1.Investigation and analysis of clinical application of washed red blood cells
Rongxian LIAO ; Guoying FU ; Shaoyun WU ; Xuemei XU ; Hao LV
Chinese Journal of Blood Transfusion 2017;30(7):760-761
Objective To investigate and analyze the clinical application of washed red blood cells in Quzhou in 2014-2015 years,and to provide evidence for the rational and effective use of washed red blood cells in clinical practice.Methods the amount of washed red blood cells,the distribution of diseases and the serological detection before transfusion were analyzed retrospectively in 8 hospitals of grade two and over in Quzhou during the past 2014-2015 years.Results the amount of washed red blood cells in 2015 increased by 39.07% compared with 2014,an increase of 24.4 times the amount of red blood cells increase,increase mainly in 3 hospitals;Diseases of the blood system,malignant tumors and chronic kidney disease is the main diagnosis of transfusion cases,accounted for 63.49%,19.05%,12.70%.There were different standards for the development of serological testing items before transfusion Conclusion the advantages of washed red blood ceils gradually recognized by clinicians,but also don't rely on experience,hospitals should pay attention to the comprehensive evaluation of clinical blood transfusion and standardize the serological detection of blood transfusion department,which is the key to improve the cost performance of this component.
2.Imbalance of MMP-9/TIMP-1 during the cutaneous "underlying disorder" in diabetic rats
Li YAN ; Ping ZHU ; Lihong CHEN ; Chuan YANG ; Guojuan LAO ; Jing DU ; Shaoyun HAO ; Hua CHENG
Chinese Journal of Endocrinology and Metabolism 2008;24(5):533-536
Objective To investigate the change of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the skin of diabetic rats, and to explore the potential role of MMP-9/TIMP-1. Methods Diabetic rats were induced with streptozotocin (STZ). Then all rats were maintained for 6 weeks. Routine pathological examination and immnnohistochemistry were made to reveal the histological and cytological appearances. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin. Results Six weeks after STZ treatment, examination of HE-stained skin sections from normal and diabetic animals revealed that the epidermis and dermis layers were thinner in diabetic rats than those in control rats. The skin of diabetic rats showed features of atrophy such as disorganization of connective tissue fiber bundles and enlarged space between collagen fiber bundles. In contrast, thick bundles of connective tissue were observed in the dermis of normal rat skin. In normal skin, cells had a bipolar, spindle-shaped appearance in the thick collagen bundles, while in the skin of diabetic animals the interstitial cells had a rounded, shrunken and crenated appearance. The relative values of expression of MMP-9 in diabetic group were higher than those in normal group with significant difference, however, the relative values of expression of TIMP-I in diabetic group were lower than those in control group. Conclusion The changes in cutaneous histology and cytology appear earlier than skin wound. These "underlying disorders" may be associated with the imbalance of MMP-9/TIMP-1.
3.Construction of tissue-engineered skin using basic fibroblast growth factor gene transfected bone marrow mesenchymal stem cells combined with small intestinal submucosa
Shaoyun WANG ; Hao YANG ; Xiaosong ZHU ; Qing LI ; Shihe LI ; Jian DONG ; Di WU
Chinese Journal of Tissue Engineering Research 2010;14(7):1174-1178
BACKGROUND: Studies demonstrated that small intestinal submucosa (SIS) had no immunogenicity, which can not lead to rejection following transplantation, thus, this is an ideal skin substitutes for natural skin.OBJECTIVE: Basic fibroblast growth factor (bFGF) gene was transfected into bone marrow mesenchymal stem cells (BMSCs),and combined the cells with SIS to construct tissue-engineered skin.METHODS: BMSCs were obtained from Japanese big-earad rabbits, and in vitro cultured. Then the subculturad BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry. In addition, the result of transfecting BMSCs with pCDNA-bFGF vector was measured by Western blot, and the structure of tissue-engineered skin was observed.RESULTS AND CONCLUSION: After passaged, BMSCs were grown quickly with Iong-fusiform shape. The cells were positive expressed CD90 and CD44, but negative expressed CD45. bFGF had been transfected into BMSCs, and stable expressed. The transfected BMSCs grew well in SIS. By this method, tissue-engineered skin can be constructed in vitro.
4.Transfection of basic fibroblast growth factor gene into rabbit bone marrow mesenchymal stem cells
Shaoyun WANG ; Di WU ; Xiaosong ZHU ; Hao YANG ; Hongbin ZHAO ; Jian DONG ; Shihe LI
Chinese Journal of Tissue Engineering Research 2009;13(49):9687-9691
BACKGROUND: Basic fibroblast growth factor (bFGF) has significant promotion effects on repair in trauma, but local application cannot play a role for a long time.OBJECTIVE: To observe expression of bFGF gene in rabbit bone marrow mesenchymai stem cells (BMSCs) following transfection.DESIGN, TIME AND SETTING: The cytogene in vitro observation was performed at the College of Life Science, Yunnan University from March to August 2009.MATERIALS: One Japan flap-eared rabbit was purchased from the Department of Animal, Kunming Medical College. pCDNA3.1plasmid (Invitrogen, USA) was used in this study.METHODS: Bone marrow was extracted from rabbit anterior superior iliac spine. BMSCs were harvested by the adherent method.Cells were digested and subcultured when 80% confluent. According to GeneBank bFGF cDNA sequence, gene was designed and synthesized. Following electropherosis, the gel was retrieved using xhol I, BamH I enzyme digestion. Restriction enzyme was used to perform enzyme digestion, electropherosis and gene recovery in PcDNA Vector plasmid. bFGF DNA was connected with PcDNA Vector plasmid. PcDNA-bFGF eukaryotic expression vector was constructed. Recombinant was transfected into rabbit BMSCs using liposome infection protocol, and stable transfected line was screened.MAIN OUTCOME MEASURES: BMSC surface antigen expression was measured. Western blot was utilized to determine the expression of target protein.RESULTS: Results of flow cytometry showed that cultured cells were positive for CD90 and CD44, but negative for CD45.Results of immunohistochemistry demonstrated that vessel-derived BMSCs were negative for CD34, but positive for CD44. In cell disruption liquid of bFGF-transfected BMSCs, a significant positive zone of hybridization was visible at M, 23 000. However, no positive band was found in protein from pCDNA3.1(-) blank vector-transfected BMSCs.CONCLUSION: The bFGF gene was successfully transfected into BMSCs, and this target gene can stably express.