BACKGROUND: Postoperative pulmonary complications (PPCs) are major adverse events in neurosurgical patients. This study aimed to develop and validate machine learning models predicting PPCs after neurosurgery. METHODS: PPCs were defined according to the European Perioperative Clinical Outcome standards as occurring within 7 postoperative days. Data of cases meeting inclusion/exclusion criteria were extracted from the anesthesia information management system to create three datasets: The development (data of Huashan Hospital, Fudan University from 2018 to 2020), temporal validation (data of Huashan Hospital, Fudan University in 2021) and external validation (data of other three hospitals in 2023) datasets. Machine learning models of six algorithms were trained using either 35 retrievable and plausible features or the 11 features selected by Lasso regression. Temporal validation was conducted for all models and the 11-feature models were also externally validated. Independent risk factors were identified and feature importance in top models was analyzed. RESULTS: PPCs occurred in 712 of 7533 (9.5%), 258 of 2824 (9.1%), and 207 of 2300 (9.0%) patients in the development, temporal validation and external validation datasets, respectively. During cross-validation training, all models except Bayes demonstrated good discrimination with an area under the receiver operating characteristic curve (AUC) of 0.840. In temporal validation of full-feature models, deep neural network (DNN) performed the best with an AUC of 0.835 (95% confidence interval [CI]: 0.805-0.858) and a Brier score of 0.069, followed by Logistic regression (LR), random forest and XGBoost. The 11-feature models performed comparable to full-feature models with very close but statistically significantly lower AUCs, with the top models of DNN and LR in temporal and external validations. An 11-feature nomogram was drawn based on the LR algorithm and it outperformed the minimally modified Assess respiratory RIsk in Surgical patients in CATalonia (ARISCAT) and Laparoscopic Surgery Video Educational Guidelines (LAS VEGAS) scores with a higher AUC (LR: 0.824, ARISCAT: 0.672, LAS: 0.663). Independent risk factors based on multivariate LR mostly overlapped with Lasso-selected features, but lacked consistency with the important features using the Shapley additive explanation (SHAP) method of the LR model. CONCLUSIONS: The developed models, especially the DNN model and the nomogram, had good discrimination and calibration, and could be used for predicting PPCs in neurosurgical patients. The establishment of machine learning models and the ascertainment of risk factors might assist clinical decision support for improving surgical outcomes. TRIAL REGISTRATION ChiCTR 2100047474; https://www.chictr.org.cn/showproj.html?proj=128279 .
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Algorithms ; Lung Diseases/etiology* ; Machine Learning ; Neurosurgical Procedures/adverse effects* ; Postoperative Complications/diagnosis* ; Risk Factors ; ROC Curve

Objective Five chloroplast(cp)genomes from members of genus Polygonatum were assembled by hybrid assembly technique,and their intraspecic and interspecific differences were analyzed by comparative genomic method.Codon usage patterns and influencing factors were determined,and the cp genome data were applied to understand the phylogenomic relationships in the entire genus Polygonatum along with the available data.Methods In this study,the chloroplast genomes of 5 species of genus Polygonatum were assembled using Unicycler software.Sequence alignment,collinearity analysis,boundary analysis and other methods were used to evaluate the interspecific differences of these five species.Nucleotide polymorphism analysis was used to discover the high-variation sites of the five species and their related species,predict the distribution of different long repeat sequences and SSRs,and then analyze the use bias of Polygonatum code.Finally,phylogenetic tree was constructed with the coding sequences of other 47 genus Polygonatum and their closely related species to explore their phylogenetic relationships in this study.Results ①Chloroplast genomes of 155 408-155 623 bp were assembled from five species of Polygonatum.A total of 132-133 genes were annotated,and 369 long repeats and 1553 simple repeats were detected.②The contraction and expansion of chloroplast genomes in 8 species were not obvious at the IRs boundary,and the size and distribution of individual genes at the LSC-IRs-SSC boundary,such as ndhF gene and ycf1 gene,were slightly different.No interspecific or intraspecific rearrangement was observed in 8 species.③ The high-variation regions of the 8 chloroplast genomes are mainly located in two single-copy regions,the duplicate copy region is relatively conserved,and the coding region is more conserved than the non-coding region.High nucleotide polymorphic loci rps16-trnQ,trnS-trnG,trnTUGU-trnL,ndhF-rpl32 and rpl32-trnL are located in the single copy region and most of them are gene spacer regions.④ The codon preference results showed that the codon preference of the five species was similar and mainly affected by selection pressure,and the third base of the codon played A dominant role and mainly ended in A/U.RSCU clustering heat map shows that PK and PZ,PCB and PS have close relationship.⑤ Phylogenetic trees divided 52 species into five branches:Ⅰ,Ⅱ,Ⅲ,ⅣandⅤ.PS,PK,PCB,PCZ and PZ were divided into ⅣandⅤbranches,among which PK and PZ were most closely related,while PCZ was more distant than the other four,was divided into the Ⅴbranch alone.Conclusion This study provided a reference for the phylogenetic research and molecular marker development of the medicinal plants of the Polygonum genus.

OBJECTIVE To investigate the activity and mechanism of tetrandrine(TET)against influenza A virus in vitro and in vivo.METHODS(1)Cell experiments.① Human non-small cell lung cancer cells(A549)were divided into TET 0(cell control),1.25,2.5,5,10,20 and 25 μmol·L-1 groups,and H1N1+TET 0,1.25,2.5,5,10,20 and 25 μmol·L-1 groups.The TET groups were treated with the corresponding concentrations of TET while the H1N1+TET groups were infected with H1N1 for 1 h before the corresponding concentrations of TET were added.After 48 h,cell viability was detected using the CCK-8 method.② The cells were divided into cell control,H1N1+TET 0,2.5,5,and 10 μmol·L-1 groups and treated as in ①.After 24 h of incubation,the mRNA expressions of matrix protein 1(M1),hemagglutinin(HA),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-β(IFN-β)were tested by the real-time quantitative PCR(RT-qPCR).The expression levels of M1,HA,neuraminidase(NA),nucleoprotein(NP),and phosphorylation of signal transducer and activator of transcription 3(STAT3)protein were detected by Western blotting.(2)Animal experiments.① Male BALB/c mice were randomly divided into the solvent control group,H1N1 group,H1N1+oseltamivir phosphate(Ose)20 mg·kg-1 group,and H1N1+TET 25,50 and 100 mg·kg-1 groups.The solvent control group and the H1N1 group were ig administered with 0.5％carboxymethyl cellulose sodium(CMC-Na),while the H1N1+Ose group and the H1N1+TET 25,50 and 100 mg·kg-1 groups were ig given suspensions of the respective concentrations of drugs in 0.5％CMC-Na.After three consecutive days of pretreatment,all these groups except the solvent control group were intranasally inoculated with H1N1 to establish an influenza-infected mouse model.The survival rate and body mass of mice were monitored and recorded for 15 consecutive days post-H1N1 infection.② The grouping and treatment were the same as ①.After infection,mice were sacrificed on day 3 and 5.The expression levels of M1,HA,TNF-α,IL-1βand IL-6in lung tissues were detected by RT-qPCR,and those of M1,HA,NA,NP,and phosphoryla-tion of STAT3 protein in mice lung tissues by Western blotting.Hematoxylin-Eosin(HE)staining was performed to observe the pathological changes of lung tissues in mice.The levels of IL-6,TNF-α and IFN-β in bronchoalveolar lavage fluid(BALF)were determined by enzyme-linked immunosorbent assays(ELISA).RESULTS(1)① The half maximal inhibitory concentration study showed a value of 18.06 μmol·L-1 for A549 effected by TET.Compared with the H1N1 group,TET 2.5,5 and 10 μmol·L-1 significantly increased cell viability.② The expression levels of M1,HA mRNA and M1,HA,NA protein in the TET 2.5,5 and 10 μmol·L-1 groups were significantly lowered compared with the H1N1 group.TET 5 μmol·L-1 significantly decreased H1N1-induced IL-6,TNF-α and IFN-β mRNA expression levels in A549 cells.TET 5 and 10 μmol·L-1 could significantly mitigate the phosphorylation of STAT3.(2)① Com-pared with the H1N1 group,TET 50 mg·kg-1 significantly improved the survival rate of H1N1-infected mice while TET 25 mg·kg-1 significantly elevated the body-weight of H1N1-infected mice.In the TET 50 mg·kg-1 group,expressions of HA and M1 mRNA,and HA,M1,NA and NP protein in the lung tissues of H1 N1-infected mice were significantly reduced compared with the H1N1 group.Compared with the H1N1 group,TET 50 mg·kg-1 significantly decreased the lung index,improved inflammatory lesions in lung tissues,inhibited the mRNA expressions of TNF-α,IL-6 and IFN-β in lung tissues,and down regu-lated the expressions of TNF-α,IL-6 and IFN-β proinflammatory cytokines in the BALF of the H1N1-infected mice.In addition,TET 50 mg·kg-1 also significantly inhibited STAT3 phosphorylation in lung tissues of mice infected with H1N1.CONCLUSION TET can inhibit H1N1 infection both in vivo and in vitro.The potential mechanism may be related to the inhibition of the IL-6/STAT3 pathway,which subse-quently suppresses the inflammatory response induced by H1N1.

OBJECTIVE To investigate the activity and mechanism of tetrandrine(TET)against influenza A virus in vitro and in vivo.METHODS(1)Cell experiments.① Human non-small cell lung cancer cells(A549)were divided into TET 0(cell control),1.25,2.5,5,10,20 and 25 μmol·L-1 groups,and H1N1+TET 0,1.25,2.5,5,10,20 and 25 μmol·L-1 groups.The TET groups were treated with the corresponding concentrations of TET while the H1N1+TET groups were infected with H1N1 for 1 h before the corresponding concentrations of TET were added.After 48 h,cell viability was detected using the CCK-8 method.② The cells were divided into cell control,H1N1+TET 0,2.5,5,and 10 μmol·L-1 groups and treated as in ①.After 24 h of incubation,the mRNA expressions of matrix protein 1(M1),hemagglutinin(HA),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interferon-β(IFN-β)were tested by the real-time quantitative PCR(RT-qPCR).The expression levels of M1,HA,neuraminidase(NA),nucleoprotein(NP),and phosphorylation of signal transducer and activator of transcription 3(STAT3)protein were detected by Western blotting.(2)Animal experiments.① Male BALB/c mice were randomly divided into the solvent control group,H1N1 group,H1N1+oseltamivir phosphate(Ose)20 mg·kg-1 group,and H1N1+TET 25,50 and 100 mg·kg-1 groups.The solvent control group and the H1N1 group were ig administered with 0.5％carboxymethyl cellulose sodium(CMC-Na),while the H1N1+Ose group and the H1N1+TET 25,50 and 100 mg·kg-1 groups were ig given suspensions of the respective concentrations of drugs in 0.5％CMC-Na.After three consecutive days of pretreatment,all these groups except the solvent control group were intranasally inoculated with H1N1 to establish an influenza-infected mouse model.The survival rate and body mass of mice were monitored and recorded for 15 consecutive days post-H1N1 infection.② The grouping and treatment were the same as ①.After infection,mice were sacrificed on day 3 and 5.The expression levels of M1,HA,TNF-α,IL-1βand IL-6in lung tissues were detected by RT-qPCR,and those of M1,HA,NA,NP,and phosphoryla-tion of STAT3 protein in mice lung tissues by Western blotting.Hematoxylin-Eosin(HE)staining was performed to observe the pathological changes of lung tissues in mice.The levels of IL-6,TNF-α and IFN-β in bronchoalveolar lavage fluid(BALF)were determined by enzyme-linked immunosorbent assays(ELISA).RESULTS(1)① The half maximal inhibitory concentration study showed a value of 18.06 μmol·L-1 for A549 effected by TET.Compared with the H1N1 group,TET 2.5,5 and 10 μmol·L-1 significantly increased cell viability.② The expression levels of M1,HA mRNA and M1,HA,NA protein in the TET 2.5,5 and 10 μmol·L-1 groups were significantly lowered compared with the H1N1 group.TET 5 μmol·L-1 significantly decreased H1N1-induced IL-6,TNF-α and IFN-β mRNA expression levels in A549 cells.TET 5 and 10 μmol·L-1 could significantly mitigate the phosphorylation of STAT3.(2)① Com-pared with the H1N1 group,TET 50 mg·kg-1 significantly improved the survival rate of H1N1-infected mice while TET 25 mg·kg-1 significantly elevated the body-weight of H1N1-infected mice.In the TET 50 mg·kg-1 group,expressions of HA and M1 mRNA,and HA,M1,NA and NP protein in the lung tissues of H1 N1-infected mice were significantly reduced compared with the H1N1 group.Compared with the H1N1 group,TET 50 mg·kg-1 significantly decreased the lung index,improved inflammatory lesions in lung tissues,inhibited the mRNA expressions of TNF-α,IL-6 and IFN-β in lung tissues,and down regu-lated the expressions of TNF-α,IL-6 and IFN-β proinflammatory cytokines in the BALF of the H1N1-infected mice.In addition,TET 50 mg·kg-1 also significantly inhibited STAT3 phosphorylation in lung tissues of mice infected with H1N1.CONCLUSION TET can inhibit H1N1 infection both in vivo and in vitro.The potential mechanism may be related to the inhibition of the IL-6/STAT3 pathway,which subse-quently suppresses the inflammatory response induced by H1N1.

Objective Five chloroplast(cp)genomes from members of genus Polygonatum were assembled by hybrid assembly technique,and their intraspecic and interspecific differences were analyzed by comparative genomic method.Codon usage patterns and influencing factors were determined,and the cp genome data were applied to understand the phylogenomic relationships in the entire genus Polygonatum along with the available data.Methods In this study,the chloroplast genomes of 5 species of genus Polygonatum were assembled using Unicycler software.Sequence alignment,collinearity analysis,boundary analysis and other methods were used to evaluate the interspecific differences of these five species.Nucleotide polymorphism analysis was used to discover the high-variation sites of the five species and their related species,predict the distribution of different long repeat sequences and SSRs,and then analyze the use bias of Polygonatum code.Finally,phylogenetic tree was constructed with the coding sequences of other 47 genus Polygonatum and their closely related species to explore their phylogenetic relationships in this study.Results ①Chloroplast genomes of 155 408-155 623 bp were assembled from five species of Polygonatum.A total of 132-133 genes were annotated,and 369 long repeats and 1553 simple repeats were detected.②The contraction and expansion of chloroplast genomes in 8 species were not obvious at the IRs boundary,and the size and distribution of individual genes at the LSC-IRs-SSC boundary,such as ndhF gene and ycf1 gene,were slightly different.No interspecific or intraspecific rearrangement was observed in 8 species.③ The high-variation regions of the 8 chloroplast genomes are mainly located in two single-copy regions,the duplicate copy region is relatively conserved,and the coding region is more conserved than the non-coding region.High nucleotide polymorphic loci rps16-trnQ,trnS-trnG,trnTUGU-trnL,ndhF-rpl32 and rpl32-trnL are located in the single copy region and most of them are gene spacer regions.④ The codon preference results showed that the codon preference of the five species was similar and mainly affected by selection pressure,and the third base of the codon played A dominant role and mainly ended in A/U.RSCU clustering heat map shows that PK and PZ,PCB and PS have close relationship.⑤ Phylogenetic trees divided 52 species into five branches:Ⅰ,Ⅱ,Ⅲ,ⅣandⅤ.PS,PK,PCB,PCZ and PZ were divided into ⅣandⅤbranches,among which PK and PZ were most closely related,while PCZ was more distant than the other four,was divided into the Ⅴbranch alone.Conclusion This study provided a reference for the phylogenetic research and molecular marker development of the medicinal plants of the Polygonum genus.

Objective: To analyze the incidence and mutation characteristics of FLT3 gene mutation and clinical efficacy of tyrosine kinase inhibitor (TKI) in patients with mixed phenotype acute leukemia (MPAL). Methods: A total of 48 patients with MPAL who were admitted to Hebei Yanda Lu Daopei Hospital from June 2015 to February 2018 were retrospectively analyzed. The common mutated 58 genes in hematologic malignancies were detected by using amplicon-targeted next generation sequencing, of which internal tandem duplication (ITD) and point mutation occurred in the hotspot region of exon 14, 15 and 20 in FLT3 gene. Multiplex polymerase chain reaction (PCR) analysis was used to detect 35 gene fusions in hematological neoplams. Results: There were 7 cases of FLT3 mutation in 48 MPAL patients, which were all ITD mutations. The median length of the inserts of FLT3-ITD was 48 bp, and one MPAL patient carried 2 multiple length inserts simultaneously, and the median variant allele frequency (VAF) was 40.5% (7.9%-84.7%). There were no statistically significant differences in clinical and genetic characteristics between FLT3 mutation-positive and FLT3 mutation-negative MPAL patients (both P > 0.05). Among 7 FLT3 mutation-positive MPAL patients, 4 cases were often accompanied with RUNX1 mutation. A total of 4 MPAL patients with FLT3-ITD-positive received sorafenib or sunitinib combined chemotherapy, and 3 of them achieved complete remission. Conclusions ITD mutation is the main part in the FLT3 mutation of MPAL patients. FLT3-ITD-positive MPAL patients are often accompanied with RUNX1 mutation, which may benefit from targeted therapy with FLT3 kinase inhibitor.

This retrospective analysis aimed to investigate the mutation profile of 16 common mutated genes in de novo acute myeloid leukemia (AML) patients. A total of 259 patients who were diagnosed of de novo AML were enrolled in this study. Mutation profiling of 16 candidate genes were performed in bone marrow samples by using Sanger sequencing.We identified at least 1 mutation in 199 of the 259 samples (76.8%), and 2 or more mutations in 31.7% of samples. FLT3-ITD was the most common mutated gene (16.2%, 42/259), followed by CEBPA (15.1%, 39/259), NRAS (14.7%, 38/259), and NPM1 (13.5%, 35/259). Concurrence was observed in 97.1% of the NPM1 mutated cases and in 29.6% of the double mutated CEBPA cases. Distinct patterns of co-occurrence were observed for different hotspot mutations within the IDH2 gene: R140 mutations were associated with NPM1 and/or FLT3-ITD mutations, whereas R172 mutations co-occurred with DNMT3A mutations only. Concurrence was also observed in 86.6% of epigenetic regulation genes, most of which co-occurred with NPM1 mutations. The results showed certain rules in the mutation profiling and concurrence of AML patients, which was related to the function classification of genes. Defining the mutation spectrum and mutation pattern of AML will contribute to the comprehensive assessment of patients and identification of new therapeutic targets.
Adolescent ; Adult ; Aged ; CCAAT-Enhancer-Binding Proteins ; genetics ; Child ; Child, Preschool ; China ; DNA Mutational Analysis ; Female ; GTP Phosphohydrolases ; genetics ; Gene Expression Profiling ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Kaplan-Meier Estimate ; Leukemia, Myeloid, Acute ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Phenotype ; Retrospective Studies ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

Obgective To observe the effect of Milrinone combined with Esmolol in the treatment of severe hand-foot-and-mouth disease (HFMD) so as to improve the prognosis.Methods Eighty-two cases of children with critically severe HFMD,who were hospitalized in the Intensive Care Unit of Xuzhou Children's Hospital,were enrolled in the study from may of 2013 to June of 2014,and were randomly divided into a control group and an observation group.The control group was given intravenous Milrinone,and the observation group was given Milrinone combined with Esmolol.The heart rate (HR),systolic blood pressure (SBP),cardiac output (CO),left ventricular ejection fraction (LVEF) and brain natriuretic peptide (BNP),norepinephrine (NE) were detected on admission and checked again 1 hour and 48 hours again after treatment.The changes in the above indicators were compared before and after therapy to evaluate the clinical curative effect.Results (1) There was no significant difference in the HR,SBP,CO,LVEF,BNP and NE between the 2 groups before treatment(all P ＞ 0.05).(2) The HR,SBP,CO and LVEF of 2 groups were significantly improved after 1 hour treatment compared with those before treatment (all P ＜ 0.05),and the BNP and NE of the control group were not obviously improved compared with those before therapy (all P ＞ 0.05),but the significant changes were seen in the observation group (all P ＜ 0.05).Forty-eight hours after the treatment,all the observed indicators in 2 groups were significantly improved compared with those before treatment(all P ＜ 0.01).(3)Compared with control group,the HR,SBP,CO,LVEF and BNP of the observation group were significantly improved after 1 hour treatment (t =2.08,2.12,-2.11,-2.37,2.07,all P ＜ 0.05),but the NE of the observation group had no obvious improvement (t =0.83,P ＞ 0.05).All the observed indicators of the observation group were significantly improved compared with the control group after 48 hours treatment(t =3.76,2.48,-2.70,-2.27,5.37,2.74,all P ＜ 0.05).Conclusions Milrinone combined with Esmolol can significantly improve the cardiac function and the vital signs of the children with critically severe HFMD,which can be recommended clinically.

Objective: To investigate the relation between proliferation, apoptosis and Bcl-2 in human tongue squamous cell carcinoma(TSCC). Methods: Apoptosis, prolifexation and Bcl-2 protein expression were examined in 7 cases of normal tongue mucosa, 20(10 of Han and 10 of Uygur people) of tongue squamous cell papilloma (TSCP) and 42 of TSCC (30 of well-differentiated and 12 of middle-differentiated ) with immunohistochemical and in situ cell death detection technique. Results: The apoptosis index (TI) and the proliferation index (PI) showed no significant difference between Han and Uygur people or between male and femae, TI and PI in human TSCP were heigher than those in normal tongue mucosa, The proliferation was enhanced and apoptosis was inhibited according to dysplasia degree(P