Objective To explore the differences of lipid metabolites in tongue coating between ischemic stroke patients and healthy people,thus to screen out the potential biomarkers for tongue coating of ischemic stroke patients.Methods Twenty patients confirmed as ischemic stroke were selected as the observation group,and another 20 controls receiving healthy medical checkup at the same period were selected as the control group.All of them were aged between 40 and 80 years.Using ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS)technology,and by combined with the statistical methods of principal component analysis(PCA)and partial least squares discriminant analysis(PLS-DA),non-targeted lipid metabolomics study was performed on tongue coating samples of the observation group and the control group.According to the criteria that the VIP value in the orthogonal signal correction-based orthogonal partial least squares discriminant analysis(OPLS-DA)model was greater than 1 and the P value of the Wilcoxon rank test was less than 0.05,lipid metabolites with significant differences were screened and determined.Results Differential lipid metabolites exist in ischemic stroke patients(observation group)and healthy people(control group),and a total of 14 lipid metabolites were identified,and they were free fatty acids,ceramide,sphingomyelin,cardiolipin,lecithin,etc.Of the 14 lipid metabolites,4 were down-regulated and 10 were up-regulated,and the differences were statistically significant(P＜0.05).Conclusion There exist differential lipid metabolites in tongue coating of patients with ischemic stroke and the healthy controls,and these different lipid metabolites are expected to become potential biomarkers for ischemic stroke.

Objective To explore the role and potential mechanism of Astragali Radix-derived extracelluar vesicles(EVs)-like particles(AR-EVLP)in diabetic wound healing,providing a novel therapeutic drug mode and theoretical basis for traditional Chinese medicine in treating diabetic skin ulcers.Methods AR-EVLP was extracted using ultracentrifugation.Untargeted metabolomics was used to analyze the potential active components of AR-EVLP,and the main active substances in AR-EVLP were identified by comparison with the Herb database.The targets of the active components were obtained through the Swiss Target Prediction database and the TCMSP database.Targets related to diabetic wound healing were obtained from the GeneCards database.Key targets were identified by intersecting drug targets and disease targets.GO functional annotation and KEGG pathway enrichment analysis were performed using the DAVID database.Based on the pathway enrichment results,a"drug components-targets-pathways"diagram was constructed to identify core targets.Molecular docking and visualization were performed using Autodock and PyMOL software.Results Formononetin was identified as the main active component in AR-EVLP,with 66 key targets related to diabetes and wound healing,including 15 key node proteins such as SRC,CASP3 and JUN.GO functional enrichment analysis suggested that formononetin can regulate biological processes such as protein phosphorylation and gene expression.KEGG pathway enrichment analysis indicated potential involvement in multiple signaling pathways,including VEGF and TNF.Seven targets,including PIK3CA,JUN and MAPK14,were identified as core targets for formononetin.Molecular docking showed that formononetin had the strongest binding affinity with MAPK14.Conclusion AR-EVLP may be a potential effective drug mode for the treatment of diabetic wound healing.

Objective:To construct a calcium alginate (ALG-Ca 2+) composite hydrogel loaded with chlorella protein (Cp) (ALG-Ca 2+@Cp) and investigate its combined effect with microwave hyperthermia on the proliferation, apoptosis, and immune activation of mouse pancreatic cancer cells. Methods:ALG-Ca 2+@Cp was prepared using a physical cross-linking method and its physiochemical properties was characterized via scanning electron microscopy, rheological analysis, Ca 2+ release experiments, and microwave thermal conversion tests. The BCA protein quantification assay was used to evaluate the adsorption capacity of ALG-Ca 2+@Cp for pancreatic cancer cell antigens. The effects of ALG-Ca 2+@Cp extract combined with microwave intervention on pancreatic cancer cell proliferation, apoptosis protein expression, and cell viability were assessed using CCK-8 assays, ELISA, and Calcein-AM/PI double fluorescence staining. Flow cytometry was performed to determine the maturation-promoting ability of ALG-Ca 2+@Cp on immature mouse bone marrow-derived dendritic cells (BMDCs). Results:ALG-Ca 2+@Cp exhibited a three-dimensional network structure with a storage modulus (G') greater than the loss modulus (G''), demonstrating typical hydrogel properties. The hydrogel loaded with 0.5 mol/ml Ca 2+ reached 48°C after 5 minutes of microwave irradiation at 5.0 W/cm 2, and Ca 2+ release plateaued within 5 minutes. ALG-Ca 2+@Cp effectively adsorbed pancreatic cancer cell antigens. Combined with microwave treatment, it significantly reduced pancreatic cancer cell proliferation ( A450 value 0.39±0.07 vs 2.78±0.15) and increased apoptosis markers calreticulin (CRT) and high mobility group box-1 protein (HMGB1) [(557.09±37.84) pg/ml vs (135.14±11.84) pg/ml, (4.77±0.18) ng/ml vs (1.6±0.16) ng/ml], leading to decreased cell viability; and all the differences were statistically significant (all P value <0.05). ALG-Ca 2+@Cp synergistically promoted the maturation of immature BMDCs in the presence of pancreatic cancer antigens, with a CD 80+ positivity rate of (75.67±6.53)%. Conclusions:ALG-Ca 2+@Cp is successfully constructed. Its combination with microwave hyperthermia can significantly enhance the cytotoxicity and immune activation against mouse pancreatic cancer cells by targeting intracellular antigens and inducing immunogenic cell death.

Objective:To construct a calcium alginate (ALG-Ca 2+) composite hydrogel loaded with chlorella protein (Cp) (ALG-Ca 2+@Cp) and investigate its combined effect with microwave hyperthermia on the proliferation, apoptosis, and immune activation of mouse pancreatic cancer cells. Methods:ALG-Ca 2+@Cp was prepared using a physical cross-linking method and its physiochemical properties was characterized via scanning electron microscopy, rheological analysis, Ca 2+ release experiments, and microwave thermal conversion tests. The BCA protein quantification assay was used to evaluate the adsorption capacity of ALG-Ca 2+@Cp for pancreatic cancer cell antigens. The effects of ALG-Ca 2+@Cp extract combined with microwave intervention on pancreatic cancer cell proliferation, apoptosis protein expression, and cell viability were assessed using CCK-8 assays, ELISA, and Calcein-AM/PI double fluorescence staining. Flow cytometry was performed to determine the maturation-promoting ability of ALG-Ca 2+@Cp on immature mouse bone marrow-derived dendritic cells (BMDCs). Results:ALG-Ca 2+@Cp exhibited a three-dimensional network structure with a storage modulus (G') greater than the loss modulus (G''), demonstrating typical hydrogel properties. The hydrogel loaded with 0.5 mol/ml Ca 2+ reached 48°C after 5 minutes of microwave irradiation at 5.0 W/cm 2, and Ca 2+ release plateaued within 5 minutes. ALG-Ca 2+@Cp effectively adsorbed pancreatic cancer cell antigens. Combined with microwave treatment, it significantly reduced pancreatic cancer cell proliferation ( A450 value 0.39±0.07 vs 2.78±0.15) and increased apoptosis markers calreticulin (CRT) and high mobility group box-1 protein (HMGB1) [(557.09±37.84) pg/ml vs (135.14±11.84) pg/ml, (4.77±0.18) ng/ml vs (1.6±0.16) ng/ml], leading to decreased cell viability; and all the differences were statistically significant (all P value <0.05). ALG-Ca 2+@Cp synergistically promoted the maturation of immature BMDCs in the presence of pancreatic cancer antigens, with a CD 80+ positivity rate of (75.67±6.53)%. Conclusions:ALG-Ca 2+@Cp is successfully constructed. Its combination with microwave hyperthermia can significantly enhance the cytotoxicity and immune activation against mouse pancreatic cancer cells by targeting intracellular antigens and inducing immunogenic cell death.

Circulating tumor cells (CTC) and circulating tumor DNA(ctDNA)are cancer cells that detach from their primary site to circularity system or other body fluids,which hold the promise of monitoring of tumor evolution and therapeutic efficacy and improving prognosis for patients.With the rapid development of detection technologies of CTCs and ctDNA,the sensitivity and specificity of current assays had significant improved.CTCs can be detected by use of cytometry,immunofluorescence analysis and genedetection and the detection methods of ctDNA mainly based on PCR and the next generation sequencing.As the developing of various methods and platforms were developed for detecting the circulating tumor cells and circulating tumor DNA,it is necessary to use the detection technology flexibly according to the research purpose and provide more assistance for the diagnosis and treatment of tumor.

Objective: Alport syndrome was an inherited kidney disease caused by the mutation of COL4A3, COL4A4, or COL4A5. Whole-exome sequencing was used to detect the mutations on these genes for the molecular diagnosis of Alport syndrome. Methods: A 6-year-old girl found accidentally with microscopic hematuria at the age of 4. The clinical data and blood sample of the family including proband, parents, brothers, and sisters were collected. Whole exome sequencing was conducted using their genomic DNAs. Results: A novel heterozygous frameshift mutation c.1826delC (p.Pro609Glnfs*44) was found in the exon 25 of the COL4A4 (NM_000092) in the proband, the father, and the sister, showing an autosomal dominant inheritance pattern of Alport syndrome. This mutation of COL4A4 was confirmed by mutation analysis, and the mutation of c.1826delC was verified by Sanger sequencing. No mutations on COL4A3 and COL4A5 were detected in this family. And the mother and brother are normal wide-type. Conclusions This novel mutation is a valuable addition to the current genetic profile of Alport syndrome, and provide us a better understanding of the disease. Whole-exome sequencing is a power tool to identify the novel mutations of inherited disease and contribute to the molecular diagnosis of disease.