DR-70~(TM)和CA50肺癌免疫测定的应用价值

目的 :评价DR 70 TM和CA5 0两种肿瘤标志物在肺癌检测中的价值。方法 :应用DR 70 TMELISA和CA5 0IRMA对77例肺癌患者 ,42例健康者及 48例其它肺癌者进行对照检测。结果 :各组DR 70 TMCA5 0测定值比例显示肺癌组平均值显著其它两组 (P <0 .0 1)。DR 70 TM检测的敏感性和有效性高于CA5 0检测 (83.1%比 6 6 .2 % ,85 .7%比 75 .6 % )。两项联合检测的敏感性 ,有效性分别为 96 .1% ,93.1% ,优于单项DR 70 TM 检测的 83.1% ,85 .7% ,及CA5 0的 6 6 .2 9%、75 .6 %。结论 :DR 70 TM在肺癌检测中有较大应用价值。DR 70 TM 和CA5 0两项联合检测是较理想的肺癌检测组合     

Association of some potential hormone response elements in human genes with the Alu family repeats.

Short interspersed repeats of the Alu family located in promoters of some human genes contain high-affinity binding sites for thyroid hormone receptor, retinoic acid receptor and estrogen receptor. The standard binding sites for the receptors represent variants of duplicated AGGTCA motif with different spacing and orientation (direct, DR, or inverted, IR), and Alu sequences were found to have functional DR-4, DR-2 or variant IR-3/IR-17 elements. In this study we analyzed distribution and abundance of the elements in a set of human genomic sequences from GenBank and their association with Alu repeats. Our results indicate that a major fraction of potentially active DR-4, DR-2 and variant IR-3/IR-17 elements in the genes is located within Alu repeats. Alu-associated DR-2 elements are conserved in primate evolution. However, very few Alu have potential DR-3 glucocorticoid-response elements. Gel-shift experiments with the probe (AUB) corresponding to the consensus Alu sequence just upstream of the RNA polymerase III promoter B-box and containing duplicated AGGTCA motif indicate that the probe interacts in a sequence-specific manner with human nuclear proteins which bind to standard IR-0, DR-1, DR-4 or DR-5 elements. The AUB sequence was also able to promote thyroid hormone-dependent trans-activation of a reporter gene. The results support the view that Alu retroposons played an important role in evolution of regulation of the primate gene expression by nuclear hormone receptors.     

Multiple Resistance and Biochemical Mechanisms of Dicofol Resistance in Tetranychus urticae (Acari: Tetranychidae)

A field colony of Tetranychus urticae (Koch) resistant to dicofol was selected with dicofol successively for 20 generations to produce the DR-20 strain. Resistance and multiple resistance levels of the DR-20 strain to 15 acaricides were determined using a spray bioassay. The DR-20 strain was extremely resistant to dicofol [resistance ratio (RR), 465]. The strain showed extremely strong resistance to acrinathrin (RR, 373) and benzoximate (RR, 197) and strong resistance to bromopropylate (RR, 136), fenbutatin oxide (RR, 65), fenpropathrin (RR, 70), fenpyroximate (RR, 68), and pyridaben (RR, 63). A RR of 11–29 was observed with abamectin, fenazaquin, milbemectin, propagite, and tebufenpyrad. The DR-20 strain exhibited low levels of resistance (RR<3) to azocyclotin and chlorfenapyr. In comparative assays with detoxifying enzymes, the DR-20 strain showed 4.7-fold higher activity in p-nitroanisole-O-demethylation and 1.6-fold higher activities in both α- and β-naphthyl acetate hydrolysis. Synergist experiments with different metabolic inhibitors revealed that piperonyl butoxide, iprobenfos, triphenyl phosphate, and 4, 4-dichloro-α-methyl benzhydrol had little or no synergistic activity in the susceptible and DR-20 strains. These results suggest that employment of certain acaricides with little multiple resistance will be useful for the management of dicofol resistance in the field.     

Aerobic biodegradation of Azo dye by Bacillus cohnii MTCC 3616; an obligately alkaliphilic bacterium and toxicity evaluation of metabolites by different bioassay systems

An obligate alkaliphilic bacterium Bacillus cohnii MTCC 3616 aerobically decolorized a textile azo dye Direct Red-22 (5,000 mg?l^?1) with 95 % efficiency at 37 °C and pH?9 in 4 h under static conditions. The decolorization of Direct Red-22 (DR-22) was possible through a broad pH (7–11), temperature (10–45 °C), salinity (1–7 %), and dye concentration (5–10 g?l^?1) range. Decolorization of dye was assessed by UV–vis spectrophotometer with reduction of peak intensity at 549 nm (λ _max). Biodegradation of dye was analyzed by Fourier transform infrared spectroscopy (FTIR) and high-performance liquid chromatography (HPLC). The FTIR spectrum revealed that B. cohnii specifically targeted azo bond (N=N) at 1,614.42 cm^?1 to break down Direct Red-22. Formation of metabolites with different retention times in HPLC analysis further confirmed the degradation of dye. The phytotoxicity test with 5,000 mg?l^?1 of untreated dye showed 80 % germination inhibition in Vigna mungo, 70 % in Sorghum bicolor and 80 % in Vigna radiata. No germination inhibition was noticed in all three plants by DR-22 metabolites at 5,000 mg?l^?1. Biotoxicity test with Artemia salina proved the lethality of the azo dye at LC₅₀ of 4 and 8 % for degraded metabolites by causing death of its nauplii compared to its less toxic-degraded metabolites. Bioaccumulation of dye was observed in the mid-gut of A. salina. The cytogenotoxicity assay on the meristematic root tip cells of Allium cepa further confirmed the cytotoxic nature of azo dye (DR-22) with decrease in mitotic index (0.5 % at 500 ppm) and increase in aberrant index (4.56 %) over 4-h exposure period. Genotoxic damages (lagging chromosome, metaphase cluster, chromosome bridges, and dye accumulation in cytoplasm) were noticed at different stages of cell cycle. The degraded metabolites had negligible cytotoxic and genotoxic effects.     

ACC氧化酶和ACC合成酶反义RNA融合基因导入番茄和乙烯合成的抑制

从番茄品种强力米寿的总DNA中克隆番茄果实特异启动子2A11,以番茄成熟果实的RNA为模板,进行RT-PCR扩增,克隆番茄全长的ACC氧化酶基因和ACC合成酶基因片段。完成两个基因的克隆和测序后,将888bp的番茄ACC氧化酶基因和943bp的ACC合成酶基因片段串联,构成全长1837bp的融合基因。将该融合基因以反义的方向插入植物双元载体pYPX145中番茄果实表达特异启动子下游,获得ACC氧化酶基因和ACC合成酶基因融合的植物双元载体pOSACC。该载体外源基因表达单元的两端含两个烟草SAR序列,利于转基因的稳定遗传。以番茄栽培品种合作903子叶和下胚轴为外植体,利用根癌农杆菌进行基因转化,通过200mg／L卡那霉素选择和GUS检测,获得了105株番茄GUS阳性植株,转基因番茄果实在当代表现明显耐贮特点。经过4代的耐贮和果实农艺性状的综合选择,获得了两个表现良好的株系DR-1和DR-2,两株系果实乙烯释放量显著下降,是未转基因材料的9．5％,番茄的贮存期在50天以上。     

Isolation and identification of peptides from simulated gastrointestinal digestion of preserved egg white and their anti-inflammatory activity in TNF-α-induced Caco-2 cells

Simulated gastrointestinal digestion of preserved egg white (SGD-PEW) exerts anti-inflammatory effects on Caco-2 cells and a mouse model of DSS-induced colitis. Here, we aimed to separate peptides derived from SGD-PEW and evaluate their anti-inflammatory effects using an in vitro inflammatory model. Six peptides were isolated and identified. DEDTQAMPFR (DR-10), DEDTQAMPF (DF-9), MLGATSL (ML-7) and MSYSAGF (MF-7) significantly inhibited IL-8 secretion and markedly decreased gene expression, including TNF-α, IL-8, IL-6, IL-1β and IL-12 and promoted IL-10 gene expression in Caco-2 cells. DR-10, DF-9, ML-7 and MF-7 significantly inhibited the phosphorylation of JNK. In the meantime, DR-10 and DF-9 significantly reduced the phosphorylation of IκB and p38. These results indicated that ML-7 and MF-7 exerted their anti-inflammatory activity through the MAPK signaling pathway in TNF-α-induced Caco-2 cells. Whereas, DR-10 and DF-9 inhibited the NF-κB and MAPK signaling pathways. The results suggested that DR-10, DF-9, ML-7 and MF-7 derived from SGD-PEW may be a new type of prophylactic food for the treatment of inflammation.     

Morphine-like analgesic actions of enkephalin-like peptides containing the Tyr-Arg unit

The analgesic actions of some synthetically prepared peptides having the Tyr-D-Arg unit at the N terminal portion of met- and leu-enkephalin were measured by the intra-cisternal injection method in mice. Among them, Tyr-D-Arg-Gly-Phe (DR-4) induced the most potent naloxone-reversible analgesia and was also effective by s.c. injection. DR-4 showed the good affinity to mu-receptor, and the resistance to the enzymatic degradation.     

Only one DQ-beta restriction fragment pattern of each DR specificity is associated with insulin-dependent diabetes

Insulin-dependent diabetes is generally associated with the serologic HLA-DR specificities 3 and 4, in particular with DR-3,4 heterozygosity. The disease is negatively associated with DR-2. To investigate these associations further at the genomic level, DNA from 13 families with a proband having insulin-dependent diabetes, from 11 other individuals with the same disease, and from HLA-DR-matched control individuals was subjected to restriction fragment analysis. Three different enzymes (Bam HI, Eco RI, and Pvu II) and cDNA clones for three HLA-D region class II antigen alpha- and beta-chains (DR-beta, DQ-beta, and DQ-alpha) were used. In six families, a total of 11 siblings HLA-DR-identical to the proband were examined. There was no discrepancy between the hybridization patterns of the proband and those of the DR-identical siblings. Two different DQ-B fragment patterns were detected with each one of the serologic specificities DR-2 and DR-4. In both cases, only one of the patterns correlated significantly with diabetes. Thus, DQ-beta genomic hybridization may be used in conjunction with HLA-DR typing to identify individuals with higher relative risk to acquire insulin-dependent diabetes. These results may suggest that insulin-dependent diabetes is associated with the DQ rather than with the DR locus.     

The structure and function of the replication initiator protein (Rep) of pSC101: an analysis based on a novel positive-selection system for the replication-deficient mutants

Plasmid pSC101 encodes a 37.5 kDa Rep (RepA) protein, which binds to three 21-base repeats (DR-1, DR-2, and DR-3) in the replication origin region (ori) of the plasmid to initiate replication. Rep also binds to two palindromic sequences (IR-1 and IR-2) which overlap the rep promoter. The binding of Rep to IR-2 represses the production of Rep itself. It is highly likely that the balance of these functions of Rep plays a major role in controlling the copy number of pSC101. In this study, we developed a positive-selection system for replication-deficient mutants of the initiator protein. This system can be applied to the study of other replication systems by changing ori and rep of pSC101 to the corresponding genes. Thirty-four replication-deficient (Ini(-)) mutants were isolated with this system, and analyzed as to the relation between the structure and function of the Rep protein. Seventeen of these 34 Ini(-) mutants were found to lack auto-repressor activity as well as initiator activity. DNA sequence analysis showed that one-third (from the C-terminus) of Rep is dispensable for the auto-repressor activity, while the initiator activity seems to require the whole protein.     

Fragment based search for small molecule inhibitors of HIV-1 Tat-TAR

Basic molecular building blocks such as benzene rings, amidines, guanidines, and amino groups have been combined in a systematic way to generate ligand candidates for HIV-1 TAR RNA. Ranking of the resulting compounds was achieved in a fluorimetric Tat-TAR competition assay. Although simple molecules such as phenylguanidine are inactive, few iteration steps led to a set of ligands with IC₅₀ values ranging from 40 to 150 μM. 1,7-Diaminoisoquinoline 17 and 2,4,6-triaminoquinazoline 22 have been further characterized by NMR titrations with TAR RNA. Compound 22 is bound to TAR at two high affinity sites and shows slow exchange between the free ligand and the RNA complex. These results encourage investigations of dimeric ligands built from two copies of compound 22 or related heterocycles.     

ACC氧化酶和ACC合成酶反义RNA融合基因导入番茄和乙烯合成的抑制

从番茄品种强力米寿的总DNA中克隆番茄果实特异启动子2A11,以番茄成熟果实的RNA为模板,进行RT-PCR扩增,克隆番茄全长的ACC氧化酶基因和ACC合成酶基因片段。完成两个基因的克隆和测序后,将888bp的番茄ACC氧化酶基因和943bp的ACC合成酶基因片段串联,构成全长1837bp的融合基因。将该融合基因以反义的方向插入植物双元载体pYPX145中番茄果实表达特异启动子下游,获得ACC氧化酶基因和ACC合成酶基因融合的植物双元载体pOSACC。该载体外源基因表达单元的两端含两个烟草SAR序列,利于转基因的稳定遗传。以番茄栽培品种合作903子叶和下胚轴为外植体,利用根癌农杆菌进行基因转化,通过200mg/L卡那霉素选择和GUS检测,获得了105株番茄GUS阳性植株,转基因番茄果实在当代表现明显耐贮特点。经过4代的耐贮和果实农艺性状的综合选择,获得了两个表现良好的株系DR-1和DR-2,两株系果实乙烯释放量显著下降,是未转基因材料的9.5%,番茄的贮存期在50天以上。     

一株青藏高原冻土溶栓菌的筛选及鉴定

【目的】心脑血管疾病是一种世界性疾病,严重危害人类健康,溶栓酶是治疗该病的有效药物之一。而极端环境中的溶栓微生物因其特殊的生存方式,可能分泌高效、安全的新型溶栓酶。因此,为了获得这种具有特殊功能的溶栓酶,我们从青藏高原高海拔冻土中进行了溶栓菌的筛选。【方法】首先,本文通过血粉-琼脂平板初步筛选具有血粉水解功能的菌株,然后对其进行体外溶栓试验以检验其人工血栓溶解功能,并用纤维蛋白平板法测定其纤溶活性,最后通过生理生化试验和16S rRNA基因序列分析方法对该菌进行分类鉴定。【结果】本文从青海省玉树藏族自治州海拔4300 m的冻土样品中筛选获得了菌株DR-536,不仅具有水解血粉的功能,还具有体外溶栓功能,且能够水解纤维蛋白,纤溶活性为51.80 IU/mL(以尿激酶为标准)。最后,分类鉴定结果显示菌株DR-536是一株金黄节杆菌(Arthrobacter aurescens)。【结论】本文首次从青藏高原高海拔土壤中进行了溶栓菌的筛选,并获得了一株新型溶栓菌,为进一步研究和开发高效、安全的新型溶栓酶提供了菌源。