Objective To quantify the effect of 650 nm semiconductor laser irradiation on cutaneous wound healing in rabbits. Methods Model wounds were induced in 21 adult male rabbits. They were then divided randomly into a control group, a 5 mW irradiation group and a 10 mW irradiation group. Biometry and light microscopy were used to investigate the effect of low intensity laser therapy (LILT) on cutaneous wound healing. Results Statistically significant differences were observed among the groups in terms of wound shrinkage. The differences between the irradiated and control groups were significant at the 5th, 9th and 13th day. However, any differences between the S mW and 10 mW groups were not significant. Conclusions LILT at 650 nm at either 5 mW or 10 mW and 1 J/cm2 accelerates cutaneous wound healing in rabbits. But irradiation at 10 mW is no more effective than at 5 mW.

Objective To assess the role of muhi-detector row CT (MDCT) in preoperative evaluation of living renal donors. Methods The data of 104 potential donors who underwent MDCT were retrospectively analyzed. All the candidates underwent 64-MDCT examination. First,unenhanced scans were performed on the kidneys. After administration of Ⅳ contrast medium,enhanced CT images of the arterial phase, venous phase, and excretory phase were obtained. The enhanced scan scope was from the top of diaphragmatic muscle to pubic symphysis. The scanning data obtained was post-processed for reconstructed images. The anatomy and variations displayed in MDCT images on kidneys, ureters, arteries and veins were recorded. The findings in surgery constituted the standard of reference for imaging findings, and the recorded results from images were compared with the findings in surgery to assess the role of MDCT in evaluation of potential donors. Results MDCT examination was successfully performed on 104 candidates. Anomalies of kidneys and ureters were found in 8 donors before surgery. The prevalence of accessory arteries and early branching in image was 27. 2 % (28/103) and 12. 6 % (13/103) respectively. There were 3 candidates with double veins and 3 with retroaortie left renal vein found in preoperative assessment. Ninety-three candidates underwent successful donor nephreetomy. The anomalies and variations of kidneys and ureters in images were all confirmed surgically. The detection rate of the accessory renal artery (ARA) was 80 %. The ARAs measuring > 1 mm in diameter and early branching were all detected by MDCT.The findings of veins found in performed sides coincided with those of MDCT images. Conclusion MDCT can accurately assess the anatomic information and variations of the donors' kidneys, and facilitate triaging donors and planning operation proposal

Objective To explore the expression of anti-MICA antibodies and evaluate its influence on acute rejection and renal function in early period after renal transplantation. Methods A total of 29 sensitized subjects (PRA＞20 %) were enrolled in this study. All the patients underwent protein A immunoabsorption treatment and the expression of anti-MICA antibodies was detected before and after treatment. Triple immunosuppressive regimen consisting of tacrolimus, mycophenolate mofetil (MMF) and steroid was given to prevent graft rejection. The correlation between the expression of anti-MICA antibodies and acute rejection or serum creatinine (SCr) level was analyzed.Results The expression of anti-MICA antibodies was detected in 8 candidates (27. 6 % ,8/29) ,and 6 kinds of anti-MICA antibodies simultaneously expressed were found in one individual, 3 kinds in one case,and sole kind in 6 patients. There was no significant difference in acute rejection rate between positive anti-MICA antibodies group and negative group [37.5 % (3/8) vs 38. 1% (8/21), P＞0.05). The positive expression rate of anti-MICA antibodies in the recipients with PRA ≥40% was higher than that in those with PRA ＜40% [43. 8 % (7/16) vs 7. 7 % (1/13),P＜0.05]. The SCr level in patients positive for anti-MICA antibodies was markedly higher than that in those negative anti-MICA antibodies at the 1st week postoperatively ( 135.4 ± 21.4 vs 108. 6 -+ 31.6 μmol/L, P＜0.05). The SCr level in the patients with positive anti-MICA antibodies, however, was reduced to the normal range at the 2nd week after surgery (P＞0.05). The levels of anti-MICA antibodies were continuously decreased in the candidates undergoing protein A irnmunoadsorption treatment. Conclusion Higher expression of anti-MICA antibodies exists in sensitized recipients and possesses an influence on the recovery of renal function in early postoperative period. Protein A immunoadsorption can eliminate anti-MICA antibodies effectively in sensitized recipients.

The integrity and transparency of cornea plays a key role in vision. Limbal Stem Cells (LSCs) are precursors of cornea, which are responsible for self-renewal and replenishing corneal epithelium. Though it is successful to cell replacement therapy for impairing ocular surface by Limbal Stem Cell Transplantation (LSCT), the mechanism of renew is unclear after LSCT. To real time follow-up the migration and differentiation of corneal transplanted epithelial cells after transplanting, we transfected venus (a fluorescent protein gene) into goat LSCs, selected with G418 and established a stable transfected cell line, named GLSC-V. These cells showed green fluorescence, and which could maintain for at least 3 months. GLSC-V also were positive for anti-P63 and anti-Integrinbeta1 antibody by immunofluorescent staining. We founded neither GLSC-V nor GLSCs expressed keratin3 (k3) and keratinl2 (k12). However, GLSC-V had higher levels in expression of p63, pcna and venus compared with GLSCs. Further, we cultivated the cells on denude amniotic membrane to construct tissue engineered fluorescent corneal epithelial sheets. Histology and HE staining showed that the constructed fluorescent corneal epithelial sheets consisted of 5-6 layers of epithelium. Only the lowest basal cells of fluorescent corneal epithelial sheets expressed P63 analyzed by immunofluorescence, but not superficial epithelial cells. These results showed that our constructed fluorescent corneal epithelial sheets were similar to the normal corneal epithelium in structure and morphology. This demonstrated that they could be transplanted for patents with corneal impair, also may provide a foundation for the study on the mechanisms of corneal epithelial cell regeneration after LSCT.
Animals ; Cell Culture Techniques ; methods ; Cell Line ; cytology ; Epithelium, Corneal ; cytology ; metabolism ; Fluorescent Antibody Technique, Indirect ; Goats ; Limbus Corneae ; cytology ; Stem Cell Transplantation ; Stem Cells ; cytology

Chemically defined medium is widely used for culturing mouse embryonic stem cells (mESCs), in which N2B27 works as a substitution for serum, and GSK3β and MEK inhibitors (2i) help to promote ground-state pluripotency. However, recent studies suggested that MEKi might cause irreversible defects that compromise the developmental potential of mESCs. Here, we demonstrated the deficient bone morphogenetic protein (BMP) signal in the chemically defined condition is one of the main causes for the impaired pluripotency. Mechanistically, activating the BMP signal pathway by BMP4 could safeguard the chromosomal integrity and proliferation capacity of mESCs through regulating downstream targets Ube2s and Chmp4b. More importantly, BMP4 promotes a distinct in vivo developmental potential and a long-term pluripotency preservation. Besides, the pluripotent improvements driven by BMP4 are superior to those by attenuating MEK suppression. Taken together, our study shows appropriate activation of BMP signal is essential for regulating functional pluripotency and reveals that BMP4 should be applied in the serum-free culture system.
Animals ; Bone Morphogenetic Protein 4/metabolism* ; Cell Differentiation ; Chromosomal Instability ; Endosomal Sorting Complexes Required for Transport ; Mice ; Mitogen-Activated Protein Kinase Kinases/metabolism* ; Mouse Embryonic Stem Cells/cytology* ; Pluripotent Stem Cells/cytology* ; Signal Transduction ; Ubiquitin-Conjugating Enzymes

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development