精制木醋液的安全性评价

目的:通过观察实验动物使用木醋液后的急、慢性毒性反应,评估精制木醋液的安全性.方法:①昆明种小鼠,雌雄各10只,木醋液经口灌胃1次,观察1周,并记录动物急性中毒症状及死亡情况.②Wistar大鼠80只,分为受试物高、中、低剂量组和空白对照组,经口灌胃30 d后,观察大鼠血液学、血液生化学及心、肝、脾、肺、肾、胃等主要脏器病理变化.结果:①1周内小鼠未见明显中毒症状及死亡发生,最大耐受剂量为20 g/kg.②30 d后大剂量组血清ALT明显升高,包括对照组偶见肝细胞脂肪变性或糖源变性,但各组间没有显著性差异,其他血液学、生化学、脏器重量和病理检查等无明显变化.结论:本品除了大剂量组出现轻微的生化毒性外,没有发现与受试物有关的其他毒性变化.     

Production of red pigments byMonascus purpureus in submerged culture

For the purpose of mass producingMonascus red pigments optimum medium composition and environmental conditions were investigated in submerged flask cultures. The optimum carbon and nitrogen sources were determined to be 30 g/L of glucose and 1.5 g/L of monosodium glutamate (MSG). Of the three metals examined, Fe²⁺ showed the stronges stimulatory effect on pigment production and some stimulatory effect was also found in Mn²⁺. Optimum pH and agitation speed were determined to be 6.5 and 700 rpm, respectively. Under the optimum culture conditions batch fermentation showed that the maximum biomass yield and specific productivity of red pigments were 0.20 g DCW/g glucose and, 32.5 OD₅₀₀ g DCW⁻¹ h⁻¹, respectively.     

Analysis of proteinase-activated receptor 2 and TLR4 signal transduction: a novel paradigm for receptor cooperativity

Proteinase-activated receptor 2 (PAR(2)), a seven-transmembrane G protein-coupled receptor, is activated at inflammatory sites by proteolytic cleavage of its extracellular N terminus by trypsin-like enzymes, exposing a tethered, receptor-activating ligand. Synthetic agonist peptides (AP) that share the tethered ligand sequence also activate PAR(2), often measured by Ca(2+) release. PAR(2) contributes to inflammation through activation of NF-kappaB-regulated genes; however, the mechanism by which this occurs is unknown. Overexpression of human PAR(2) in HEK293T cells resulted in concentration-dependent, PAR(2) AP-inducible NF-kappaB reporter activation that was protein synthesis-independent, yet blocked by inhibitors that uncouple G(i) proteins or sequester intracellular Ca(2+). Because previous studies described synergistic PAR(2)- and TLR4-mediated cytokine production, we hypothesized that PAR(2) and TLR4 might interact at the level of signaling. In the absence of TLR4, PAR(2)-induced NF-kappaB activity was inhibited by dominant negative (DN)-TRIF or DN-TRAM constructs, but not by DN-MyD88, findings confirmed using cell-permeable, adapter-specific BB loop blocking peptides. Co-expression of TLR4/MD-2/CD14 with PAR(2) in HEK293T cells led to a synergistic increase in AP-induced NF-kappaB signaling that was MyD88-dependent and required a functional TLR4, despite the fact that AP exhibited no TLR4 agonist activity. Co-immunoprecipitation of PAR(2) and TLR4 revealed a physical association that was AP-dependent. The response to AP or lipopolysaccharide was significantly diminished in TLR4(-/-) and PAR (-/-)(2) macrophages, respectively, and SW620 colonic epithelial cells exhibited synergistic responses to co-stimulation with AP and lipopolysaccharide. Our data suggest a unique interaction between two distinct innate immune response receptors and support a novel paradigm of receptor cooperativity in inflammatory responses.     

Fine mapping and candidate gene analysis of <Emphasis Type="Italic">hwh1</Emphasis> and <Emphasis Type="Italic">hwh2</Emphasis>, a set of complementary genes controlling hybrid breakdown in rice

Hybrid breakdown (HB), a phenomenon of reduced viability or fertility accompanied with retarded growth in hybrid progenies, often arises in the offspring of intersubspecific hybrids between indica and japonica in rice. We detected HB plants in F₈ recombinant inbred lines derived from the cross between an indica variety, Milyang 23, and a japonica variety, Tong 88-7. HB plants showed retarded growth, with fewer tillers and spikelets. Genetic analysis revealed that HB was controlled by the complementary action of two recessive genes, hwh1 and hwh2, originating from each of both parents, which were fine-mapped on the short arm of chromosome 2 and on the near centromere region of the long arm of chromosome 11, respectively. A comparison of the sequences of candidate genes among both parents and HB plants revealed that hwh1 encoded a putative glucose-methanol-choline oxidoreductase with one amino acid change compared to Hwh1 and that hwh2 probably encoded a putative hexose transporter with a six amino acid insertion compared to Hwh2. Investigation of the distribution of these alleles among 54 japonica and indica cultivars using candidate gene-based markers suggested that the two loci might be involved in developing reproductive barriers between two subspecies.     

Requirement of multiple protein domains and residues for gating K(ATP) channels by intracellular pH.

ATP-sensitive K(+) channels (K(ATP)) are regulated by pH in addition to ATP, ADP, and phospholipids. In the study we found evidence for the molecular basis of gating the cloned K(ATP) by intracellular protons. Systematic constructions of chimerical Kir6.2-Kir1.1 channels indicated that full pH sensitivity required the N terminus, C terminus, and M2 region. Three amino acid residues were identified in these protein domains, which are Thr-71 in the N terminus, Cys-166 in the M2 region, and His-175 in the C terminus. Mutation of any of them to their counterpart residues in Kir1.1 was sufficient to completely eliminate the pH sensitivity. Creation of these residues rendered the mutant channels clear pH-dependent activation. Thus, critical players in gating K(ATP) by protons are demonstrated. The pH sensitivity enables the K(ATP) to regulate cell excitability in a number of physiological and pathophysiological conditions when pH is low but ATP concentration is normal.     

转化生长因子β及其生物学效应

转化生长因子β（ＴＧＦβ）属于一处多功能多肽家族,在免疫抑制、抗炎症反应、对增殖和细胞间质产生的调控、损伤修复等方面起重要作用。ＴＧＦβ表达和信号转导失常与多处疾病的发生有关。     

Bacteriostatic activities of monoacyl sugar alcohols against thermophilic sporeformers

The bacteriostatic activities of monoacyl sugar alcohols with different acyl chains and hydrophilic heads were examined against some thermophilic sporeformers. Monomyristoyl erythritol and xylitol efficaciously suppressed their spore development. The number and orientation of the hydroxyl groups also played important roles in this activity, and monomyristoyl xylitol exhibited the highest activity among the monomyristoyl sugar alcohols.     

Pacemaker currents modulated by C-type natriuretic peptide in interstitial cells of Cajal from murine small intestine

Although the presence of C-type natriuretic peptide (CNP) in gastrointestinal tract has been demonstrated, the effect of CNP on interstitial cells of Cajal (ICC), pacemaker cells in gastrointestinal tract, is still unclear. This study was designed to investigate the effect of CNP on pacemaker currents of ICC and possible mechanisms. We used immunocytochemistry techniques to exhibit natriuretic peptide receptors (NPR) and recorded membrane currents by using whole-cell patch clamp technique on cultured ICC. Our experiment showed that NPR-A and NPR-B were expressed in ICC from murine small intestine. Whole cell recordings further showed that the amplitude of pacemaker currents in intestinal small networks of ICC was 322+/-22pA and the frequency was 16.25+/-0.95Hz. CNP significantly reduced the amplitude of pacemaker currents in small networks of ICC in a dose-dependent manner, and the amplitude was inhibited by 23.95%, 61.76% and 81.67%, the amplitude values in 329+/-28.0pA, 311.2+/-14.8pA and 295+/-26.5pA before treatment with CNP and 237.9+/-27.5pA, 119.6+/-18.5pA and 57.2+/-13.5pA after treatment with 0.01 micromolxL(-1), 0.1 micromolxL(-1) and 1pmolxL(-1) CNP, respectively. The frequencies of pacemaker currents were also significantly reduced from 16.25+/-0.95Hz of control to 13+/-0.9Hz, 12+/-0.8Hz and 3+/-0.2Hz by 0.01micromolxL 1, 0.1micromolxL(-1) and 1 micromol x L(-1) CNP, respectively. CNP also inhibited the amplitude of pacemaker currents in single ICC. The inhibitory effect of CNP was mimicked by 8-Br-cGMP, a membrane permeable cGMP analogue, which suggests that CNP could inhibit pacemaker currents via NPR-B-particulate guanylate cyclase (pGC)-cGMP signal pathway.     

共聚焦镜观察凋亡巨噬细胞内pH的变化

用透射电镜观察巨噬细胞的形态学改变,结果显示,地塞米松处理８小时后,大部分巨噬细胞发生凋亡特征变化：胞突缩短、减少,胞膜完整。胞体皱缩,胞质密度增加,其中出现大量空泡。胞核染色质边聚、浓缩。另外用激光扫描共聚焦显微镜（ＡＣＡＳ５７０）和ｐＨ荧光探针ＳＮＡＲＦ┐１／ＡＭ实时检测地塞米松处理巨噬细胞胞浆ｐＨ的动态变化。加入地塞米松,多数巨噬细胞胞浆马上发生快速和短期的碱化。随后,胞浆ｐＨ缓慢降低,胞浆酸化。结果表明,胞浆酸化是细胞凋亡发展的必然过程,胞浆碱化则很可能与细胞凋亡的发生相关,也可能与细胞种类、细胞功能状态相关     

Two 17β-hydroxysteroid dehydrogenases (17HSDs) of estradiol biosynthesis: 17HSD type 1 and type 7

Two 17β-hydroxysteroid dehydrogenases (17HSDs), type 1 and type 7, are enzymes of estradiol biosynthesis, in addition to which rodent type 1 enzymes are also able to catalyze androgens. Both of the 17HSDs are abundantly expressed in ovaries, the type 1 enzyme in granulosa cells and type 7 in luteinized cells. The expression of 17HSD7, which has also been described as a prolactin receptor-associated protein (PRAP), is particularly up-regulated in corpus luteum during the second half of rodent pregnancy. A moderate or slight signal for mouse 17HSD7/PRAP mRNA has also been demonstrated in samples of placenta and mammary gland, for example. Human, but not rodent, 17HSD1 is expressed in placenta, breast epithelium and endometrium in addition to ovaries. A cell-specific enhancer, silencer and promoter in the hHSD17B1 gene participate in the regulation of type 1 enzyme expression. The enhancer consists of several subunits, including a retinoic acid response element, the silencer has a binding motif for GATA factors, and the proximal promoter contains adjacent and competing AP-2 and Sp binding sites.