Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

Objective:To evaluate the diagnostic value of coronary angiography-derived fractional flow reserve (FFR) and index of microcirculatory resistance (IMR) for identifying coronary functional abnormalities.Methods:This diagnostic study enrolled patients with clinically suspected or diagnosed coronary artery disease who underwent coronary angiography at Beijing Anzhen Hospital, TEDA International Cardiovascular Hospital, and Qilu Hospital of Shandong University between December 2021 and June 2022. All enrolled patients successfully underwent invasive wire-based FFR and IMR measurements during angiography. In a core laboratory, FFR and IMR for the target vessels were measured using artificial intelligence technology based on coronary angiographic images. Spearman correlation analysis was used to evaluate the correlation between angiography-derived FFR and wire-based FFR, and between angiography-derived IMR and wire-based IMR. Coronary hemodynamic abnormality was defined as FFR≤0.80; the diagnostic performance of angiography-derived FFR for identifying this abnormality was evaluated. Microcirculatory dysfunction was defined as IMR≥25; the diagnostic performance of angiography-derived IMR for identifying microcirculatory dysfunction was evaluated.Results:A total of 181 patients, aged (60.6±8.8) years, with 62 (34.3%) females, and 181 target vessels were included in the final analysis. Angiography-derived FFR showed a significant positive correlation with wire-based FFR ( r=0.78, P<0.001). For identifying coronary hemodynamic abnormality, angiography-derived FFR showed an accuracy of 89.0%, sensitivity of 88.8%, specificity of 89.1%, positive predictive value (PPV) of 88.8%, negative predictive value (NPV) of 89.1%, and an area under the receiver operating characteristic curve ( AUC) of 0.88. Angiography-derived IMR showed a significant positive correlation with wire-based IMR ( r=0.93, P<0.001). For identifying microcirculatory dysfunction, angiography-derived IMR demonstrated an accuracy of 89.5%, sensitivity of 86.8%, specificity of 90.2%, PPV of 70.2%, NPV of 96.3%, and an AUC of 0.95. Conclusion:Angiography-derived FFR and IMR exhibit strong correlations with their invasive wire-based counterparts and demonstrate high diagnostic value for assessing coronary hemodynamics and coronary microcirculatory function.

Objective To investigate the efficacy and mechanism of the combination of cryoablation and Yangfei establish the Lewis lung cancer mouse model,and randomly divided into model group,cryoablation group,cryoablation+cisplatin group,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups,with 5 mice in each group.Sham-operation was performed in the model group,cryoablation+cisplatin group were given intraperitoneal injection of cisplatin 3 mg/kg after cryoablation,cryoablation+Yangfei Prescription low-,medium-and high-dosage groups were given 1.64,3.28,6.56 g/kg Yangfei Prescription by gavage after cryoablation respectively,and the model group was given equal volume of normal saline by gavage for 14 consecutive days.Tumor volume and tumor mass were measured,the morphology of tumor tissue was observed by HE staining,the expression of Ki-67 protein in tumor tissue was detected by immunohistochemistry.The differentially expressed genes in tumor tissues after cryoablation combined with Yangfei Prescription intervention were analyzed by whole transcriptome sequencing,GO and KEGG pathway enrichment analysis was performed on differentially expressed genes,and the lncRNA/circNA-mediated ceRNA network was constructed.Differentially expressed genes were verified using RT-qPCR.Results The main active components of Yangfei Prescription were terpenoids,flavonoids and aldehydes.Compared with the model group,tumor volume and tumor mass decreased(P<0.05)in cryoablation+Yangfei Prescription low-,medium-and high-dosage groups and cryoablation+cisplatin group;cryoablation combined with Yangfei Prescription could destroy the structure of tumor tissue and inhibit cell proliferation(P<0.05).A total of 1 585 differentially expressed genes(1 160 mRNA,225 lncRNA,155 miRNA and 45 circRNA)of the model group and cryoablation+Yangfei Prescription high-doseage group were obtained.GO enrichment analysis mainly involved biological processes such as immune system response and cell proliferation,cellular components such as protein complex and cell junction,molecular functions such as transcription regulator activity and molecular function regulator;KEGG pathway enrichment mainly occurred in cancer-related pathways such as PI3K-Akt signaling pathway,Th17 cell differentiation,mTOR signaling pathway,FOXO signaling pathway,etc.The lncRNA-mediated ceRNA network was composed of 97 lncRNA,73 miRNA and 417 mRNA,and the circRNA-mediated ceRNA network was composed of 26 circRNA,68 miRNA and 157 mRNA.RT-qPCR validated the expressions of 15 differentially expressed genes,which was consistent with the sequencing results.Conclusion Cryoablation combined with Yangfei Prescription can inhibit the tumor growth of Lewis lung cancer bearing mice,destroy the structure of tumor tissue,inhibit cell proliferation,the mechanism may be related to the regulation of Gm38393/miRNA-136-5p/Zfp704 axis.

Objective:To evaluate the diagnostic value of coronary angiography-derived fractional flow reserve (FFR) and index of microcirculatory resistance (IMR) for identifying coronary functional abnormalities.Methods:This diagnostic study enrolled patients with clinically suspected or diagnosed coronary artery disease who underwent coronary angiography at Beijing Anzhen Hospital, TEDA International Cardiovascular Hospital, and Qilu Hospital of Shandong University between December 2021 and June 2022. All enrolled patients successfully underwent invasive wire-based FFR and IMR measurements during angiography. In a core laboratory, FFR and IMR for the target vessels were measured using artificial intelligence technology based on coronary angiographic images. Spearman correlation analysis was used to evaluate the correlation between angiography-derived FFR and wire-based FFR, and between angiography-derived IMR and wire-based IMR. Coronary hemodynamic abnormality was defined as FFR≤0.80; the diagnostic performance of angiography-derived FFR for identifying this abnormality was evaluated. Microcirculatory dysfunction was defined as IMR≥25; the diagnostic performance of angiography-derived IMR for identifying microcirculatory dysfunction was evaluated.Results:A total of 181 patients, aged (60.6±8.8) years, with 62 (34.3%) females, and 181 target vessels were included in the final analysis. Angiography-derived FFR showed a significant positive correlation with wire-based FFR ( r=0.78, P<0.001). For identifying coronary hemodynamic abnormality, angiography-derived FFR showed an accuracy of 89.0%, sensitivity of 88.8%, specificity of 89.1%, positive predictive value (PPV) of 88.8%, negative predictive value (NPV) of 89.1%, and an area under the receiver operating characteristic curve ( AUC) of 0.88. Angiography-derived IMR showed a significant positive correlation with wire-based IMR ( r=0.93, P<0.001). For identifying microcirculatory dysfunction, angiography-derived IMR demonstrated an accuracy of 89.5%, sensitivity of 86.8%, specificity of 90.2%, PPV of 70.2%, NPV of 96.3%, and an AUC of 0.95. Conclusion:Angiography-derived FFR and IMR exhibit strong correlations with their invasive wire-based counterparts and demonstrate high diagnostic value for assessing coronary hemodynamics and coronary microcirculatory function.

Objective:To summarize the clinical features, electroencephalogram (EEG) and magnetoencephalogram (MEG) of patients with pure paroxysmal kinesigenic dyskinesia (PKD) and PKD with epilepsy, so as to better distinguish them and guide the treatments.Methods:The clinical data of 200 patients diagnosed with PKD in the Outpatient Department of Xuanwu Hospital, Capital Medical University from 2000 to 2023 were analyzed retrospectively. The patients were divided into 2 groups: pure PKD (174 cases) and PKD with epilepsy (26 cases) according to whether accompanied by epilepsy. The differences in clinical features, drug therapy, EEG and MEG were compared between the 2 groups.Results:The clinical features of the 2 groups were essentially similar, and the proportion of PKD dyskinesia induced by emotional stress in the pure PKD group (54/174, 31.03%) was higher than that in the PKD with epilepsy group (2/26, 7.69%; χ 2=5.010, P=0.025). In terms of pharmacological response, carbamazepine was the most commonly used medication in both groups, but patients with PKD with epilepsy may need a higher therapeutic dosages (0.2-0.4 g/d, and gradually increased to 0.8 g/d) to effectively manage both dyskinesia and seizures. Regarding the EEG and MEG, the proportion of EEG abnormalities was higher in PKD patients with epilepsy, mainly manifested as focal spikes [1/70(1.43%) vs 9/21(42.86%), χ 2=24.268, P<0.001], together with aberrant MEG discharge (4/18 vs 3/5, χ 2=1.155, P=0.282). The MEG dipoles were mainly distributed in the brain regions close to the frontal lobe and central region. Conclusions:The clinical manifestations of motor symptoms of pure PKD and PKD with epilepsy are similar, and carbamazepine remains the most effective treatment. PKD patients with epilepsy have a higher proportion of abnormal EEG, mainly manifested as focal spikes, and are more likely to show abnormal discharge of MEG, which could be used to distinguish them.

Background and purpose:Epidermal growth factor receptor exon 20 T790M(EGFR T790M)mutation is one of the acquired resistance mechanisms in non-small cell lung cancer(NSCLC)against first-/second-generation EGFR tyrosine kinase inhibitors(EGFR TKIs).Additionally,EGFR T790M mutation can also be observed in NSCLC patients who have not undergone EGFR TKIs treatment.This study aimed to compare the clinical pathological characteristics and prognostic differences between NSCLC patients with de novo and acquired EGFR T790M mutation,and further explore the immune microenvironment features of acquired T790M mutation in NSCLC.Methods:This study retrospectively included 3 762 cases of NSCLC diagnosed at Fudan University Shanghai Cancer Center from April 2020 to September 2022.Among them,2 070 cases(55.02%)exhibited EGFR mutations,and 556 cases(14.77%)received EGFR TKIs treatment.Specifically,there were 119 cases(3.16%)of NSCLC with EGFR T790M mutation,including 51 cases(1.35%)of de novo T790M mutation and 68 cases(1.81%)of acquired EGFR T790M mutation.Clinical data of the patients were collected for comparative analysis between NSCLC patients with de novo and acquired T790M mutation.Multiple immunofluorescence histochemistry(mIHC)was employed to explore the immune microenvironment characteristics of NSCLC patients with acquired T790M mutation.Results:The proportion of de novo and acquired T790M mutations was higher in female patients compared to males.Patients with de novo T790M mutation tended to be younger.Both de novo and acquired T790M mutations were more commonly found in poorly differentiated carcinomas.Among NSCLC patients with de novo T790M mutation,there was a higher rate of programmed death ligand-1(PD-L1)expression(60.00%).In contrast,among NSCLC patients with acquired T790M mutation,the rate of PD-L1 expression was lower(22.39%).Acquired T790M mutation in NSCLC was often accompanied by TP53 alterations(39.7%).Cox regression analysis results indicated that mesenchymal to epithelial transition(MET)factor alteration was a risk factor for the occurrence of acquired T790M mutation(P=0.000 5).The average overall survival(OS)showed no significant difference between de novo and acquired T790M mutations(35.4 and 37.3 months respectively).However,patients with acquired T790M mutation exhibited a higher proportion of recurrence and metastasis.In acquired T790M mutation,there was a higher presence of immune cell infiltration within the stromal compartment,such as CD20+B cells,CD23+B cells,CD8+T cells,CD8+PD-1-/+cells,CD20+PD-1-/+cells and CD23+PD-1-/+cells.Additionally,the study found that when EGFR was accompanied by tumor suppressor gene(TSG)alterations,the average distance between tumor cells and CD8+T cells,CD20+B cells,CD8+PD-1+cells,CD20+PD-1+cells and CD23+PD-1+cells was closer compared to cases with only EGFR mutations.Conclusion:In comparison to patients with de novo T790M mutation,patients with acquired T790M mutation exhibit a lower rate of PD-L1 positivity.Acquired T790M mutation often accompanies TP53 alterations,and MET alteration is identified as a risk factor triggering acquired T790M mutation.Although patients with acquired T790M mutation face higher risk of recurrence and metastasis,their average OS does not significantly differ from those with de novo T790M mutation.In cases of acquired T790M mutation,the presence of TSG mutations can alter the spatial distribution of immune cells,potentially leading to benefits from immunotherapy.

Objective:To study the distribution and antibiotics resistance of the main pathogens of neonatal purulent meningitis in different regions of China.Methods:A retrospective descriptive clinical epidemiological study was conducted in children with neonatal purulent meningitis which admitted to 18 tertiary hospitals in different regions of China between January 2015 to December 2019. The test results of blood and cerebrospinal fluid, and drug sensitivity test results of the main pathogens were collected. The distributions of pathogenic bacteria in children with neonatal purulent meningitis in preterm and term infants, early and late onset infants, in Zhejiang Province and other regions outside Zhejiang Province, and in Wenzhou region and other regions of Zhejiang Province were analyzed. The chi-square test was used for statistical analysis.Results:A total of 210 neonatal purulent meningitis cases were collected. The common pathogens were Escherichia coli ( E. coli)(41.4%(87/210)) and Streptococcus agalactiae ( S. agalactiae)(27.1%(57/210)). The proportion of Gram-negative bacteria in preterm infants (77.6%(45/58)) with neonatal purulent meningitis was higher than that in term infants (47.4%(72/152)), and the difference was statistically significant ( χ2=15.54, P=0.001). There were no significant differences in the constituent ratios of E. coli (36.5%(31/85) vs 44.8%(56/125)) and S. agalactiae (24.7%(21/85) vs 28.8%(36/125)) between early onset and late onset cases (both P>0.05). The most common pathogen was E. coli in different regions, with 46.7%(64/137) in Zhejiang Province and 31.5%(23/73) in other regions outside Zhejiang Province. In Zhejiang Province, S. agalactiae was detected in 49 out of 137 cases (35.8%), which was significantly higher than other regions outside Zhejiang Province (11.0%(8/73)). The proportions of Klebsiella pneumoniae, and coagulase-negative Staphylococcus in other regions outside Zhejiang Province (17.8%(13/73) and 16.4%(12/73)) were both higher than those in Zhejiang Province (2.9%(4/137) and 5.1%(7/137)). The differences were all statistically significant ( χ2=14.82, 12.26 and 7.43, respectively, all P<0.05). The proportion of Gram-positive bacteria in Wenzhou City (60.8%(31/51)) was higher than that in other regions in Zhejiang Province (38.4%(33/86)), and the difference was statistically significant ( χ2=6.46, P=0.011). E. coli was sensitive to meropenem (0/45), and 74.4%(32/43) of them were resistant to ampicillin. E. coli had different degrees of resistance to other common cephalosporins, among which, cefotaxime had the highest resistance rate of 41.8%(23/55), followed by ceftriaxone (32.4%(23/71)). S. agalactiae was sensitive to penicillin, vancomycin and linezolid. Conclusions:The composition ratios of pathogenic bacteria of neonatal purulent meningitis are different in different regions of China. The most common pathogen is E. coli, which is sensitive to meropenem, while it has different degrees of resistance to other common cephalosporins, especially to cefotaxime.

Objective:To explore the establishment and application of ovarian cancer organoids.Methods:Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC 50) in different organoids. Results:(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC 50 of (29.5±15.8) μmol/L. Moreover, IC 50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022]. Conclusions:Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

Objective:To compare the detection and amplitude of epileptiform discharges (EDs) between surface sphenoidal electrode and anterior temporal electrode in patients with interictal EDs in the temporal region, and to explore the value of surface sphenoidal electrode.Methods:A total of 1 356 outpatients with epilepsy who underwent 2-hour video electroencephalogram (EEG) monitoring in Xuanwu Hospital from October to December 2021 were retrospectively enrolled. All patients were hooked up with scalp electrode according to the international 10-20 system as well as surface sphenoidal electrode and anterior temporal electrode. The EEGs with EDs recorded by surface sphenoidal electrode and/or anterior temporal electrode were selected for analysis. The detection rate and the amplitude of EDs by surface sphenoidal electrode and anterior temporal electrode were compared.Results:Seventy-three EEGs were collected and 250 EDs were counted. The detection rate of the anterior temporal electrode and surface sphenoidal electrode were 88.0% (220/250) and 98.4% (246/250) respectively. The difference in detection rate was statistically significant (χ 2=18.38, P<0.001). For the EDs from anterior temporal regions (taking the discharges recorded by anterior temporal electrode as "gold standard"), the detection rate of surface sphenoidal electrode was 98.2% (216/220). There was no statistically significant difference in detection rate between the anterior temporal electrode and surface sphenoidal electrode (χ 2=2.27, P=0.132). There were 216 EDs recorded by these two kinds of electrode simultaneously. The average amplitude of the EDs on surface sphenoidal electrode and anterior temporal electrode was (77.1±38.9) μV and (80.2±44.9) μV, respectively. The difference was statistically significant ( t=2.28, P=0.031). Conclusions:The detection rate of surface sphenoid electrodes was higher than that of anterior temporal electrodes for the EDs in the temporal region, and surface sphenoidal electrodes can be used routinely in outpatient. The surface sphenoidal electrode had more chance to detect EDs originating from regions out of the anterior temporal regions.

To summarize the nursing experience of an adolescent with early repolarization syndrome who had implanted a subcutaneous implantable cardioverter defibrillator(ICD)and experiences frequent ventricular fibrillation.The main nursing care were as follows:recognizing a ventricular fibrillation attack and emergency care;risk prevention and handling in transportation;prevention of iatrogenic infection and strengthening protective isolation;promoting the recovery of brain function;personalized health education;pre-hospital first aid skills training for families.After careful treatment and nursing,the patient did not recur arrhythmia,and heart function had improved significantly,and the pulmonary inflammation disappeared.No medical-acquired infections occurred during hospitalization.The patient's motor ability and language expression ability recovered to the baseline,and was discharged on the 20th day of admission.A month after discharge,the patient recovered well and could study and live normally.