Calpain活性的调节及其在细胞转化和迁移中的作用

钙蛋白酶(Calpain)是一类钙依赖性的高度保守的蛋白水解酶.近年来,有关 Calpain三维晶体结构以及Calpain基因的研究取得了很大的进展.研究发现,Calpain参与细胞骨架蛋白重整、细胞转化和迁移,加快细胞循环.此外,Calpain参与的细胞转化和迁移可能与癌症有一定的关系.因此,深入研究Calpain将有助于进一步认识癌症发生的病理学机制.     

Interactions between metabolic status, pre-breeding protein supplementation, uterine pH, and embrionic mortality in ewes: Preliminary observations

The effect of prebreeding short-term protein supplementation level (PL) and body condition (BC) on fertility rate (FERT,%), uterine pH and embryonic mortality (EMORT,%) in sheep, was evaluated. Multiparous Rambouillet ewes at low BC (LC; n = 6, 62.7 ± 1.7 kg) or high BC (HC; n = 6, 71.9 ± 1.7 kg), received. within BC, one of two levels of ruminally undegradable protein: low (LP, 14 g/ewe per day) and high (HP, 30 g/ewe per day). Once the animals were euthanized, corpus luteum number (CLN), as an indicator of ovulation rate, was registered and uterine horns were irrigated to recover embryonic tissue plus associated membranes as well as to measure uterine pH (UpH). While EMORT-1 considered a nonadjusted relationship between the number of embryos and CLN, EMORT-2 considered an analysis of covariance using CLN as the covariate. The HP-supplemented ewes had the lowest FERT (p=0.06; 100% vs 50%) and the highest EMORT (EMORT-1, 16.6% vs 53.8%, p = 0.08; EMORT-2, 52.0% vs 14.5%, p = 0.07) when compared to the LP-supplemented ewes. Neither BC nor PL affected CLN, CL weight or P₄ release (p > 0.10). While the lowest UpH (p = 0.04) was observed in the HP-supplemented ewes, this group also showed the lowest fertility and the highest embryonic mortality.     

Combined treatment with oestradiol benzoate,d‐cloprostenol and oxytocin permits cervical dilation and nonsurgical embryo recovery in ewes

This study examined the feasibility of transcervical embryo recovery after the hormonal treatment to induce cervical dilation, following the 7‐day oestrous synchronization protocol in multiparous Santa Inês ewes. A total of 23 cyclic ewes received two doses of 37.5 μg of d‐cloprostenol by latero‐vulvar route 7 days apart. After the second injection of d‐cloprostenol, the ewes were checked for oestrus (every 12 hr) and then mated by fertile rams throughout the oestrous period. All ewes received 37.5 μg of d‐cloprostenol (latero‐vulvar) and 1 mg of oestradiol benzoate by either intramuscular (EBim group; n = 12) or intravaginal (EBivg group; n = 11) route 16 hr before embryo flushing. Twenty minutes before the flushing, 50 IU of oxytocin were administered intravenously. The oestrous response (i.e., the percentage of ewes that showed signs of oestrous behaviour after the second d‐cloprostenol injection) was 91.3% (21/23). The proportion of successfully penetrated ewes (81.8% compared with 80.0%), the mean duration of embryo flushing (24.7 ± 2.0 min compared 26.2 ± 1.9 min), the flushing fluid recovery rate (94.8 ± 1.3% compared with 91.0 ± 2.9%) and the average number of structures recovered per ewe (0.5 ± 0.4 compared with 0.8 ± 0.4) did not vary (p > 0.05) between the EBim and EBivg groups. Viable embryos were recovered from 41.2% (7/17) of successfully penetrated ewes. It can be concluded that nonsurgical (i.e., transcervical) embryo collection can be performed in oestrous‐synchronized Santa Inês ewes pretreated with d‐cloprostenol, oxytocin and oestradiol benzoate, with the latter hormone administered by either the intramuscular or intravaginal route.     

Effects of summer heat stress on physiological variables,ovulation and progesterone secretion in Pelibuey ewes under natural outdoor conditions in an arid region

Ten non‐lactating multiparous Pelibuey breed ewes were housed in a corral to evaluate the effects of summer thermal stress on physiologic variables, estrous behavior, ovulation and corpus luteum functionality under natural conditions of an arid region. In summer and autumn, daily estrous detection with a ram fitted with an apron and blood sample collections were performed during two natural estrous cycles. An environment of heat stress was detected in summer and thermoneutral in autumn. Rectal temperature and respiratory frequency were greater (P < 0.01) in summer than in autumn during the morning and afternoon. Season did not affect (P > 0.05) live weight, body condition, length of estrous cycle or percentage of ewes in estrous and ovulating. Compared with autumn, serum progesterone concentrations in summer decreased (P < 0.05) between days 8 and 14 of the estrous cycle. It is concluded that under outdoor conditions of arid regions, while estrous and ovulatory activities of Pelibuey ewes were not affected by summer thermal stress, the corpus luteum functionality was decreased.     

2种紫花苜蓿氮代谢及其关键酶(NR、GS)活性差异研究

为探究不同紫花苜蓿氮代谢差异及氮代谢差异的生理机制,采用盆栽砂培法,通过对2个具有典型特征的紫花苜蓿品种(LW6010和陇东苜蓿)在2个氮素水平及3个生育时期下其氮代谢产物及氮代谢酶活性差异进行研究,探讨氮代谢产物与氮代谢酶活性的相互关系。结果表明:LW6010和陇东苜蓿在N210水平下其干物质重、氮含量、氮积累量及硝酸还原酶(NR)、谷氨酰胺合成酶(GS)活性均高于N21。在2个氮水平下,LW6010的干物质重、NR和GS活性均显著大于陇东苜蓿,而LW6010的氮含量显著低于陇东苜蓿。其中,LW6010的NR活性比陇东苜蓿在N210和N21下分别高24%和15%(苗期)、31%和40%(现蕾期)、20%和11%(盛花期);GS活性分别高30%和29%(苗期)、33%和20%(现蕾期)、7%和14%(盛花期)。在苗期和盛花期时,LW6010的氮积累量显著大于陇东苜蓿,而在现蕾期差异不显著。LW6010的氮积累量在N210下各生育期分别比陇东苜蓿高25%,3%和16%,而在N21下LW6010的氮积累量在苗期和盛花期下比陇东苜蓿高45%和12%。同时,紫花苜蓿干物质重、氮积累量、NR活性与GS活性呈极显著正相关关系。综上分析,紫花苜蓿的不同品种在不同氮素水平下其氮代谢能力不同,不同紫花苜蓿品种间氮代谢具有差异;LW6010较陇东苜蓿具有较好的氮代谢能力;NR和GS活性可为不同紫花苜蓿氮代谢差异评价提供参考。     

Death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer and their progeny: a result of nationwide survey in Japan

To obtain the data concerning death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer (SCNT) and their progeny produced by Japanese institutions, a nationwide survey was carried out in July-August, 2006. As a result, lifetime data concerning 482 SCNT cattle (97.5% of cattle produced in the country at that time) and 202 progeny of SCNT cattle were accumulated and the death loss of these cattle was analyzed. Although 1/3 of delivered SCNT calves died during the perinatal period due to stillbirth and neonatal death, incidence of death loss due to diseases in SCNT cattle surviving more than 200 days after birth seems to be the same as these in conventionally bred cattle. In contrast, progeny of SCNT cattle showed the same level in death loss as observed in conventionally bred cattle throughout their lifetime. These results suggest that robust health would be expected in SCNT cattle surviving to adulthood and their progeny.     

Faecal thiaminase, plasma lactate and pyruvate concentrations and erythrocyte transketolase activity changes in apparently normal replacement ewes after the initiation to the pasture

A study was made to investigate faecal thiaminase and the thiamine-related biochemical changes in apparently normal replacement ewes with a feed change, after the initiation without adaptation to the new pasture. Twenty-four female ewes were divided into two groups. Group A was managed in a system based on pasture and was compared with group B system based on a diet of concentrate and straw until moving to pasture 9 weeks after. Blood samples for lactate, pyruvate and erythrocyte transketolase activity determinations and faeces for thiaminase estimation were evaluated chronologically. At the end of a 126 days experimental period, live weights of groups were similar. We confirmed that clinically normal sheep may have thiaminase activity in the faeces and concluded that the thiaminase release increased during the diet changes, from concentrate to pasture, and that their continued excretion could develop some degree of thiamine deficiency.     

Diurnal and oviposition-related changes in heart rate, body temperature and locomotor activity of laying hens

Diurnal and oviposition patterns of heart rate (HR), deep body temperature (BT) and locomotor activity (LA) in conscious and unrestrained Rhode Island Red hens were studied by a radiotelemetry system. Behavioral observations were also made on diurnal changes and during the pre‐ and post‐laying period. Heart rate, BT and LA showed characteristic diurnal changes synchronized with a photoperiod of 15 h light and 9 h dark. In the light period, HR, BT, and LA levels were significantly higher than in the dark period (P < 0.001). Furthermore, the highest levels of these parameters were recorded just after they were fed (08.30 hours), while the lowest level was measured after lights‐off and remained stable throughout the dark period. Behavioral observations indicated that during the light period the hens spent most of their time in very active movement, exhibiting various behavioral patterns. However, in the dark period the hens spent almost all their time resting. The present results suggest that performing various behavioral activities cause heat generated by muscle exertion, which plays a significant role in daily HR, BT, and LA in laying hens. However, during the 60 min before and after oviposition, LA appeared to have increased steadily toward the moment of laying, and then regressed gradually in the post‐laying period to a level significantly lower than in the pre‐laying period (P < 0.001). Furthermore, the pre‐laying behavior of hens indicated extreme restlessness and more activity, whereas the post‐laying period is characterized by less activity and increased relaxation. Consequently, laying behavior has a profound but transitory effect on HR and BT, suggesting that oviposition was probably associated with intense LA.     

Effects of forage source and particle size on chewing activity,ruminal pH,and saliva secretion in lactating Holstein cows

The present study investigated the effects of dietary forage source (quality) and particle size on chewing activity, saliva secretion, and ruminal pH. Twelve multiparous lactating Holstein cows, four of which were ruminally cannulated, were used in a replicated 4 × 4 Latin square experimental design with a 2 × 2 factorial arrangement of treatments. Cows fed wild‐rye hay diets had longer daily eating times than cows fed oaten hay diets. Treatments had no effect on ruminating time; therefore, resting time varied inversely to eating time. Neither the rate nor the amount of saliva secretion while eating, ruminating, or resting was affected by diet, resulting in similar total daily saliva secretions across treatments (231 L/day). Total volatile fatty acids (VFAs) in the ruminal fluid from animals fed oaten hay diets were higher than those from animals fed wild‐rye hay diets; further, VFAs increased with decreasing forage particle size (FPS). Consistent with elevated VFA concentrations, reducing FPS and including oaten hay in the diet decreased mean ruminal pH and increased the daily time of ruminal pH under 5.8. Results of this study suggest that forage source and particle size affect ruminal pH might be via variations in VFA production rather than increased salivary recycling of buffering substrates.     

牛磺酸对肉仔鸡生产性能、养分利用率及脂肪酶活性的影响

本研究旨在探讨牛磺酸对肉仔鸡生产性能、养分利用率及脂肪酶活性的影响。选用240只1日龄AA肉仔鸡,随机分为5组,每组3个重复。对照组饲喂玉米-豆粕型基础日粮,试验组在基础日粮中添加0.05%,0.10%,0.15%和0.20%的牛磺酸,试验期42 d。结果表明:日粮中添加0.10%牛磺酸,能够显著提高肉仔鸡平均日增重和饲料转化率(P<0.05),显著提高18~21日龄粗蛋白利用率(P<0.05);0.15%水平的牛磺酸能够显著提高18~21日龄肉仔鸡的表观代谢能和粗脂肪利用率(P<0.05)及21日龄胰脏和小肠内容物脂肪酶活性(P<0.05);添加牛磺酸对39~42日龄肉仔鸡的养分利用率及42日龄胰脏和小肠内容物脂肪酶活性均无显著影响(P>0.05)。说明牛磺酸对肉仔鸡促生长作用与提高养分利用率有关,且粗脂肪利用率的提高与胰脏和小肠内容物脂肪酶活性提高有关。     

微粒蒙脱石对猪组织铅水平、红细胞生成和肝脏ALA-D酶活性及铅诱导的脂质过氧化的影响

选60头体质量约33kg的“杜长大”杂交猪，随机分成2组，每组3个重复，一组饲以基础日粮＋10mg/kg铅＋0.5％微粒蒙脱石，另一组饲以基础日粮＋10mg／kg铅作为对照。研究结果表明：添加微粒蒙脱石组与对照组相比，明显降低了猪全血、脑、肝、肾、骨和毛等组织中铅含量：通过血细胞计数、血红素和血球容积测定表明，添加微粒蒙脱石组的红细胞生成显著增高，且肝脏中ALA—D酶活性显著提高。铅对肝脏的损伤作用明显。表现为丙二醛（MDA）含量显著升高（17．08％），过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPx）含量分别降低85．73％、52．17％、47．56％，而在日粮中添加微粒蒙脱石可以显著改善铅诱导的上述损伤。结果提示，微粒蒙脱石对铅中毒有潜在的治疗作用。     

地塞米松、油酸、乳酸对体外培养脂肪细胞HSL活性的影响

为了研究地塞米松、油酸、乳酸对体外培养的脂肪细胞激素敏感脂肪酶（HSL）活性的影响,以体外培养生长良好脂肪细胞为实验模型,培养介质中分别添加0、10、20、40、80、160mg／I。的地塞米松,0、0．5、1．0、1．5、2．0、2．5mmol／L的油酸,0、10、20、30、40、50mg／L的乳酸,培养24h后提取总蛋白,每个处理3个重复,分别采用脂肪酶测定试剂盒检测HSL活性。结果表明,添加乳酸、油酸可抑制HSL的活性,随添加浓度增加,其抑制作用增强;地塞米松对HSL活性有促进作用,且存在剂量依赖性。     

全混合日粮中饲草长度对奶牛采食行为、瘤胃发酵和日粮消化率的影响

[目的]本研究旨在探讨奶牛TMR中,粗饲料羊草长度对奶牛采食行为、瘤胃发酵和日粮消化率的影响.［方法］试验动物为3头体重496士23 kg,安装有永久性瘤胃瘘管的健康中国荷斯坦奶牛.采用3×3拉丁方设计.用36% NDF不同羊草长度（9 cm、5 cm和1 cm）的三种全混合日粮饲喂奶牛.［结果］:①奶牛DMI和NDFI不受羊草长度影响;采食、反刍和总咀嚼时间随着羊草长度的减小显著减少.②羊草长度为1 cm时瘤胃pH显著低于9 cm和5 cm组;瘤胃内NH3-N浓度在5 cm组最高,与1 cm组差异明显;随着羊草长度减小,乙酸浓度和乙酸与丙酸的比例显著降低,丙酸浓度升高,总VFA和丁酸浓度不受羊草长度影响.③日粮中DM、CP、NDF和ADF消化率在羊草长度为5 cm时显著高于其他两组.表明饲草长度的减小会缩短采食、反刍和咀嚼时间,但过短会使瘤胃pH显著降低;适当长度可提高日粮营养成分的消化率.［结论］综合评价本试验结果以羊草长度为5 cm更适合全混合日粮配制.     

阿坝地区狼毒内生放线菌多样性及抗菌活性

通过可培养方法以四川省阿坝地区药用植物狼毒为研究对象,探究其内生放线菌多样性及抗菌活性,为开发新型生物活性物质提供依据.结果如下,1)多种因素决定了狼毒内生放线菌分离数量,其中土壤有机质与其相关系数为0.86,在0.05水平相关性显著.2)高氏一号培养基分离52株内生放线菌,占总数30.4%;植株不同部位分离内生放线菌结果为根部>茎部>叶部>花部.3)16S rDNA-RFLP将供试菌株分成11个遗传群,代表菌株系统发育分析表明,供试菌株以链霉菌属为主,其余菌株包括诺卡氏菌属,北里孢菌属,克里布所菌属.4)对代表菌株NRPS,PKS基因分析共获得4个PKS基因和2个NRPS基因;大部分代表菌株对3种病原真菌有抗菌活性,菌株SCAUEⅢD11-1效果最好.综上,狼毒内生放线菌分离结果受多因素影响,具有较丰富的多样性;功能基因筛选及抗菌活性结果揭示出阿坝地区狼毒内生放线菌具有潜在的运用价值.     

Effects of physically effective fiber in diets based on rice straw and cassava pulp on chewing activity,ruminal fermentation,milk production,and digestibility in dairy cows

The objective of this study was to evaluate the effects of physically effective neutral detergent fiber (peNDF) in rice straw and cassava pulp diets on the chewing activity, ruminal fermentation, milk production, and digestibility in low‐producing dairy cows. Eight lactating Holstein crossbred cows were randomly divided into two groups in a crossover design with two dietary treatments over two 21‐day periods. The difference in peNDF contents was achieved by balancing the ratio of rice straw, cassava pulp, and concentrate. The high cassava pulp diet had 15.5% peNDF and the low cassava pulp diet had 20.2% peNDF. The results showed that feeding the low cassava pulp diet increased the concentrations of milk protein, milk fat, solids‐not‐fat, and total solids in the milk. In contrast, the dry matter intake, milk yield, lactose content, chewing activity, nutrient digestibility, ruminal fermentation, and pH were not affected by treatments.     

Effects of gelatin prepared from calf bones rich in phosphorus on broiler performance,bone characteristics and digestive enzymes activity

1. Gelatin prepared from calf bones (GCB) is a novel source of high-quality protein and phosphorus. Its inclusion in broiler chicken diets may improve bone strength, plasma and digestive alkaline phosphatase activity (ALP), phosphorus digestibility and performance of broilers. Therefore, di-calcium phosphate in a corn-soy control diet was replaced with 12, 24, and 36 g/kg of GCB in a completely randomised design with four treatments of six replicates and 10 chicks in each pen. The trial lasted from 1 to 42 d of age.

2. Body weight and feed intake were measured weekly. Plasma calcium and phosphorus concentration along with plasma and digestive ALP were assayed throughout the trial. Trypsin, α-amylase, lipase and total protease activity were assayed at 14 and 28 d of age. Tibia ash, calcium and phosphorus content and breaking strength were measured at 14, 28 and 42 d of age. Phosphorus digestibility was measured at 36 d of age.

3. Body weight and feed intake showed no significant differences between controls and diets containing 12 and 36 g/kg GCB. Tibia ash and tibia length were increased by supplementation of GCB (P ≤ 0.001). Tibia calcium and phosphorus content were increased by GCB inclusion at 14 d of age (P ≤ 0.001). Digestive alkaline phosphatase activity was increased and trypsin activity was reduced by inclusion of GCB (P ≤ 0.001; P ≤ 0.004). α-amylase activity decreased by inclusion of 12 and 24 g/kg GCB, whereas an increase in α-amylase activity was observed by inclusion of 36 g/kg GCB (P ≤ 0.001). Supplementation of diets with GCB increased phosphorus digestibility (P ≤ 0.01) and suppressed ileum growth during the experimental period.

4. Results of the current study showed that phosphorus from gelatin can greatly improve broiler bone characteristics and phosphorus digestibility and complete elimination of inorganic phosphate sources from broiler diets is feasible with inclusion of 36 g/kg high phosphorus gelatin.     

Effects of toll-like receptor-7 agonists on feeding behaviour,voluntary activity,cloacal temperature and crop emptying in chicks

ABSTRACT

1. The purpose of the present study was to determine if an intraperitoneal injection of two toll-like receptor-7 (TLR7) agonists, imiquimod and resiquimod, affect feed intake, voluntary activity, cloacal temperature, crop-emptying rate, plasma corticosterone (CORT) and glucose concentrations, and splenic gene expression of cytokines in chicks (Gallus gallus).

2. Although intraperitoneal injection of 100 µg imiquimod significantly increased splenic gene expression of interleukin-1β (IL-1β), IL-6, IL-8, and interferon-γ (IFN-γ), it did not affect feed intake, voluntary activity, cloacal temperature, crop-emptying rate or plasma constituents.

3. Intraperitoneal injection of 100 µg resiquimod significantly decreased feed intake, voluntary activity, cloacal temperature, crop-emptying rate and increased plasma corticosterone concentrations.

4. Intraperitoneal injection of resiquimod significantly increased splenic gene expression of IL-1β, IL-6, IL-8, IFN-γ, and tumour necrosis factor-like cytokine 1A.

5. The results showed that activation of TLR7 is associated with anorexia, hypoactivity, hypothermia, disturbance of feed passage in the digestive tract and the response to stress in chicks.     

Relationships among vasculature,mitotic activity,and endothelial nitric oxide synthase (eNOS) in bovine antral follicles of the first follicular wave

To determine the relationships among vasculature, mitotic activity, and expression of endothelial nitric oxide synthase (eNOS) of antral follicles in Bos indicus, bovine ovaries were obtained on day 6 of the estrous cycle from 10 crossbred (Brahman to Thai native cows) after a synchronized estrus with prostaglandin F₂α analogue. Ovaries were fixed, paraffin-embedded, and used for immunofluorescence detection of factor VIII (a marker of endothelial cells). Immunostaining of eNOS and proliferating cell nuclear antigen (PCNA) were performed with specific monoclonal antibodies. Vasculature and positive staining of eNOS and PCNA were quantitatively evaluated with the image analysis. Follicles were classified by size (small, medium, and large) and by structure as healthy and atretic follicles (n = 82). The expression of factor VIII and eNOS were detected greater in the blood vessels of the theca layers of the healthy follicles than those in atretic follicles. The labeling indices (LIs) in granulosa and theca cells were greater (P < 0.05) in the healthy small and medium follicles than in the healthy large follicles. Vasculature, capillary area density, and capillary number density were positively correlated with eNOS expression and the LIs of granulosa and theca cells but were negatively correlated with the healthy follicle size. During the growing phase of antral follicle in Bos indicus, relationships among vasculature, mitotic activity, and eNOS were observed predominantly in healthy antral follicles. Thus, these data highlight the importance of vasculature, cell proliferation, and eNOS expression of growing and atretic follicles in the first follicular wave.     

不同种植年限条件下黄花蒿根际土壤微生物生物量、酶活性及真菌群落组成

试验采集未种植、种植1年、3年和5年的黄花蒿根际土壤,采用常规分析和Illumina MiSeq高通量测序技术,研究了土壤微生物生物量、酶活性及真菌群落组成。结果表明,在人工种植黄花蒿的土壤中,微生物生物量碳氮减少,碳氮比例改变;脱氢酶、脲酶和蔗糖酶活性降低,酸性磷酸酶活性增强;说明黄花蒿释放的化感物质选择性抑制了土壤微生物生长、繁殖和代谢。在不同种植年限的土壤中,主成分分析显示代表不同种植年限土壤真菌群落的点在坐标图中分布距离较远,表明它们的群落组成发生了显著变化(P<0.05)。此外,子囊菌门占土壤真菌的66.10%~95.28%,黄花蒿种植时间影响真菌门类和优势真菌的丰富度。在前20种优势真菌中,有14种共存于不同种植年限的土壤中,每种土壤中存在1~3种独有真菌,说明土壤是决定真菌种群组成的主导因素,又因种植黄花蒿而改变。在栽培1~5年的黄花蒿土壤中,优势菌株中出现蒿属的常见病菌——蒿白粉菌和艾菊柄锈菌,提高相应病害的发生风险。