Ultrastructure of the Spermatozoa of the Yangtze Finless Porpoise (Neophocaena phocaenoides asiaeorientalis)

P>Semen sample was collected from two captive adult Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis) during physical examination. One individual was aged about 9 years with body length 143 cm (total length) and body weight 46.1 kg in 2003. The age of the other was unknown and its body length was 147 cm and body weight was 43 kg in 2004. Ultrastructure of their spermatozoa was examined using scanning and transmission electron microscope. The sperm concentration was 4.17 x 10(9) spermatozoa per ml by the cytometer. The approximate dimensions of the spermatozoa were as follows: head length, 3.366 +/- 0.140 mu m (mean +/- SE, n = 15); head width, 1.896 +/- 0.099 mu m (n = 15); and neck length, 1.004 +/- 0.074 mu m (n = 10). The tail included midpiece, principal piece and terminal piece. The length of the midpiece was 1.882 +/- 0.077 mu m (n = 9). There is no apparent boundary between the principal piece and the terminal piece, so the length of the principal piece and the terminal piece was 44.612 +/- 3.485 mu m (n = 5). Total length of the spermatozoa was 53.314 +/- 4.580 mu m (n = 10). The acrosome covered approximately 45.8% of the anterior portion of the head.

The first non-mammalian CXCR3 in a teleost fish: Gene and expression in blood cells and central nervous system in the grass carp (Ctenopharyngodon idella)

A novel fish chemokine receptor gene, chemokine (C-X-C motif) receptor 3 (CXCR3)-like was isolated from the grass carp Ctenopharyngodon idella , with its full-length genomic sequence. The cDNA of grass carp CXCR3-like (gcCXCR3-like) consists of 1261 bp with a 49bp 5'-UTR and a 189 bp 3'-UTR. An open reading frame of 1023 bp encodes a 341-amino acid peptide, with seven transmembrane helices. The deduced amino acid sequence showed the same sequence identities (37.8%) with its counterparts in goat and human. The gcCXCR3-like gene consists of two exons, with one intervening intron, spaced over approximately 2 kb of genomic sequence. Phylogenetic analyses clearly demonstrated that the gcCXCR3-like resembles the CXCR3s of other vertebrates. Real-time PCR analysis showed that gcCXCR3-like was expressed in all tested organs except heart and the expression level of gcCXCR3-like was highest in brain. Flow cytometric analyses showed the positive rate of labelled leukocytes from the healthy grass carp was 17.3%, and the labelled leukocytes were divided into three types by cell sorting. Immunohistochemical localization revealed that gcCXCR3-like expressed in whole brain regions including cerebel, diencephalon, medulla oblongata, optic lobe, and rhinencephalon, and that the labelled leukocytes are actually populations of monocyte and/or phagocyte, lymphocyte and the granulocyte. It is considered that fish CXCR expression and their function may need to be investigated in both nervous and immune systems. (c) 2006 Elsevier Ltd. All rights reserved.

Preparation of pH-sensitive polymer by thermal initiation polymerization and its application in fluorescence immunoassay

A fluorescence immunoassay for human IgG (Ag) was developed using a pH-sensitive polymer prepared by thermal initiation or redox initiation polymerization as a carrier. In the competitive immunoassay, appropriate quantity of Ag was immobilized on the polymer and the standard Ag (or sample) solution, and a constant amount of fluorescein isothiocyanate labeled goat anti-human IgG antibody (Ab-FITC) was added. Immobilized Ag and the standard (or sample) Ag competed for binding to the Ab-FITC in 37 C in homogeneous format. After changing the pH to separate the polymer-immune complex precipitate, it was re-dissolved and determined by fluorescence method. The results showed that the immobilization efficiency, immunological reaction activities of immobilized Au and phase transition pH range were improved as Ag was immobilized by thermal initiation instead of redox initiation polymerization. Under optimum conditions, the calibration graphs for the Ag in both methods, thermal initiation and redox initiation, were linear over the concentration range of 0.0-1000 ng mL(-1), with detection limits 8 (thermal initiation) and 12 ng mL(1) (redox initiation), respectively. Moreover, some pH-sensitive polymer prepared only in organic solvent or under high temperature could also be used as an immunoreaction carrier by thermal initiation polymerization. Thermal initiation polymerization was a better immobilization mode. (C) 2004 Elsevier B.V. All rights reserved.

黄土丘陵区免耕条件下两种土壤酶活性变化

为了确定合理环保的耕作制度,2007~2008安塞田间定位试验黄土丘陵旱作农区大豆(Glycine maxL)、玉米(Zea maysL)、红小豆(Semen Phaseoli)、马铃薯(Solanum tuberosumLinn.)在翻耕化肥(CF)、翻耕有机肥(CM)、翻耕无肥(CN)、免耕化肥(NF)、免耕有机肥(NM)、免耕无肥(NN)等水平下的农田土壤脲酶、蔗糖酶活性。结果表明:在作物花期,大豆、玉米土壤脲酶活性较高,蔗糖酶活性较低,而红小豆、马铃薯则与之相反,差异极显著。到作物收获后,玉米土壤脲酶、蔗糖酶活性增高,增幅在83%以上,而马铃薯、红小豆、大豆三种作物土壤脲酶活性降低,降幅在10%以上,蔗糖酶活性增强,增幅在40%以上。从花期到收获后,免耕降低了土壤脲酶活性,提高了土壤蔗糖酶活性,在黄土丘陵沟壑旱作农区两种土壤酶活性表现较优的作物为大豆、玉米,较优的处理为NM玉米、NF大豆,其田间环保效应为:作物生长期间两种酶活性比较高,收获后两种酶活性则降低,有利于提高作物生长期土壤肥效利用率,减少作物收获后温室气体的排放。

陕北黄土丘陵沟壑区水土保持耕作及施肥下农田土壤种子库特征

为了探明多年免耕下农田恶性杂草发生的机理,提高保护性耕作下作物对农田恶性杂草持久稳定的抑制效果,依据陕西安塞田间4a的定位试验,采用小区调查取样和室内实验相结合的方法,从物种组成、密度特征、多样性以及相似性特征等方面,研究了黄土丘陵旱作农区大豆(Glycine max)、玉米(Zea mays)、红小豆(Semen Phaseoli)、马铃薯(Solanum tuberosum)在翻耕化肥(CF)、翻耕有机肥(CM)、翻耕无肥(CN)、免耕化肥(NF)、免耕有机肥(NM)、免耕无肥(NN)等水平下的农田土壤种子库。结果表明:(1)4种作物24种土样中共萌发出12个物种1965株幼苗,隶属于7科12属。1年生杂草占94%,棒头草(fugax nees ex steud)、苋菜(Acalypha australis)、马唐(Digitaria sanguinalis)、早熟禾(Poasphondylodes)为优势种,占87%。(2)在0～20cm土层不同处理间,土壤种子库的密度变动于(282.9±63.4)～(7482.5±1078.3)粒.m-2,其中,红小豆小区>马铃薯小区>大豆小区>玉米小区;翻耕小区>免...

Ginsenoside R-e increases fertile and asthenozoospermic infertile human sperm motility by induction of nitric oxide synthase

We investigated the effects of Ginsenoside R-e on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or 100 mu M of Ginsenoside R-e. Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the H-3-arginine to H-3-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside R-e significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside R-e. And pretreatment with a NOS inhibitor N-omega-Nitro-L-arginine methyl ester (L-NAME, 100 mu M) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside R-e. Data suggested that Ginsenoside R-e is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.

Electrochemical Metalloimmunoassay Based on Enlargement of Gold Nanoparticle Label Treated with Ag Enhancement System

A novel sensitive electrochemical immunoassay with colloidal gold as the antibody labeling tag and subsequent signal amplification by silver enhancement is described. Colloidal gold was treated by a light-sensitive silver enhancement system which made silver deposit on the surface of colloidal gold(form Au/Ag core-shell structure), followed by the release of the metallic silver atoms anchored on the antibody by oxidative dissolution of them in an acidic solution and the indirect determination of the dissolved Ag+ ions by anodic stripping voltammetry(ASV) at a carbon fiber microelectrode. The electrochemical signal is directly proportional to the amount of analyte(goat IgG) in the standard or a sample. The method was evaluated by means of a noncompetitive heterogeneous immunoassay of immunoglobulin G(IgG) with a concentration as low as 0.2 ng/ mL. The high performance of the method is related to the sensitive ASV determination of silver(I) at a carbon fiber microelectrode and to the release of a large number of Ag+ ions from each silver shell anchored on the analyte(goat IgG).

Cryopreservation of flounder (Paralichthys olivaceus) sperm with a practical methodology

A simple and convenient protocol for the cryopreservation of the flounder (Paralichthys olivaceus) sperm was established for "on the spot" cryopreservation of large quantities of semen. The use of three cryoprotectants, dimethyl sulphoxide (DMSO), glycerol (Gly) and methanol was tested in the method. The percentage of motile sperm present in semen after it had been frozen and thawed in the presence of DMSO, Gly or methanol was 60.5 +/- 3.6, 79.17 +/- 4.5 and 13.25 +/- 4.7%, respectively. The fertilization rates of this sperm were 67.06 +/- 15.1, 76.20 +/- 10.0 and 44.93 +/- 22.6%, while the hatching rates of eggs fertilized with this sperm were 37.40 +/- 8.3, 48.18 +/- 25.7 and 23.35 +/- 10.8%, respectively. It was found that Gly and DMSO were better cryoprotectants than methanol, with Gly giving the best overall results. Under scanning electron microscopy, it could be seen that while the majority of the frozen-thawed sperm remained morphologically normal, some exhibited lost or dilated mitochondria, swollen mid-pieces, broken tails, or damaged cell membrane, which probably caused the decrease in motility and fertility of the frozen-thawed sperm. (C) 2003 Elsevier Science Inc. All rights reserved.