Objective This study aims to investigate the therapeutic effect and mechanism of Naoxinqing on non-alcohol fatty liver disease (NAFLD) induced by a high-fat diet through network pharmacology,molecular docking and in vitro and in vivo experiments. Methods ApoE-/-mice were given a high-fat diet for 12 weeks to establish the NAFLD model,followed by a 12-week Naoxinqing administration. To evaluate the therapeutic effect of Naoxinqing on NAFLD induced by a high-fat diet,biochemical and histopathological experiments were performed,including assessment of blood lipids,liver function,serum inflammatory factors,as well as Hematoxylin and eosin (HE),Oil red O,and Sirius red staining of liver. Subsequently,network pharmacology and molecular docking techniques were employed to predict the key targets of Naoxinqing. Finally,the mechanism of Naoxinqing was validated by Western Blot in HepG2 cells and liver tissue. Results The results of serum biochemistry and liver tissue pathology showed that Naoxinqing can significantly improve high-fat diet-induced hepatic lipid accumulation,hepatocellular injury,and inflammation. Network pharmacology and molecular docking analysis results suggested that Naoxinqing may affect lipid metabolism through the AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway. Finally,in vitro cell experiment confirmed that the main mechanism of Naoxinqing is to activative the AMPK/SIRT1 pathway,upregulate the expression of downstream carnitine palmitoyltransferase 1 (CPT1A),promote fatty acid oxidation,and ultimately improve NAFLD. Conclusion This study demonstrated that Naoxinqing improved NAFLD by promoting fatty acid oxidation through the activation of the AMPK/SIRT1 pathway.

Objective To preliminary explore the in vitro anti-inflammatory effects of Qinggan Tongyin based on serum pharmacology and network pharmacology.Methods The effects of the serum containing Qinggan Tongyin on the release of NO,cell necrosis factor-α(TNF-α),and interleukin-6(IL-6)in LPS-induced RAW264.7 cells were confirmed using serum pharmacology.UHPLC-MS/MS was used to determine the index components of Qinggan Tongyin.The possible targets and pathways of active components in Qinggan Tongyin for anti-inflammatory properties were predicted by using network pharmacology.Results The results of cellular assay showed that Qinggan Tongyin could dramatically lessen the levels of NO,TNF-α,and IL-6(P＜0.05,P＜0.01,P＜0.001).The higher contents of Qinggan Tongyin were phillyrin A,arctiin,chlorogenic acid,scutellarin,gallic acid,rosmarinic acid,paeoniflorin and phillyrin.A totsl of 215 intersection targets between 17 active components in Qinggan Tongyin and inflammation were obtained,and the 31 core targets were ALB,VEGFA,IL-6,TNF-α,etc..The primary targets can exhibit anti-inflammatory actions by regulating several signaling pathways,such as AGE-RAGE,PI3K-Akt,and MAPK signaling pathway.Conclusion Qinggan Tongyin exerts its anti-inflammatory effects with the characteristic of multiple components and multiple targets.

Objective To investigate the application value of extracellular volume fraction(ECV)obtained from enhanced CT in diagnosis and classification of acute pancreatitis.Methods The clinical data from patients with acute pancreatitis(acute pancreatitis group)and normal controls(control group)underwent enhanced CT were analyzed retrospectively.The CT values of pancreas and abdominal aorta in the same sclice on precontrast and equilibrium-phase images were measured,and then pancreatic ECV was calcu-lated.The measured parameters were compared between the groups of control and acute pancreatitis,and subgroups of non-severe and severe pancreatitis.The logistic regression analysis was used to identify the risk factors for acute pancreatitis and severe pancrea-titis,and the receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficiency in diagnosis and classifica-tion of acute pancreatitis.Results The pancreatic CT value and ECV were independent risk factors for acute pancreatitis(P＜0.05),and the ECV was an independent risk factor for severe pancreatitis(P＜0.05).The area under the curve(AUC)of ECV was higher in acute pancreatitis group(0.81)and severe pancreatitis subgroup(0.68).Conclusion As a quantitative parameter,the ECV obtained from enhanced CT has higher clinical application value and higher popularity in the diagnosis and classification of acute pancreatitis.

This study aims to analyze the main drug resistance mechanism of Salmonella enteritidis induced by polymyxin in vitro.In this study,the resistance of Salmonella enteritidis CMCC(B)50335(ZK)to polymyxin was induced in vitro,and the growth characteristics,exercise ability,ul-trastructure and sensitivity to 16 antimicrobial agents before and after induction were determined by turbidimetry,semi-solid agar method,transmission electron microscope and disk diffusion method,and the whole genome and single nucleotide polymorphism(SNP)were detected by Illu-mina NovaSeq PE150 method.RT-qPCR was used to detect the differences in the expression levels of six drug-resistant related genes.The recombinant strain △phoPE1-128-1,which induced drug-resistant strain E1-128-1,was constructed by homologous recombination of Red Escherichia coli,and its sensitivity to polymyxin was detected.The results showed that three strains of induced drug-resistant bacteria E1-128-1,E1-128-3 and E2-128-3 were screened out,and the MIC increased by 128,64 and 64 times respectively after drug resistance stability test.Induced drug resistance had no significant effect on the growth ability of the tested bacteria and the sensitivity of 16 antibacte-rial drugs.The exercise ability of E1-128-1was significantly increased,and the cell wall and plasma membrane obviously thicken.There was no significant difference in the genome components of E1-128-1 before and after induction,but eight missense mutations of six drug-resistance related genes,including phoP/phoQ,cpxP,lptD,csrA and acrB,were detected,including four missense mutation sites of phoP,namely Leu185Trp,His189Ser,Thr190Tyr and Ile191His.The corresponding genes were sequenced by PCR,and the results were consistent with those of SNP.RT-qPCR results showed that the expression levels of mutant genes of the three induced strains increased signifi-cantly.Compared with E1-128-1,the MIC of △phoP E1-128-1 decreased to 1 mg/L.It is sug-gested that the mutation and increased expression of phoP gene are important factors for inducing polymyxin resistance of Salmonella CMCC(B)50335 in vitro.

Objective To investigate the toxicity and mechanism of a novel Staphylococcus aureus with incomplete hemolytic phenotype(SIHP)filtrate on non-small cell lung cancer(NSCLC)HCC827 cell line.Methods The filtrates and diluents of novel SIHP and Staphylococcus aureus with complete hemolytic phenotype(SCHP)were co-incubated with 1％ human red blood cells and HCC827 cells.Hemoglobin release method was used to detect red blood cell toxicity,CCK-8 method was used to detect HCC827 cell activity.Ion selective electrode method,colori-metric method and immunofluorescence method were used to detect potassium(K+),lactate dehydrogenase(LDH)and interleukin-6(IL-6)concentrations in the incubation supernatants.Results The filtrate and diluent of novel SIHP could significantly damage the activity of HCC827 cells,causing K+and LDH leakage.The filtrate and 1:3 diluent of novel SIHP resulted in a decrease and an increase in IL-6 secretion in HCC827 cells,respec-tively.Compared with SCHP,the filtrate and diluent of novel SIHP had stronger red blood cell toxicity and more severe K+leakage in HCC827 cells.Conclusion The filtrate of novel SIHP can damage the cell membrane,lead to content release and kill NSCLC cells,with a stronger toxicity than SCHP.

Background::Breast cancer (BC) risk-stratification tools for Asian women that are highly accurate and can provide improved interpretation ability are lacking. We aimed to develop risk-stratification models to predict long- and short-term BC risk among Chinese women and to simultaneously rank potential non-experimental risk factors.Methods::The Breast Cancer Cohort Study in Chinese Women, a large ongoing prospective dynamic cohort study, includes 122,058 women aged 25-70 years old from the eastern part of China. We developed multiple machine-learning risk prediction models using parametric models (penalized logistic regression, bootstrap, and ensemble learning), which were the short-term ensemble penalized logistic regression (EPLR) risk prediction model and the ensemble penalized long-term (EPLT) risk prediction model to estimate BC risk. The models were assessed based on calibration and discrimination, and following this assessment, they were externally validated in new study participants from 2017 to 2020.Results::The AUC values of the short-term EPLR risk prediction model were 0.800 for the internal validation and 0.751 for the external validation set. For the long-term EPLT risk prediction model, the area under the receiver operating characteristic curve was 0.692 and 0.760 in internal and external validations, respectively. The net reclassification improvement index of the EPLT relative to the Gail and the Han Chinese Breast Cancer Prediction Model (HCBCP) models for external validation was 0.193 and 0.233, respectively, indicating that the EPLT model has higher classification accuracy.Conclusions::We developed the EPLR and EPLT models to screen populations with a high risk of developing BC. These can serve as useful tools to aid in risk-stratified screening and BC prevention.

OBJECTIVE: To explore the clinical phenotype and genetic basis of a child with early onset neurodevelopmental disorder with involuntary movement (NEDIM). METHODS: A child who presented at Department of Neurology of Hunan Children's Hospital on October 8, 2020 was selected as the study subject. Clinical data of the child were collected. Genomic DNA was extracted from peripheral blood samples of the child and his parents. Whole exome sequencing (WES) was carried out for the child. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. Relevant literature was searched from the CNKI, PubMed and Google Scholar databases to summarize the clinical phenotypes and genetic variants of the patients. RESULTS: This child was a 3-year-and-3-month boy with involuntary trembling of limbs and motor and language delay. WES revealed that the child has harbored a c.626G>A (p.Arg209His) variant of the GNAO1 gene. Sanger sequencing confirmed that neither of his parents has carried the same variant. The variant had been reported in HGMD and ClinVar databases, but not in the dbSNP, ExAC and 1000 Genomes databases. Prediction with SIFT, PolyPhen-2, and Mutation Taster online software suggested that the variant may be deleterious to the protein function. By UniProt database analysis, the encode amino acid is highly conserved among various species. Prediction with Modeller and PyMOL software indicated that the variant may affect the function of GαO protein. Based on the guideline of the American College of Medical Genetics and Genomics (ACMG), the variant was rated as pathogenic. CONCLUSION The GNAO1 gene c.626G>A (p.Arg209His) variant probably underlay the NEDIM in this child. Above finding has expanded the phenotypic spectrum of GNAO1 gene c.626G>A (p.Arg209His) variant and provided a reference for clinical diagnosis and genetic counseling.
Humans ; Computational Biology ; Genetic Counseling ; Genomics ; Mutation ; Neurodevelopmental Disorders/genetics* ; Dyskinesias ; GTP-Binding Protein alpha Subunits, Gi-Go

Objective:To investigate the relationship between carotid atherosclerosis(CAS)and subclinical left ventricular(LV)dysfunction in type 2 diabetes mellitus patients with preserved LV ejection fraction(LVEF).Methods:A total of 120 patients with type 2 diabetes mellitus who had LVEF≥50% were selected in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University from June 2021 to October 2021. The global longitudinal strain(GLS)was obtained by two-dimensional speckle tracking echocardiography(STE)to assess subclinical LV systolic function. The mitral ratio of peak early to late diastolic filling velocity(E/A), and mitral velocity to early diastolic velocity of the mitral annulus(E/E′)ratio were obtained by pulsed tissue Doppler echocardiography to assess LV diastolic function. Acrroding to bilateral carotid ultrasound examination, the subjects were divided into normal carotid arteries group( n=46) and CAS group( n=74). Demographics and biochemical parameters were compared between two groups. Binary logistic regression and Pearson correlation analysis were used to evaluate the relationship between CAS and subclinical LV dysfunction. Results:The CAS group had a higher proportion of men, older age, and a longer duration of diabetes than the normal carotid arteries group(all P<0.05). There was no difference in LVEF and GLS between the two groups [normal carotid arteries group vs CAS group, LVEF: (60.72±4.73)% vs(60.07±4.28)%; GLS: (18.24±3.72)% vs(17.81±3.47)%, respectively; both P>0.05]. However, compared with normal carotid arteries group, E/A ratio was decreased and E/E′ ratio was significantly increased in CAS group(both P<0.01). Pearson correlation analysis showed that GLS was not correlated with carotid plaque thickness and carotid intima-media thickness(CIMT; both P>0.05). By contrast, E/E′ ratio was positively correlated with carotid plaque thickness and CIMT(both P<0.05). Binary logistic regression analysis showed that GLS and E/E′ ratio were not associated with CAS( both P>0.05). However, decreased E/A ratio was significantly associated with the existence of CAS( OR=0.09, 95% CI 0.01-0.67, P=0.018). Conclusions:In type 2 diabetes mellitus patients without overt heart failure and with preserved LVEF, the occurrence of CAS is not associated with subclinical LV systolic impairment assessed by GLS, but is significantly associated with LV diastolic dysfunction, and is independent of traditional cardiovascular risk factors.

Objective:To establish the high performance liquid chromatography (HPLC)fingerprints of Zhuanggu Guanjie Pills, which provided the basis for its quality evaluation.Methods:HPLC was used with Agilent Eclipse XDB-C 18 column (4.6 mm×250 mm, 5 μm);mobile phase was acetonitrile-0.1% acetic acid water; gradient elution; flow rate was 1 ml/min; column temperature was at 35 ℃; detection wavelength was 254 nm; injection volume was 10 μl. The HPLC fingerprints of 15 batches of Zhuanggu Guanjie Pills were established and similarity analysis was carried out, and the contents of 18 components were estimated. Results:In the fingerprint study, isopsoralen was used as the reference peak, 40 common peaks were marked and 18 peaks were identified and the similarity between the fingerprints of 15 batches of Zhuanggu Guanjie Pills and the control fingerprints was greater than 0.99.Conclusion:This method is easy to operate and has high accuracy, which can provide basis and reference for the quality evaluation of Zhuanggu Guanjie Pills.

Objective : To investigate the effects of dihydroartemisinin ( DHA) on cell growth , migration invasion and vasculogenic mimicry ability of non⁃small cell lung cancer ( NSCLC) . Methods : Different concentrations of DHA were added to NSCLC cell lines A549 and H3255 , and after 24 hours , the cell viability was detected by CCK8 method , and the migration and invasion ability of NSCLC cells was detected by Transwell experiment . qRT⁃PCR and Western blot detected E ⁃cadherin , N ⁃cadherin , and Vimentin mRNA and protein expression levels , respectively .Three⁃dimensional cell was cultured to observe the vascular⁃like morphological generation of cells . qRT⁃PCR and Western blot detected human vascular endothelial cadherin (VE⁃cadherin) mRNA and protein expression levels as marker of vasculogenic mimicry . Results : DHA inhibited cell growth of A549 and H3255 and showed time⁃dependent and concentration⁃dependent . DHA inhibits the invasion and metastasis ( all P < 0. 001) of A549 and H325 cells ; DHA upregulated the expression of E ⁃Cadherin at mRNA(all P < 0. 001) and protein(P < 0. 001;P < 0. 01) levels in A549 and H3255 cells , and downregulated the expression of N ⁃cadherin at mRNA (all P < 0. 01) and protein (all P < 0. 001) levels as well as the expression of Vimentin at mRNA (P < 0. 01;P < 0. 001) and protein (all P < 0. 001) levels . The results of three⁃dimensional cell culture showed that the 12⁃hour vasculogenic mimicry generation capacity of DHA⁃treated A549 cells and H3255 cells was reduced , and the expressions of VE⁃cadherin at mRNA (P < 0. 001;P < 0. 01) and protein (all P < 0. 001) levels were downregulated . Conclusion Dihydroartemisinin inhibits the growth , migration invasion and vasculogenic mimicry ability of non⁃small cell lung cancer cells