Objective To construct large medical model named by "Huaxi HongYi"and explore its application effectiveness in assisting medical record generation. Methods By the way of a full-chain medical large model construction paradigm of "data annotation - model training - scenario incubation", through strategies such as multimodal data fusion, domain adaptation training, and localization of hardware adaptation, "Huaxi HongYi" with 72 billion parameters was constructed. Combined with technologies such as speech recognition, knowledge graphs, and reinforcement learning, an application system for assisting in the generation of medical records was developed. Results Taking the assisted generation of discharge records as an example, in the pilot department, after using the application system, the average completion times of writing a medical records shortened (21 min vs. 5 min) with efficiency increased by 3.2 time, the accuracy rate of the model output reached 92.4%. Conclusion It is feasible for medical institutions to build independently controllable medical large models and incubate various applications based on these models, providing a reference pathway for artificial intelligence development in similar institutions.

The comprehensive detection and identification of active ingredients in complex matrices is a crucial challenge.Liquid chromatography coupled with high-resolution mass spectrometry(LC-HRMS)is the most prominent analytical platform for the exploration of novel active compounds from complex matrices.However,the LC-HRMS-based analysis workflow suffers from several bottleneck issues,such as trace content of target compounds,limited acquisition for fragment information,and uncertainty in interpreting relevant MS2 spectra.Lycibarbarspermidines are vital antioxidant active ingredients in Lycii Fructus,while the reported structures are merely focused on dicaffeoylspermidines due to their low content.To comprehensively detect the new structures of lycibarbarspermidine derivatives,a"depict"strategy was developed in this study.First,potential new lycibarbarspermidine derivatives were designed according to the biosynthetic pathway,and a comprehensive database was established,which enlarged the coverage of lycibarbarspermidine derivatives.Second,the polarity-oriented sample prep-aration of potential new compounds increased the concentration of the target compounds.Third,the construction of the molecular network based on the fragmentation pathway of lycibarbarspermidine derivatives broadened the comprehensiveness of identification.Finally,the weak response signals were captured by data-dependent scanning(DDA)followed by parallel reaction monitoring(PRM),and the efficiency of acquiring MS2 fragment ions of target compounds was significantly improved.Based on the integrated strategy above,210 lycibarbarspermidine derivatives were detected and identified from Lycii Fructus,and in particular,170 potential new compounds were structurally characterized.The integrated strategy improved the sensitivity of detection and the coverage of low-response components,and it is expected to be a promising pipeline for discovering new compounds.

Objective: To establish a rapid and sensitive isothermal amplification assay for the detection of human Adenovirus. Methods: Primers and probe used for recombinase polymerase amplification(RPA)were designed based on the conserved region of the adenoviruses hexon gene. After optimizing the reaction temperature and times, the products of RPA were detected by capillary electrophoresis and lateral flow dipstick(LFD). Sensitivity and specicity of the assay were evaluated. The diagnostic value of the RPA-LFD assay was verified using clinical samples which were simultaneously tested by real time PCR assay. Results: The analytical sensitivity of RPA-LFD assay was 2 copies DNA molecules per reaction and no cross reaction with other pathogens was observed. Compared with real-time PCR assay, the sensitivity, and specificity of the present assay were all 100%. Conclusions The RPA-LFD assay developed in this study has the characteristics of high specificity, sensitivity, rapid and no requirement of expensive equipment which provided a new tool for rapid detection of human adenovirus.

Objective To establish a double-antibody sandwich ELISA for the rapid detection of shiga toxin typeⅡ ( StxⅡ) in shiga toxin-producing Escherichia coli ( STEC) infection. Methods A pool of murine hybridomas was used to screen out the optimal antibody pair for the establishment of double-anti-body sandwich ELISA. The established ELISA system was used to detect StxⅡin the culture supernatants of 16 clinical strains of STEC. Specificity and sensitivity of the established ELISA system were also evaluated. Results Two antibodies, S2D8 and S2C6, were successfully screened out, based on which the double-anti-body sandwich ELISA was set up. StxⅡand its variants rather than StxⅠwas detected in the culture super-natants of STEC with a lowest detection limit of 4 ng/ml. Its performance was consistent with that of commer-cial colloidal gold test kit, indicating the characteristics of good specificity and sensitivity. Conclusion The S2D8/S2C6-based ELISA laid a foundation for researches which designates the shiga toxin as a potential can-didate on the diagnosis and therapy of STEC infection.

OBJECTIVETo screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data.

METHODSGene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25% TBSA, full-thickness) database. After screening of the differentially expressed genes (DEGs) through paired-sample t-test and fold-change, DAVID online tools were used to select the DEGs related to leukocyte responses to burns by GO functional enrichment analysis; the interacting genes identified through KEGG pathway enrichment analysis were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR was used to verify the DEGs identified in mice.

RESULTSOf the 259 leukocyte response-related DEGs screened at 1 day post-burn, 118 were up-regulated and 141 were down-regulated. KEGG pathway enrichment analysis showed that the pathways were associated with the immune function, cell growth and cell death. PPI network and module analysis suggested that some of genes (such as Lck, Stat1, Myd88, Stat3, and Jun) play critical roles in the PPI network post-burn. RT-PCR results were consistent with those of bioinformatic analysis.

CONCLUSIONSLck, Stat1, Myd88, Stat3, and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several potential biomarkers for burn treatment.

Animals ; Burns ; genetics ; Computational Biology ; Down-Regulation ; Gene Expression Profiling ; Gene Regulatory Networks ; Leukocytes ; physiology ; Mice ; Real-Time Polymerase Chain Reaction ; Software ; Up-Regulation

Objective To evaluate the practicability of using CRISPR/Cas9 genome editing tech-nology for inhibition of hepatitis B virus ( HBV) replication. Methods Two sgRNA targeting sites were de-signed for the S region of HBV genome. The CRISPR/Cas9 expression plasmids specific for HBV were con-structed and then transfected into a cell line expressing HBV genome(HepG2-N10). The cytotoxicity of cells transfected with different expression plasmids were detected by MTT assay. The levels of hepatitis B surface antigen ( HBsAg ) were determined by using chemiluminescent immunoassay ( CLIA ) . The expression of HBV at mRNA level was analyzed by quantitative real-time PCR ( qRT-PCR) . The qPCR was performed for the detection of extracellular and intracellular HBV DNA. The next-generation sequencing ( NGS) Illumina MiSeq Platform was used to analyze HBV genome editing. Results No significant cytotoxic effects were de-tected in HepG2-N10 cells transfected with different expression plasmids. Compared with the cells carrying pCas-Guide-GFP-Scramble, the levels of HBsAg in the supernatants of transfected cell culture harboring pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were decreased by 24. 2% (P<0. 05) and 16. 9% (P>0. 05), respectively. The levels of HBsAg in cells transfected with pCas-Guide-GFP-G1 and pCas-Guide-GFP-G2 were respectively decreased by 16. 4% (P>0. 05) and 32. 1% (P>0. 05) as compared with that of pCas-Guide-GFP-Scramble transfected group. The expression of HBV at mRNA level was inhibited as indica-ted by the results of qRT-PCR. Moreover, the levels of extracellular HBV DNA were respectively suppressed by 23% (P>0. 05) and 35% (P<0. 05), and the levels of intracellular HBV DNA were respectively sup-pressed by 7. 2% (P>0. 05) and 18% (P>0. 05). Different types of insertion/deletion mutation were de-tected in HBV genome by high-throughput sequencing. Conclusion HBV-specific CRISPR/Cas9 system could inhibit the expression of HBV gene and the replication of virus. Therefore, the CRISPR/Cas9 genome editing technology might be used as a potential tool for the treatment of persistent HBV infection.

Objective To screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data. Methods Gene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25% TBSA, full-thickness) database. After screening of the differentially expressed genes (DEGs) through paired-sample t-test and fold-change, DAVID online tools were used to select the DEGs related to leukocyte responses to burns by GO functional enrichment analysis;the interacting genes identified through KEGG pathway enrichment analysis were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR was used to verify the DEGs identified in mice. Results Of the 259 leukocyte response-related DEGs screened at 1 day post-burn, 118 were up-regulated and 141 were down-regulated. KEGG pathway enrichment analysis showed that the pathways were associated with the immune function, cell growth and cell death. PPI network and module analysis suggested that some of genes (such as Lck, Stat1, Myd88, Stat3, and Jun) play critical roles in the PPI network post-burn. RT-PCR results were consistent with those of bioinformatic analysis. Conclusion Lck, Stat1, Myd88, Stat3, and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several potential biomarkers for burn treatment.

Objective To screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data. Methods Gene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25% TBSA, full-thickness) database. After screening of the differentially expressed genes (DEGs) through paired-sample t-test and fold-change, DAVID online tools were used to select the DEGs related to leukocyte responses to burns by GO functional enrichment analysis;the interacting genes identified through KEGG pathway enrichment analysis were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR was used to verify the DEGs identified in mice. Results Of the 259 leukocyte response-related DEGs screened at 1 day post-burn, 118 were up-regulated and 141 were down-regulated. KEGG pathway enrichment analysis showed that the pathways were associated with the immune function, cell growth and cell death. PPI network and module analysis suggested that some of genes (such as Lck, Stat1, Myd88, Stat3, and Jun) play critical roles in the PPI network post-burn. RT-PCR results were consistent with those of bioinformatic analysis. Conclusion Lck, Stat1, Myd88, Stat3, and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several potential biomarkers for burn treatment.

OBJECTIVETo investigate the effect of antibacterial peptide LL-37 on the integrity of Acinetobacter baumannii biofilm.

METHODSA model of Acinetobacter baumannii biofilm in vitro was constructed by plates and crystal violet staining method, and the minimal inhibitory concentration of LL-37 was determined by broth dilution. The biofilm morphology was observed by scanning electron microscopy and biofilm formation was analyzed by the crystal violet staining of the adherent biofilm in the presence of antibacterial peptide LL-37.

RESULTSIn the Acinetobacter baumannii biofilm model, the minimal inhibitory concentration of LL-37 was 64 µg/ml; LL-37 caused structural damage of the biofilm at a low concentration of 2.5 µg/ml. The biofilm was decreased gradually as the concentration of LL-37 increased.

CONCLUSIONLL-37 even at a concentration far below its minimal inhibitory concentration can cause structural damage of Acinetobacter baumannii biofilm in vitro.

Acinetobacter baumannii ; drug effects ; physiology ; Biofilms ; drug effects ; Cathelicidins ; pharmacology ; Microbial Sensitivity Tests

Objective To investigate the effect of antibacterial peptide LL-37 on the integrity of Acinetobacter baumannii biofilm. Methods A model of Acinetobacter baumannii biofilm in vitro was constructed by plates and crystal violet staining method, and the minimal inhibitory concentration of LL-37 was determined by broth dilution. The biofilm morphology was observed by scanning electron microscopy and biofilm formation was analyzed by the crystal violet staining of the adherent biofilm in the presence of antibacterial peptide LL-37. Results In the Acinetobacter baumannii biofilm model, the minimal inhibitory concentration of LL-37 was 64μg/ml;LL-37 caused structural damage of the biofilm at a low concentration of 2.5μg/ml. The biofilm was decreased gradually as the concentration of LL-37 increased. Conclusion LL-37 even at a concentration far below its minimal inhibitory concentration can cause structural damage of Acinetobacter baumannii biofilm in vitro.