牦牛肠源Lactobacillus acidophilus L3对其肠道分泌型免疫球蛋白A及免疫相关因子的影响

为了研究牦牛肠源Lactobacillus acidophilus L3(嗜酸乳杆菌)对其肠道分泌型免疫球蛋白A(SIg A)及免疫相关因子的影响,将10头健康牦牛(2~2.5岁)随机分为2组,分别为益生菌组和空白对照组,益生菌组在饲料中添加2×109CFU·kg-1L.acidophilus L3,饲喂28 d后取其小肠样品,ELISA检测试验组和对照组十二指肠、空肠、回肠中SIg A、白细胞介素-2(IL-2)、IL-4、IL-6、干扰素γ(IFN-γ)的含量。研究结果表明,L.acidophilus L3可有效增加试验组牦牛肠道SIg A的分泌量(P<0.05),组内比较发现SIg A在回肠含量最高,其次为空肠和十二指肠。IL-2、IL-4、IL-6在试验组小肠中的表达量较对照组显著增加(P<0.05),试验组小肠IFN-γ的表达量较对照组降低(P<0.05)。证实L.acidophilus L3可提高牦牛肠道的黏膜免疫功能。     

鹌鹑、红嘴鸥、黑翅长脚鹬胚胎期 小肠黏膜表面形态结构

应用扫描电镜观察了鹌鹑（Coturnix coturnix）、红嘴鸥（Larus ridibundus）、黑翅长脚鹬（Himantopus himantopus）胚胎期小肠黏膜的形态变化。鹌鹑与红嘴鸥胚胎发育过程中,小肠黏膜形态结构可分为3个阶段。第一阶段为鹌鹑卵孵育第10 ~ 11天、红嘴鸥卵孵育第13 ~ 14天（相当于胚胎发育60%的阶段）,小肠黏膜为山脊状纵行皱襞;第二阶段为鹌鹑卵孵育第12 ~ 13天、红嘴鸥卵孵育第15 ~ 16天（相当于胚胎发育70%的阶段）,小肠黏膜为“W”形板状皱襞;第三阶段为鹌鹑卵孵育第14 ~ 17天、红嘴鸥卵孵育第17 ~ 22天（相当于胚胎发育到80% ~ 100%阶段）,小肠黏膜为指状绒毛。黑翅长脚鹬卵孵育第10天（相当于胚胎发育到60%的阶段）,小肠黏膜为山脊状纵行皱襞;从卵孵育第12天（相当于胚胎发育到70%的阶段）一直到孵出小肠黏膜均为“W”形板状皱襞。初步判断3种鸟小肠黏膜形态发生这种有规律的变化可能是鸟类对其祖先系统发育的重演。     

肠黏膜更新的源泉——小肠干细胞

随着细胞分选和基因标记技术逐渐成熟,干细胞研究由原来的干细胞移植雏形—骨髓移植研究逐步发展成为一个新兴的学科。作为机体内细胞更新最快的组织器官之一——小肠,其干细胞的研究日益受到人们的关注。由于小肠黏膜单层上皮细胞的简单结构和特有的昼夜更新节律,小肠干细胞成为继造血干细胞和皮肤干细胞之后新的研究热点。同时,由于部分肠道疾病导致的黏膜破坏,肠功能难以自我代偿,小肠干细胞移植为肠黏膜的再生和肠功能的恢复提供了新的前景。     

超微七味白术散对菌群失调腹泻小鼠小肠黏膜的影响

目的 探讨超微七味白术散对抗生素相关菌群失调腹泻的疗效机制.方法 采用灌胃头孢拉啶和硫酸庆大霉素混合抗生素进行造模,然后分别灌胃七味白术散的传统汤药全量和超微汤药1/2剂量进行治疗,正常组和模型组分别灌胃等量的无菌水.结果 抗生素造模后,小鼠体重的变化量和变化率都较正常组小,差异无统计学意义(P＞0.05),小鼠的小肠黏膜组织发生绒毛脱落、炎症细胞浸润、固有膜毛细血管扩张等病理变化;传统汤药治疗组和1/2剂量超微汤药治疗组小鼠体重的变化量和变化率都比正常组、模型组大,且1/2剂量超微汤药组的与正常组的差异有统计学意义(P＜0.05),该两组小鼠的肠道组织病理情况有所改善,1/2剂量超微汤药治疗组小鼠的肠道组织恢复情况最好,传统汤药治疗组次之,模型组也稍有改善.结论 1/2剂量超微七味白术散对恢复菌群失调腹泻小鼠的小肠黏膜的结构具有较好的修复作用,对小肠吸收功能具有促进作用.     

牦牛ZP3基因的克隆及蛋白质结构的预测

本研究对牦牛ZP3基因的编码区进行了克隆,在此基础上对ZP3蛋白的分子结构预测,为研究牦牛受精生物学提供基础。根据GenBank中普通牛的ZP3核苷酸序列设计特异性引物,以牦牛卵巢组织总RNA为模板,通过RT-PCR技术扩增牦牛ZP3基因cDNA序列(GenBank登录号为GQ856646),利用DNAMAN生物软件进行核苷酸和氨基酸序列分析、蛋白质专家系统ExPASy进行ZP3蛋白质分子结构预测。结果表明,扩增出的牦牛ZP3基因编码序列长1 266 bp,编码421个氨基酸。牦牛与牛、猪、狗、人、鼠和鸡ZP3基因核苷酸相应序列的同源性分别为98.42%、96.73%、79.67%、78.71%、69.15%和56.61%,氨基酸同源性分别为98.10%、83.85%、74.24%、70.26%、62.62%和46.12%,符合物种进化规律。预测的ZP3蛋白二级和三级结构显示它是一个具有22个氨基酸信号肽的亲水性β-桶状跨膜蛋白。牦牛ZP3基因编码区的成功克隆,为进一步研究该基因的结构与功能及其在受精过程中的作用提供了基础。     

牦牛和黄牛的睾丸形态及其血管构筑特征

本实验旨在研究牦牛（Bos grunniens）和黄牛（B. taurus）的睾丸形态及其血管构筑和动脉管径特征,为牦牛睾丸在高原环境的生理适应性提供依据。从14头屠宰后的成年牦牛体内采集睾丸28枚,从9头成年黄牛体内采集睾丸18枚,测定其形态指标,利用血管铸型技术制作动静脉构筑标本,研究睾丸血管解剖学及主要动脉管径的特征。结果显示,牦牛和黄牛的睾丸、附睾的形态特征及动脉管径有差异,牦牛大部分睾丸动脉及其分支的管径与睾丸重的相对值极显著地高于黄牛（P < 0.01）,牦牛睾丸的主要动脉构筑特征与黄牛的相同,但其大的集合静脉数量较少,小静脉呈“编织袋”状紧密排布。研究认为,牦牛睾丸的血管解剖特征可为睾丸提供更为充足的血液,其相对发达的动脉血管和睾丸静脉“编织袋”状分布特点有利于睾丸的温度调节及精子成熟,可能是牦牛生殖器官适应高海拔环境的生理特征之一。     

低温驯化对布氏田鼠小肠黏膜结构和黏膜免疫相关细胞的影响

环境温度的变化影响野生啮齿动物的消化道形态与功能。小肠是吸收营养成分的主要部位,其结构和功能具有可塑性。为了解小肠黏膜的结构和功能对环境温度变化的响应机制,以布氏田鼠为研究对象,比较了低温组和常温组动物小肠黏膜的组织结构和小肠黏膜免疫相关细胞的数目。结果显示：（1）低温组布氏田鼠的十二指肠、空肠和回肠的绒毛长度及绒毛长度与隐窝深度的比值均高于对照组;（2）低温驯化使布氏田鼠小肠上皮内淋巴细胞的数量增加;（3）低温驯化使布氏田鼠十二指肠、空肠和回肠的杯状细胞数量均显著增加。结果表明,在低温环境下布氏田鼠的小肠黏膜结构和免疫细胞的数量发生了可塑性变化,这可能与低温环境下的高能量需求和免疫功能的变化有关。     

不同的中和剂对L(+)-乳酸发酵的影响

分别利用CaCO3、6 mol/L氨水和6 mol/L NaOH溶液调控乳酸发酵过程的pH,得到的乳酸浓度为169.1g/L、187.9g/L和170.1g/L(以发酵液的初始体积计算),分别是无pH调控发酵过程的4.9倍、5.4倍和4.9倍;得到的OD620分别为19.3、21.6和16.4,分别是无pH调控(OD620为8.5)的2.3倍、2.5倍和1.9倍.相对于氨水和NaOH来说,CaCO3粉末是一种缓慢型的酸中和剂,pH调节能力有限,只能将pH维持在4.9～5.2.但CaCO3可以将乳酸以生成乳酸钙的形式沉淀下来,给下游乳酸的分离提取带来一定的方便.因此对于传统的分批式发酵,CaCO3仍不失为一种较好的选择.氨水和NaOH溶液都可以很好地将发酵液的pH调控在6.0,其中氨水是一种最理想的酸中和剂,既有利于乳酸的生物合成又能促进乳酸菌的生长.     

GLP-2对小鼠小肠黏膜的作用及不同脏器分布的影响

目的：观察胰高血糖素样肽-2对小鼠小肠黏膜的作用效果及其受体在不同脏器的分布和表达。方法：选用5-6周龄的雄性健康BALB/C小鼠,体重17～20g。随机分为3组。对照组8只;脑内注射GLP-2组9只,每天按25μg/kg脑内注射GLP-2液2次,连续3天;腹腔注射GLP-2组9只,每天按250μg/kg腹腔注射GLP-2液2次,连续3天。3天后,处死小鼠,做组织切片进行HE染色,观察小鼠小肠黏膜的组织学变化,用免疫组化方法检测GLP-2R在不同脏器的表达和分布。结果：小鼠经腹腔注射GLP-2后,小肠不同肠段黏膜的绒毛高度较对照组明显增加（P〈0.05）,脑内注射组的肠黏膜无明显形态学变化;GLP-2R在小鼠胃、空肠与结肠部位均有表达,而食管与肝显阴性;腹腔注射组的GLP-2R表达较对照组显著下调（P〈0.05）。结论：GLP-2能刺激小肠黏膜上皮增厚,增加小肠不同肠段黏膜的绒毛高度;小鼠的胃、空肠与结肠经GLP-2（腹腔注射）处理后其受体表达下调。     

痘苗病毒E3L蛋白的结构与功能

E3L蛋白是痘苗病毒基因组所编码的一种非常重要的毒力蛋白。E3L结构高度保守,其N端为Z-DNA结合域(Zα),C端为双链RNA结合域(dsRBM),两者均为其致病性所必需。E3L具有抵御宿主细胞干扰素及其诱导蛋白的抗病毒作用,以及抑制细胞凋亡、抵抗RNA干扰等多方面的重要功能,这些策略有助于逃避宿主的抗病毒免疫。     

Effect of Weaning on Small Intestinal Structure and Function in the Piglet

Fifty-four piglets were selected from 12 litters weaned at 17 (Treatment 1), 21 (Treatment 2), 28 (Treatment 3) and 35 (Treatment 4) days old, respectively, to determine the effect of weaning age on small intestinal villus morphology, immunology and histochemistry. From proximal duodenum, proximal jejunum, distal jejunum and middle ileum, intestinal samples with three replicates (piglets) in each treatment were taken at 18, 22, 28 and 36; 22, 28, 36 and 43; 28, 36, 43, and 50; and 18, 22, 28, 36, 43 and 50d of age in Treatment 1, 2, 3 and 4, respectively. This was equivalent to 12h, 3d, 1 week, 2 week postweaning in Treatment 1; 12h, 1 week, 2 week, 3 week postweaning in Treatment 2 and 3, and all the same age in Treatment 4 as in Treatment 1, 2, 3, respectively. The results showed that villous height of duodenum and proximal jejunum decreased significantly in Treatment 1 and 3. Crypt depth in the duodenum, proximal jejunum and ileum also decreased significantly in Treatment 1. Date had significant effect on villous height of the duodenum, distal jejunum and ileum with the shortest on day 29 and crypt depth of all positions increased with piglet age except the crypt depth in proximal jejunum decreased on day 50. Weaning age and day of age had significant effects on intraepithelial lymphocyte (IEL) number and goblet cell (GC) number at all positions of small intestinal mucosa in piglets. The number of IEL at all segments of small intestinal mucosa in Treatment 3 increased significantly compared to those in other treatments, but IEL number at all locations of small intestinal mucosa in Treatment 2 decreased significantly compared to those in other treatments. The number of GC in small intestinal mucosa increased significantly in early-weaned (< day 21) piglets. It appears that providing fluid milk replacer for a few days postweaning could dramatically reduce the negative impact of weaning on villous morphology and digestive and absorptive function, especially in pigs weaned prior to 3 week of age. Finally, as weaning age was reduced, GC had a greater role in intestinal duct protection.     

Lactobacillus acidophilus Alleviates Platelet-Activating Factor-Induced Inflammatory Responses in Human Intestinal Epithelial Cells

Probiotics have been used as alternative prevention and therapy modalities in intestinal inflammatory disorders including inflammatory bowel diseases (IBD) and necrotizing enterocolitis (NEC). Pathophysiology of IBD and NEC includes the production of diverse lipid mediators, including platelet-activating factor (PAF) that mediate inflammatory responses in the disease. PAF is known to activate NF-κB, however, the mechanisms of PAF-induced inflammation are not fully defined. We have recently described a novel PAF-triggered pathway of NF-κB activation and IL-8 production in intestinal epithelial cells (IECs), requiring the pivotal role of the adaptor protein Bcl10 and its interactions with CARMA3 and MALT1. The current studies examined the potential role of the probiotic Lactobacillus acidophilus in reversing the PAF-induced, Bcl10-dependent NF-κB activation and IL-8 production in IECs. PAF treatment (5 µM×24 h) of NCM460 and Caco-2 cells significantly increased nuclear p65 NF-κB levels and IL-8 secretion (2-3-fold, P<0.05), compared to control, which were blocked by pretreatment of the cells for 6 h with L. acidophilus (LA) or its culture supernatant (CS), followed by continued treatments with PAF for 24 h. LA-CS also attenuated PAF-induced increase in Bcl10 mRNA and protein levels and Bcl10 promoter activity. LA-CS did not alter PAF-induced interaction of Bcl10 with CARMA3, but attenuated Bcl10 interaction with MALT1 and also PAF-induced ubiquitination of IKKγ. Efficacy of bacteria-free CS of LA in counteracting PAF-induced inflammatory cascade suggests that soluble factor(s) in the CS of LA mediate these effects. These results define a novel mechanism by which probiotics counteract PAF-induced inflammation in IECs.     

Fermentation of coconut water by probiotic strains Lactobacillus acidophilus L10 and Lactobacillus casei L26

Coconut water is becoming an increasingly popular beverage and sports drink in tropical countries due to its high mineral content. Probiotic fermentation of coconut water would provide consumers with a novel probiotic beverage which can provide both hydration and probiotic benefits. The aim of this study was to assess the growth, survival and fermentation performance of two probiotic bacteria in coconut water. Lactobacillus acidophilus L10 and L. casei L26 grew well in coconut water and showed similar growth patterns. The viable cell count of the two probiotic cultures reached approximately 10⁸ CFU/ml after 2 days fermentation at 37 °C and maintained approximately10⁷–10⁸ CFU/ml after 26 days at 4 °C. Changes in total soluble solids (^oBrix), pH, sugars, organic acids and minerals were similar between the two probiotic cultures, except for fructose, glucose, copper, phosphorus and lactic, acetic and malic acids. There were significant variations between the two cultures in their ability to produce and consume these compounds. L. acidophilus produced higher amounts of 2-heptanone, 2-nonanone, benzaldehyde, 2-heptanol, 2-nonanol, δ-octalactone and δ-dodecalactone, whereas L. casei produced higher amounts of acetic acid, diacetyl, acetoin, δ-decalactone, 3-methyl-3-buten-1-ol, linalool, 1-octanol, p-tolualdehyde and ethyl 2-hydroxypropanoate. There was no substantial change in mineral content. These results suggest the feasibility of fermenting coconut water into a probiotic beverage, especially for sports nutrition, with the dual benefits of electrolytes and probiotics.     

Effects of Probiotic Lactobacillus acidophilus Strain INMIA 9602 Er 317/402 and Putative Probiotic Lactobacilli on DNA Damages in the Small Intestine of Wistar Rats In Vivo

Probiotics and Antimicrobial Proteins - Double-strand breaks in the DNA of the small intestine in male Wistar rats were studied using a neutral comet assay after 7&nbsp;days of feeding with a...     

Viable Intestinal Passage of a Canine Jejunal Commensal Strain Lactobacillus acidophilus LAB20 in Dogs

The strain Lactobacillus acidophilus LAB20 with immunomodulatory properties was previously found dominant in the jejunal chyme of four dogs, and the novel surface layer protein of LAB20 suggested its competitive colonization in canine gut. To evaluate the persistence and survival of LAB20 in healthy dogs, LAB20 was fed to five healthy pet dogs for 3 days, at a dosage of 10⁸ CFU daily as fermented milk supplement. The fecal samples, from 1 day prior to feeding, three continuous feeding days, and on day 5, 7, 14, and 21, were collected for strain-specific detection of LAB20 using real-time PCR. We found that LAB20 count was significantly increased in dog fecal samples at the second feeding day, but rapidly decreased after feeding ceased. The fecal samples from prior to feeding, during feeding, and post-cessation days were plated onto mLBS7 agar, from where LAB20 was recovered and distinguishable from other fecal lactobacilli based on its colony morphotype. Using strain-specific PCR detection, the colonies were further verified as LAB20 indicating that LAB20 can survive through the passage of the canine intestine. This study suggested that canine-derived strain LAB20 maintained at high numbers during feeding, viably transited through the dog gut, and could be identified based on its colony morphotype.