A study on the localization and distribution of GnRH and its receptor in rat submaxillary glands by immunohistochemical,in situ hybridization and RT-PCR

We investigated the rat submaxillary gland for the presence of GnRH and GnRH receptors, the localization and colocalization of GnRH, GnRH receptor and their mRNA, and studied the sequence of GnRH receptor complementary DNA (cDNA) by immunohistochemistry, in situ hybridization and RT-PCR. The results showed that GnRH and GnRH receptor immunoreactive materials were colocalized in the epithelial cells of the serous acinus and glandular duct. The GnRH and GnRH receptor mRNA hybridization signals were detected in the above cells. The sequence obtained from the RT-PCR product was identical to the published cDNA sequence of GnRH receptor in the rat pituitary. The results suggested that the rat submaxillary gland was capable of synthesizing GnRH and GnRH receptors. GnRH may be involved in the functional regulation of the submaxillary gland through autocrine or paracrine activity.     

Measuring dynamics of caspase-8 activation in a single living HeLa cell during TNFalpha-induced apoptosis

In this study, we reported the first measurement of the dynamics of activation of caspase-8 in a single living cell. This measurement was conducted using a specially developed molecular sensor based on the FRET (fluorescence resonance energy transfer) technique. This sensor was constructed by fusing a CFP (cyan fluorescent protein) and a YFP (yellow fluorescent protein) with a linker containing a tandem caspase-8-specific cleavage site. The change of the FRET ratio upon cleavage was larger than 4-fold. Using this sensor, we found that during TNFalpha-induced apoptosis, the activation of caspase-8 was a slower process than that of caspase-3, and it was initiated much earlier than the caspase-3 activation. Inhibition of caspase-9 delayed the full activation of caspase-3 but did not affect the dynamics of caspase-8. Results of these single-cell measurements suggested that caspase-3 was activated by caspase-8 through two parallel pathways during TNFalpha-induced apoptosis in HeLa cells.     

A single point mutation resulting in an adversely reduced expression of DPM2 in the Lec15.1 cells

Mammalian dolichol-phosphate-mannose (DPM) synthase consists of three subunits, DPM1, DPM2, and DPM3. Lec15.1 Chinese hamster ovary cells are deficient in DPM synthase activity. The present paper reports that DPM1 cDNA from wild type and Lec15.1 CHO cells were found to be identical, and transfection with CHO DPM1 cDNA did not reverse the Lec15.1 phenotype. Neither did a chimeric cDNA containing the complete hamster DPM1 open reading frame fused to the Saccharomyces cerevisiae DPM1 C-terminal transmembrane domain. In contrast, Lec15.1 cells were found to have a single point mutation G29A within the coding region of the DPM2 gene, resulting in a glycine to glutamic acid change in amino acid residue 10 of the peptide. Moreover, mutant DPM2 cDNA expressed a drastically reduced amount of DPM2 protein and poorly corrects the Lec15.1 cell phenotype when compared with wild type CHO DPM2 cDNA (G(29) form).     

莫能菌素高产菌株的诱变与筛选

采用紫外线对现有生产菌株进行诱变处理,再运用筛选剂丙酸、丁酸等对其进行选育,得到高产菌株M-3-01。投入中试车间发酵罐中,发酵效价达到50560×103u.L-1,其发酵能力比出发菌株提高了26%。     

施氮时期对玉米土壤硝态氮含量变化及氮盈亏的影响

在“郑单 95 8”(9株 / m2 )组成的土 -植系统 ,研究了不施氮、基施氮 10叶展追氮、基施氮 吐丝期追氮和基施氮 乳熟期追氮共 4个处理下 0～ 2 0 0 cm的土壤 NO- 3- N含量在夏玉米生长期间的变化和土壤氮素的表观盈亏量 ,结果表明 :2 0 cm以上的土壤 NO- 3- N含量以大口期为界、2 0 cm以下的土壤 NO- 3- N含量以吐丝期为界前降后升。在 0～ 2 0 cm土层 ,与不施氮相比 ,施氮能增加土壤 NO- 3- N含量 ,而且吐丝期和乳熟至成熟阶段的 NO- 3- N含量在 10叶展期和吐丝期各自追氮后均显著增加。在 2 0～4 0 cm土层 ,乳熟期的 NO- 3- N含量施氮后明显比不施氮高。在 80 cm以下土层 ,施氮后的土壤 NO- 3- N含量明显比不施氮高 ;追氮期相比 ,后一追氮处理在乳熟期和成熟期的 NO- 3- N含量均比前一追氮处理明显增加 ,其中成熟期基施氮 乳熟期追氮处理在 16 0～ 2 0 0 cm土层的 NO- 3- N含量比基施氮 吐丝期追氮处理 (为 2 5 .3m g N/ kg(干土 ) )高 16 %。土壤氮素的表观盈余发生在吐丝期之前且 80 %以上盈余量出现在大口期前 ,表观亏损出现在吐丝期以后且其亏损量在乳熟期前后各占一半。经玉米季后 ,本试验中不施氮处理出现表观盈余 (为 5 6 .3kg N/ hm2 ) ;施氮后表观盈余量增加 ,主要是施氮减少了吐丝以后     

(2-amino-phenyl)-amides of omega-substituted alkanoic acids as new histone deacetylase inhibitors

A variety of omega-substituted alkanoic acid (2-amino-phenyl)-amides were designed and synthesized. These compounds were shown to inhibit recombinant human histone deacetylases (HDACs) with IC(50) values in the low micromolar range and induce hyperacetylation of histones in whole cells. They induced expression of p21WAF1/Cip1 and caused cell-cycle arrest in human cancer cells. Compounds in this class showed efficacy in human tumor xenograft models.     

Methylation profiling of urothelial carcinoma in bladder biopsy and urine

OBJECTIVE: To test DNA methylation profiling in detection of urothelial carcinoma in urine. STUDY DESIGN: Thirty-three bladder specimens were analyzed for the DNA p16INK4a, RASSF1, APC, GSTP, E-Cad and CyclinD2 genes to determine if there is a difference in gene methylation between benign and malignant cases. Urine samples were analyzed in a feasibility study. Finally, methylation profiles of urine samples were obtained and compared with follow-up biopsy diagnoses. RESULTS: We found methylated genes in 18% benign, 37% urothelial carcinoma in situ and 93% infiltrating urothelial carcinoma cases (p = 0.001). Methylation profiles from the 18 urine samples revealed a significantly higher prevalence of methylated genes in carcinoma cases than benign cases (100% vs. 50%, p = 0.025). We analyzed methylation profiles in 37 cytologically atypical urine samples with malignant or benign diagnosis on surgical follow-up andfound that only APC (55% in malignant vs. 0% in benign, p=0.025) and CyclinD2 were differentially methylated (35% in malignant vs. 0% in benign, p=0.2) while p14ARF, p16INK4a, RASSF1, GSTP and E-Cad had similar methylation profiles. CONCLUSION: These results suggest that methylation of p14ARF, p16INK4a, RASSF1, GSTP and E-Cad genes may not accurately identify carcinoma, but methylated APC and CyclinD2 might be useful biomarkers for urothelial carcinoma in urine.     

元宝草化学成分的研究

目的　研究中药元宝草 (HypericumsampsoniiH .)的化学成分。 方法　用乙醇或甲醇回流提取 ,聚酰胺柱色谱和重结晶分离 ,FT -IR ,MS ,UV ,1HNMR等方法确定结构。结果　分得了 4个化学成分 ,分别为芦丁 ,金丝桃苷 ,槲皮素和金丝桃素。结论　 4种化合物均为首次从元宝草中分得和检出。     

金铁锁的一个新三萜成分

金铁锁(Psammosilene tunicoides W.C.Wu et C.Y.Wu)系石竹科金铁锁属单一种植物,产于我国西南部。该植物为传统中药,具止血、止痛、活血等多种功效。其根部富含皂甙成分,有镇痛、抗炎和抑菌等生理活性。我们曾报道了将其根中提取的粗皂甙酸性水解后,分离到的六个齐墩果酸型三萜皂甙元。本文报道从中分离到的另一新三萜成分:3-hydroxy-12,14-diene-27-nordeane-28-nic acid。