Objective:To analyzed the chemical components of Black Tartary Buckwheat using Ultra-performance liquid chromatography coupled with quadrupole-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-Q-Exactive Orbitrap MS); To further compared the compositional differences under various drying conditions.Methods:UPLC-Q-Exactive Orbitrap MS was employed to scan 14 samples of Black Tartary Buckwheat in 4 batches, aiming to identify the major chemical components. ACQUITY UPLC HSS T3 C18 column (2.1 mm×100 mm, 1.7 μm) was adopted; mobile phase was 0.1% formic acid aqueous solution-acetonitrile solution; flow rate was 0.3 ml/min for gradient elution; HESI-Ⅱ ion source was used to detect negative ions. According to general mass spectroscopy rules and literature, at the same time, Compound Discoverer 3.2 and MSConvert software were used to analyze the data, upload to GNPS network, and finally generate network diagram using Cytocape 3.6.1 software.Results:A total of 134 components were identified in Black Tartary Buckwheat, of which 76 compounds were identified in the negative ion mode, 79 compounds were identified in the positive ion mode, and 21 chemicals were simultaneously identified as positive and negative ions. Based on the similarity cluster analysis of UPLC-Q-Exactive Orbitrap MS secondary mass spectrometry fragment patterns, a molecular network was established, and the main compounds in Black Tartary Buckwheat were flavonoids, phenolic acids and amino acids, and the results showed that the positive and negative ion modes had similar clustering results. There were certain differences in the content of flavonoids, phenolic acids, and amino acids in Black Tartary Buckwheat under two different drying conditions.Conclusion:The established UPLC-Q-Exactive Orbitrap MS method enables rapid identification of the main chemical components in Black Tartary Buckwheat, while the constructed molecular network provides a reference framework for further research on its chemical composition. Additionally, the drying conditions are found to exert certain effects on the content of these chemical components.

Objective To analyze the expression levels and clinical significance of N-myc down-stream-regulated gene 3(NDRG3)and semaphoring 3A(SEMA3A)in elderly patients with acute ischemic stroke(AIS).Methods A total of 100 elderly AIS patients admitted to Department of Geriatrics of Liaocheng People's Hospital from September 2020 to September 2022 were included as the study group.According to their NIHSS score at admission,they were divided into mild(34 cases),moderate(31 cases)and severe(35 cases)subgroups.All patients were followed up for 3 months after discharge.And they were assigned into good prognosis group(69 cases)and poor prognosis group(31 cases)based on the modified Rankin scale score.Another 100 healthy individ-uals who underwent physical examination in our hospital during the same period were recruited as the control group.Western blotting was used to detect the protein expression of NDRG3 and SEMA3A in peripheral blood mononuclear cells(PBMC).ELISA was applied to measure the con-tents of VEGF,TGF-1,TNF-α,and IL-17 in peripheral blood samples.Spearman rank correlation analysis was performed to analyze the correlation of NDRG3 and SEMA3A levels with NIHSS score,and ROC curve was plotted to analyze the values of NDRG3 and SEMA3A in predicting poor prognosis in elderly AIS patients.Results The expression levels of NDRG3 and SEMA3A in PBMC were obviously higher in the study group than the control group(1.11±0.16 vs 0.76± 0.13,0.78±0.13 vs 0.42±0.09,P＜0.01).The levels in the mild,moderate and severe subgroups were significantly higher than that of the control group(P＜0.01).The poor prognosis group had statistically higher expression levels of NDRG3 and SEMA3A than the good prognosis group(P＜0.01).Spearman rank correlation analysis showed that the NIHSS score was positively corre-lated with the expression levels of NDRG3 and SEMA3A in elderly AIS patients(r=0.597,P＜0.01;r=0.618,P＜0.01),while the NDRG3 level was positively correlated with that of SEMA3 A(r=0.477,P＜0.01).ROC curve analysis indicated that the AUC value of combined NDRG3 and SEMA3A levels was superior to that of NDRG3 and SEMA3A alone in predicting of poor progno-sis(0.962 vs 0.861,0.880,P＜0.01).Conclusion The levels of NDRG3 and SEMA3A proteins are up-regulated in elderly AIS patients,and are closely associated with the severity and prognosis of the disease.

Objective:To investigate the expression and significance of Nek2B and β-catenin expression in triple negative breast cancer (TNBC) at molecule levels.Methods:By using the methods of bioinformatics [GEO2R online tool, gene ontology (GO) function analysis, KEGG biological pathway enrichment analysis], the differentially expressed genes were screened from TNBC microarray data.Expression levels of Nek2B and β-catenin TNBC cell lines were detected by Western blot and qRT-PCR.From January 1, 2007 to December 31, 2012, eighty cases of TNBC were collected from the Second Hospital of Shanxi Medical University. The expression of Nek2B in TNBC tumor tissue was detected by immunohistochemistry and tissue microarray, and the relationship between Nek2B and clinical pathological characteristics of TNBC was analyzed.Results:Through bioinformatics analysis of the cDNA chip sets of 2 TNBC tumors(GSE38959,GSE27447), 998 differentially expressed genes were obtained in the initial screening, and 13 differentially expressed genes were revealed after intersection. The results of biological pathway analysis showed that the common differential expression genes were closely related to Wnt/β-catenin pathway, among which Nek2 expression showed the greatest difference and was associated with poor prognosis. Expression intensity of Nek2B and repeated β-catenin in the same TNBC cell line was consistent.The results of immunohistochemistry showed that the high expression of Nek2B was related to the high histological stage (G3;84.3% vs.37.9%, P<0.001), lymph node metastasis group (76.7% vs.54.1%, P=0.032), high Ki-67 positive index group (78.6% vs.52.6%, P=0.007) and β-catenin positive expression group (72.5% vs.27.3%, P=0.018). Conclusions:The high level of Nek2B expression is related to a poor prognosis in TNBC patients. In TNBC tissues and cells, the expression of Nek2B is correlated with β-catenin, suggesting that Nek2B may affect the occurrence and development of TNBC by regulating the Wnt/β-catenin patients signaling pathway.