The comparison of the collagen fiber contents and hepatic stellate cell distribution in male and female chicken livers

The difference in collagen fiber content, morphological properties and hepatic stellate cell (HSC) distribution was investigated in the liver of both sexes in chicken. Collagen fiber specimens were obtained by maceration treatment with NaOH solution. HSCs were detected using desmin‐specific immunohistochemistry. The ratio of liver weight to body weight was larger in the female than the male chickens. Collagen fiber content, the numerical density of HSCs and the percentage area displaying desmin immunopositivity were not different between the right and left lobes of the liver, in both male and female chickens. However, all of these parameters were larger in the males than the females. In the light microscopic observation, many HSCs in the male had large and elongated cytoplasmic processes. Conversely, HSCs with poorly developed cytoplasmic processes were frequently observed in females. Liver tissue is structurally stronger in male chickens than females and the activity and density of HSCs may be related to the collagen fiber content in chicken liver.     

Changes in collagen fiber content and hepatic stellate cell distribution in the liver of chick embryos and growing chickens

The content of collagen and the distribution of hepatic stellate cells (HSCs) were studied to elucidate the occurrence of sex‐dependent variations in the liver of developing embryos and growing chickens. Chick embryos from embryonic days (e) 12 to e20 and chicks at 1, 4 and 8 weeks were analyzed. Liver tissue was processed using NaOH maceration and freeze‐dried to obtain the collagen fiber specimens. HSCs were identified by double fluorescent immunohistochemistry for desmin and vimentin. There were no sex‐dependent variations in the percentage of collagen fiber per liver weight and HSC area during embryonic stages. However, the content of collagen fiber increased during embryonic development in both sexes. On the other hand, the area of HSCs significantly increased in growing males but did not show any change in females. Importantly, sex differences were observed in both collagen fiber content and HSC area in the liver at 8 weeks. These results indicate that the occurrence of collagen content variations takes place at 8 weeks in chicken liver, suggesting that a sex‐dependent hormone may play an important role on the collagen production of HSCs in the growing chicken liver.     

Cytokeratin‐positive folliculo‐stellate cells in chicken adenohypophysis

Folliculo‐stellate (FS) cells are non‐endocrine cells found in the adenohypophysis and are identified in many animals by the S100 protein marker. Although keratin is another FS marker in several animals, there is no information on localization of keratin in the avian adenohypophysis. In this study, localization of cytokeratin in chicken adenohypophyseal cells was investigated immunohistochemically. Basic cytokeratin (bCK)‐positive cells were arranged radially in the cell cords with their cytoplasmic processes reaching the basal lamina. The cell bodies encircled a follicle in the center of the cell cord. Furthermore, the bCK‐positive cells were also S100B‐positive. Growth hormone, prolactin, adrenocorticotrophic hormone, and luteinizing hormone β‐subunit did not co‐localize with the bCK‐positive cells. In addition, the bCK‐positive cells had a laminin‐positive area in their cytoplasm. Transmission electron microscopy observed agranular cells equipped with several microvilli that encircled a follicle. These results indicate that bCK‐positive cells in the chicken adenohypophysis may be a predominant FS cell population and produce laminin. It is suggested that they function as sustentacular cells to sustain the adjacent endocrine cells and the structure of the cell cords in the chicken adenohypophysis.     

Immunohistochemical and scanning electron microscopic comparison of the collagen network constructions between pig, goat and chicken livers

The distribution and three-dimensional architecture of collagen fibers were compared between pig, goat and chicken livers. Immunohistochemical staining revealed that collagen type I was identified in the interlobular connective tissue region and intralobular areas in pigs and goats. Type III collagen was also identified in the interlobular connective tissue region and intralobular sinusoidal walls. In the chicken liver, only the circumference region of the vessels was immunostained with collagen type I and III antibodies and the interlobular connective tissue wall could not be distinguished clearly. In the intralobular region, collagen type I antibody immunoreacted around the hepatic cells but collagen type III antibody immunoreacted weakly. In the NaOH macerated specimen, well-developed collagen bundles formed the prominent interlobular walls in pigs. In contrast, the wall in the goat liver comprised a thin layer of the bundles. In the chicken liver, there were no notable collagen septa between lobules. The intralobular collagen construction was quite different between the animals, indicating a fragile collagen fibril networks in pigs, a robust framework in goats and dense fabric-like septa in chickens. These results indicate that the distinct collagen frameworks may contribute to the histological strength of the livers in each of the animal species.     

Effects of Estradiol on Expression of Estrogen Receptor and Collagen mRNAs in Chick Skin

Skin thickness and strength differ between male and female chickens. This study aimed to clarify the effects of estradiol on the expression of estrogen receptors and collagen mRNA in chicken skin. Estradiol was administered to male chicks for 3 weeks, then cryosections of skin collected from the cervical, thoracic, dorsal, and pelvic limb regions were stained with hematoxylin and eosin, and dermal thickness was measured. Estrogen receptor and collagen mRNA expression was assessed using real-time RT-PCR, and collagen contents were determined. Estradiol did not alter dermal thickness or the collagen content of the skin from any tested region. Among the estrogen receptors, significantly more ESR1 mRNA was expressed in the thoracic skin of chicks administered with estradiol compared with vehicle (control), and in the thoracic skin compared with skin from other regions within each group. Estradiol did not affect ESR2, GPER, and COL1A1 mRNA expression. These results suggested that estradiol stimulates ESR1 expression in thoracic skin, but does not affect collagen synthesis in skin from any other region of male chicks.     

饲喂鸡肝对幼猫骨代谢的影响

选取50日龄左右健康短毛家猫20只,分为2组,每组10只,一组饲喂商品猫粮,另一组饲喂鸡肝,研究饲喂鸡肝对幼猫骨代谢的影响。结果表明:饲喂鸡肝可对幼猫的骨代谢产生明显影响,如显著降低血清钙(Ca)、骨钙素(BGP)和25OHD3含量;同时,血清碱性磷酸酶活性(AKP)和降钙素(CT)含量显著升高。鸡肝组幼猫在饲喂到50d时血清甲状旁腺激素(PTH)含量显著升高,但在饲喂到85d时显著降低。在饲喂到90d时,鸡肝组幼猫股骨长度极显著低于猫粮组(P<0.01)。     

Increased collagen III in culled chicken meat after feeding dietary wood charcoal and vinegar contributes to palatability and tenderness

We comprehensively evaluated meat quality in chickens fed a diet consisting of wood charcoal and vinegar (WCV) using food scientific and histological approaches. In culled hens, lipid and fatty acid in Musculus semimembranosus, cooking loss and sensory tests of whole thigh meat, and meat texture of breast meat were observed. In male broilers, cross section of M. semimembranosus was used for observations on muscle area, perimysium, non‐collagen total protein and total collagen content, and anti‐collagen I and III reactions. In frozen male broilers, conventional morphology of M. semimembranosus as well as chicken anti‐collagen III reaction to selected muscles of thigh meat and breast meat were compared between the control and WCV‐fed birds. Increased lipid and fatty acids, decreased cooking loss, high score in total evaluation for sensory test of thigh meat, and decreased meat texture values were observed for culled hens fed WCV. The higher values of muscle area, total collagen and collagen III were observed for broilers fed WCV. No perimysium collapse for M. semitendinosus or increased collagen III reactions of M. tensor fasciae latae, the flexor muscle group and M. pectoralis superficialis were observed for frozen muscles in the WCV group. These total results suggest that WCV produces palatable and tender meat by increasing collagen III.     

酶联免疫法测定鸡肝中氯霉素残留

运用酶联免疫检测试剂盒测定鸡肝中氯霉素的残留量.氯霉素标准液浓度在 0.025～2 μg/mL范围内线性良好,r=0.995 1,鸡肝中最低检测限为0.10 μg/kg.空白添加阳性样品浓度为0.1、0.5和1.0 μg/kg时,回收率分别为86.9%、89.1%和80.0%,板内变异系数<14.6%,板间变异系数<12.0%;本方法操作简单快捷、灵敏准确、方便实用,可作为鸡肝中氯霉素残留检测的快速筛选方法.     

Contribution of insulin to the ascorbate recycling system in the chicken liver

The effect of insulin on the ascorbate recycling system in the chicken liver was examined. First, insulin was injected subcutaneously into the chicken, and liver glutathione‐dependent dehydroascorbate reductase (GSH‐DHAR) activity was determined. Insulin increased liver GSH‐DHAR activity, but did not affect plasma and liver ascorbate concentration. Dehydroascorbate increased plasma and liver ascorbate levels, and liver GSH‐DHAR activity. However, distinct changes in plasma insulin level were not observed by dehydroascorbate injection. In addition, reduction of external dehydroascorbate in cultured chicken hepatocytes could not be observed in an insulin‐deprived culture, although the cells reduced external dehydroascorbate in a serum‐free culture with insulin. We concluded that insulin affects the ascorbate recycling system as an essential factor in the chicken liver.     

Oral ingestion of collagen peptide causes change in width of the perimysium of the chicken iliotibialis lateralis muscle

Skeletal muscle is mainly composed of myofibers and intramuscular connective tissue. Bundles composed of many myofibers, with each myofiber sheathed in connective tissue called the endomysium, are packed in the perimysium, which occupies the vast bulk of the intramuscular connective tissue. The perimysium is a major determination factor for muscle texture. Some studies have reported that collagen peptide (Col-Pep) ingestion improves the connective tissue architecture, such as the tendon and dermis. The present study evaluated the effects of Col-Pep ingestion on the chicken iliotibialis lateralis (ITL) muscle. Chicks were allocated to three groups: the 0.15% or 0.3% Col-Pep groups and a control group. Col-Pep was administered by mixing in with commercial food. On day 49, the ITL muscles were analyzed by morphological observation and the textural property test. The width of the perimysium in the 0.3% Col-Pep group was significantly larger than other two groups. Although scanning electron microscopic observations did not reveal any differences in the architecture of the endomysium, elastic improvement of the ITL muscle was observed as suggested by an increase of the width of perimysium and improved rheological properties. Our results indicate that ingestion of Col-Pep improves the textural property of ITL muscle of chickens by changing structure of the perimysium.     

Immunophenotypic analysis of the distribution of hepatic macrophages,lymphocytes and hepatic stellate cells in the adult rat liver

The liver consists of parenchymal hepatocytes and non-parenchymal cells. Non-parenchymal cells, Kupffer cells, hepatic stellate cells and cholangiocytes have crucial roles in liver homeostasis and liver pathology. To establish baseline data, this study investigated immunohistochemically the distribution of non-parenchymal cells in perivenular areas (PV), periportal areas (PP) and Glisson's sheath (GS) of adult rat liver. Liver tissues were collected from the left lateral lobe of rats. CD163-positive macrophages were seen along the sinusoid of PV and PP areas, indicating Kupffer cells. Double immunofluorescence showed, Kupffer cells partly co-expressed CD68 and MHC class II antigens in the liver. The numbers of Kupffer cells were significantly high in PP areas as compared with PV or GS areas. CD68-positive exudative macrophages were highly localized in PP and GS areas and a comparatively low PV area. MHC class II-positive dendritic cells (activated macrophages) were localized mainly in GS. Granzyme B-positive NK cells were mainly localized in the Glisson's sheath. CD3-positive T cells and CD20-positive B cells were distributed along the sinusoids of the PP and PV areas of hepatic lobules. Vimentin and glial fibrillary acidic protein (GFAP)-positive hepatic stellate cells were localized along sinusoids in the hepatic lobules of the liver. Cholangiocytes reacting to cytokeratin 19 were seen on interlobular bile ducts in Glisson's sheath of the liver. This study shows that heterogeneous macrophage populations, liver-resident lymphocytes and hepatic stellate cells localized in PP and PV areas or GS areas of the liver with cells specific patterns.     

Functional role of type VI collagen during early feather development of the chicken embryo in vitro

The regulatory function of type VI collagen during early feather development in embryonic chickens was investigated at the cellular and organ levels. Immunohistochemical studies of embryonic chicken skin showed that type VI collagen was distributed in spatial‐specific and temporal‐specific manners related to early feather development. To clarify the role of type VI collagen, we studied the feather development in intact, reconstituted and reconstituted gel skin cultures. When ethyl‐3,4‐dihydroxybenzoate (EDHB) was added to the medium of intact skin as an inhibitor of type VI collagen synthesis, the feather buds did not elongate and the number of neural cell adhesion molecule (NCAM)‐positive cells was reduced. However, the magnitudes of both suppressive effects of EDHB were reduced by the addition of liquid type VI collagen. Similar improvement was also observed in the reconstituted skin with liquid type VI collagen and in the reconstituted gel skin with solid type VI collagen at a low concentration. Moreover, type VI collagen promoted feather bud development in the absence of EDHB. However, a high concentration of solid type VI collagen in the reconstituted gel skin arrested the feather bud elongation, and antitype VI collagen antibodies caused feather buds to become longer and smaller in the reconstituted skin. At the cellular level, type VI collagen affected the proliferation, migration and NCAM expression of mesenchymal cells. These results suggest that type VI collagen regulates early feather development by controlling mesenchymal cell behavior.     

Gait deficits in liver disease: Hepatic encephalopathy and hepatic myelopathy

In this issue, the unusual clinical presentation of a horse diagnosed is described with severe liver cirrhosis and hepatic encephalopathy. The horse initially presented for thoracic and pelvic limb ataxia and weakness, and signs of forebrain disease were not apparent until later in the disease process. The typical pathology of central nervous system disease associated with liver disease is related to encephalopathy and forebrain disease; however, the spinal cord is occasionally also involved. Hepatic myelopathy is a rare syndrome usually associated with surgical or acquired portosystemic shunts, liver cirrhosis and/or portal hypertension in man. Where a gliopathy is the most prominent pathological feature seen in hepatic encephalopathy, in hepatic myelopathy the most remarkable feature is demyelination of the corticospinal tracts of the distal cervical and thoracic spinal cord with occasional axon loss. The clinical signs of hepatic myelopathy are spastic paresis/paralysis with normal sensory findings and preserved sphincter function. The prognosis for hepatic myelopathy is generally poor. In summary, in severe liver disease, motor deficits can occur secondary to the encephalopathy, but motor deficits can also occur as a result of spinal cord pathology such as seen in hepatic myelopathy. In examination of horses with myelopathies, liver disease as a cause of myelopathy should be included in our list of differentials.     

高效液相色谱-串联质谱法测定鸡肝中利巴韦林及其代谢物残留总量

建立了鸡肝组织中利巴韦林及其代谢物残留总量检测的高效液相色谱-串联质谱（HPLC-MS/MS）方法。鸡肝经磷酸酯酶水解、苯硼酸（PBA）固相萃取柱净化、C18色谱柱分离、电喷雾离子化（ESI+）和选择反应监测（SRM）方式采集,并以13C5-利巴韦林为同位素内标定量。该方法的检出限为0.5 ng/g;利巴韦林的测定在0.5 ng/mL～20 ng/mL 范围内线性关系良好,线性相关指数r为0.9997;在1.0 ng/g、2.0 ng/g和10 ng/g三个添加浓度的平均回收率为66.8%～110.3%,批内相对标准偏差6.8%～11.7%,批间相对标准偏差7.3%～10.1%。     

高效液相色谱法测定鸡肝组织中的氢溴酸常山酮残留

用高效液相色谱法测定鸡肝组织中常山酮的残留量.色谱柱为Kromasil C18(250 mm×4.6 HDM, 5 μm);流动相为乙腈-醋酸盐缓冲液(0.25 mol/L,pH 4.3)-水(5∶3∶12),检测波长243 nm,流速2.0 mL/min.该方法线性范围为0.10～5.00 mg/mL,r=0.999 9,平均回收率达到77.0%～80.3%,RSD<4.3%.该方法灵敏、准确、重现性好.     

QuEChERS-UPLC-MS/MS快速测定鸡肝中七种磺胺类药物残留

建立了一种通过QuEChERS方法和超高效液相色谱-串联质谱法快速测定鸡肝中七种磺胺类药物残留量的方法。样品以磺胺二甲氧嘧啶-D6为内标,经DisQuE萃取管提取,DisQuE净化管净化,浓缩后经超高效液相色谱分离,串联质谱测定。结果表明在12.5～125 ng/mL范围内各磺胺类药物均呈良好的线性关系,高中低浓度回收率在70%～115%,精密度小于15%。该方法快速、简便,适于鸡肝中七种磺胺类药物的批量测定。     

犬腹腔镜肝部分切除术对肝、肾功能的影响

选择8条健康杂种犬进行腹腔镜下肝部分切除,于麻醉前、术后4h及术后第1、3、5、7、9、11、13、15天采取静脉血进行肝肾功能指标监测.结果显示,天门冬氨酸氨基转移酶、丙氨酸氨基转移酶及碱性磷酸酶于术后第1天升到高峰；尿酸、尿素氮和肌酐于术后4h即达到高峰；白蛋白和总蛋白略微有所下降,于术后第1天降到最低；而r谷氨酰基转移酶术后变化不大.结果表明,犬腹腔镜肝部分切除术仅仅短期内对肝、肾功能产生影响,且肝、肾功能在术后一段时间内可恢复.     

鸡肝组织中拉沙洛西钠残留的高效液相色谱检测方法

建立了鸡肝组织中拉沙洛西钠残留的高效液相色谱检测方法.甲醇提取鸡肝组织样品中残留的拉沙洛西钠,硅胶柱净化,以磷酸盐缓冲液-乙腈-甲醇作为流动相,反相高效液相色谱-荧光检测法检测.方法平均回收率为82.1%,平均变异系数为7.75%,方法的检测限为0.02 mg/kg.     

鸡传染性贫血免疫对肝微粒体抗氧化酶活性的影响

目的是观察鸡传染性贫血多次免疫对肝微粒体中抗氧化酶活性的影响.对20只SPF鸡随机分为2组,每组10只,免疫组鸡用鸡传染性贫血弱毒苗免疫4次,每次间隔2周,对照组鸡注射同剂量的生理盐水.最后一次免疫后10d取肝脏制备微粒体,利用测试盒测定肝微粒体中的GSH-Px活性、SOD活性、CAT活性和MDA含量.结果与对照组相比,免疫组肝微粒体中GSH Px活性、SOD活性和CAT活性都显著提高(P＜0.05),MDA含量显著减低(P＜0.05).结论为鸡传染性贫血多次免疫可提高鸡体抗氧化能力.