Cytology and the cell block method in diagnostic characterization of canine lymphadenopathy and in the immunophenotyping of nodal lymphoma

Minimally invasive techniques used to evaluate canine peripheral lymphadenopathy (PLN), including fine needle aspiration biopsy with cytological evaluation (FNAB‐C) and flow cytometry (FC), have benefits and limitations. The cell block (CB) method is an alternate processing technique in which fine needle aspirate biopsy samples are concentrated, fixed, and embedded in paraffin for routine histological processing/staining. Utilizing three observers, we determined the diagnostic value of the CB in evaluating canine PLN across six categories (non‐diagnostic, reactive, inflammatory/infectious, probable lymphoma and lymphoma, metastatic neoplasia) and correlated findings to immunophenotypic and clonal antigen receptor rearrangement results in canine nodal lymphoma. Eighty‐five paired FNAB‐C and CB samples were evaluated from canine patients presenting to the University of Minnesota Veterinary Oncology or Internal Medicine services. Diagnostic quality samples were obtained in 55/85 (65%) CB and 81/85 (95%) FNAB‐C samples, respectively, and nodal pathology impacted CB diagnostic yield. Overall percent agreement between diagnostic‐quality FNAB‐C and CB samples was 86%, but increased to 95% if the categories of lymphoma and probable lymphoma were combined. There was 100% agreement for both the diagnoses of metastatic neoplasia and reactive lymph nodes and 92% agreement for the diagnosis of lymphoma/probable lymphoma. Using immunohistochemistry (IHC), CB samples correctly immunophenotyped 22/23 (96%) cases of B‐cell lymphoma, but only 1/6 (17%) cases of T‐cell lymphoma. IHC was not completed on nine cases of lymphoproliferative disease because of insufficient cellularity. When the CB method (CBM) yielded diagnostic quality samples there was good to excellent agreement with FNAB‐C samples and CB samples were suitable for some IHC tests.     

Relation between age, body weight, and medial retropharyngeal lymph node size in apparently healthy dogs.

The aims of this study were to determine the size of the medial retropharyngeal lymph nodes in apparently healthy dogs using ultrasonography and to investigate relationships between body weight (1.8-59 kg), age (1.0-15 years), and medial retropharyngeal lymph node sizes (width, height, and length). The sample population consisted of 100 apparently healthy, volunteered, adult dogs. The data were normally distributed, thus mean, SD, and Pearson's correlation were used. Repeatability of ultrasound measures was assessed as the percentage of differences between duplicate measures that were within 2 SDs of the differences: all measures were at least 93% repeatable (differences typically were < or = 0.25 cm and always < 1 cm). No difference between sexes was observed. The medial retropharyngeal lymph node increased in size with increased body weight (r = 0.46 to 0.59) and decreased in size with increased age (r = -0.30 to -0.50). Although statistically significant, the actual variation is not likely clinically important due to the small range of sizes, measurement error, and various combinations of age and body weight. Therefore, regardless of body weight or age, the average width is 1.0 cm, height is 0.5 cm, and length is 2.5 cm and maximum width is 2 cm, height is 1 cm, and length is 5 cm. Based on the maximal difference between duplicate measures (with some exception), any change > or = 0.4 cm with width or height, or > or = 1.0 cm in length, in a follow up measurement probably represents a true biological change rather than measurement error.     

Morphometrical approach for predicting regional lymph node micrometastatic load in canine mast cell tumours: preliminary results

Canine cutaneous mast cell tumours (MCTs) have a variable biologic behaviour, and accurate staging is necessary to dictate therapy and predict outcome. Regional lymph node (RLN) involvement is a relevant prognostic factor. While obvious lymph node (LN) metastases are relatively easy to be diagnosed, micrometastatic disease recognition is challenging. The main aim of the study was to evaluate the number of mast cells (MCs) in the LNs of clinically healthy dogs ( n = 4, group 1), dogs with inflammatory diseases ( n = 31, group 2) and dogs with cutaneous MCT ( n = 27, group 3), including animals with no RLN metastases (subgroup 3.1), those with occasional MCs in RLNs (3.2) and those with obvious RLN metastasis (3.3). MCs also were morphometrically evaluated for the following nuclear parameters: mean nuclear area (MNA), mean nuclear perimeter (MNP), largest to smallest diameter length (LS ratio), mean nuclear form factor and coefficient of variation of nuclear area. The average percentages of MCs were 0.0 and 0.01 in groups 1 and 2, respectively, and 0.07, 2.4 and 47.1 in subgroup 3.1, 3.2 and 3.3. MNA and MNP were significantly higher in subgroup 3.3 than in group 2 ( P < 0.05). MNA and MNP in subgroup 3.2 suggested the presence of neoplastic MCs; this prediction of micrometastatic load correlated with outcome. Analysis of preliminary results shows that nuclear morphometry is useful to detect micrometastatic disease in RLN of dogs bearing cutaneous MCTs.     

The role of sentinel lymph node mapping in small animal veterinary medicine: A comparison with current approaches in human medicine

The relevance of regional lymph node (LN) assessment to quantify the metastatic spread of cancer is well recognized in veterinary oncology. Evaluation of LNs is critical for tumour staging. However, sampling the correct LN may not be possible without sentinel lymph node (SLN) mapping. Methods for diagnostic imaging and intraoperative detection of SLNs are well established in human medicine, in particular, the combination of lymphoscintigraphy and intraoperative application of blue dyes. Nevertheless, alternative imaging techniques are available and have gained increasing interest. Successful implementation of these techniques in dogs have been reported in both clinical and experimental studies. This review aims to provide an overview of SLN mapping techniques in human and veterinary medicine.     

The sonographic evaluation of lymph nodes

Ultrasound can be used to detect and evaluate both normal and abnormal lymph nodes, as well as aid in biopsy sampling procedures, an important part of staging procedures in cancer patients. Several parameters can be evaluated using ultrasound; lymph node size, margins, echogenicity, echopattern (echotexture), acoustic transmission, presence and distribution of vascular flow, and vascular flow indices. The most diagnostically helpful include the short/long axis ratio of the lymph node, the pattern of distribution of the blood vessels within the lymph node, and to some extent the resistive and pulsatility indices. This review discusses the use of ultrasound for detecting, evaluating, and sampling peripheral, abdominal and thoracic lymph nodes in small animals.     

Direct Detection of Mycobacterium bovis in Bovine Lymph Nodes by PCR

Thirty‐five lymph node samples were taken from animals with macroscopic lesions consistent with Mycobacterium bovis infection. The animals were identified by postmortem examination in an abattoir in the northwestern region of state of Paraná, Brazil. Twenty‐two of the animals had previously been found to be tuberculin skin test positive. Tissue samples were decontaminated by Petroff’s method and processed for acid‐fast bacilli staining, culture in Stonebrink and Lowenstein‐Jensen media and DNA extraction. Lymph node DNA samples were amplified by PCR in the absence and presence (inhibitor controls) of DNA extracted from M. bovis culture. Mycobacterium bovis was identified in 14 (42.4%) lymph node samples by both PCR and by culture. The frequency of PCR‐positive results (54.5%) was similar to that of culture‐positive results (51.5%, P > 0.05). The percentage of PCR‐positive lymph nodes increased from 39.4% (13/33) to 54.5% (18/33) when samples that were initially PCR‐negative were reanalysed using 2.5 μl DNA (two samples) and 1 : 2 diluted DNA (three samples). PCR sensitivity was affected by inhibitors and by the amount of DNA in the clinical samples. Our results indicate that direct detection of M. bovis in lymph nodes by PCR may be a fast and useful tool for bovine tuberculosis epidemic management in the region.     

磁共振常规及弥散加权成像对预测宫颈癌淋巴结转移的价值

目的 探讨磁共振常规及弥散加权成像在宫颈癌淋巴结转移诊断中的敏感性、特异性.方法 113例ⅠB1至ⅡB期宫颈癌患者行术前MRI常规及弥散加权成像检查、根治性子宫切除术加盆腔或盆腔及腹主动脉旁淋巴结清扫术.结果 94例(83.2%)经术后病理学检查证实无淋巴结转移,19例(16.8%)有淋巴结转移.MRI常规及弥散加权成...     

SONOGRAPHIC CHARACTERISTICS OF PRESUMPTIVELY NORMAL CANINE MEDIAL ILIAC AND SUPERFICIAL INGUINAL LYMPH NODES

The medial iliac and superficial inguinal lymph nodes are not routinely palpable in the dog, and ultrasound imaging provides an alternate noninvasive technique to assess these lymph nodes, as well as to guide needle aspiration. Herein we describe the ultrasound characteristics of the medial iliac and superficial inguinal lymph nodes in 50 healthy dogs, as well as frequency and ease of node detection. The relationship between the size of the lymph nodes and the following variables was assessed: age, gender, body weight, body condition score, body length, and thoracic height and width. Right and left medial iliac lymph nodes were detected in 50 (100%) dogs, right superficial inguinal lymph node(s) in 49 (98%) dogs, and left superficial inguinal lymph node(s) in 47 (96%) dogs. In >90% of both sets of lymph nodes, the echogenicity was hypoechoic or isoechoic to surrounding tissues, with a corticomedullary or homogenous echotexture, smooth, clearly defined margins, and a fusiform shape. Increasing weight, distance from the sternal manubrium to the ischium, and thoracic height and width were associated with increased lymph node size (P‐values<0.05). Average lymph node sizes and range of sizes provide preliminary reference values for the medial iliac and superficial inguinal lymph nodes in normal dogs.     

Mesenteric lymph drainage alleviates spleen injury in hemorrhagic shock rats

AIM：To observe the effects of post-shock mesenteric lymph (PSML) drainage on histopathology, apoptosis, cell cycle and proliferation of the spleen in rats with hemorrhagic shock. METHODS：Eighteen Wistar rats were randomly divided into sham, shock and shock+drainage groups (n=6 in each group). The hemorrhagic shock model was established in the shock and shock+drainage groups. Fluid resuscitation for 30 min was performed 1.5 h after hypotension, and PSML was drained in the rats in shock+drainage group from 1 h after hypotension to 3 h after resuscitation finished. The fixed spleen tissue was harvested from each rat for histological observation with HE staining. The apoptosis of splenocytes was observed by Hoechst 33258 staining. The expression of Bcl-2 and Bax proteins was detected by immunohistochemical staining. The cell cycle and the expression of p53 protein were measured by flow cytometry, and the proliferation index (PI) was calculated. RESULTS：Compared with sham group, splenic tissue injury appeared in the shocked rats. The apoptotic cells and the expression of Bax and p53 in shock group were increased, while Bcl-2 expression was decreased. The percentage of G2/M cells in shock group was decreased. Compared with shock group, the splenic tissue damage in shock+drainage group was significantly attenuated. Moreover, the number of apoptotic cells, the percentage of G0/G1 cells, and the expression of Bax and p53 were obviously decreased, and the G2/M cells, Bcl-2 protein expression and PI were significantly increased in shock+drainage group. CONCLUSION: PSML drainage alleviates splenic injury in hemorrhagic shock rats, which may be related to reducing the apoptosis of splenocytes.     

Role of hydrogen sulfide in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats

AIM：To investigate the role of hydrogen sulfide (H2S) in alleviation of liver injury by mesenteric lymph drainage in hemorrhagic shock rats. METHODS：A hemorrhagic shock model was established in male Wistar rats. DL-propargylglycine (PPG), an inhibitor of cystathionine γ-lyase (CSE) which is a synthase of H2S, or sodium hydrosulfide (NaHS), a donor of H2S, was administered to the hemorrhagic shock rats with mesenteric lymph drainage. The rats were randomly divided into sham, shock, shock+drainage, shock+drainage+PPG (45 mg/kg, ip, 0.5 h before hemorrhage) and shock+drainage+NaHS (28 μmol/kg, ip, 0.5 h before hemorrhage) groups. Fluid resuscitation was performed 1 h after hypotension, and then mesenteric lymph was drained in the rats of shock+drainage, shock+drainage+PPG and shock+drainage+NaHS groups for 3 h. The hepatic histomorphology was observed. The biochemical indexes of hepatic function in plasma, and H2S, CSE, Toll-like receptor 4 (TLR4), interleukin (IL)-10, IL-12 and tumor necrosis factor α (TNF-α) in hepatic homogenate were also examined. RESULTS：The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bile acid (TBA) in plasma, and H2S, CSE, TLR4, IL-10, IL-12 and TNF-α in hepatic homogenate in shock group were significantly higher than those in sham group. Mesenteric lymph drainage obviously reduced these indexes in shock rats, except for TLR4. PPG further decreased these indexes except for CSE, while NaHS increased these indexes except for TBA and CSE. Morphological observation showed that liver injury appeared in the rats from shock and shock+drainage+NaHS groups, and there was nearly normal hepatic structure in the rats from sham, shock+drainage and shock+drainage+PPG groups. CONCLUSION：The mechanism of mesenteric lymph drainage alleviating liver injury in hemorrhagic shock rats is related to reducing the production of H2S and alleviating the H2S-mediated inflammation.