大鳞副泥鳅肠道黏液细胞及消化酶活性分布特征研究

采用阿利新兰-碘酸雪夫氏反应(AB-PAS)染色法及酶学方法研究了大鳞副泥鳅成熟个体肠道各段黏液细胞分布及消化酶活性。结果表明, 大鳞副泥鳅肠道黏液细胞分为Ⅰ、Ⅱ、Ⅲ和Ⅳ 4种类型。前肠至后肠, 黏液细胞数量逐渐减少。前肠主要分布Ⅲ和Ⅳ混合型黏液细胞, 后肠则以Ⅱ和Ⅳ型酸性黏液细胞为主。肠道胰蛋白酶活性显著高于淀粉酶和脂肪酶。且后肠消化酶活性显著低于前肠和中肠。根据黏液细胞及消化酶活性分布特点, 表明大鳞副泥鳅属于杂食性鱼类, 前肠为其主要的消化吸收场所, 后肠中性黏液细胞的数量较少以及消化酶活性较低, 表明其对食物的消化吸收功能较弱, 与其为辅助呼吸功能的特点相关。     

四指马鲅稚鱼、幼鱼和成鱼消化道黏液细胞组织化学研究

采用常规石蜡组织切片及阿利新兰–高碘酸雪夫–试剂(AB-PAS,AB,p H2.5)染色方法对四指马(Eleutheronema tetradactylum)稚鱼、幼鱼和成鱼消化道中黏液细胞的组织化学特征进行观察。35日龄稚鱼消化道黏液细胞数量少,种类单一,食道含数量较多的II型黏液细胞,偶见I型黏液细胞;幽门盲囊未检测出黏液细胞的分布;前肠以I型黏液细胞为主,有少量III型黏液细胞、中肠和后肠均只有II型黏液细胞。65日龄幼鱼消化道各段黏液细胞数量明显增多,种类丰富,食道、前肠和中肠均含I、II、III、IV型4种类型的黏液细胞,其余均含至少两种类型的黏液细胞,食道黏液细胞密度最大,其次是后肠。成鱼食道、幽门盲囊均含II型和IV型黏液细胞,但后者两型黏液细胞数量都比较少;胃部含有大量I型黏液细胞,胃腺区域则以IV型黏液细胞为主;肠道只含III、IV型黏液细胞,黏液细胞密度从前到后呈递增规律。稚鱼、幼鱼、成鱼消化道黏液细胞的分布特点体现消化道结构功能逐渐发育完善的规律。     

黄颡鱼仔稚鱼胃肠发育的显微和超微结构研究

利用光学显微技术和透射电镜技术,观察和研究了出膜后1-35日龄黄颡鱼(Pelteobagrus fulvidraco)仔稚鱼的胃肠发育.水温为23-25℃时,2日龄仔稚鱼的消化道分化出口咽腔、食道、胃、肠;3日龄肠道分化为前肠、中肠、后肠.3日龄黄颡鱼开口摄食时其胃贲门部黏膜层下出现胃腺,为已有鱼类研究报道中胃腺最早出现的日龄.超微结构显示3日龄胃腺细胞中可见胃蛋白酶原颗粒和丰富的管泡系统,为典型的泌酸胃酶细胞;随日龄增加,胃蛋白酶原颗粒越来越丰富而管泡系统越来越不明显.3日龄时前肠吸收细胞胞质中可见脂肪泡,后肠吸收细胞胞质中可见蛋白质胞饮体.直到25日龄后肠吸收细胞胞质中尚可见蛋白质胞饮体.以七结果表明黄颡鱼在3日龄开口摄食时消化道具备细胞外消化功能,但此功能不完善,期间继续通过胞饮作用等细胞内消化来弥补胞外消化的不足,直到25-30日龄后细胞外消化功能发育完善.采用符合其生理机能发育过程的投喂管理策略可以有效提高大规格苗种培育的成活率.     

鳜早期味蕾发育的组织学特征

采用石蜡切片、H.E染色研究了3 ~ 28日龄鳜（Siniperca chuatsi）味蕾发育的组织学特征,并通过扫描电镜观察28日龄鳜口咽腔组织味蕾类型与数目。结果表明,未开口期（3日龄）,鳜口裂未张开,味蕾尚未分化;开口期（7日龄）,鳜口裂张开明显,味蕾呈椭圆形,突起高度平缓,主要分布在上下颌上皮上,舌、咽、鳃弓上皮上有少量分布;稚鱼期（14日龄）,味蕾呈圆锥形,突起高度上升,舌和咽上味蕾数目增加;21日龄,味蕾呈近梯形,突起高度不变,下颌、舌、咽上味蕾数增加,鳃弓上味蕾数目显著增加;28日龄,味蕾发育完全,口咽腔味蕾数继续增加。扫描电镜观察表明,鳜味蕾主要有3种类型：Ⅰ型味蕾近球形,含有大量微绒毛,突起高于上皮,味孔向外突起;Ⅱ型味蕾含有少量微绒毛,突起略高于黏膜上皮,味孔向内凹陷;Ⅲ型味蕾微绒毛含量最少,突起几乎与黏膜上皮共面,味孔平坦或凹陷。上下颌、咽、鳃中以Ⅰ型味蕾数量最多,Ⅱ型味蕾最少,舌上主要分布Ⅰ型味蕾,无Ⅲ型味蕾。结果表明,鳜早期味蕾结构发育与其摄食关联,推测其主要通过Ⅰ型味蕾和Ⅱ型味蕾对食物的机械性和化学成分进行识别。     

两种海马鳃组织和消化道的黏液细胞类型及分布

运用阿新兰(AB,pH 2.6)和过碘酸雪夫氏(PAS)反应染色方法,对三斑海马(Hippocampus trimaculatus)和日本海马(H.japonicus)鳃组织与消化道中的黏液细胞类型及分布进行了研究。染色结果显示:两种海马的鳃组织和消化道中均含有黏液细胞,日本海马的鳃组织中含有Ⅰ型和Ⅳ型黏液细胞,三斑海马的鳃组织中含有Ⅰ型、Ⅲ型和Ⅳ型黏液细胞。两种海马消化道各部位的黏液细胞类型和数量有明显差异:日本海马的食道中Ⅰ型细胞最多,而三斑海马的食道中Ⅳ型细胞最多;日本海马的前肠中只含有Ⅰ型细胞,而三斑海马的前肠中含有Ⅰ型、Ⅲ型和Ⅳ型细胞,其中Ⅰ型细胞含量最多;日本海马的中肠中含有Ⅰ型、Ⅲ型和Ⅳ型细胞,其中Ⅲ型细胞含量最多,而三斑海马中肠中只含有Ⅰ型细胞;日本海马与三斑海马的后肠中都分布有Ⅰ型、Ⅱ型、Ⅲ型和Ⅳ型细胞,两者不同的是,日本海马的后肠中Ⅲ型细胞含量最多,三斑海马的后肠中Ⅳ型细胞含量最多。     

银鲳消化道形态与组织学结构特征及其消化酶活性

为了解银鲳(Pampus argenteus)消化道结构特点与其功能及食性的相关性, 采用解剖、石蜡切片、AB-PAS染色及酶活性检测技术对银鲳消化道的形态、组织结构、黏液细胞分布及消化酶活性进行研究。结果显示, 银鲳的消化道由口咽腔(舌)、食道侧囊、食道、胃及肠构成, 胃肠交界处有很多幽门盲囊。食道侧囊呈椭球形, 食道粗短, 胃呈U型, 肠有多个盘曲, 肠指数为2.03。舌上皮内有少量味蕾及较多黏液细胞。食道侧囊、食道、胃及肠均由黏膜层、黏膜下层、肌层及浆膜组成。食道侧囊内皱襞较发达, 被覆复层扁平上皮, 内含较多黏液细胞, 且以Ⅳ型为主, 皱襞顶端及侧面有内含角质刺的次级突起; 黏膜下层及肌层中有固定皱襞的骨质脚根; 侧囊内胃蛋白酶活性较高。食道内皱襞较高, 被覆复层扁平上皮, 内含较多黏液细胞, 且以Ⅳ型为主。胃内皱襞发达, 被覆单层柱状上皮, 未见黏液细胞分布; 胃腺发达, 胃内蛋白酶活性较高。肠道内褶襞多, 高度呈先下降后上升趋势, 黏液细胞密度前、中肠较高, 后肠较低, 且均以Ⅰ型为主; 肠道内胰蛋白酶、脂肪酶、淀粉酶及碱性磷酸酶活性较高。幽门盲囊组织结构与肠相似。银鲳的消化道结构特点、黏液细胞分布及消化酶活性与其功能及偏肉食的杂食性相适应。     

人工养殖稀有鮈鲫消化道组织学观察

采用活体解剖和显微技术对人工养殖的稀有鮈鲫消化道组织结构进行了详细观察,并描述了其形态结构。结果表明:稀有鮈鲫为杂食性无胃鱼,肠道系数0.64±0.06。消化道包括口咽腔、食道、肠和肛门。口咽腔和食道粘膜层为复层扁平上皮,内含较多杯状细胞、粘液细胞和少量味蕾;食道粗短,肌肉层发达。肠由前肠、中肠和后肠三部分组成。肠道由前到后,粘液细胞数量逐渐增多,粘膜皱褶数量逐渐减少,粘膜皱褶高度逐渐降低。     

蜡皮蜥消化系统组织学初步研究

应用石蜡常规切片,HE染色,对蜡皮蜥的消化系统组织结构进行了观察。结果表明:消化道管壁除口咽腔外,其余由粘膜层、粘膜下层、肌层和浆膜层组成;不同部位的消化道管壁厚度各不相同,其中胃壁最厚,幽门胃壁厚达658.0μm,其次为大肠和食道,分别为393.3μm和215.9μm,小肠壁最薄,约为126.8μm;皱襞的发达程度也随消化道的部位不同而有所差异,十二指肠皱襞数量最多,约为27~42个,平均高度为733.2μm,其次分别为回肠、口咽腔和食道,大肠皱襞数量最少约为3~11个,平均高度为484.2μm;食道上皮为复层鳞状上皮细胞,口咽腔、胃、小肠和大肠上皮均为单层柱状上皮;食道和大肠上皮的杯状细胞较多,密度分别为927个/mm2、899个/mm2;肝脏中色素细胞发达并含大量深色颗粒,胰脏泡心细胞较少且胰岛不发达,肝脏小叶和胰脏小叶界限不清晰。这充分说明了蜡皮蜥消化系统的组织结构特征与其食性和生态习性相适应,与鸟类和哺乳动物相比,具有一定的不完善性和低等性。     

三种底栖淡水鱼类皮肤黏液细胞分布与数量比较

采用常规石蜡切片以及AB-PAS染色方法研究了葛氏鲈塘鳢（Perccottus glenii）、黄颡鱼（Pelteobagrus fulvidraco）以及泥鳅（Misgurnus anguillicaudatus）3种底栖淡水鱼类的皮肤黏液细胞类型以及分布,计数10个视野下（视野面积为43.5 μm × 32.6 μm）3种鱼类头部、背部、腹以及尾部皮肤的黏液细胞数量,并用单因素方差分析（ANOVA）比较鱼体4种黏液细胞数量差异。结果表明：（1）3种鱼类的主要黏液细胞不同,葛氏鲈塘鳢鱼体黏液细胞中Ⅲ型细胞居多,较Ⅰ型细胞多61.5%,较Ⅱ型细胞多85.8%,较Ⅳ型细胞多85.7%;在黄颡鱼体表,Ⅰ型黏液细胞分布数量最多,较Ⅱ型细胞多9.9%,较Ⅲ型细胞多15.1%,较Ⅳ型细胞多53.5%;而在泥鳅体表以Ⅱ型细胞数量最为丰富,较Ⅰ型细胞多88.3%,较Ⅲ型细胞多33.1%,较Ⅳ型细胞多83.5%。（2）对于葛氏鲈塘鳢,黏液细胞集中分布在头部,比背部黏液细胞数量多15.4%,比腹部黏液细胞数量多出38.0%,比尾部黏液细胞数量多56.7%;黄颡鱼以背部黏液细胞数量为多,比头部黏液细胞数量多42.5%,比腹部黏液细胞数量多46.6%,比尾部黏液细胞数量多51.4%;泥鳅也在背部具有丰富的黏液细胞,比头部黏液细胞数量多49.9%,比腹部黏液细胞数量50.6%。（3）3种鱼类之间的黏液细胞总数不同,泥鳅体表的平均黏液细胞数量最多,相较于葛氏鲈塘鳢多38.9%,较黄颡鱼多39.1%。研究结果表明,不同鱼类的鱼体表面黏液细胞种类不同,可能与其生活环境和鱼体本身的特性有关。     

不同饵料对稀有鳇鲫仔稚鱼生长、消化道及消化酶的影响

为了筛选稀有逗鲫(Gobiocypris rarus)仔稚鱼期合适的饵料, 将初孵仔鱼随机分为3个组: 活饵组、饲料组、转食组, 每组3个平行, 均从3 dah(day after hatching)开始投喂相应饵料, 在10、15、20、25、30、35 dah时统计生长指标和存活率, 并进行35 dah仔稚鱼消化道组织学及消化酶活性研究。结果显示: (1)35 dah时, 活饵组存活率最高; (2)从15 dah开始, 活饵组表现出最好的生长性能, 并维持这种趋势; (3)活饵组的消化道各部肌层厚度、黏液细胞数目均最高, 提示该组仔稚鱼消化道发育状态较好, 并具有较优的消化吸收能力; (4)转食组胰蛋白酶活性显著高于活饵组和饲料组。研究结果提示: 相对于饲料和转食, 一直投喂活饵更有利于稀有逗鲫仔稚鱼的生长和发育。建议在标准化养殖过程中, 对仔稚鱼期的稀有逗鲫采取活饵投喂的方式。     

Production of arsine and methylarsines in soil and in culture

Arsenate, arsenite, monomethylarsonate, and dimethylarsinate were added to different soils, and evolution of gaseous arsenical products was determined over 3 weeks. Arsine was produced in all three soils from all substrates, whereas methylarsine and dimethylarsine were produced only from methylarsonate and dimethylarsinate, respectively. At least three times more arsine than dimethylarsine was produced in soil incubated with dimethylarsinate. Resting cell suspensions of Pseudomonas and Alcaligenes produced arsine as the sole product when incubated anaerobically in the presence of arsenate or arsenite. In all instances, no trimethylarsine was observed, nor could any evidence be shown for the methylation of any arsenical substrate in soil or in culture. It was concluded that reduction to arsine, not methylation to trimethylarsine, was the primary mechanism for gaseous loss of arsenicals from soil.     

Degradation and conversion of somatostatin in normal and diabetic rats in vivo and in vitro

Plasma somatostatin-like immunoreactivity in the portal and jugular veins of streptozotocin diabetic rats was compared with that in normal control rats. In the diabetic group, somatostatin levels in the portal (p less than 0.05) and jugular (p less than 0.01) veins were both elevated compared with those in the control group. Moreover, the degree of elevation was greater in the jugular vein than in the portal vein. To further investigate the role of the liver in the clearance of somatostatin-28 in vivo, 2 micrograms of somatostatin-28 was administered as a bolus into the external jugular vein of intact and functionally hepatectomized rats. The mean half-time of somatostatin-28 was significantly longer in intact diabetic rats than in controls (p less than 0.05). The functional hepatectomy did not cause a significant difference in the half-time in diabetic rats but made it longer in control rats. These results suggest that the longer half-time of somatostatin-28 in diabetic rats in vivo is due to its slower hepatic clearance. The hepatic clearance of somatostatin-28 and somatostatin-14 was further studied in vitro using a recirculating liver perfusion method. The hepatic clearance of 1.2 nM of either somatostatin-28 or somatostatin-14 was significantly lower in diabetic rats than in controls (p less than 0.01). This indicates that elevated plasma somatostatin levels in diabetic rats are caused at least in part by decreased hepatic clearance of somatostatin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Growth and mitogenic effects of arylphorin in vivo and in vitro

In insects, developmental responses are organ- and tissue-specific. In previous studies of insect midgut cells in primary tissue cultures, growth-promoting and differentiation factors were identified from the growth media, hemolymph, and fat body. Recently, it was determined that the mitogenic effect of a Manduca sexta fat body extract on midgut stem cells of Heliothis virescens was due to the presence of monomeric alpha-arylphorin. Here we report that in primary midgut cell cultures, this same arylphorin stimulates stem cell proliferation in the lepidopterans M. sexta and Spodoptera littoralis, and in the beetle Leptinotarsa decemlineata. Studies using S. littoralis cells confirm that the mitogenic effect is due to free alpha-arylphorin subunits. In addition, feeding artificial diets containing arylphorin increased the growth rates of several insect species. When tested against continuous cell lines, including some with midgut and fat body origins, arylphorin had no effect; however, a cell line derived from Lymantria dispar fat body grew more rapidly in medium containing a chymotryptic digest of arylphorin.     

Antiplatelet and antithrombotic activities of purpurogallin in vitro and in vivo

Enzymatic oxidation of pyrogallol was efficiently transformed to an oxidative product, purpurogallin (PPG). Here, the anticoagulant activities of PPG were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin and activated factor X (FXa). And, the effects of PPG on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-α activated human umbilical vein endothelial cells (HUVECs). Treatment with PPG resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, PPG inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. PPG also elicited anticoagulant effects in mice. In addition, treatment with PPG resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, PPG possesses antithrombotic activities and offers a basis for development of a novel anticoagulant. [BMB Reports 2014; 47(7): 376-381]     

Growth and reproduction of Arabidopsis thaliana in relation to storage of starch and nitrate in the wild-type and in starch-deficient and nitrate-uptake-deficient mutants

We have investigated the interactions between resource assimilation and storage in rosette leaves, and their impact on the growth and reproduction of the annual species Arabidopsis thaliana. The resource balance was experimentally perturbed by changing (i) the external nutrition, by varying the nitrogen supply; (ii) the assimilation and reallocation of resources from rosette leaves to reproductive organs, by cutting or covering rosette leaves at the time of early flower bud formation, and (iii) the internal carbon and nitrogen balance of the plants, by using isogenic mutants either lacking starch formation (PGM mutant) or with reduced nitrate uptake (NU mutant). When plants were grown on high nitrogen, they had higher concentrations of carbohydrates and nitrate in their leaves during the rosette phase than during flowering. However, these storage pools did not significantly contribute to the bulk flow of resources to seeds. The pool size of stored resources in rosette leaves at the onset of seed filling was very low compared to the total amount of carbon and nitrogen needed for seed formation. Instead, the rosette leaves had an important function in the continued assimilation of resources during seed ripening, as shown by the low seed yield of plants whose leaves were covered or cut off. When a key resource became limiting, such as nitrogen in the NU mutants and in plants grown on a low nitrogen supply, stored resources in the rosette leaves (e.g. nitrogen) were remobilized, and made a larger contribution to seed biomass. A change in nutrition resulted in a complete reversal of the plant response: plants shifted from high to low nutrition exhibited a seed yield similar to that of plants grown continuously on a low nitrogen supply, and vice versa. This demonstrates that resource assimilation during the reproductive phase determines seed production. The PGM mutant had a reduced growth rate and a smaller biomass during the rosette phase as a result of changes in respiration caused by a high turnover of soluble sugars ( Caspar et al. 1986 ; W. Schulze et al. 1991 ). During flowering, however, the vegetative growth rate in the PGM mutant increased, and exceeded that of the wild-type. By the end of the flowering stage, the biomass of the PGM mutant did not differ from that of the wild-type. However, in contrast to the wild-type, the PGM mutant maintained a high vegetative growth rate during seed formation, but had a low rate of seed production. These differences in allocation in the PGM mutant result in a significantly lower seed yield in the starchless mutants. This indicates that starch formation is not only an important factor during growth in the rosette phase, but is also important for whole plant allocation during seed formation. The NU mutant resembled the wild-type grown on a low nitrogen supply, except that it unexpectedly showed symptoms of carbohydrate shortage as well as nitrogen deficiency. In all genotypes and treatments, there was a striking correlation between the concentrations of nitrate and organic nitrogen and shoot growth on the one hand, and sucrose concentration and root growth on the other. In addition, nitrate reductase activity (NRA) was correlated with the total carbohydrate concentration: low carbohydrate levels in starchless mutants led to low NRA even at high nitrate supply. Thus the concentrations of stored carbohydrates and nitrate are directly or indirectly involved in regulating allocation.     

Importance of guanine nitration and hydroxylation in DNA in vitro and in vivo

Guanine (Gua) modification by nitrating and hydroxylating systems was investigated in DNA. In isolated calf thymus DNA, 8-NO(2)-Gua and 8-oxo-Gua were dose-dependently formed with peroxynitrite, and 8-NO(2)-Gua was released in substantial amounts. Myeloperoxidase (MPO) with H(2)O(2) and NO(2)(-) reacted with calf thymus DNA to form 8-NO(2)-Gua dose dependently without release of 8-NO(2)-Gua. The frequency of strand breaks was higher than the sum of 8-NO(2)-Gua and 8-oxo-Gua, particularly in the MPO-treated DNA, indicating the importance of other types of damage. The activation of human neutrophils and lymphocytes with phorbol ester did not induce 8-NO(2)-Gua and 8-oxo-Gua in their nuclear DNA. However, 8-NO(2)-Gua was found in calf thymus DNA co-incubated with activated neutrophils in the presence of NO(2)(-). No significant formation of 8-NO(2)-Gua was found in liver DNA from mice treated with Escherichia coli lipopolysaccharide. The incubation of peroxynitrite or MPO-H(2)O(2)-NO(2)(-)-treated DNA with formamidopyrimidine glycosylase (Fpg) released 8-oxo-Gua, but not 8-NO(2)-Gua, indicating that 8-NO(2)-Gua is not a substrate for Fpg. Although 8-NO(2)-Gua was generated in isolated DNA by different nitrating systems, other types of damage were formed in abundance, and the lesion could not be found reliably in nuclear DNA, suggesting that the biological importance is limited.