天然丝素制备三维多孔支架

研究盐沥滤法制备三维多孔丝素支架,在丝素多孔支架上培养人成骨肉瘤细胞、成纤维细胞及肝细胞.研究材料的细胞相容性,发现多种动物细胞在盐沥滤法所制备的丝素多孔支架材料上能够很好地黏附和增殖.通过调整丝素水溶液的浓度、溶解丝素溶剂体系、NaCl添加量和NaCl粒径,制备出结构和性质可以调控的三维支架.该结果为丝素多孔支架材料的制备提供了新的研究思路和方法.     

Fluorescence Tracking of Exogenous DNA in Genetic Transformation of the Chinese Oak Silkmoth Antheraea Pernyi via Sperm-Mediated Gene Transfer

Exogenous DNA expressing green fluorescent protein（ GFP） and labeled with fluorescein isothiocyanate（ FITC） was used to transform the Chinese oak silkmoth Antheraea pernyi（ A. pernyi）via sperm-mediated gene transfer（ SMGT）. Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.     

丝素在NMMO·H2O中的溶解及溶液流变性能的研究

探讨了丝素在N-甲基吗啉-N-氧化物一水合物(NMMO.H2O)中的溶解行为,结果发现:天然蚕丝纤维脱胶后得到的丝纤维可以直接溶解在NMMO.H2O中,但是很难得到高于10%浓度的溶液,因而无法用于纺丝.而由再生丝素水溶液制得的再生丝素膜则易于溶解在NMMO.H2O中,并可以得到浓度高达25%的再生丝素/NMMO.H2O溶液,在上述研究的基础上,本文还对再生丝素/NMMO.H2O的流变行为进行了初步研究.     

亲环素A抗原表位在三维结构中的初步定位

以抗人亲环素A单克隆抗体D4作捕获抗体，用噬菌体展示库鉴定出亲环素A模拟抗原表位的氨基酸序列为WSLQSFL，在亲环素A的一级结构中没有相同的序列，提示亲环素A抗原表位为构象型的，亲环素A的三维空间结构已测定，利用RasMol等蛋白质三维结构观察软件，可以初步确定亲环素A的抗原表位的时间位置，此抗原表位与环孢素的结合位点有重叠。     

Cloning of a novel gene associated with human nasopharyngeal carcinoma

One EST N27741 with high expression in normal adult nasopharynx tissues but low expression in adult poorly differentiated squamous nasopharyngeal carcinoma has been selected out by the high-density cDNA array expression profiling technique. The differential expression has been confirmed by RT-PCR. One novel gene of 1096 bp has been cloned based on this EST. Bioinformatics analysis found that the new gene sequence contains a whole reading frame encoding 256 amino acids. There is a stop codon TAA in front of the 5′ end start codon, and a tailing signal AATAAA and poly A tail at the 3′ end. There is no homologous known gene found after searching by blasting this sequence to non-redundancy nucleotide database. Therefore it is considered a novel gene related to nasopharyngeal carcinoma.     

Nanoscale control of silks for regular hydroxyapatite formation

The control of nanoparticle shape is still a challenge for different hydroxyapatite (HA) preparation ways.In the present study,silk fibroin with regular nanostructures was firstly prepared and then emp...     

A point mutation of the rhodopsin gene in one form of retinitis pigmentosa

The gene for autosomal dominant retinitis pigmentosa in a large pedigree of Irish origin has recently been found to be linked to an anonymous polymorphic sequence, D3S47 (C17), from the long arm of chromosome 3. As the gene coding for rhodopsin is also assigned to the long arm of chromosome 3 and is expressed in rod photoreceptors that are affected early in this blinding disease, we searched for a mutation of the rhodopsin gene in patients with autosomal dominant retinitis pigmentosa. We found a C----A transversion in codon 23 (corresponding to a proline----histidine substitution) in 17 of 148 unrelated patients and not in any of 102 unaffected individuals. This result, coupled with the fact that the proline normally present at position 23 is highly conserved among the opsins and related G-protein receptors, indicates that this mutation could be the cause of one form of autosomal dominant retinitis pigmentosa.     

Mutations analysis of STK11 ene in Chinese families with Peutz-Jeghers syndrome

Peutz-Jeghers syndrome (PJS) is an autosomal dominantly inherited disease characterized by multiple gastrointestinal hamartomatous polyps and melanin spots on lips and buccal mucosa, with an increased risk for various cancers. ThePJS gene, a potential tumour suppressor gene, encoding a serine/ threonie kinase (STK11), was mapped to chromosome 19p13.3. To investigate the mutations of STK11 gene in Chinese with PJS, we analyzed its coding sequence in fifteen patientsand twenty unaffected members of six families, including three multigenerational families with PJS and three sporadic families with PJS, by PCR, PCR-DHPLC and DNA sequencing techniques. Ten point mutations were found in the six families, including five missense mutations, one acceptor-splice site mutation, a nonsense mutation and three silent mutations. Our data showed that five missense mutations occurrd at codon 123 (CAG to CAT) in exon 2, codon 161 (ATT to AGT) in exon 4,codon 194 (GAC to GAG) in exon 4, codon 245 (CTC to TTC) in exon 5 and codon 354 (TTC to TTG) in exon 8. One kind of nonsense mutation was detected at codon 37(CAG to TAG) in exon 1. Furthermore, we found an intronic mutation at a splice-acceptor site: a one base substitution from AG to AA in intron 4. These mutations were not detected in 20 normal DNA samples. In three sporadic families, only in one patient, we detected a missense mutation in exon 5. In addition, we found three silent mutations, which may cause polymorphisms of STK11 gene in introns 1(+36), 3(-51) and 5(+27). These results indicated that the point mutation in STK11 might be involved in PJS pathogenesis. Mutation frequency is higher in the families suffering PJS in three or more generations than that of the sporadic cases.     

用信息论方法研究几类核酸序列的密码子使用与基因表达水平的关系

研究了Ｅｓｃｈｅｒｉｃｈｉａｃｏｌｉ，Ｂａｃｉｌｌｕｓｓｕｂｔｉｌｉｓ和ｓａｃｃｈａｒｏｍｙｃｅｓＹｅａｓｔ三类核酸序列的密码子使用频率与基因表达水平的关系，依照每一基因中最适密码子、非最适密码子和稀有密码子出现的概率，计算其信息熵Ｅ。首先，发现Ｅ与ＣＡＩ值保持了很好的线性关系，且三类序列的线性拟合方程非常接近，因此，用信息熵Ｅ作为描述基因表达水平的物理量是合适的；其次，三类序列的Ｅ─Ｐ（Ｒ）（稀有密码子概率）曲线都接近对数关系，说明稀有密码子在抑制基因表达水平方面，起着很重要的作用；第三，由各种关系曲线的相似性，可以推测，Ｅ．ｃｏｌｉ等原核生物与真核生物Ｙｅａｓｔ在密码子使用与基因表达水平的关系方面，其机理是相似的。     

Mutations analysis of STK_(11) gene in Chinese families with Peutz-Jeghers syndrome

Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease that is characterized by multiple gastrointestinal hamartomatous polyps and melanin spots on lips and buccal mucosa at young age[1,2]. Previous studies have demonstrated that PJS predisposes carriers to cancers of gastrointestinal tract, uterus, ovary, testis, breast and other extragastrointestinal organs[3—5]. The STK11 gene, encoding a serine/threonine kinase at chro-mosome 19p13.3, was identified in 1998 as the main causativ…     

Rationalization and Mechanisms of Pulsed Laser Deposition of Fibroin Nanofilms by Using Autogenous Binder

1 Results Homogeneity of fibroin thin films by pulsed laser deposition (PLD) was significantly improved by introducing autogenous binders to consolidate and homogenize the irradiation targets.Homogenization includes among others,decreases in the size of nanostructural units and the fractional debris.Simultaneously,the deposition rate was substantially increased.Autogenous binders are prepared via degumming and liquefaction of fibroin by using Na2CO3 and LiBr,respectively.We also found that optically hom...     

Cu（Ⅱ） effect on the conformation of regenerated silk fibroin in dilute aqueous solution

Much attention has been paid to the natural mechanism of silkworm spinning due to the impressive mechanical properties of the natural fibers. In this work, we studied the effect of Cu（Ⅱ） ions on the secondary structure of Bombyx mori regenerated silk fibroin （SF） in dilute solution by circular dichroism （CD）. The results indicate that a given amount of Cu（Ⅱ） induces the SF conformational transition from random coil to β-sheet, however, further addition of Cu（Ⅱ） is unfavorable for this conversion. Meanwhile, the conformational changes induced by Cu（Ⅱ） follow a nucleation-dependent aggregation mechanism, which is similar to that found in Prion protein （PrP） denaturation and Aβ-peptide aggregations, leading to the neurodegenerative disease. This work would help one understand further the natural spinning process of silkworm. Additionally, it would be significant for the study of the nervous system diseases, because silk fibroin, extracted in large amounts from Bombyx mori silkworm gland, could be a proper model to study PrP denaturation and Aβ-peptide aggregations.     

分离与克隆中国人TPO基因片段及序列分析

应用ＲＴＰＣＲ技术从中国人胎肝细胞中分离出１个５６６ｂｐ大小的基因片段,经过克隆、限制性内切酶鉴定和序列分析证实为ＴＰＯ的ｃＤＮＡ片段．与ＧｅｎＢａｎｋ中发表的人ＴＰＯｍＲＮＡ的序列比较同源性在８２％～９９％之间,仅有２个碱基不相同,在１７５位密码子（５２３～５２５位的碱基）分别是ＣＧＧ和ＣＡＡ,即精氨酸（Ｒ）的位置上,中国人是谷胺酰氨（Ｇ）．而与测得的韩国人序列比较,在相应位置的氨基酸是相同的     

Preparation of insoluble fibroin films and its tensile property

Silk fibroin is becoming a promising biomate-rial because of its excellent biocompatibility. However, theregenerated fibroin is usually soluble in water and its me-chanical properties should be improved. Although manymethods, such as adding other polymers or treating withmethanol, can ameliorate the mechanical properties andinsolubility, the biocompatibility of fibroin is usually dam-aged in these processes. In this article, it is first reported thatthe insoluble fibroin films are directly prepared withoutmethanol treatment. According to the results of Fouriertransform infrared spectroscope (FTIR) and the X-ray dif-fraction (XRD), the amount of IS-sheet conformation in-creased with the increasing of concentration. When fibroinfilms are dried from 15 wt% at 60℃, the films become in-soluble in water. More importantly, The tensile strength andelongation of the insoluble fibroin films dried from 15% so-lution at 60℃ reached 15.9 MPa and 49.4% respectively in the wet state, which is distinctly superior to the fibroin films treated with methanol.     

Characterization of codon usage bias in the dUTPase gene of duck enteritis virus

A comparative analysis of the codon usage bias in the newly discovered dUTPase gene （Assigned Accession No.： DQ486149） of the duck enteritis virus （DEV） and the dUTPase gene of 32 reference herpesviruses was performed. The results indicated that the DEV dUTPase gene encodes a protein of 477 amino acids, which includes five conserved motifs with a 3-1-2-4-5 arrangement. The codon adaptation index （CAI）, effective number of codons （ENC）, and GC3s values indicated synonymous codon usage bias in the dUTPase gene of herpesviruses, and this synonymous bias was correlated with host evolution. The codon usage patterns of the DEV dUTPase gene were phylogenetically conserved and similar to that of the dUTPase genes of the avian alphaherpesvirus. Although codon usage in each microorganism was different, there were no strain-specific differences among them. Sixty-one codons in the predicted polypeptide, with a strong bias towards A and T at the third codon position, were used. Comparison of the codon usage in the dUTPase gene of different organisms revealed that there were 19 codons showing distinct codon usage differences between the DEV and Escherichia coli dUTPase genes; 16 between the DEV and yeast dUTPase genes; and 15 between the DEV and human dUTPase genes. Analysis of variance （ANOVA） showed significant differences between the DEV and yeast dUTPase genes （r = 0.536, P 〈 0.01）. The extent of codon usage bias in the DEV dUTPase gene was highly correlated with the gene expression level, therefore the results may provide useful information for gene classification and functional studies.     

高含量再生丝素蛋白水溶液的热性能研究

用差示扫描量热仪(DSC),对不同含量的再生丝素蛋白水溶液(质量分数为12%~42%),以及同一含量不同剪切速率(1 000 s-1和2 000 s-1)下溶液热性能的变化进行了对比研究。结果表明,随着含量的增大,再生丝素蛋白水溶液发生相转变的温度逐渐降低;同样,经剪切后的溶液发生相转变的温度均向低温漂移,且随着剪切作用的加强,向低温的漂移越明显。试验表明,丝素水溶液含量的增加和剪切作用可加速溶液从无规线团或α-螺旋向β-折叠构象的转变速度,且随着剪切速率的提高,构象转变的速度也加快。     

Complete sequence and gene organization of the mitochondrial genome of Batocera lineolata Chevrolat (Coleoptera: Cerambycidae)

Batocera lineolata Chevrolat,1852 is an important forest pest that is found mainly in China,Vietnam,India,and Japan.The complete mitochondrial genome(mitogenome) sequence of B.lineolata was determined by long polymerase chain reaction(PCR) and conserved primer walking approaches.The results showed that the entire mitogenome is 15418 bp long with 74.48% A+T content.The positions and arrangement of the 37 genes encoded by the genome are identical to the mitogenomes of two other longhorn beetles for which the complete gene content and arrangement are publicly available.All protein-coding genes start with the ATN codon that is a typical initiation codon in insects.All transfer RNAs(tRNAs) were predicted to form the standard clover-leaf structure,except for tRNA Ser(AGN),which lacks the dihydrouridine(DHU) arm.The most unusual feature that was found was the use of TCT as the tRNA Ser(AGN) anticodon instead of the GCT that is used in most other arthropods.The lack of tandem repeat motif in the 735 bp long A+T-rich region was another unusual feature of the B.lineolata mitogenome.The short,highly conserved polythymidine stretch that was previously described in the Orthoptera and Diptera orders was also present in the A+T-rich region of the B.lineolata(order Coleoptera) mitogenome.The sequence and annotation of the mitogenome of B.lineolata will provide further insights into the diversity and evolution of the Cerambycidae family of long-horned beetles.The B.lineolata mitogenome sequence has been deposited in GenBank(http://www.ncbi.nlm.nih.gov/Genbank/) under accession number JN986793.     

Adaptive evolution and structure modeling of rbcL gene in Ephedra

The adaptive evolution of the rbcL gene in 14 species of Ephedra and 9 other gymnosperms was tested by the PAML program. Three large-scale selections were found in the catalytic region of the RbcL protein by branch model. Species in Ephedra with positive selection at the 365th codon site in rbcL gene were detected by branch-site model. 3D structure modeling showed that the 365th codon could change the spatial structure of the β-sheet (βG, βH, or βE) of the RbcL protein. These changes may be associated with ...     

几何计算及其理论研究

提出了一种新的几何计算理论.在几何基础层，充分利用笛卡儿创立的坐标几何思想，用几何代数化方法构建二、三维基本的几何代数基（简称几何基)，可利用它的序列建立高一层次的几何基.在几何处理层，用几何方法解决几何问题，寻求几何问题的几何基求解序列.对几何引入方向性，统一几何的表示，简化几何基序列的求解过程.并从理论上探索解决几何奇异问题的完整解决方案，形成一个统一、规范的几何计算体系.由此实现莱布尼茨式的通过几何语言直接处理几何体的宏伟设想.