Histopathological and immunohistochemical clues to the illusive diagnosis of follicular dendritic cell sarcoma: A clinicopathological masquerader

Follicular dendritic cell sarcoma is a rare histiocytic and dendritic neoplasm mainly involving the lymph nodes and selective extranodal sites. They are often misdiagnosed due to nonspecific clinical, radiological, and morphological findings in addition to their rarity. Four cases described below had variable age of presentation, site of involvement, size of the lesion, and histopathological features. Application of an extensive immunohistochemical panel, including a combination of >1 dendritic cell marker, clinched the diagnosis. A combination of D2-40 and Cluster of differentiation 21 (CD21) worked best in establishing a definite role in the current series. Programmed death-ligand 1 (PD-L1) analysis was positive in two of the three cases where it could be performed. However, none of our cases had received immunotherapy. Prompt recognition of the described histopathology features and incorporation of novel immunohistochemical markers can translate to timely initiation of therapy for this aggressive disease

Application of D2-40/CD34-CK cocktail antibodies for colorectal cancer with insufficient lymph node harvest / 临床与实验病理学杂志

Purpose To investigate the value of application of D2-40/CD34-CK cocktail antibodies by double immunohistochemical staining for assessment of lymphovascular invasion (LVI) and to determine its prognostic significance in colorectal cancer with insufficient lymph node harvest. Methods Specimens from 133 cases of colorectal cancer with less than 12 lymph nodes were selected. HE staining and double immunohistochemical staining of the cocktail antibodies were performed to compare the difference of the two methods in screening for LVI. The The relationship between LVI confirmed by cocktail antibody immunohistochemical staining and clinicopathological characteristics and overall survival (OS) of patients was analyzed. Results (1) The detection rates of cocktail antibody double immunohistochemical staining and HE staining for LVI were 42.9％ (57/133) and 21.8％ (29/133) with statistically significant difference (P < 0.001). (2) The presence of LVI confirmed by double staining was significantly associated with Dukes staging, depth of invasion, clinical stages, lymph node metastasis and tumor budding (P < 0.05). (3) The presence of LVI, the location and extent of LVI, and the number of tumor cells in thrombus ≥5.5 for cases with LVI ≤2 clusters, were significantly associated with OS (P < 0.05). Conclusion D2-40/CD34-CK cocktail antibodies double staining is superior to routine HE staining in assessing LVI. LVI is intimately associated with tumor stage, lymph nodes metastasis and tumor budding, and it is an independent prognostic factor for CRC patients. It should be a supplementary examination for these patients with insufficient lymph node harvest.

Difference in expression of lymphatic markers D2-40 and Prox1 between angiokeratoma corporis diffusum and angioma serpiginosum / 中华皮肤科杂志

Objective@#To investigate the significance of lymphatic markers in the differential diagnosis of angiokeratoma corporis diffusum (ACD) and angioma serpiginosum (AS) .@*Methods@#Totally, 9 patients with ACD and 6 with AS were enrolled from Department of Dermatology, Xijing Hospital between 2006 and 2017, and their clinical and histopathological features were retrospectively analyzed. Skin sections from all the patients were stained for CD31, D2-40 and Prox1.@*Results@#In the 9 patients with ACD, abnormal vessels were weakly positive or negative for CD31, and positive for Prox1. Endothelial cells in abnormal vessels were locally positive for D2-40 in 4 patients with ACD, but negative for D2-40 in the other 5 patients. In the 6 patients with AS, the endothelia of hyperplastic small vessels were positive for CD31, but negative for D2-40 and Prox1.@*Conclusion@#The clinical features of a few patients with ACD are similar to those of patients with AS, and lymphatic markers have definite significance in the differential diagnosis of the two diseases.

Comparison of lymphangiogenesis, lymphatic invasion, and axillary lymph node metastasis in breast carcinoma

Context: Lymphangiogenesis correlates with poor prognosis in Invasive Ductal Carcinoma (IDC) breast. D2-40 antibody, a specific marker for lymphatic endothelium, differentiates lymphatic from vascular endothelium. Therefore, the aims of this study were to estimate lymphangiogenesis using D2-40 antibody and correlate with lymphatic invasion (LI) and axillary lymph node (LN) status and compare lymphatic mean vessel density (LMVD) with Tumor (T) and Node (N) stages and grade of tumor. Methods and Material: The study was conducted on fifty consecutive cases of IDC breast who underwent modified radical mastectomy (MRM) from Jan 2009 to March 2011. Hematoxylin-eosin sections and Immunohistochemistry (IHC) slides were studied along with their LN status. LMVD was counted after D2-40 immunostaining (100x magnification) in three hot spots in peritumoral areas and averaged. LI as opposed to vascular invasion (BVI), and LN status for all cases were assessed. Statistical Analysis: Statistical analysis was done using SPSS software (version 14.0 for Windows). Pearson's correlations, χ2 tests and Mann-Whitney U test were used. Results: Lymphangiogenesis varied from 0 to 58 with mean LMVD of 11. Of 50 cases, five showed no lymphatic vessels in peritumoral areas; of these five, three had positive LNs. 21/50 cases had LI. No statistical significant association was seen between lymphangiogenesis and LI. 34/50 cases had positive LNs. Mean LMVD was higher in patients with N2/N3 stage as compared to N0/N1 stage and was statistically significant (P = 0.013). Conclusions: D2-40 is specific marker for lymphatic endothelium. LI and lymphangiogenesis, as opposed to BVI, are better prognostic indicators in IDC breast.

Immunohistochemical evaluation of lymphovascular invasion in carcinoma breast with CD34 and D2-40 and its correlation with other prognostic markers

Background: Carcinoma breast is ever-evolving and becoming increasingly prevalent in India. Numerous prognostic factors based on morphology and immunohistochemistry (IHC) have been established which need to be interconnected to give patients best possible treatment. Aims: This study aims to confirm and analyze lymphovascular invasion (LVI) detected by hematoxylin and eosin (H and E) using IHC with CD34 and D2-40 and its correlation with other biologic and morphologic prognostic markers. Settings and Design: This was a prospective study. Materials and Methods: Fifty mastectomy specimens diagnosed as infiltrating ductal carcinoma breast on histopathology selected for the study. Evaluation of formalin-fixed paraffin-embedded sections was done using H and E and IHC for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 HER2/neu receptors, CD34, and D2-40 endothelial markers. Correlation of LVI done with prognostic markers of Carcinoma Breast, namely, age of the patient, tumor size, Nottingham grade, lymph node ratio (LNR), Nottingham prognostic index (NPI), ER/PR status, and HER2/neu status. CD34 and D2-40 utilized to distinguish blood vessel, lymph vessel, and retraction artifacts and to calculate lymphatic microvessel density (LMVD) and blood microvessel density (BMVD). Statistical Analysis Used: SPSS Software Package. Results: LVI was associated with younger age (P = 0.001), greater tumor size (P = 0.007), higher Nottingham grade (P = 0.001), higher LNR (P = 0.001), higher NPI (P = 0.001), Negative ER Status (P = 0.001), Negative PR Status (P = 0.002), Positive HER2/neu status (P = 0.021), Higher Intratumoral BMVD (P = 0.016), Peritumoral BMVD (P = 0.001), and Intratumoral LMVD (P = 0.009). Blood vessels more commonly invaded than lymph vessels. Retraction artifacts can be mistaken for LVI without IHC. Conclusions: D2-40 is a promising marker for lymphatic endothelium. LVI is a poor prognostic marker hence should be evaluated imperatively in all cases of carcinoma breast.

A Case of Isolated Conjunctival Lymphangioma Mimicking a Recurrent Conjunctival Cyst

PURPOSE: To report a case of isolated conjunctival lymphangioma mimicking a recurrent conjunctival cyst. CASE SUMMARY: A 39-year-old male with a conjunctival cyst in the right eye lasted for 1 month visited our hospital. He had previously undergone aspiration of the cyst at another hospital 1 week before visiting our hospital. However, the cyst recurred, and he was referred to our hospital. On slit lamp biomicroscopy, yellow-colored turbid fluid and a hemorrhage were observed in the conjunctival cyst, but no specific finding was found in the fundus photography. The patient was initially treated with topical antibiotics and steroids. Three weeks later, absorption of the hemorrhage was noted, but there was no change in the size of the cyst. Therefore, surgical removal and histological examination of the cyst were performed. The histological examination revealed that the lesion was positive for CD 31 and D2-40, and the cyst was diagnosed as a cystic conjunctival lymphangioma. Thereafter, brain magnetic resonance imaging was performed to screen for orbital lymphangioma and systemic disease that could accompany a conjunctival lymphangioma. However, no specific findings were observed. There was no recurrence of the conjunctival cyst at 1 year and 6 months after surgical removal, and no other ophthalmic or systemic complication was observed. CONCLUSIONS: An isolated conjunctival lymphangioma is a rare lesion. In the diagnosis of lymphangioma, systemic examination is recommended for the identification of comorbid diseases, such as orbital lymphangioma.

Mechanism of the "negative development" of carbon nanoparticles for the parathy-roid gland in thyroidectomy through expression contrast of vascular-related markers / 中国肿瘤临床

Objective: To investigate the anatomical characteristics of the parathyroid lymphatic system and the mechanism of the"negative development"of the carbon nanoparticles for parathyroid gland in thyroidectomy.Methods:This retrospective study used parathyroid tissue samples from patients that were obtained from archival records in the pathology department,including 45 cases of normal parathyroid gland tissues that were accidentally resected in thyroidectomy,10 cases of parathyroid adenomas,and 7 cases of parathyroid carcinoma.Ten cases of normal thyroid tissues were selected as positive control.Immunohistochemistry was performed using the antibodies specific for lymphatic endothelium,such as D2-40 and LYVE-1,and antibodies specific for vascular endothelial cell such as CD31 and CD34,to distinguish them from each other.Results:A total of 62 parathyroid glands samples were stained with vas-cular markers CD31,CD34 and lymphatic markers D2-40,LYVE-1 respectively(partial samples were stained unsuccessfully).Vascular vessels in the CD31 staining group were detected in 50 of 58 examined glands and the positive rate was 86.2%.In the CD34 staining group,positive rate was 100%(60/60).The positive cells were found in the central,periphery and vascular hilum of the glands.Howev-er,lymph vessels in the D2-40 staining group were detected from 17 out of 59 examined glands,with the positive rate of 28.8%;In the LYVE-1 staining group,positive rate was 39.6%(23/58).The positive cells were found in the membrane or vascular hilum,less frequent or undetectable in the central portion.Conclusions:Most of the parathyroid glands of adults might lack a lymphatic network.Only a few adult parathyroid glands had minority lymph vessels,and these lymphatics generally localized at the membrane area or in the vas-cular hilum, which could be one of the main and anatomical mechanisms resulting in drainage failure or obstruction of carbon nanoparticles and thus in parathyroid"negative development."

Clinical application of cocktail double enzyme labeled antibody(D2-40/CK19)staining for the diagnosis of lymphatic vessel invasion in papillary thyroid carcinoma / 中华内分泌代谢杂志

Objective To analyse the relationship between lymphatic vessels invasion and clinical pathological features of papillary thyroid carcinoma ( PTC ) .Methods The expressions of D2-40 and CK19 were examined in the 104 specimens of PTC using immunohistochemical staining with combined monoclonal antibodies and cocktail double enzyme labeled antibody( D2-40/CK19) stainings.The two methods were compared in the diagnosis of PTC metastasis, and the factors affecting lymphatic vessels formation were analyzed.Results The positive rate of lymphatic vessels invasion was 37.5%(39/104) by using immunohistochemical staining with combined monoclonal antibodies and 53.8%( 56/104 ) by cocktail double enzyme labeled antibody ( D2-40/CK19 ) staining ( P<0.05).The lymph node metastasis rate was 83.9%(47/56) in the group with lymphatic vessels invasion, significantly higher than that without invasion 22.9%(11/48, P<0.01).The age of patients, diameter of primary tumor were the influence factors of lymphatic vessels invasion in PTC patients(P<0.05 and P=0.063).Conclusion Cocktail double enzyme labeled antibody ( D2-40/CK19 ) staining is a better method to detect lymphatic vessels invasion in PTC than immunohistochemical staining with combined monoclonal antibodies.

Sebaceoma：a clinicopathological analysis of 31 cases / 中华皮肤科杂志

Objective To analyze clinicopathologic features of sebaceoma. Methods Clinical, pathologic and immunohistochemical findings from 31 cases of sebaceoma were retrospectively analyzed. The clinicopathologic features of sebaceoma were investigated. Results There were 9 males and 22 females. The patients′ age was 53.90 ± 15.40 years, and the clinical course was 9.41 ± 13.75 years. Sebaceoma predominantly affected the face. The common lesion of sebaceoma was red, yellowish?red, skin?colored or slight brown papules, with no subjective symptoms in most cases. Histopathologically, neoplasms had symmetric structures, and were located in the dermis. Epidermal involvements were found in 9 cases. The neoplasm cells were mainly composed of basaloid cells, a few mature sebocytes and some transition cells. The proportion of mature sebocyts was less than 1%in 26 cases, less than 20%in 2 cases, and 20%-40%in 3 cases. Mitoses were occasionally found in 5 cases. One patient was complicated by eccrine poroma. Varying amounts of ducts were found in all the patients. Immunohistochemical staining showed that epithelial membrane antigen was expressed on ducts and mature sebocytes in all the patients, while epithelial antigen was undetected in any of the patients. Carcinoembryonic antigen, androgen receptor and D2?40 were found in 20, 24 and 28 patients with sebaceoma, respectively. Conclusions The diagnosis of sebaceoma mainly depends on histopathological examination. Combined immunohistochemical detection of epithelial membrane antigen, androgen receptor and D2?40 is beneficial to its differential diagnosis.

Immunohistochemical Expressions of D2-40, CD34, and Factor XIIIa between Dermatofibroma and Dermatofibrosarcoma Protuberance / 대한피부과학회지

BACKGROUND: Dermatofibrosarcoma protuberance (DFSP) must be differentiated from dermatofibroma (DF). However, especially in cases of superficial biopsy and cellular dermatofibroma, this is difficult by using histopathology alone since both are composed of neoplastic spindle cells. Although a panel of immunostains is useful, the expressions of conventional markers often overlap. A previous study showed that novel D2-40 immunostain may be useful for differentiating between DF and DFSP. OBJECTIVE: To evaluate the usefulness of D2-40 immunohistochemical staining for differentiating DFSP from DF and compare the results with other commonly used immunostains (CD34 and factor XIIIa). METHODS: Twenty-eight cases of DF and 15 cases of DFSP were selected from clinicopathologically proven cases reviewed by the Department of Dermatology at our medical center and Daegu Catholic University Medical Center. D2-40, CD34, and factor XIIIa immunohistochemical staining was performed. The immunopositivity was measured throughout the entire lesion. RESULTS: Seventeen cases (60.7%) of DF and no cases of DFSP showed immunoreactivity to D2-40 in the spindle cells. Three (10.7%) cases of DF and 13 (86.7%) cases of DFSP showed immunoreactivity to CD34 in the spindle cells. Twenty-five (89.3%) cases of DF and four (26.7%) cases of DFSP showed immunoreactivity to factor XIIIa in the spindle cells. A total of 60.7% of cases of DF were positive on D2-40 staining, 89.3% were negative on CD34 staining, and 89.3% were positive on factor XIIIa staining. All cases (100%) of DFSP were negative by D2-40 staining, 86.7% were positive by CD34 staining, and 73.3% were negative by factor XIIIa staining. CONCLUSION: D2-40 immunostaining may be useful for distinguishing between DF and DFSP since the immunoreactivity of DF was significantly higher than that of DFSP (p=0.001). However, the results of our study were not as useful as those of a previous study. Therefore, further studies are needed to address this issue.

Role of podoplanin in potentially malignant disorders and oral squamous cell carcinoma and its correlation with lymphangiogenesis.

BACKGROUND: Oral squamous cell carcinoma (OSCC) ranks as one of the most common types of cancer and oral potentially malignant lesions (OPMLs) provides with an overall increased risk for development of carcinoma. As podoplanin expression is attracting interest as a marker for cancer diagnosis and prognosis, this study assesses the role of podoplanin expression in such lesions. MATERIALS AND METHODS: Podoplanin expression and lymphatic vessel density (LVD) was determined using D2‑40, a marker for podoplanin, in 70 diagnosed cases of potentially malignant lesions and OSCC. RESULTS: Normal epithelium showed negligible podoplanin expression, whereas the expression extended predominantly at the basal layer and the suprabasal layer or above at one or multiple areas in potentially malignant lesions. Podoplanin expression in OSCC showed two different patterns‑diffuse and focal. A statistically significant increase in mean LVD was seen from normal epithelium to potentially malignant lesions (P < 0.001) and to OSCC (P < 0.022) while a non‑significant increase was seen (P < 0.594) between OPMLs and OSCC. Overall no significant correlation was found between D2‑40 epithelial positivity and LVD (P = 0.122). CONCLUSION: This study suggests the utility of podoplanin as a biomarker for cancer risk assessment as it detects the early changes and thus provides an additional value beyond current clinical and histopathological evaluations. Hence, podoplanin is suggested to be a marker of tumor initiation and to a lesser extent of tumor progression.

Diagnostic utility of immunohistochemical markers SALL4, D2-40 and Glypican-3 in primary testicular germ cell tumors / 临床与实验病理学杂志

Purpose To investigate the diagnostic utility of the immunohistochemical markers SALL4, D2-40 and Glypican-3 in prima-ry testicular germ cell tumors (TGCTs). Methods The expression of SALL4, D2-40 and Glypican-3 protein was detected by EnVi-sion immunohistochemical method in 56 cases of primary testicular germ cell tumors, including 5 intratubular germ cell neoplasms ( IT-GCNs) , 10 seminomas, 14 embryonal carcinomas ( ECs) , 14 yolk sac tumors ( YSTs) , 1 choriocarcinoma, 5 immature teratomas and 12 mature teratomas. 10 normal testicular tissues and 5 lymphomas were selected as control. Results All of ITGCNs, seminomas, YSTs and ECs were diffusely strongly positive for SALL4. Focal SALL4 staining was seen in choriocarcinoma, 3 of 5 immature terato-mas and 3 of 12 mature teratomas. All of ITGCNs, seminomas showed diffusely strong D2-40 staining. ECs (4/14) were focally posi-tive for D2-40, while choriocarcinoma, YSTs and teratomas were negative for D2-40. Glypican-3 was diffusely positive in YSTs (13/14), and focally weakly positive in ECs (2/14), respectively. ITGCNs, seminomas, choriocarcinoma and teratoma were negative for Glypican-3. In contrast, 10 normal testicular tissues and 5 lymphomas showed no SALL4, D2-40 and Glypican-3 staining. Conclu-sions SALL4 is a useful diagnostic marker with high sensitivity and specificity for TGCTs. Combination of SALL4, D2-40 and Glypi-can-3 is helpful to the diagnosis and differential diagnosis for TGCTs.

Clinicopathological significance of D2-40 expression in craniopharyngioma / 临床与实验病理学杂志

Purpose To investigate the expression pattern and c1inicopatho1ogica1 significance of D2-40 in craniopharyngioma( CP). Methods Immunohistochemica1 method was used to assess D2-40 expression in 126 cases of craniopharyngioma. Statistic software was used to ana1yze the corre1ation between D2-40 expression and c1inicopatho1ogica1 features. Results The overa11 positive rate of D2-40 expression in 126 craniopharyngioma was 87. 30%. The rate of +, and  were 44. 44%( 56/126 ),37. 30%( 47/126 )and 5. 56%(7/126)respective1y. In adamantinomatous CPs,D2-40 expression was observed in epithe1ia1 components corresponding to the stratum intermedium,whi1e in papi11ary CPs,it was immuno-positive in basa1 ce11s. With the increasing of the existence of inf1amma-tion in tumor,D2-40 expression was up-regu1ated. Tumor ce11s were over-expressed for D2-40 in basa1 ce11s and stratum intermedium ce11s of the invasive frontier. D2-40 expression was higher in the invasive craniopharyngioma than in the non-invasive craniopharyngio-ma. In addition,D2-40 expression was higher in the recurrent craniopharyngioma than in the non-recurrent craniopharyngioma. There were no significant re1ationship between D2-40 expression and gender,and histo1ogica1 c1assification of tumors. Conclusions D2-40 expression is associated with the deve1opment of craniopharyngioma,which can 1ead to enhance the tumor ce11 invasion,and may be re-1ated to inf1ammation-re1ated mechanisms. D2-40 expression may be one of the recurrence factors in patients with craniopharyngioma.

Expression of VEGF-D, D2-40, and MMP-7 and its relationship with cervical lymph node metastasis in thyroid papillary carcinoma / 中华内分泌外科杂志

Objective To detect the expression levels of vascular endothelial growth factor D ( VEGF-D) , D2-40 and MMP-7 in papillary thyroid carcinoma ( PTC) and to explore their relationship with micro lymphat-ic density, distribution,and cervical lymph node metastasis .Methods SP immunohistochemistry was used to study the expression of VEGF-D, D2-40 and MMP-7 in 38 cases of PTC with cervical lymph node metastasis , 45 cases of PTC without cervical lymph node metastasis , and 22 cases of normal thyroid tissues .Micro lymphatic density ( MLD) was counted .Results The expression level of VEGF-D in PTC with cervical lymph node metasta-sis and normal thyroid tissues had significant difference (χ2 =13.074, P=0.000, P<0.0167).The expression level of D2-40 in PTC with and without cervical lymph node metastasis had significant difference ( P<0.05 ) . MMP-7 expression in PTC with and without cervical lymph node metastasis had statistical significance ( 91.67%vs 73.33%, P<0.05).MLD in PTC with and without cervical lymph node metastasis and in normal thyroid tis-sues had statistical difference (11.7 ±3.5 vs 8.9 ±3.1 vs 3.9 ±2.7, P<0.05).Conclusion VEGF-D, D2-40 and MMP-7 are highly expressed in PTC and they are possibly related with cervical lymph node metastasis of PTC.

Application of D2-40/S100 and CD34/S100 detection in the diagnosis of blood and lymphatic vessel invasion of cutaneous malignant melanoma / 中华皮肤科杂志

Objective To develop an immunohistochemical assay for the diagnosis of cutaneous malignant melanoma (CMM) micrometastasis via blood and lymphatic vessels,and to evaluate its clinical significance.Methods Fifty-three patients (32 males and 21 females) histopathologically diagnosed as CMM were enrolled in this study.The patients were aged (61.2 ± 8.4) years (range,52-72 years).Tissue specimens were obtained from the central area of tumor in each case,and also from removed lymph nodes in some cases.The average duration of follow-up was (65.00 ± 5.68) months.During the follow-up,17 patients died of the recurrence or metastasis of CMM,and 6 patients were lost to follow-up.The expressions of D2-40,S100 and CD34 antigens in 53 tissue specimens were examined by immunohistochemical staining with three individual monoclonal antibodies,or by an immunohistochemical method using 2 two-antibody cocktails (D2-40/S 100 and CD34/S100) and double-color chromogens in single tissue sections.Results Of the 53 patients,30.19％ (16/53) were positive for hematoxylin-eosin (HE) staining combined with immunohistochemical staining with individual monoclonal antibodies,and 49.06％ (26/53) for the immunohistochemical method using two-antibody cocktails and double-color chromogens.Statistical differences were found in the positive rate between the two methods (x2 =3.94,P＜ 0.05).Compared with patients without blood/lymph vessel tumor emboli,those with blood/lymph vessel tumor emboli showed higher lymph node metastasis rate (80.77％ (21/26) vs.37.04％ (10/27),x2 =10.43,P ＜ 0.001),but lower five-year survival rate (42.31％ (11/26) vs.70.37％ (19/27),x2 =4.25,P ＜ 0.05).Conclusions The immunohistochemical method with two-antibody cocktails is superior to HE staining combined with immunohistochemical staining with individual monoclonal antibodies in the detection of blood/lymph vessel tumor emboli.And blood/lymph vessel tumor emboli may be an important prognostic factor in patients with CMM.

Comparison of diagnostic values of Clusterin, CXCL13, Podoplanin, CD21 and CD35 in foll-cular dendritic cell sarcoma / 临床与实验病理学杂志

Purpose To evaluate the diagnostic values of Clusterin, CXCL13, Podoplanin (D2-40), CD21 and CD35 in follcular den-dritic cell sarcoma. Methods The expression levels of 10 cases of follcular dendritic cell sarcoma ( FDCS) and 12 types of FDCS mimics (83 cases in total) were investigated by immunohistochemical methods, the latter including solitary fibrous tumor, leiomyosar-coma, gastrointestinal stromal tumor and others. The diagnostic validities of the five biomarkers were compared. Results ( 1 ) The positive rates of Clusterin, CXCL13, D2-40, CD21 and CD35 in the FDCS group were 100%, 70%, 60%, 90% and 80%, respec-tively, the corresponding rates in the control group were 30%, 4%, 11%, 2% and 0 in turn. (2) The five biomarkers could be cate-gorized into 3 groups, according to their diagnostic values in FDCS. The first group included CD21 and CD35, which had much higher sensitivities (90%, 80%), specificities (100%, 98%) and accuracies (98%, 96%), compared with the other biomarkers. The second group included CXCL13 and D2-40, which had a relatively lower sensitivities (70%, 60%), specificities (96%, 89%) and accuracies (94%, 86%), compared with CD21 and CD35. The third group included Clusterin, which had the highest sensitivity (100%), while the specificity (70%)and accuracy (73%) were inferior to the first and second groups. (3) The diagnostic values of CD21 and CD35 combination were 100%, 98%, 98%, 83% and 100%, respectively. Conclusions (1) CD21 and CD35 are the most valuable biomarkers for FDCS. The combined diagnostic effect of the two markers is generally superior to that of single marker. (2) Clusterin has the highest sensitivity for FDCS. However, the frequent expression in FDCS mimickers restricts its diagnostic values for FDCS. (3) CXCL13 and D2-40 may be used as a marker for FDCS, but are not recommended as the first selection based on their diagnostic performance. (4) On the reasons that FDCS mimickers may not uncommonly express Clusterin and D2-40, and rarely show positivity for CD21 or CXCL13, more attention should be paid to the phenomenon to avoid misdiagnosis.

The role of intratumoral lymphovascular density in distinguishing primary from secondary mucinous ovarian tumors

OBJECTIVE: Ovarian mucinous metastases commonly present as the first sign of the disease and are capable of simulating primary tumors. Our aim was to investigate the role of intratumoral lymphatic vascular density together with other surgical-pathological features in distinguishing primary from secondary mucinous ovarian tumors. METHODS: A total of 124 cases of mucinous tumors in the ovary (63 primary and 61 metastatic) were compared according to their clinicopathological features and immunohistochemical profiles. The intratumoral lymphatic vascular density was quantified by counting the number of vessels stained by the D2-40 antibody. RESULTS: Metastases occurred in older patients and were associated with a higher proportion of tumors smaller than 10.0 cm; bilaterality; extensive necrosis; extraovarian extension; increased expression of cytokeratin 20, CDX2, CA19.9 and MUC2; and decreased expression of cytokeratin 7, CA125 and MUC5AC. The lymphatic vascular density was increased among primary tumors. However, after multivariate analysis, the best predictors of a secondary tumor were a size of 10.0 cm or less, bilaterality and cytokeratin 7 negativity. Lack of MUC2 expression was an important factor excluding metastasis. CONCLUSIONS: The higher intratumoral lymphatic vascular density in primary tumors when compared with secondary lesions suggests differences in the microenvironment. However, considering the differential diagnosis, the best discriminator of a secondary tumor is the combination of tumor size, laterality and the pattern of expression of cytokeratin 7 and MUC2. .

The role of immunohistochemistry in the detection of vascular invasion in specimens of endoscopic submucosal dissection / O papel da imuno-histoquímica na detecção de invasão vascular em espécimes de dissecção endoscópica da submucosa

INTRODUCTION: Endoscopic submucosal dissection (ESD) of early neoplasias of the gastrointestinal tract (GIT) has been increasingly applied as an alternative to invasive surgical procedures, with the aim to preserve the patient's organ and quality of life, although it does not allow the histopathological analysis of lymph nodes. Previous studies demonstrated that the presence of neoplastic emboli in lymphatic (lymphatic vascular invasion [LVI]) or blood vessels (blood vascular invasion [BVI]) is considered a positive predictive factor for the occurrence of lymph node metastasis. The assessment of vascular invasion carried out only by routine hematoxylin and eosin staining (HE) may yield both falsepositive and false-negative results. D2-40 is a specific monoclonal antibody to the lymphatic endothelium. Thus, it is useful for identifying LVI and distinguishing if tumor embolization is found in blood or lymphatic vessels. OBJECTIVE: To determine the role of immunohistochemistry (IHC) in the assessment of ESD specimens by comparing the detection of LVI and BVI by HE and IHC with D2-40 and CD34 immunolabeling. METHOD: We conducted the IHC study using D2-40 and CD34 markers (pan-endothelial) in 30 cases of ESD with histological diagnosis of carcinoma in order to assess the presence of LVI and BVI. RESULTS: The detection of LVI was more prevalent than BVI. Three out of six cases with LVI were false-positive by HE and six were false-negative by IHC. Regarding BVI, five cases were identified and one was false-negative by IHC. CONCLUSION: Our results indicated that the histopathological analysis of ESD specimens by exclusively routine HE staining does not allow proper evaluation of BVI or LVI.

INTRODUÇÃO: A dissecção endoscópica da submucosa (DES) de neoplasias precoces do trato gastrointestinal (TGI) tem sido cada vez mais aplicada como alternativa aos procedimentos cirúrgicos invasivos, visando a preservar o órgão e a qualidade de vida do paciente, contudo, não possibilita a avaliação histopatológica de linfonodos. Estudos anteriores demonstraram que a presença de êmbolos neoplásicos, em vasos linfáticos (invasão vascular linfática [IVL]) ou sanguíneos (invasão vascular sanguínea [IVS]), é considerada um fator preditivo positivo para ocorrência de metástase linfonodal. A avaliação da invasão vascular realizada apenas pela coloração de rotina hematoxilina e eosina (HE) pode gerar resultados falso-positivos e falso-negativos. O D2-40 é um anticorpo monoclonal específico para endotélio linfático, sendo, portanto, útil para identificar IVL e distinguir se a embolização tumoral encontra-se em vasos sanguíneos ou linfáticos. OBJETIVO: Determinar o papel do estudo imuno-histoquímico (IHQ) na avaliação de espécimes de DES, comparando a detecção de IVL e IVS, pelo HE e IHQ com marcação por D2-40 e CD34. MÉTODO: Foi realizado estudo IHQ utilizando os marcadores D2-40 e CD34 (pan-endotelial) em 30 casos de produtos de DES com diagnóstico histológico de carcinoma para avaliar a presença de IVL e IVS. RESULTADOS: A detecção de IVL foi maior que a de IVS. Dos seis casos com IVL ao HE, três eram falso-positivos e seis, falso-negativos à IHQ. Em relação à IVS, foram identificados cinco casos falsopositivos e um falso-negativo à IHQ. CONCLUSÃO: Nossos resultados indicaram que a análise histopatológica dos produtos de DES realizando apenas a coloração HE não permite a avaliação adequada da presença de IVS ou IVL.

Lymphatic vessel density and lymphatic metastasis in gastroduodenal neuroendocrine carcinoma / 中华内分泌外科杂志

Objective To investigate the relationship of lymphatic vessel density (LVD)and lymphatic metastasis in gastroduodenal neuroendocrine carcinoma.Methods The intratumor lymphatic vessel deusity(iLVD) and peritumor lymphatic vessel density(pLVD) of 55 surgical gastroduodenal neuroendocrine carcinoma tissue samples were detected by immunohistochemistry(with monoclona1 antibody D2-40).The relationship of lymphatic metastasis and pLVD,iLVD was analyzed.Results The value of pLVD and iLVD was 31.02 ± 1.22 vs 5.29 ± 0.76 for gastroduodenal neuroendocrine carcinoma with lymph node metastasis,while it was 20.43 ± 1.39 vs 5.64 ± 1.01 for gastroduodenal neuroendocrine carcinoma without lymph node metastasis,showing pLVD was significantly higher than iLVD in both groups.pLVD of gastroduodenal neuroendocrine carcinoma was closely related to lymph node metastasis (P =0.001,r =0.872),the differentiation of carcinoma,and lymphatic involvement while iLVD of gastroduodenal neuroendocrine carcinoma had no relation with lymph node metastasis(P =0.293).Conclusion Compared to iLVD detection,pLVD detection is more valuable in gastroduodenal neuroendocrine carcinoma since it can help to predict lymph node metastasis status and the prognosis.

Lymphatic vessel density of esophageal adenocarcinoma and its clinical significance / 中华消化外科杂志

Objective To investigate the differences on lymphatic vessel density (LVD) among esophageal adenocarcinoma (EAC),esophageal squamous cell carcinoma (ESCC) and normal esophageal tissues,and analyze the clinical significance.Methods Twenty samples of EAC,24 samples of ESCC and 20 cases of normal esophageal tissues were obtained at the Affiliated Hospital of North Sichuan Medical College from January 2004 to January 2011.D2-40 was used for immunostaining of lymphatic vessels in EAC,and antibodies of D2-40 and Ki-67 were used together to detect proliferation of lymphatic vessels.The differences in the LVD among EAC,ESCC and normal esophageal tissues were analyzed.All data were analyzed using the analysis of variance or t test.Results D2-40 staining could identify the lymphatic vessels,and antibodies of D2-40 and Ki-67 could detect the proliferation of lymphatic vessels.The LVD of EAC,ESCC and normal esophageal tissues were (3.3 ± 1.7)/0.17 mm2,(4.6 ± 1.2)/0.17 mm2 and (3.8 ± 1.2)/0.17 mm2,respectively,with significant differences (F =5.44,P ＜0.05).The LVD of EAC was significantly lower than that of ESCC (t =3.074,P ＜ 0.05),while there was no significant difference in the LVD between the EAC and normal esophageal tissues (t =-1.022,P ＞ 0.05).There were significant differences in the LVD between the ESCC and normal esophageal tissues (t =2.395,P ＜ 0.05).There were significant differences in the LVD between EAC patients with deglutition discomfort and those with pain (t =3.092,P ＜ 0.05).There were significant differences in the LVD between EAC patients with course ＜6 months and those with course≥6 months (t =3.092,P ＜ 0.05).No statistical difference in clinicopathological parameters including gender,age,site of lesion,tumor diameter,pathological morphology,T stage,N stage,G stage,TNM clinical stage and lymph node metastasis were detected (t ＝ 1.130,1.020,F =0.082,t =0.799,F =0.692,t =0.694,1.820,0.353,0.969,0.969,P ＞ 0.05).Conclusions The LVD of EAC is lower than that of ESCC,but is similar to that of normal esophageal tissues.The LVD of EAC is correlated with the symptoms and course of patients.