Abstract:Objective To explore the use of bio?orthogonal click chemistry indocyanine green (ICG) labeling of adipose tissue?derived mesenchymal stem cells (ADSC) and in vivo cell tracking. Methods ADSCs were isolated and cultured, and then incubated with N?azidoacetylmannosamine?tetraacylated ( Ac4ManNAz) and Dibenzocyclooctyne?indocyanine green (DBCO?ICG) using bio?orthogonal click chemistry, and their cell viability was evaluated. ICG labeling was confirmed by laser confocal microscopy imaging. After ICG labeling, in vivo ADSC tracking was performed by near?infrared Ⅱ fluorescence imaging in acute liver injured mice. Liver tissue sections were also collected to analyze ADSC homing. Moreover, the therapeutic effects of ICG?labeled ADSCs on serum levels of alanine aminotransferase (ALT) and aspartate transaminase (AST) and on pathological changes were also evaluated. Results ICG labeling of ADSCs could be achieved by bio?orthogonal click chemistry. Notably, ADSC viability and their therapeutic effects on acute liver injury, including serum ALT and AST and hepatic morphology, were not affected by this method. Near?infrared Ⅱ fluorescence imaging revealed the hepatic accumulation and homing of transplanted ADSCs in vivo and ex vivo. Conclusions Bio?orthogonal click chemistry may provide a promising new strategy for ADSC labeling and in vivo cell tracking.

Abstract:We aimed to characterize the blood reflux, blood compatibility, and biocompatibility of a blood pump after deactivating the pump in an animal in order to facilitate further studies and clinical application. Methods A left- ventricular assist device was implanted into a healthy sheep via a left thoracic approach. After 6 weeks of normal operation, the pump was stopped, as was warfarin administration. The physiological status of the animal was then monitored, and blood biochemical and coagulation testing was performed before the procedure and between 1 and 12 weeks after the pump was deactivated. Ultrasonographic examination was performed 16 weeks after the pump was deactivated. Results The left-sided cardiac assistance model was established successfully, and 6 weeks after deactivation of the pump the sheep displayed normal behavior and normal physiology, with no significant changes in the test result . Ultrasonography showed that the artificial blood vessel was not blocked and there was reverse blood flow 16 weeks after the blood pump was stopped. Conclusions Deactivation of a blood pump was not associated with effects on blood compatibility, physiological state, or organ function in a sheep. The artificial blood vessel was not obstructed, no thrombosis was found, and left-sided cardiac function was not significantly affected. Thus, this blood pump has good compatibility, and the study has provided a reference for the clinical treatment of unexpected pump failure. Further studies should assess the impact of low-dose anticoagulant therapy and repeated deactivation on physiology and cardiac function.

Abstract:Objective　To establish an in vivo model to assess the risk of particle embolization of drug-coated balloons in animals, and to explore the relationship between the diameter of occluded vessels and the particle size of the coating. Methods　The distal end of abdominal aorta was blocked with a bare balloon, and then a drug coated balloon catheter was delivered into the abdominal aorta of the rabbit and dilated at the proximal end to observe whether the kidneys had tissue necrosis caused by typical vascular embolization. Results　Abnormal lesions of the kidneys are mainly characterized by volume shrinkage, pits, and roughness. Histological examination showed glomerular congestion, loss of cell nuclei and karyolysis, fibrosis, and inflammatory cell infiltration. The pathological process of renal necrosis occurred from the cortex to the medulla, the extent of necrosis was related to the dose, and there was no improvement by self-healing over time. Conclusions　The rabbit model we designed is sensitive and can be used to assess the risk of embolization from drug-coated balloons in the body.

Abstract:Objective　The fluorescent dye Nile red (NR) is commonly used in quantitative analysis of lipids in bacteria, fungi, and microalgae. In this study, NR was encapsulated in a nanoemulsion to investigate the tumor targeting and tissue distribution of the nanoemulsion in BALB/ c nude mice bearing H1688 tumors. Methods 　To evaluate the prospects of NR as a bioimaging marker for nanoemulsions, its cytotoxicity was investigated in H1688 cells with a CCK-8 assay. An NR suspension (NRS) and NR nanoemulsion NRNE(O) were given by gavage. The dynamic fluorescence intensity distribution of NR in mice was observed with an in vivo fluorescence imaging system. Results　The H1688 cells exhibited over 85% viability after incubation with a range of NRNE(O) concentrations for 24 h. The results demonstrated that NRNE(O) absorption in mice and the amount accumulated in tumor tissue were both higher than in the NRS group.Conclusions　NRNE(O) has a low cytotoxicity and good biocompatibility. NR in a nanoemulsion combined with an in vivo fluorescence imaging system can reveal the dynamic distribution of the nanoemulsion in mice vividly and reliably in real time. The results of this study indicate that NR has significant application potential as a tracer for the bioimaging of nanoparticles in vivo.

Abstract:Objective To investigate the differential expression of integrin alpha v beta 3(αv,β3) on bovine endometrial epithelial cells before and after co-culture with bovine blastocysts, and to explore whether this specific signal might be applied as a new marker for identifying the bovine uterine receptivity. Methods The in vivo bovine embryos were co-cultured with their endometrial epithelial cells for 24 h, then, RT-PCR was used to detect the differential expression of αv,β3 among those groups. Result The results showed that αv,β3 can be expressed on bovine endometrial epithelial cells both before and after co-culture with their in vivo embryos. There were no significant differences of expression of αv,β3 between Group I (only bovine blastocyst) and the control one (P > 0.05), but there were significant differences of αv,β3 among Group Ⅱ (the hatched bovine blastocyst), and Group I, the control one (P< 0.05). Conclusions The in vivo hatched bovine blastocysts are more suitable for induction of intergrin αv,β3 in bovine endometrial epithelial cells after their co-culture than that of co-cultured with early stage blastocyst. Integrin αv,β3 might be applied as a new molecular marker for detecting bovine uterine receptive status of the bovine endometrium.

Abstract:ObjectiveTo study the early dynamics of lesion formation after vascular injury in LDLR-/- mice by in vivo fluorescence microscopy. MethodsOne hundred and twelve male LDLR-/- mice (18-22 weeks of age) were randomly divided into 14 groups (8 mice in each group). The mice were subjected to bone marrow cell transplantation from GFP+/LDLR-/- mice. After 4 weeks, a polyethylene cuff was implanted around the right femoral artery to induce vascular injury. The lesion formation was observed on the injured femoral artery from 1 to 14 days after vascular injury.ResultsAt the 1st day after cuff placement, a markedly large number of GFP positive cells were clearly observed in high-speed blood flow. At the 3rd day after the vascular injury, GFP positive cells accumulated in the inner vascular wall and formed punctate lesions. At the 6th day after the vascular injury, GFP positive cells in the inner vascular wall formed irregular flakes, and a small amount of the adventitia tissue proliferated, corresponding to the inner lesion area. At the 9th day after the vascular injury, the adventitia tissue was obviously proliferated and a large number of GFP positive cells embeded. The vasa vasorum were clearly seen running in the adventitial layer. At the 14th day after the vascular injury, the adventitia tissue proliferated considerably with largely GFP positive cells and could not be observed in the lesion through adventitia. After stripping the outer tissue of vessel wall, the fluorescent lesion formed by GFP positive cells was clearly lining in the intima area.ConclusionsThe results of our study demonstrate that the vascular remodeling lesion in the early stage in LDLR-/- mice has a trend of “inside out”. The dynamics of lesions has a close correlation with bone marrow-derived cells that circulating in the blood flow. There is distinct influence of the vascular lesions on extravascular fibrosis. 

Abstract:Objective In order to study the effect of the genotype of different strains on mouse embryo cryopreservation in the glycerol vitrification solution.Methods The 6 5mol/L glycerol and 6% (w/v) BSA were used as the vitrification solution for the cryopreservation of morulae and blastocysts embryos of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse.Results and Conclusion The rates of in vitro development of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse embryos to expanded blastocysts ranged from 31 3% to 88% after vitrification.The rates of CBA,NOD,C 57 BL/6J,ICR and CD1 mouse embryos to normal fetuses was 21%,23 5%,11%,38%,35 5%.The rates of in vitro development of CBA,NOD,C 57 BL/6J6,ICR and CD1 mouse morulae and early blastocysts were significantly higher than expanded blastocystst.The in vitro development and in vivo development were significantly influenced by inbred strain,closed colony and different stages of mouse embryo.The result of mouse embryo cryopreservation in the vitrification solution may be connected with the different genotype of the different strains and different stages of mouse embryo.

Abstract:It has estimated that MID100(100% monkey infected dose) of SIVmac251 was 32 TCID andSIVmac239 was more than 320 TCID. Three monkeys inoculated with SIVmac251, two monkeysSIVmac239 and seven monkeys SIVmac251 2 MID100 were infected pefectly. All infected monkeysappeared generalized lymphopathy, plasma viremia and antibody response regularly. The pathological examination of lymph nodes and spleens in seven Rhesus and two cynomolgus monkeysshowed infectious changes. It is quite obvious that the symptoms an sign, Pathological histologicalchanges, and viremia and antibody response of infected monkeys were caused by SIVmac251 orSIVmac239 strain. So above strains could be used to evaluate the effect of anti - AIDS drugs inmonkey test.