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Objective To explore the possibility of the inner vertical outer spiral complex tubular urethra scaffold vascularization in repairing long segment of urethral defect.Methods From August 2014 to October 2015,27 clean male New Zealand white rabbits were divided into 3 groups,S1 group was transfected recombinant vascular endothelial growth factor(VEGF) gene lentiviral vector group.S2 group was vascular pedicle transfer tube group.C group was simple stent group.A 3.0 cm inner vertical outer spiral complex scaffold was constructed by using the small intestine acellular matrix (SIS) and polylactic acid copolymer (PLGA) modified by type Ⅰ collagen surface,and adipose-derived mesenchymal stem cells (ADSC) and smooth muscle cells after transformation from New Zealand white rabbits.In S1 group,the seed cells were transfected by recombinant vascular endothelial growth factor (VEGF) gene lentivirus,which express VEGF protein.The complex scaffold was used to repair 3.0 cm rabbit urethral defect In S2 group,the untransfected cells were seeded into the scaffold and embedded in the skin near the groin artery 3 weeks for repairing urethral defect with vascular pedicle transfer tube.In group C,the unseeded scaffold was used to repair the urethral defect alone.Postoperative observation and urethrography were followed 4,8 and 24 weeks after implantation.The HE staining,fluorescence tracing,immunohistochemical and scanning electron microscopy were evaluated at the same phase.Results In S1 group,there were one urinary fistula and one urethral stricture-related death,respectively.The urethra was smooth and patent,histological examination showed active hyperplasia of urethral capillary.In S2 group,there were one urinary fistula and two urethral stricture-related deaths,respectively.The urethral was rough,local thinning or dilated.Fat accumulation and mucosal contraction were found in the urethral submucosal,respctively.In C group,there were one urinary fistula,three hypospadias,and three urethral stricture-related deaths.The thickness of the urethra was uneven and stricture bending.The urethral mucosa was poorly repaired and the scar was narrow.HE and CD31 staining showed that S1 and S2 groups were active in the proliferation of urethral capillaries,and the angiogenesis was abundant.VEGF staining showed that the cytoplasm of endothelial cell layer,smooth muscle layer of vascular wall and the urothelial epithelial cell layer were fully expressed at 24 weeks,especially in epithelial cell layer.CKpan staining showed that the epithelium of S1 and S2 group developed to stratified epithelium,and the morphology of urethra was similar to normal urethra at 24 weeks.The urethral epithelial in C group of grew poor as single-level,irregular arrangement,24 weeks is still a lack of effective stratified epithelium.HE and oα-SMA staining showed that the smooth muscle and actin gradually increased in group S1 and S2,α-SMA staining in group C was scarce and increased at 24 weeks.PLGA was encapsulated by the surrounding tissue and the structure of electrospinning was clear after 4 weeks,absorbed and degraded after 8 weeks and absorbed after 24 weeks.Conclusions The inner vertical outer spiral comnplex tubular urethra scaffold maybe a reasonable method in repairing long segment urethral defects,and the methods of tubular urethra scaffold vascularization by transfected VEGF gene recombinant lentiviral vector and vascular flap deserve more research.
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AIM To establish an LC-MS/MS method for the simultaneous content determination of paeoniflorin,albiflorin,neochlorogenic acid,chlororogenic acid,catechin,epicatechin,ethyl gallate,danshensu,ferulic acid,caffeic acid,gallic acid,protocatechuic acid,protocatechualdehyde and rosmarinic acid in Yangxue Qingnao Granules (Angelicae sinensis Radix,Chuanxiong Rhizoma,Paeoniae Radix Alba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 20 ℃ thermostatic Waters Symmetry Shield RP C18 column (3.9 mm × 150 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 0.4 mL/min in a gradient elution manner.RESULTS Fourteen constituents showed good linear relationships within their own ranges,whose average recoveries were 85.5％-107.0％ with the RSDs of 2.0％-5.0％.CONCLUSION This simple,sensitive,accurate and reproducible method can be used for the quality control of Yangxue Qingnao Granules.
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Objective To explore the effects of microsurgical technique treatment on acknowledge ability ,activities of daily liv-ing ,cortex blood perfusion and neuropeptide levels of brain glioma patients .Methods Microsurgical technique treatment and tradi-tional operation were given to 87 brain glioma patients ,respectively .The acknowledge ability and activities of daily living were measured by HDS ,MMSE and ADL ;the cortex blood perfusion were revealed by PET ,and the neuropeptide levels including argi-nine vasopressin ,oxytocin ,β-endorphin ,somatostatin and neurotensin of patients from two different surgical groups were compared . Results As compared with the traditional operation ,the acknowledge ability and activities of daily living of patients from microsur-gical treatment group were significantly improved(P<0 .05) ,the cortex blood perfusion were statistically increased (P<0 .05) and the levels of five kinds of neuropeptide were all profoundly increased (P< 0 .05) .Conclusion Microsurgical technique treatment could improve the acknowledge ability and activities of daily living of brain glioma patients ,the mechanisms of which could be the increase of cortex blood perfusion and theincrease of neuropeptide levels .