3.Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites
Hong-Bing MA ; Yu-Hua QIU ; Ran TAO ; Wen-Xiang LI ; Ying XU ; Xue-Guang ZHANG ;
China Biotechnology 2006;0(08):-
A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.
5.Clinicopathological analysis of myelolipoma within adrenocortical adenoma
Wei QIU ; Ye HONG ; Rusong ZHANG ; Xue WEI ; Zhenfeng LU ; Qunli SHI ; Jie MA
Journal of Medical Postgraduates 2017;30(6):637-640
Objective At present, there are few studies about myelolipoma within adrenal cortical adenoma.Our aim was to provide more basis for correct diagnosis and treatment by investigation into its clinical and pathological features.Methods The clinical and pathological data were retrospectively reviewed in 11 patients of myelolipoma within adrenal cortical adenoma, along with relative literature reviews.Results The median age of 11 patients (7 females, 4 males) was 49±9.5 years, among whom 3 patients presented Cushing's syndrome, 1 patient with more than 10 years' recurrent dizzy with hypertension, other 7 patients were found coincidently by routine examination.Adrenal mass were found by imaging examination.Pathologically, myelolipomas were in solitary nodule distribution and/or admixed with adrenal cortical adenomas.Myelolipomas were composed of variable admixture of mature adipose tissue and hematopoietic elements.Surgical treatment was performed for all 11 patients, and no relapse was found in 2 months' to 11 years' follow-up.Conclusion Myelolipoma within adrenal cortical adenoma is extremely rare, which is common in females.The patients may present with Cushing's syndrome, hypertension or without obvious clinical syndrome.All the patients are in favorable prognosis after surgical resection.
6.Relationship between efficacy exertion of diuretic traditional Chinese medicines and aquaporin.
Peng-cheng WANG ; Shan ZHAO ; Qiu-hong WANG ; Hai-xue KUANG
China Journal of Chinese Materia Medica 2015;40(12):2272-2277
In recent years, the discovery and studies on aquaporin have made us have a more in-depth understanding about the physiological and pathological processes of water metabolism. Over years, however, there has been no quantitative study on the target sites of diuretic traditional Chinese medicines at the molecular level. In that case, aquaporin was found to been a new target molecule to explain the efficacy exertion of diuretic traditional Chinese medicines. By studying aquaporin, researchers can understand the implicit meaning of the diuretic effect of traditional Chinese medicines and conduct quantitative studies on the diuretic effect. So far, many scholars have conducted a series of studies in the traditional Chinese medicine field by using the findings on aquaporin and made certain advances. This article provides a summary about the efficacy exertion of diuretic traditional Chinese medicines through target molecule aquaporin.
Animals
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Aquaporins
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genetics
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metabolism
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Diuretics
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pharmacology
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Medicine, Chinese Traditional
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Water
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metabolism
7.Strong Expression of Recombinant Human Morphogenetic Protein-4 in Escherichia coli and its Bioassay in vivo
Si-Hong GAO ; Ju WANG ; Que-Wei DONG ; Kan LIU ; Xue-Ting LIU ; An HONG ; Qiu-Ling XIE ; Fen-Yong SUN ;
China Biotechnology 2006;0(03):-
Objective:To produce rhBMP-4 with bioactivity in E.coli. Methods: The full-length human BMP-4 gene was mutated by PCR without changes in amino acid sequence, then the synthesized gene was cloned into plasmid pET-3c, transducted into BL21(DE)plysS, and induced by adding IPTG to a final concentration of 1.0 mmol/L. The protein product was purified using ion-exchange chromatography method and then renaturated, bioactivity was checked by C2C12 differentiation in vitro and mouse ectopic bone formation in vivo. Results: A 438 bp gene fragment encoding mature peptide of hBMP-4 was cloned , the protein product was mostly in the form of inclusion body, after renaturation, the engineering protein shows better bioactivity. Conclusion:The mutant strategy can enhance the expression of bioactive rhBMP-4 in E.coli expression system.
8.Expression of human epididymis protein 2beta1 in the testis and epididymis of adolescent male rats.
Hong TIAN ; Shu-dong QIU ; Qiu-yang ZHANG ; Xia XUE ; Ling GE ; Li-rong WANG
National Journal of Andrology 2006;12(9):780-783
OBJECTIVETo investigate the expression of human epididymal secretary protein 2 isoform human epididymal protein 2beta1(HE2beta1) in the testis and epididymis of adolescent male rats along with its significance.
METHODSImmunohistochemical staining was used to detect the expression and localization of HE2beta1 in the testis and epididymis of 15 adolescent SD rats.
RESULTSHE2beta1 immunoreactive staining was detected in the testis and epididymis. In the epithelia of the epididymal duct, HE2beta1 expressed mainly in the supranuclear region of the principle cells and the basement membrane of some epithelial cells; there were no immunostaining in the n clear cells, halo cells and basal cells. The immunopositive reaction was detected, weak in the distal caput, strong in the proximal, middle corpus and the cauda, but negative in the initial segment. Immunopositive results of HE2beta1 were also observed in some of the nuclei of spermatogonia and Sertoli cells with negatively-stained cytoplasm.
CONCLUSIONImmunohistochemical staining is a fairly sensitive method for detecting HE2beta1 expression. The localization and expression level of HE2beta1 in the genital duct of adolescent male rats exhibited a region- and cell-specific expression pattern, which suggests that HE2beta1 may play an important role in spermatogenesis, maturation and epididymal epithelial innate defense mechanisms.
Animals ; Antigens, Surface ; biosynthesis ; Epididymis ; metabolism ; Glycopeptides ; biosynthesis ; Humans ; Immunohistochemistry ; Leydig Cells ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Testis ; metabolism
9.Time-series Analysis in Imatinib-resistant Chronic Myeloid Leukemia K562-cells under Different Drug Treatments
ZHAO YAN-HONG ; ZHANG XUE-FANG ; ZHAO YAN-QIU ; BAI FAN ; QIN FAN ; SUN JING ; DONG YING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):621-627
Chronic myeloid leukemia (CML) is characterized by the accumulation of active BCR-ABL protein.Imatinib is the first-line treatment of CML;however,many patients are resistant to this drug.In this study,we aimed to compare the differences in expression patterns and functions of time-series genes in imatinib-resistant CML cells under different drug treatments.GSE24946 was downloaded from the GEO database,which included 17 samples of K562-r cells with (n=12) or without drug administration (n=5).Three drug treatment groups were considered for this study:arsenic trioxide (ATO),AMN107,and ATO+AMN107.Each group had one sample at each time point (3,12,24,and 48 h).Time-series genes with a ratio of standard deviation/average (coefficient of variation) >0.15 were screened,and their expression patterns were revealed based on Short Time-series Expression Miner (STEM).Then,the functional enrichment analysis of time-series genes in each group was performed using DAVID,and the genes enriched in the top ten functional categories were extracted to detect their expression patterns.Different time-series genes were identified in the three groups,and most of them were enriched in the ribosome and oxidative phosphorylation pathways.Time-series genes in the three treatment groups had different expression patterns and functions.Time-series genes in the ATO group (e.g.CCNA2 and DAB2)were significantly associated with cell adhesion,those in the AMN107 group were related to cellular carbohydrate metabolic process,while those in the ATO+AMN107 group (e.g.AP2M1) were significantly related to cell proliferation and antigen processing.In imatinib-resistant CML cells,ATO could influence genes related to cell adhesion,AMN107 might affect genes involved in cellular carbohydrate metabolism,and the combination therapy might regulate genes involved in cell proliferation.
10.Inhibitory effect of melatonin on morphine withdrawal syndromes and the content of NO in plasma and brain tissue in morphine dependent mice.
Ying-hong ZHOU ; Zhan-yang HUO ; Xue-cai QIU
Acta Pharmaceutica Sinica 2002;37(3):175-177
AIMTo observe the effect of melatonin (MT) on morphine withdrawal syndromes and determine the content of NO in plasma and brain tissue in morphine dependent mice.
METHODSA physical dependent model in mice was established by subcutaneous injection of morphine. MT (15 mg.kg-1, qd x 3) was given by intragastric infusion (ig) for three days. Withdrawal syndromes were induced by intraperitoneal injection of naloxon (5 mg.kg-1). The intensity of withdrawal syndromes was evaluated according to the jumping latency, the jumping times and the body weight loss. The content of NO was detected with Griess method.
RESULTSThe jumping latency of morphine withdrawal reaction was prolonged and the jumping times were reduced obviously by ig MT. The increased NO content in plasma and brain tissue in morphine dependent mice was reduced by ig MT.
CONCLUSIONThe physical withdrawal syndromes and the content of NO in plasma and brain tissue in morphine dependent mice are inhibited by MT.
Animals ; Brain ; drug effects ; metabolism ; Disease Models, Animal ; Male ; Melatonin ; therapeutic use ; Mice ; Morphine Dependence ; blood ; Nitric Oxide ; blood ; Substance Withdrawal Syndrome ; blood ; prevention & control