Purpose:This study aimed to monitor the presence of enzootic nasal tumor virus 2 (ENTV-2) and the dynamic changes of the viral env gene during the successive passages of enzootic nasal adenocarcinoma (ENA) cells. Materials and Methods:ENA cells were passaged for 24 generations, and the supernatants and cell pellets were collected to extract RNA for monitoring viral presence. The ENTV-2 env gene was amplified by PCR, cloned, and sequenced to analyze nucleotide mutations in the env gene sequence across different passages. Results:The results showed that ENTV-2 RNA was consistently detected in ENA cells up to the 22nd passage, indicating sustained viral persistence within the cells, while its presence in the supernatant was unstable, suggesting possible regulation of viral release, although the underlying mechanism was not investigated. Further cloning and sequencing of the env gene from 11 passages revealed no deletions or insertions, but point mutations were observed, predominantly A ⟶ G substitutions, some of which led to amino acid changes. Notably, nonsense mutations W231 and W19 occurred in the 7th and 15th passages, respectively, resulting in premature truncation of the Env protein, while the Q168R substitution was consistently identified in the sequenced clones from all passages from Passage 5 through Passage 22. Structural predictions indicated that the Env protein maintained a conserved core structure with localized variations in non-transmembrane regions. These observations are consistent with possible structural fine-tuning, but functional validation is required. Conclusion:This study provides a descriptive account of sequence changes in the ENTV-2 env gene during serial passaging of ENA cells. The findings are hypothesis-generating and await experimental verification of their biological significance.

A 5-month-old male Maine Coon cat was presented to the hospital with persistent salivation for 2 weeks. Serum biochemistry showed markedly elevated levels of blood ammonia and bile acids. An abdominal computed tomography discovered a shunt from the left gastric to the left phrenic vein, and the right ureter was seen to descend to the level of L4 and L5 normally and then pass dorsally and medially to the caudal vena cava, thus forming a loop. The present case, in which a circumcaval ureter and a congenital extrahepatic portosystemic shunt were diagnosed concurrently in a cat, was extremely rare and was subsequently confirmed by surgery. Based on the reviewed literature, this is the first case report of the concurrence of a circumcaval ureter and a congenital extrahepatic portosystemic shunt in a cat.

Introduction:Enzootic nasal adenocarcinoma (ENA) is a nasal cancer that occurs in goats and sheep infected by enzootic nasal tumour virus. Pathologic examinations are useful for distinguishing tumours from inflammatory hyperplasia. The aim of this study was to describe the pathological characteristics of ENA. Material and Methods:Caprine tumour samples were collected for pathological examination. The tissue sections were stained with haematoxylin-eosin and periodic acid-Schiff (PAS) and processed for immunohistochemical staining. Tumour samples were also processed for routine transmission electron microscopy (TEM). Results:The histopathological structure of the tumours exhibited both papillary formations in the superficial regions and tubular or acinar formations in the deeper layers, representing distinct structural patterns within the same adenocarcinoma. The tumour cells were positive for PAS, and mitotic figures were rare. Low-differentiated cancer nests and epithelial-mesenchymal transition phenomena were observed. Immunohistochemical analysis showed that the tumour cells were strongly positive for pancytokeratin and cytokeratin (CK)18, moderately positive for CK7, and did not express olfactory marker protein. The Kiel 67 labelling index was approximately 23%. Retrovirus-like particles were distributed inside and outside of acinar tumour cells in TEM. Conclusion:The origin site of ENA is the epithelium of the nasal glandular tubules. This cancer is a low-grade adenocarcinoma with malignant potential. Cytokeratin 7 and CK18 can be considered immunophenotypes for identifying ENA tumour cells.

Lymphography is a useful technique in the diagnosis of lymphatic diseases. Conventional lymph node imaging methods, such as subcutaneous and footpad injections, are invasive and unable to visualize mesenteric lymph nodes. Liposomes have the potential to increase the oral bioavailability of poorly bioavailable hydrophilic drugs and promote their lymphatic transport in the intestinal lymph. In this study, the iohexol liposome was prepared using cholesterol, soybean lecithin, and iohexol by the reverse-phase evaporation method. It had an encapsulation efficiency of 70.26%, an average particle size diameter of 185.7 nm, a polydispersity index of 0.275, and a zeta potential of -7.697 mV. The iohexol liposomes were stable for five days at 4 °C under static conditions. The iohexol content in the lymph nodes of mice after oral iohexol liposomes initially increased and then gradually decreased over time, with the absorption peak occurring around 50-70 minutes and a peak iohexol content of 5.09 mg/g. After oral administration of iohexol liposomes, mild enhancement (12.46-12.92 HU) was observed in the mesenteric lymph nodes of dogs through CT scanning after a certain period. These results indicate that iohexol liposomes, when administered orally, can effectively achieve imaging of the mesenteric lymph nodes. Overall, we provide a novel noninvasive imaging modality based on liposomes for evaluating mesenteric lymph nodes via lymphography.

Enzootic nasal adenocarcinoma (ENA) is a contagious tumor disease of goats and sheep, which is caused by enzootic nasal tumor virus (ENTV). To better understand the pathogenesis of ENA, this study aimed to establish a goat ENA cell line (ENA-1). The cells have been characterized with regard to morphology, growth rate, ultrastructural features, chromosome number, expression of CK7 and CK18, tumorigenicity, species, and mycoplasma contamination. ENA-1 had an epithelioid cell morphology with an unstable chromosome number under a light microscope. Under an electron microscope, the cell nuclear heterogeneity was not obvious, and there were more intermediate filaments and a small number of immature retrovirus-like particles in the cytoplasm. ENA-1 had strong proliferative potential, and the cell multiplication time was about 36 h, which could make BALB/c nude mice develop tumors. CK7 and CK18 were expressed in the cytoplasm of primary goat tumors, in transplanted tumors from nude mice, and un ENA-1 cells with the same intensity. PCR revealed that ENA-1 continuously carried ENTV-2 up to the 17th generation with no germline contamination or mycoplasma contamination. In conclusion, using a serum-containing culture system, ENA-1 cells were successfully isolated, cultured, and purified from goat tumor tissues. The isolated ENA-1 cells retained robust proliferation potential and maintained their phenotype, indicating the potential application of the ENA-1 cell line as an in vitro model of ENA.

This study aimed to explore the magnetic resonance cholangiopancreatography (MRCP) manifestations of the pancreaticobiliary ducts and assess pancreatic exocrine function using Secretin-enhanced MRCP (S-MRCP) in dogs with acute edematous pancreatitis (AEP). Twelve (12) dogs were included, with six in the AEP group and six in the control group. The AEP model group underwent a procedure by way of pancreatic duct retrograde injection. The control group underwent the same procedure without infusion. MRCP and S-MRCP were performed identically in both the AEP group and control groups. MRCP was performed pre-surgery and on the 1st, 3rd, and 5th days post-surgery in the AEP group and control groups. The pancreaticobiliary duct diameter was significantly wider in the AEP group compared to the control group (P<0.01) on the 5th day. The S-MRCP images were evaluated on the 3rd day post-surgery at 1, 3, 5, 7, and 11 min after secretin infusion. Compared to the control group, the pancreatic ducts in the AEP group became dilated and branched which appeared, reaching the largest infusion, the degree of duodenal filling in the AEP group was Grade 2, indicating impaired pancreatic exocrine function in dogs with AEP. In conclusion, MRCP provided excellent comprehensive visualization of the pancreatic and hepatobiliary duct systems in dogs with AEP. S-MRCP effectively detected branched pancreatic ducts and impaired pancreatic exocrine in AEP dogs.

探究犬胰腺磁共振成像(magnetic resonance imaging,MRI)扫描序列总结归纳犬急性水肿型胰腺炎(acute edematous pancreatitis,AEP)的MRI影像表现.选用10只成年本地杂种犬为研究对象,手术经副胰管逆行注入牛磺胆酸钠与胰蛋白酶混合液建立犬急性水肿型胰腺炎模型,然后进行MRI扫描.MRI多序列扫描图像主观与客观分析结果显示,门控采集快速自旋回波T2加权成像(FSE-T2WI-TRIG)序列与门控采集脂肪抑制快速自旋回波T2加权成像(FSE-T2WI-FS-TRIG)序列评分较高,可作为犬胰腺首选MRI扫描序列,必要时可进行造影增强扫描.FSE-T2WI-TRIG、FSE-T2WI-FS-TRIG序列AEP表现为胰腺体积呈弥散性增大,外形不规则,胰腺实质T2WI信号呈不均匀增高,强度高于同层肝组织信号.胰腺与周围脂肪界限稍有不清,胰腺周围伴有渗出,呈条片状T2WI高信号.造影后胰腺实质不均匀强化,平均强化峰值时间为注射造影剂后60～90 s,存在明显延迟强化.研究结果表明,本研究筛选的MRI扫描序列可用于临床上犬AEP的检查,同时AEP的MRI影像表现可为临床上犬AEP的诊断提供重要的依据.

旨在筛选犬胰腺常规磁共振成像(MRI)扫描序列,获得犬胰腺的MRI影像解剖结构图谱.利用1.5特斯拉(1.5T)磁共振扫描10条健康本地杂种犬,采用梯度回波(GRE)序列、快速自旋回波(FSE)序列、脂肪抑制(FS)、磁共振胰胆管造影(MRCP)、平衡稳态自由进动梯度回波序列(GRE_BSSFP)、弥散加权成像序列(EPI_DWI)对犬胰腺的横断面和冠状面进行了扫描;对所获取的图像进行详尽的解剖学注释,对比了不同成像序列中犬胰腺、胰管等组织结构的显影效果,分析不同序列获得的MRI图像.推荐临床犬胰腺常规MRI扫描序列:横断面FSE、FSE_FS、GRE、EPI_DWI,冠状面FSE_FS,横断面对比增强T1WI,依此得到了一套较为完整的犬胰腺MRI影像解剖图谱.本研究结果可为犬胰腺疾病的影像学诊断提供参考.

确定猫肾脏CT多期增强扫描最佳造影剂使用剂量、注射速率及延迟时间.选择不同剂量的碘海醇(350,400,450,500 mg I/kg)及注射速率(3,4 mL/s)对猫肾脏进行造影,动态扫描测量主动脉、肾皮质CT值,计算主动脉、肾皮质CT增强值(ΔCT),通过比较主动脉及肾皮质ΔCT的变化确定最佳造影条件.使用最佳造影条件对猫肾脏进行造影,动态扫描并绘制肾脏时间—密度曲线,统计肾皮质达峰时间TCP、肾皮质与肾髓质CT值相近的起始时间TNP、观察造影剂排泄至肾盂和膀胱的情况,并计算肾皮质达峰时间与注射时间的差值ΔTCP、肾皮质与肾髓质CT值相近的起始时间TNP与T注射的差值ΔTNP,最终确定各期最佳扫描延迟时间.试验结果表明:最佳造影剂剂量为450 mg I/kg、注射速率3 mL/s,该条件下主动脉及肾皮质ΔCT值较高.计算得出ΔTCP平均为11 s,ΔTNP平均为26 s,各期扫描延迟时间分别为:T皮髓期延迟=T注射+10s,T实质期延迟＝T注射+26 s,T排泄期延迟=2 min,T膀胱期延迟=4 min.按照上述条件可获得满意的猫肾脏CT多期增强扫描图像.

(目的)为探讨犬椎间盘疾病的磁共振诊断方法和手术治疗价值.(方法)对8例犬进行体格检查、神经学检查和磁共振检查,确诊患有椎间盘疾病,并手术治疗.(结果)磁共振图像显示患犬椎间盘脱水、钙化,呈T2WI低信号,并向背侧膨出或突出,压迫脊髓.脊髓受压水肿,呈T2WI高信号.确诊患有椎间盘疾病.行半椎板切除术或背侧椎板切除术进行手术治疗,6只犬预后良好,2只犬预后谨慎.(结论)磁共振检查可以清晰的显示椎间盘疾病,手术治疗根据发病情况预后不定.

采用转录组学测序的方法筛选三氮脒药物处理后犬吉氏巴贝斯虫差异表达基因.采集感染犬吉氏巴贝斯虫病犬的血液,用终浓度为800 nmol/L的三氮脒处理24 h,提取RNA进行转录组测序,比较三氮脒处理后犬吉氏巴贝斯虫基因表达的差异.对测序结果进行无参转录组分析,经筛选共获得77个巴贝斯虫属的基因,将筛选后的基因进行GO和KEGG数据库分析,其注释富集基因最多的类别是细胞组分、分子功能和生物进程,在KEGG通路注释中,显示与翻译通路有关的基因较多.对三氮脒处理组与未处理组的样品进行组间差异基因表达分析,发现基因CL118转录水平显著性下调.采用real-time PCR方法进行验证,结果显示本实验室保存虫株以及临床野生虫株经三氮脒处理24 h后,CL118基因的转录水平均发生下调.结果表明,基因CL118可能是三氮脒药物作用于犬吉氏巴贝斯虫虫体的潜在作用靶点.

旨在检测患病犬经三氮脒治疗后,吉氏巴贝斯虫线粒体细胞色素b(Cytb)是否存在基因突变.调查了 2010至2020年犬巴贝斯虫病的流行情况,收集了 11份自然感染吉氏巴贝斯虫且接受三氮脒治疗的患犬血液样品,采用蛋白酶K消化,苯酚提取法提取血液中基因组DNA,设计引物,通过PCR扩增吉氏巴贝斯虫Cyt b基因片段并进行测序分析.调查发现,自2010年以来,通过三氮脒治疗的吉氏巴贝斯虫病的疗程逐渐延长,复发率也逐渐增加.在11份血样中,吉氏巴贝斯虫的Cytb基因彼此之间相似性高达99.65％～100％,与日本分离株也有较高的同源性(99.18％～99.54％).在Cytb基因nt363基因位点上没有发现基因突变.这些结果说明虽然吉氏巴贝斯虫对三氮脒逐渐产生耐药,但三氮脒对吉氏巴贝斯虫Cyt b基因并不产生影响.

New antibabesial drugs are required to fight resistant parasites, and plant-derived natural products are a robust source. Six kinds of natural product extracts derived from herbal medicines that are traditionally used for the treatment of malaria were selected to test the antibabesial effect on Babesia gibsoni in vitro and in vivo. Parasitized blood was collected from dogs infected with B. gibsoni to evaluate the inhibitory effect of verbenalin, catechin hydrate, dihydrolycorine, embelin, ursolic acid, agrimol B, and bruceine H in vitro. The expression levels of the 18S rRNA gene in all drug-treated groups were determined by relative quantification using a real-time PCR method. Significant inhibition of the in vitro growth of B. gibsoni was observed after treatment by those natural product extracts (200 nM concentration) (P < 0.05). Catechin hydrate showed the highest activity in vitro due to the lowest expression levels of the 18S rRNA gene. The IC50 value of catechin hydrate against B. gibsoni was 273 nM. In B. gibsoni infected dogs, intravenous administrations of catechin hydrate and diminazene aceturate showed significant (P < 0.05) inhibition of B. gibsoni growth at a dose of 11 mg/kg and 10 mg/kg, respectively, compared to the control group. The results of our study may suggest that catechin hydrate may be a promising alternative to treat canine babesiosis caused by B. gibsoni.

本试验旨在确定犬肝多期增强扫描造影剂使用剂量、注射速率及最佳延迟时间.选取不同的碘海醇剂量(500、575、650 mg·kg-1,以I含量计)及速率(2、3 mL·s-1)对犬进行造影,动态扫描,计算造影前后主动脉、门静脉、肝实质CT增强值,确定最佳造影剂剂量及注射速率.然后采用最佳造影剂剂量和注射速率对不同体型的犬进行造影,动态扫描后绘制时间-密度曲线,统计主动脉、门静脉、肝实质的达峰时间,计算达峰时间和注射时间的差值(△tAO/△tsp/△tL),确定各期最佳扫描延迟时间.研究结果显示,当采用575 mg·kg-1、3 mL·s-1的造影剂剂量和注射速率时得到的主动脉、门静脉、肝实质CT增强值较高,可获得较好的增强效果.通过时间-密度曲线分别计算小、中、大3种体型犬的△tAO分别为7、9、4 s,△tsp分别为21、23、17 s,△tL分别为41、44、34 s,各期最佳扫描延迟时间可用公式"注射时间+△tRoi—1/2扫描时间"计算得到.通过临床病例验证,本试验使用的造影剂剂量(575 mg·kg-1)、注射速率(3 mL·s-1)及延迟时间("注射时间+△tAo/△tsp/△tL-1/2扫描时间")临床效果较好,可应用于犬肝疾病的CT造影检查.

The aim of this study was to describe radiography, computed tomography (CT), and magnetic resonance imaging (MRI) findings of enzootic nasal tumors in goats infected with enzootic nasal tumor viruses. Five of six goats with a mean age of 2 years, showed clinical signs of respiratory disease. Head radiographs showed increased density of the unilateral or bilateral nasal cavity in four goats, and a CT scan showed that the space-occupying lesion of the nasal cavity originated from the ethmoid bone and was enhanced homogeneously postcontrast in all goats. The nasal concha was destroyed and the paranasal sinus mucosa was thickened and filled with fluid in some goats. On MRI, the mass exhibited equal or slightly higher signal intensity on T2 weighted images, equal signal intensity on T1 weighted images, a high signal on fluid-attenuated inversion recovery images and heterogeneous enhancement postcontrast. After dissection, histopathological examination of the mass and virus genome detection of the nasal secretions confirmed that the intranasal mass was a low-grade adenocarcinoma and that the goats were infected with enzootic nasal tumor virus type 2. In conclusion, CT and MRI have high diagnostic values for enzootic nasal tumors because they match the postmortem findings and are more accurate than radiography.

Dried blood spot (DBS) based PCR was considered an inexpensive and feasible method for detecting pathogens in the blood. The DBS carrier filter paper and PCR kits are crucial for accurate diagnosis. We evaluated 4 types of filter papers and 20 PCR kits for DBS samples. The PCR detecting Plasmodium results showed that the minimum detection limit of the 4 filter papers was 1 × 102 parasites/μL, and the positive rates of 20 PCR kits ranged from 0% to 100%. PCR results were satisfactory for detecting Plasmodium falciparum (P. falciparum) and Plasmodium. vivax (P. vivax) in archived DBS samples and Babesia gibsoni (B. gibsoni) in fresh pet DBS samples. Our results provided a useful reference for the detection of blood pathogens with DBS samples and direct PCR, especially for screening the cost-efficacy combination of filter paper and PCR kit in resource-limited areas.

为了明确吉氏巴贝斯虫感染后犬脾脏的病理学变化,本试验采用静脉接种的方法感染健康犬,每只犬接种含吉氏巴贝斯虫的血液5 mL(红细胞染虫率约2％),接种后每3天采集1次血液,通过血涂片、血常规、PCR等方法监测吉氏巴贝斯虫是否感染成功;每7天采用B超检查脾脏的大小;对发病死亡犬进行病理剖检和组织病理学检查,观察脾脏病理组织结构的变化.结果显示,人工接种吉氏巴贝斯虫3 d后PCR可检测到虫体DNA,6d后血涂片可见虫体,15 d后血常规指标(红细胞总数、血红蛋白含量和红细胞压积)开始降低.B超检查结果显示,犬感染吉氏巴贝斯虫后其脾门、脾尾厚度随病程延长逐渐增大.于感染后第31、36天和第42天试验犬分别死亡1只,死亡率为50％,剖检死亡犬可见其皮下黏膜黄染,脾脏肿大,被膜紧张,边缘钝圆,脾梗死.组织病理学观察可见脾脏充血,白髓、红髓界限不清,大量中性粒细胞、单核巨噬细胞浸润,结缔组织增生,脾小梁增粗,梗死灶内大面积坏死,淋巴细胞减少,中性粒细胞浸润.由此可见,在吉氏巴贝斯虫感染过程中脾脏首先发生急性炎性脾肿,随着病程延长逐渐演变成坏死性脾炎和慢性脾炎.

旨在分析犬椎间盘疝出疾病的发病情况及相关的风险因素,为临床上犬椎间盘疝出疾病的诊断、治疗和预防提供参考.本研究收集了 147例经临床体格检查、神经学检查、核磁共振影像检查综合确诊为椎间盘疝出的病例,回顾性分析了犬椎间盘疝出的类型、发病部位,以及与动物品种、性别、年龄之间的关系.结果显示,4种椎间盘疝出类型中,椎间盘脱出发病数最多,占43.9％;椎间盘疝出部位中,C6-T2脊髓节段发病数最少,T3-L3脊髓节段发病数最多;综合分析动物品种、发病年龄和椎间盘疝出类型,软骨发育不良品种的椎间盘疝出更易发生于成年期,且以椎间盘脱出为主,而其他犬种椎间盘疝出更易发生于老年,以椎间盘突出为主.本研究综合分析椎间盘疝出疾病的发病特点和流行病学特征,填补国内研究空白,为该病临床诊疗提供参考资料.

Tumorigenesis is associated with metabolic abnormalities and genomic instability. Microsatellite mutations, including microsatellite instability (MSI) and loss of heterozygosity (LOH), are associated with the functional impairment of some tumor-related genes. To investigate the role of MSI and LOH in sporadic breast tumors in canines, 22 tumors DNA samples and their adjacent normal tissues were evaluated using polyacrylamide gel electrophoresis and silver staining for 58 microsatellites. Quantitative real-time polymerase chain reaction, promoter methylation analysis and immunohistochemical staining were used to quantify gene expression. The results revealed that a total of 14 tumors (6 benign tumors and 8 breast cancers) exhibited instability as MSI-Low tumors. Most of the microsatellite loci possessed a single occurrence of mutations. The maximum number of MSI mutations on loci was observed in tumors with a lower degree of differentiation. Among the unstable markers, FH2060 (4/22), ABCC9tetra (4/22) and SCN11A (6/22) were high-frequency mutation sites, whereas FH2060 was a high-frequency LOH site (4/22). The ABCC9tetra locus was mutated only in cancerous tissue, although it was excluded by transcription. The corresponding genes and proteins were significantly downregulated in malignant tissues, particularly in tumors with MSI. Furthermore, the promoter methylation results of the adenosine triphosphate binding cassette subfamily C member 9 (ABCC9) showed that there was a high level of methylation in breast tissues, but only one case showed a significant elevation compared with the control. In conclusion, MSI-Low or MSI-Stable is characteristic of most sporadic mammary tumors. Genes associated with tumorigenesis are more likely to develop MSI. ABCC9 protein and transcription abnormalities may be associated with ABCC9tetra instability.