Uptake and Intracellular Distribution of Oligonucleotides Vectorized by a PAMAM Dendrimer

Abstract

We studied the uptake and intracellular distribution of an FITC labelled phosphodiester oligodeoxynucleotide (ODN) vectorized by a dendrimeric structure in cell culture.     

Enhanced Specificity of Minimally Modified Anti-c-myc Oligonucleotides

Abstract

The sequence-specificity of antisense oligonucleotides (ODN) against c-myc mRNA was tested by Northern blot analysis. Rat smooth muscle cells were treated with antisense or control ODN against c-myc modified by the “minimal protection strategy”. At 0.3 μM concentration the ODN show a very specific reduction in c-myc mRNA levels. Use of the “minimal protection strategy” minimizes nonspecific effects as observed for all-phosphorothioate ODN containing four consecutive guanine residues.     

DcR3蛋白在人食管鳞状细胞癌中的表达及临床意义

目的:探讨诱骗受体(Decoy receptor 3,Dc R3)蛋白在人食管鳞状细胞癌中的表达和临床意义。方法:通过免疫组化的方法,在人食管鳞状细胞癌组织和正常食管粘膜组织各112例中检测Dc R3蛋白的表达差异,并分析Dc R3蛋白表达与食管鳞状细胞癌临床病理学特征间的相关性。结果:在组织标本中检测发现,Dc R3蛋白在食管鳞状细胞癌组织中的阳性表达为72.3%,在正常食管粘膜组织中的阳性表达为25.9%。卡方检验分析表明,Dc R3蛋白在食管鳞状细胞癌组织中的阳性表达与食管鳞状细胞癌的临床分级(P=0.002)、淋巴结转移(P0.001)、组织学分级(P=0.01)具有显著相关性,而与患者的年龄(P=0.673)、性别(P=0.378)、抽烟史(P=0.392)、饮酒史(P=0.093)等均无关。Spearman相关性分析检测发现,Dc R3蛋白高表达与高临床分级(r=-0.213,P=0.024)、高组织学分级(r=-0.285,P=0.002)、淋巴结转移(r=-0.568,0.001)呈正相关,而与食管鳞状细胞癌患者的年龄、性别、吸烟史、饮酒史等均无相关性(P0.05)。结论:Dc R3蛋白在食管鳞状细胞癌组织中表达更高,且与食管鳞状细胞癌疾病的进展、转移呈正相关。Dc R3蛋白表达可用来检测食管鳞状细胞癌疾病严重程度。     

Oral Administration of an Immunostimulatory DNA Sequence from Bifidobacterium longum Improves Th1/Th2 Balance in a Murine Model

We have reported the antiallergic activities of the immunostimulatory oligodeoxynucleotide (ODN) BL07S, identified from genomic DNA of Bifidobacterium longum BB536 from in vitro and in vivo studies. The present study evaluated the efficiency of ODN BL07S in preventing allergic responses by oral administration. Oral administration of BL07S suppressed serum ovalbumin (OVA)-specific immunoglobulin (Ig) E levels and improved the OVA-specific IgG2a/IgG1 ratio. ODN BL07S increased Th1 cytokine and decreased Th2 cytokine production in splenocytes. These results suggest that immunostimulatory ODNs are potentially associated with the antiallergic effects of probiotics.     

Grassflies of the genus <Emphasis Type="Italic">Polyodaspis</Emphasis> Duda (Diptera,Chloropidae) in the fauna of southeast Asia

Over 200 specimens of Polyodaspis from Vietnam and Thailand (project TIGER) were studied. Variations in the color of the body and, especially, of the setae and setulae of the head, scutellum, and abdomen are demonstrated. The following new synonymy is established: Polyodaspis ruficornis (Macquart, 1835) = P. ruficornis var. tarsalis Frey, 1923, syn. n.; = P. flavipila Duda, 1934, syn. n.; = P. endogena de Meijere, 1938, syn. n.; = P. flavisetosa Nartshuk, 1991, syn. n.; = P. similis Nartshuk, 1991, syn. n.     

Disordered Single Crystal Evidence for a Quadruple Helix Formed by Guanosine 5′-Monophosphate

Abstract

The disodium salt of guanosine 5′-monophosphate (5′-GMP) has been crystallized earlier in an orthorhombic array. We have obtained a new crystal form of 5′-GMP at pH 8 which reveals a clear helical nature, with guanine bases stacked perpendicular to the helix axis. Although the X-ray pictures show partial disorder, they can be indexed on a hexagonal net with a = b = 28.6 Å,c = 9.8 Å, V= 6942ų(1Å = 0.1 nm). The probable space group is P6₄, and past experience with ca. 600 ų per base in oligonucleotide crystals suggests that the cell contains 12 GMP molecules. The crystal packing parameters and the intensity distribution agree with a model of three hydrogen-bonded guanine tetrads in the unit cell, stacked so as to build a quadruple helix similar to that proposed earlier from fiber studies (Zimmerman, S.B., J. Mol. Biol. 106, 663–672 (1976)).     

Meiosis,SC-formation,and karyotype structure in diploidPaeonia tenuifolia and tetraploidP. officinalis

The meiosis of the diploidPaeonia tenuifola and the allotetraploidP. officinalis was studied after conventional methanol/acetic acid-fixation and synaptonemal complex (SC) spreading. Meiosis inP. tenuifolia (2n = 10) is normal with five bivalents in metaphase I, and the SCs in pachytene show regular features. InP. officinalis (2n = 4x = 20) univalents, bivalents and multivalents are found in metaphase I. The SCs reveal several abnormalities: a high number of unpaired lateral elements, partner exchanges between three and four lateral elements, loops and lateral element thickenings. These characteristics are compared with the situations found in other polyploid and hybrid species. It is noteworthy that the abnormalities in meiosis ofP. officinalis are not reflected in its somatic karyotype. Its features were analysed after silver staining and fluorescent staining with chromomycin and compared with those ofP. tenuifolia. Synaptonemal Complex Spreading in Plants2; for part1 see Pl. Syst. Evol.154, 129–136 (1986).     

Nickel distribution in the hyperaccumulator Alyssum serpyllifolium Desf. spp. from the Iberian Peninsula

Abstract

The relative concentration and distribution of nickel (Ni) in vegetative tissues (leaves, stems and trichomes) and reproductive organs (seeds) was studied using energy-dispersive X-ray microanalysis (EDXS) and scanning electron microscopy (SEM) in two previously studied Ni-hyperaccumulator subspecies of Alyssum serpyllifolium Desf. growing naturally in ultramafic soils of the Iberian Peninsula: A. serpyllifolium ssp. lusitanicum Dudley & P. Silva and A. serpyllifolium ssp. malacitanum Rivas Goday ex G. López. Both taxa showed that Ni accumulates preferentially in the leaves, exhibiting a homogeneous distribution on both epidermis surfaces. The highest Ni concentrations were found inside the epidermal cells and at the base of trichome stalks. Ni accumulation in seeds was lower than in the vegetative organs. The location of Ni in these plants allows us to postulate that its accumulation is a protection mechanism against external stress.     

Stenogrammitis, a new genus of grammitid ferns segregated from Lellingeria (Polypodiaceae)

Stenogrammitis , a new genus of grammitid ferns, is segregated from Lellingeria based on morphological and molecular evidence. It differs from Lellingeria by linear leaves usually less than 5 mm wide, clathrate iridescent rhizome scales that are glabrous except for a single apical cilium, veins unbranched and only one per segment, fertile veins usually with the dark sclerenchyma visible beneath the sporangia, and x = 33. In contrast, Lellingeria has broader laminae, veins pinnate within the segments, and fertile veins not visible beneath the sporangia. Melpomene, which is sister to Stenogrammitis and Lellingeria, differs from those two genera by reddish setae on the leaves and rhizome scales papillate at the apex. Some species of Stenogrammitis are also distinctive by hemidimorphic laminae that have the fertile portion less dissected than the sterile. Stenogrammitis is pantropical and currently comprises 24 species, 12 of which occur in the Neotropics, six in Africa, four in Madagascar, and two on Pacific Islands. New combinations are made for Stenogrammitis aethiopica, S. anamorphosa, S. ascensionensis, S. boivinii, S. delitescens, S. jamesonii, S. hartii, S. hellwigii, S. hildebrandtii, S. limula, S. luetzelburgii, S. myosuroides, S. nutata, S. oosora, S. paucipinnata, S. prionodes, S. pumila, S. ruglessii, S. rupestris, S. saffordii, S. strangeana, S. tomensis, S. subcoriacea, and S. wittigiana. Lectotypifications are made for Grammitis muscosa, Polypodium itatiayense, P. oosorum var. micropecten, P. serrulatum forma major, P. serrulatum forma minor, S. luetzelburgii, S. myosuroides, and S. wittigiana. Illustrations are presented for the diagnostic characters of the genus, as well as a map with the geographical distribution.     

N-cadherin在卵巢癌中的表达及其临床病理意义

目的:探讨N-cadherin在人类卵巢癌组织中的表达及其临床-病理意义。方法:采用免疫组织化学染色法检测281例卵巢癌患者肿瘤组织中N-cadherin的表达,并分析其与患者临床病理特征之间的关系。结果:N-cadherin在卵巢癌原发灶中的表达显著低于配对的转移灶(P=0.018);卵巢癌组织中N-cadherin的表达与患者FIGO分期(P=0.034)、组织学类型(P0.001)、肿瘤分级(P=0.004)均显著相关。结论:N-cadherin高表达更多见于进展期(FIGOⅡ-Ⅳ)卵巢癌、高级别浆液性癌及高级别卵巢癌,可能与卵巢癌细胞的侵袭及迁移能力呈正相关,对判断卵巢癌的生物学行为具有重要的参考价值。     

茅尾海无瓣海桑内生细菌多样性及其细胞毒活性北大核心CSCD

【目的】对无瓣海桑各组织器官内生细菌的分布特征、物种多样性及细胞毒活性进行分析。【方法】采用稀释涂布法分析无瓣海桑内生细菌的菌落形态及分布。利用16S r RNA基因序列进行系统发育分析,探讨无瓣海桑内生细菌的物种多样性。利用MTT法测试内生细菌培养液的乙酸乙酯提取物细胞毒活性。【结果】从各组织器官中分离出内生细菌38株,隶属12科21属(其中5属未定科),内生细菌数量和群落结构组成存在明显的组织特异性。在分离的菌株中,有5株菌与已有细菌物种典型菌株的全长16S r RNA基因相似性低于97%,代表着潜在新属或新种。5株内生细菌(R74、R71、S92、S85、S84)具有较强的细胞毒活性。【结论】无瓣海桑中可培养内生细菌的物种多样性丰富,潜藏较丰富的新物种资源,且含有较为丰富的活性菌株。     

Convenient Synthesis of Oligodeoxynucleotides Containing 2′-Deoxy-6-thioinosine

Abstract

A facile synthesis of oligodeoxynucleotides (ODN) containing 2′-deoxy-6-thioinosine (dI^6S) based on the convertible nucleoside O6-phenyl-2′-deoxyinosine is presented. After standard solid-phase DNA synthesis and removal of the cyanoethyl protecting groups with DBU treatment with aqueous sodium hydrogen sulfide introduces the sulfur functionality, deprotects the other nucleobases and cleaves the ODN from the solid support in a one-pot reaction. In addition, the extinction coefficient of 2′-deoxy-6-thioinosine is determined by enzymatic fragmentation of the resulting ODN in the presence of adenosine deaminase.     

林下参内生生防细菌的分离鉴定及定殖能力

【背景】多年生林下参在自然环境下生长多年,其体内存在的内生菌具有更强的适应性和定殖性,可以提高植物自身抗性,抑制病原菌的生长,更好地发挥与植物的互作。【目的】筛选定殖能力强、繁殖能力快且对病原菌具有拮抗作用的优势菌株。【方法】采用常规组织分离方法,从健康林下参根部组织中分离内生菌,通过对峙试验筛选出对人参病原菌有拮抗作用的内生细菌并对其以传统的鉴定方法进行鉴定。【结果】在得到的6株内生细菌中,菌株LXS-N2对人参立枯病病原菌、人参猝倒病病原菌均有明显抑菌性,而且具有定殖性好、繁殖快的特点,通过破坏病原真菌细胞壁和细胞膜以及改变菌丝形态从而抑制病原真菌生长。【结论】经形态学观察、生理生化反应及16S rRNA基因序列分析鉴定内生菌LXS-N2为贝莱斯芽孢杆菌,具有良好的应用开发潜力。     

Specifically Designed Polymeric Nanospheres Increase Cellular Uptake of Unmodified Antisense ODNs

Abstract

The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reversibly bound to new polymeric nanoparticles in HL-60 cellular system have been found to increase by 50 folds if compared with the free ODN. An initial single dose (320 nM) of the nanoparticle bound unmodified antimyb ODN has been able to specifically inhibit HL-60 leukemia cell proliferation for at least 8 days.     

Reproductive effort,fecundity and energy allocation in two species of the genus Perinereis (Polychaeta: Nereididae)

Summary

The reproductive effort in terms of fecundity and energy allocation was studied in two species of semelparous polychaetes belonging to the genus Perinereis, living in the same environment, with different reproductive modalities. There is a great individual variability both in terms of reproductive effort and fecundity. Fecundity varied from 4080 to 15000 oocytes in P. rullieri and from 7000 to 26000 in P. cultrifera; no linear relationship was found between oocyte number and total jaw length utilised as size index. The energy content of germinal and somatic tissues was determined by Differential Scanning Calorimeter (DSC). The reproductive effort was calculated as RE = E_G/(E_G + E_S) where E_G is the total energy in germinal tissues and E_S is the total energy in somatic tissues. Reproductive effort is very high with mean values of 0.62 for P. rullieri and 0.79 for P. cultrifera. The different amounts of energy allocated in germinal tissues can be attributed to the different reproductive modalities—P. rullieri reproduces in the atokous phase whereas P. cultrifera has conserved epitoky in its life-cycle. The lack of correlation between reproductive effort and size index strongly suggests that reproductive allocation does not increase with age. In semelparous species the variability in fecundity and reproductive effort observed cannot be interpreted in terms of a trade-off between fecundity and survival as in iteroparous species. In fact, in semelparous an individual allocates all available resources to reproduction and then dies.     

Molecular Structure,Stereochemistry and Interactions of Steffimycin B,and DNA Binding Anthracycline Antibiotic

Abstract

The crystal and molecular structure of anthracycline antibiotic steffimycin B(C₂₉H₃₂₀O₁₃) has been determined by X-ray diffraction and the stereochemistry revealed. The orthorhombic crystals belong to space group P2₁2₁2₁, with the dimensions; a = 8.253 (2), b = 8.198 (2), c = 40.850 (8) Å and Z = 4. Intensity data were collected for 2518 independent reflections. The structure was solved by direct methods and refined to an R value of 0.066 for 1410 reflections. The configuration in ring A is TR,8S,9S. Ring A adopts half chair conformation, while the sugar ring has the regular chair conformation. The molecule most probably binds to double helical DNA through intercalation and hydrogen bonding.     

高海拔地区藏族妇女P21Ser31Arg多态性及与宫颈癌遗传易感性的相关性研究

目的:研究高海拔地区藏族妇女宫颈组织中P21Ser31Arg的多态性,并分析其与宫颈癌遗传易感性的相关性。方法:选择2015年7月~2016年6月收治的200例宫颈癌患者、100例宫颈上皮内瘤样病变(CIN)患者、100例宫颈炎患者及100例正常宫颈患者为研究对象,采用限制性酶切片段长度多态性分析(PCR-RFLP)方法检测各组患者宫颈组织中P21Ser31Arg三种基因型Ser/Ser、Ser/Arg及Arg/Arg的分布情况并比较其构成差异,进一步分析其多态性与宫颈癌患者年龄、癌症分型及分化程度的相关性。结果:四组患者宫颈组织中均检测到三种基因型,Ser/Arg、Ser/Ser及Arg/Arg三种基因型的构成比分别为53.38%、34.88%及11.75%。四组患者宫颈组织中P21Ser31Arg的三种基因型分布整体比较及各组两两比较,差异均无统计学意义(P0.05)。200例宫颈癌患者中,不同年龄组及不同分型组患者的P21Ser31Arg基因型分布比较,差异无统计学意义(P0.05);而不同分化程度组浸润性鳞癌患者的三种基因型分布比较,差异具有统计学意义(P0.05)。结论:未发现P21Ser31Arg多态性与高海拔地区藏族妇女宫颈癌的遗传易感性有关,而可能与宫颈癌患者的分化程度相关。     

Poterium Spinosum L. Elemento Del Mediterraneo Orientale A Calamosca (Capo S. Elia,Sardegna)

Abstract

«Poterium spinosum» L. in Western Mediterranean at Calamosca (Capo S. Elia, Sardegna). — The Author has studied the station of Poterium spinosum at Calamosca, a small valley of S. Elia Cape, near Cagliari, Sardinia. Im Sardinia this place is the only one where P. spinosum grows on the contrary to the information given in RIKLI (1946) where P. spinosum is indicated as growing everywhere along the coast in the island. As it is known, P. spinosum is very common along the coast in Greece, Lebanon, Israel, Syria; is absent in Egypt; is present in the calcareous part of Cyrenaica, in Sicily between Siracusa and Capo Passero; it reaches westwards the small island of Djerba (Tunisia) and Calamosca (S. Elia Cape — Sardinia). In Dalmatia has been reported for Spalato (Split) and Ragusa (Dubrovnik). It has not been found lately at Fiume (Rijeka) neither at Tivoli (Latium), Cotrone (Basilicata) and Bari (Apulia). P. spinosum at Calamosca in the higher part of the station is tipically rock-dwelling, lower is still rock-dwelling but associated with Thymus capitatus H. et Lk., while in the lowert part of the station P. spinosum grows on gravel with Thymelaea hirsuta Endl. P. spinosum at Calamosca has never been found growing with Thymus capitatus and Thymelaea hirsuta both. The center of the distribution area of P. spinosum is in eastern Mediterranean where it forms (Greece and Greek islands) a typical association with Thymelaea hirsuta (RIKLI 1946). In Israel P. spinosum is associated with Thymus capitatus (ZOHARY). The distribution area of P. spinosum is solid in its eastern part, fragmented in central Mediterranean and sparse towards the west. Recently the species, which has 2n = 28 chromosomes (LARSEN 1955), has been found to have hypoglycemic principles active against diabetes.     

Recurrence prediction using microRNA expression in hormone receptor positive breast cancer during tamoxifen treatment

Abstract

Purpose: To identify miRNAs associated with distant recurrence during tamoxifen treatment and build a recurrence prediction model.

Materials and methods: We measured the expression of five miRNAs (miR-134, miR-125b-5P, miRNA-30a, miR-10a-5p and miR-222). A total of 176 tumour tissues from 176 patients who had hormone receptor positive breast cancer with tamoxifen treatment were used to measure miRNA expression using quantitative real-time PCR (qRT-PCR).

Results: The five miRNAs were all up-regulated in distant recurrence cases within 5?years after surgery and during tamoxifen treatment. Kaplan-Meier survival analyses based on expression cut-offs determined by receiver characteristics curves (ROC) showed that high expression of miR-134, miR-125b-5P, miRNA-30a, miR-10a-5p and miR-222 were significantly (log-rank p-value =0.006, p-value <0.0001, p-value <0.0001, p-value <0.0001 and p-value <0.0001, respectively) associated with short relapse-free time. Our results were used to build a combined 3 miRNAs expression model. It could be used to categorize high-risk subset of patients with short relapse-free survival (AUC =0.891, p-value <0.0001).

Conclusions: Distant recurrence during tamoxifen treatment of hormone positive breast cancer might be affected by tamoxifen resistance related miRNAs. Such distant recurrence can be predicted using miRNA measurement.