1.Study on correlation of staining method and contamination.
Xinlan LUO ; Jun YAO ; Yongjun HUANG ; Kunming LIANG ; Xiaolan ZHU ; Wenxian DOU ; Xuefeng LIU ; Lijuan KUANG ; Lingyin LIANG ; Yanhui LIU
Chinese Journal of Pathology 2014;43(12):834-836
4.Screening for hyper-accumulating lipid mutants in Aurantiochytrium limacinum using high-throughput fluorescence-based method.
Guangqian DUAN ; Shuoshuo LI ; Xin LI ; Kaiyao HUANG
Chinese Journal of Biotechnology 2019;35(7):1335-1347
Docosahexaenoic acid (DHA) has many unique physiological functions such as promoting the development of brain and retina in infants. Therefore, it is widely applied to food, pharmacy, breeding and other industries. To obtain engineered strains of Aurantiochytrium limacinum SR21 suitable for industrial application with increased lipid and DHA production, we designed a simple, fast, accurate and high-throughput screening method based on Nile red staining of oil droplets. First, ultraviolet C (UVC) mutagenesis was used to generate a random mutant library of A. limacinum. Second, screening conditions were optimized including staining conditions of Nile red and the sorting criterion. Thereby, three putative high-lipid mutants (D03432, D05106 and D01521) were selected from the mutant library containing 3 648 mutated clones. The three mutants grew faster and produced higher amounts of lipids and DHA compared to wild type (WT). In 100 mL cultures, the lipid content of D03432 and D05106 mutants reached 64.74% and 63.13% of dry cell weight respectively, whereas the wild strain exhibited only 43.19%. DHA yield in these two mutants were even 2.26-fold and 2.37-fold higher than that of the wild strain. Experiment with 5 L fermentor further confirmed that D03432 and D05106 mutants had better performance than the wild strain on DHA yield (45.51% and 66.46% more than that of the wild strain, respectively), and demonstrated their high potential for industrial application. This work not only generated several high-DHA content mutants with high potential for industrial use, but also provided vital guidance for high-throughput screening of lipid hyper-accumulating mutants in other oil-producing microorganisms.
Bioreactors
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Docosahexaenoic Acids
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Mutagenesis
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Staining and Labeling
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Stramenopiles
5.Improvement program on pretreatment of acid decalcified tissue in hematoxylin-eosin staining.
Li-Hong YAO ; Mei ZHANG ; Mei-Chang HUANG ; Zi-Xin WAN ; Wei-Long ZHANG ; Xiao YANG ; Ming-Zhong YANG ; Yu CHEN ; Ya-Ling TANG
West China Journal of Stomatology 2020;38(3):297-300
OBJECTIVE:
To explore the treatment conditions of acid decalcified specimens and improve the poor quality of sections and unclear structure of hematoxylin-eosin (HE) staining caused by the change in pH in tooth and hard tissue after acid decalcification.
METHODS:
A total of 20 cases of oral pathological specimens that contain hard tissues were decalcified and treated with routine treatment, concentrated ammonia water immersion treatment, and saturated lithium carbonate solution immersion treatment. The quality and HE staining effects of hard tissue sections treated with different methods were compared.
RESULTS:
Compared with routine treatment, lithium carbonate saturated solution treatment showed complete sections. Hematoxylin is strongly stained, the nucleus is clear, and the cytoplasm is bright.
CONCLUSIONS
Soaking acid decalcified specimens in lithium carbonate saturated solution before embedding in dehydration can neutralize the acidic environment of the tissue. The quality of sections and HE staining effect are improved and are suitable for the pretreatment of acid decalcified tissue samples of oral pathology.
Eosine Yellowish-(YS)
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Hematoxylin
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Staining and Labeling
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Tooth
7.Hepatocellular Carcinoma of the Liver with Sarcomatous Feature.
Dong Hoon SHIN ; Won Geun KANG ; Mung Hi YOON ; Bong Kwon CHUN ; Keyng Seung OH ; Choong Han LEE
Korean Journal of Hepato-Biliary-Pancreatic Surgery 2001;5(2):113-117
One case of hepatocellular carcinoma with sarcomatous changes is presented. Histologically, the tumor was composed of hepatocellular-carcinomatous and sarcomatous components, including trabecular, pseudoglandular, and spindle-shaped varieties. There was a transitional cell form between the carcinoma and sarcomatous cells. Immunohistochemical examination for alpha-feto protein reveals positive staining in hepatocellular carcinoma component, and vimentin had positive result in most spindle-shaped sarcomatous cells and some epithelial tumor cells, whereas cytokeratin and EMA(epithelial membrane antigen) revealed negative staining in tumor cells, On the basis of this findings, the possibility of sarcomatous transformation of hepatocellular carcinoma was discussed.
Carcinoma, Hepatocellular*
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Immunohistochemistry
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Keratins
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Liver*
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Membranes
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Negative Staining
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Vimentin
10.Application of experion(TM) assay system for discovery of ovarian cancer serum biomarkers.
Hyun Jin MIN ; Su Mi BAE ; Sun Yong KWAK ; Min Ah YANG ; Hae Nam LEE ; Yong Wook KIM ; Jong Chul SHIN ; Jung Sick KIM ; Kye Hyun NAM ; Yong Wan KIM ; Woong Shick AHN
Korean Journal of Obstetrics and Gynecology 2007;50(5):751-759
OBJECTIVE: The discovery of new biomarkers for ovarian cancer is clearly necessary for the detection and monitoring of the disease. Experion(TM) automated electrophoresis system can be employed in the identification of differentially expressed proteins in cancer cells. The objective of this study was to discover potential diagnostic serological biomarkers for ovarian cancer. METHODS: We performed protein expression difference analyses for 14 healthy women and 28 ovarian cancer patients with stage I, III and IV using Experion(TM) system. And then we checked the protein expression as silver staining after loading at 8~16% gradient gel for comparison with Experion(TM) gel image. The candidate biomarkers were purified and determined using MALDI-TOF mass spectrometer. RESULTS: The distinctive polypeptide peaks were detected at 115.40, 15.96, 14.8, 11.66, and 10.69 kDa and these five peaks were identified as ceruloplasmin, hemoglobin beta chain, hemoglobin sigma chain, serum amyloid A4, and amyloid related serum protein SAA, respectively. These proteins were significantly different between the sera of normal healthy women and ovarian cancer patients. CONCLUSIONS: Five proteins were found to be significantly different between the sera of normal healthy women and ovarian cancer patients. In addition, Experion(TM) assay system can provide high performance for analysis of ovarian cancer-related proteins by increasing the throughput while maintaining a high level of accuracy.
Amyloid
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Biomarkers*
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Ceruloplasmin
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Electrophoresis
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Female
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Humans
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Ovarian Neoplasms*
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Silver Staining