Objective:To explore the effectiveness and safety of different anticoagulation strategies during continuous blood purification (CBP) treatment,providing a reference for anticoagulation strategies in critically ill children undergoing CBP.Methods:A retrospective study was conducted,including children admitted to the PICU of Children's Hospital of Fudan University from January 2019 to December 2024.According to the anticoagulation methods used during CBP treatment,patients were divided into the sodium citrate group and the heparin group.CBP was performed using continuous venovenous hemofiltration or continuous venovenous hemodialysis filtration mode,with a blood flow rate of 3-5 mL/(kg·min),replacement fluid rate of 30-50 mL/(kg·h),and dialysis fluid rate of 20-30 mL/(kg·h).The filter lifespan,28-day all-cause mortality,total length of hospital stay,PICU stay duration,adverse events,and associated costs were compared between the two groups.Results:A total of 221 children were included (105 in the sodium citrate group and 116 in the heparin group),with a cumulative use of 666 filters (284 in the sodium citrate group and 382 in the heparin group).(1) There were no statistically significant differences in general data,including age,sex ratio,underlying diseases,the ratio and duration of invasive mechanical ventilation,vasopressor scores at baseline,and indications for CBP between the two groups (all P>0.05).(2) The filter lifespan was 20(14,32) hours for the sodium citrate group and 21(13,35) hours for the heparin group,with no statistically significant difference between the two groups ( P>0.05); the proportion of accidental downstroke was 2.8% and 6.5%,respectively,with a statistically significant difference ( P=0.029); among the 221 children,86 died,with 38 deaths (35.2%) in the sodium citrate group and 49 deaths (38.9%) in the heparin group,showing no statistically significant difference.(3) The sodium citrate group had a higher incidence of metabolic alkalosis,hypocalcemia,and sodium citrate accumulation (44.4% vs. 1.6%,32.7% vs 9.4%,7.7% vs 0,all P<0.01); the heparin group had a greater proportion of bleeding (6.0% vs. 2.9%) and was more likely to develop heparin-induced thrombocytopenia (10.2% vs. 0, P<0.01).(4) The total hospitalization costs for the sodium citrate group were significantly higher than for the heparin group (200 327 yuan vs. 152 077 yuan, P=0.05); costs related to the use of anticoagulants and monitoring indicators during CBP treatment were also higher in the sodium citrate group (2 479 yuan vs. 682 yuan, P<0.01). Conclusions:Sodium citrate is a safe and effective anticoagulation method for critically ill children undergoing CBP,which can reduce the risk of filter clotting compared to systemic heparin anticoagulation.

Objective:To explore the effectiveness and safety of different anticoagulation strategies during continuous blood purification (CBP) treatment,providing a reference for anticoagulation strategies in critically ill children undergoing CBP.Methods:A retrospective study was conducted,including children admitted to the PICU of Children's Hospital of Fudan University from January 2019 to December 2024.According to the anticoagulation methods used during CBP treatment,patients were divided into the sodium citrate group and the heparin group.CBP was performed using continuous venovenous hemofiltration or continuous venovenous hemodialysis filtration mode,with a blood flow rate of 3-5 mL/(kg·min),replacement fluid rate of 30-50 mL/(kg·h),and dialysis fluid rate of 20-30 mL/(kg·h).The filter lifespan,28-day all-cause mortality,total length of hospital stay,PICU stay duration,adverse events,and associated costs were compared between the two groups.Results:A total of 221 children were included (105 in the sodium citrate group and 116 in the heparin group),with a cumulative use of 666 filters (284 in the sodium citrate group and 382 in the heparin group).(1) There were no statistically significant differences in general data,including age,sex ratio,underlying diseases,the ratio and duration of invasive mechanical ventilation,vasopressor scores at baseline,and indications for CBP between the two groups (all P>0.05).(2) The filter lifespan was 20(14,32) hours for the sodium citrate group and 21(13,35) hours for the heparin group,with no statistically significant difference between the two groups ( P>0.05); the proportion of accidental downstroke was 2.8% and 6.5%,respectively,with a statistically significant difference ( P=0.029); among the 221 children,86 died,with 38 deaths (35.2%) in the sodium citrate group and 49 deaths (38.9%) in the heparin group,showing no statistically significant difference.(3) The sodium citrate group had a higher incidence of metabolic alkalosis,hypocalcemia,and sodium citrate accumulation (44.4% vs. 1.6%,32.7% vs 9.4%,7.7% vs 0,all P<0.01); the heparin group had a greater proportion of bleeding (6.0% vs. 2.9%) and was more likely to develop heparin-induced thrombocytopenia (10.2% vs. 0, P<0.01).(4) The total hospitalization costs for the sodium citrate group were significantly higher than for the heparin group (200 327 yuan vs. 152 077 yuan, P=0.05); costs related to the use of anticoagulants and monitoring indicators during CBP treatment were also higher in the sodium citrate group (2 479 yuan vs. 682 yuan, P<0.01). Conclusions:Sodium citrate is a safe and effective anticoagulation method for critically ill children undergoing CBP,which can reduce the risk of filter clotting compared to systemic heparin anticoagulation.

Background and Objective Accurate diagnosis of mucous cystic lesion(PCL)remains a clinical difficult.Both Carcinoembryonic antigen(CEA)and glucose(GLU)are reported to have ability to distinguish mucinous PCL from non-mucinous PCL,but the accuracy was limited.The objective of this study was to evaluate the value of cystic CEA combined with GLU in the diagnosis of mucinous PCL.Methods PCL patients who underwent pancreatic surgery and endoscopic ultrasonography guided fine-needle aspiration(EUS-FNA)were retrospectively collected from the First Affiliated Hospital of Naval Medical University.Clinical data and cystic fluid analysis of included PCLs patients were analyzed using receiver operator(ROC)curves.ROC analysis,sensitivity and specificity analyses were used to evaluate the value of CEA combined with GLU in the diagnosis of mucinous PCL.Results From January 2015 to December 2021,a total of 84 patients underwent cyst fluid CEA and GLU analysis,of whom 44(52.4％)had mucinous PCL and 40(47.6％)had non-mucinous PCL.The AUC for distinguishing mucinous from non-mucinous PCL by CEA was 0.82[(95％confidence interval(CI):0.72-0.92)].When 192 ng/mL was used as the cutoff level,the diagnostic sensitivity and specificity were 50％and 93％,respectively.Using 20 ng/mL as cutoff level,the diagnostic sensitivity increased to 80％and the specificity decreased to 68％.The AUC for the cystic GLU to distinguish mucinous from non-mucinous PCL was 0.73(95％CI:0.99-0.87),and the diagnostic sensitivity and specificity were 100％and 60％,respectively.When the cutoff level of CEA was 192 ng/mL,the AUC of CEA combined with GLU in the diagnosis of mucinous PCLs was 0.94(95％Cl:0.86-0.99),while when the cutoff level of CEA was 20 ng/mL,the AUC of CEA combined with Glu in the diagnosis of mucinous PCLs was 0.94(95％CI:0.85-0.99).The AUCs were significantly higher than the AUC with single diagnostic indicators.Conclusion When using the cutoff level of 192 ng/mL,cyst fluid CEA combined with GLU has high sensitivity and specificity in differentiating mucinous PCL from non-mucinous PCL,so it can be considered for clinical application.Lower CEA cutoff level(20 ng/mL)can improve the sensitivity of diagnosis.

Objective:To compare the characteristics of patients undergoing blood purification treatment in PICU of a children′s tertiary hospital during 8 years, so as to analyze the changes in the development of blood purification technology in children in East China.Methods:Patients who received blood purification treatment in PICU of Children′s Hospital of Fudan University from 2014 to 2021 were included and divided into two study periods: 2014-2017 and 2018-2021.The clinical characteristics and treatment parameters of patients were collected and analyzed.Results:A total of 1 029 patients were included in the study, of which 103 were combined with extracorporeal membrane oxygenation.The 28-day survival rate of 926 patients treated with pure blood purification was 55.7%.Among them, patients with younger age, lower body weight, using mechanical ventilation, using vasoactive drugs before blood purification, and patients with multiple organ dysfunction syndrome had a higher distribution in the death group than those in survival group( P<0.05). During 8 years, a total of 3 688 cases of blood purification were performed.The main mode was continuous veno-venous hemodiafiltration (CVVHDF) (68.6%), followed by therapeutic plasma exchange (TPE) (23.8%) and hemoperfusion (HP) (4.8%); the main indication was acute kidney injury (AKI) (29.3%), followed by severe inflammatory disease (26.2%) and acute liver failure (16.2%). Compared with 2014-2017, the number of blood purification treatments in 2018-2021 increased by 47.4%, and the survival rate of patients increased significantly (48.7% vs. 58.1%, P<0.05). The distribution of blood purification patterns and indications also changed( P<0.05). The proportions of TPE (20.5% vs. 26.0%) and HP (3.1% vs. 6.0%) increased, while the proportion of CVVHDF (71.9% vs. 66.4%) decreased significantly.The proportions of AKI (29.8% vs. 38.9%) and refractory immune diseases (8.4% vs. 15.2%) were significantly higher, while severe inflammatory diseases (29.2% vs. 24.2%) and acute liver failure (19.6% vs. 13.8%) had declined. Conclusion:From 2014 to 2021, the number of blood purifications performed in our center increased significantly.Although the distribution of indications and patterns have also changed significantly, the overall survival rate is significantly improved.However, standardized practice still needs to be strengthened.

Objective:To explore whether the lipopolysaccharide (LPS)-induced modification of O-linked N-acetylglucosamine (O-GlcNAc) is involved in the inflammatory signaling pathway of endothelial cells.Methods:Human umbilical vein endothelial cells (HUVEC) were cultured in vitro, and cells in logarithmic growth phase were used for experiments. Cells were divided into blank control group, LPS group (2 000 mg/L LPS), O-GlcNAc transferase (OGT) overexpression (OGT-OE)+LPS group (plasmid transfection OGT+2 000 mg/L LPS), protein kinase C (PKC) inhibitor+LPS group (10 μmol/L Go 6983+2 000 mg/L LPS), RhoA inhibitor+LPS group (40 μmol/L Rhoin hydrochloride+2 000 mg/L LPS), phosphatidylinositol-3-kinase (PI3K) inhibitor+LPS group (1 μmol/L SL-2052+2 000 mg/L LPS), serine/threonine kinase (Akt) inhibitor+LPS group (10 μmol/L PP2+2 000 mg/L LPS) and small interfering RNA (siRNA) treated Akt (si-AKT)+LPS group (si-Akt+2 000 mg/L LPS). After 24 hours of LPS treatment, real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the transcription levels of inflammatory cytokines [interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)]. The protein expression or phosphorylation of OGT, O-GlcNAc, Akt, extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein kinase (p38MAPK), nuclear factor-κB p65 (NF-κB p65), and signal transducer and activator of transcription 3 (STAT3) were determined by Western blotting. Results:Compared with the blank control group, the expression of OGT and the modification of O-GlcNAc in the LPS group were decreased, while the expressions of phosphorylated ERK, p38MAPK, and STAT3 were increased, and the transcript levels of inflammatory cytokines were also significantly increased [IL-6 mRNA (2 -ΔΔCt): 4.71±0.60 vs. 1.03±0.29, TNF-α mRNA (2 -ΔΔCt): 1.89±0.11 vs. 1.04±0.35, ICAM-1 mRNA (2 -ΔΔCt): 2.06±0.18 vs. 1.02±0.21, VCAM-1 mRNA (2 -ΔΔCt): 2.94±0.57 vs. 1.01±0.17, all P < 0.05], indicating that LPS could decrease O-GlcNAc modification, activate inflammatory signaling pathways and increase inflammatory cytokines expression. Compared with the LPS group, the expressions of phosphorylated ERK, p38MAPK, NF-κB p65, and STAT3 in the endothelial cells of the OGT-OE+LPS group were decreased, and the expression of inflammatory factors were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.12±0.01 vs. 0.90±0.17, TNF-α mRNA (2 -ΔΔCt): 0.31±0.01 vs. 0.91±0.14, ICAM-1 mRNA (2 -ΔΔCt): 0.64±0.02 vs. 1.13±0.16, VCAM-1 mRNA (2 -ΔΔCt): 0.11±0.01 vs. 0.93±0.11, all P < 0.05], indicating that the increase of OGT level could inhibit the partial activation of the endothelial inflammatory signal pathway under the LPS stimulation. Compared with the blank control group, the phosphorylation level of Akt in the LPS group was increased. Compared with the LPS group, both OGT expression and O-GlcNAc modification were down-regulated after pretreatment of PKC inhibitor, RhoA inhibitor, PI3K inhibitor, or Akt inhibitor. Compared with the LPS group, the transcript levels of IL-6, TNF-α and ICAM-1 in the PP2+LPS group were significantly decreased [IL-6 mRNA (2 -ΔΔCt): 1.46±0.16 vs. 3.55±0.87, TNF-α mRNA (2 -ΔΔCt): 0.98±0.14 vs. 1.76±0.10, ICAM-1 mRNA (2 -ΔΔCt): 1.39±0.24 vs. 2.04±0.13, all P < 0.05], but there was no significant change in VCAM-1. Compared with the LPS group, the expression of OGT and O-GlcNAc modification in the si-Akt+LPS group were decreased, while the transcript levels of inflammatory cytokines were also significantly decreased [IL-6 mRNA (2 -ΔΔCt): 0.75±0.03 vs. 0.99±0.09, TNF-α mRNA (2 -ΔΔCt): 0.69±0.01 vs. 1.10±0.08, ICAM-1 mRNA (2 -ΔΔCt): 0.76±0.01 vs. 0.99±0.02, VCAM-1 mRNA (2 -ΔΔCt): 0.93±0.08 vs. 1.20±0.21, all P < 0.05], indicating that Akt participated in the action process of LPS on OGT and affected the inflammatory factor expression. Conclusions:The decreased level of O-GlcNAc modification in endothelial cells stimulated with LPS promotes partial activation of inflammatory signaling pathways, mainly involving ERK, p38MAPK, and STAT3, and affects the expression of inflammatory factors. AKT may be involved in the effect of LPS on the inhibition of O-GlcNAc modification.

Trifluridine-tipiracil(TAS-102)is an oral chemotherapeutic agent combination comprising a fluoropyrimidine(FTD)and a potent thymidine phosphorylase(TPI)inhibitor.TAS-102 is novel due to its antitumor activity against 5-fluorouracil resistant tumors,demonstrated both by in vitro models and xenografts.TAS-102 is currently approved as a third-line therapeutic strategy for metastatic colorectal and gastric cancer based on phase Ⅲrandomized clinical trial data confirming survival benefits with TAS-102.Preliminary data from recent studies suggest a potential expanding role of TAS-102 in various gastrointestinal(GI)cancers.This article summarizes the clinical applications of TAS-102 in the treatment of GI cancers.Additionally,we discuss the rationale and clinical benefits of combining TAS-102 with other anticancer agents in patients with GI cancers.

The regulator of expression of virion(Rev)protein binds specifically to the Rev-responsive element(RRE)RNA in order to regulate the expression of the human immunodeficiency virus(HIV)-1 genes.Fluores-cence indicator displacement assays have been used to identify ligands that can inhibit the Rev-RRE interaction;however,the small fluorescence indicators cannot fully replace the Rev peptide or protein.As a result,a single rhodamine B labeled Rev(RB-Rev)model peptide was utilized in this study to develop a direct and efficient Rev-RRE inhibitor screening model.Due to photon-induced electron transfer quenching of the tryptophan residue on the RB fluorophore,the fluorescence of RB in Rev was weakened and could be dramatically reactivated by interaction with RRE RNA in ammonium acetate buffer(approximately six times).The interaction could reduce the electron transfer between tryptophan and RB,and RRE could also increase RB fluorescence.The inhibitor screening model was evaluated using three known positive Rev-RRE inhibitors,namely,proflavin,6-chloro-9-[3-(2-chloroethylamino)pro-pylamino]-2-methoxyacridine(ICR 191),and neomycin,as well as a negative drug,arginine.With the addition of the positive drugs,the fluorescence of the Rev-RRE decreased,indicating the displacement of RB-Rev.This was confirmed using atomic force microscopy(AFM)and the fluorescence was essentially unaffected by the addition of arginine.The results demonstrated that RB-Rev can be used as a fluorescent probe for recognizing small ligands that target RRE RNA.The Rev-RRE inhibitor screening model offers a novel approach to evaluating and identifying long-acting Rev inhibitors.

Metastatic colorectal cancer (mCRC) is a clinical and molecular heterogeneous disease. Currently, for mCRC, extended rat sarcoma (RAS) testing is recommended in routine clinical practice before any treatment. RAS mutational status is significantly associated with the outcome of patients and strongly predictive for anti-EGFR-targeted therapy. However, specific treatments for RAS target are not yet available. Previous studies have shown that direct inhibition of RAS proteins has limited clinical benefits. Recently, a promising drug, AMG-510, which can directly inhibit KRAS G12C has been reported; however, it needs further confirmation. In the past few years, important advances have also been made in approaches designed to indirectly target RAS by inhibiting RAS effectors, multi-target combination strategies and immunotherapy. They are expected to be effective treatments for RAS target. This article summarizes the precision treatment of RAS-mutant mCRC.

Adenovirus pneumonia is a common type of pneumonia in immunocompetent and immunocompromised patients, and its prognosis is poor.Antiviral therapy includs ganciclovir, valganciclovir, cidofovir, brincidofovir, ribavirin and so on.Among them, cidofovir and brincidofovir have obvious antiviral activity against adenovirus in vitro and in vivo, but further RCT results are still needed.Therefore, antiviral therapy of adenovirus pneumonia still needs further study.

The mortality of sepsis remains high,and it is still a major cause of death in critically ill patients.But its pathophysiological mechanism remains a mystery,not yet fully clear.Endothelium play an important role in the development of sepsis.Maintaining the integrity of its surface glycocalyx to a certain extent reduces sepsis damage;and its destruction mediates a series of pathophysiological responses,including increasing vascular permeability by cell pathway or intracellular pathways,increasing the adhesion of white blood cells to endothelial cejls and a variety of cytokines to promote inflammatory response,activating of thrombin to coagulation dysfunction,promoting synthesis of nitric oxide to change vascular tension.The damage of glycocalyx as a first step in endothelial cell injury can be used as early diagnosis and prognosis of sepsis,which provides new ideas for the diagnosis and treatment of sepsis.