ObjectivePeripheral nerve injury is a common clinical condition that can impair the regulatory function of target organs, leading to functional disability; severe injuries may even result in lifelong disability. Therefore, how to effectively promote the morphological and functional reconstruction of injured peripheral nerves has become a critical scientific issue that urgently needs to be addressed. This study biomimicked the highly oriented structure and electrophysiological properties of peripheral nerves and investigated the feasibility and effectiveness of combining electrical stimulation (ES) with oriented conductive substrates for the repair of injured peripheral nerves at the cellular level. MethodsPolydimethylsiloxane (PDMS) substrates with oriented microgroove structures of varying widths (0, 5, 10, and 20 μm) were fabricated using silicon wafers with micropatterned grooves as templates. Graphene-based conductive patterned substrates (GCPSs) with oriented microgrooves were then constructed via soft lithography and designated as GCPS 0, GCPS 5, GCPS 10, and GCPS 20 according to the groove width. The GCPSs were prepared from collagen/polycaprolactone (PCL) matrices containing 1.0 wt% graphene, and their morphology, wettability, and conductivity were characterized. Mesenchymal stem cells (MSCs) viability, proliferation, adhesion, and orientation on the substrates were evaluated before ES treatment. Then, ES at a frequency of 2 Hz and field strengths of 10, 20, or 50 mV/cm were applied to the cells for 10 min daily, over 3 or 7 consecutive days. Neural differentiation was assessed by morphological observation, immunofluorescence staining, and analysis of neural lineage-related markers. ResultsThe oriented microgroove PDMS substrates supported long-term MSCs growth, guided cells to align along the groove direction, and promoted a morphological transition toward neural-like cells, exhibiting the capacity to induce MSCs differentiation toward the neural lineage. Microgroove structures effectively guided the alignment of MSCs and promoted the development of a more elongated, neural-like morphology, with the most pronounced morphological changes observed on grooves of 10 and 20 μm in width. Meanwhile, the incorporation of graphene endowed the GCPSs with favorable electrical conductivity, though the groove width did not significantly affect the conductive performance. The GCPSs showed good cytocompatibility with MSCs, and cell proliferation on these substrates exhibited a time-dependent pattern, with no significant effect observed from groove width. The inductive effect of ES combined with GCPSs on MSCs differentiation toward specific neural lineages was co-regulated by ES intensity (0, 10, 20, and 50 mV/cm) and microgroove width. ES at 10 mV/cm and 20 mV/cm promotes MSCs differentiation toward neurons on microgrooved substrates of various widths (5, 10, and 20 μm); under the same ES intensities, microgrooves with widths of 5 μm and 10 μm are more favorable for MSCs differentiation toward glial cells/SCs. Notably, low-intensity ES (10 or 20 mV/cm) combined with GCPS 10 significantly enhanced the neural differentiation of MSCs. Neurotrophic factor secretion levels by MSCs cultured on various GCPS substrates after 3 d of ES application was further detected. There were no significant differences in nerve growth factor‑β (NGF‑β) and brain-derived neurotrophic factor (BDNF) concentrations among the GCPS 5, GCPS 10, and GCPS 20 substrates, and these levels were not affected by variations in ES intensity. Under ES at 10 mV/cm, the glial cell line-derived neurotrophic factor (GDNF) concentration in each GCPS substrate was higher than that in the corresponding non-ES group on the same substrate. However, as ES intensity increased, the GDNF concentration decreased to varying degrees. ConclusionThis study demonstrates that the combined strategy of “oriented topographical structure+ES” can effectively promote the neural differentiation of MSCs and holds promise for accelerating the repair process of peripheral nerve injuries.

AIM To study the chemical constituents from Radix Puerariae Lobatae from Anhui province and their in vitro lipid-lowering activity.METHODS D101 macroporous resin,silica gel,MCI,and semi-preparative HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The lipid accumulation model was established by palmitic acid-induced hepatocyte AML12,and the in vitro lipid-lowering activity was evaluated.RESULTS Twenty-one compounds were isolated and identified as carboxymethyl isoferulate(1),methyl(E)feruloylglycolate(2),ariscucurbin-A(3),trans-p-coumaroygly colic acid(4),indole-3-carboxaldehyde(5),β-sitosterol(6),puerol A(7),pueroside C(8),cumoestrol(9),dihydrofurocoumarin(10),decuroside V(11),psoralen dimer(12),bergapten dimer(13),isoliquiritigenin(14),neoliquiritin(15),calycosin(16),daidzein(17),3,4,7-trihydroxyisoflavone(18),ononin(19),genistein(20),genistin(21).Compounds 1-4、7-10、13 and 17-19 significantly inhibited lipid accumulation in AML12 cells,and compound 9 inhibited PPAR-γ expression.CONCLUSION Compounds 1-6,10-13 are isolated from Pueraria genus for the first time.Compounds 1-4,7-10,13 and 17-19 have good lipid-lowering activity.

Objective To establish a chromatography-tandem mass spectrometry method for detecting chlorfenapyr and its metabolite tralopyril in blood,and to investigate the toxicokinetics in rats.Methods Chlorfenapyr(8 mg/kg)was administered orally to rats,and blood samples were collected from rats'canthus vein at 5 min,15 min,30 min,1 h,3 h,6 h,12 h,24 h and 48 h after administration.The blood samples were extracted using 100 μL of 5%formic acid solution and 400 μL of acetonitrile.Chlorfena-pyr was qualitatively and quantitatively detected by triple quadrupole gas chromatography-tandem mass spectrometry(GC-MS/MS)and tralopyril was detected by triple quadrupole liquid chromatography-tandem mass spectrometry(LC-MS/MS).The DAS 3.0 software was used to fit the toxicokinetic equa-tions and calculate the toxicokinetic parameters.Results Chlorfenapyr was detectable from 5 min to 24 h with a peak time of 1 h.Tralopyril was detectable from 15 min to 48 h with a peak time of 3 h.The toxicokinetic process of chlorfenapyr in rat blood conformed to a first-order absorption one-compartment open model,with the toxicokinetic equation described as C=e-0.265t-e-0.175t.Tralopyril con-formed to the first-order absorption three-compartment model,and the toxicokinetic equation was C=47 361.069e-2.209t-35 404.962e-1.486t+11 956.363e-0.512t.In the equations,C stands for the concentration of the target substance in the blood,e is the natural constant(≈2.718 28),and t stands for time.Conclu-sion This study optimized the detection method for chlorfenapyr and its metabolite tralopyril in blood.The toxicokinetic equations and parameters of chlorfenapyr and tralopyril can provide a reference for the estimation of oral intake time of chlorfenapyr.

Objective To examine the changing prevalence and antimicrobial resistance profiles of Burkholderia cepacia in 52 hospitals across China from 2015 to 2021.Methods A total of 9 261 strains of B.cepacia were collected from 52 hospitals between January 1,2015 and December 31,2021.Antimicrobial susceptibility of the strains was tested using Kirby-Bauer method or automated antimicrobial susceptibility testing systems according to a unified protocol.The results were interpreted according to the breakpoints released in the Clinical & Laboratory Standards Institute(CLSI)guidelines(2023 edition).Results A total of 9 261 strains of B.cepacia were isolated from all age groups,especially elderly patients.The proportion was 11.1％(1 032 strains)in children,significantly lower than the proportion in adults.About half(46.5％,4 310/9 261)of the strains were isolated from patients at least 60 years old and 42.3％(3 919/9 261)of the strains were isolated from young adults.Most isolates(71.1％)were isolated from sputum and respiratory secretions,followed by urine(10.7％)and blood samples(8.1％).B.cepacia isolates were highly susceptible to the five antimicrobial agents recommended in the CLSI M100 document(33rd edition,2023).B.cepacia isolates showed relatively higher resistance rates to meropenem and levofloxacin.However,the resistance rates to ceftazidime,trimethoprim-sulfamethoxazole,and minocycline remained below 8.1％.The percentage of B.cepacia strains resistant to levofloxacin was the highest compared to other antibiotics in any of the three age groups(from 12.4％in the patients＜18 years old to 20.6％in the patients aged 60 years or older).Conclusions B.cepacia is one of the clinically important non-fermenting gram-negative bacteria.Accurate and timely reporting of antimicrobial susceptibility test results and ongoing antimicrobial resistance surveillance are helpful for rational prescription of antimicrobial agents and proper prevention and control of nosocomial infections.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.

Objective To characterize the changing species distribution and antibiotic resistance profiles of respiratory isolates in hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Methods Commercial automated antimicrobial susceptibility testing systems and disk diffusion method were used to test the susceptibility of respiratory bacterial isolates to antimicrobial agents following the standardized technical protocol established by the CHINET program.Results A total of 589 746 respiratory isolates were collected from 2015 to 2021.Overall,82.6％of the isolates were Gram-negative bacteria and 17.4％were Gram-positive bacteria.The bacterial isolates from outpatients and inpatients accounted for(6.0±0.9)％and(94.0±0.1)％,respectively.The top microorganisms were Klebsiella spp.,Acinetobacter spp.,Pseudomonas aeruginosa,Staphylococcus aureus,Haemophilus spp.,Stenotrophomonas maltophilia,Escherichia coli,and Streptococcus pneumoniae.Each microorganism was isolated from significantly more males than from females(P＜0.05).The overall prevalence of methicillin-resistant S.aureus(MRSA)was 39.9％.The prevalence of penicillin-resistant S.pneumoniae was 1.4％.The prevalence of extended-spectrum β-lactamase(ESBL)-producing E.coli and K.pneumoniae was 67.8％and 41.3％,respectively.The overall prevalence of carbapenem-resistant E.coli,K.pneumoniae,Enterobacter cloacae,Pseudomonas aeruginosa,and Acinetobacter baumannii was 3.7％,20.8％,9.4％,29.8％,and 73.3％,respectively.The prevalence of β-lactamase was 96.1％in Moraxella catarrhalis and 60.0％in Haemophilus influenzae.The H.influenzae isolates from children(＜18 years)showed significantly higher resistance rates to β-lactam antibiotics than the isolates from adults(P＜0.05).Conclusions Gram-negative bacteria are still predominant in respiratory isolates associated with serious antibiotic resistance.Antimicrobial resistance surveillance should be strengthened in clinical practice to support accurate etiological diagnosis and appropriate antimicrobial therapy based on antimicrobial susceptibility testing results.

AIM To study the chemical constituents from Radix Puerariae Lobatae from Anhui province and their in vitro lipid-lowering activity.METHODS D101 macroporous resin,silica gel,MCI,and semi-preparative HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The lipid accumulation model was established by palmitic acid-induced hepatocyte AML12,and the in vitro lipid-lowering activity was evaluated.RESULTS Twenty-one compounds were isolated and identified as carboxymethyl isoferulate(1),methyl(E)feruloylglycolate(2),ariscucurbin-A(3),trans-p-coumaroygly colic acid(4),indole-3-carboxaldehyde(5),β-sitosterol(6),puerol A(7),pueroside C(8),cumoestrol(9),dihydrofurocoumarin(10),decuroside V(11),psoralen dimer(12),bergapten dimer(13),isoliquiritigenin(14),neoliquiritin(15),calycosin(16),daidzein(17),3,4,7-trihydroxyisoflavone(18),ononin(19),genistein(20),genistin(21).Compounds 1-4、7-10、13 and 17-19 significantly inhibited lipid accumulation in AML12 cells,and compound 9 inhibited PPAR-γ expression.CONCLUSION Compounds 1-6,10-13 are isolated from Pueraria genus for the first time.Compounds 1-4,7-10,13 and 17-19 have good lipid-lowering activity.

Objective:To investigate the nutritional risk status of children in PICU and analyze its influencing factors.Methods:From July 2021 to February 2023,all children aged 1 to 18 years admitted to PICU of Beijing Children's Hospital were investigated by using the pediatric Yorkhill Malnutrition Scoring tool(PYMS)and the clinical data questionnaire.Results:A total of 492 children in PICU were enrolled.The first nutritional risk screening results showed that there were 32 cases of no/low nutritional risk(6.5%),76 cases of medium risk(15.4%),and 384 cases of high risk(78.1%).The incidence of medium/high nutritional risk was as high as 93.5%.The PYMS score of nutritional risk in PICU was(2.61±1.42).The results of multiple linear regression analysis showed that weight,fever time before admission,white blood cells,body mass index,primary diagnosis,father's education,and diet before illness were the main influencing factors of nutritional risk of children in PICU(P<0.05).Conclusion:Children in PICU are in a state of high nutritional risk.It is suggested that children in PICU should carry out nutritional screening in a standardized manner,identify children with high nutritional risk and its influencing factors early.To actively conduct nutritional assessment and nutritional intervention could improve the clinical outcome of children in PICU.

OBJECTIVES: To investigate the synergistic mechanism of the traditional Chinese medicine Rosa laevigata Michx. (RLM) for treatment of pulmonary arterial hypertension (PAH). METHODS: Network pharmacological analysis was carried out to screen the active ingredients of RLM and PAH disease targets and construct the "component-target-disease" interaction network, followed by gene enrichment analysis and molecular docking studies. In the cell experiments, primary cultures of rat pulmonary arterial smooth muscle cells were exposed to hypoxia for 24 h and treated with solvent or 100, 200 and 300 mg/mL RLM, and the changes in cell proliferation were detected using Western blotting for PCNA and immunofluorescence staining. In the animal experiment, male SD rats were randomized into 5 control group, monocrotaline (MCT) solvent group, and MCT with RLM (100, 200 and 300 mg/mL) treatment groups. HE staining and immunofluorescence staining were used to observe histopathological changes in the pulmonary blood vessels of the rats. RESULTS: Seven core active ingredients (including β-sitosterol and kaempferol) in RLM and 39 key disease targets were identified, and molecular docking showed that SRC was a high-affinity target. KEGG enrichment analysis showed that the differential genes were significantly enriched in calcium signaling and PI3K-AKT pathways. In rat pulmonary arterial smooth muscle cells, hypoxic exposure significantly up-regulated cellular expression of PCNA and phosphorylation levels of Src and AKT1, which were obviously lowered by RLM treatment. In RLM-treated rat models, the mean pulmonary artery pressure and right ventricular hypertrophy index (Fulton index) were significantly reduced, the tricuspid annular plane systolic excursion (TAPSE) was improved, and pulmonary vascular wall thickening and fibrosis were obviously ameliorated. CONCLUSIONS RLM inhibits pulmonary arterial smooth muscle cell proliferation in rat models of hypertension possibly by regulating the Src-AKT1 axis, suggesting the potential of RLM as a new natural drug for treatment of pulmonary hypertension.
Animals ; Cell Proliferation/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Rats, Sprague-Dawley ; Pulmonary Artery/cytology* ; Male ; Rats ; Myocytes, Smooth Muscle/cytology* ; Hypertension, Pulmonary/pathology* ; Drugs, Chinese Herbal/pharmacology* ; Signal Transduction/drug effects* ; Muscle, Smooth, Vascular/cytology* ; src-Family Kinases/metabolism* ; Cells, Cultured

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.