medical University between February 2021 and December 2022 were included in this study and randomly divided into 2 groups at a ratio of 1∶1 based on a stratified-block method.Both groups received basic treatments such as cough control, asthma relieving, expectorant treatment, fever reduction, oxygen therapy, etc.The experimental group received additional nebulized inhalation of IFN α1b injection at a dose of 2.0 μg/(kg·time), twice a day.The control group received nebulized inhalation of placebo twice a day.Clinical efficacy was evaluated based on indicators such as the duration of clinical symptoms and signs, and the Kaplan-Meier method was used to calculate the median and 95% CI of the duration of clinical symptoms and signs.The Log-rank test was used to compared data between groups.Safety was assessed through the incidence of adverse reactions and laboratory tests, and the Chi-square test was used to analyze the difference between groups. Results:There were 123 children in the experimental group and 122 children in the control group.The median durations of all the 5 clinical symptoms and signs [including shortness of breath, wheezing, dyspnea (visible retractions), decreased transcutaneous oxygen saturation, and abnormal mental state] in the experimental group after treatment were slightly shortened than those in the control group [2.7 d(95% CI: 1.9-3.0 d)] vs.[2.9 d(95% CI: 2.6-3.6 d), P=0.027].The improvement in dyspnea (retractions) was especially pronounced in the experimental group, with a relief rate of 50.0% (0, 100%) on the first day of administration[compared with 0 (0, 50.0%) in the control group ( Z=2.002, P=0.025)].The median duration of dyspnea in the experimental group was nearly 1 day shorter than that in the control group [1.0 d(95% CI: 0.7-1.7 d) vs.1.8 d(95% CI: 1.0-2.5 d), P=0.046].There were no significant difference in hospital stay [6.0(5.0, 8.0) d vs.6.5(5.0, 8.0) d, Z=0.675, P=0.500], oxygen therapy duration [32.0(14.0, 96.3) h vs.39.0 (24.0, 83.2) h, Z=0.094, P=0.925], the recovery rate from clinical symptoms during treatment [(105/106, 99.1%) vs.(96/101, 95.0%)], and recurrence rate [(0/106, 0) vs.(2/101, 2.0%)] between the 2 groups (all P>0.05).However, the above-mentioned four indicators in the experimental group showed a trend of clinical benefits.The quantitative virus detection results showed that the RSV viral load in both groups decreased after treatment compared to before treatment.After 2 days of treatment, the decline rate of RSV viral load from the baseline was 0.90 lg copies/(mL·d) in the experimental group and 0.25 lg copies/(mL·d)in the control group, with a statistically significant difference ( P<0.05).Furthermore, there was no statistically significant difference in the incidence of adverse reactions between the 2 groups ( P>0.05).Importantly, no drug-related serious adverse reactions occurred in both groups. Conclusions:The nebulized inhalation therapy of IFN α1b demonstrates efficacy and safety in treating pediatric RSV associated lower respiratory tract infections.It particularly offers outstanding clinical therapeutic value for severe children.

Objective:To evaluate the efficacy and safety of nebulized inhalation of recombinant human interferon (IFN) α1b injection in the treatment of respiratory syncytial virus (RSV) associated lower respiratory tract infections (pneumonia and bronchiolitis) in children.Methods:A randomized, double-blind, parallel, placebo-controlled add-on design was used.Children with pneumonia or bronchiolitis aged 2 months to 5 years who tested positive for RSV antigen within 72 hours of onset from 30 clinical trial sites including Beijing Children′s Hospital, Capital Medical University between February 2021 and December 2022 were included in this study and randomly divided into 2 groups at a ratio of 1∶1 based on a stratified-block method.Both groups received basic treatments such as cough control, asthma relieving, expectorant treatment, fever reduction, oxygen therapy, etc.The experimental group received additional nebulized inhalation of IFN α1b injection at a dose of 2.0 μg/(kg·time), twice a day.The control group received nebulized inhalation of placebo twice a day.Clinical efficacy was evaluated based on indicators such as the duration of clinical symptoms and signs, and the Kaplan-Meier method was used to calculate the median and 95% CI of the duration of clinical symptoms and signs.The Log-rank test was used to compared data between groups.Safety was assessed through the incidence of adverse reactions and laboratory tests, and the Chi-square test was used to analyze the difference between groups. Results:There were 123 children in the experimental group and 122 children in the control group.The median durations of all the 5 clinical symptoms and signs [including shortness of breath, wheezing, dyspnea (visible retractions), decreased transcutaneous oxygen saturation, and abnormal mental state] in the experimental group after treatment were slightly shortened than those in the control group [2.7 d(95% CI: 1.9-3.0 d)] vs.[2.9 d(95% CI: 2.6-3.6 d), P=0.027].The improvement in dyspnea (retractions) was especially pronounced in the experimental group, with a relief rate of 50.0% (0, 100%) on the first day of administration[compared with 0 (0, 50.0%) in the control group ( Z=2.002, P=0.025)].The median duration of dyspnea in the experimental group was nearly 1 day shorter than that in the control group [1.0 d(95% CI: 0.7-1.7 d) vs.1.8 d(95% CI: 1.0-2.5 d), P=0.046].There were no significant difference in hospital stay [6.0(5.0, 8.0) d vs.6.5(5.0, 8.0) d, Z=0.675, P=0.500], oxygen therapy duration [32.0(14.0, 96.3) h vs.39.0 (24.0, 83.2) h, Z=0.094, P=0.925], the recovery rate from clinical symptoms during treatment [(105/106, 99.1%) vs.(96/101, 95.0%)], and recurrence rate [(0/106, 0) vs.(2/101, 2.0%)] between the 2 groups (all P>0.05).However, the above-mentioned four indicators in the experimental group showed a trend of clinical benefits.The quantitative virus detection results showed that the RSV viral load in both groups decreased after treatment compared to before treatment.After 2 days of treatment, the decline rate of RSV viral load from the baseline was 0.90 lg copies/(mL·d) in the experimental group and 0.25 lg copies/(mL·d)in the control group, with a statistically significant difference ( P<0.05).Furthermore, there was no statistically significant difference in the incidence of adverse reactions between the 2 groups ( P>0.05).Importantly, no drug-related serious adverse reactions occurred in both groups. Conclusions:The nebulized inhalation therapy of IFN α1b demonstrates efficacy and safety in treating pediatric RSV associated lower respiratory tract infections.It particularly offers outstanding clinical therapeutic value for severe children.

Objective To investigate the effect of miR-206 on LPS-induced apoptosis and inflamma-tory response in H9c2 cardiomyocytes.Methods An inflammatory injury model was established in H9c2 cells with treatment of 10 μg/ml LPS.Then the cells were divided into control group,LPS group,LPS1 group(LPS+anti-negative control),LPS2 group(LPS+anti-miR-206),LPS3 group(LPS+miR-206 negative control),and LPS4 group(LPS+miR-206 mimic).The LPS1,LPS2,LPS3,and LPS4 groups(n=3)were transfected with anti-negative control,anti-miR-206 se-quence,miR-negative control,or miR-206 mimic sequence,using Lipofectamine 3000 reagent.CCK-8 and EdU assays were used to detect cell proliferation,Caspase-Glo 3/7 activity kit was em-ployed to measure Caspase-3/7 activity,and ELISA was utilized to test the secretion of inflamma-tory cytokines,TNF-α,IL-1β,and IL-6 in the cells.Results Compared with the control group,the LPS group had significantly reduced cell survival rate and EDU-positive cell rate,while elevated miR-206 expression,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65(P＜0.05,P＜0.01).The expression of miR-206,Bax/Bcl-2,activated Caspase-3,Caspase-3/7,TNF-α,IL-1β,IL-6,p-p38 and p-p65 were obviously reduced,while the cell survival rate and EDU-positive cell rate were notably increased in the LPS 2 group than the LPS group(P＜0.05).In the LPS 4 group,the activity of Caspase-3/7 was remarkably stronger than the LPS group(1586±58 vs 816±51,P＜0.05).Conclusion Suppression of miR-206 can effectively inhibit LPS-induced apoptosis and inflammatory response in H9c2 cardiomyocytes.

Objective:To explore the effect of enriched environment on the A1/A2 phenotype conversion of astrocytes and cognitive function in rats after ischemia-reperfusion (I/R) in rats. Method:Forty two adult male SD rats (weight 220±20g) were selected for middle cerebral artery occlusion (MCAO) surgery,followed by reperfusion 2 hours later. Three days after operation,30 rats were randomly di-vided into standard environment group (n=15) and enriched environment group (n=15),and 10 rats were se-lected as sham operation group. The enriched environment group was raised in the enriched environment cag-es,and the other two groups were raised in the standard environment. After 21 days,the Bederson score and mNss score were used to detect the behavioral changes of rats in each group,and the Morris water maze was used to detect the cognitive function of rats. Subsequently,western blot analysis was used to analyze the acti-vation of glial fibrillary acidic protein (GFAP),a marker of astrocytes. Real-time PCR and ELISA detect the expression of A1 type astrocyte marker (C3) and A2 type astrocyte marker (S100A10). Hematoxylin eosin (HE) staining was used to observe the pathological changes of cortex around the infarction. TUNEL staining was used to detect apoptosis.Result:Compared with SE group,the expression of GFAP protein in EE group decreased significantly(P<0.05). The expression level of the A1 type astrocyte marker C3 in EE group was significantly lower than that in SE group (P<0.05),while the expression of A2 type astrocyte marker S100A10 was significantly higher than that in SE group (P<0.05). Correspondingly to this result,in the EE group,the secretion of the cyto-kine TNF-α by A1 astrocytes was significantly lower than in the SE group(P<0.05),and the secretion of the cytokine BDNF by A2 astrocytes was significantly higher than in the SE group (P<0.05). TUNEL and HE staining showed that the apoptosis and damage of cells in EE group were less (P<0.05). In addition,the neu-rological ischemia in the EE group was less severe,including significant differences in the Bederson score and mNss score (P<0.01). Compared with the SE group,the rats in the EE group had better cognitive func-tion,characterized by shorter latency (P<0.05),longer residence time in the target quadrant (P<0.01),and more times of crossing the platform (P<0.05) in the water maze experiment.Conclusion:The enriched environment can inhibit the activation of astrocytes,promote the conversion of acti-vated astrocytes to neuroprotective type A2 and inhibit their conversion into neurotoxic type A1,resulting in improving cognitive function after ischemic stroke in rats.

Objective:To explore the effect of enriched environment on the A1/A2 phenotype conversion of astrocytes and cognitive function in rats after ischemia-reperfusion (I/R) in rats. Method:Forty two adult male SD rats (weight 220±20g) were selected for middle cerebral artery occlusion (MCAO) surgery,followed by reperfusion 2 hours later. Three days after operation,30 rats were randomly di-vided into standard environment group (n=15) and enriched environment group (n=15),and 10 rats were se-lected as sham operation group. The enriched environment group was raised in the enriched environment cag-es,and the other two groups were raised in the standard environment. After 21 days,the Bederson score and mNss score were used to detect the behavioral changes of rats in each group,and the Morris water maze was used to detect the cognitive function of rats. Subsequently,western blot analysis was used to analyze the acti-vation of glial fibrillary acidic protein (GFAP),a marker of astrocytes. Real-time PCR and ELISA detect the expression of A1 type astrocyte marker (C3) and A2 type astrocyte marker (S100A10). Hematoxylin eosin (HE) staining was used to observe the pathological changes of cortex around the infarction. TUNEL staining was used to detect apoptosis.Result:Compared with SE group,the expression of GFAP protein in EE group decreased significantly(P<0.05). The expression level of the A1 type astrocyte marker C3 in EE group was significantly lower than that in SE group (P<0.05),while the expression of A2 type astrocyte marker S100A10 was significantly higher than that in SE group (P<0.05). Correspondingly to this result,in the EE group,the secretion of the cyto-kine TNF-α by A1 astrocytes was significantly lower than in the SE group(P<0.05),and the secretion of the cytokine BDNF by A2 astrocytes was significantly higher than in the SE group (P<0.05). TUNEL and HE staining showed that the apoptosis and damage of cells in EE group were less (P<0.05). In addition,the neu-rological ischemia in the EE group was less severe,including significant differences in the Bederson score and mNss score (P<0.01). Compared with the SE group,the rats in the EE group had better cognitive func-tion,characterized by shorter latency (P<0.05),longer residence time in the target quadrant (P<0.01),and more times of crossing the platform (P<0.05) in the water maze experiment.Conclusion:The enriched environment can inhibit the activation of astrocytes,promote the conversion of acti-vated astrocytes to neuroprotective type A2 and inhibit their conversion into neurotoxic type A1,resulting in improving cognitive function after ischemic stroke in rats.

Objective To explore the effect of micro-lectures withadvanced simulation man in improving the practical skills teaching of nursing students, so as to promote the students' post competency. Methods Totally the 186 nursing internswere divided into control group and observation group by random number method with 93 people in each group. The control group used the traditional teaching modein the teaching of practical skills.The observation group used the micro-lectures with high simulation teaching.Comparing the two groups of nursing students comprehensive assessment test simulation results in the theory, skills, scenarios, and nursing students the evaluation of the curriculum. Results The scores of the two groups were all above the qualification line, but the scores of the observation group were significantly higher than those of the control group (P<0.05). The theoretical examinations and the situation simulation comprehensive testswas(77.89 ± 7.79), (75.60 ± 7.92)points in control group, and (93.87 ± 3.90),(92.87 ± 4.08)points in observation group, there was significant difference between two groups (t=17.67, 18.70,all P=0.000). The curriculum evaluation results of improving learning initiative, active curriculum atmosphere, clear operation demonstration, exercise clinical thinking, improve the clinical interest were 81.72％(76/93), 72.04％(67/93), 93.55％(87/93), 60.22％(56/93), 67.74％(63/93)in control group, and 96.77％(90/93), 95.70％(89/93), 100.00％(93/93), 92.47％(86/93), 98.92％(92/93)in observation group, there was significant difference between two groups(χ2=20.39, P=0.016).Conclusions The effect is significant of micro-lectures combined with high simulation of human using in clinical skills teaching. Thismold can conducive to the cultivation of clinical thinking of nursing students, and improve clinical comprehensive ability, and promote the promotion of nursing students post competency.

Objective Based on the principles of laser radiation protection, medical metroiogy and photoelectron technology, an automatic verification device and testing technology to provide verification and performance evaluation for laser protective spectacles and equipments which have the various functions in laser protection have been developed and established. Methods The current system comprises laser source, laser measuring instruments, software of computer detection and control and modules of optics and mechanics used in auxiliary equipment. By use of the verification device and the special test-recording method to be designed and studied by us, the measured data can be automatically acquired, processed, recorded, saved and printed and, furthermore, many parameters on protective performance of the measured laser protective spectacles can be tested and given. Results Laser wavelength of the verification apparatus are 1 064 nm and 532 nm, response range of wavelength is from 0.4 m to 1.1 μm, measuring range of laser energy is from 10-8 J to 0.3 J, measuring range of optical density for laser protective spectacles is from 0.1 to 8.0, stability is 0.21%, measuring uncertainty is 5%(k=2). Conclusion The automatic verification device is steady and reliable. The achieved performance indexes accord with the requirement of national standard on laser protection.