1.Establishment of leukocyte-associated immunoglobulin like-receptor 2 (CD306) eukaryotic expression vectors and purification and identification of fusion protein
Chunyan WANG ; Zhenhua KANG ; Xin XIE ; Yan LI ; Boquan JIN
Chinese Journal of Tissue Engineering Research 2009;13(50):9928-9932
BACKGROUND: Biological function of leukocyte-associated immunoglobulin like-receptor 1 (LAIR 1) has clearly researched in China and abroad, but the in vivo biological function of LAIR is poorly understood. OBJECTIVE: To establish LAIR-2 (CD306) eukaryotic expression vectors and to purify and identify the fusion protein. DESIGN, TIME AND SETTING: A single sample observation experiment was performed at the Fourth Military Medical University of Chinese PLA between June 2007 and June 2008.MATERIALS: plg/3c vector was offered by Oxford University, pcDNA3.1 vector was provided by Meyaard doctor. Chinese hamster ovary (CHO) cell lines were preserved by the Department of Immunology, Fourth Military Medical University of Chinese PLA.METHODS: Two eukaryotic expression vectors plg/3c-LAIR-2 and pcDNA3.1-LAIR-2 were constructed and were transfected into CHO cells. The binding activities of LAIR-2 fusion protein to LAIR-2 mAbs were identified by Western blot, immunocytochemistry and flow cytometry assay.MAIN OUTCOME MEASURES: The construction of stably transfected cell lines, and the purification and identification of fusion protein. The activity of LAIR 2 protein combined to corresponding monoclonal antibody.RESULTS: Eukaryotic expression vectors were constructed and trasnsfected into CHO cells successfully. Two cells lines CHO/LAIR-2-Fc and CHO/LAIR-2 that steadily expressed LAIR-2-Fc fusion protein and LAIR-2 protein were established. Western blot assay showed that LAIR-2 protein could bind specially to LAIR-2 mAb 1A7, 3H12 and 4A9. Immunocytochemistry and flow cytometry assay demonstrated that 3H12 and 4A9 could bind to LAIR-2 expressed in the transfected CHO cells. CONCLUSION: Two ceils lines CHO/LAIR-2-Fc and CHO/LAIR-2 were successfully constructed, which can transfected to CHO cells. The eukaryotic expressed LAIR-2 protein has good binding activity to LAIR-2 mAbs.
2.Long-term outcome of female orthotopic ileal neobladder
Rongxiang ZHOU ; Zongliang ZHANG ; Monong LI ; Haiyan QI ; Yanlun ZHANG ; Rongchen YANG ; Shuai WU ; Yan ZHANG ; Gang ZHANG ; Boquan YAN
Chinese Journal of Urology 2008;29(12):818-821
Objective To evaluate the clinical application of female orthotopic ileal neobladder.Methods Modified radical cysteetomy plus orthotopic ileal neobladder was performed on 19 female pa-tients with bladder cancer from June 1999 to January 2008.The mean age of the patients was 52(45-66) years,mean course of disease was 4.4 months (16 days-1.9 years).Of all the patients,there were 10 cases with grade 1,7 cases with grade 2 and 2 cases grade 3.According to the UICC stage system,5 patients were T1 stage,12 T2 and 2 T3a.All the patients received modified radical cystecto-my without resection of uterus and anterior vagina,meanwhile the nerves around urethra were protec-ted.0.8-1.2 cm proximal end of the urethra was excised and 30 cm distal ileum was used for the re-construction of the neobladder.Results Sixteen cases were followed up for 6-102 months,mean 71 months.Fifteen patients survived without disease recurrence,1 patient died of myocardial infarc-tion 17 months postoperation.The daytime and night continent rate was 100$,93% at 9 months postoperative.The average voiding volume of the 15 patients was 519.0 ml.The average residual vol-ume was 29.2 ml,and Qmax was 18.6 ml/s.The average filling and voiding pressure was 16.7 cm H2O and 53.0 cm H2O.Intravenous urography showed slight hydronephrosis in 1 case.Conclusion Female orthotopic ileal neobladder could be a good choice because of the continence,fewer complica-tions,lower pressure and enough bladder capacity.
3.A Case of Congenital Megalourethra.
Boquan YAN ; Jin Ik PARK ; Ki Hyun YOON ; Gyung Tak SUNG ; Jin Han YOON
Korean Journal of Urology 1995;36(2):218-221
Megalourethra is a rare congenital abnormaly characterized by severe dilatation of urethra, the disorder at first reported by Nesbitt in 1955. The scaphoid type is due to poor development of the corpus spongiosum, whereas in the more severe fusiform variety the corpora cavernosa also are affected. We present a case of congenital scaphoid megalourethra associated with acute renal failure, sepsis and grade IV bilateral vesicoureteral reflux. Initial management of this patient was vesicocutaneostomy and urethrocutaneostomy followed later by urethral reconstruction.
Acute Kidney Injury
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Dilatation
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Humans
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Sepsis
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Urethra
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Vesico-Ureteral Reflux