Primary liver cancer is a malignant tumor with continuously rising incidence and mortality rates worldwide， imposing a heavy burden on patients and society， and hepatocellular carcinoma （HCC） is a common type of primary liver cancer. As one of the important treatment methods for HCC， radiotherapy can effectively control the local growth of tumors and alleviate symptoms in patients. However， radiotherapy resistance seriously affects the treatment effect and has become a major challenge in clinical treatment. Current research shows a complex mechanism of radiotherapy resistance in HCC， involving multiple factors such as abnormal activation of intracellular signaling pathways， changes in tumor microenvironment， and regulation of gene expression. Therefore， a series of strategies have been proposed to address radiotherapy resistance in clinical practice， including regulating cell signaling pathways， improving tumor microenvironment， and combining different treatment modalities， and such strategies have shown promising application prospects. This article reviews the research advances in the mechanism of radiotherapy resistance and related coping strategies， in order to provide new perspectives for future research on radiotherapy for HCC.

Objective:To explore the protective effect of sesquiterpene lactones of Eupatorium lindleyanum DC.(SLEL)on lipopolysac-charide(LPS)-induced acute lung injury(ALI)in rats using metabolomics.Methods:Forty-eight male SD rats were randomly divided into a blank control group(CG),a model control group(MG),low-,medium-,and high-dose SLEL groups(50,100,and 200 mg/kg),and a positive control group(dexamethasone acetate tablets,5 mg/kg).CG and MG groups were given phosphate-buffered saline.All groups received intragastric administration at a dose volume of 10 mL/kg once a day for 7 consecutive days.One hour after the last ad-ministration,LPS(5 mg/kg)was instilled into the trachea of all groups except the CG group to establish the ALI rat model.Twenty-four hours after model establishment,blood was collected from the abdominal aorta and bronchoalveolar lavage fluid(BALF)was col-lected from the left lung.The total number of inflammatory cells,neutrophils,lymphocytes,and eosinophils in BALF was counted by Wright-Giemsa staining.The levels of interleukin-18(IL-18)and interferon-γ(IFN-γ)in serum and BALF were measured by enzyme-linked immunosorbent assay.The pathological changes of lung tissue were observed using hematoxylin-eosin staining.The ex-pression levels of tight junction protein-1(ZO-1)and occludin in lung tissue were determined by Western blot.The mRNA expression levels of IL-18,IFN-γ,ZO-1,and occludin in the lung were measured by real-time fluorescence quantitative PCR.Non-targeted me-tabolomics analysis of serum and lung tissue was performed using ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry.Results:Compared with the MG group,all SLEL groups had significantly reduced wet/dry weight ratio of lung tissue,lung coefficient,and total number of inflammatory cells,neutrophils,lymphocytes,and eosinophils in BALF.SLEL signifi-cantly decreased the levels of IL-18 and IFN-γ in serum and BALF and the mRNA expression of IL-18 and IFN-γ in lung tissue,and significantly promoted the protein and mRNA expression of ZO-1 and occludin.Under light microscopy,the degree of lung tissue edema,alveolar hemorrhage,and inflammatory cell infiltration were significantly reduced,indicating a certain protective effect on ALI rats.The results of non-targeted metabolomics studies showed that there were 91 and 33 significantly different metabolites in the serum and lung tissue of rats treated with SLEL,respectively.Among them,the main differential metabolites in the serum were sphingosine,L-lactic acid,nicotinic acid,D-nucleotide,and mevalonate-5P,while the main differential metabolites in the lung tissue were tauro-cholic acid.This suggests that SLEL may mainly affect the metabolic pathways of sphingolipids,pyruvate,nicotinic acid,nicotinamide,and tryptophan in the serum and the metabolic pathways of taurine and hypotaurine in the lung tissue to improve ALI.Conclusion:SLEL has a significant protective effect on rats against LPS-induced ALI,and its mechanism of action may be related to the inhibition of inflammatory factors,improvement of lung barrier function,and regulation of related metabolic pathways.

Objective To explore the value of serum matrix metalloproteinase 7 combined with glutamyl transferase and total bile acids in the diagnosis of biliary atresia.Methods 112 children hospitalized in Kunming Children's Hospital with the cholestatic jaundice from July 2023 to September 2024 were selected as the research subjects.According to the surgical exploration,intraoperative cholangiography,liver biopsy and follow-up,the children were divided into the biliary atresia group(BA)(n=52)and non-biliary atresia group(Non-BA)(n=60)respecvely.The age,gender,serum matrix metalloproteinase 7(MMP-7),glutamyl transferase(GGT),alanine aminative aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TB),direct bilirubin(DB),total bile acid(TBA)and aspartate aminotransferase/platelet index(APRI)were compared between the two groups.Statistically significant indicators were included in receiver operating characteristic curve(ROC)analysis,and the area under the ROC curve(AUC)and optimal diagnostic margin(Youden index)were calculated.Results There was no significant difference in age,ALT,AST,DB,TB and APRI levels between the two groups of patients(P>0.05);There was a difference in gender composition ratio between the two groups(P=0.006);MMP-7,GGT,TBA levels in the BA group were significantly higher than those in the Non-BA group,and the difference was statistically significant(P<0.05);the AUC of MMP-7,GGT,and TBA in diagnosing BA were 0.946(95%CI 0.897～0.996),0.857(95%CI 0.789～0.926),0.654(95%CI 0.552～0.755)respectively;When the cut-off value of MMP-7 was 22.37 ng/ml,the sensitivity and specificity for diagnosing BA were 0.923 and 0.933 respectively;When the cut-off value of GGT was 151.5 U/L,the sensitivity and specificity of diagnosing BA were 0.885 and 0.733 respectively;When the cut-off value of TBA was 119.5 μmol/L,the sensitivity and specificity of diagnosing BA were 0.788 and 0.500,respectively.The AUC of MMP-7+GGT and MMP-7+TBA combined to diagnose BA were 0.971(95%CI 0.946～0.997)and 0.943(95%CI 0.889～0.996)respectively.Conclusion Serum MMP-7 has the good diagnostic value as a single indicator for diagnosing BA;MMP-7 combined with GGT is better than a single indicator for diagnosing BA;MMP-7 combined with TBA cannot improve the diagnostic efficiency.

Objective:To study the clinical features and risk factors of prognosis of neonatal appendicitis.Methods:From January 2014 to December 2022, all infants with neonatal appendicitis and received surgery in our hospital were retrospectively analyzed.Results:A total of 6 cases were enrolled, including 1 boy and 5 girls, with gestational age 36-40 weeks, birth weight 1 990~3 300 g, age of admission 5-11 d and time from illness onset to admission 0.5-4 d. All infants had abdominal distension, combined with vomiting in 4 cases, fever in 3 cases and blood in stool in 1 case. Gastrointestinal perforation was found on preoperative abdominal X-ray in 5 cases. All 6 cases received surgery and confirmed the diagnosis of appendicitis with perforation during the surgery. Appendectomy was performed without mortality. 1 case had Amyand hernia and received high ligation of the hernia sac during operation. 1 case had meningitis and was cured after 3 weeks of antibiotic treatment. 1 case developed adhesive intestinal obstruction 3 months after surgery and underwent intestinal adhesiolysis. One case developed colonic stenosis one month after surgery. The stenotic segment of the colon was resected and primary intestinal anastomosis was performed.Conclusions:Neonatal appendicitis progresses rapidly and is difficult to diagnose. The possibility of appendicitis with perforation should be considered when preoperative abdominal X-ray suggesting pneumoperitoneum. Intraoperatively, it is necessary to pay attention to the relationship between appendiceal perforation and other lesions for comprehensive treatment, and change the surgical approach accordingly.

Objective To conduct the reevaluation on the systematic evaluation of the role of Huaier Granules in the treatment of primary liver cancer to provide the practical and reliable reference basis for clini-cal decision.Methods The literatures on the systematic evaluation of Huaier granules in the treatment of pri-mary liver cancer was retrieved.The evaluation was performed by the preferred reporting items for systematic reviews and meta-analyses(PRISMA)statement,a measure tool to assess systematic reviews(AMSTAR)2 scale and the grading of recommendations assessment,development and evaluation(GRADE)system tool.Re-sults Finally,4 literatures with a total sample size of 8 040 cases were included.In 27 items with the reported quality,the proportions of complete agreement were 66.67％,62.96％,66.67％and 70.37％,respectively.AMSTAR2 scale showed that the quality of the four articles was very low.The GRADE system tool evalua-tion showed that there were 9 medium quality evidences,8 low quality evidences and 18 extremely low quality evidences.Conclusion At present,the systematic evaluation of Huaier Granules in the treatment of primary liver cancer has different degrees of defects in the integrity of the report,methodological quality,and evidence strength of outcome indicators,which still needs more high-quality studies to provide more reliable evidence-based medical evidence to guide clinical practice.

The accuracy of quantitative hormone testing has been concerned by clinical and laboratory professionals. Most of the small molecule hormones in human body have the characteristics of low concentration and similar structure, and need to be detected by high sensitivity and high specificity detection technology to achieve accurate quantification. In the past 20 years, liquid chromatography-tandem mass spectrometry (LC-MS) has gradually moved from scientific research to clinical application because of its unique sensitivity and specificity, which is considered to be a powerful supplement to traditional immunological methods, and has continuously achieved important fruits in clinical disease diagnosis and treatment. In order to further explore the application value of mass spectrometry technology in the quantitative detection of hormones, the Chinese Journal of Laboratory Medicine invited experts from the fields of clinical endocrinology and laboratory medicine to summarize their experience and opinions on the performance characteristics, challenges and future development direction of mass spectrometry technology in clinical hormone testing. Experts agreed that the advantages of mass spectrometry technology in hormone testing have been very clear, could effectively solve some clinical challenges in the diagnosis and treatment of endocrine diseases, but still faces a series of challenges such as insufficient talent reserve, low sample throughput, high operating costs and lack of standardized management.

Objective:To screen the characteristic gut microbiota and fecal metabolites related to the efficacy of oxaliplatin-based neoadjuvant chemotherapy in patients with colorectal cancer liver metastasis, to analyze the relationship between gut microbiota and fecal metabolites, and to evaluate the predictive value of relevant markers for the efficacy of neoadjuvant chemotherapy in patients with colorectal cancer liver metastasis.Methods:This is a case-control study, 34 patients with colorectal cancer liver metastasis who were treated in Qilu Hospital of Shandong University from October 2021 to July 2022 were selected as the research objects, and were divided into chemotherapy effective group (20 cases) and chemotherapy ineffective group (14 cases) according to the efficacy evaluation criteria. Logistic regression was used to construct a prediction model to screen the microbiota and metabolic markers capable of predicting the effect of chemotherapy, and the receiver operating characteristic (ROC) curve and survival analysis curve were plotted to evaluate the predictive effect of related microbiota and metabolites on the efficacy of neoadjuvant chemotherapy.Results:There was no significant difference in the α and β diversity of gut microbiota between the patients in the chemotherapy effective group and in the ineffective group (all P>0.05). In terms of species, the relative abundance of 5 species was up-regulated and 10 species were down-regulated in the chemotherapy-effective group compared with the chemotherapy-ineffective group, and the difference was statistically significant (all P<0.05), among which Prevotella salivae could effectively predict the chemotherapy effect (AUC=0.750, P=0.007), with a sensitivity of 80.0% and a specificity of 71.4%. The overall survival of patients with high abundance (17 cases) was lower than that of patients with low abundance (17 cases) ( χ 2=5.239, P=0.022). In terms of metabolites, 20 metabolites were up-regulated and 4 metabolites were down-regulated in the chemotherapy-effective group compared with the chemotherapy-ineffective group, and the difference was statistically significant (all P<0.05), among which threonine and prostaglandin F2α-1-ethanolamide could distinguish between patients who responded to chemotherapy and those who did not respond to chemotherapy (AUC=0.743, 0.707, all P<0.05), and the overall survival of patients with high levels of relative abundance (17 cases) was higher than that of patients with low levels (17 cases) ( χ 2=4.748, 5.407, all P<0.05). The Logistic regression model of Prevotella salivae and prostaglandin F2α-1-ethanolamide was obtained through screening analysis, and the ROC curve results showed that the model had a good predictive value (AUC=0.836, sensitivity: 90.0%, specificity: 78.6%), and the overall survival of patients with high predict probability (17 cases) predicted by the model was higher than that of patients with low predict probability (17 cases) ( χ 2=9.260, P=0.002). Conclusion:Prevotella salivae and prostaglandin F2α-1-ethanolamide can be used as predictive biomarkers of neoadjuvant chemotherapy for colorectal cancer liver metastasis, and the model has good clinical reference value for prognosis assessment of patients in this cohort.

Objective:To explore the antibacterial performance and mechanism of reticulated cruciform DNA nanomachines mimicking neutrophil extracellular traps (NETs).Methods:In this study, a cross-shaped DNA nanostructure was synthesized using the one-step method to form a network under the polymerization of magnesium ions. The network was used as a template for inlaying reduced Cu 0 to form reticulated DNA-templated copper nanoclusters (CuNPs). The cruciform DNA formation process was characterized by polyacrylamide gel electrophoresis (PAGE). The reticulated cruciform DNA nanomachine was characterized by atomic force microscopy (AFM) and energy dispersive spectrometer (EDS). The optimal concentrations of Cu 2+, Mg 2+and DNA in the reticulated cruciform DNA nanomachine was determined by plate coating experiments. The antibacterial performance of the reticulated cruciform DNA nanomachine against Escherichia coli ( E. coli), Staphylococcus aureus and Klebsiella pneumoniae was verified by plate coating experiments. The aggregation effect of E. coli was evaluated by crystal violet staining, and the changes on the membrane surface of E. coli were observed by scanning electron microscopy (SEM), in order to explore the antibacterial mechanism of the reticulated cruciform DNA nanomachine. Results:PAGE showed that the band migration distance of the four DNA strands was the smallest after co-incubation. AFM showed that the DNA structure was reticulated and evenly distributed, and the Cu, Mg, and P elements coexisted in the structure. The optimal concentrations of Cu 2+, Mg 2+, and DNA required for the synthesis of the reticulated cruciform DNA nanomachine were 1 mmol/L, 100 mmol/L, and 20 μmol/L, respectively. The reticulated cruciform DNA nanomachine had the ability to inhibit E. coli, Staphylococcus aureus and Klebsiella pneumoniae, with survival rates of 39.72%, 43.56% and 60.57%, respectively. The reticulated cruciform DNA nanomachine had an aggregation effect on E. coli. The surface of the bacterial film exhibited shrinkage and fractures. Conclusion:The reticulated cruciform DNA nanomachine constructed in this study can aggregate E. coli, leading to the shrinkage and fracturing of the bacterial film, and exhibiting antibacterial effects.

Tryptophan metabolism plays an important role in immunometabolism in sepsis,and the expression of indoleamine 2,3-dioxygenase 1(IDO1),the key enzyme of tryptophan metabolism,is up-regulated during sepsis.This study was designed to construct IDO1 gene knockout THP-1 cell line using CRISPR/Cas9,and to provide a cell model for studying the role of IDO1 in macrophage function.Three gRNAs were designed for the IDO1 gene,and inserted into YKO-Lentiviral gRNA plasmid respectively to construct IDO1-gRNA recombinant plasmid,which then confirmed by restriction enzyme digestion and sequencing,and packaged as Lenti-IDO1-gRNA lentivirus.THP-1 cells were infected with Cas9 lentivirus to construct a THP-1 Cas9 cell line stably expressing Cas9 protein.Then THP-1 Cas9 cells were infected with the Lenti-IDO1-gRNA lentivirus for IDO1 gene knockout,and the monoclonal cell lines were screened by limited dilution method.The efficiency of IDO1 knockout was identified by T7E1 digestion test,PCR product sequencing and Western blotting.The proliferative activity of THP-1 cells was detected by CCK8 assay;the expression of CD11b,CD68 and CD 14 in THP-1 macrophages were detected by flow cytometry;and the phagocytic function of THP-1 macrophages was detected by neutral phagocytosis test.T7E1 restriction enzyme digestion results showed that all the three gRNAs could effectively edit IDO1 gene,and gRNA2 had the highest editing efficiency.Sequencing of PCR products showed that IDO1 gene was mutated in three THP-1 IDO 1-KO monoclonal cell lines,and Western blot result showed that IDO1 protein was not expressed,suggesting THP-1 IDO1-KO cells were successfully constructed.CCK-8 proliferation assay showed that IDO1 gene knockout could inhibit the proliferation of THP-1 cells.Flow cytometry showed that IDO1 gene knockout down-regulated the expression of CD1 1b and CD68 in THP-1 macrophages,while up-regulated the expression of CD 14.The results of neutral red phagocytosis test showed that IDO1 gene knockout promoted the phagocytosis of macrophages.In conclusion,the IDO1 gene knockout THP-1 cell line has successfully constructed by CRISPR-Cas9,and IDO1 plays an important role in regulating the proliferation activity and differentiation of THP-1 cells,as well as the phagocytic function of THP-1 macrophages.

Efforts have been made to establish various human pluripotent stem cell lines. However, such methods have not yet been duplicated in non-human primate cells. Here, we introduce a multiplexed single-cell sequencing technique to profile the molecular features of monkey pluripotent stem cells in published culture conditions. The results demonstrate suboptimized maintenance of pluripotency and show that the selected signaling pathways for resetting human stem cells can also be interpreted for establishing monkey cell lines. Overall, this work legitimates the translation of novel human cell line culture conditions to monkey cells and provides guidance for exploring chemical cocktails for monkey stem cell line derivation.
Animals ; Haplorhini ; Single-Cell Gene Expression Analysis ; Pluripotent Stem Cells/metabolism* ; Cell Line ; Signal Transduction ; Cell Differentiation ; Transcriptome