Objective To explore the roles of transcription factor E26 transformation-specific 1(ETS1)and F-box protein 45(FBXO45)in invasion and metastasis in hepatocellular carcinoma cells and the potential molecular mechanism.Methods Jaspar,hTFtarget and Cistrome transcription factor database prediction websites were used to predict the transcription factors of FBXO45.According to the intersection of the predicted results of each database,the expression of FBXO45 was detected after the candidate transcription factors were knockdown in HCCLM3 and Huh7 liver cancer cells,respectively.The most significant influence on FBXO45 expression was selected for further analysis,and chromatin immunoprecipitation assay(ChIP)was used to verify the binding to the FBXO45 promoter.Finally,the potential transcription factor of FBXO45 was identified.The effect of ETS1 overexpression on invasion and migration in HCCLM3 and Huh7 cells was detected by Transwell assay,and the expression levels of epithelial-mesenchymal transition(EMT)pathway proteins were detected by Western blot assay.The effects of FBXO45 knockdown on the invasion and migration under the condition of overexpression of ETS1 were also studied.Results Intersection of FBXO45 transcription factors identified 3 candidate transcription factors,ETS1,SPI1 and YY1.When the 3 transcription factors were knocked down in HCCLM3 and Huh7 cells,respectively,ETS1 knockdown significantly reduced the expression of FBXO45.According to the analysis of The Cancer Genome Atlas(TCGA)data and the Gene Expression Omnibus(GEO)data,the expression levels of ETS1 and FBXO45 were significantly positively correlated(R=0.31,P<0.000 1;R=0.40,P=0.021 9).ChIP suggested that ETS1 could specifically bind to FBXO45 promoter sequence to regulate its expression,confirming that ETS1 was a potential transcription factor of FBXO45.After overexpression of ETS1 in HCCLM3 and Huh7 cells,the invasion and migration abilities of cells were significantly enhanced,and the expression of N-cadherin and Snail was up-regulated(P<0.01).In addition,in the case of ETS1 overexpression,FBXO45 knockdown significantly inhibited the invasion and migration(P<0.01).Conclusion ETS1 activates the transcription of FBXO45 and leads its high expression,which enhances the invasion and migration of HCC cells via EMT pathway and promotes the progression of hepatocellular carcinoma.

Objective To investigate the ameliorative effect of Qihuakeli,a Hani formula,on glycolipid metabolism in type 2 diabetes mellitus by in vivo and in vitro experiments.Methods Rat liver mesenchymal stromal cells(BRL-3A)were inoculated in six-well plates and divided into blank,palmitic acid,fenofibrate,and Qihuakeli serum-containing 5.4,10.8,and 21.6 g/kg groups.Except for the blank group,the remaining groups were intervened with 0.2 mmol/L palmitic acid(PA)for 24 hour,and then added with drug-containing serum,and then continued to incubate for 24 hour.The proliferation rate of BRL-3A cells in each group was determined.Total cholesterol(T-CHO),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)concentrations in the supernatant of each cell group were measured,cell culture medium was aspirated and discarded,triglyceride(TG)concentration in the cell lysate.The lipid content of the cells was determined by measuring and staining with red oil.Meanwhile,45 rats were taken and divided into blank group,model group,fenofibrate group(0.225 g/kg),Qihuakeli compound 5.4 g/kg group,and Qihuakeli compound 10.8 g/kg group,the blank group was given normal feed and the rest of the groups were given high-fat feed for 42 day.Beginning on the 43rd day,each group,except the blank group,was injected with a single intraperitoneal injection of Starting from the 43rd day,except the blank group,each group was given a one-time intraperitoneal injection of 0.25%streptozotocin(STZ)solution,and at the same time,the corresponding drugs were given by gavage for 14 day.The rats'weight gain and liver index were measured.Serum fasting blood glucose(FBG)and fasting insulin(FINS)were detected,and the insulin resistance index(ISI)was calculated.Serum free fatty acid(FFA)levels and tumor necrosis factor-α(TNF-α)in liver tissue were also detected.HE staining was used to detect pathological changes in the pancreas.Pathological changes were observed in the tissues,and islet α and β cell expression was detected by immunohistochemistry.Results Compared to the PA group,the accumulation rate of BRL-3A cells was significantly higher(P<0.01)in the 10.8 and 21.6 g/kg Qihuakeli-containing serum groups.The levels of T-CHO,LDL-C and TG in the 5.4 and 21.6 g/kg serum groups were significantly lower(P<0.05),and HDL-C levels significantly increased(P<0.05).Oil red staining results showed that lipids in the cytoplasm of the 5.4,10.8 and 21.6 g/kg.Qihuacel-containing groups significantly reduced.Compared to the model group,the body weight of the 10.8 g/kg group containing Qihuakeli granules increased significantly(P<0.05).The liver index of the 5.4 g/kg group containing Qihuakeli decreased significantly(P<0.05).The serum indices of FBG,FINS,FFA and insulin resistance of the 5 g/kg group containing Qihuakeli decreased significantly(P<0.05).In the 5.4,10.8 g/kg groups,all serum FBG,FINS,FFA and insulin resistance indices significantly reduced in the 5.4 and 10.8 g/kg Qihuakeli groups(P<0.05 or P<0.01).TNF-α levels were significantly reduced(P<0.01).HE staining showed that a small number of lymphocytes were scattered in the pancreatic ducts and perivascular area of the rats in the Qihuakeli 5.4 and 10.8 g/kg groups,the local vasodilatation was observed,the number of pancreatic islet cells and the area of islet cells significantly increased.Immunohistochemical study was further used.The results of immunohistochemistry showed that the area of pancreatic islet α-cells significantly reduced and the area of pancreatic islet β-cells significantly increased in Qihuakeli 5.4 and 10.8 g/kg groups.Conclusion Qihuakeli compound improved glucose-lipid metabolism in T2DM,probably by improving the function of pancreatic islet cells,increasing the sensitivity of insulin to blood glucose,improving insulin resistance,decreasing the secretion of insulin and glucagon,and thus lowering the level of fasting blood glucose.Meanwhile,by decreasing the content of TNF-α,inhibiting lipolysis in the body,and promoting the uptake of FFA by adipocytes,and further lowering the FFA.Thus,it regulates the levels of TG,T-CHO,HDL-C and LDL-C,improves the abnormalities of glucose and lipid metabolism,and alleviates T2DM.

Objective To investigate the ameliorative effect of Qihuakeli,a Hani formula,on glycolipid metabolism in type 2 diabetes mellitus by in vivo and in vitro experiments.Methods Rat liver mesenchymal stromal cells(BRL-3A)were inoculated in six-well plates and divided into blank,palmitic acid,fenofibrate,and Qihuakeli serum-containing 5.4,10.8,and 21.6 g/kg groups.Except for the blank group,the remaining groups were intervened with 0.2 mmol/L palmitic acid(PA)for 24 hour,and then added with drug-containing serum,and then continued to incubate for 24 hour.The proliferation rate of BRL-3A cells in each group was determined.Total cholesterol(T-CHO),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)concentrations in the supernatant of each cell group were measured,cell culture medium was aspirated and discarded,triglyceride(TG)concentration in the cell lysate.The lipid content of the cells was determined by measuring and staining with red oil.Meanwhile,45 rats were taken and divided into blank group,model group,fenofibrate group(0.225 g/kg),Qihuakeli compound 5.4 g/kg group,and Qihuakeli compound 10.8 g/kg group,the blank group was given normal feed and the rest of the groups were given high-fat feed for 42 day.Beginning on the 43rd day,each group,except the blank group,was injected with a single intraperitoneal injection of Starting from the 43rd day,except the blank group,each group was given a one-time intraperitoneal injection of 0.25%streptozotocin(STZ)solution,and at the same time,the corresponding drugs were given by gavage for 14 day.The rats'weight gain and liver index were measured.Serum fasting blood glucose(FBG)and fasting insulin(FINS)were detected,and the insulin resistance index(ISI)was calculated.Serum free fatty acid(FFA)levels and tumor necrosis factor-α(TNF-α)in liver tissue were also detected.HE staining was used to detect pathological changes in the pancreas.Pathological changes were observed in the tissues,and islet α and β cell expression was detected by immunohistochemistry.Results Compared to the PA group,the accumulation rate of BRL-3A cells was significantly higher(P<0.01)in the 10.8 and 21.6 g/kg Qihuakeli-containing serum groups.The levels of T-CHO,LDL-C and TG in the 5.4 and 21.6 g/kg serum groups were significantly lower(P<0.05),and HDL-C levels significantly increased(P<0.05).Oil red staining results showed that lipids in the cytoplasm of the 5.4,10.8 and 21.6 g/kg.Qihuacel-containing groups significantly reduced.Compared to the model group,the body weight of the 10.8 g/kg group containing Qihuakeli granules increased significantly(P<0.05).The liver index of the 5.4 g/kg group containing Qihuakeli decreased significantly(P<0.05).The serum indices of FBG,FINS,FFA and insulin resistance of the 5 g/kg group containing Qihuakeli decreased significantly(P<0.05).In the 5.4,10.8 g/kg groups,all serum FBG,FINS,FFA and insulin resistance indices significantly reduced in the 5.4 and 10.8 g/kg Qihuakeli groups(P<0.05 or P<0.01).TNF-α levels were significantly reduced(P<0.01).HE staining showed that a small number of lymphocytes were scattered in the pancreatic ducts and perivascular area of the rats in the Qihuakeli 5.4 and 10.8 g/kg groups,the local vasodilatation was observed,the number of pancreatic islet cells and the area of islet cells significantly increased.Immunohistochemical study was further used.The results of immunohistochemistry showed that the area of pancreatic islet α-cells significantly reduced and the area of pancreatic islet β-cells significantly increased in Qihuakeli 5.4 and 10.8 g/kg groups.Conclusion Qihuakeli compound improved glucose-lipid metabolism in T2DM,probably by improving the function of pancreatic islet cells,increasing the sensitivity of insulin to blood glucose,improving insulin resistance,decreasing the secretion of insulin and glucagon,and thus lowering the level of fasting blood glucose.Meanwhile,by decreasing the content of TNF-α,inhibiting lipolysis in the body,and promoting the uptake of FFA by adipocytes,and further lowering the FFA.Thus,it regulates the levels of TG,T-CHO,HDL-C and LDL-C,improves the abnormalities of glucose and lipid metabolism,and alleviates T2DM.

Objective To investigate the clinical application value of predicting the severity of acute pancreatitis(AP)based on MRI-T2WI radiomics nomogram.Methods A total of 375 patients with AP were analyzed retrospectively,who were divided into 281 cases in the training group and 94 cases in the validation group according to the ratio of 3∶1.Based on MRI-T2WI image,man-ual segmentation was performed for the pancreatic parenchyma.The radiomics feature were selected by feature extraction and dimen-sionality reduction,the support vector machine(SVM)classifier were used to construct the radiomics model.Logistic regression analysis was used to screen out independent risk factors,and an radiomics nomogram model was constructed in combined with the Radiomics score(Radscore),and the predictive performances of the models were evaluated.Results Receiver operating characteristic(ROC)curve analysis showed that the predictive efficacy of radiomics nomogram model[training group,area under the curve(AUC)=0.893;val-idation group,AUC=0.889]was higher than that of clinical model(training group,AUC=0.799;validation group,AUC=0.809)and radiomics model(training group,AUC=0.814;validation group,AUC=0.823).Conclusion The radiomics nomogram based on MRI-T2WI radiomics features and independent risk factors has high clinical application value for the prediction of AP severity.

Objective: To analyze the characteristics of trigeminal event-related potentials (tERPs) in different kinds of olfactory disorders (OD), and to evaluate the importance of tERPs for the patients with olfactory dysfunction. Methods: Clinical data of 314 patients with olfactory dysfunction from the Smell and Taste Clinics in Beijing Anzhen Hospital from 2015 to 2021 were retrospectively reviewed, including 158 males and 156 females, aging from 6 to 78 years. The control group consisted of healthy people from medical examination center, who were gender and age matched. The clinical characteristics of OD were analyzed using Sniffin' Sticks test, olfactory event-related potentials (oERPs), tERPs and acoustic rhinometry test. SPSS 17.0 software was used to compare the difference of tERPs between the two groups, and to analyze the related factors affecting trigeminal function. Results: The ratio of tERPs presence was different in OD caused by different reasons: head traumatic OD (54.9%), post-virus infection OD (63.6%), sinonasal inflammatory OD (68.4%) and OD due to other causes (56.9%). Compared with controls, tERPs signals in OD patients showed a significant lower amplitude in the N₁ wave (all P<0.001), and lower amplitude in the P₂ wave in most OD patients (head trauma t=-4.11, P<0.001; sinonasal inflammation t=-2.04, P=0.046; others t=-2.40, P=0.020) except in OD by post-virus infection (t=-1.98, P=0.052). tERPs signals in OD patients by sinonasal inflammation showed longer latency in the N₁ wave (t=2.15, P=0.036), but this difference was not observed in other OD patients (all P>0.05). tERPs signals were significantly correlated with the Sniffin' Sticks score, deficiency of oERPs and nasal minimum cross-sectional area (all P<0.05). Conclusions: OD patients show neurophysiologic deficits in trigeminal function. The absence of tERPs or lower amplitude in N₁ waves are the important characteristics of patients with OD.
Evoked Potentials/physiology* ; Female ; Humans ; Inflammation ; Male ; Olfaction Disorders/etiology* ; Retrospective Studies ; Smell/physiology* ; Virus Diseases/complications*

Biological sample pretreatment is an important step in biological sample analysis. Due to the diversity of biological matrices, the analysis of target substances in these samples presents significant challenges to sample processing. To meet these emerging demands on biopharmaceutical analysis, this paper summarizes several new techniques of on-line biological sample processing: solid phase extraction, solid phase micro-extraction, column switching, limited intake filler, molecularly imprinted solid phase extraction, tubular column, and micro-dialysis. We describe new developments, principles, and characteristics of these techniques, and the application of liquid chromatography-mass spectrometry (LC-MS) in biopharmaceutical analysis with these new techniques in on-line biological sample processing.

Zidovudine (AZT), the first drug approved by the US Food and Drug Administration for the treatment of human immunodeficiency virus (HIV) infection, is metabolized in the host cells to 5'-AZT triphosphate (AZT-TP) which inhibits HIV reverse transcriptase. As the pharmacokinetics of AZT and its phosphorylated metabolites in human peripheral blood mononuclear cells (hPBMCs) is limited, the aim of this study was to determine the pharmacokinetic parameters of AZT and its phosphorylated metabolites in hPBMCs from 12 healthy Chinese male subjects after a single oral dose of 600 mg of AZT. Blood samples were collected prior to drug administration, then at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8 and 10 h after drug administration. Mononuclear cells collected by Ficoll-Hypaque density gradient centrifugation were used for determination of AZT and metabolites [AZT monophosphate (AZT-MP), AZT diphosphate (AZT-DP) and AZT-TP] and the plasma was used to evaluate the pharmacokinetics of AZT. Plasma concentration of AZT peaked within 0.583 h and intracellular concentrations of AZT, AZT-MP, AZT-DP and AZT-TP peaked within 1.083, 1.500, 1.417 and 1.583 h, respectively. AZT in plasma was eliminated rapidly with t 1/2 of 2.022 h, and AZT-MP, AZT-DP and AZT-TP were eliminated with t 1/2 of 13.428, 8.285 and 4.240 h, respectively. The plasma concentration of the phosphorylated metabolites was not quantifiable.