Tetraspanins predict the prognosis and characterize the tumor immune microenvironment of glioblastoma.

Glioblastoma (GBM) is the most aggressive and lethal primary brain tumor. Conventional treatments have not achieved breakthroughs in improving survival. Therefore, novel molecular targets and biomarkers need to be identified. As signal transduction docks on the cell membrane, tetraspanins (TSPANs) are associated with various tumors; however, research on their role in GBM remains extremely scarce. Gene expression and clinicopathological characteristic data were obtained from GEPIA, CGGA, HPA, cBioPortal, and GSCA databases to analyze the mRNA and protein expression levels, prognostic value, clinical relevance, mutation status, and targeted drug sensitivity of TSPANs in GBM. Gene set enrichment analysis (GSEA), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used for biological process enrichment. Data from TCGA and TCIA were used to construct the tumor immune microenvironment landscape of TSPANs. Different R software algorithms were used to analyze the immune score, immune cell infiltration, and immune checkpoint correlation. Univariate and multivariate analyses were performed for TSPAN4, which had the most significant predictive prognostic value, and a nomogram model was constructed to predict individual outcomes. The expression and function of TSPAN4 were verified in vitro. TSPAN3/4/6/11/12/18/23/24/25/26/27/28/29/30/31expressions were significantly upregulated in GBM, and TSPAN3/4/6/11/18/24/25/26/29/30 were strongly correlated with prognosis. The expression of multiple TSPANs significantly correlated with 1p/19q co-deletion status, IDH mutation status, recurrence, age, and tumor grade. GSEA and GO analyses revealed the potential contribution of TSPANs in cell adhesion and migration. Immune correlation analysis revealed that TSPANs are related to the formation of the GBM tumor microenvironment (TME) and may influence immunotherapy outcomes. TSPAN4 is an independent prognostic factor and TSPAN4 knockdown has been demonstrated to strongly inhibit glioma cell proliferation, invasion, and migration in vitro. We comprehensively elaborated the prognostic value and potential role of differentially expressed TSPANs in GBM, including molecules that scientists have previously overlooked. This study provides a novel and comprehensive perspective on the pathological mechanisms of GBM and the future direction of individualized tumor immunotherapy, which may be a critical link between GBM malignant progression and TME remodeling.

Retraction: ASIC1a involves acidic microenvironment-induced activation and autophagy of pancreatic stellate cells.

[This retracts the article DOI: 10.1039/C8RA03679A.].

Retracted Article: ASIC1a involves acidic microenvironment-induced activation and autophagy of pancreatic stellate cells.

Acid-sensing ion channel 1a (ASIC1a), as a member of the proton-gated cation channel family, can be activated by low extracellular pH, and takes part in many acidity-associated physiopathological processes. However, whether ASIC1a is expressed in human pancreatic stellate cells (PSCs) and involved in acid-induced physiopathological events has not been reported yet. In this study, we investigated the expression of ASIC1a in PSCs and its possible role in the activation and autophagy of PSCs evoked by extracellular acid. Our results show that ASIC1a is present in PSCs, and an enhanced expression of ASIC1a occurs under acid stimuli. More importantly, the activation and autophagy of PSCs can be induced in acidic medium, and inhibition of ASIC1a by ASIC1a-specific blocker psalmotoxin-1 (PcTx1) or siRNA knockdown could suppress these two acid-associated processes. Collectively, our present study reports for the first time that ASIC1a is expressed in PSCs, and provides evidence for the involvement of ASIC1a in the acidic microenvironment-induced activation and autophagy of PSCs.

Cross-modality PET/CT and contrast-enhanced CT imaging for pancreatic cancer.

AIM: To explore the diagnostic value of the cross-modality fusion images provided by positron emission tomography/computed tomography (PET/CT) and contrast-enhanced CT (CECT) for pancreatic cancer (PC). METHODS: Data from 70 patients with pancreatic lesions who underwent CECT and PET/CT examinations at our hospital from August 2010 to October 2012 were analyzed. PET/CECT for the cross-modality image fusion was obtained using TureD software. The diagnostic efficiencies of PET/CT, CECT and PET/CECT were calculated and compared with each other using a χ(2) test. P < 0.05 was considered to indicate statistical significance. RESULTS: Of the total 70 patients, 50 had PC and 20 had benign lesions. The differences in the sensitivity, negative predictive value (NPV), and accuracy between CECT and PET/CECT in detecting PC were statistically significant (P < 0.05 for each). In 15 of the 31 patients with PC who underwent a surgical operation, peripancreatic vessel invasion was verified. The differences in the sensitivity, positive predictive value, NPV, and accuracy of CECT vs PET/CT and PET/CECT vs PET/CT in diagnosing peripancreatic vessel invasion were statistically significant (P < 0.05 for each). In 19 of the 31 patients with PC who underwent a surgical operation, regional lymph node metastasis was verified by postsurgical histology. There was no statistically significant difference among the three methods in detecting regional lymph node metastasis (P > 0.05 for each). In 17 of the 50 patients with PC confirmed by histology or clinical follow-up, distant metastasis was confirmed. The differences in the sensitivity and NPV between CECT and PET/CECT in detecting distant metastasis were statistically significant (P < 0.05 for each). CONCLUSION: Cross-modality image fusion of PET/CT and CECT is a convenient and effective method that can be used to diagnose and stage PC, compensating for the defects of PET/CT and CECT when they are conducted individually.

Hemodynamic alterations in unilateral chronic middle cerebral artery stenosis patients and the effect of percutaneous transluminal angioplasty and stenting: a perfusion-computed tomography study.

BACKGROUND: The Stenting and Aggressive Medical Management for Preventing Recurrent Stroke in Intracranial Stenosis (SAMMPRIS) trial indicated that most patients with symptomatic intracranial atherosclerosis are not good candidates for percutaneous transluminal angioplasty and stenting (PTAS) because of a higher complication risk than with conservative treatment. However, enrollment of SAMMPRIS patients was based on lesion severity only, without functional imaging. PURPOSE: To determine whether perfusion computed tomography (PCT) can effectively evaluate hemodynamic compromise in unilateral chronic middle cerebral artery stenosis and the alterations of hemodynamics after PTAS. MATERIAL AND METHODS: In this prospective study, 89 patients with unilateral middle cerebral artery (MCA) stenosis/occlusion were enrolled and classified into four groups according to the degree of stenosis. Cerebral hemodynamics was evaluated by measuring cerebral blood flow (CBF), cerebral blood volume (CBV), and time to peak (TTP) in the ipsilateral and contralateral hemispheres by PCT before and after intervention with PTAS. Differences in hemodynamic parameters before and after intervention were analyzed. RESULTS: Three different hemodynamic patterns were observed in these patients. Patients with severe MCA stenosis (70-99%) or MCA occlusion demonstrated a significant increase of ipsilateral CBV and TTP, indicating hemodynamic compromise. Ten severe stenosis patients with recurrent ischemic symptoms despite of maximal conservative therapy were selected for PTAS. PTAS induced a rapid recovery of cerebral hemodynamics (especially TTP) at 1 week post intervention. CONCLUSION: PCT appears to be a valuable noninvasive technique to evaluate hemodynamic compromise in unilateral chronic MCA stenosis and the improvements after PTAS.

Diagnosis and treatment of autoimmune pancreatitis: experience with 100 patients.

BACKGROUND: Autoimmune pancreatitis (AIP) is increasingly recognized as a unique subtype of pancreatitis. This study aimed to analyze the diagnosis and treatment of AIP patients from a tertiary care center in China. METHODS: One hundred patients with AIP who had been treated from January 2005 to December 2012 in our hospital were enrolled in this study. We retrospectively reviewed the data of clinical manifestations, laboratory tests, imaging examinations, pathological examinations, treatment and outcomes of the patients. RESULTS: The median age of the patients at onset was 57 years (range 23-82) with a male to female ratio of 8.1:1. The common manifestations of the patients included obstructive jaundice (49 patients, 49.0%), abdominal pain (30, 30.0%), and acute pancreatitis (11, 11.0%). Biliary involvement was one of the most extrapancreatic manifestations (64, 64.0%). Fifty-six (56.0%) and 43 (43.0%) patients were classified into focal-type and diffuse-type respectively according to the imaging examinations. The levels of serum IgG and IgG4 were elevated in 69.4% (43/62) and 92.0% (69/75) patients. Pathological analysis of specimens from 27 patients supported the diagnosis of lymphoplasmacytic sclerosing pancreatitis, and marked (>10 cells/HPF) IgG4 positive cells were found in 20 (74.1%) patients. Steroid treatment and surgery as the main initial treatments were given to 41 (41.0%) and 28 (28.0%) patients, respectively. The remission rate after the initial treatment was 85.0%. Steroid was given as the treatment after relapse in most of the patients and the total remission rate at the end of follow-up was 96.0%. CONCLUSIONS: Clinical manifestations, laboratory tests, imaging and pathology examinations in combination could increase the diagnostic accuracy of AIP. Steroid treatment with an initial dose of 30 or 40 mg prednisone is effective and safe in most patients with AIP.

Infective severe acute pancreatitis: a comparison of 99mTc-ciprofloxacin scintigraphy and computed tomography.

AIM: To evaluate (99m)Tc-ciprofloxacin scintigraphy compared with computed tomography (CT) for detecting secondary infections associated with severe acute pancreatitis (SAP) in swine. METHODS: Six healthy swine were assigned to a normal control group (group A, n = 6). SAP was induced in group B (n = 9) and C (n = 18), followed by inoculation of the resulting pancreatic necroses with inactive Escherichia coli (E. coli) (group B) and active E. coli (group C), respectively. At 7 d after inoculation, a CT scan and a series of analyses using infecton imaging (at 0.5, 1, 2, 3, 4 and 6 h after the administration of 370 MBq of intravenous infecton) were performed. The scintigrams were visually evaluated and semi-quantitatively analyzed using region of interest assignments. The differences in infecton uptake and changes in the lesion-background radioactive count ratios (L/B) in the 3 groups were recorded and compared. After imaging detection, histopathology and bacterial examinations were performed, and infected SAP was regarded as positive. The imaging findings were compared with histopathological and bacteriological results. RESULTS: In group A, 6 animals survived without infection in the pancreas. In group B, 7/9 swine survived and one suffered from infection. In group C, 15/18 animals survived with infection. Hence, the number of normal, non-infected and infected SAP swine was 6, 6 and 16, respectively. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the infecton method were 93.8% (15/16), 91.7% (11/12), 92.9% (26/28), 93.8% (15/16) and 91.7% (11/12), whereas these values for CT were 12.5% (2/16), 100.0% (12/12), 50.0% (14/28), 100.0% (2/2) and 46.2% (12/26), respectively. The changes in L/B for the infected SAP were significantly different from those of the non-infected and normal swine (P < 0.001). The mean L/B of the infectious foci at 0.5, 1, 2, 3, 4 and 6 h was 1.17 ± 0.10, 1.71 ± 0.30, 2.46 ± 0.45, 3.36 ± 0.33, 2.04 ± 0.37 and 1.1988 ± 0.09, respectively. At 3 h, the radioactive counts (2350.25 ± 602.35 k) and the mean L/B of the infectious foci were significantly higher than that at 0.5 h (P = 0.000), 1 h (P = 0.000), 2 h (P = 0.04), 4 h (P = 0.000) and 6 h (P = 0.000). CONCLUSION: (99m)Tc-ciprofloxacin scintigraphy may be an effective procedure for detecting SAP secondary infections with higher sensitivity and accuracy than CT.

Focal autoimmune pancreatitis: radiological characteristics help to distinguish from pancreatic cancer.

AIM: To identify the radiological characteristics of focal autoimmune pancreatitis (f-AIP) useful for differentiation from pancreatic cancer (PC). METHODS: Magnetic resonance imaging (MRI) and triple-phase computed tomography (CT) scans of 79 patients (19 with f-AIP, 30 with PC, and 30 with a normal pancreas) were evaluated retrospectively. A radiologist measured the CT attenuation of the pancreatic parenchyma, the f-AIP and PC lesions in triple phases. The mean CT attenuation values of the f-AIP lesions were compared with those of PC, and the mean CT attenuation values of pancreatic parenchyma in the three groups were compared. The diagnostic performance of CT attenuation changes from arterial phase to hepatic phase in the differentiation between f-AIP and PC was evaluated using receiver operating characteristic (ROC) curve analysis. We also investigated the incidence of previously reported radiological findings for differentiation between f-AIP and PC. RESULTS: The mean CT attenuation values of f-AIP lesions in enhanced phases were significantly higher than those of PC (arterial phase: 60 ± 7 vs 48 ± 10, P < 0.05; pancreatic phase: 85 ± 6 vs 63 ± 15, P < 0.05; hepatic phase: 95 ± 7 vs 63 ± 13, P < 0.05). The mean CT attenuation values of f-AIP lesions were significantly lower those of uninvolved pancreas and normal pancreas in the arterial and pancreatic phase of CT (P < 0.001, P < 0.001), with no significant difference at the hepatic phase or unenhanced scanning (P = 0.4, P = 0.1). When the attenuation value increase was equal or more than 28 HU this was considered diagnostic for f-AIP, and a sensitivity of 87.5%, specificity of 100% and an area under the ROC curve of 0.974 (95%CI: 0.928-1.021) were achieved. Five findings were more frequently observed in f-AIP patients: (1) sausage-shaped enlargement; (2) delayed homogeneous enhancement; (3) hypoattenuating capsule-like rim; (4) irregular narrowing of the main pancreatic duct (MPD) and/or stricture of the common bile duct (CBD); and (5) MPD upstream dilation ≤ 5 mm. CONCLUSION: Analysis of a combination of CT and MRI findings could improve the diagnostic accuracy of differentiating f-AIP from PC.

Multi-modality imaging findings of splenic hamartoma: a report of nine cases and review of the literature.

PURPOSE: To investigate the presentation of splenic hamartomas (SHs) on ultrasonography (US), CT and MRI. METHODS: Nine patients (5 males and 4 females, mean age, 52.8 years) with pathologically proven SHs were included in this study. US, CT and MRI images were analyzed retrospectively, and imaging features were correlated with pathological findings. RESULTS: SHs appeared solitary lesion (n = 8) and multiple lesions (n = 1) in the present study. (1) In 8 cases of solitary lesion, the lesions appeared as solid nodules or masses with well-defined margins and varying echogenicity (hyperecho = 5, hypoecho = 2, strong echo = 1) on ultrasound. The lesions showed iso-attenuation (n = 3) or slightly hypo-attenuation (n = 4) on unenhanced CT, and calcification were revealed in 3 lesions. MRI showed isointensity (n = 3) or hypointensity (n = 2) on the T1-weighted image, and heterogeneous hypointensity (n = 2), slightly hyperintensity (n = 2) and hyperintensity (n = 1) on the T2-weighted image. The enhanced patterns of SHs showed mild diffuse heterogeneous enhancement (n = 6) and prominent enhancement (n = 1) during arterial phase and above 7 lesions were demonstrated progressive enhancement at delayed phase on enhanced CT. One lesion without any enhancement was revealed in another patient. (2) One case of multiple lesions included 1 cystic lesion with irregular calcification and 7 solid lesions with progressive enhancement on CT images. CONCLUSIONS: Combination of a variety of imaging modalities could more fully reflect the pathological characteristics and contribute to the diagnosis of SH.

Quantitative perfusion computed tomography measurements of cerebral hemodynamics: correlation with digital subtraction angiography identified primary and secondary cerebral collaterals in internal carotid artery occlusive disease.

BACKGROUND: The aim of the present study was to assess hemodynamic variations in symptomatic unilateral internal carotid artery occlusion (ICAO) patients with primary collateral flow via circle of Willis or secondary collateral flow via ophthalmic artery and/or leptomeningeal collaterals. METHODS: Thirty-eight patients with a symptomatic unilateral ICAO were enrolled in the study. Based on digital subtraction angiography (DSA) findings, patients were classified into 2 groups: primary collateral (n = 14) and secondary collateral (n = 24) groups. Collateral flow hemodynamics were investigated with perfusion computed tomography (PCT) by measuring the cerebral blood flow (CBF), cerebral blood volume (CBV) and time to peak (TTP) in the hemispheres ipsilateral and contralateral to ICAO. Based on the measurements, the ipsilateral to contralateral ratio for each parameter was calculated and compared. RESULTS: Irrespective of the collateral patterns, ipsilateral CBF was not significantly different from that of the contralateral hemisphere (P = 0.285); ipsilateral CBV and TTP was significantly increased compared with those of the contralateral hemisphere (P=0.000 and P = 0.000 for CBV and TTP, respectively). Furthermore, patients with secondary collaterals had significantly larger ipsilateral-to-contralateral ratios for both CBV (rCBV, P = 0.0197) and TTP (rTTP, P = 0.000) than those of patients with only primary collaterals. These two groups showed no difference in ipsilateral-to-contralateral ratio for CBF (rCBF, P = 0.312). CONCLUSION: Patients with symptomatic unilateral ICAO in our study were in an autoregulatory vasodilatation status. Moreover, secondary collaterals in ICAO patients were correlated with ipsilateral CBV and delayed TTP that suggested severe hemodynamic impairment, presumably increasing the risk of ischemic events.

Multidisciplinary team meeting before therapeutic ERCP: A prospective study with 1,909 cases.

OBJECTIVE: To determine the effect of multidisciplinary team meeting (MDTM) on the success rate and complications of therapeutic endoscopic retrograde cholangiopancreatography (ERCP) for hepato-pancreato-biliary diseases. METHODS: All patients undergoing their first therapeutic ERCP over a 21-month period of time in a tertiary care medical center were included. Generally, patients scheduled for ERCP on Friday, Saturday, Sunday, and Monday were subject to MDTM group, and those on Tuesday, Wednesday, and Thursday were allocated to the control group. For each MDTM case, an MDTM was held on the Tuesday prior to the scheduled ERCP. At the meeting, the cases were discussed by a team consisting of chief physicians, radiologists, endoscopists, anesthetists, and surgeons, and a decision was made on the schedule of ERCP. For control cases, a clinical team of one chief physician and two attending physicians made the decision. RESULTS: From April 2006 to December 2007, 912 and 997 ERCP procedures were allocated to the MDTM and control groups, respectively. There was no significant difference in the baseline characteristics and indications between the two groups. Although the success rates were not significantly different between MDTM and control groups (82.9% vs. 84.8%, P=0.321), MDTM was significantly associated with a decreased overall complication rate of (6.9% vs. 12.0%, p<0.001) and severe complication rate (0.4% vs. 2.5%, p=0.035). CONCLUSIONS: Pre-ERCP MDTM decreases the frequency and severity of ERCP-related complications, with similar success rate, compared to routine practice.

CT features of colloid carcinomas of the pancreas.

BACKGROUND: Colloid carcinomas of the pancreas have better prognosis than ordinary ductal adenocarcinoma, and preoperative distinction of colloid carcinoma from other pancreatic tumors is valuable for patient therapeutic planning and prognosis assessment. However, data about CT features of colloid carcinoma are very limited. This study aimed to investigate the CT features of this tumor. METHODS: Institutional review board approval was obtained for this study. Seven patients with pathologically proven colloid carcinoma of the pancreas were included. Unenhanced and dynamic enhanced CT was performed in all the patients. CT features were analyzed retrospectively and correlations with pathological findings were evaluated. RESULTS: Mean age of the patients was 59.8 years (41 - 76 years). Five tumors were located in the pancreatic head, and the other two in body and tail respectively. The maximum mean diameter of the tumors on axial scanning was 3.9 cm (3.0 - 6.7 cm). Tumors were round (n = 5) and lobular (n = 2). Tumors appeared slight hyp-attenuation on unenhanced CT, and peripheral and internal meshlike progressive delayed enhancement with great percent of cystic areas on enhanced CT. Calcification and gas in the tumor was seen in one patient whose duodenum was invaded by the tumor. CONCLUSIONS: Colloid carcinomas of the pancreas appear as round or labular masses with great percent of cystic areas and slight hyp-attenuation on unenhanced CT and peripheral and internal meshlike progressive delayed enhancement on enhanced CT.

[In vivo proton magnetic resonance spectroscopy of hepatocellular carcinoma and cholangiocarcinoma].

OBJECTIVE: To investigate the value of in vivo proton magnetic resonance spectroscopy (1H MRS) in the assessment of hepatocellular carcinoma (HCC) and cholangiocarcinoma. METHODS: 1H MRS was performed in normal volunteers and in patients with pathologically confirmed HCC and cholangiocarcinomas using a whole-body 1.5-T scanner. The choline-to-lipid ratios were measured by dividing the peak area of choline at 3.2 ppm and lipid at 1.3 ppm. RESULTS: The ratio of choline-to-lipid for normal liver, cholangiocarcinomas, and HCC were 0.07 +/- 0.04, 0.11 +/- 0.06, and 0.52 +/- 0.15, respectively. The ratio of choline-to-lipid was significantly higher in HCC compared than those in cholangiocarcinomas or normal livers (P < 0.05). However, it was not significantly different between cholangiocarcinomas and normal livers (P > 0.05). CONCLUSION: In vivo 1H MRS can reflect the pathological changes of HCC and cholangiocarcinomas at metabolic level and thus is useful in the diagnosis of these two cancers.

Telomerase activity (TMA) in tumour and peritumoural tissues in a rat liver cancer model.

PURPOSE: To study the levels of telomerase activity (TMA) in tumour and peritumoural tissues in a liver cancer model in rats, and to study the change in TMA expression over time. METHODS: Using the telomeric repeated amplification protocol (TRAP), TMA was measured in tumour tissue, peritumoural tissue and normal liver tissue of Walker-256 tumour-bearing rats at 4, 6 and 8 days after tumour implantation. RESULTS: TMA at day 4, 6 and 8 was 0.767+/-0.117, 0.768+/-0.118 and 0.774+/-0.111 in tumour tissue, 0.389+/-0.263, 0.492+/-0.253 and 0.584+/-0.239 in peritumoural tissue, and 0.231+/-0.022, 0.229+/-0.022 and 0.233+/-0.021 in normal liver tissue, respectively. TMA in tumour tissue was higher than that in peri-tumour and normal liver tissues at all time points of measurement (P < 0.05). The TMA levels in tumour tissue and normal liver tissue did not show any change over time. TMA level in the peritumoural tissue increased with time; TMA level in animals sacrificed at day 8 was higher than that seen in animals sacrificed at day 4 (P < 0.05). CONCLUSION: TMA in walker-256 tumour-bearing rats was higher than that in normal and peritumoural tissues. TMA level in the peritumoural tissue increased with time suggesting that TMA activation in peritumoural tissue may be an important factor promoting tumour growth.

CT-guided percutaneous ethanol injection with disposable curved needle for treatment of malignant liver neoplasms and their metastases in retroperitoneal lymph nodes.

AIM: To explore the feasibility of computed tomography (CT)-guided percutaneous ethanol injection (PEI) using a disposable curved needle for treatment of malignant liver neoplasms and their metastases in retroperitoneal lymph nodes. METHODS: CT-guided PEI was conducted using a disposable curved needle in 26 malignant liver tumors smaller than 5 cm in diameter and 5 lymph node metastases of liver cancer in the retroperitoneal space. The disposable curved needle was composed of a straight trocar (21G) and stylet, a disposable curved tip (25 G) and a fine stylet. For the tumors found in deep sites and difficult to reach, or for hepatic masses inaccessible to the injection using a straight needle because of portal vein and bile ducts, the straight trocar was used at first to reach the side of the tumor. Then, the disposable curved needle was used via the trocar. When the needle reached the tumor center, appropriate amount of ethanol was injected. For relatively large malignant liver tumors, multi-point injection was carried out for a better distribution of the ethanol injected throughout the masses. The curved needle was also used for treatment of the metastasis in retroperitoneal lymph nodes blocked by blood vessels and inaccessible by the straight needle. RESULTS: All of the 26 liver tumors received 2 or more times of successful PEI, through which ethanol was distributed throughout the whole tumor mass. Effect of the treatment was monitored by contrast-enhanced multi-phase CT and magnetic resonance imaging (MRI) examinations three months later. Of the 18 lesions whose diameters were smaller than 3 cm, the necrotic change across the whole mass and that in most areas were observed in 15 and 3 tumors, respectively. Among the 8 tumors sizing up to 5 cm, 5 were completely necrotic and 3 largely necrotic. Levels of tumor seromarkers were significantly reduced in some of the cases. In 5 patients with metastases of liver cancer in retroperitoneal lymph nodes who received 1 to 3 times of PEI, all the foci treated were completely necrotic and smaller demonstrated by dynamic contrast-enhanced CT or MRI 3 months later. CONCLUSION: CT-guided PEI using a disposable curved needle is effective, time-saving and convenient, providing an alternative therapy for the treatment of malignant liver tumors and their retroperitoneal lymph node metastases.