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31.
Of 7 plasmids we tested, the plasmid pORF2 was eliminated in vitro with the most efficiency by treatment with subinhibitory concentrations of novobiocin, coumermycin and 10 quinolones. It showed a cure rate of 43% by enoxacin; 12% by novobiocin, pefloxacin, ciprofloxacin and CI-934; 7% by coumermycin and ofloxacin; 9% by amifloxacin; and 4% by AM-833. On the other hand, pSC194, pBR322 and pMH612 were poorly cured in vitro by quinolones, except pSC194 which was cured 33% by enoxacin. R1, pP1603, and pUB110 were unaffected by the treatment. Mice were challenged intraperitoneally with a 2XLD50 of Escherichia coli carrying the ORF2 plasmid and were treated per os with 1 X or 1/2 X ED50 of either enoxacin or CI-934. The frequency of loss of ampicillin resistance determined 3 h after treatment shows curing effects of 92% for CI-934, 89% for enoxacin and 20% for untreated control. 相似文献
32.
从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位 总被引:3,自引:1,他引:2
目的:建立一种有效从包涵体中纯化重组人幽门螺杆菌热休克蛋白A亚单位HspA的方法。方法:重组基因工程大肠杆菌发酵后,表达的Hp r HspA包涵体经洗涤,变性,复性,采用Q Sepharose High Performance阴离子交换层析和Superdex75凝胶过滤层析分离纯化,使用SDS-PAGE和HPLC检测纯度,选用ELISA和动物实验对纯化蛋白的免疫学活性和生物学活性进行鉴定。结果:Hp的HspA包涵经洗涤和溶解后,Hp的HspA的纯度>60%,包涵体溶解液经阴离子交换层析和凝胶过滤层析后纯度超过95%,Hp的HspA纯品经检测具有良好的免疫学活性和生物学活性。结论:本研究建立的分离纯化方法可从包涵体中获得高纯度的重组HspA蛋白,为进一步的动物实验研究奠定了基础。 相似文献
33.
Abra R. M. Hunt C. Anthony Fu K. K. Peters J. H. 《Cancer chemotherapy and pharmacology》1983,11(2):98-101
Cancer Chemotherapy and Pharmacology - Addition of solid doxorubicin or solutions to pre-formed liposomes proved to be the optimal method for incorporating the drug into liposomes whilst... 相似文献
34.
石景亮教授治疗多囊卵巢综合征的经验 总被引:3,自引:0,他引:3
石景亮(1939-),国家级第二批名老中医.业医三十余载,学验俱丰,对内科的脾胃病、肾病及妇科疑难杂症的治疗积累了丰富的经验.现就石老师治疗多囊卵巢综合征的经验介绍如下. 相似文献
35.
36.
c-myc is required for osteoclast differentiation. 总被引:1,自引:0,他引:1
R Battaglino D Kim J Fu B Vaage X Y Fu P Stashenko 《Journal of bone and mineral research》2002,17(5):763-773
The role of the receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL)-a tumor necrosis factor (TNF)-related cytokine-in osteoclast formation has been established clearly. However, the downstream signaling pathways activated by this cytokine remain largely unknown. To identify genes that play a role in osteoclastogenesis, we used RAW 264.7 mouse monocytes as a model system for the differentiation of multinucleated osteoclasts from mononucleated precursors. RAW 264.7 cells were induced with RANKL to form multinucleated giant osteoclast-like cells (OCLs) that expressed a number of osteoclast-specific markers and were able to form resorption pits on both calcium phosphate films and bone slices. This system was used to identify genes that are regulated by RANKL and may play a role in osteoclast differentiation. The proto-oncogene c-myc was strongly up-regulated in RANKL-induced OCLs but was absent in undifferentiated cells. Expression of Myc partners Max and Mad, on the other hand, was constant during OCL differentiation. We expressed a dominant negative Myc in RAW 264.7 cells and were able to block RANKL-induced OCL formation. Northern Blot analysis revealed a delay and a significant reduction in the level of messenger RNA (mRNA) for tartrate-resistant acid phosphatase (TRAP) and cathepsin K. We conclude that c-myc is a downstream target of RANKL and its expression is required for RANKL-induced osteoclastogenesis. 相似文献
37.
Wen-Ni Chang Jen-Ning Tsai Bing-Hung Chen Huei-Sheng Huang Tzu-Fun Fu 《Drug metabolism and disposition》2007,35(11):2127-2137
Serine hydroxymethyltransferase (SHMT) provides activated one-carbon units required for the biosynthesis of nucleotides, protein, and methyl group by converting serine and tetrahydrofolate to glycine and N(5),N(10)-methylenetetrahydrofolate. It is postulated that SHMT activity is associated with the development of methotrexate resistance and the in vivo activity of SHMT is regulated by the binding of N(5)-CHO-THF, the rescue agent in high-dose methotrexate chemotherapy. The aim of this study is to advance our understanding of the folate-mediated one-carbon metabolism in zebrafish by characterizing zebrafish mitochondrial SHMT. The cDNA encoding zebrafish mitochondrial SHMT was cloned, overexpressed in Escherichia coli, and purified with a three-step purification protocol. Similarities in structural, physical, and kinetic properties were revealed between the recombinant zebrafish mitochondrial SHMT and its mammalian orthologs. Surprisingly, leucovorin significantly inhibits the aldol cleavage of serine catalyzed by zebrafish cytosolic SHMT but inhibits to a lesser extent the reaction catalyzed by the mitochondrial isozyme. This is, to our knowledge, the first report on zebrafish mitochondrial folate enzyme as well as the differential inhibition of leucovorin on these two SHMT isoforms. Western blot analysis revealed tissue-specific distribution with the highest enrichment present in liver for both cytosolic and mitochondrial SHMTs. Intracellular localization was confirmed by confocal microscopy for both mitochondrial and cytosolic SHMTs. Unexpectedly, the cytosolic isoform was observed in both nucleus and cytosol. Together with the previous report on zebrafish cytosolic SHMT, we suggest that zSHMTs can be used in in vitro assays for folate-related investigation and antifolate drug discovery. 相似文献
38.
Yulu Miao Mingxia Zhang Yulin Nie Wan Zhao Bin Huang Zhengming Jiang Shaoxiong Yu Zhibin Huang Hongjin Fu 《中国神经再生研究》2007,2(2):126-128
BACKGROUND: Besides local changes of cranial parenchymal cells, hemorrhage, etc., severe traumatic brain injuries also cause the changes of total body fluid and various functions, and the changes of lymphocytes and T lymphocyte subsets should be paid more attention to. OBJECTIVE: To reveal the changing laws of T lymphocyte subsets after severe traumatic brain injury, and compare with mild to moderate brain injury. DESIGN: A comparative observation. SETTINGS: Department of Neurosurgery, Longgang District Buji People's Hospital of Shenzhen City; Central Laboratory of Shenzhen Hospital of Prevention and Cure for Chronic Disease. PARTICIPANTS: All the subjects were selected from the Department of Neurosurgery, Longgang District Buji People's Hospital of Shenzhen City from August 2002 to August 2005. Thirty patients with severe brain injury, whose Glasgow coma score (GCS) was ≤ 8 points, were taken as the experimental group, including 21 males and 9 females, aging 16 - 62 years. Meanwhile, 30 patients with mild traumatic brain injury were taken as the control group (GCS ranged 14- 15 points), including 18 males and 12 females, aging 15 -58 years. All the subjects were in admission at 6 hours after injury, without disease of major organs before injury Informed consents were obtained from all the patients or their relatives. METHODS: (1) The T lymphocytes and the subsets in peripheral blood were detected with immunofluorescent tricolor flow cytometry at l, 3, 7 and 14 days after injury in both groups. (2) The conditions of pulmonary infections were observed at 4 days after injury. The differences of measurement data were compared with the t test. MAIN OUTCOME MEASURES: Changes of T lymphocytes subsets at 1 - 14 days after severe and mild or moderate traumatic injury. RESULTS: Finally, 28 and 25 patients with mild to moderate traumatic brain injury, whereas 25 and 21 patients with severe traumatic brain injury were analyzed at 7 and 14 days respectively, and the missed ones died due to the development of disease. (1) Changes of T lymphocyte subsets: At 1 and 3 days after injury, CD3, CD4, CD8, CD4/CD8 began to decrease, whereas CD8 increased in the experimental group, which were very significantly different from those in the control group (t =2.77 - 3.26, P 〈 0.01), and began to recover at 7 days, which were significantly different from those in the control group (t = 2.06 - 2.24, P 〈 0.05), and generally recovered to the normal levels at 14 days (P 〉 0.05). (2) Conditions of pulmonary infections: At 4 days after injury, the rate of pulmonary infection was significantly different between the experimental group and control group [73% (22/30), 0, x2=37.29, P 〈 0.01]. CONCLUSION: Patients with severe traumatic brain injury suffer from damages of cellular immune function at early period (within 7 days), and they are easily to be accompanied by pulmonary infections. 相似文献
39.
以钒基核苷酸复合物为核酸酶抑制剂,从人胚组织中制备总RNA,所得产品几乎不含DNA和蛋白质,经过分离Poly(A)~ mRNA,证明制品有合成cDNA的完整功能,方法比较简捷,适于大量制备以满足分离mRNA 之需,并讨论了此法的优缺点。 相似文献
40.
H N Qian J Feng H Cui B S Gao S L Qin T Y Fu P Wei Z Y Fu S M Gao Z Y Yang 《中华医学杂志(英文版)》1991,104(2):109-113
Monoclonal antibody 131I-COC183B2, developed in our laboratory and proved to fit for human treatment was injected intraperitoneally or subcutaneously in 13 patients. In 8 cases with i.p. injection the disease corresponded with the image, i.e. 3 primary ovarian epithelial cancers showed positive images, 1 ovarian Krukenberg tumor was negative and the other 4 negative images included 1 uterine myoma and 3 ovarian teratomas. In the subcutaneous injection group, 4 cases had ovarian carcinoma, surgery and chemotherapy. Two negative images corresponded with the clinical status-in good health, another negative case had metastatic left supraclavicular lymph node due to ovarian mucinous adenocarcinoma. The last negative image in this group was a case of benign ovarian teratoma which was proved after surgery. The 1 positive case was waiting to be proved by a scheduled third operation. The computer scintigram calculation of T/NT was 5.35 to 13.7. The results suggest that this monoclonal antibody can be used for radioimmunoimaging for the localization of ovarian carcinoma, which is not only helpful for clinical staging and differential diagnosis but is also a good follow-up method.
相似文献