Reproducibility of the Nottingham modification of the Scarff- Bloom-Richardson histological grading system and the complementary value of Ki-67 to this system

Background The reproducibility of the Nottingham modification of the Scarff-Bloom-Richardson （NSBR） histological grading system for invasive breast cancer （IBC） adopted by the World Health Organization （WHO） has previously not been studied in Chinese hospitals. The proliferation marker, Ki-67, has been widely applied in detecting IBC. The objective of this study was to assess the reproducibility of the NSBR system among Chinese pathologists and the complementary value that Ki-67 brings to this system.Methods Four general pathologists graded 100 IBC cases independently, which had previously been graded by specialists in breast pathology. The interobserver reproducibility among four general pathologists and pairwise reproducibility between each of them and the specialists were assessed. The Ki-67 labeling index （Ki-67LI） was determined by immunohistochemistry, and its correlations with histological grade and survival were determined.Results With respect to interobserver reproducibility, NSBR grading was fairly reproducible （kappa=0.34）; as for the components of NSBR grading, agreement was best for tubule formation （kappa=0.46）, intermediate for nuclear pleomorphism （kappa=0.42）, and poorest for mitotic count （kappa=0.28）. In terms of pairwise reproducibility, agreement was fair to substantial with NSBR grading （kappa=0.30-0.69） and nuclear pleomorphism （kappa=0.28-0.69）, moderate to substantial for tubule formation （kappa=0.51-0.78）, and slight to substantial for mitotic count （kappa=0.19-0.71）.There were characteristic Ki-67LI ranges for grades 1,2 and 3 tumors. Univariate analysis showed that Ki-67 was able to divide grade 2 patients into two different prognostic subgroups. Multivariate analysis of grade 2 patients with negative lymph node demonstrated that Ki-67 was an independent prognosticator for overall survival.Conclusions The reproducibility of grading by general pathologists could be enhanced. Specialization in breast pathology is essential for accurate grading and treatment for IBC. Ki-67, with proven prognostic significance, adds complementary value to the NSBR system.

Low-depth whole genome sequencing reveals copy number variations associated with higher pathologic grading and more aggressive subtypes of lung non-mucinous adenocarcinoma

Objective:Histology grade,subtypes and TNM stage of lung adenocarcinomas are useful predictors of prognosis and survival.The aim of the study was to investigate the relationship between chromosomal instability,morphological subtypes and the grading system used in lung non-mucinous adenocarcinoma(LNMA).Methods:We developed a whole genome copy number variation(WGCNV)scoring system and applied next generation sequencing to evaluate CNVs present in 91 LNMA tumor samples.Results:Higher histological grades,aggressive subtypes and more advanced TNM staging were associated with an increased WGCNV score,particularly in CNV regions enriched for tumor suppressor genes and oncogenes.In addition,we demonstrate that 24-chromosome CNV profiling can be performed reliably from specific cell types(<100 cells)isolated by sample laser capture microdissection.Conclusions:Our findings suggest that the WGCNV scoring system we developed may have potential value as an adjunct test for predicting the prognosis of patients diagnosed with LNMA.

Gastroesophageal reflux following per-oral endoscopic myotomy:Can we improve outcomes?

This editorial is an analysis the review article by Nabi et al recently published in this journal.Achalasia Cardia is a disease whose pathophysiology is still unclear.It is known that there is inflammation of unknown aetiology leading to loss of ganglion cells in the muscularis propria.The end result is lower oesophageal sphincter spasm,loss of receptive relaxation,decreased oesophageal peristalsis,all leading on to varying degrees of dysphagia.The treatment of this condition is palliative in nature,performed by myotomy of the lower oesophagus either surgically or endoscopically.Gastroesophageal reflux disease(GERD)has been associated with the myotomy performed,particularly with the Peroral Endoscopic Myotomy(POEM)procedure.Nabi et al have provided an excellent overview of the latest developments in predicting,preventing,evaluating,and managing GERD subsequent to POEM.Based on this theme,this review article explores the concept of using histology of the oesophageal muscle layer,to grade the disease and thereby help tailoring the length/type of myotomy performed during the POEM procedure.In the future,will a histology based algorithm available preoperatively,help modify the POEM procedure,thereby decreasing the incidence of GERD associated with POEM?

RNF13： a novel RING-type ubiquitin ligase over-expressed in pancreatic cancer

Protein ubiquitination by E3 ubiquitin ligases plays an important role in cancer development. In this study, we provide experimental evidence that a RING-finger-containing protein RNF13 is an ER/Golgi membrane-associated E3 ubiquitin ligase and its RING finger domain is required for the ubiquitin ligase activity. Immunohistochemical analysis of pancreatic ductal adenocarcinoma （PDAC） and paracancerous normal tissues from 72 patients documented RNF13 over-expression in 30 tumor samples （41.7%, 30/72）, and its expression was significantly associated with histological grading （P= 0.024）. In addition, RNF13 was detected in precancerous lesions： tubular complexes in chronic pancreatitis （CP） and pancreatic intraepithelial neoplasia （PanIN） （79.3%, 23/29 and 62.8%, 22/35, respectively）. Moreover, RNF13 staining was significantly correlated with Tenascin-C expression （P = 0.004） in PDAC samples, further supporting the role of RNF13 in cancer progression. Over-expression of wild type but not RING domain-mutant RNF13 in pancreatic MiaPaca-2 cancer cells increased invasive potential and gelatinolytic activity by matrix metalloproteinase-9. Taken together, these findings reveal that RNF13 is a novel E3 ubiquitin ligase involved in pancreatic carcinogenesis; ubiqui-tin-mediated modification of proteins by RNF13 may participate in pancreatic cancer development.

Clinicopathological features and prognosis assessment of extranodal follicular dendritic cell sarcoma

AIM: To establish a model for prognosis assessment of extranodal follicular dendritic cell (FDC) sarcoma.METHODS: Nine lesions were examined by routine and molecular approaches.Clinicopathological factors from the new cases and 97 reported cases were analyzed for their prognostic values.RESULTS: The current lesions were found in f ive male and four female patients,located mainly in the head and neck area and averaging 7.2 cm in size.Six patients had recurrence or metastasis and three remained free of disease.The 106 patients (male/female ratio,1.1:1) were aged from 9 to 82 years (median,44 years).The tumor sizes ranged from 1.5 to 21 cm (mean,7.4 cm).Abdominal/pelvic region was affected most frequently (43%).Surgical resection was performed in 100 patients,followed by radiation and/or chemotherapy in 35 of them.Follow-up data were available in 91 cases,covering a period of 3-324 mo (mean,27 mo;median,19 mo).Of the informative cases,38 (42%) had recurrence or metastasis,and 12 (13%) died of the disease.These tumors were classif ied histologically into lowand high-grade lesions.A size ≥ 5 cm (P = 0.003),highgrade histology (P = 0.046) and a mitotic count ≥ 5/10 HPF (P = 0.013) were associated with tumor recurrence.The lesions were def ined as low-,intermediateand high-risk tumors,and their recurrence rates were 16%,46% and 73%,and their mortality rates 0%,4% and 45%,respectively.CONCLUSION: Extranodal FDC tumors behave like soft tissue sarcomas.Their clinical outcomes are variable and can be evaluated according to their sizes and grades.

Shear wave elastography results correlate with liver fibrosis histology and liver function reserve

AIM: To evaluate the correlation of shear wave elastography (SWE) results with liver fibrosis histology and quantitative function reserve. METHODS: Weekly subcutaneous injection of 60% carbon tetrachloride (1.5 mL/kg) was given to 12 canines for 24 wk to induce experimental liver fibrosis, with olive oil given to 2 control canines. At 24 wk, liver condition was evaluated using clinical biochemistry assays, SWE imaging, lidocaine metabolite monoethylglycine-xylidide (MEGX) test, and histologic fibrosis grading. Clinical biochemistry assays were performed at the institutional central laboratory for routine liver function evaluation. Liver stiffness was measured in triplicate from three different intercostal spaces and expressed as mean liver stiffness modulus (LSM). Plasma concentrations of lidocaine and its metabolite MEGX were determined using high-performance liquid chromatography repeated in duplicate. Liver biopsy samples were fixed in 10% formaldehyde, and liver fibrosis was graded using the modified histological activity index Knodell score (F0-F4). Correlations among histologic grading, LSM, and MEGX measures were analyzed with the Pearson linear correlation coefficient. RESULTS: At 24 wk liver fibrosis histologic grading was as follows: F0, n = 2 (control); F1, n = 0; F2, n = 3; F3, n = 7; and F4, n = 2. SWE LSM was positively correlated with histologic grading (r = 0.835, P < 0.001). Specifically, the F4 group had a significantly higher elastic modulus than the F3, F2, and F0 groups (P = 0.002, P = 0.003, and P = 0.006, respectively), and the F3 group also had a significantly higher modulus than the control F0 group (P = 0.039). LSM was negatively associated with plasma MEGX concentrations at 30 min (r = -0.642; P = 0.013) and 60 min (r = -0.651; P = 0.012), time to 1/2 of the maximum concentration (r = -0.538; P = 0.047), and the area under the curve (r = -0.636; P = 0.014). Multiple comparisons showed identical differences in these three measures: significantly lower with F4 (P = 0.037) and F3 (P = 0.032) as compared to F0 and significantly lower with F4 as compared to F2 (P = 0.032). CONCLUSION: SWE LSM shows a good correlation with histologic fibrosis grading and pharmacologic quantitative liver function reserve in experimental severe fibrosis and cirrhosis.

Development and validation of a CT-based radiomics nomogram for preoperative prediction of tumor histologic grade in gastric adenocarcinoma

Objectives:To develop and validate a radiomics nomogram for preoperative prediction of tumor histologic grade in gastric adenocarcinoma(GA).Methods:This retrospective study enrolled 592 patients with clinicopathologically confirmed GA(low-grade:n=154;high-grade:n=438)from January 2008 to March 2018 who were divided into training(n=450)and validation(n=142)sets according to the time of computed tomography(CT)examination.Radiomic features were extracted from the portal venous phase CT images.The Mann-Whitney U test and the least absolute shrinkage and selection operator(LASSO)regression model were used for feature selection,data dimension reduction and radiomics signature construction.Multivariable logistic regression analysis was applied to develop the prediction model.The radiomics signature and independent clinicopathologic risk factors were incorporated and presented as a radiomics nomogram.The performance of the nomogram was assessed with respect to its calibration and discrimination.Results:A radiomics signature containing 12 selected features was significantly associated with the histologic grade of GA(P<0.001 for both training and validation sets).A nomogram including the radiomics signature and tumor location as predictors was developed.The model showed both good calibration and good discrimination,in which C-index in the training set,0.752[95%confidence interval(95%CI):0.701-0.803];C-index in the validation set,0.793(95%CI:0.711-0.874).Conclusions:This study developed a radiomics nomogram that incorporates tumor location and radiomics signatures,which can be useful in facilitating preoperative individualized prediction of histologic grade of GA.

Serum neopterin levels in children with hepatitis-B-related chronic liver disease and its relationship to disease severity

AIM:To evaluate serum neopterin levels and their correlations with liver function tests and histological grade in children with hepatitis-B-related chronic liver disease. METHODS:The study population comprised 48 patients with chronic active hepatitis B,32 patients with hepatitis-B-related active liver cirrhosis and 40 normal controls. Serum neopterin was measured using an enzyme-linked immunosorbent assay. RESULTS:The mean ± SD serum neopterin levels were 14.2 ± 5.6 nmol/L in patients with chronic hepatitis,20.3 ± 7.9 nmol/L in patients with liver cirrhosis and 5.2 ± 1.4 nmol/L in control group. Serum neopterin levels were signif icantly higher in patients with chronic hepatitis (P = 0.005) and cirrhosis patients (P = 0.008),than in control subjects. Cirrhotic patients had signif icantly higher serum neopterin levels than patients with chronic hepatitis (P = 0.004). There was a positive correlation between serum neopterin levels and alanine aminotransferase levels in patients with chronic hepatitis (r = 0.41,P = 0.004) and cirrhotic patients (r = 0.39,P = 0.005). Positive correlations were detected between serum neopterin levels and inflammatory score in patients with chronic hepatitis (r = 0.51,P = 0.003) and cirrhotic patients (r = 0.49,P = 0.001). CONCLUSION:Our results suggest that serum neopterin levels can be considered as a marker of inflammatory activity and severity of disease in children with hepatitis-B-related chronic liver disease.

Low-Grade and High-Grade Invasive Ductal Carcinomas of the Breast Follow Divergent routes of Progression

Low-grade invasive ductal carcinoma is almost diploid, and has frequent losses of chromosome 16q, which is shared by other precancerous lesions of the mammary gland such as flat epithelial atypia （FEA）, atypical ductal hyperplasia （ADH）, and lownuclear grade ductal carcinoma in situ （DCIS）. The genetic alterations accumulate in a stepwise fashion as the precancerous lesions progress to invasve ductal carcinoma. This supports the linear progression model of breast cancer from FEA, through ADH, to low- nuclear grade DCIS as non-obligate early events in low-grade IDC evolution. In contrast, high-grade carcinoma tends to aneuploidy with complex genetic alterations--most importantly, frequent gains at chromosome 16q. Frequent losses at chromosome 16q in low-grade IDC and gains in the same arm of the same chromosome in high-grade IDC imply that these lesions are two end outcomes of different disease processes and that they do not lie in the same continuum of a process. Therefore, low-grade and high-grade IDC are two distinct diseases with a divergent route of progression.

INVESTIGATION OF RELATIONSHIPS BETWEEN KI－67SCORE，DNA INDEX，AND HISTOLOGIC GRADEIN SOFT TISSUE SARCOMAS

Proliferative activity of soft tissue sarcomas(STS)in 31 cases was estimated by histologic grading, mitotic count, DNA analysis by flow cytometry, and immunohistochemical procedures with monoclonal antibody Ki-67.Aneuploid was found in 12 of 16 cases(75.0%) with Grade 3, and in 4 of 15 cases(26.7%) with Grade 1, 2(P=0.0121).Tumors with more than 100 Ki-67 positive tumor cells per 10 high power fields(HPF) had a higher rate of aneuploid(81.3%) than those with less than 100 Ki-67 per 10 HPF (26.7%)(P=0.0038). There were significant correlations between Grade and DI (r=0.4901, P=0.0051), Grade and Ki-67(r=0.4636, P=0.0086), Ki-67 and DI(r=0-6368, P=0.0001). The results indicate that DI and reactivity of tumor cells to Ki-67 may reflect Proliferative activity and be helpful for clinicians to judge the biological behaviour of tumors more accurately and objectively. Supplementary to the grading of STS, DI and Ki-67 score could be useful as Prognostic parameters for clinical investigation of multimodality therapy for individual Patients.

HISTOLOGIC GRADE, AND CONTENT AND IMAGE ANALYSIS OF SOFT TISSUE TUMORS

Flow cytometry and image analysis technique were used to quantltate the nuclei of various soft tissue tumors. A single representing section from soft tissue sarcoma was used for histologic grading. Histologlc and cytometric comparative analyses showed that all 21 benign tumors were diploid. Among 62 cases of soft tissue sarcoma, 45(73%) were aneuploid. There was a significant difference in the nuclear area between benign and malignant tumors (P<0. 01), dlploid and aneuploid tumors (P<0. 05). The two new techniques are valuable In cellular quantitative measurement for soft tissue tumors.

Value of perfusion parameters and histogram analysis of triphasic computed tomography in pre-operative prediction of histological grade of hepatocellular carcinoma

Background:Pre-operative non-invasive histological evaluation of hepatocellular carcinoma(HCC)remains a challenge.Tumor perfusion is significantly associated with the development and aggressiveness of HCC.The purpose of the study was to evaluate the clinical value of quantitative liver perfusion parameters and corresponding histogram parameters derived from traditional triphasic enhanced computed tomography(CT)scans in predicting histological grade of HCC.Methods:Totally,52 patients with HCC were enrolled in this retrospective study and underwent triple-phase enhanced CT imaging.The blood perfusion parameters were derived from triple-phase CT scans.The relationship of liver perfusion parameters and corresponding histogram parameters with the histological grade of HCC was analyzed.Receiver operating characteristic(ROC)curve analysis was used to determine the optimal ability of the parameters to predict the tumor histological grade.Results:The variance of arterial enhancement fraction(AEF)was significantly higher in HCCs without poorly differentiated components(NP-HCCs)than in HCCs with poorly differentiated components(P-HCCs).The difference in hepatic blood flow(HF)between total tumor and total liver flow(ΔHF=HFtumor-HFliver)and relative flow(rHF=ΔHF/HF_(liver))were significantly higher in NP-HCCs than in P-HCCs.The difference in portal vein blood supply perfusion(PVP)between tumor and liver tissue(ΔPVP)and theΔPVP/liver PVP ratio(rPVP)were significantly higher in patients with NP-HCCs than in patients with P-HCCs.The area under ROC(AUC)ofΔPVP and rPVP were both 0.697 with a high sensitivity of 84.2%and specificity of only 56.2%.TheΔHF and rHF had a higher specificity of 87.5%with an AUC of 0.681 and 0.673,respectively.The combination of rHF and rPVP showed the highest AUC of 0.732 with a sensitivity of 57.9%and specificity of 93.8%.The combined parameter ofΔHF and rPVP,rHF and rPVP had the highest positive predictive value of 0.903,and that of rPVP andΔPVP had the highest negative predictive value of 0.781.Conclusion:Liver perfusion parameters and corresponding histogram parameters(includingΔHF,rHF,ΔPVP,rPVP,and AEFvariance)in patients with HCC derived from traditional triphasic CT scans may be helpful to non-invasively and pre-operatively predict the degree of the differentiation of HCC.

Gd-EOB-DTPA-enhanced MRI radiomic features for predicting histological grade of hepatocellular carcinoma

Background:Prediction models for the histological grade of hepatocellular carcinoma(HCC)remain unsatisfactory.The purpose of this study is to develop preoperative models to predict histological grade of HCC based on gadolinium-ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)-enhanced magnetic resonance imaging(MRI)radiomics.And to compare the performance between artificial neural network(ANN)and logistic regression model.Methods:A total of 122 HCCs were randomly assigned to the training set(n=85)and the test set(n=37).There were 242 radiomic features extracted from volumetric of interest(VOI)of arterial and hepatobiliary phases images.The radiomic features and clinical parameters[gender,age,alpha-fetoprotein(AFP),carcinoembryonic antigen(CEA),carbohydrate antigen 19-9(CA19-9),alanine aminotransferase(ALT),aspartate transaminase(AST)]were selected by permutation test and decision tree.ANN of arterial phase(ANN-AP),logistic regression model of arterial phase(LR-AP),ANN of hepatobiliary phase(ANN-HBP),logistic regression mode of hepatobiliary phase(LR-HBP),ANN of combined arterial and hepatobiliary phases(ANN-AP+HBP),and logistic regression model of combined arterial and hepatobiliary phases(LR-AP+HBP)were built to predict HCC histological grade.Those prediction models were assessed and compared.Results:ANN-AP and LR-AP were composed by AST and radiomic features based on arterial phase.ANN-HBP and LR-HBP were composed by AFP and radiomic features based on hepatobiliary phase.ANN-AP+HBP and LR-AP+HBP were composed by AST and radiomic features based on arterial and hepatobiliary phases.The prediction models could distinguish between high-grade tumors[Edmondson-Steiner(E-S)grade III and IV]and low-grade tumors(E-S grade I and II)in both training set and test set.In the test set,the AUCs of ANN-AP,LR-AP,ANN-HBP,LR-HBP,ANN-AP+HBP and LR-AP+HBP were 0.889,0.777,0.941,0.819,0.944 and 0.792 respectively.The ANN-HBP was significantly superior to LR-HBP(P=0.001).And the ANN-AP+HBP was significantly superior to LR-AP+HBP(P=0.007).Conclusions:Prediction models consisting of clinical parameters and Gd-EOB-DTPA-enhanced MRI radiomic features(based on arterial phase,hepatobiliary phase,and combined arterial and hepatobiliary phases)could distinguish between high-grade HCCs and low-grade HCCs.And the ANN was superior to logistic regression model in predicting histological grade of HCC.

Ki-67 index,progesterone receptor expression,histologic grade and tumor size in predicting breast cancer recurrence risk:A consecutive cohort study

Background:The 21-gene recurrence score(RS)assay has been recommended by major guidelines for treatment decision in hormone receptor(HR)-positive early breast cancer(EBC).However,the genomic assay is not accessible and affordable worldwide.Alternatively,an increasing number of studies have shown that traditional immunohistochemistry(IHC)can partially or even completely replace the role of the 21-gene genomic assay.Here,we developed and validated a predictive model(IHC3 model)combining the Ki-67 index,progesterone receptor(PR)expression,histologic grade,and tumor size to predict the recurrence risk of HR-positive EBC.Methods:The data from 389 patients(development set)with HR-positive,human epidermal growth factor receptor 2-negative,lymph node non-metastasized invasive breast cancer were used to construct the IHC3 model based on the Surexam®21-gene RS and the TAILORx clinical trial criteria.An additional 146 patients with the same characteristics constituted the validation set.The predictive accuracy of the IHC3 model was compared with that of Orucevic et al.’s nomogram.Invasive diseasefree survival(IDFS)was analyzed in the IHC3 predictive low-recurrence risk(pLR)group and the predictive high-recurrence risk(pHR)group.The Pearson chi-square test,Fisher exact test,and log-rank test were used for analysis.Results:The pLR and pHR group could be easily stratified using the decision tree model without network dependence.The accuracies of the IHC3 model were 86.1%in the development set and 87.7%in the validation set.The predictive accuracy of the IHC3 model and Orucevic et al.’s nomogram for the whole cohort was 86.5%and 86.9%,respectively.After a 52-month of median follow-up,a significant difference was found in IDFS between of the IHC3 pLR and the pHR groups(P=0.001)but not in the IDFS between the low-and high-recurrence risk groups according to the Surexam®21-gene RS and the TAILORx clinical trial criteria(P=0.556)or 21-gene binary RS group(P=0.511).Conclusions:The proposed IHC3 model could reliably predict low and high recurrence risks in most HR-positive EBC patients.This easy-to-use predictive model may be a reliable replacement for the 21-gene genomic assay in patients with EBC who have no access to or cannot afford the 21-gene genomic assay.