青海沙棘果粉抗氧化活性的研究

为了深入了解青海沙棘果粉的品质,助其实现规模化生产,从1,1-二苯基苦基苯肼自由基(DPPH·)体系、羟基自由基(·OH)体系、超氧阴离子自由基(O-2·)体系和亚硝酸盐(NO-2)的清除效果方面着手,研究了其体外清除自由基活性。结果表明,青海沙棘果粉对DPPH·、·OH、O-2·和NO-2均有清除作用,且对DPPH·、·OH的清除能力优于O-2·和NO-2。     

大鼠皂素蜕膜心肌肌钙蛋白Ca^2＋敏感性的实验研究

采用５００μｇ／ｍｌ皂素溶液浸浴大鼠右室乳头肌３０ｍｉｎ制备蜕膜心肌纤维标本。经撤除ＭｇＡＴＰ法检验,肌膜通透性明显增高。用含不同浓度Ｃａ￣(２＋)（以Ｃａ￣(２＋)浓度负对数ｐＣａ表示）的激活液进行顺序激活,记录Ｃａ￣(２＋)激活张力,其张力－ｐＣａ关系曲线描述了肌钙蛋白（ＴＮ）对Ｃａ￣(２＋)的亲和特性;曲线中位值ｐＣａ＿(５０)（即产生５０％最大张力所对应的ｐＣａ）是反映ＴＮＣａ￣(２＋)敏感性的定量指标。本实验观察到大鼠右室乳头肌张力－ｐＣａ关系曲线呈Ｓ形,测得ｐＣａ＿(５０)为５．７２７±０．０８６（ｎ＝１６）;撤除激活液中磷酸肌酸和磷酸肌酸激酶,张力－ｐＣａ曲线左移位,ｐＣａ＿(５０)增高至６．１９４±０．１２１（Ｐ＜０．０１）。     

白粉藤的木脂素和三萜成分

从白粉藤（Cissus repens Lank）地上部分分离得到5个木脂素和8个三萜,其中一个木脂素是新化合物,它的结构通过波谱分析和碱水解的方法鉴定为：（＋）-异落叶松树脂醇-9′-（2-对-香豆酰）-O-β-D-吡喃木糖苷（1）。其余化合物分别是：（＋）-异落叶松树脂醇-9′-O-β-D-吡喃木糖苷（2）,（＋）-Lyoniside（3）,（—）-开环异落叶松树脂醇-9-O-β-D-吡喃木糖苷（4）,（7′R,8′S）-4′-hydroxy-3′,5-dimethoxy-7,8′-dihydrobenzofuran-1-propanolneolignan-9′-O-β-D-xylopyranoside（5）,木栓酮（6）,表木栓醇（7）,蒲公英赛醇乙酸酯（8）,熊果酸（9）,2α-羟基乌索酸（10）,积雪草酸（11）,Niga-ichigoside F1（12）,羽扇豆醇（13）。这些化合物都是首次从该植物中分离得到。     

Polymorphisms of the DNA Methyltransferase 1 Associated with Reduced Risks of Helicobacter pylori Infection and Increased Risks of Gastric Atrophy

Introduction

DNA methyltransferase-1(DNMT1) is an important enzyme in determining genomic methylation patterns in mammalian cells. We investigated the associations between SNPs in the DNMT1 gene and risks of developing H. pylori seropositivity, gastric atrophy and gastric cancer in the Chinese population.

Methods

The study consisted of 447 patients with gastric cancer; 111 patients with gastric atrophy; and 961 healthy controls. Five SNPs, rs10420321, rs16999593, rs8101866, rs8111085 and rs2288349 of the DNMT1 gene were genotyped. Anti-H.pylori IgG was detected by ELISA. Gastric atrophy was screened by the level of serum pepsinogen Ιand II and then confirmed by endoscopy and histopatholgical examinations.

Results

The age- and sex-adjusted OR of H. pylori seropositivity was 0.67 (95%CI: 0.51–0.87) for rs8111085 TC/CC genotypes, significantly lower than the TT genotype in healthy controls. The adjusted OR of H.pylori seropositivity was 0.68 (95%CI: 0.52–0.89) for rs10420321 AG/GG genotypes. In addition, patients carrying rs2228349 AA genotype have a significantly increased risk for H.pylori seropositivity (OR = 1.67; 95%CI: 1.02–2.75). Further haplotype analyses also showed that the ATTTG and ATCTA are significantly associated with increased risks in H.pylori infection compared to the GTCCG haplotype (OR = 1.38, 95%CI: 1.08–1.77; OR = 1.40, 95% CI: 1.09–1.80). The adjusted ORs of gastric atrophy were 1.66 (95%CI: 1.06–2.61) for rs10420321 GG genotype, and 1.67 (95%CI 1.06–2.63, P = 0.03) for rs8111085 CC genotype, but no association was found between SNPs in the DNMT1 gene and risk of developing gastric cancer.

Conclusions

Individuals with rs10420321 GG and rs8111085 CC genotype of the DNMT1 gene were associated with reduced risks for H.pylori infection. On the other hand, higher risks of gastric atrophy were found in the carriers with these two genotypes compared to other genotypes. Our results suggested that SNPs of DNMT1 could be used as genotypic markers for predicting genetic susceptibilities to H.pylori infection and risks in gastric atrophy.     

葡酒红菇的生态学研

本研究通过４年对葡酒红菇生态学的调查观察,掌握葡酒红菇的生态环境,以便人工模拟生态环境,栽培引种葡酒红菇。研究结果表明：葡酒红菇是一种外生菌根菌,在相对稳定的生态坏境条件下,平均温度２５—２７℃,相对湿度８０％－１００％,降雨量（土壤含水量）达４０％以上,均有红菇发生,而降雨量力其主导生态因子。每年在夏秋高温闷热阵雨之后为发生盛期。发生地以山中下部为多,林间郁团度０．８左右,土壤ｐＨ值５．６—７．０,其共生树种以壳斗科为主。     

伤胁迫对蚕豆叶片中茉莉酸分布的影响

在植物应对伤害等环境刺激的反应中,已知茉莉酸(JA)作为一种重要的信号分子在植物体内长距离运输,但目前对JA的细胞和亚细胞定位知之甚少。本研究用免疫荧光显微镜技术和免疫胶体金电镜技术证明茉莉酸分布在蚕豆叶片叶肉细胞的叶绿体、表皮细胞的细胞壁、保卫细胞的细胞壁、细胞质、叶绿体和细胞核上。其中保卫细胞的叶绿体和细胞核是JA分布的主要场所。叶片的局部灼伤可提高JA在质外体和气孔保卫细胞中的水平。由此推测,伤胁迫下JA分配的改变可能与植物体防御反应密切相关,并参与了对气孔运动的调控。     

肾性高血压大鼠左室等容峰压一容积关系及其对异丙肾上腺素的反应

本研究观察了肾血管性高血压大鼠(RHR)肥大左室的等容峰压-容积关系(PIPVR)和左室压力最大上升速率-容积关系(PRPVR)的曲线特征,并用两曲线的升支斜率E_(max)和dE/dt_(max)及曲线的特征参数评价了RHR(8周)左室的收缩能力及其对异丙肾上腺素的反应性。结果显示,大鼠高血压8周后左室发生明显肥大;与同龄假手术对照大鼠(SOR)相似,RHR的PIPVR和PRPVR也呈指数曲线形式;与SOR比较,RHR左室的PIPVR和PRPVR左上移位,E_(max)、dE/dt_(max)显著增大(P均<0.01),分别增加78％和97％,曲线的特征参数K_1、K_2、B_1和B_2也明显高于SOR(P均<0.05),分别增加37％、32％、25％和57％;灌注异丙肾上腺素(0.94μg/min)后,上述指标的增加幅度均较SOR低(P均<0.05)。结果提示,压力超负荷心肌肥大并不影响PIPVR的非线性特征,基础状态下,RHR肥大左室的收缩能力增强,但对异丙肾上腺素的反应性降低。     

A Na+/H+ antiporter gene of the moderately halophilic bacterium Halobacillus dabanensis D-8T: cloning and molecular characterization

A gene encoding a Na(+)/H(+) antiporter was cloned from a chromosomal DNA of Halobacillus dabanensis strain D-8(T) by functional complementation. Its presence enabled the antiporter-deficient Escherichia coli strain KNabc to survive in the presence of 0.2 M NaCl or 5 mM LiCl. The gene was sequenced and designated as nhaH. The deduced amino-acid sequence of NhaH consists of 403 residues with a calculated molecular mass of 43,481 Da, which was 54% identical and 76% similar to the NhaG Na(+)/H(+) antiporter of Bacillus subtilis. The hydropathy profile was characteristic of a membrane protein with 12 putative transmembrane domains. Everted membrane vesicles prepared from E. coli cells carrying nhaH exhibited Na(+)/H(+) as well as Li(+)/H(+) antiporter activity, which was pH-dependent with highest activities at pH 8.5-9.0 and at pH 8.5, respectively. Moreover, nhaH confers upon E. coli KNabc cells the ability to grow under alkaline conditions.     

Up-regulation of the fidelity of human DNA polymerase lambda by its non-enzymatic proline-rich domain

DNA repair pathways are essential for maintaining genome stability. DNA polymerase beta plays a critical role in base-excision repair in vivo. DNA polymerase lambda, a recently identified X-family homolog of DNA polymerase beta, is hypothesized to be a second polymerase involved in base-excision repair. The full-length DNA polymerase lambda is comprised of three domains: a C-terminal DNA polymerase beta-like domain, an N-terminal BRCA1 C-terminal domain, and a previously uncharacterized proline-rich domain. Strikingly, pre-steady-state kinetic analyses reveal that, although human DNA polymerase lambda has almost identical fidelity to human DNA polymerase beta, the C-terminal DNA polymerase beta-like domain alone displays a dramatic, up to 100-fold loss in fidelity. We further demonstrate that the non-enzymatic proline-rich domain confers the increase in fidelity of DNA polymerase lambda by significantly lowering incorporation rate constants of incorrect nucleotides. Our studies illustrate a novel mechanism, in which the DNA polymerase fidelity is controlled not by an accessory protein or a proofreading exonuclease domain but by an internal regulatory domain.     

DNA lesion alters global conformational dynamics of Y-family DNA polymerase during catalysis

A major product of oxidative damage to DNA, 8-oxo-7,8-dihydro-2'-deoxyguanine (8-oxoG), can lead to genomic mutations if it is bypassed unfaithfully by DNA polymerases in vivo. However, our pre-steady-state kinetic studies show that DNA polymerase IV (Dpo4), a prototype Y-family enzyme from Sulfolobus solfataricus, can bypass 8-oxoG both efficiently and faithfully. For the first time, our stopped-flow FRET studies revealed that a DNA polymerase altered its synchronized global conformational dynamics in response to a DNA lesion. Relative to nucleotide incorporation into undamaged DNA, three of the four domains of Dpo4 undertook different conformational transitions during 8-oxoG bypass and the subsequent extension step. Moreover, the rapid translocation of Dpo4 along DNA induced by nucleotide binding was significantly hindered by the interactions between the embedded 8-oxoG and Dpo4 during the extension step. These results unprecedentedly demonstrate that a Y-family DNA polymerase employs different global conformational dynamics when replicating undamaged and damaged DNA.