Adrenomedullin (AM) and adrenomedullin binding protein (AM-BP) in the bovine mammary gland and milk: Effects of stage of lactation and experimental intramammary E. coli infection

Adrenomedullin (AM) has been characterized as an endogenous tissue survival factor and modulator of many inflammatory processes. Because of the increased susceptibility of the mammary gland to infection during the time surrounding parturition in the cow, we investigated how milk and tissue content of AM and its binding protein (AM-BP) might be affected by the stage of lactation and the udder health status. Milk and mammary biopsy samples were obtained from Holstein cows 21 days prior to and at various times after calving to represent the dry period and early and mid-stages of lactation. Additional cows received an intramammary challenge with Escherichia coli for immunohistochemical characterization of AM and AM-BP. Milk AM concentrations were relatively constant across the stages of lactation while AM-BP increased two-fold (P<0.04) between early and mid-lactation. Milk AM (P<0.04) and AM-BP (P<0.03) increased as somatic cell counts (SCCs) increased within a given stage of lactation. Tissue content of both (AM and AM-BP) were significantly affected by stage of lactation, lowest in the dry period and progressively increasing to peak at mid-lactation as well as increasing in association with higher levels of SCCs. Following E. coli challenge, AM increased in epithelial cells surrounding mammary alveoli presenting high levels of SCCs. The data suggest that AM and AM-BP are cooperatively regulated in the mammary gland during lactation; changes in localized tissue AM and AM-BP content reflect a dynamic regulation of these tissue factors in the bovine mammary gland consistent with their protective effects within inflamed tissue.     

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.     

Histology and histochemistry of the major salivary glands in the southern white-breasted hedgehog (Erinaceus concolor)

This study aimed to investigate the major salivary glands in the southern white-breasted hedgehog (Erinaceus concolor) histologically and histochemically. Five adult males were included in this study. The results showed that anatomically the shape of the parotid, submandibular and sublingual glands of the Erinaceus concolor was respectively almost pear, elliptical to pyramidal and oval. Histologically, the parotid gland had serous acini and secretory cells showed negative reaction to alcian blue (AB) (pH = 2.5) and negative response to aldehyde fuchsin (AF), methylene blue (MB) and PAS stains. The submandibular gland was a mixed gland of mucous and serous acini. The mucous acini were strongly positive for PAS, AB, MB and AF. However, serous acini were week for PAS and negative against AB, MB and AF stains. The sublingual gland was purely composed of mucous acini. The mucous acini of the sublingual gland were strongly positives for PAS, AB and MB methods. While their reaction to the AF staining was negative. In conclusion, the histological and histochemical observations of the major salivary glands of the southern white-breasted hedgehog (E. concolor) indicated that these glands shown similarities and some special different histochemical features as compared to other mammalian species.     

Somatic cell and innate immune responses in mammary glands of lactating cows to intramammary infusion of Bifidobacterium breve at pre-drying off period

Intramammary infusion of Bifidobacterium breve (B. breve)-induced somatic cell (SC) counts, chemiluminescent response (CL), lactoferrin (LF) concentrations and mastitis-causing pathogens from quarters with subclinical mastitis were measured to evaluate innate immune response of mammary glands in dairy cows at 3 to 4 weeks before drying off. SC counts in 7 quarters of 7 control cows and 5 quarters of 6 cows with mastitis increased markedly on day 1 and SC values in control cows were significantly (P<0.05) increased and returned to pre-infusion levels on day 5 after B. breve-infusion. CL values in both groups increased markedly on day 1 and then decreased after B. breve-infusion; however, CL values in cows with mastitis did not return to normal levels on day 5 and at postpartum. The CL values were highly correlated with their SC counts in milk from both groups. LF concentrations increased toward day 3 after B. breve-infusion and were higher in cows with mastitis. B. breve-infusion eliminated 16.6% (1/6) of pathogens from 6 quarters with chronic subclinical mastitis. B. breve-induced SC responses in quarters from 3 cows with mastitis showed characteristic patterns of recovery, persistent and new infections. B. breve-induced SC counts in quarters from the cows in the pre-drying off were lower (25.7–70.6%) than those of the cows in mid-lactation. The intrinsic innate immune response in cows on pre-drying off may be decreased and appears to be insufficient to eliminate pathogens from mammary gland in the pre-drying off.     

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。     

Malignant pinealoma observed in the deep cerebral parenchyma of a male Wistar rat

This report describes a case of spontaneous malignant pinealoma in a 90-week-old male Wistar rat. The tumor mass occurred in the deep cerebral parenchyma and no intact pineal gland was observed in the area between the posterior-dorsal median line of the cerebrum and the cerebellum. The tumor was characterized by a large nodular proliferation occupying the central area of the brain, extending from the dorsal surface to the base of the brain, corresponding to the thalamus. The tumor cells had round to irregular oblong nuclei approximately 5–17 μm in diameter and showed faintly or moderately eosinophilic cytoplasm and indistinct cell boundaries. Immunohistochemically, the tumor cells were positive for synaptophysin and partially positive for neuron-specific enolase (NSE). The tumor showed malignant features including cellular pleomorphism, high mitotic index, necrotic foci, and invasive and extensive growth and was, therefore, diagnosed as an extremely rare malignant pinealoma in the deep cerebral parenchyma.     

蚓激酶基因乳腺表达载体构建及在牛乳腺中的瞬时表达

在稀有密码子的改造基础上,人工合成了849 bp的蚓激酶F-Ⅲ-1全长cDNA序列,并以其为目的基因,构建了以山羊β-酪蛋白启动子为上游调控序列的乳腺组织特异性表达载体pBLK和pLN-bCP-LK。2种质粒分别以200,500,1 000,2 000μg的剂量梯度注入泌乳期黑白花奶牛乳腺,以纤维蛋白平板溶圈法(FAPA)检测该基因表达后奶样的纤溶活性。结果表明:注射后3~78 h,奶样有明显纤溶活性,其中6~16 h活性最高。500μg pBLK质粒与其他3个剂量溶圈直径之间差异显著(P<0.05),而其他3个剂量之间差异不显著;pLN-bCP-LK质粒的各剂量溶圈直径之间差异均不显著,且2种质粒的同剂量溶圈直径之间差异也不显著。以上结果表明,改造后的蚓激酶基因可以在泌乳期黑白花奶牛乳腺中实现瞬时表达,以500μg的注射剂量表达效果最佳。