Molecular Detection of FGFR2 Rearrangements in Resected Intrahepatic Cholangiocarcinomas:FISH Could Be An Ideal Method in Patients with Histological Small Duct Subtype

Background and Aims:Intrahepatic cholangiocarcinoma(ICC)is a subtype of primary liver cancer for which effective therapeutic agents are lacking.Fibroblast growth factor receptor 2(FGFR2)has become a promising therapeutic target in ICC;however,its incidence and optimum testing method have not been fully assessed.This study investigated the rearrangement of FGFR2 in intrahepatic cholangiocarcinoma using multiple molecular detection methods.Methods:The samples and clinical data of 167 patients who underwent surgical resection of intrahepatic cholangiocarcinoma in Zhongshan hospital,Fudan university were collected.The presence of FGFR2 gene rearrangement was confirmed using fluorescence in situ hybridization(FISH)and targeted next-generation sequencing(NGS).FGFR2 protein expression was determined using immunohistochemistry(IHC).The concordance between the methods was statistically compared.PD-L1 expression was also assessed in this cohort.The clinicopathological characteristics and genomic profile related to FGFR2 rearrangements were also analyzed to assist candidatescreening for targeted therapies.Results:FGFR2 rearrangement was detected in 21 of the 167 ICC cases(12.5%)using FISH.NGS analysis revealed that FGFR2 rearrangement was present in 16 of the 20 FISH-positive cases,which was consistent with the FISH results(kappa value=0.696,p<0.01).IHC showed that 80 of the 167 cases(48%)were positive for FGFR2 expression,which was discordant with both FISH and NGS results.By comparison,FGFR2-positivity tended to correlate with unique clinicopathological subgroups,featuring early clinical stage,histologically small duct subtype,and reduced mucus production(P<0.05),with improved overall survival(p<0.05).FGFR2-positivity was not associated with PD-L1 expression in ICCs.In genome research,we identified eight partner genes fused with FGFR2,among which FGFR2-BICC1 was the most common fusion type.BAP1,CDKN2A,and CDKN2B were the most common concomitant genetic alterations of FGFR2,whereas KRAS and IDH1 mutations were mutually exclusive to FGFR2 rearrangements.Conclusions:FISH achieved satisfactory concordance with NGS,has potential value for FGFR2 screening for targeted therapies.FGFR2 detection should be prioritized for unique clinical subgroups in ICC,which features a histological small duct subtype,early clinical stage,and reduced mucus production.

New Algorithm Rules Out Acute-on-chronic Liver Failure Development within 28 Days from Acute Decompensation of Cirrhosis

Background and Aims:Approximately 10%of patients with acute decompensated(AD)cirrhosis develop acute-on-chronic liver failure(ACLF)within 28 days.Such cases have high mortality and are difficult to predict.Therefore,we aimed to establish and validate an algorithm to identify these patients on hospitalization.Methods:Hospitalized patients with AD who developed ACLF within 28 days were considered pre-ACLF.Organ dysfunction was defined accord-ing to the chronic liver failure-sequential organ failure as-sessment(CLIF-SOFA)criteria,and proven bacterial infec-tion was taken to indicate immune system dysfunction.A retrospective multicenter cohort and prospective one were used to derive and to validate the potential algorithm,re-spectively.A miss rate of<5%was acceptable for the calcu-lating algorithm to rule out pre-ACLF.Results:In the deri-vation cohort(n=673),46 patients developed ACLF within 28 days.Serum total bilirubin,creatinine,international normalized ratio,and present proven bacterial infection at admission were associated with the development of ACLF.AD patients with≥2 organ dysfunctions had a higher risk for pre-ACLF patients[odds ratio=16.58195%confidence interval:(4.271-64.363),p<0.001].In the derivation co-hort,67.5%of patients(454/673)had≤1 organ dysfunction and two patients(0.4%)were pre-ACLF,with a miss rate of 4.3%(missed/total,2/46).In the validation cohort,65.9%of patients(914/1388)had≤1 organ dysfunction,and four(0.3%)of them were pre-ACLF,with a miss rate of 3.4%(missed/total,4/117).Conclusions:AD patients with≤1 organ dysfunction had a significantly lower risk of developing ACLF within 28 days of admission and could be safely ruled out with a pre-ACLF miss rate of<5%.

Investigation on the short-term outcome and prognostic impact of predisposition,and precipitants in inpatients with chronic liver disease from Chinese AcuTe on CHronic LIver FailurE(CATCH-LIFE)cohorts

Aim:The study aimed to investigate the short-term outcomes of hospitalized patients with chronic liver disease(CLDs)and assess the prognostic impact of predisposition and precipitants,which currently remains unclear.Methods:The study included 3970 hospitalized patients with CLDs from two prospective longitudinal multicenter studies(NCT02457637 and NCT03641872)conducted in highly endemic hepatitis B virus(HBV)areas.Competing risk analysis was used to evaluate the effect of predispositions,including the etiology and severity of CLDs and precipitants;on sequential 28,90,and 365-day liver transplantation(LT)-free mortality.Results:Among all enrolled patients,76.8%of adverse outcomes(including death and LT)within one year occurred within 90 days.Compared with alcoholic etiology,the association of HBV etiology with poorer outcomes was remarkably on the 28th day(hazard ratio[HR],1.81;95%confidence interval[CI],1.07-3.06;p=0.026);however,and dimin-ished or became insignificant at 90 days and 365 days.Cirrhosis increased the adjusted risk for 365-day(HR,1.50;CI,1.13-1.99;p=0.004)LT-free mortality when compared with noncirrhosis.In patients with cirrhosis,prior decompensation(PD)independently increased the adjusted risk of 365-day LT-free mortality by 1.25-fold(p=0.021);however,it did not increase the risk for 90-day mortality.Neither the category nor the number of precipitants influenced the adjusted risk of 28 or 90-day LT-free mortality.Conclusions:The 90-day outcome should be considered a significant endpoint for evaluating the short-term prognosis of hospitalized patients with CLD.Predisposing factors,other than etiology,mainly affected the delayed(365-day)outcome.Timely effective therapy for CLD etiology,especially antiviral treatments for HBV,and post-discharge long-term surveillance monitoring in cirrhotic patients undergoing PD are suggested to enhance disease management and reduce mortality.

Amine-functionalized SiO2 nanodot-coated layered double hydroxide nanocomposites for enhanced gene delivery

在这个工作，新奇策略被开发了准备分散得好的 amine-functionalized SiO <潜水艇class=“ a-plus-plus ”> 2 nanodot涂的分层的双氢氧化物 nanocomposite ( NH <潜水艇class=“ a-plus-plus ”> 2 -SiO经由静电的相互作用和冷凝作用的 2 @LDH)( 3-aminopropyl ) triethoxysilane ( APTES )。这个 nanocomposite 系统很好在缓冲的文化媒介和磷酸盐被驱散盐，并且展出低 cytotoxicity 和好 biocompatibility。荧光显微镜学图象和流动 cytometry 数据显示那如此的 NH < 潜水艇 class= “ a-plus-plus ” > 2 -SiO 2 @LDH nanocomposite 能高效地交付小介入的 RNA (siRNA ) 进 U2OS 房间线禁止房间增长。因此， NH < 潜水艇 class= “ a-plus-plus ” > 2 -SiO 2 @LDH nanocomposite 为有效基因交货作为 nanocarrier 有一个大潜力。

Size-dependent gene delivery of amine-modified silica nanoparticles

Silica-based nanoparticles are promising carriers for gene delivery applications. To gain insights into the effect of particle size on gene transfection efficiency, amine-modified monodisperse St6ber spheres （NH2-SS） with diameters of 125, 230, 330, 440, and 570 nm were synthesized. The in vitro transfection efficiencies of NH2-SS for delivering plasmid DNA encoding green fluorescent protein （GFP） （pcDNA3-EGFP, abbreviated as pcDNA, 6.1 kbp） were studied in HEK293T cells. NH2-SS with a diameter of 330 nm （NH2-SS330） showed the highest GFP transfection level compared to NH2-SS particles with other sizes. The transfection efficiency was found as a compromise between the binding capacity and cellular uptake performance of NH2-SS330 and pcDNA conjugates. NH2-SS330 also demonstrated the highest transfection efficiency for plasmid DNA （pDNA） with a bigger size of 8.9 kbp. To our knowledge, this study is the first to demonstrate the significance of particle size for gene transfection efficiency in silica-based gene delivery systems. Our findings are crucial to the rational design of synthetic vectors for gene therapy.