OBJECTIVE To investigate the mechanism of Xihuang pill （XHP） against diffuse large B-cell lymphoma （DLBCL）. METHODS The active ingredients of XHP and potential therapeutic targets for DLBCL were identified using TCMSP， GeneCards and DisGeNET databases. Protein-protein interaction networks were constructed using the String database and Cytoscape software to screen core components and core targets. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were then performed. The clinical relevance of core targets was analyzed using the GEPIA and PanCanSurvPlot databases. Molecular docking and molecular dynamics （MD） simulation were conducted to verify the interactions between core components and core targets， and the binding free energy was calculated using the molecular mechanics Poisson-Boltzmann surface area （MM-PBSA） method. The effects of XHP on DLBCL and the related molecular mechanisms were validated using CCK-8 assay， flow cytometry and Western blot. RESULTS Network pharmacology analysis identified 108 active ingredients of XHP and 410 potential therapeutic targets for DLBCL. Six core components （e.g.， 17 beta-estradiol， quercetin） and ten core targets [e.g.， tumor protein 53 （TP53）， proto-oncogene tyrosine-protein kinase Src （SRC）] were obtained. Enrichment analysis indicated that the anti-DLBCL effects of XHP were primarily associated with the apoptotic signaling pathway， the phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway and so on. Clinical correlation analysis revealed that TP53 and SRC expression were significantly up-regulated in DLBCL tissues and associated with poor patient prognosis （P＜0.05）. Molecular docking， MD simulations and MM-PBSA calculations confirmed that the SRC-quercetin complex had a mail：stronger and more stable binding affinity. In vitro experiments demonstrated that XHP concentration-dependently inhibited the proliferation of DLBCL cells； compared with control group， XHP medium- and high-dose groups could significantly induce the apoptosis of SU-DHL2 and SU-DHL4 cells， and significantly down- regulated the expressions of SRC protein， phosphorylated （p）-PI3K/PI3K and p-Akt/Akt in SU-DHL4 cells （P＜0.05）. CONCLUSIONS XHP may inhibit the proliferation and induce the apoptosis of DLBCL cells by regulating the SRC/PI3K/Akt signaling pathway.

Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10 /mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.

ObjectiveTo investigate the effects of chronic hypoxic exposure on lipid metabolism in metabolic dysfunction-associated steatohepatitis mice via transcriptomic analysis. MethodsMale C57BL/6J mice were randomly divided into 3 groups： Control group （normal diet）， metabolic dysfunction-associated steatohepatitis group （MASH， high-fat diet）， hypoxic metabolic dysfunction-associated steatohepatitis group （CH-MASH， high-fat diet）. After 12 weeks， CH-MASH was given hypoxia intervention （altitude=3 800 m） for 4 weeks. Differentially expressed genes in the livers of mice from the three groups were identified using transcriptome sequencing， followed by GO and KEGG analyses. A protein-protein interaction （PPI） network was constructed to screen hub genes. Serum triglyceride （TG）， total cholesterol （TC）， and liver function parameters were measured， and hepatic histomorphological changes were observed by HE staining. ResultsSerum TG and TC significantly increased in MASH group and CH-MASH group compared with Control group. Compared to MASH group， serum TG and TC obviously increased in CH-MASH. HE staining suggested that the lipid level of CH-MASH was reduced compared with MASH. Liver transcriptomes differed markedly between CH-MASH and MASH mice. A total of 1 005 differentially expressed genes were screened. A total of 1 151 significantly enriched GO terms were obtained including extracellular regions， intrinsic components of membranes， lipid metabolism processes， small molecule metabolism processes， and small molecule synthesis processes KEGG enrichment analysis showed that DEGs were enriched in 41 pathways， including metabolic pathway， retinol metabolic pathway， PPAR signaling pathway， sterol synthesis pathway etc. In the screening protein interaction network， the closely connected genes related to lipid metabolism under hypoxia were Acaca， Hmgcs1， Acat2， Cyp51和Msmo1. The results of RT-qPCR were consistent. ConclusionChronic hypoxia significantly alters liver lipid metabolism transcription levels in MASH mice. The key genes and pathways screened in this study provide clues for further research on the mechanism of hypoxia exposure affecting liver metabolic changes in MASH mice.

Objective To investigate the correlation of monocyte count(MONO)to high density lipopro-tein-cholesterol(HDL-C)ratio(MHR)and systemic immune-inflammation index(SII)with diabetic retinop-athy(DR).Methods Patients with type 2 diabetes mellitus(T2DM)admitted to the hospital from June 2020 to May 2023 were selected as the research objects.According to the presence or absence of DR,the patients were divided into non-retinopathy group(NDR group)and DR Group.The differences in basic information,blood routine,and biochemical indexes between the two groups were analyzed,and the MHR and SII were cal-culated.Multivariate Logistic regression was used to analyze the risk factors for DR.Spearman correlation a-nalysis was used to analyze the correlation between risk factors and DR.The receiver operating characteristic(ROC)curve was used to evaluate the value of MHR and SII in predicting DR in T2DM patients.Results A total of 291 T2DM patients were enrolled,including 135 patients in the NDR group and 156 patients in the DR group.Compared with the NDR group,duration of diabetes was significantly prolonged(P<0.05),glycosy-lated hemoglobin(HbA1c),creatinine,fasting plasma glucose(FPG),total cholesterol(TC),platelet count(PLT),MHR and SII were increased(P<0.05),and high density lipoprotein-cholesterol(HDL-C)was de-creased(P<0.05)in the DR Group.Spearman correlation analysis showed that DR was positively correlated with duration of diabetes,FPG,HbA1c,PLT,MHR and SII(P<0.05),and negatively correlated with HDL-C(P<0.05).Multivariate Logistic regression analysis showed that gender(OR=0.151,95%CI 0.052-0.432,P<0.001),history of heavy drinking(OR=7.199,95%CI 2.845-18.216,P<0.001),duration of di-abetes(OR=1.570,95%CI 1.354-1.821,P<0.001),HbA1c(OR=1.218,95%CI 1.013-1.464,P=0.036),MHR(OR=1.054,95%CI 1.028-1.080,P<0.001)and SII(OR=1.002,95%CI 1.001-1.003,P=0.002)were independent influencing factors for DR patients.ROC curve analysis showed that the area un-der the curve(AUC)of MHR and SII in predicting the development of T2DM to DR was 0.696 and 0.567,re-spectively.The AUC of MHR and SII combined in predicting DR was 0.702.Conclusion MHR and SII are closely related to the incidence of DR,and both have certain predictive value for DR,and the predictive value of the combined of MHR and SII is higher.

Ursodeoxycholic acid(UDCA)is a naturally occurring,low-toxicity,and hydrophilic bile acid(BA)in the human body that is converted by intestinal flora using primary BA.Solute carrier family 7 member 11(SLC7A11)functions to uptake extracellular cystine in exchange for glutamate,and is highly expressed in a variety of human cancers.Retroperitoneal liposarcoma(RLPS)refers to liposarcoma originating from the retroperitoneal area.Lipidomics analysis revealed that UDCA was one of the most significantly down-regulated metabolites in sera of RIPS patients compared with healthy subjects.The augmentation of UDCA concentration(≥25 μg/mL)demonstrated a suppressive effect on the proliferation of liposarcoma cells.[15N2]-cystine and[13Cs]-glutamine isotope tracing revealed that UDCA impairs cystine uptake and glutathione(GSH)synthesis.Mechanistically,UDCA binds to the cystine transporter SLC7A11 to inhibit cystine uptake and impair GSH de novo synthesis,leading to reactive oxygen species(ROS)accumulation and mitochondrial oxidative damage.Furthermore,UDCA can promote the anti-cancer effects of ferroptosis inducers(Erastin,RSL3),the murine double minute 2(MDM2)inhibitors(Nutlin 3a,RG7112),cyclin dependent kinase 4(CDK4)inhibitor(Abemaciclib),and glutaminase inhibitor(CB839).Together,UDCA functions as a cystine exchange factor that binds to SLC7A11 for antitumor activity,and SLC7A11 is not only a new transporter for BA but also a clinically applicable target for UDCA.More importantly,in combination with other antitumor chemotherapy or physiotherapy treatments,UDCA may provide effective and promising treatment strategies for RLPS or other types of tumors in a ROS-dependent manner.

Objective Advances in synthetic DNA technology have made it much easier to fake human DNA samples.There are literature reports that fake human DNA can be synthesized by different methods and implanted in the field to confuse the investigation or mislead the trial.Therefore,distinguishing authentic human DNA from synthetic DNA and performing individual identification has become a critical scientific challenge.Methods We define a novel composite genetic marker(methylation-microhaplotype)by combining CpG sites stably hypermethylated or hypomethylated in natural human DNA and nearby immediately adjacent microhaplotype sites.A total of 19 locis were obtained according to the screening criteria,and a composite detection system for methylation-microhaplotypes was established using MPS technology.Random volunteer DNA samples were extracted and synthetic DNA samples were prepared based on whole genome amplification techniques.Population DNA samples were analyzed to evaluate forensic parameters and methylation variability of the methylation-microhaplotype markers.Comparative analyses of human and synthetic DNA were conducted to assess the markers'ability to discriminate between the two and to detect/type both components in mixed mixed samples.Results The composite detection system composed of 19 locis demonstrated high individual identification ability,achieving a cumulative individual identification probability of 0.999 999 999 996 86.12 hypermethylated locis and 7 hypomethylated locis had relatively stable methylation levels in 57 human DNA samples.According to the allele methylation rate(Ram)value,the system can effectively identify natural and synthetic DNA samples.Meanwhile,for mixed DNA samples,the presence of human and synthetic DNA samples can be found and genotyped.Conclusion Methylation-microhaplotype genetic markers,which can discover human DNA and synthetic DNA and can detect the presence and genotyping of them from mixed samples,is a potential useful tool for forensic DNA analysis.

Enteric human adenovirus(HAdV),a common cause of acute viral gastroenteritis in children,frequently triggers spo-radic infections,nosocomial transmissions,and outbreaks in kindergarten settings.This study was aimed at investigating the molecular characteristics and genetic evolution of enteric HAdV among patients with acute gastroenteritis younger than 5 years in China,to pro-vide foundational data for disease prevention and control.A total of 8 074 stool samples were collected from hospitalized or outpatient children younger than 5 years with acute gastroenteritis in China during 2023.HAdV screening was conducted with real-time fluores-cence PCR.Positive samples were sequenced,then subjected to bioinformatics analysis including genotyping,homology assessment,and phylogenetic analysis with GenBank,BioAider,and MEGA11.0.A total of 370 samples(4.58%)tested positive for HAdV.Two enteric HAdV genotypes were identified:HAdV-F41(which predominated,at 98.09%)and HAdV-F40(1.90%).HAdV-F41 was the dominant genotype among patients with acute gastroenteritis younger than 5 years in China.Bioinformatics analysis indicated that the predominant HAdV lineages in China were lineage 1 and 2,whereas European lineage 3 showed no influence.Systematic and long-term surveillance of HAdV should help elucidate its diversity and evolutionary patterns in China,thereby providing scientific evi-dence for developing more effective prevention strategies.

This study investigated the full-genome molecular characteristics of a rotavirus vaccine-derived strain,G1P[8]geno-type A group rotavirus RVA/Human-wt/CHN/HN1140/2021/G1P[8](referred to as HN1140).The gene fragments of the HN1140 strain were amplified with reverse transcription-polymerase chain reaction(RT-PCR)combined with whole-genome primers to obtain the full genome sequence.Genotyping was performed with the online genotyping tool RotaC 2.0,and similarity and genetic evolution analyses for each gene segment were conducted in DNAstar5.1 and MEGA11.0 software.The genotype of the HN1140 strain was deter-mined to be G1-P[8]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis demonstrated that all 11 genomic segments clus-tered closely with the RotaTeq vaccine strains,sharing 99.7%-100%nucleotide sequence similarity.Notably,VP1,VP2,VP6,and NSP2-NSP5 segments showed 100%nucleotide identity with RotaTeq strains.Comparative genomic analysis identified 13 nucleotide and 8 amino acid substitutions between HN1140 and RotaTeq strains,localized within the VP7,VP4,VP1,VP2,VP3,and NSP1 segments.The HN1140 strain exhibited the genotype G1-P[8]-A3-T6-H3,which was consistent with the typical profile of a vaccine-derived reassortant.This strain demonstrated high genetic similarity to RotaTeq vaccine strains,with nucleotide sequence identity ranging from 99.7%to 100%.These findings suggested that HN1140 evolved from RotaTeq vaccine strains through genetic reassortment.

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.

OBJECTIVES: This study aims to use 3D prin-ting technology based on the principle of stereo lithography apparatus (SLA) to shape dental lithium disilicate ceramics and study the effects of different slurry proportions on the microstructure and properties of heat-treated samples. METHODS: The experimental group comprised lithium disilicate ceramics manufactured through SLA 3D printing, and the control group comprised lithium disilicate ceramics (IPS e.max CAD) fabricated through commercial milling. An array of different particle sizes of lithium disilicate ceramic powder materials (nano and micron) was selected for mixing with photocurable acrylate resin. The proportion of experimental raw materials was adjusted to prepare five groups of ceramic slurries for 3D printing (Groups S1-S5) on the basis of rheological properties, stability, and other factors. Printing, debonding, and sintering were conducted on the experimental group with the optimal ratio, followed by measurements of microstructure, crystallographic information, shrinkage, and mechanical properties. RESULTS: Five groups of lithium disilicate ceramic slurries were prepared, of which two groups with high solid content (75%) (Groups S2 and S3) were selected for 3D printing. X-ray diffraction and scanning electron microscopy results showed that lithium disilicate was the main crystalline phase in Groups S2 and S3, and its microstructure was slender, uniform, and compact. The average grain sizes of Groups S2 and S3 were (559.79±84.58) nm and (388.26±61.49) nm, respectively (P<0.05). Energy spectroscopy revealed that the samples in the two groups contained a high proportion of Si and O elements. After heat treatment, the shrinkage rate of the two groups of ceramic samples was 18.00%-20.71%. Test results revealed no statistical difference in all mechanical properties between Groups S2 and S3 (P>0.05). The flexural strengths of Groups S2 and S3 were (231.79±21.71) MPa and (214.86±46.64) MPa, respectively, which were lower than that of the IPS e.max CAD group (P<0.05). The elasticity modulus of Groups S2 and S3 were (87.40±12.99) GPa and (92.87±19.76) GPa, respectively, which did not significantly differ from that of the IPS e.max CAD group (P>0.05). The Vickers hardness values of Groups S2 and S3 were (6.53±0.19) GPa and (6.25±0.12) GPa, respectively, which were higher than that of the IPS e.max CAD group (P<0.05). The fracture toughness values of Groups S2 and S3 were (1.57±0.28) MPa·m^0.5 and (1.38±0.17) MPa·m^0.5, respectively, which did not significantly differ from that of the IPS e.max CAD group (P>0.05). CONCLUSIONS The combination of lithium disilicate ceramic powders with different particle sizes can yield a slurry with high solid content (75%) and suitable viscosity and stability. The dental lithium disilicate ceramic material is successfully prepared by using 3D printing technology. The 3D-printed samples show a small shrinkage rate after heat treatment. Their microstructure conforms to the crystal phase of lithium disilicate ceramics, and their mechanical properties are close to those of milled lithium disilicate ceramics.
Printing, Three-Dimensional ; Dental Porcelain/chemistry* ; Ceramics/chemistry* ; Materials Testing ; Particle Size