Objective:To investigate the relations of serum hypoxia inducible factor 2α (HIF-2α) and miR-21 expressions with cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage (aSAH) after interventional embolization.Methods:One hundred and seventy-four patients with aSAH underwent interventional embolization in our hospital from October 2017 to June 2019 were prospectively selected. DSA examination was performed 4 d after surgery, and severity of cerebral vasospasm was evaluated. Enzyme-linked immunosorbent assay was used to detect the level of serum HIF-2α and reverse transcription-PCR was employed to detect the serum miR-21 expression before and 3 and 7 d after interventional embolization. The clinical data and changes of serum HIF-2α and miR-21 expressions in patients with different degrees of cerebral vasospasm were compared. Correlation analysis was performed to analyze the relation of HIF-2α expression with miR-21 expression 3 and 7 d after interventional embolization. Receiver operating characteristics curve was used to analyze the diagnostic values of serum HIF-2α and miR-21 levels in cerebral vasospasm 3 d after interventional embolization.Results:There were 100 patients without vasospasm, and 20, 38 and 16 patients with mild, moderate and severe cerebral vasospasm, respectively. The serum levels of HIF-2α and miR-21 in patients with mild, moderate and severe cerebral vasospasm increased successively 3 and 7 d after interventional embolization, with significant differences ( P<0.05). Positive correlation was noted between expressions of HIF-2α and miR-21 in serum 3 and 7 d after interventional embolization ( P<0.05). Area under the curve (AUC) of HIF-2α in diagnosis of cerebral vasospasm was 0.748 ( 95%CI: 0.615-0.883, P=0.000) 3 d after interventional embolization. AUC of serum miR-21 level in diagnosis of cerebral vasospasm was 0.715 ( 95%CI: 0.590-0.842, P=0.000). AUC of serum HIF-2α combined with miR-21 in diagnosis of cerebral vasospasm was 0.893 ( 95%CI: 0.792-0.985, P=0.000). When diagnostic critical points of HIF-2α and miR-21 were 82.75 pg/mL and 1.15, the sensitivity, accuracy and negative predictive value of HIF-2α combined with miR-21 in the diagnosis of cerebral vasospasm were higher than those of HIF-2α or miR-21 alone. Conclusion:The expressions of serum HIF-2α and miR-21 in patients with aSAH after interventional embolization can effectively predict the occurrence of cerebral vasospasm, and may be involved in the occurrence and development of cerebral vasospasm.

OBJECTIVETo establish an efficient and high resolution 2-DE (two-dimensional electrophoresis) protocol for root tuber proteome analysis of Rehmannia glutinosa.

METHODProteins from root tuber of R. glutinosa were extracted by using five different methods and their productivity and profiles were assessed by means of SDS-PAGE and two-dimensional electrophoresis.

RESULTThe trichloroacetic acid (TCA)-phenol extraction method was found most effective for the extraction with the highest protein yield, the most spots in protein patterns, and the highest resolution of proteins, and the clearest background could be achieved simultaneously. A 1:5 solution of ampholine pH 3-10 and pH 5-8 for a nonlinear gel and the 170 microg of protein loading dosage obtained maps with more protein spots and higher resolution of separation patterns.

CONCLUSIONThis study based on the optimized root tuber proteome preparation and the 2-DE protocol gets a high resolution and reproducibility 2-DE image, which will be expected to have excellent applications in proteomics studies of R. glutinosa tuber root.

Electrophoresis, Gel, Two-Dimensional ; methods ; Plant Proteins ; analysis ; chemistry ; Plant Roots ; chemistry ; Proteome ; chemistry ; isolation & purification ; Rehmannia ; chemistry

Objective To establish a new model of indocyanine green (ICG) clearance test combined with total bilirubin actual resident rate (TBARR) for predicting the short-term prognosis of patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) treated with artificial liver support system (ALSS) therapy. Methods A retrospective analysis was performed for the clinical data of 136 patients with HBV-ACLF who underwent ALSS therapy in Department of Infectious Diseases, The Affiliated Hospital of Southwest Medical University, from June 2017 to July 2021, and according to the prognosis at 3-month follow-up, they were divided into survival group with 92 patients and death group with 44 patients. Related indicators were measured at the time of the confirmed diagnosis of ACLF, including biochemical parameters, coagulation, indocyanine green retention rate at 15 minutes (ICGR 15 ), and effective hepatic blood flow (EHBF), and related indices were calculated, including Model for End-Stage Liver Disease (MELD) score, MELD difference (ΔMELD), Child-Turcotte-Pugh (CTP) score, total bilirubin clearance rate (TBCR), total bilirubin rebound rate (TBRR), and TBARR. The Mann-Whitney U test was used for comparison of continuous data with skewed distribution between two groups; the chi-square test was used for comparison of categorical data between groups. A binary logistic regression analysis was used to establish a combined predictive model for the prognosis of HBV-ACLF after ALSS therapy. The area under the ROC curve (AUC) was used to compare the accuracy of various models in judging the short-term prognosis of patients with HBV-ACLF after ALSS therapy, and the Z test was used for comparison of AUC. Results There were significant differences between the death group and the survival group in MELD score, ΔMELD, CTP score, ICGR 15 , EHBF, TBRR, TBARR, neutrophil count, percentage of neutrophils, lymphocyte count, platelet count, alkaline phosphatase, gamma-glutamyl transpeptidase, total bilirubin, albumin, prothrombin time, international normalized ratio, prothrombin time activity, prealbumin, fibrinogen, serum sodium, age, and the incidence rate of hepatic encephalopathy (all P < 0.05). The multivariate logistic regression analysis showed that age (odds ratio [ OR ]=1.096, 95% confidence interval [ CI ]: 1.056-1.137, P < 0.001), neutrophil count ( OR =1.214, 95% CI : 1.044-1.411, P =0.012), TBRR ( OR =0.989, 95% CI : 0.982-0.996, P =0.001), TBARR ( OR =1.073, 95% CI : 1.049-1.098, P < 0.001), ΔMELD ( OR =1.480, 95% CI : 1.288-1.701, P < 0.001), CTP score ( OR =2.081, 95% CI : 1.585-2.732, P < 0.001), and ICGR 15 ( OR =1.116, 95% CI : 1.067-1.168, P < 0.001) were independent influencing factors for short-term mortality in patients with HBV-ACLF after ALSS therapy. The binary logistic regression analysis was used to establish four combined predictive models for predicting the prognosis of HBV-ACLF after ALSS therapy, i.e., TBRR-ICGR 15 , TBARR-ICGR 15 , TBARR-ICGR 15 -ΔMELD, and TBARR-ICGR 15 -ΔMELD-age, with an AUC of 0.830, 0.867, 0.900, and 0.917, respectively, and the combined predictive models had a larger AUC than each index alone (age, neutrophil count, TBRR, TBARR, ΔMELD, MELD score, CTP score, and ICGR 15 ), among which the TBARR-ICGR 15 -ΔMELD-age model had the largest AUC. The combined models TBARR-ICGR 15 -ΔMELD and TBARR-ICGR 15 -ΔMELD-age had sensitivities and specificities of > 80%. Conclusion The combined predictive model established by ICGR 15 and TBARR has a good value for in predicting the short-term prognosis of patients with HBV-ACLF after ALSS therapy, and the combined predictive model has a better accuracy than the single model in judging prognosis.