Objective To investigate the correlation between liver and spleen stiffness measured by acoustic radiation force impulse (ARFI) and hepatic venous pressure gradient (HVPG),and to evaluate its efficiency in the diagnosis of portal hypertension.Methods From April 2014 to March 2016,20 cases underwent HVPG measurement because of liver cirrhosis were enrolled.Before HVPG measurement,liver and spleen stiffness were assessed with ARFI.The correlation between HVPG and age,alanine aminotransferase (ALT),aspartate aminotransferase (AST),total hilirubin,serum albumin,platelet count,prothrombin time,aspartate aminotransferase to platelet ratio index (APRI) score,Child-Pugh score,model for end-stage liver disease (MELD) score,liver stiffness and spleen stiffness were analyzed.Pearson correlation and Spearman rank correlation were performed for statistical analysis.Results HVPG,liver and spleen stiffness were successfully measured in all 20 patients.The mean liver stiffness was (1.78±0.29) m/s,the mean spleen stiffness was (3.37±0.44) m/s and HVPG was (16.10±5.14) mmHg (1 mmHg=0.133 kPa).Age,ALT,AST,total bilirubin,serum albumin,platelet count,prothrombin time,APRI score,Child-Pugh score and MELD score were all not correlated with HVPG (all P＞0.05).But HVPG was positively correlated with liver and spleen stiffness (r=0.449,P=0.047;r=0.487,P=0.030).In the diagnosis of HVPG≥12 mmHg,the area under curve (AUC) of liver stiffness was 0.875,the optimal cut-off value was 1.77 m/s,the sensitivity was 68.6 ％ and the specificity was 100.0％.In the diagnosis of HPVG≥20 mmHg,the AUC of liver stiffness was 0.798,the optimal cut off value was 1.85 m/s,the sensitivity was 100.0％ and the specificity was 68.8％.The AUC of spleen stiffness was 0.820,the optimal cut-off value was 3.23 m/s,the sensitivity was 100.0 ％ and the specificity was 56.3％.Conclusion In patients with liver cirrhosis,liver stiffness and spleen stiffness assessed by ARFI are positively correlated with HVPG and therefore ARFI has certain application value in the noninvasive diagnosis of portal hypertension.

Objective To discuss the influence of microRNA(miR)-155/miR-21 on toll-like receptor 4 (TLR4) in children with sepsis.Methods Fifty children with sepsis who were hospita-lized in Pediatric Intensive Care Unit,Shenzhen Children's Hospital,were enrolled in the study,and 15 healthy children at the same age were selected as healthy control group.Expression levels of TLR4 protein and human leukocyte antigen(HLA)-DR in CD14 + monocytes (MC) were detected by using flow cytometry,and sepsis patients were divided into 2 groups according to whether they exceeded the value of HLA-DR by 30％ or not.Expression level of programmed cell death factor 4 (PDCDM) and inositol phosphatases 1 containing SH2 (SHIP1) were detected at the same time.MC were separated by CD14 + immune magnetic bead,and expression level of miR-155,miR-21 and tumor necrosis factor-α (TNF-α),interleukin-10 (IL-10) mRNA in CD14 + MC were detected by using real-time fluorescent quantitative PCR.Results Sepsis group consisted of 27 male and 23 female,and their ages were (2.34 ± 0.79) years old,among whom 9 patients died.There were 36 patients in the HLA-DR increase group and 14 patients in the HLA-DR decrease group.Expressions ofTLR4(2.33±0.90),miR-155[(7.19±3.75) ×10 3] and TNF-α[(21.98±14.15) ×10-2 pg/L] in CD14 + MC were higher in the HLA-DR increase group than those in the HLA-DR decrease group [1.24±0.60,(4.83 ±1.17) × 10-3,(14.18±5.45) ×10-2 μg/L] and healthy control group[1.57±0.55,(3.99 ± 1.29) × 10-3,(1.61 ± 0.84) × 10 2 pg/L],and the differences were statistically significant(F =11.943,7.583,18.538,all P ＜0.05),while the expressions of miR-21 (12.10 ±5.66),IL-10[(29.74 ± 12.55) × 10-4 μg/L] in CD14 + MC were lower in the HLA-DR increase group than those in the HLA-DR decrease group[4.68 ± 2.07,(12.50 ± 5.73) × 10-4 μg/L] and healthy control group [2.39 ± 0.86,(2.04 ± 0.92) × 10-4 μg/L],and the differences were statistically significant(F =41.673,54.991,all P ＜ 0.05).The levels of SH1P1 and PDCD4 decreased in sepsis compared with healthy control group[0.70 ±0.36)vs.(1.59 ±0.48);(1.55 ±0.56) vs.(3.01 ±0.70)],and the differences were statistically significant (t =7.682,8.339,all P ＜ 0.05),but SHIP1 decreased more significantly in the HLA-DR increase group than that in the HLA-DR decrease group [(0.60 ± 0.34) vs.(0.97 ± 0.26)],and the difference was statistically significant (F =39.214,P ＜ 0.05).PDCD4 decreased more significantly in the HLA-DR decrease group than that in the HLA-DR increase group (0.94 ±0.19 vs.1.79 ±0.47),the difference was statistically significant(F =65.367,P ＜ 0.05).Conclusions Regulation imbalance of miR-155/miR-21 may be one of the reasons for abnormal expression of TLR4 in children with sepsis,and it plays a role in enlarged or inhibited expression of TLR4 in the sepsis process which results in different immune status in sepsis patients.

Objective To determine the mortality and associated risk factors in the patients with diabetic foot ulcers. Methods One hundred and sixty-three patients with diabetic foot ulcers hospitalized from January 2001 to December 2006 were followed up until December 2009. Mortality rates were derived from Kaplan-Meier survival curves. The prognostic factors were evaluated with Cox proportional hazard model. Results Follow-up was successful in 139 out of 163 patients, with a mean follow-up period of(3.71 + 1. 80)years. 55 patients(39 males and 16 females)died during the follow-up. The 5-year mortality was 45.8% and mean survival time was 5.38 years(95% CI 4.87-5.89). The median survival time was 6.83 years. Age, smoking, hypertension, coronary artery disease, and diabetic nephropathy were found to be independent prognostic factors for mortality. Conclusions Diabetic foot ulcers increased the mortality of diabetic patients. Age, smoking, hypertension, coronary artery disease, and diabetic nephropathy were predictive risk factors for mortality.

Aim To study targeting capability of anti-CD19 (Fab)-LDMto CD19 +B lymphoma cells in vi-vo and in vitro.Methods Flow cytometry was em-ployed to determine the affinity of Cy5 labeled anti-CD19 (Fab)-LDP to human lymphoma Raji cells.And the optical imaging system was used to analyze the dis-tribution of Cy5-anti-CD19 (Fab )-LDP in lymphoma-transplanted xenograft nude mice in vivo.Results The results of flow cytometry demonstrated that Cy5-an-ti-CD19(Fab)-LDP had remarkable affinity with lym-phoma Raji cells;Raji lymphoma xenograft model was established successfully in nude mice and in vivo fluo-rescence imaging analysis indicated that the antibody-drug conjugates could specially be localized in the tar-get tumor.Conclusion The experiments in vivo and vitro confirm that anti-CD19 (Fab)-LDP has remarka-ble affinity to targeting CD19 +lymphoma cells,and the antibody drugs anti-CD19 (Fab )-LDP have the probability to be new drugs for the treatment of malig-nant lymphoma.

Objective To study the clinical characteristics of gynura segetum induced hepatic sinusoidal obstruction syndrome (HSOS).Methods From July 2008 to October 2016,a total of 115 cases of gynura segetum caused HSOS were retrospectively analyzed.The history of taking gynura segetum before disease onset was recorded and epidemiologic data of main clinical symptoms,clinical manifestations,laboratory examinations,imaging and pathological features were observed.Results Among the 115 cases of HSOS,there were 113 patients with abdominal pain,106 with anorexia and 42 with jaundice sclera.A total of 108 patients displayed increased serum total bilirubin,41 of them only with mildly increased total bilirubin.There were 29 patients with albumin lower than 30 g/L,64 patients with prolonged prothrombin time (PT) and PT of 11 patients was prolonged for more than three seconds.Meanwhile,31 patients were with prolonged activated partial prothrombin time (APTT).A total of 60 patients had low platelet count.And 92 patients underwent ultrasound examination,among them,71 patients had enlarged liver size,79 patients with uneven internal echo of liver,70 patients with ascites,14 patients with patchy low echo tissue around hepatic venous.A total of 60 patient accepted computed tomography (CT) examination,and all of them had ascites,14 patients with mildly enlarged spleen and eight patients with gastro-esophageal varices.The results of CT plain scan indicated hepatomegaly,decreased liver density,map-like changes of patchy low density in delayed phase,heterogeneous enhancement of liver parenchyma in arterial phase,compression and deformation of liver segment of inferior vena cava and halo sign around venous portal vein.The results of pathological examination demonstrated the widening of hepatic sinusoid with hemorrhage and congestion,destruction of liver plate in zone Ⅲ area.There were seven patients who received hepatic venous pressure gradient (HVPG) measurement which were all significantly increased.Conclusions The characteristics of patients with gynura segetum caused HSOS are abdominal pain,anorexia and jaundice;mildly increased serum total bilirubin and albumim liver enlargement,slow blood velocity of portal vein and splenic veim increased HVPG,hepatic sinus congestion and cell coagulation necrosis in zone Ⅲ area.

Objective:To explore the role of serum pyrrole-protein-adduct (PPA) in evaluating the severity and predicting the anticoagulant efficacy in patients with pyrrolidine alkaloid-related hepatic sinusoidal obstruction syndrome (PA-HSOS).Methods:From April 2018 to December 2019, the data of 48 patients with PA-HSOS admitted and treated at Drum Tower Hospital, Affiliated Medical College of Nangjing University were collected, which included PPA level, portal vein velocity (PVV), ascites grading, PA-HSOS severity grading (according to the new severity grading criteria for suspected hepatic sinusoidal obstruction syndrome in adults by the European Society of Blood and Bone Marrow Transplantation and adjusted) and the outcome of anticoagulation. Patients with acute onset (onset of symptoms within 1 month after consuming pyrrolizidine alkaloid-containing plants) were taken as research subjects. The combination of PPA with PVV or with ascites classification of PA-HSOS severity assessment model was fitted by logistic regression, and the logit values of 2 combination models were calculated, the formula was logit 1=0.034×PPA(nmol/L)+ 0.055×PVV(cm/s)-3.287, logit 2=0.039×PPA(nmol/L)-2.712×ascites grade 2 (Yes=1, No=0)-0.388×ascites grade 3 (Yes=1, No=0)-0.899. The patients received initial anticoagulation therapy at Drum Tower Hospital, Affiliated Medical College of Nanjing University were selected as research subjects. The anticoagulant efficacy prediction model of combination of PPA with serum creatinine (SCR) and with hepatic venous pressure gradient (HVPG) was fitted by logistic regression, and the logit value was calculated, the formula was logit 3=0.013×PPA(nmol/L)+ 0.064×SCR (mol/L)+ 0.542×HVPG (mmHg, 1 mmHg=0.133 kPa)-16.005. The predictive value of PPA in evaluating the severity of PA-HSOS and anticoagulant efficacy was evaluated. Receiver operating characteristic curve analysis was performed for statistical analysis. Results:The serum PPA level of 48 patients was 10.81 nmol/L (3.91 nmol/L, 32.04 nmol/L). Among them, 33 cases (68.8%) were mild PA-HSOS, 3 cases (6.2%) were moderate PA-HSOS, no severe PA-HSOS case and 12 cases (25.0%) were very severe PA-HSOS. Among 23 patients received initial anticoagulant therapy at Drum Tower Hospital, Affiliated Medical College of Nanjing University and with complete data, 8 patients responded and survived, and 15 patients did not respond (5 patients died, 1 patient relieved after continue anticoagulant therapy, and 9 patients survived after switching to anticoagulant therapy and transjugular intrahepatic portosystemic shunt (TIPS) treatment). One patient without initial anticoagulant therapy, survived after TIPS treatment because of the progress of the disease. Area under the curve (AUC) of PPA to assess the severity of acute onset PA-HSOS was 0.75, 95% confidence interval ( CI) was 0.52 to 0.98 ( P=0.047). When PPA≥45.519 nmol/L, the specificity and sensitivity in evaluating severe and very severe PA-HSOS was 100.0% and 57.1%, respectively. AUC of combination of PPA and PVV to assess the severity of PA-HSOS was 0.77, 95% CI was 0.55 to 1.00 ( P=0.032). When the logit of combination model≥0.180, the specificity and sensitivity in evaluating severe and very severe PA-HSOS was 71.4% and 81.8%, respectively. AUC of combination of PPA and ascites grade (grade 1, 2 or 3) to assess the severity of PA-HSOS was 0.85, 95% CI was 0.63 to 1.00 ( P=0.005). When the logit of combination model≥0.347, the specificity and sensitivity in evaluating severe and very severe PA-HSOS was 85.7% and 92.0%, respectively. AUC of combination of PPA, SCR and HVPG to predict anticoagulation efficacy was 0.85, 95% CI was 0.69 to 1.00 ( P=0.009). When the logit≥0.393, the specificity and sensitivity in predicting anticoagulation efficacy was 62.5% and 91.7%, respectively. Conclusions:PPA can be used to assess the severity of acute onset PA-HSOS patients, and combined with ascites grading can significantly improve its efficiency. PPA combined with SCR and HVPG can better predict anticoagulant efficacy.

Objective To explore the causal relationship between rheumatoid arthritis(RA)and iron deficiency a-nemia(IDA)in European population by two-sample Mendelian randomization analysis.Methods The single nu-cleotide polymorphisms(SNPs)of RA and IDA were analyzed using public genome-wide association studies(GWAS).The inverse variance weighting method(IVW)was used as the main analysis method to evaluate the causal effect of RA on IDA.MR-Egger method,weighted median method(WM),weighted model method and simple model method were used as regression supplements to evaluate the robustness of sensitivity analysis results.The het-erogeneity function was used to calculate the P-value to test the heterogeneity,and the intercept term intercept was used to test the level pleiotropy.Results In the FINNGEN database at the genome-wide level,strong-related SNPs that removed linkage disequilibrium and met the P＜5.0 × 10-8 by Mendelian randomization analysis were select-ed.After integrating exposure and outcome data,31 SNPs were obtained as the final effective instrumental variables.IVW showed that RA was a risk factor for IDA(the risk of IDA in RA patients was 1.064 times higher than that in non-RA patients,OR=1.064,95％CI:1.028-1.103).The weighted median method and MR-Egger method re-sults supported the positive correlation between RA and IDA.The intercept value was close to 0,indicating that there was no horizontal pleiotropy between exposure and outcome.The heterogeneity function's P＜0.05 indicated that there was heterogeneity between exposure and outcome,but the random effect model test showed P＜0.05,indi-cating that even if there was heterogeneity in causality,the overall trend was stable.Conclusion RA is a risk factor for IDA,and there is a positive correlation between RA and IDA.

The brain has very high energy requirements and consumes 20% of the oxygen and 25% of the glucose in the human body. Therefore, the molecular mechanism underlying how the brain metabolizes substances to support neural activity is a fundamental issue for neuroscience studies. A well-known model in the brain, the astrocyte-neuron lactate shuttle, postulates that glucose uptake and glycolytic activity are enhanced in astrocytes upon neuronal activation and that astrocytes transport lactate into neurons to fulfill their energy requirements. Current evidence for this hypothesis has yet to reach a clear consensus, and new concepts beyond the shuttle hypothesis are emerging. The discrepancy is largely attributed to the lack of a critical method for real-time monitoring of metabolic dynamics at cellular resolution. Recent advances in fluorescent protein-based sensors allow the generation of a sensitive, specific, real-time readout of subcellular metabolites and fill the current technological gap. Here, we summarize the development of genetically encoded metabolite sensors and their applications in assessing cell metabolism in living cells and in vivo, and we believe that these tools will help to address the issue of elucidating neural energy metabolism.
Animals ; Biosensing Techniques ; Brain ; cytology ; metabolism ; Cytological Techniques ; Energy Metabolism ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Time Factors

Objective To investigate the accuracy of liver stiffness (LS) as a noninvasive index in predicting hepatic venous pressure gradient (HVPG) in patients with decompensated liver cirrhosis and the value of LS in the diagnosis of decompensated liver cirrhosis. Methods A retrospective analysis was performed for the clinical data of 88 patients with decompensated cirrhosis due to viral hepatitis or decompensated alcoholic cirrhosis who received both HVPG measurement and LS measurement by acoustic radiation force impulse (ARFI) in Department of Gastroenterology, Nanjing Drum Tower Hospital, from April 2013 to June 2021, and according to HVPG, the patients were divided into serious portal hypertension (SPH) (HVPG≥20 mmHg) group with 24 patients and non-SPH (HVPG < 20 mmHg) group with 64 patients. The two groups were compared in terms of LS, spleen stiffness, portal vein velocity, and related biochemical parameters. The t -test or the Mann-Whitney U rank sum test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. A Pearson correlation analysis was used to investigate the correlation of different noninvasive indices with HVPG, and a Logistic regression analysis was used to investigate the association of different noninvasive indices with the risk of SPH. Receiver operating characteristic (ROC) curves were plotted for different noninvasive indices in predicting HVPG≥20 mmHg, and the area under the ROC curve (AUC), sensitivity, specificity, maximum Youden index, and corresponding cut-off value were calculated to investigate the value of each index in predicting SPH. Results Among the 88 patients, 76 had decompensated cirrhosis due to viral hepatitis and 12 had decompensated alcoholic cirrhosis. There were no significant differences between the SPH group and the non-SPH group in age, sex, white blood cell count, hemoglobin, platelet count, prothrombin time, alanine aminotransferase, aspartate aminotransferase, albumin, serum sodium, creatinine, Child-Pugh class, and spleen stiffness, while there was a significant difference in LS between the two groups ( t =-3.970, P < 0.01). The correlation analysis showed that HVPG was positively correlated with LS ( r =0.458, P < 0.001). The Logistic regression analysis showed that LS was a risk factor for SPH (odds ratio=3.941, 95% confidence interval: 1.245-12.476, P =0.020). The ROC curve analysis showed that LS had an AUC of 0.751 in predicting the onset of SPH, with a sensitivity of 54.17% and a specificity of 90.63% at the optimal cut-off value of 2.295 m/s. Conclusion In patients with decompensated cirrhosis, LS measured by ARFI is correlated with HVPG and has a certain value in the non-invasive diagnosis of decompensated cirrhosis with HVPG≥20 mmHg.