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排序方式: 共有195条查询结果,搜索用时 31 毫秒
151.
Wang L Ma W Zhu L Ye D Li Y Liu S Li H Zuo W Li B Ye W Chen L 《American journal of physiology. Cell physiology》2012,303(1):C14-C23
Acid-activated chloride currents have been reported in several cell types and may play important roles in regulation of cell function. However, the molecular identities of the channels that mediate the currents are not defined. In this study, activation of the acid-induced chloride current and the possible candidates of the acid-activated chloride channel were investigated in human nasopharyngeal carcinoma cells (CNE-2Z). A chloride current was activated when extracellular pH was reduced to 6.6 from 7.4. However, a further decrease of extracellular pH to 5.8 inhibited the current. The current was weakly outward-rectified and was suppressed by hypertonicity-induced cell shrinkage and by the chloride channel blockers 5-nitro-2-3-phenylpropylamino benzoic acid (NPPB), tamoxifen, and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid disodium salt hydrate (DIDS). The permeability sequence of the channel to anions was I(-) > Br(-) > Cl(-) > gluconate(-). Among the ClC chloride channels, ClC-3 and ClC-7 were strongly expressed in CNE-2Z cells. Knockdown of ClC-3 expression with ClC-3 small interfering (si)RNA prevented the activation of the acid-induced current, but silence of ClC-7 expression with ClC-7 siRNA did not significantly affect the current. The results suggest that the chloride channel mediating the acid-induced chloride current was volume sensitive. ClC-3 is a candidate of the channel proteins that mediate or regulate the acid-activated chloride current in nasopharyngeal carcinoma cells. 相似文献
152.
153.
A novel evodiamine (EVO)-phospholipid complex (EPLC) was designed to improve the bioavailability of EVO. A central composite design approach was employed for process optimization. EPLC were characterized by differential scanning calorimetry, ultraviolet spectroscopy, Fourier transformed infrared spectroscopy, 1H-NMR spectroscopy, matrix-assisted laser desorption/ionization time-of-flight spectroscopy, apparent solubility, and dissolution rate. After oral administration of EPLC, the concentrations of EVO at different time points were determined by high-performance liquid chromatography. The optimal formulation for EPLC was obtained where the values of X1, X2, and X3 were 2, 0.5, and 2.5 mg/mL, respectively. The average particle size and zeta potential of EPLC with the optimized formulation were 246.1 nm and −26.94 mV, respectively. The EVO and phospholipids in the EPLC were associated with non-covalent interactions. The solubility of EPLC in water and the dissolution rate of EPLC in phosphate-buffered solution (pH 6.8) were substantially enhanced. The plasma EVO concentration-time curves of EPLC and free EVO were both in accordance with the two-compartment model. The peak concentration and AUC0−∞ of EPLC were increased, and the relative bioavailability was significantly increased to 218.82 % compared with that of EVO.KEY WORDS: bioavailability, evodiamine, phospholipid complex, process optimization 相似文献
154.
盐霉素(salinomycin)是由白色链霉菌(Streptomyces albus)产生的一元羧酸聚醚类抗生素,具有较强的抗革兰氏阳性菌和杀灭球虫的作用,而且对环境污染也较低;此外,还能特异性地抑制多种肿瘤细胞及肿瘤干细胞的生长,具有多重作用靶点,有望成为抗肿瘤的特效药。为提高盐霉素的产量,人们采用传统诱变技术和现代分子遗传学手段,对盐霉素产生菌进行了改造,获得了高产菌株;同时,通过对盐霉素化学结构进行修饰或者通过药物载体和联合用药等,增强了其活性和靶向性、减少了毒副作用。本文对盐霉素产生菌的改造策略、药物靶向性提高和活性优化等研究进展进行综述,并对今后的研究热点进行展望。 相似文献
155.
【目的】由于谷氏菌素产生菌禾粟链霉菌的遗传操作效率较低,本研究在谷氏菌素生物合成基因簇成功异源表达的基础上,通过在异源宿主天蓝色链霉菌M1146中阻断谷氏菌素生物合成基因gouC和gouD,研究其在谷氏菌素生物合成中的作用,为谷氏菌素生物合成途径的阐明奠定基础。【方法】以含有谷氏菌素生物合成基因簇的黏粒D6-4H为基础,通过PCR-targeting的方法分别将gouC和gouD敲除得到重组质粒p GOUe-ΔC和p GOUe-ΔD。通过接合转移将这两个重组质粒分别导入天蓝色链霉菌M1146中,获得gouC和gouD的缺失突变株M1146-GOUe-ΔC和M1146-GOUe-ΔD,通过HPLC分析突变株的谷氏菌素中间产物积累情况,分离纯化后对其进行结构鉴定和生物活性检测。【结果】gouC和gouD的缺失均导致谷氏菌素不能合成,突变株发酵液中积累了不同的中间产物,生物活性分析发现这些中间产物均失去了对肿瘤细胞的抑制活性。【结论】gouC和gouD是谷氏菌素生物合成的重要基因,与谷氏菌素肽基部分的肌氨酸残基合成相关。本研究为阐明谷氏菌素的生物合成机制提供了更多的依据。 相似文献
156.
157.
Characterization of the Polyoxin Biosynthetic Gene Cluster from
Streptomyces cacaoi and Engineered Production of Polyoxin
H 总被引:1,自引:0,他引:1
158.
Shuli Zhang Bo Liu Haihua Yang Yuqing Tian Gang Liu Lei Li Huarong Tan 《Nucleic acids research》2010,38(11):3709-3720
EndoTT encoded by tte0829 of Thermoanaerobacter tengcongensis binds and cleaves single-stranded (ss) and damaged double-stranded (ds) DNA in vitro as well as binding dsDNA. In the presence of a low concentration of NaCl, EndoTT cleaved ss regions of damaged dsDNA efficiently but did not cleave DNA that was entirely ss or ds. At high concentrations of NaCl or MgCl2 or ATP, there was also specific cleavage of ssDNA. This suggested a preference for ss/ds junctions to stimulate cleavage of the DNA substrates. EndoTT has six specific sites (a–f) in the oriC region (1–70 nt) of T. tengcongensis. Substitutions of nucleotides around site c prevented cleavage by EndoTT of both sites c and d, implying that the cleavage specificity may depend on both the nucleotide sequence and the secondary structure of the ssDNA. A C-terminal sub-fragment of EndoTT (residues 107–216) had both endonucleolytic and DNA-binding activity, whereas an N-terminal sub-fragment (residues 1–110) displayed only ssDNA-binding activity. Site-directed mutations showed that G170, R172 and G177 are required for the endonuclease activity of EndoTT, but not for DNA-binding, whereas D171, R178 and G189 are partially required for the DNA-binding activity. 相似文献
159.
Li H Oppert B Higgins RA Huang F Buschman LL Gao JR Zhu KY 《Insect biochemistry and molecular biology》2005,35(8):847-860
Our previous studies suggested that Bacillus thuringiensis (Bt) resistance in a Dipel-resistant strain of Ostrinia nubilalis was primarily due to reduced trypsin-like proteinase activity. In this study, we demonstrated a 254-fold resistance to Cry1Ab protoxin but only 12-fold to trypsin-activated Cry1Ab toxin in the Dipel-resistant strain. Significantly higher resistance to Cry1Ab protoxin than to trypsin-activated Cry1Ab toxin further supports the hypothesis that reduced trypsin-like proteinase activity leading to reduced activation of the Bt protoxin is a major resistance mechanism in the Dipel-resistant strain. To understand the molecular basis of reduced proteinase activity, three cDNAs, OnT2, OnT23, and OnT25, encoding full-length trypsin-like proteinases, were sequenced in Bt-resistant and -susceptible O. nubilalis larvae. Although a number of nucleotide differences were found in sequences from the Bt-resistant and -susceptible strains, the differences were not consistent with reduced trypsin-like activity in the Bt-resistant strain. However, the mRNA levels of OnT23 in the resistant strain were 2.7- and 3.8-fold lower than those of the susceptible strain as determined by northern blotting and real-time quantitative PCR, respectively. Thus, reduced trypsin-like activity may be attributed to reduced expression of OnT23 in Bt-resistant O. nubilalis. Our study provides new insights into Bt resistance management strategies, as resistance mediated by reduced Bt protoxin activation would be ineffective if resistant insects ingest a fully activated form of Cry1Ab toxin, either in spray formulations or transgenic Bt crops. 相似文献
160.
A food-grade gene expression system in Lactococcus lactis was established by the combination of a vector containing the lacF gene as the selection marker and a strain WZ103 carrying an in-frame deletion of this gene in the chromosome as the host. The human glutathione S-transferase A1-1 (hGSTA1) and Cu/Zn superoxide dismutase (hSOD) genes were respectively cloned into a food-grade vector under the control of the lactococcal inducible promoter P(lacA). The resulting expression plasmids were separately introduced into the lactose-deficient (Lac(-)) host, and the lactose-utilizing (Lac(+)) transformants were directly selected on a chemically defined medium, using lactose as the sole carbon source. The successful food-grade expression of hGSTA1 and hSOD in the L. lactis WZ103 transformed with these plasmids were analyzed by Western blotting and enzymatic activity assay, respectively. 相似文献