AIM To investigate the clinical effects of Cinobufosin Injection combined with RALOX-HAIC regimen on patients with hepatocellular carcinoma.METHODS Ninety-two patients were randomly assigned into control group(46 cases)for intervention of RALOX-HAIC regimen,and observation group(46 cases)for intervention of both Cinobufosin Injection and RALOX-HAIC regimen.The changes in short-term effects,survival situation,inflammatory indices(LCN2,NLRP3 inflammasome,NLR,PLR),immune indices(NK cells,CD8+T cells,IL-17,Th17/Treg)and incidence of toxic and side effects were detected.RESULTS Based on mRECIST,the observation group demonstrated higher disease control rate and objective remission rate than the control group(P＜0.05),along with lower disease progression(P＜0.05).After the treatment,the two groups displayed decreased inflammatory indices,IL-17,Th17/Treg(P＜0.05),and increased NK cells,CD8+T cells(P＜0.05),especially for the observation group(P＜0.05).The observation group exhibited lower incidence of abdominal pain,nausea,vomiting,diarrhea,leukopenia and thrombocytopenia than the control group(P＜0.05),and no significant differences in overall survival and incidence of other toxic and side effects were found between the two groups(P＞0.05).CONCLUSION For the patients with hepatocellular carcinoma,Cinobufosin Injection combined with RALOX-HAIC regimen can safely and effectively enhance body immune functions,and reduce in vivo immune indices.

Surgical intervention for malignant bone tumors frequently results in bone defects located at the metaphysis of the long bones in the lower extremities.The morphological heterogeneity of the metaphysis poses significant challenges for conventional treatment methods to adequately conform to the defect area.The utilization of three-dimensional(3D)-printed titanium bone repair scaffolds has emerged as an effective reconstructive approach for metaphyseal bone defects,as these scaffolds offer precise shape conformity and provide adequate mechanical support.However,the current commonly used scaffolds do not adequately replicate the biomechanical environment of bone defects,resulting in suboptimal bone ingrowth within the scaffolds and subsequent prosthesis loosening and failure post-operation.Bone is a highly force-responsive organ,and its fate is regulated by biomechanical signals.Consequently,designing scaffolds with consideration of biomechanical principles to ensure mechanical compatibility between the scaffolds and the bone defect sites is a critical factor influencing the success of bone defects reconstruction.This review primarily introduces the biomechanical factors influencing bone defect repair and the advancements in designing 3D-printed titanium bone repair scaffolds biomechanically matched with bones,offering theoretical guidance for scaffold design and preparation.

Objective:To investigate the effects of exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) during osteogenic differentiation on the polarization of Raw264.7 macrophages and to elucidate the underlying mechanisms.Methods:PBS, uninduced BMSCs conditioned medium (CM), and osteogenic induction BMSCs CM for 7 d, 14 d, and 21 d were respectively added to Raw264.7 macrophages. After 48 h of treatment, Western blotting was used to detect and compare the expression of M1 markers of macrophages [inducible nitric oxide synthase (iNOS) and CD86] and M2 markers of macrophages [arginase-1 (ARG-1) and CD163] in each group. In addition, Raw264.7 macrophages were divided into three groups: the PBS group (only PBS added), the BMSCs-exo group (exosomes derived from uninduced BMSCs were added), and 7D-BMSCs-exo (exosomes derived from BMSCs were added after 7 days of osteogenic induction). The absorbance values of Raw264.7 cells in each group at 24 h, 48 h and 72 h were detected by an enzyme-linked immunosorbent assay (ELISA) reader. Western blotting was performed to assess changes of M1 or M2 marker proteins in Raw264.7 macrophages treated by exosome. The supernatants of the three groups of Raw264.7 macrophages were then co-cultured with BMSCs. Alizarin Red staining was used to quantify the formation of mineralized nodules, and alkaline phosphatase (ALP) staining was used to evaluate the osteogenic activity, and the expression levels of osteogenesis-related proteins Runx2, ALP, osteopontin (OPN), and osteocalcin (OCN) were detected. The migration ability of endothelial progenitor cells (EPCs) was detected by scratch assay for migration distance, and the angiogenesis ability was detected by in vitro tube formation assay for the number of vascular rings formed. The expressions of vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF) were detected by Western blot. The activation of the MAPK signaling pathway was evaluated by measuring phosphorylated and total P38 and ERK1/2 levels.Results:Western blot analysis showed that CM from BMSCs after 7 days of osteogenic induction (7D-BMSCs-CM) induced the strongest M2 polarization in macrophages. Compared with the PBS group, 7D-BMSCs-CM induced the most significant polarization of Raw264.7 macrophages to the M2 type, and increased ARG-1 and CD163 expression to 1.36±0.09 and 1.69±0.09, respectively ( P<0.05), while decreasing iNOS and CD86 to 0.21±0.03 and 0.29±0.03 ( P<0.05). The absorbance values of macrophages in the 7D-BMSCs-exo group were significantly higher than those in the PBS group at 24 h, 48 h, and 72 h ( P<0.05). Compared with BMSCs-exo group, 7D-BMSCs-exo upregulated the expressions of CD163 and ARG-1 while inhibited the expressions of iNOS and CD86 ( P<0.05). Alizarin red staining showed enhanced mineral deposition and a higher degree of mineralization in the 7D-BMSCs-exo group, the staining intensity of ALP also increased simultaneously, the Western-blot results showed that the protein expressions of Runx2, ALP, OPN and OCN were respectively higher than those in the PBS group ( P<0.05). The results of the scratch assay showed that the relative migration distance of EPCs cells in the 7D-BMSCS-exo group at 24 h reached 2.30±0.05 μm, which was higher than that in the PBS group 1.10±0.02 μm ( P<0.05). The Tube formation experiment showed that the number of vascular rings in the EPCs group was higher than that in the PBS group (30.3±2.5 and 15.0±1.0, P<0.05), and the protein expressions of VEGF-A and PDGF were upregulated. Furthermore, the levels of phosphorylated P38 and ERK1/2 were significantly reduced in the 7D-BMSCs-exo group 0.40±0.06 and 0.25±0.06 compared with the PBS group ( P<0.05). Conclusions:Exosomes secreted by BMSCs during osteogenic differentiation promote the M2 polarization of Raw264.7 macrophages, with the most pronounced effect observed at the 7th day. M2-polarized macrophages, in turn, enhance the osteogenic differentiation of BMSCs and the angiogenic capacity of EPCs. These processes are closely associated with the suppression of the MAPK signaling pathway.

Objective:To understand the etiological surveillance and drug resistance characteristics of Legionella pneumophila (LP) from the aqueous environment of public places in Shanghai, from 2011 to 2020, and provide evidence for surveillance of the disease. Methods:Environmental water samples were systematically collected from public venues in urban and suburban districts of Shanghai for LP surveillance. All the identified LP isolates underwent a series of testings including serotyping, pulsed field gel electrophoresis (PFGE), sequence-based typing, and antimicrobial susceptibility testing. χ2 test or Cochran-Armitage trend tests were used for statistical analysis and for temporal resistance patterns. Results:Among 6 263 water samples, the LP-positive rate was 20.93% (1 311/6 263). The positivity rate decreased from 24.98% (287/1 149) in 2011-2012 to 20.02% (1 024/5 114) in 2013-2020 ( χ2=13.92, P<0.001), with the highest monthly positivity observed from June to August (23.79%, 745/3 132). A total of 1 365 LP strains were isolated, of which 912 were further characterized, including 10 serotypes, 149 PFGE patterns, and 33 sequence types (ST). The predominant serotype was Lp1 (86.84%, 792/912), and the dominant ST was ST752 (29.50%, 269/912). ST clustering revealed two major clonal groups CG1 and CG2, accounting for 91.12% (831/912) of the isolates. The 190 LPs involved in the drug sensitivity test showed three resistance profiles: azithromycin resistance (31.05%, 59/190), ciprofloxacin resistance (0.53%, 1/190) and azithromycin+ciprofloxacin resistance (0.53%, 1/190). Azithromycin-resistant strains were predominantly ST1 (64.41%, 38/59). The antimicrobial resistance rate showed a significant decline, from 48.65% (18/37) in 2011-2012 to 28.10% (43/153) in 2013-2020 ( χ2=9.38, P=0.002). Conclusions:Compared to from 2011 to 2012, both the positivity rate and antimicrobial resistance prevalence of LP in public aqueous environments of Shanghai exhibited an overall decline from 2013 to 2020. The predominant types of LP were serotype Lp1 and sequence type ST752, with notable high-level resistance to azithromycin. Measures as enhancing the enforcement of water safety regulations and prioritizing surveillance of azithromycin resistance in LP were recommended to mitigate public health risks.

[Objective]To investigate the demyelination induced by Angiostrongylus cantonensis(AC)infection in the brain of Balb/c mice and analyze the untargeted metabolomic changes in the corpus callosum,aiming to elucidate the underlying mechanisms.[Methods]Balb/c mice were randomly assigned to a control group(n=6)and an infection group(n=6).The infection group was orally administered 30 third-stage larvae of AC,while the control group received an equal volume of saline.Body weight,visual function,and behavioral scores were measured on post-infection 3,6,9,12,15,18,and 21 days to assess neurological alterations.After 21 days,brain tissues were harvested for immunofluorescence staining,hematoxylin-eosin(HE)staining,and transmission electron microscopy to examine morphological changes in brain myelin and retina.Metabolomics analysis was performed,and differential metabolites were identified using volcano plots and heatmaps.The distribution of fold changes and bar charts were used to profile the key metabolites.These differential metabolites were then subjected to Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and regulatory network analysis.[Results]On the 9th day after AC infection,Balb/c mice showed a decline in neurological behavioral scores(P<0.05).By day 15,visual scores decreased(P<0.05),and by day 21,significant weight loss(P＜0.001)and mortality were observed.Concurrently,transmission electron microscopy and immunofluorescence staining revealed significant myelin damage in the corpus callosum and a marked reduction in oligodendrocytes(P<0.001).HE staining showed severe retinal ganglion cell damage.Metabolomic analysis revealed that glycerophospholipids were the most abundant differential metabolites,with steroids and sphingolipids being relatively less abundant.Cholesteryl ester CE(20:2)was significantly upregulated(P<0.001),while phosphatidylmethanol(18:0_18:1)was significantly downregulated(P<0.01).KEGG enrichment and regulatory network analyses demonstrated that the differential metabolites were mainly enriched in metabolic pathways like steroid biosynthesis,bile secretion,and cholesterol metabolism,and were involved in key metabolic pathways such as sphingolipid metabolism,neural signal regulation,and glycerophospholipid metabolism.[Conclusions]AC infection affects the metabolic state of mice via multiple pathways,modifying the levels of metabolites crucial for myelination and myelin stability.Demyelination may be closely linked to the disruption of these key metabolic pathways,particularly the dysregulation of cholesterol and sphingolipid metabolism,potentially playing a central role in demyelination onset.Furthermore,alterations in phospholipid metabolism and abnormal nerve signaling regulation may exacerbate myelin damage.

To explore the variation laws of volatile oil during the extraction process of Artemisiae Argyi Folium and its impact on the quality of the medicinal solution, as well as to achieve precise control of the extraction process, this study employed headspace solid phase microextraction gas chromatography-mass spectrometry(HS-SPME-GC-MS) in combination with multiple light scattering techniques to conduct a comprehensive analysis, identification, and characterization of the changes in volatile components and the physical properties of the medicinal solution during the extraction process. A total of 82 volatile compounds were identified using the HS-SPME-GC-MS technique, including 21 alcohols, 15 alkenes, 14 ketones, 9 acids, 6 aldehydes, 5 phenols, 3 esters, and 9 other types of compounds. At different extraction time points(15, 30, 45, and 60 min), 71, 72, 64, and 44 compounds were identified in the medicinal solution, respectively. It was observed that the content of volatile components gradually decreased with the extension of extraction time. Through multivariate statistical analysis, four compounds with significant differences during different extraction time intervals were identified, namely 1,8-cineole, terpinen-4-ol, 3-octanone, and camphor. RESULTS:: from multiple light scattering techniques indicated that at 15 minutes of extraction, the transmittance of the medicinal solution was the lowest(25%), the particle size was the largest(0.325-0.350 nm), and the stability index(turbiscan stability index, TSI) was the highest(0-2.5). With the extension of extraction time, the light transmittance of the medicinal solution improved, stability was enhanced, and the particle size decreased. These laws of physicochemical property changes provide important basis for the control of Artemisiae Argyi Folium extraction process. In addition, the changes in the bioactivity of Artemisiae Argyi Folium extracts during the extraction process were investigated through mouse writhing tests and antimicrobial assays. The results indicated that the analgesic and antimicrobial effects of the medicinal solution were strongest at the 15-minute extracting point. In summary, the findings of this study demonstrate that the content of volatile oil in Artemisiae Argyi Folium extracts gradually decreases with the extension of extraction time, and the variation in volatile oil content directly influences the physicochemical properties and pharmacological efficacy of the medicinal solution. This discovery provides important scientific reference for the optimization of Artemisiae Argyi Folium extraction processes and the development and application of process analytical technologies.
Oils, Volatile/pharmacology* ; Artemisia/chemistry* ; Gas Chromatography-Mass Spectrometry ; Drugs, Chinese Herbal/pharmacology* ; Chlorogenic Acid/pharmacology* ; Solid Phase Microextraction ; Quality Control

Microbial detection and control of traditional Chinese medicine(TCM) decoction pieces are crucial for the quality control of TCM preparations. It is also a key area of research in the measurement technology and equipment development for TCM manufacturing. Guided by TCM manufacturing measurement methodologies, this study presented a design of a novel portable microbial detection device, using Atractylodis Macrocephalae Rhizoma decoction pieces as a demonstration. Immunomagnetic separation technology was employed for specific isolation and labeling of target microorganisms. Enzymatic signal amplification was utilized to convert weak biological signals into colorimetric signals, constructing an optical biosensor. A self-developed smartphone APP was further applied to analyze the colorimetric signals and quantify target concentrations. A portable and automated detection system based on Arduino microcontroller was developed to automatically perform target microbial separation/extraction, as well as mimetic enzyme labeling and catalytic reactions. The developed equipment specifically focuses on the rapid and quantitative microbial analysis of TCM active pharmaceutical ingredients, intermediates in TCM manufacturing, and final TCM products. Experimental results demonstrate that the equipment could detect Salmonella in samples within 2 h, with a detection limit as low as 5.1 × 10~3 CFU·mL~(-1). The equipment enables the rapid detection of microorganisms in TCM decoction pieces, providing a potential technical solution for on-site rapid screening of microbial contamination indicators in TCM. It has broad application prospects in measurement technology for TCM manufacturing and offers strong technical support for the modernization, industrialization, and intelligent development of TCM.
Drugs, Chinese Herbal/analysis* ; Atractylodes/microbiology* ; Rhizome/microbiology* ; Biosensing Techniques/methods* ; Medicine, Chinese Traditional ; Colorimetry/instrumentation* ; Quality Control

OBJECTIVE To standardize the strategies for prevention and control of Chikungunya(CHIK)in healthcare in-stitutions so as to reduce the risk of transmission in the institutions.METHODS A working group comprising the ex-perts in hospital infection control,infectious diseases,and microbiology systematically reviewed domestic and international evidence and current guidelines,integrated China's vector ecology and healthcare realities,conducted two rounds of Delphi to achieve expert consensus,and graded the evidence and recommendation strength using the Oxford Centre for Evidence Based Medicine system.RESULTS The consensus issues 18 actionable recommendations on triage,patient mosquito-proof isolation,integrated vector control,protection of susceptible populations,environmental cleaning and disinfection,specimen management,medical textile handling,and outbreak emergency response,with each statement assigned an evi-dence level and recommendation strength.CONCLUSION This consensus is for the first time in China to provide evidence-graded strategies for control of CHIK in healthcare institutions,offering work flow-oriented,implementable guidance for clinicians,laboratorians,and infection-control personnel under different risk scenarios and enhancing the comprehensive coping capacity of the healthcare institutions.

OBJECTIVE: This study reports the first imported case of Lassa fever (LF) in China. Laboratory detection and molecular epidemiological analysis of the Lassa virus (LASV) from this case offer valuable insights for the prevention and control of LF. METHODS: Samples of cerebrospinal fluid (CSF), blood, urine, saliva, and environmental materials were collected from the patient and their close contacts for LASV nucleotide detection. Whole-genome sequencing was performed on positive samples to analyze the genetic characteristics of the virus. RESULTS: LASV was detected in the patient's CSF, blood, and urine, while all samples from close contacts and the environment tested negative. The virus belongs to the lineage IV strain and shares the highest homology with strains from Sierra Leone. The variability in the glycoprotein complex (GPC) among different strains ranged from 3.9% to 15.1%, higher than previously reported for the seven known lineages. Amino acid mutation analysis revealed multiple mutations within the GPC immunogenic epitopes, increasing strain diversity and potentially impacting immune response. CONCLUSION The case was confirmed through nucleotide detection, with no evidence of secondary transmission or viral spread. The LASV strain identified belongs to lineage IV, with broader GPC variability than previously reported. Mutations in the immune-related sites of GPC may affect immune responses, necessitating heightened vigilance regarding the virus.
Humans ; China/epidemiology* ; Genome, Viral ; Lassa Fever/virology* ; Lassa virus/classification* ; Molecular Epidemiology ; Phylogeny

OBJECTIVE: The objective of our study was to evaluate the vaccine effectiveness (VE) of the pentavalent rotavirus vaccine (RV5) among < 5-year-old children in three provinces of China during 2020-2024 via a propensity score-matched test-negative case-control study. METHODS: Electronic health records and immunization information systems were used to obtain data on acute gastroenteritis (AGE) cases tested for rotavirus (RV) infection. RV-positive cases were propensity score matched with RV-negative controls for age, visit month, and province. RESULTS: The study included 27,472 children with AGE aged 8 weeks to 4 years at the time of AGE diagnosis; 7.98% (2,192) were RV-positive. The VE (95% confidence interval, CI) of 1-2 and 3 doses of RV5 against any medically attended RV infection (inpatient or outpatient) was 57.6% (39.8%, 70.2%) and 67.2% (60.3%, 72.9%), respectively. Among children who received the 3rd dose before turning 5 months of age, 3-dose VE decreased from 70.4% (53.9%, 81.1%) (< 5 months since the 3rd dose) to 63.0% (49.1%, 73.0%) (≥ 1 year since the 3rd dose). The three-dose VE rate was 69.4% (41.3%, 84.0%) for RVGE hospitalization and 57.5% (38.9%, 70.5%) for outpatient-only medically attended RVGE. CONCLUSION Three-dose RV5 VE against rotavirus gastroenteritis (RVGE) in children aged < 5 years was higher than 1-2-dose VE. Three-dose VE decreased with time since the 3rd dose in children who received the 3rd dose before turning five months of age, but remained above 60% for at least one year. VE was higher for RVGE hospitalizations than for medically attended outpatient visits.
Humans ; Rotavirus Vaccines/immunology* ; China/epidemiology* ; Case-Control Studies ; Child, Preschool ; Infant ; Rotavirus Infections/epidemiology* ; Male ; Propensity Score ; Female ; Vaccine Efficacy ; Gastroenteritis/virology* ; Vaccines, Attenuated ; Rotavirus