Tumour necrosis factor stimulated gene 6 intrinsically regulates PD-L1 expressions in breast cancer cells,leading to modulation of tumour microenvironment

Recent studies have shown that tumour cells express tumour necrosis factor-inducible gene 6 (TSG-6) and its protein, which is known to play a key role in regulating excessive immune responses and proliferation and growth of mesenchymal stem cells (MSCs). It has not been confirmed whether the inhibition of TSG-6 for tumour cells can suppress tumour cell growth and regulate the activation of immune cells in the tumour microenvironment (TME). TSG-6-specific small interfering RNA was transfected into canine and human breast cancer cells (CIPp, CIPm and BT-20). TSG-6-down-regulated (siTSG-6) cells showed decreased cell proliferation, migration, and invasion abilities. Decreased mRNA expressions of NF-κB, STAT3 and Sox2, confirming that TSG-6 is an upper factor governing tumour growth and metastasis. Notably, siTSG-6 cells showed significantly decreased expression levels of CD44 and PD-L1. Direct and indirect co-culture of canine peripheral blood mononuclear cells (cPBMCs) and the siTSG-6 cells showed significant activation in M1 type macrophages and cytotoxic T cells. They also showed a tendency to decrease in the expression of CTLA-4 and increase in the expression of PD-1. In conclusion, this study suggests that the down-regulation of TSG-6 in breast cancer cells could not only suppress tumour growth and metastasis, and but also regulate TME. Since modulation of immune checkpoint proteins occurs in both tumour cells and immune cells, inhibiting TSG-6 and its protein within the TME could be novel therapeutic target for anticancer treatment.     

双低菜籽粕对草鱼和鲤甲状腺、肝、肾组织结构的影响

以组织学方法,研究了投喂不同含量双低菜籽粕对草鱼和鲤甲状腺、肝、肾组织的影响。结果表明:在试验的前28 d,双低菜籽粕各不同含量的日粮(17%、34%、51%、68%)对草鱼甲状腺、肝脏组织没有影响,但菜籽粕含量为68%的试验组草鱼肾小管上皮出现颗粒变性。第42天后,菜籽粕含量为68%的试验组草鱼甲状腺滤泡上皮轻度增生,滤泡结构不规则;菜籽粕含量等于或大于51%的试验组草鱼肝脏出现颗粒变性和空泡变性,肾小球有透明变性,肾脏轻度淤血。在试验的第28天,双低菜籽粕含量等于或大于52.2%的试验组鲤甲状腺滤泡上皮细胞的高度增加,肝细胞空泡变性,有轻微的肾小球透明变性。第42天后菜籽粕含量等于或大于34.8%的试验组鲤甲状腺滤泡上皮细胞的高度增加,滤泡胶质的边缘出现缺刻;肝脏出现淤血,部分肝细胞坏死;肾小球透明变性,肾脏淤血。     

INTRA- AND INTEROBSERVER VARIABILITY OF ULTRASONOGRAPHIC MEASUREMENTS OF THE THYROID GLAND IN HEALTHY BEAGLES

The repeatability of ultrasonographic measurements of the canine thyroid gland was evaluated. The variability of three different parameters (the maximal length, width, and height) within observer, between observer and between dogs was assessed based on three different measurements made by each of three observers infive healthy beagle dogs. From the three parameters, the volume of the gland was estimated using a formula of a rotation ellipse. The height and the volume had the lowest intra- and interobserver variability, while measurements of the length had the biggest intra- and interobserver variability. The mean values, with their 90% confidence interval were: height = 0.53cm [0.33-0.73], length = 2.45cm [2.04-2.85], width = 0.62cm [0.46- 0.78], volume = 0.38 cm3 [0.20-0.55].     

Intrathymic inoculation of liver specific antigen protects hepatocytes from apoptosis after liver allotransplantation

AIM: To study the effects of intrathymic inoculation of liver specific antigen (LSA) on hepatocyte apoptosis after liver allotransplantation. METHODS: Orthotopic liver transplantation was used in this study. Group Ⅰ: syngenic control (Wistar-to-Wistar); Group Ⅱ: acute rejection (SD-to-Wistar); Group Ⅲ: thymus inoculation of SD rat LSA day 7 before transplantation. The observation of general situation and survival time, hepatocyte apoptosis and LAT expression in liver transplants were used to analyze immune state of animals in different groups. RESULTS: The general situation of group Ⅰ was very well after transplantation. Recipients of groupⅡ lost body weight progressively and all died within day 9 to day 13 post transplantation. As for group Ⅲ, the general situation of recipients was remarkably better than that in group Ⅱ. The positive cells of apoptosis in group Ⅲ detected by TUNEL were not significantly different from that in group Ⅰ, but was significantly lower than that in group Ⅱ. LAT was detected at any time in group Ⅱ with peak expression at day 5 and day 7 post transplantation. In contrast, LAT was not detected in any other groups. CONCLUSION: Intrathymic inoculation of LSA protects hepatocytes from apoptosis after liver allotransplantation.     

Novel Canine Tumour Suppressor Gene p53 Mutations in Cases of Skin and Mammary Neoplasms

Exons 4 to 8 of the tumour suppressor gene p53 were analysed in 25 skin and 25 mammary tumours of 50 dogs. A 1 bp deletion (ACCAC) was detected in codon 89 in exon 4 in a squamous cell carcinoma. A missense mutation CGCCAC (argininehistidine) was present in codon 162 in exon 5 in a mammary adenocarcinoma. Moreover, a silent mutation occurred in codon 103 (serine) of exon 4 in a mammary adenoma. The somatic nature of the three mutations was demonstrated.     

COMPUTED TOMOGRAPHIC QUANTIFICATION OF CANINE ADRENAL GLAND VOLUME AND ATTENUATION

We conducted a retrospective study in presumed normal dogs to determine the adrenal gland attenuation and volume values. Multidetector computer tomography (MDCT 16) analysis of the gland was carried out in 48 adult dogs without evidence of adrenal gland disease that underwent CT examination for acute spinal injuries. The mean nonenhanced attenuation value +/- SD of the left adrenal gland was 36.0 +/- 5.3 HU (range: 22.0-42.0 HU). The mean nonenhanced attenuation value +/- SD of the right gland was 34.3 +/- 7.0 HU (range: 20.4-48.6HU). The mean enhanced attenuation value +/- SD were: left gland 101.5 +/- 10.6HU (range: 86.8-128.0 HU), and right gland 97.4 +/- 12.4 HU (range: 58.9-123.6 HU). The mean CT volume +/- SD were: left gland was 0.60 cm3 (range: 0.20-0.95; SD 0.17), and right gland (0.55cm3, range: 0.22-1.01; SD 0.19). Attenuation values and volume data were related to age, weight, and gender, using ANOVA. There was no statistically significant difference between the left and right side or in adrenal measurements, because of body weight class effects. The animal effect was the most important source of variation for all adrenal measurements. Based on our study, CT is an effective method for assessing adrenal characteristics in the dog. Normative CT data are provided to allow estimation of normal adrenal gland size and volume.     

高温应激诱导奶牛乳腺细胞生长周期阻滞

实验通过台盼蓝染色、MTT、细胞流式等方法研究高温(40℃/41℃)对乳腺细胞活性、细胞周期及凋亡的影响。结果表明,高温可以抑制乳腺细胞的增殖,使细胞产生S、G2期阻滞,诱导乳腺细胞凋亡,并随高温强度的增加而作用增强。试验提示高温可诱导乳腺细胞周期阻滞,抑制细胞增殖。     

ULTRASONOGRAPHIC DESCRIPTION OF CANINE MASTITIS

Ultrasonographic images were acquired of the mammary glands of 40 bitches with physiologically lactating (n = 20) or inflamed glands (n = 20). Echogenicity, structure, homogeneity, thickness, and distinguishability of each tissue layer were assessed. Additionally, overall echogenicity was noted. In the normal lactating gland, different tissues could be differentiated easily. The parenchyma was, without exception, separated from adjacent tissues and was visible as medium echogenic tissue with a coarse-grained structure. The tissue always had some echogenic lines and anechoic areas and was slightly heterogeneous. The loss of distinct layering of the tissue was characteristic of an inflamed mammary gland and inflamed regions had reduced echogenicity. Additionally in five bitches with mastitis, the ultrasound examination was repeated five times for documentation of the progress of the illness and associated changes, supplemented with a color Doppler sonogram to assess changes in blood vessel density. Information from the examinations carried out via B-mode did not allow treatment success to be predicted. Two bitches with reduced blood vessel density centrally had a poor outcome whereas three bitches with increased blood vessel density had a good outcome. Thus, Doppler sonography might be a useful tool to obtain information of the prognosis in acute canine mastitis.     

Malignant pheochromocytoma presenting as a bradyarrhythmia in a dog

Objective: To describe a case of atrioventricular block and syncope secondary to systemic hypertension in a dog with malignant pheochromocytoma. Case Summary: A 13‐year‐old spayed female mixed‐breed dog presented following an acute onset of collapse. The dog was hypertensive and had paradoxical high‐grade second‐degree atrioventricular block with concurrent syncope. Initial emergent therapy included a nitroprusside infusion for the systemic hypertension and vagolytic therapy for the bradyarrhythmia. A left adrenal mass was seen on abdominal ultrasound and was further characterized by MRI. Following medical stabilization with phenoxybenzamine and propantheline, a left adrenalectomy was performed. Histological diagnosis was consistent with a malignant pheochromocytoma. Following tumor removal there has been no further hypertension, atrioventricular block or syncope. Unique information provided: This is the first documented case of a malignant pheochromocytoma presenting as syncope secondary to hypertension induced atrioventricular block.