秦岭滑蜥断尾再生伤口愈合阶段细胞因子的时空分布特征

细胞因子参与免疫应答、介导炎症反应,进而在损伤组织的再生修复过程中发挥调控作用。通过切割法建立秦岭滑蜥（Scincella tsinlingensis）断尾再生模型,针对伤口愈合阶段,采用免疫组织化学方法检测促炎症因子白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-1β(IL-1β)、白细胞介素-1受体1(IL-1R1)、肿瘤坏死因子-α(TNF-α)和抗炎细胞因子白细胞介素-10(IL-10)的组织细胞定位及表达变化。结果显示：6种细胞因子在断尾脊髓和椎骨中的骨系细胞中均呈阳性。断尾创面组织及其附近鳞片下方真皮层中IL-6阳性细胞数量先增加后减少,断尾后3 d达峰值194.3±24.9。断尾后3 d,创面IL-8阳性细胞数目为107.7±12.7,超过真皮层的43.7±9.1。IL-1β和IL-1R1阳性细胞数量先增后减,伤口部位的阳性细胞明显多于真皮层（P<0.05）。TNF-α阳性细胞由0.5 d的126.3±35.0增至3 d的190.3±12.1。IL-10阳性细胞断尾后总体呈现先增后减的趋势,断尾后0.5 d伤口部位阳性细胞达201±17.8,显著...     

Ghrelin、KGF、TGF-β(β2、β3)在大鵟胃肠道中的表达

应用免疫组织化学方法和体视学半定量方法,检测了ghrelin、KGF、TGF-β(TGF-β_2,TGF-β_3)在大鵟(Buteo hemilasius)胃肠道中的表达,利用IPP专业图像分析软件对其表达强度进行了定量分析.结果表明,ghrelin免疫反应阳性物质分布在十二指肠、空肠、回肠、盲肠和直肠的黏膜层,主要分布于黏膜上皮、肠腺上皮和固有层.从十二指肠到盲肠阳性细胞的分布密度逐渐减小,直肠阳性细胞的分布密度高于盲肠.胃肠道呈KGF免疫反应阳性,胃内阳性物质分布于腺胃浅腺和深腺、肌胃黏膜、肌胃单管腺的上皮细胞;肠内阳性物质分布于固有层的血管、淋巴和平滑肌纤维.胃黏膜、腺胃深腺、肌胃单管腺、肠道黏膜、肠腺呈TGF-β_2及TGF-β_3免疫反应阳性,阳性物质分布于黏膜、肠腺上皮细胞胞质中.图像分析显示,KGF的阳性表达水平呈波浪形变化,在空肠和直肠处达到峰值;TGF-β2的阳性表达水平呈波浪形变化,分别在肌胃和空肠处有峰值;TGF-β_3从腺胃到空肠阳性表达水平逐渐增强,之后阳性表达水平又逐渐下降,到盲肠阳性表达水平回升.ghrelin、KGF、TGF-β_2和TGF-β_3的阳性表达强弱可能与胃肠道的消化能力有关,它们的协同表达调控鸟类胃肠道的生长和发育.     

肺炎链球菌与宿主炎症小体的相互作用机制

肺炎链球菌(Streptococcus pneumoniae)是一种定植于上呼吸道的革兰阳性胞外菌,是导致侵袭性肺炎的主要原因,所致疾病具有较高的发病率和死亡率。炎症小体(inflammasome)是胞浆内重要的蛋白复合体,在先天免疫应答过程中起着重要作用。大量研究表明,肺炎链球菌感染可诱导宿主炎症小体的激活、半胱天冬酶1的活化和促炎性细胞因子的分泌。在长期选择压力的作用下,肺炎链球菌的部分突变菌株可以逃避炎症小体的识别。本文就肺炎链球菌感染过程中炎症小体的激活、炎症小体在抗肺炎链球菌过程中的作用以及肺炎链球菌逃避宿主炎症小体识别的机制三方面对肺炎链球菌与炎症小体之间相互作用的研究进展进行综述。     

自然感染途径建立肺炎链球菌感染性肺炎小鼠模型

目的探索自然感染途径制备肺炎链球菌感染性肺炎小鼠模型的方法,为肺炎链球菌性肺炎的相关研究提供实验基础。方法肺炎链球菌标准菌株(CMCC 31203)经血平板培养18 h后配成2麦氏浊度。小鼠浅麻醉状态下,破损鼻黏膜,滴注一定量肺炎链球菌菌液。于3、7、14和21 d分别处死小鼠,取肺脏进行组织病理学观察,确定肺炎小鼠模型制备的最佳方式。结果鼻黏膜破损组小鼠3 d时肺泡间隔内毛细血管扩张,肺泡腔内以红细胞和纤维素渗出为主;7 d时病变以纤维素和中性粒细胞浸润为主;14 d时肺泡腔内渗出减少,炎症开始减轻;21 d时肺脏外观趋于正常,肺泡腔内渗出物质基本溶解吸收。结论以3×107CFU的肺炎链球菌菌液通过破损的鼻黏膜感染7 d时可以制备出比较典型、稳定的肺炎小鼠模型。     

miR-124-3p靶向TLR4抑制流感病毒性肺炎小鼠的炎症反应

目的:探讨miR-124-3p靶向Toll样受体4(TLR4)对流感病毒性肺炎小鼠炎症反应的影响。方法:建立流感病毒亚洲甲型鼠肺适应株(FM1)感染所致病毒性肺炎小鼠模型,采用RT-qPCR测定肺组织miRNA-124-3p表达水平。病毒性肺炎小鼠尾部注射miR-124-3p agomir或agomir NC后,第4 d摘眼球取血并取出肺组织,ELISA检测血浆中白细胞介素(IL)-1β、IL-6、肿瘤坏死因子α(TNF-α)水平,HE染色观察肺组织病理变化,Western印迹测定TLR4和核转录因子κB(NF-κB)p65蛋白水平。结果:与正常肺组织比较,病毒性肺炎小鼠肺组织miR-124-3p表达量降低(P0.001);miR-124-3p agomir注射组小鼠血浆中IL-1β、IL-6、TNF-α明显低于注射agomir NC组和模型组(P0.001),而高于正常组;HE染色结果表明miR-124-3p agomir注射组小鼠的肺组织炎细胞渗出和浸润较agomir NC组和模型组减轻;与agomir NC组比较,miR-124-3p agomir注射组的TLR4、NF-κB p65表达减少(P0.01)。结论:miR-124-3p抑制TLR4/NF-κB信号通路减轻流感病毒性肺炎小鼠的炎症反应。     

炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中的表达及临床意义

目的:探讨炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中的表达水平及其临床意义。方法:收集2014年6月至2016年6月我院儿科收治的79例大叶性肺炎患儿为病例组,按照临床肺部感染评分(CPIS)分为重度组(n=46)和轻度组(n=33),于同期随机选取30例健康儿童为对照组,采用双抗体夹心酶联免疫吸附法(ELISA)检测各组支气管肺泡灌洗液及血清炎症因子如白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-17(IL-17)、肿瘤坏死因子(TNF-α)以及高迁移率族蛋白B1(HMGB1)表达水平。结果:重度组、轻度组血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于对照组,重度组血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于轻度组,差异有统计学意义(P0.05)。病例组患儿支气管肺泡灌洗液IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于血清,差异有统计学意义(P0.05)。重度组、轻度组支气管肺泡灌洗液IL-6、IL-8、IL-17、TNF-α及HMGB1水平高于血清,差异有统计学意义(P0.05),重度组支气管肺泡灌洗液、血清IL-6、IL-8、IL-17、TNF-α及HMGB1水平分别高于轻度组,差异有统计学意义(P0.05)。经Spearman积矩相关分析,支气管肺泡灌洗液与血清中IL-6、IL-8、IL-17、TNF-α及HMGB1水平分别呈正相关关系(r=0.711、0.695、0.752、0.793、0.728,P0.05)。结论:炎症因子在大叶性肺炎患儿支气管肺泡灌洗液及血清中呈高表达,并与大叶性肺炎病情严重程度密切相关,联合检测有助于早期评估病情。     

肺炎衣原体感染与急性心肌梗死关系的试验研究及初探

目的检测血清肺炎衣原体(TWAR)和白细胞介素-6 (IL-6)对急性心肌梗死(AMI)的诊断价值及血清脂质谱的变化,以期了解我国AMI患者TWAR的感染状况以及相关因子变化的临床意义.方法采用间接显微免疫荧光法检测53例急性心肌梗死病人和50例健康体验者血清肺炎衣原体IgG和IgM滴度,用ELISA法分别检测其血清白细胞介素-6(IL-6)的含量,同时检测两组血清脂质谱的含量.结果急性心肌梗死组肺炎衣原体既往感染率(73.6%)明显高于对照组(14%)(P ＜0.01),两组肺炎衣原体急性感染率差异无显著性(P＞0.05);两组血清中IL-6 含量超过正常上限值的阳性率差异有显著性,即对照组明显低于急性心肌梗死组(P＜0.0 1).发现急性心肌梗死组肺炎衣原体感染阳性者,其危险的血清脂质谱较对照组有明显的增加.结论肺炎衣原体感染与冠心病和急性心肌梗死的发生、疾病发展过程有密切关系 .     

经肺炎衣原体感染的SD大鼠的病理学特征变化分析研究

摘要 目的：探讨与分析大鼠肺炎衣原体感染后的病理学特征变化。方法：研究时间为2019年5月到2020年2月。将30只斯泼累格?多雷(Sprague Dawley,SD)大鼠随机分为2组-实验组与对照组,每组各15只大鼠。实验组大鼠从鼻腔吸入40 μL 含1×103感染颗粒的肺炎衣原体,对照组吸入等剂量的无菌磷酸液缓冲液。观察与检测大鼠一般行为、血液学指标与病理学变化情况。结果：实验组肺实变面积达25 %~50 %,细支气管和小血管周围出现小灶性淋巴细胞及单个核细胞聚集,肺泡腔有大量炎性渗出,肺泡壁伴随有充血,支气管周围见大量嗜中性粒细胞浸润。小鼠一般行为表现为活力下降,毛发皱乱,进食和饮水减少,进食明显减少。接种后3 d,实验组的白细胞总数、中性粒细胞比例高于对照组,淋巴细胞比例低于对照组(P<0.05);实验组的血清白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)浓度都显著高于对照组(P<0.05);实验组的肺炎衣原体IgG抗体相对表达水平显著高于对照组(P<0.05);实验组的血清血管内皮生长因子(Vascular endothelial growth factor,VEGF)、D-二聚体(D-dimer,D-D)表达水平都显著高于对照组(P<0.05)。结论：大鼠肺炎衣原体感染后伴随有肺组织病理损伤,也可诱发VEFG与D-D的表达,促进肺组织炎症细胞浸润,可导致大鼠白细胞总数、中性粒细胞比例增加,促进炎症因子的释放。     

难治性肺炎支原体肺炎患儿血清炎性细胞因子变化的临床意义及其危险因素分析

目的:探讨难治性肺炎支原体肺炎(RMPP)患儿血清炎性细胞因子变化的临床意义及其危险因素。方法:将我院从2016年1月～2020年1月收治的72例RMPP患儿纳入研究,记作RMPP组。另取我院同期收治的普通肺炎支原体肺炎(MPP)患儿70例作为对照组。检测所有患儿的血清炎性细胞因子水平,并进行对比。此外,对比两组患儿各项基线资料,并以多因素Logistic回归分析RMPP发生的危险因素。另外,比较两组患儿的影像学特征。结果:RMPP组患儿血清白细胞介素-6(IL-6)以及白细胞介素-10(IL-10)水平均高于对照组(P0.05);而两组肿瘤坏死因子-α(TNF-α)水平对比差异不明显(P0.05)。RMPP组年龄、发热持续时间、发热峰值、住院时间以及发热伴随症状人数占比均高于对照组(P0.05)。经多因素Logistic回归分析发现:发热持续时间、发热峰值、住院时间以及发热伴随症状均是RMPP患儿发病的危险因素(OR=2.773、2.344、1.058、2.515,均P0.05)。RMPP组大叶性肺炎、胸腔积液人数占比均高于对照组,而支气管肺炎人数占比低于对照组(P0.05)。结论:RMPP患儿血清IL-6以及IL-10水平均存在明显高表达,且发热持续时间、发热峰值、住院时间以及发热伴随症状均是RMPP发病的危险因素,临床工作中需要予以重视。     

重症肺炎患者干扰素-r、IL-6 和TNF-alpha含量变化研究

目的：探究重症肺炎患者干扰素-r、白细胞介素-6 和肿瘤坏死因子-alpha含量变化及其临床意义。方法：选取我院收治并确诊 为肺炎的患者103 例,根据病情不同,将其分为重症肺炎组52 例及普通肺炎组51 例,同时选取同期参加健康体检人群50 例,为 对照组。比较三组成员不同时间段干扰素-r、白细胞介素-6 和肿瘤坏死因子-alpha含量,重症肺炎组患者存活与死亡组上述因子含 量情况。结果：入院6 h 后,与对照组比较,重症肺炎及普通肺炎组干扰素-r、白细胞介素-6 及肿瘤坏子因子-琢含量较高P＜ 0.05,与普通肺炎组比较,重症肺炎组重症肺炎组干扰素-r、白细胞介素-6 及肿瘤坏子因子-alpha含量较高P＜0.05;入院24 h后,与 对照组比较,重症肺炎组干扰素-r、白细胞介素-6 及肿瘤坏子因子-alpha含量较高P＜0.05,普通肺炎组与对照组比较无差异P＞ 0.05;重症肺炎死亡组患者干扰素-r、白细胞介素-6 及肿瘤坏子因子-alpha含量较高P＜0.05。结论：干扰素-r、白细胞介素-6 以及 肿瘤坏死因子-alpha参与患者的免疫调节,可间接提示患者的病程发展,可作为判断病情程度的有利依据。     

Immunohistochemical analysis of ZnT1, 4, 5, 6, and 7 in the mouse gastrointestinal tract.

Expression of five zinc transporters (ZnT1, 4, 5, 6, and 7) of the Slc30 family in the mouse gastrointestinal tract was studied by immunohistochemical analysis. Results demonstrated unique expression patterns, levels, and cellular localization among ZnT proteins in the mouse gastrointestinal tract with some overlapping. ZnT1 was abundantly expressed in the epithelium of the esophagus, duodenum of the small intestine, and cecum of the large intestine. ZnT4 was predominantly detected in the large intestine. ZnT5 was mainly expressed in the parietal cell of the stomach and in the absorptive epithelium of the duodenum and jejunum. ZnT6 was predominantly detected in the chief cell of the stomach, columnar epithelial cells of the jejunum, cecum, colon, and rectum. Lastly, ZnT7 was observed in all epithelia of the mouse gastrointestinal tract with the highest expression in the small intestine. Expression of ZnT proteins in the absorptive epithelial cell of the gastrointestinal tract suggests that ZnT proteins may play important roles in zinc absorption and endogenous zinc secretion.     

Expression and localization of aquaporins in rat gastrointestinal tract

A family of water-selective channels, aquaporins (AQP), has beendemonstrated in various organs and tissues. However, the localizationand expression of the AQP family members in the gastrointestinal tracthave not been entirely elucidated. This study aimed to demonstrate theexpression and distribution of several types of the AQP family and tospeculate on their role in water transport in the rat gastrointestinal tract. By RNase protection assay, expression of AQP1-5 and AQP8 was examined in various portions through the gastrointestinal tract.AQP1 and AQP3 mRNAs were diffusely expressed from esophagus to colon,and their expression was relatively intense in the small intestine andcolon. In contrast, AQP4 mRNA was selectively expressed in the stomachand small intestine and AQP8 mRNA in the jejunum and colon.Immunohistochemistry and in situ hybridization demonstrated cellularlocalization of these AQP in these portions. AQP1 was localized onendothelial cells of lymphatic vessels in the submucosa and laminapropria throughout the gastrointestinal tract. AQP3 was detected on thecircumferential plasma membranes of stratified squamous epithelialcells in the esophagus and basolateral membranes of cardiac glandepithelia in the lower stomach and of surface columnar epithelia in thecolon. However, AQP3 was not apparently detected in the smallintestine. AQP4 was present on the basolateral membrane of the parietalcells in the lower stomach and selectively in the basolateral membranesof deep intestinal gland cells in the small intestine. AQP8 mRNAexpression was demonstrated in the absorptive columnar epithelial cellsof the jejunum and colon by in situ hybridization. These findings mayindicate that water crosses the epithelial layer through these waterchannels, suggesting a possible role of the transcellular route forwater intake or outlet in the gastrointestinal tract.     

Expression,localization and possible functions of aquaporins 3 and 8 in rat digestive system

Although aquaporins (AQPs) play important roles in transcellular water movement, their precise quantification and localization remains controversial. We investigated expression levels and localizations of AQP3 and AQP8 and their possible functions in the rat digestive system using real-time polymerase chain reactions, western blot analysis and immunohistochemistry. We investigated the expression levels and localizations of AQP3 and AQP8 in esophagus, forestomach, glandular stomach, duodenum, jejunum, ileum, proximal and distal colon, and liver. AQP3 was expressed in the basolateral membranes of stratified epithelia (esophagus and forestomach) and simple columnar epithelia (glandular stomach, ileum, and proximal and distal colon). Expression was particularly abundant in the esophagus, and proximal and distal colon. AQP8 was found in the subapical compartment of columnar epithelial cells of the jejunum, ileum, proximal colon and liver; the most intense staining occurred in the jejunum. Our results suggest that AQP3 and AQP8 play significant roles in intestinal function and/or fluid homeostasis and may be an important subject for future investigation of disorders that involve disruption of intestinal fluid homeostasis, such as inflammatory bowel disease and irritable bowel syndrome.     

蓝狐消化道内分泌细胞的免疫组化研究

蓝狐又名北极狐(Alopex lagopus),属于食肉目(Carnivora),犬科(Canidae),北极狐属(Alopex).     

Gastrointestinal infection with Mexican TcI Trypanosoma cruzi strains: different degrees of colonization and diverse immune responses

Mexican Ninoa and Queretaro (Qro) TcI strains of Trypanosoma cruzi have shown different degrees of virulence, and the two strains produce heterogeneous immune responses in the hearts of infected mice. This work shows that the same strains can invade the intestine by an intraperitoneal route and establish an infection, mainly in the colon. The three segments of the small intestine (duodenum, jejunum and ileum) were infected to a lesser degree than the colon. Despite the fact that parasites were predominantly found in the colon, an obvious inflammatory reaction was observed in the submucosal layer along the entire intestinal tract, with the virulent Qro strain causing significantly more areas of higher immune infiltration. A clear recruitment of CD4+ and CD8+ T lymphocytes to the mesenteric ganglia was observed during infection with the virulent strain. Macrophages were also differentially distributed in the gastrointestinal tract. These later cells infiltrated fewer amastigote nests in the mice infected with the Qro strain than in the mice infected with the Ninoa strain. When IFN-γ, TNF-α, and IL-4 levels were measured, an increase in these cytokines was observed compared with the uninfected mice. The role of these inflammatory reactions in the pathogenesis of Chagas enteropathy is also discussed in this paper.     

Reaction and response of newborn guinea pigs to experimental Salmonella typhi infection

Newborn guinea pigs, orally infected with Salmonella typhi were examined at various intervals of time in order to determine bacterial distribution in tissues and to establish possible correlation with the clinical aspects manifested. Histopathological examination evidenced typical lesions in jejunum, ileum, caecum and especially in regional lymphatic tissues. Spleen, liver and mesenteric lymph nodes presented granulomatous lesions similar to those observed in in human typhoid fever. After oral administration, the animals reacted with anorexia, febrile reactions, bacteremia, diarrhoea, positive stool cultures, dehydration, lethargy and antibodies too were produced. Our results indicate that typhoid infection may be induced in newborn guinea pigs; the model may be used for an assessment of attenuated live typhoid vaccine control.     

Kallikrein-gene expression in the rat gastrointestinal tract.

The serine proteinase glandular kallikrein has been demonstrated in the gastrointestinal tract, although there is some doubt as to whether it is synthesized there or derives from exocrine-gland secretions. Using a rat pancreatic kallikrein cRNA probe we have demonstrated kallikrein-like gene expression in the corpus, duodenum, jejunum, ileum, caecum and colon, and compared the pattern of expression with that of the gastrointestinal peptides somatostatin, gastrin and glucagon. In addition, using a panel of oligonucleotide probes specific for various members of the rat kallikrein-gene family, we have shown that the kallikrein-like gene expressed appears to be expressed as true kallikrein.     

Constitutive production of lymphocyte activating factors by normal tissues in the adult rat

Lymphocyte activating factors (LAFs), e.g., interleukin-1 (IL-1) and IL-1-like factors, have previously been demonstrated outside the immune system in the skin, thymus epithelium, and the human and rat testis. We have studied the presence of LAFs in normal tissues of the adult rat, utilizing a highly IL-1 sensitive murine thymocyte proliferation assay. We have demonstrated high amounts of LAF activity in the tongue, esophagus, proventricular part of the stomach, and the liver. Some activity was also demonstrated in the duodenum, placenta, spleen, Peyer's patches, glandular stomach, and jejunum, but no bioactivity was present in other gastrointestinal, endocrine, lymphoid, or haematopoeitic tissues. We were also unable to detect any LAF activity in the reproductive organs (except for the testis), urinary tract, skeletal and muscular tissues, brain, eyes, salivary glands, or lung. In the esophagus the activity was mainly localized to the mucosa. The LAF activity in the skin was partly inhibited by treatment with a mixture of antibodies against human IL-1 alpha and IL-1 beta. Dose response curves and gel filtration on a Sephacryl S-200 column suggested the presence of a high molecular weight (90,000-100,000 Da) LAF inhibitory factor in the liver. In all positive tissues, the demonstrated LAFs had a molecular weight of 15,000-25,000 Da, as determined by Sephacryl S-200 gel filtration. Of the positive tissues, the skin, tongue, esophagus, and the proventricular part of the stomach all contain stratified squamous epithelium. It is tempting to suggest that the detected LAFs have a similar function in these barrier tissues, e.g., to serve as host defence factors, or, alternatively or additionally, as tissue growth factors.