1.Cloning expression purification and cytotoxic assay of sTRAIL in A549 cell line
Chinese Pharmacological Bulletin 1987;0(02):-
Aim To clone,express sTRAIL gene,then purify sTRAIL(114-281 amino acid) and assay its cytotoxic activity in human A549 cell line.Methods The intact full human TRAIL gene was amplified using PCR method from the human placenta and lung cDNA library.The full human TRAIL cDNA gene was inserted into pUC19 vector and sequenced.The extracellular DNA fragment was amplified using PCR,which was cloned to pET-11a vector and transformed into E.coli BL21.The denatured and refolded sTRAIL was purified and cytotoxic activity of sTRAIL was assayed using crystal violet staining and fluorescence-activated cell sort(FACS) in A549 cell line.Results The full length TRAIL cDNA gene was amplified from the human placenta and lung cDNA library,which was identical to the published TRAIL sequence.The extracellular DNA fragment was cloned to pET-11a.The expression level reached 50% of the total protein of BL21.The purity of sTRAIL was about 98%,while IC_(50) was about(24?5.2) ?g?L~(1) in TRAIL-treated A549 cells with crystal violet staining method.The time-dependent relationship of sTRAIL-induced apoptotic death in A549 cells was significant with FASC analysis.Conclusion sTRAIL gene has been cloned and successfully expressed.The process of refolding and purification of sTRAIL has been established.sTRAIL demonstrated cytotoxicity in A549 cell line.
2.Research Progress in Genotoxic Effects of Degradation Products, Cobalt, Chromium Ions and Nanoparticles from Metal-on-metal Prostheses on Cells.
Hao ZHOU ; Qinglin HAN ; Fan LIU
Journal of Biomedical Engineering 2015;32(2):489-492
Cobalt or chromium alloys are the most common clinical materials of prosthesis and there have been some investigators at home and abroad have done related researches about the genotoxic effects of cobalt and chromium ions and nanoparticles. People have certain understanding about the mechanism of production of ions as well as their influence on cells. However, chromium or cobalt nanoparticles genotoxicity related research is still in its preliminary stage. In each stage, the mechanisms, from creating of the particles, through entering cells, until finally causing genotoxic, are still contained many problems to be solved. This article reviews the research progress in mechanisms of production and genotoxic effects of cobalt, chromium ions and nanoparticles.
Chromium
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toxicity
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Cobalt
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toxicity
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DNA Damage
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Humans
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Ions
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Nanoparticles
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toxicity
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Prostheses and Implants
3.Progress and clinical application of TRAIL-induced apoptosis in cancer
Qinglin FAN ; Lihua SONG ; Wei WEI ;
Chinese Pharmacological Bulletin 2003;0(09):-
TNF related apoptosis inducing ligand (TRAIL) is a new identified member of TNF family, which selectively kills a broad spectrum of tumor cells, but nontoxic to most normal cells. TRAIL triggers tumor cell apoptosis via death receptor DR4 and DR5 anchored in the cell surface, which is mediated through intracellular induced apoptosis proteins. The major pathway of its action proceeds through the formation of DSIC and activation of caspase8. The apoptotic processes follow two signal pathways, namely the mitochondrial independent activation of caspase3, and mitochondrial dependent apoptosis through the cleavage of BID by caspase8, the release of cytochrome C, the formation of apoptosome, and activation of caspase9 and the downstream apoptosis. In previous researches, it is shown that nontagged TRAIL proved to be noncytotoxic to hepatocytes in monkeys and chimpanzees and to human normal hepatocytes. Thus, the nontagged TRAIL has attracted great attention in recent years as a promising anti cancer reagent.
4.Reconstruction of bone and skin defect at medial malleolns
Qingluan HAN ; Wei LI ; Hongjin FAN ; Qinglin ZHANG ; Zhi ZHANG
Chinese Journal of Microsurgery 2015;38(4):354-358
Objective To investigate the operative method and clinical rusults of repairing and reconstruction for bone and skin defect at medial malleolus.Methods Form January,2013 to January,2015,11 cases of patients with malleolus complex tissue defect were treated.According to the degree of damage to the cases were divided into four types,according to the type,selected flap,blood vessels,nerves,tendon graft,flap series connection iliac bone flap graft to repair,or direct ankle fusion,amputation to therapy.The Iliac flap donor site remained part of the anterior superior iliac spine and rebuild muscle starting and ending point,the donor sites wound was closed by skin graft.Results Followed-up of 1-24 months,in addition to amputation and ankle fusion each one exception,the remaining 9 patients underwent ankle reconstruction to reserve ankle.One case acquired infection,other 8 patients healed well,had different degree of recovery of the ankle function,the ankle function had recovery in different degree.AOFAS ankle-hindfoot score system:excellent in 4 cases,good in 4 cases,and poor in 1 case.Conclusion Ankle complex tissue defect classification method can be used to guide us to select the treatment options.Iliac bone flap series consisting mainly of composite tissue transplantation is a viable method of the medial malleolus defect reconstruction.
5.Reteplase fusion protein:expression, purification and the effect of chaperones on its renaturation
Wei WANG ; Wenyi ZOU ; Qinglin FAN ; Lihua SONG
Acta Universitatis Medicinalis Anhui 2013;(11):1287-1291
Objective To construct the prokaryotic expression system of reteplase fusion protein and research the effect of chaperones on its renaturation. Methods Inserted the reteplase gene into the prokaryotie expression vector PET32a and then expressed it by the induction of IPTG in E. coli BL21. Researched the effect of chaperones on the renaturation of fusion protein by adding different chaperones. Results The analysis of SDS-PAGE and Western blot indicated that reteplase fusion protein was expressed correctly. Chaperones DsbA,pKJE7,pTf16 had the conspicu-ous effect on the renaturation of fusion protein. The result of activity assay indicated that the refolded reteplase fu-sion protein had fibrinolytic activity. Conclusion Chaperones can promote renaturation of reteplase fusion protein.
6.Research progress of the anti-hepatitis virus mechanism of interferon-alpha and long-acting form of interferons
Bingfa XU ; Qinglin FAN ; Wei WEI ; Lihua SONG
Chinese Pharmacological Bulletin 2003;0(10):-
Interferon-alpha is one of the most important drugs against hepatitis virus,and it can inhibit the replication of virus by inducing the expression of 2'-5'-OAS,PKR,MxA through Jak-STAT pathway in hepatocytes.The effects are also concerned with other signal pathways,such as MAPK,IRS-1/PI3K-p70S6 kinase pathways and so on.In order to prolong the half life of interferon-alpha and enhance its effectiveness,many other long-lasting interferons are developed,such as pegylated interferon,recombinant human serum albumin-interferon,and liposome interferon,which maybe take the place of interferon-alpha.The research and development of more long-acting and efficient interferons are becoming a more and more important way to the anti-virus treatment of hepatitis.
7.Expression and relationship of Beclin1 and Bci2 in invasive pituitary adenomas
Zhuguo RAN ; Qinglin FENG ; Yi SONG ; Jiangfeng DU ; Mingdong LIU ; Shibing FAN ; Ji LI ; Gang HUO ; Liuyang WU ; Gang YANG ; Rui ZHAO ; Mei FENG ; Kun TIAN ; Xiuhua HAN
Journal of Endocrine Surgery 2012;06(4):253-256
Objective To detect the expression of Beclin1 and Bcl2 in invasive pituitary adenomas and to explore the relationship of Beclin1 and Bci2 in invasive pituitary adenomas and the relativity between the 2 genes.Methods 61 specimens were classified into invasive group (32 cases) and non-invasive group (29 cases) according to the comprehensive evaluation of invasive pituitary adenomas.lmmunofluorescence analysis and RT-PCR were adopted respectively to detect the protein and mRNA expressions of Beclinl and Bcl2.The difference and relativity of Beclin1 and Bcl2 expression in invasive group and non-invasive group were analyzed.Results 32 specimens of pituitary adenoma were invasive and 29 were non-invasive.Beclin1 protein and mRNA expressions were lower in the invasive group than in the non-invasive group (P <0.01 ).Bcl2 protein and mRNA expressions were higher in the invasive group than in the non-invasive group (P <0.01 ).Pearson related analysis showed that Beclin1 mRNA expression was negtively correlated with Bcl2 mRNA expression in the invasive group ( r =-0.42,P =0.028 ).Conclusions Beclinl expression is decreased in invasive pituitary adenomas.The invasiveness of pituitary adenoma is closely related to the high expression of Bcl2 protein and mRNA,and the low expression of Beclin1 protein and mRNA.The inhibition of the autophagy may lead to the enhancement of the invasiveness of pituitary adenomas and that inhibition may come from the interaction of Beclin1 and Bcl2.
8.2019 Novel Coronavirus (COVID-19) Pneumonia: Serial Computed Tomography Findings
Jiangping WEI ; Huaxiang XU ; Jingliang XIONG ; Qinglin SHEN ; Bing FAN ; Chenglong YE ; Wentao DONG ; Fangfang HU
Korean Journal of Radiology 2020;21(4):494-497
From December 2019, Coronavirus disease 2019 (COVID-19) pneumonia (formerly known as the 2019 novel Coronavirus [2019-nCoV]) broke out in Wuhan, China. In this study, we present serial CT findings in a 40-year-old female patient with COVID-19 pneumonia who presented with the symptoms of fever, chest tightness, and fatigue. She was diagnosed with COVID-19 infection confirmed by real-time reverse-transcriptase-polymerase chain reaction. CT showed rapidly progressing peripheral consolidations and ground-glass opacities in both lungs. After treatment, the lesions were shown to be almost absorbed leaving the fibrous lesions.
9.2019 Novel Coronavirus (COVID-19) Pneumonia: Serial Computed Tomography Findings
Jiangping WEI ; Huaxiang XU ; Jingliang XIONG ; Qinglin SHEN ; Bing FAN ; Chenglong YE ; Wentao DONG ; Fangfang HU
Korean Journal of Radiology 2020;21(4):501-504
From December 2019, Coronavirus disease 2019 (COVID-19) pneumonia (formerly known as the 2019 novel Coronavirus [2019-nCoV]) broke out in Wuhan, China. In this study, we present serial CT findings in a 40-year-old female patient with COVID-19 pneumonia who presented with the symptoms of fever, chest tightness, and fatigue. She was diagnosed with COVID-19 infection confirmed by real-time reverse-transcriptase-polymerase chain reaction. CT showed rapidly progressing peripheral consolidations and ground-glass opacities in both lungs. After treatment, the lesions were shown to be almost absorbed leaving the fibrous lesions.
Adult
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China
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Coronavirus
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Fatigue
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Female
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Fever
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Humans
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Lung
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Pneumonia
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Thorax
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Tomography, X-Ray Computed
10.Reproducibility analysis of quantitative susceptibility mapping of cerebral subcortical nuclei in healthy adults.
Huan XU ; Qinglin MENG ; Wenping FAN ; Xue WANG ; Mengqi LIU ; Zhiye CHEN
Journal of Southern Medical University 2020;40(12):1810-1815
OBJECTIVE:
To investigate the intra- and inter-scanner reproducibility of quantitative susceptibility mapping (QSM) of cerebral subcortical nuclei in healthy adults.
METHODS:
QSM was performed in 21 healthy adults on two different 3.0T MR scanners, and the region of interest (ROI) method was used to measure the magnetic susceptibility value of the left subcortical nuclei (the head of the caudate, putamen, globus pallidus, thalamus, substantia nigra and red nucleus). The intraclass correlation coefficient (ICC) and Bland-Altman method were used to evaluate the inter-scanner and intra-scanner reliability.
RESULTS:
The ICCs of the susceptibility value ranged from 0.90 to 0.99 for all the subcortical gray nuclei except for the head of the caudate nucleus measured on the same MR scanner by the same observer. Bland-Altman analysis revealed that the points with susceptibility differences for all the subcortical gray nuclei except for substantia nigra located in the 95% CI of limits of agreement for the same MR scanner. The ICCs of the susceptibility value for the inter-scanner was 0.49 (0.08-0.75) for the head of the caudate nuleus, 0.80 (0.57-0.91) for the putamen, 0.77 (0.51-0.90) for the globus pallidus, 0.78 (0.54-0.91) for the thalamus, 0.80 (0.56-0.91) for the substantia nigra and 0.93 (0.83-0.97) for the red nucleus. The points with susceptibility difference (95.2%, 20/21) located in the 95% CI of limits of agreement for the putamen and the thalamus measured on two different MR scanners.
CONCLUSIONS
The intra-scanner reproducibility of QSM of the subcortical gray nuclei is superior to the inter-scanner reproducibility in healthy adults.
Adult
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Brain/diagnostic imaging*
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Gray Matter
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Humans
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Iron
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Magnetic Resonance Imaging
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Reproducibility of Results
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Substantia Nigra/diagnostic imaging*