Radiomics signature:A potential biomarker forβ-arrestin1 phosphorylation prediction in hepatocellular carcinoma

BACKGROUND The phosphorylation status ofβ-arrestin1 influences its function as a signal strongly related to sorafenib resistance.This retrospective study aimed to develop and validate radiomics-based models for predictingβ-arrestin1 phosphorylation in hepatocellular carcinoma(HCC)using whole-lesion radiomics and visual imaging features on preoperative contrast-enhanced computed tomography(CT)images.AIM To develop and validate radiomics-based models for predictingβ-arrestin1 phosphorylation in HCC using radiomics with contrast-enhanced CT.METHODS Ninety-nine HCC patients(training cohort:n=69;validation cohort:n=30)receiving systemic sorafenib treatment after surgery were enrolled in this retrospective study.Three-dimensional whole-lesion regions of interest were manually delineated along the tumor margins on portal venous CT images.Radiomics features were generated and selected to build a radiomics score using logistic regression analysis.Imaging features were evaluated by two radiologists independently.All these features were combined to establish clinico-radiological(CR)and clinico-radiological-radiomics(CRR)models by using multivariable logistic regression analysis.The diagnostic performance and clinical usefulness of the models were measured by receiver operating characteristic and decision curves,and the area under the curve(AUC)was determined.Their association with prognosis was evaluated using the Kaplan-Meier method.RESULTS Four radiomics features were selected to construct the radiomics score.In the multivariate analysis,alanine aminotransferase level,tumor size and tumor margin on portal venous phase images were found to be significant independent factors for predictingβ-arrestin1 phosphorylation-positive HCC and were included in the CR model.The CRR model integrating the radiomics score with clinico-radiological risk factors showed better discriminative performance(AUC=0.898,95%CI,0.820 to 0.977)than the CR model(AUC=0.794,95%CI,0.686 to 0.901;P=0.011),with increased clinical usefulness confirmed in both the training and validation cohorts using decision curve analysis.The risk ofβ-arrestin1 phosphorylation predicted by the CRR model was significantly associated with overall survival in the training and validation cohorts(log-rank test,P<0.05).CONCLUSION The radiomics signature is a reliable tool for evaluatingβ-arrestin1 phosphorylation which has prognostic significance for HCC patients,providing the potential to better identify patients who would benefit from sorafenib treatment.

A Macaca mulatta model of fulminant hepatic failure

AIM:To establish an appropriate primate model of fulminant hepatic failure (FHF).METHODS:We have,for the first time,established a large animal model of FHF in Macaca mulatta by intraperitoneal infusion of amatoxin and endotoxin.Clinical features,biochemical indexes,histopathology and iconography were examined to dynamically investigate the progress and outcome of the animal model.RESULTS:Our results showed that the enzymes and serum bilirubin were markedly increased and the enzyme-bilirubin segregation emerged 36 h after toxin administration.Coagulation activity was significantly decreased.Gradually deteriorated parenchymal abnormality was detected by magnetic resonance imaging (MRI) and ultrasonography at 48 h.The liver biopsy showed marked hepatocyte steatosis and massive parenchymal necrosis at 36 h and 49 h,respectively.The autopsy showed typical yellow atrophy of the liver.Hepatic encephalopathy of the models was also confirmed by hepatic coma,MRI and pathological changes of cerebral edema.The lethal effects of the extrahepatic organ dysfunction were ruled out by their biochemical indices,imaging and histopathology.CONCLUSION:We have established an appropriate large primate model of FHF,which is closely similar to clinic cases,and can be used for investigation of the mechanism of FHF and for evaluation of potential medical therapies.

Proportions of acetyl-histone-positive hepatocytes indicate the functional status and prognosis of cirrhotic patients

AIM:To investigate whether the proportions of acetylhistone-positive hepatocytes could be used as markers of deteriorating liver function.METHODS:In total,611 cirrhotic cases from 3701 patients who were diagnosed during the past 15 years were screened,and 152 follow-up cases were selected.Paraffin tissue microarray was prepared for immunohistochemistry to examine acetyl-histone expression.The proportions of positive hepatocytes were recorded,and their correlations to clinical and laboratory indicators were analyzed statistically.RESULTS:The proportions of H2AK5ac+,H3K9/K14ac+ and H3K27ac+ hepatocytes gradually increased with deteriorating liver function and with increasing levels of serum markers of liver injury.In the follow-up cases,patients with > 70% H2AK5ac+,H3K9/K14ac+ or H3K27ac+ hepatocytes had statistically lower survival rates(P < 0.05).Furthermore,> 70% H2AK5ac+ or H3K27ac+ hepatocytes were strong independent predictors of overall survival(P < 0.05).CONCLUSION:The proportions of acetyl-histonepositive hepatocytes are closely associated with the liver function and prognosis of cirrhotic patients.

Mononuclear macrophages in pathogenesis of acute lung injury during acute obstructive cholangitis

Objective: To determine the role of mononuclear macrophages in the pathogenesis of acute lung injury during acute obstructive cholangitis. Methods: Sixty Wistar rats were used to study the correlation between the behavior of mononuclear macrophages and acute pulmonary injury during a- cute obstructive cholangitis (AOC). Animal model of AOC was made according to the method that the common bile duct was injected with Escherichia coli and ligated. The rats were killed at 6 h, 12 h, 24 h and 48 h after operation. The phagocytic function of Kupffer cells (KCs), the number of alveolar macro- phages (AMs) in bronchoalveolar lavage liquid, and the extravascular water content of lung tissue were measured. The levels of lipid peroxide (LPO) and supperoxide dismutase (SOD) were determined too. Pathological alterations of liver and lung tissue were observed under light and electron microscopes. Results: KCs phagocytic function was significantly el- evated at the 6th hour but markedly decreased from the 24th hour to the 48th hour in the AOC group as compared with the control (P<0. 05). From the 12th to the 48th hour, the number of AMs, the ex- travascular water content of lung tissue, and the con- tent of LPO significantly increased, but the SOD lev- el of lung tissue decreased greatly (P<0. 05). Mor- phologically, KCs proliferated diffusely in the early period in livers of the AOC group, but decreased markedly in the late period. Mitochondria of KCs were swollen or even vacuolated; focal cytoplasmic degeneration and many myeli like figures could be seen in the cytoplasm. The changes of injury such as disturbance of pulmonary capillary blood circula- tion, degeneration and/or necrosis of the lung tissue and endothelium, and inflammatory reactions could be observed. In other two groups, no evident mor- phological changes were observed. Conclusions: KCs phagocytic function is decreased, whereas AM is activated by the invading bacteria to release such inflammatory mediators as free radicals, resulting in acute pulmonary injury. It seems that there is a close relationship between the functional status of mononuclear macrophages and the develop- ment of acute lung injury. The dysfunction of mono- nuclear macrophages may play an important role in the pathogenesis of multiple organ damage, especial- ly acute pulmonary injury.

Standard liver weight model in adult deceased donors with fatty liver: A prospective cohort study

BACKGROUND Standard liver weight(SLW)is frequently used in deceased donor liver transplantation to avoid size mismatches with the recipient.However,some deceased donors(DDs)have fatty liver(FL).A few studies have reported that FL could impact liver size.To the best of our knowledge,there are no relevant SLW models for predicting liver size.AIM To demonstrate the relationship between FL and total liver weight(TLW)in detail and present a related SLW formula.METHODS We prospectively enrolled 212 adult DDs from West China Hospital of Sichuan University from June 2019 to February 2021,recorded their basic information,such as sex,age,body height(BH)and body weight(BW),and performed abdominal ultrasound(US)and pathological biopsy(PB).The chi-square test and kappa consistency score were used to assess the consistency in terms of FL diagnosed by US relative to PB.Simple linear regression analysis was used to explore the variables related to TLW.Multiple linear regression analysis was used to formulate SLW models,and the root mean standard error and interclass correlation coefficient were used to test the fitting efficiency and accuracy of the model,respectively.Furthermore,the optimal formula was compared with previous formulas.RESULTS Approximately 28.8%of DDs had FL.US had a high diagnostic ability(sensitivity and specificity were 86.2%and 92.9%,respectively;kappa value was 0.70,P<0.001)for livers with more than a 5%fatty change.Simple linear regression analysis showed that sex(R2,0.226;P<0.001),BH(R2,0.241;P<0.001),BW(R2,0.441;P<0.001),BMI(R2,0.224;P<0.001),BSA(R2,0.454;P<0.001)and FL(R2,0.130;P<0.001)significantly impacted TLW.In addition,multiple linear regression analysis showed that there was no significant difference in liver weight between the DDs with no steatosis and those with steatosis within 5%.Furthermore,in the context of hepatic steatosis,TLW increased positively(nonlinear);compared with the TLW of the non-FL group,the TLW of the groups with hepatic steatosis within 5%,between 5%and 20%and more than 20%increased by 0 g,90 g,and 340 g,respectively.A novel formula,namely,-348.6+(110.7 x Sex[0=Female,1=Male])+958.0 x BSA+(179.8 x FLUS[0=No,1=Yes]),where FL was diagnosed by US,was more convenient and accurate than any other formula for predicting SLW.CONCLUSION FL is positively correlated with TLW.The novel formula deduced using sex,BSA and FLUS is the optimal formula for predicting SLW in adult DDs.

Decreased expression of HDAC8 indicates poor prognosis in patients with intrahepatic cholangiocarcinoma

Background: Intrahepatic cholangiocarcinoma (ICC) is a highly malignant primary tumor in the liver, and the rates of incidence and mortality are rapidly increasing globally. Histone deacetylase 8 (HDAC8) is a transcriptional regulator and is associated with tumorigenesis of several tumor types. This study aimed to evaluate the correlation between HDAC8 expression and clinicopathological parameters in ICC patients. Methods: ICC tissues and corresponding nonmalignant bile duct tissues were obtained from 60 patients. HDAC8 and Ki-67 expression were evaluated by immunohistochemistry staining. HDAC8 expression and the clinicopathological features and prognosis of the patients were analyzed. The mRNA level of HDAC8 in ICC was further analyzed using data from The Cancer Genome Atlas (TCGA). Results: The expression of HDAC8 were lower in ICC tissues (39/60, 65%) than in the corresponding nonmalignant bile duct tissues (54/60, 90%)( P = 0.001). Low HDAC8 expression in ICC was significantly associated with lymph node metastases (47.6% vs. 17.9%, P = 0.015). In addition, the positive cells rate of HDAC8 was statistically and negatively correlated with the Ki-67 index in ICC lesions ( r =?0.7660, P < 0.001). Importantly, the overall survival rate and recurrence-free survival rate in ICC patients with low HDAC8 expression were lower than those with high HDAC8 expression ( P = 0.008 and P = 0.011, respectively). Conclusions: Decreased HDAC8 expression in ICC is related to poor prognosis, and HDAC8 may be an independent prognostic indicator of ICC patients after curative resection.

Elevated expression of Gsα in intrahepatic cholangiocarcinoma associates with poor prognosis

BACKGROUND:The stimulatory G protein α subunit(Gsα)plays important roles in diverse cell processes including tu morigenesis. Activating mutations in Gsα gene(GNAS) have been reported to be associated with poor prognosis in various human carcinomas. Furthermore, Gsα signaling is crucial in promoting liver regeneration by interacting with growth factor signaling, indicating that Gsα might play a promoting role in cancer development. However, little is known about the correlation between Gsα levels and clinicopathological pa rameters in intrahepatic cholangiocarcinoma(ICC). METHODS:We performed immunoblotting to examine the expression levels of Gsα and Ki67 proteins in tumor tissues and the corresponding adjacent tissues. A total of 74 pair of specimens resected from 74 ICC patients were examined. The association between Gsα levels and clinicopathological find ings and prognosis of the patients was evaluated.RESULTS:Western blotting demonstrated that the expression of Gsα was significantly higher in ICC tissues compared with that in their corresponding adjacent tissues. Gsα protein was highly expressed in about half of ICC tissues(48.6%, 36/74)while only 28.4%(21/74) of tumor adjacent tissues showed Gsα high expression(P=0.011). High Gsα expression in ICC was significantly associated with the numbers of tumor nodules(P=0.037) and lymph node metastases(P=0.010)Moreover, the level of Gsα was significantly and positivelycorrelated with Ki67 expression(P<0.001). In addition, the recurrence-free survival rate and overall survival rate in the Gsα high group were significantly lower than those in the Gsα low group(P=0.004 and P=0.005, respectively).CONCLUSIONS:High Gsα expression is correlated with poor prognosis in ICC patients. Gsα might serve as a potential prognostic indicator of ICC.

Loss of Dicer1 impairs hepatocyte survival and leads to chronic inflammation and progenitor cell activation

AIM:To investigate the continuous hepatic histopathological processes which occur in response to the loss of Dicer1.METHODS:We generated a hepatocyte-selective Dicer1 knockout mouse and observed the gradual hepatic histopathological changes in the mutant liver.Immunohistochemistry and Western blotting were performed to detect Dicer1 expression.We performed hematoxylin and eosin staining,Periodic acid-Schiff staining,Oil Red O staining,and Masson's trichrome staining to detect histological changes in Dicer1-deficient livers.Ki67 immunohistochemistry,terminal deoxynucleotidyl transferase-mediated d UTP nickend labeling assay,and Western blotting were used to determine hepatocyte proliferation and apoptosis.Serum biochemistry,cytokine assays,and flow cytometric analysis were performed to quantity liver necrosis and inflammation.Fibrogenic markers were determined by Western blotting and q PCR.CK19,CD133,and OV6 immunofluorescence were used to observe liver progenitor cells.Immunofluorescence and q PCR were performed to reveal embryonic gene expression.We also performed histological staining and Western blotting to analyze hepatocellular carcinoma(HCC) development.RESULTS:Dicer1 inactivation resulted in significant architecture disorganization and metabolism disruptionin the liver.Dicer1 disruption impaired hepatocyte survival and resulted in profound cell apoptosis and continuous necrosis.In contrast to previous reports,the mutant liver exhibited chronic inflammation and progressive fibrosis,and could not be repopulated by Dicer1-positive cells.In addition,extensive activation of hepatic progenitor cells was observed.Primary HCC was observed as early as 4 mo after birth.CONCLUSION:Hepatic loss of Dicer1 results in complex chronic pathological processes,including hepatocyte death,inflammatory infiltration,chronic fibrosis,compensatory proliferation,progenitor activation,and spontaneous hepatocarcinogenesis.

Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma(2022 version)

Intrahepatic cholangiocarcinoma(iCCA)can originate from the large bile duct group(segment bile ducts and area bile ducts),small bile duct group(septal bile ducts and interlobular bile ducts),and terminal bile duct group(bile ductules and canals of Hering)of the intrahepatic biliary tree,which can be histopathological corresponding to large duct type iCCA,small duct type iCCA and iCCA with ductal plate malformation pattern,and cholangiolocarcinoma,respectively.The challenge in pathological diagnosis of above subtypes of iCCA falls in the distinction of cellular morphologies,tissue structures,growth patterns,invasive behaviors,immunophenotypes,molecular mutations,and surgical prognoses.For these reasons,this expert consensus provides nine recommendations as a reference for standardizing and refining the diagnosis of pathological subtypes of iCCA,mainly based on the 5th edition of the World Health Organization Classification of Tumours of the Digestive System.

Osteogenic differentiation of mesenchymal stem cells promoted by overexpression of connective tissue growth factor

Objective:Large segmental bone defect repair remains a clinical and scientific challenge with increasing interest focusing on combining gene transfection with tissue engineering techniques.The aim of this study is to investigate the effect of connective tissue growth factor(CTGF) on the proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells(MSCs).Methods:A CTGF-expressing plasmid(pCTGF) was constructed and transfected into MSCs.Then expressions of bone morphogenesis-related genes,proliferation rate,alkaline phosphatase activity,and mineralization were examined to evaluate the osteogenic potential of the CTGF gene-modified MSCs.Results:Overexpression of CTGF was confirmed in pCTGF-MSCs.pCTGF transfection significantly enhanced the proliferation rates of pCTGF-MSCs(P<0.05).CTGF induced a 7.5-fold increase in cell migration over control(P<0.05).pCTGF transfection enhanced the expression of bone matrix proteins,such as bone sialo-protein,osteocalcin,and collagen type I in MSCs.The levels of alkaline phosphatase(ALP) activities of pCTGF-MSCs at the 1st and 2nd weeks were 4.0-and 3.0-fold higher than those of MSCs cultured in OS-medium,significantly higher than those of mock-MSCs and normal control MSCs(P<0.05).Overexpression of CTGF in MSCs enhanced the capability to form mineralized nodules.Conclusion:Overexpression of CTGF could improve the osteogenic differentiation ability of MSCs,and the CTGF gene-modified MSCs are potential as novel cell resources of bone tissue engineering.

Osteoinduction by Ca-P biomaterials implanted into the muscles of mice

The osteoinduction of porous biphasic calcium phosphate ceramics (BCP) has been widely reported and documented,but little research has been performed on rodent animals,e.g.,mice.In this study,we report osteoinduction in a mouse model.Thirty mice were divided into two groups.BCP materials (Sample A) and control ceramics (Sample B) were implanted into the leg muscle,respectively.Five mice in each group were killed at 15,30,and 45 d after surgery.Sample A and Sample B were harvested and used for hematoxylin and eosin (HE) staining,immunohistochemistry (IHC) staining,and Alizarin Red S staining to check bone formation in the biomaterials.Histological analysis showed that no bone tissue was formed 15 d after implantation (0/5) in either of the two groups.Newly-formed bone tissues were observed in Sample A at 30 d (5/5) and 45 d (5/5) after implantation;the average amounts of newly-formed bone tissues were approximately 5.2% and 8.6%,respectively.However,we did not see any bone tissue in Sample B until 45 d after implantation.Bone-related molecular makers such as bone morphogenesis protein-2 (BMP-2),collagen type I,and osteopontin were detected by IHC staining in Sample A 30 d after implantation.In addition,the newly-formed bone was also confirmed by Alizarin Red S staining.Because this is the report of osteoinduction in the rodent animal on which all the biotechnologies were available,our results may contribute to further mechanism research.

Synthesis and bioactivity of novel pyrazole oxime derivatives containing oxazole ring

A series of novel pyrazole oxime derivatives containing oxazole ring were designed and synthesized. The title compounds were structurally confirmed by 1H NMR, 13C NMR spectra and elemental analyses. Preliminary bioassay results showed that some of the title compounds displayed promising fungicidal activity besides insecticidal and acaricidal activity. Particularly, compound 8c exhibited potent fungicidal activity against cucumber Pseudoperonospora cubensis beyond good insecticidal activity against Aphis craccivora and Nilaparvata lugens.

Imaging biomarkers for predicting poor prognosis of hepatocellular carcinoma: a review

Hepatocellular carcinoma(HCC)is a primary malignancy of the liver with a high mortality rate.Heterogeneity is the main biological characteristic of HCC,which manifests through the different biological behaviors of each phenotype and ultimately,affects patient prognosis and treatment efficacy.Various aggressive biological behaviors are considered to be associated with the poor prognosis of HCC patients including poor differentiation,microvascular invasion,intracellular fat accumulation,invasive growth,bile duct invasion or tumor thrombosis,and tumor spread and metastasis,and have been reported as prognostic biomarkers.In addition,HCC results from multifactor synergistic damage,and various factors related to genetics,molecular pathology and immunohistochemistry such asβ-catenin,Ki67,cytokeratin-19,and epithelial cell adhesion molecule have an impact on HCC differentiation and prognosis.This article is an overview of the biological behaviors that lead to poor prognosis of HCC,and the roles of morphological and quantitative noninvasive imaging biomarkers in the evaluation and prediction of these behaviors.Some common biomarkers related to genetics,molecular pathology and immunohistochemistry are also briefly summarized.It is hoped that this review will provide clinicians and radiologists with an update on the development of liver imaging,and provide directions for the combination of radiology,genetics,molecular pathology and histopathology to better predict the prognosis of HCC patients.