基于线粒体CO Ⅰ和Cyt b基因的粉蝶亚科及黄粉蝶亚科(粉蝶科)部分类群的分子系统发生

测定了粉蝶科的粉蝶亚科和黄粉蝶亚科14属共24种线粒体COⅠ和Cyt b基因部分序列,并从GenBank中下载了2种粉蝶的同源序列,以眼蝶科的2个物种为外类群,运用NJ法、贝叶斯法分别重建了分子系统树,探讨了它们的系统发生关系。基因序列分析结果表明,经比对和处理后的序列总长度为1111bp,其中变异位点478个,简约位点382个,碱基T、C、A、G平均含量为39.9%、16.9%、30.9%、12.3%,A＋T含量和C＋G含量分别为70.8%和29.2%。分子系统树显示：黄粉蝶亚科不是单系群,但其中迁粉蝶属和豆粉蝶属在不同的分析方法中均聚合在一起。粉蝶亚科形成一个独立的支系,其中,襟粉蝶族为并系群;粉蝶族的粉蝶属、飞龙粉蝶属和云粉蝶属具有较近的亲缘关系。     

基于ITS-1基因的菜粉蝶地理种群遗传分化研究

为了探讨我国不同地区菜粉蝶种群之间的遗传分化情况, 本研究运用克隆测序法, 对我国16个地区79只菜粉蝶Pieris rapae rDNA的ITS-1基因序列进行测定并分析; 同时, 以东方菜粉蝶Pieris canidia为外群, 重建了它们的系统发生树, 初步探讨了它们的生物地理学分化格局。序列分析结果显示: 所测菜粉蝶的ITS-1序列长度介于337~458 bp之间。经序列比对后, 在347个位点中共有281个保守位点, 63个变异位点, 16个简约信息位点和47个单突变位点; AMOVA软件分析其核苷酸多态性指数(nucleotide diversity) Pi为0.014, 每位点Theta (per site) Eta为0.041, 遗传分化指数(F_ST)为0.351 (P<0.05); DnaSP软件共检测出48个单倍型(haplotype)序列, 单倍型多样性(haplotype diversity)的频率为0.989。系统发生分析结果表明: 现存的菜粉蝶各地理种群与其地理分布没有直接的对应关系根据系统发生树结果推测, 该蝶种在我国的最早建群可能发生于辽宁、内蒙古及北京一带, 其后, 通过幼虫寄主植物的水陆运输以及成虫的迁飞向我国的不同地区扩散。     

MicroRNA-411a-3p靶向钙/钙调素依赖蛋白激酶4调控羊驼黑色素细胞中的黑色素生成

羊驼是重要的毛用型经济动物,具有22种天然色。毛色由皮肤中黑色素细胞产生的黑色素颗粒的分布和转运所决定的。然而,羊驼色素形成或毛色形成的分子机制复杂,由多个基因、miRNA 和lncRNA调控。但是,miR-411a-3p 对羊驼色素形成起调控作用未见报道。为研究miR-411a-3p在黑色素产生过程中的调控作用,本文将构建的miR-411a-3p载体转染到羊驼黑色素细胞中,并对毛色基因的表达进行研究。经生物信息学预测,钙/钙调素依赖蛋白激酶4(calcium-calmodulin dependent protein kinase 4, CaMK4)可能是miR-411a-3p的靶基因之一。双荧光素酶报告基因结果显示,与对照组相比,双荧光报告酶活性下降（34.78 ± 16.09）%(P<0.01),其下降趋势明显,说明CaMK4可能是miR-411a-3p的靶基因之一。miR-411a-3p通过结合CaMK4的3′ untranslated region (3′-UTR)直接调控CaMK4的表达。在羊驼黑色素细胞中转染miR-411a-3p后,CaMK4、CDK5和TYRP1在转录水平的表达量与NC组相比具有显著下降趋势,其中TYRP1下降趋势尤为显著（53.66 ± 2.11）%（P<0.01）。Western印迹检测CaMK4、CDK5、TYRP1、p-CREB在蛋白质水平的表达与NC组相比下降趋势明显,尤其是CDK5和p-CREB基因下降极为显著,分别为（70.26 ± 4.84）%（P<0.01）和（70.11 ±9.05）%（P<0.01）。Masson-Fontana法检测黑色素颗粒,结果显示,miR-411a-3p抑制黑色素细胞产生黑色素颗粒。通过紫外分光度法检测真黑素（eumelanin,EM）和褐黑素（phenomelanin,PM）显示,EM和PM的含量均被下调。结果表明,miR-411a-3p靶向抑制CaMK4表达,从而改变CREB的表达以控制黑色素的产生,此研究结果对哺乳动物毛色形成机制有重要意义。     

一氧化氮的传递及其对缺氧时呼吸变化的调控

一氧化氮(NO)在体内的传递并非以往认为的随机弥散,而是与血红蛋白(Hb)β亚单位中半胱氨酸的硫醇基(巯基)或其他含巯基化合物相结合,形成亚硝基硫醇(SNOs)。SNOs既具备NO的生物活性又保持其稳定性。SNOs从细胞外进入到胞质的途径是通过酶促反应,而从红细胞输出则经阴离子交换蛋白转运。在缺氧情况下,SNOs对呼吸的调控包括：Hb在肺部的氧合作用,将氧输送到组织和在脑干孤束核调控中枢性呼吸。     

Combination of Active Components of Xiexin Decoction Ameliorates Renal Fibrosis Through the Inhibition of NF-κB and TGF-β1/Smad Pathways in db/db Diabetic Mice

Xiexin decoction, a herbal therapeutic agent commonly used in traditional Chinese medicine, is recognized for its beneficial effects on diabetic nephropathy exerted through the combined action of multiple components, including Rhizoma Coptidis alkaloids (A), Radix et Rhizoma Rhei polysaccharides (P), and Radix Scutellaria flavones (F). Our previous studies have shown that a combination of A, P, and F (APF) exhibits renoprotective effects against diabetic nephropathy. This study was aimed at determining the effects of APF on renal fibrosis in diabetic nephropathy and elucidating the underlying molecular mechanisms. To evaluate the effects of APF, in vivo, db/db diabetic mice were orally administered a low or high dose of APF (300 or 600 mg/kg, respectively) once a day for 8 weeks. We evaluated the blood and urine indices of metabolic and renal function, renal tissue histopathology, renal inflammation, and fibrosis. APF treatment significantly ameliorated glucose and lipid metabolism dysfunction, decreased urinary albumin excretion, normalized creatinine clearance, and reduced the morphological changes in renal tissue. Additionally, APF administration in db/db diabetic mice reduced the elevated levels of renal inflammation mediators such as intercellular adhesion molecule-1, monocyte chemotactic protein-1, tumor necrosis factor-α, interleukin-1β, and active nuclear factor κB (NF-κB). APF treatment also reduced type I and IV collagen, transforming growth factor-β1 (TGF-β1), and TGF-β1 type II receptor expression levels, and decreased the phosphorylation of Smad2/3 in the kidneys of db/db diabetic mice. These results suggest that APF reduces renal fibrosis in diabetic nephropathy through the NF-κB and TGF-β1/Smad signaling pathways. In vitro, APF treatment reduced cell proliferation and protein expression of α-smooth muscle actin, collagen I, TGF-β1 and NF-κB in mesangial cells cultured with high glucose concentrations. Our findings indicate that treatment with multi-component herbal therapeutic formulations may be a useful approach for the treatment of diabetic nephropathy.     

Analysis of genetic diversity on 9 wild stocks of taimen (Hucho taimen) by microsatellite markers

Taimen (Hucho taimen) is a native fish species in China and it is in the state of endangerment. To explain clearly the genetic diversity and genetic structure, 9 wild populations of taimen were investigated using 20 microsatellite markers. The results showed that their observed heterozygosity ranged from 0.0994 to 0.8882, the expected heterozygosity varied from 0.2005 to 0.8759, and the range of PIC index was from 0.3432 to 0.5261 while population from Huma River had low genetic diversity. Fst of matching group ranged from 0.0246 to 0.2333 (P <0.0001)and Nm varied among 0.8216 to 9.9292, which indicated that the genetic differentiation was remarkable among populations.The half/full-sib family tests detected a proportion of half/full-sib family groups varying among 27.78% to 90.91%, showing a high inbred pressure and a risk of bottlenecks experienced by most groups. The AMOVA results showed that the global Fst was 0.1081; the clustering result showed that individuals from Beiji tributary of Heilongjiang River clustered as one clade, all individuals from Huma River and Wusuli River clustered as one clade and all individuals from the upper reaches of the Heilongjiang River clustered as another clade. All these results indicated that the decrease of taimen resource has affected the gene exchange among their populations. In order to achieve full protection of taimen germplasm resources, we should put an end to the destructive fishing for taimen and promotegene exchange among their populations.     

大卫绢蛱蝶线粒体基因组全序列测定和分析

目前有关蝶类线粒体基因组全序列及其分子进化的研究报道还不多见。本文利用long PCR和引物步移法得到大卫绢蛱蝶Calinaga davidis的线粒体基因组全序列, 同时就其基因组成和结构特点作了初步分析。结果显示： 其基因组全长为15 267 bp (GenBank登录号为HQ658143), 包括13个蛋白质编码基因(ATP6, ATP8, COI-III, ND1-6, ND4L, Cytb)、22个tRNA基因、2个rRNA基因(16S和12S)以及非编码的控制区。与其他鳞翅目昆虫相一致, 其基因组未出现基因重排现象。基因组共包含11个基因间隔区,总长度为130 bp, 间隔长度1~46 bp, 最大间隔在tRNA^Gln与ND2基因之间; 基因间共存在13处重叠, 总长度为66 bp, 重叠碱基数1~35 bp, 最长的重叠区位于COII与tRNA^Lys基因。lrRNA和srRNA基因长度分别为1 337 bp和773 bp; 除tRNA^Ser(AGN)缺少二氢尿嘧啶臂(DHU stem), 在相应的位置上只形成一个简单环外, 其余的tRNA基因都能形成典型的三叶草结构。13个蛋白编码基因总长度为11 247 bp, 共有3 737个密码子, 它们的碱基组成和密码子的使用具有明显的偏倚性; 除COI外(起始密码子TTG), 其余的12个蛋白质编码基因都以标准的ATN作为起始密码子; COI基因终止密码子为不完全T, ND4基因终止密码子为不完全TA, 其余基因都以TAA为终止密码子。A+T丰富区全长为389 bp, A+T含量高达92.0%, 其中存在2段类似微卫星的重复序列(TA)₆和(AAT)₄。本文的研究结果为探讨绢蛱蝶亚科在蛱蝶科中的系统学地位及其与其他亚科间的系统发生关系等问题提供了重要的分子生物学数据。     

Imidazo[4,5-b]pyridines as corticotropin releasing factor receptor ligands

A series of high affinity CRF receptor ligands with an imidazo[4,5-b]pyridine core is described. Individual analogues were synthesized and tested in a rat CRF receptor binding assay. The best compounds were further tested in the dog N-in-1 pharmacokinetic model to assess plasma levels at 1mg/kg (po) and in the rat situational anxiety model to assess anxiolytic efficacy at 3mg/kg (po). The structure-activity relationships for good receptor binding affinity are described herein.     

基于线粒体COⅠ基因序列分析宝贝科主要类群的系统发生关系

新测了分布于我国海南沿海的11种宝贝科软体动物的线粒体COⅠ基因序列,结合GenBank中17种该科动物的同源性序列片段,基于不同的数学模型,以邻接法、最大简约法和贝叶斯法构建了宝贝科分子系统树.结果表明:卵宝贝亚科Cypraeovulinae是一个单系群;宝贝亚科Cypraeinae和鼻贝亚科Nariinae并非姊妹群,其中,宝贝亚科明显为多系发生,而鼻贝亚科中除疹贝属而外的其他类群可能为单系发生;所有系统树均支持形态学上焦掌贝、绶贝、眼球贝、葡萄贝以及疹贝的单系性及属级分类阶元的界定.鉴于疹贝属在系统树中的特殊位置,建议将其归于宝贝亚科;而龟甲贝和林希那贝间的系统发生关系及其分类地位还有待于进一步探讨.     

Subtype-specific coupling with ADP-ribosyl cyclase of metabotropic glutamate receptors in retina,cervical superior ganglion and NG108-15 cells

Cyclic ADP-ribose (cADP-ribose) is a putative second messenger or modulator. However, the role of cADP-ribose in the downstream signals of the metabotropic glutamate receptors (mGluRs) is unclear. Here, we show that glutamate stimulates ADP-ribosyl cyclase activity in rat or mouse crude membranes of retina via group III mGluRs or in superior cervical ganglion via group I mGluRs. The retina of mGluR6-deficient mice showed no increase in the ADP-ribosyl cyclase level in response to glutamate. GTP enhanced the initial rate of basal and glutamate-stimulated cyclase activity. GTP-gamma-S also stimulated basal activity. To determine whether the coupling mode of mGluRs to ADP-ribosyl cyclase is a feature common to individual cloned mGluRs, we expressed each mGluR subtype in NG108-15 neuroblastoma x glioma hybrid cells. The glutamate-induced stimulation of the cyclase occurs preferentially in NG108-15 cells over-expressing mGluRs1, 3, 5, and 6. Cells expressing mGluR2 or mGluRs4 and 7 exhibit inhibition or no coupling, respectively. Glutamate-induced activation or inhibition of the cyclase activity was eliminated after pre-treatment with cholera or pertussis toxin, respectively. Thus, the subtype-specific coupling of mGluRs to ADP-ribosyl cyclase via G proteins suggests that some glutamate-evoked neuronal functions are mediated by cADP-ribose.