Angelicin improves osteoporosis in ovariectomized rats by reducing ROS production in osteoclasts through regulation of the KAT6A/Nrf2 signalling pathway.

BACKGROUND: Angelicin, which is found in Psoralea, can help prevent osteoporosis by stopping osteoclast formation, although the precise mechanism remains unclear. METHODS: We evaluated the effect of angelicin on the oxidative stress level of osteoclasts using ovariectomized osteoporosis model rats and RAW264.7 cells. Changes in the bone mass of the femur were investigated using H&E staining and micro-CT. ROS content was investigated by DHE fluorescence labelling. Osteoclast-related genes and proteins were examined for expression using Western blotting, immunohistochemistry, tartrate-resistant acid phosphatase staining, and real-time quantitative PCR. The influence of angelicin on osteoclast development was also evaluated using the MTT assay, double luciferin assay, chromatin immunoprecipitation, immunoprecipitation and KAT6A siRNA transfection. RESULTS: Rats treated with angelicin had considerably higher bone mineral density and fewer osteoclasts. Angelicin prevented RAW264.7 cells from differentiating into osteoclasts in vitro when stimulated by RANKL. Experiments revealed reduced ROS levels and significantly upregulated intracellular KAT6A, HO-1, and Nrf2 following angelicin treatment. The expression of genes unique to osteoclasts, such as MMP9 and NFATc1, was also downregulated. Finally, KAT6A siRNA transfection increased intracellular ROS levels while decreasing KAT6A, Nrf2, and HO-1 protein expression in osteoclasts. However, in the absence of KAT6A siRNA transfection, angelicin greatly counteracted this effect in osteoclasts. CONCLUSIONS: Angelicin increased the expression of KAT6A. This enhanced KAT6A expression helps to activate the Nrf2/HO-1 antioxidant stress system and decrease ROS levels in osteoclasts, thus inhibiting oxidative stress levels and osteoclast formation.

The characteristics of postprandial glycemic response patterns to white rice and glucose in healthy adults: Identifying subgroups by clustering analysis.

Objectives: Large interpersonal variability in postprandial glycemic response (PGR) to white rice has been reported, and differences in the PGR patterns during the oral glucose tolerance test (OGTT) have been documented. However, there is scant study on the PGR patterns of white rice. We examined the typical PGR patterns of white rice and glucose and the association between them. Materials and methods: We analyzed the data of 3-h PGRs to white rice (WR) and glucose (G) of 114 normoglycemic female subjects of similar age, weight status, and same ethnic group. Diverse glycemic parameters, based on the discrete blood glucose values, were calculated over 120 and 180 min. K-means clustering based on glycemic parameters calculated over 180 min was applied to identify subgroups and representative PGR patterns. Principal factor analysis based on the parameters used in the cluster analysis was applied to characterize PGR patterns. Simple correspondence analysis was performed on the clustering categories of WR and G. Results: More distinct differences were found in glycemic parameters calculated over 180 min compared with that calculated over 120 min, especially in the negative area under the curve and Nadir. We identified four distinct PGR patterns to WR (WR1, WR2, WR3, and WR4) and G (G1, G2, G3, and G4), respectively. There were significant differences among the patterns regard to postprandial hyperglycemia, hypoglycemic, and glycemic variability. The WR1 clusters had significantly lower glycemic index (59 ± 19), while no difference was found among the glycemic index based on the other three clusters. Each given G subgroup presented multiple patterns of PGR to WR, especially in the largest G subgroup (G1), and in subgroup with the greatest glycemic variability (G3). Conclusion: Multiple subgroups could be classified based on the PGR patterns to white rice and glucose even in seemingly homogeneous subjects. Extending the monitoring time to 180 min was conducive to more effective discrimination of PGR patterns. It may not be reliable to extrapolate the patterns of PGR to rice from that to glucose, suggesting a need of combining OGTT and meal tolerance test for individualized glycemic management.

Anatomical study of anterior supramalleolar artery and its potential application to design a bi-foliate fasciocutaneous flap.

PURPOSE: A further understanding of the anterior supramalleolar artery (ASMA) and its potential applications in reconstructive surgery. MATERIALS AND METHODS: A total of 24 fresh lower limbs from fresh cadavers were injected with red latex for dissection. The type of origin, course, diameter of the pedicle, and the distance between the origin of the ASMA from the anterior tibial artery to the extensor retinaculum (O-R) were recorded. Bi-foliate fasciocutaneous flaps were harvested using the branches of the ASMA. RESULTS: We found four types of origin of the ASMA, and we have accordingly classified them into four types. 10 of them were type A, 7 were type B, 6 were type C and 1 was type D. The mean O-R (origin of ASMA to retinaculum) distance was 2.0 ± 0.8 cm. The diameter of the medial branch (D1), the diameter of the lateral branch (D2), and the diameter of artery stem (D3) (only in type A) were 1.0 ± 0.2 mm, 0.8 ± 0.3 mm, 1.1 ± 0.2 mm, respectively. The mean pedicle length of the lateral flap (L1) and medial flap (L2) were 5.1 ± 1.0 cm and 3.7 ± 0.6 cm, respectively. CONCLUSIONS: The ASMA exists constantly with four different types of origin. Its sizable diameter and lengthy pedicle make it suitable for bi-foliate fasciocutaneous flap transfer.

Is the Resorption of Grafted Fat Reduced in Cell-Assisted Lipotransfer for Breast Augmentation?

Cell-assisted lipotransfer (CAL) is a cotransplantation of adipose tissue and stromal vascular fraction (SVF) including adipose-derived stem cells. But although CAL can get satisfactory outcomes in breast augmentation, the resorption of the grafted fat is still unclear. A total of 12 patients received breast augmentation using CAL. All of them completed 6 months of follow-up. In 1 mini-CAL case, 500-mL liposuction fluid was used to harvest the SVF cells. In 11 full-CAL cases, 250-mL aspirated fat was needed apart from 500-mL liposuction fluid. The percentage of adipose-derived stem cells in SVF cells was detected using flow cytometry and their multilineage potential ability was assessed with in vitro induction. The volumes of breasts and pectoral muscle were measured, and radiological image change was analyzed using magnetic resonance imaging before the operation and 3 and 6 months after the operation. Additionally, the subjective evaluation on the cosmetic outcomes was determined by surgeons and patients. Adipose-derived stem cells in SVF cells accounted for 40.27% and 3.34% in full-CAL cases and mini-CAL cases, respectively. Postoperative atrophy occurred within the first 3 months. At the 6 months postoperatively, breast volume is augmented, ranging from 60.71 to 197 mL, with a mean value of 125.35 (45.49) mL. The ultimate resorption of grafted fat at the 6 months postoperatively is 51.84% (16.74%). Newly formed cysts and nodules were detected in 2 cases. No calcification was found in all magnetic resonance images. Only 1 patient was unsatisfied with the cosmetic outcome. Our preliminary study displayed a satisfactory augmented volume with little complications using CAL for breast augmentation. But the resorption at the 6 months postoperatively [51.84% (16.74%)] showed no significant advantage over non-CAL technique (40%-60% reported), which suggested that SVF cells harvested from 250-mL aspirated fat and 500-mL liposuction fluid were insufficient to average 250-mL grafted fat for each breast in this study. More SVF cells are needed to achieve lower resorption.

Trifocal distraction osteogenesis for reconstruction of skull defect.

OBJECTIVE: To apply trifocal distraction osteogenesis in canine model of skull segmental defects and to provide reference for clinical treatment. METHODS: Six labrador dogs were selected in this study and divided into observation group and control group randomly. Each group contained 3 dogs. Skull segmental defects models were established by surgery, and dogs in bservation group received trifocal distraction osteogenesis treatment. Bone density was observed and compared between two groups during treatment. RESULTS: There were no significant difference in bone density between two groups on th 1st day (P>0.05). The bone density of observation group on the 30th day, and 60th day were higher than that of control group (P<0.01). CONCLUSIONS: Trifocal distraction osteogenesis has significant clinical effect, and it would be widely used in clinical treatment.

[Cell-assisted lipotransfer for breast augmentation: a report of 18 patients].

OBJECTIVE: To evaluate the clinical effect of cell-assisted lipotransfer (CAL) for breast augmentation. METHODS: 18 patients accepted breast augmentation using CAL. 10 patients completed 6-month follow-up and were involved in the study. The adipose tissue was harvested from patients' thighs, flanks and lower abdomen with Lipokit. After standing, 250 ml fatty portion and 500 ml fluid portion of suction aspirates were processed according to the procedures reported in reference. Flow-cytometry was used to detect the percentage of adipose-derived stem cells(ADSCs) in distilled stromal vascular fraction (SVF). The differentiation function of cultured cells also was assessed. The breast volume and images were evaluated by using MRI before operation, 3 and 6 months after operation. The breast volume was marked as V0, V3 and V6 respectively. The resorption rate of transplanted adipose tissue for each breast was calculated and marked as R3 and R6. RESULTS: Averagely, the percentage of ADSCs in freshly distilled SVF was 41.67%. The in-vitro cultured cell grew well and could differentiate into fat, bone and cartilage. Statistics showed that V0, V3 and V6 was (416.19 +/- 40.43) ml, (551.72 +/- 59.86) ml and (538.81 +/- 68.35) ml respectively. R3 and R6 was (51.20 +/- 11.96)% and (54.22 +/- 12.73)%. There was significant difference between V3 and V0 (P < 0.05), V6 and V0. However, no significant difference was showed between V3 and V6 or R3 and R6. In addition, no cyst or calcification was seen in all MRI images. CONCLUSIONS: In process of breast augmentation using CAL, the distilled SVF contains 41.67% ADSCs which have adipogenic, osteogenic and chondrogenic function. Within 3-month post-operation, the breast volume decreases obviously but the volume sustains after that. Compared with the preoperative volume, the 6-month postoperative volume is significantly increased and the breasts' contour is improved greatly. This study indicates that CAL is a safe and effective way for breast augmentation.

[Effect of botulinum toxin type A injection on hypertrophic scar in rabbit ear model].

OBJECTIVE: To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. METHODS: The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T (treated with Botox A, n = 48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index (HI). The expression of collagen I and III was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. RESULTS: The HI was significantly lower in group T than in group S (P < 0.01). The expression of collagen I and III, as well as the ratio of I to III, was markedly stronger in group S than in group T (P < 0.01). Compared with group T, more fibroblasts were in G2-M in group S and fewer in G0-G1 (P < 0.05). CONCLUSIONS: Local injection of Botox A can inhibit the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen I and III in hypertrophic scar, as well as the ratio of collagen I to III. It serves as the basis for the treatment of hypertrophic scar with Botox A.

[Effect of botulinum toxin type A on the expression of substance P, calcitonin gene-related peptide, transforming growth factor beta-1 and alpha smooth muscle actin A in wound healing in rats].

OBJECTIVE: To investigate the effect of botulinum toxin type A on the expression of substance P (SP), calcitonin gene-related peptide (CGRP), transforming growth factor beta-1 (TGF-beta1) and alpha smooth muscle actin A (alpha-SMA) in wound healing. METHODS: 60 rats were randomly divided into group C (control) group L (low-dose) and group H (high-dose), with 20 rats in each group. The wound-healing model was established by excision of four full-thickness skin (1 cm x 1 cm, around the injection site) on the back of all SD rats on the 7th day after BTA injection. The wound size was measured and the expression of SP, CGRP, TGF-beta1 and alpha-SMA in wound granulation tissue was assayed by immunohistochemical staining and computerized image analysis before operation, and 3 days, 7 days and 14 days after operation. RESULTS: All the wounds healed 14 days after operation. The wound size in L and H group was not significantly different with that in C group on the 3rd day and 7th day after operation. The positive immuno-staining of SP, CGRP, TGF-beta1 and alpha-SMA in group L and H was significantly weaker than those in C group. Meanwhile, the positive immuno-staining of all above substances in H group was weaker than those in L group significantly. CONCLUSIONS: Botulinum toxin type A can decrease the expression of SP, CGRP, TGF-beta1, and alpha-SMA in wound healing in a dose-dependent manner with no effect on the healing time.

[Effects of antisense oligonucleotides on the expression of focal adhesion kinase gene and collagen synthesis in the cultured human fibroblasts of hypertrophic scar].

OBJECTIVE: To study the role of focal adhesion kinase (FAK) in the pathogenesis of human hypertrophic scar. METHODS: Human hypertrophic scar fibroblasts (HSFB) were isolated from human hypertrophic scar and cultured in vitro. The cells were then divided into 3 groups as AT group (phosphorothioate FAK ASODN was transfected into the HSFB by liposome), LPC group (liposome only), and LC group (control group, without liposome or ASODN). The FAKmRNA index of HSFB was assessed by polymerase chain reaction method (FQ-PCR). The collagen synthesis of HSFB was assessed by 3H-proline incorporation method. RESULTS: The FAK mRNA index of HSFB in AT group 48 hours after transfection was significantly lower than that in LPC and LC groups (0.043 +/- 0.030, 0.124 +/- 0.070, 0.127 +/- 0.0195, P < 0.05). The 3H-proline incorporation rate in AT group was lower than that in LPC and LC groups (257.0 +/- 15.14, 962.2 +/- 300.5, 930.8 +/- 28.97, P < 0.01). CONCLUSION: The expression of FAK gene and collagen synthesis of the cultured HSFB could be inhibited by FAK ASODN, indicating that FAK played a role in the development of excessive fibrosis of human hypertrophic scar.

[The experimental study of mechanism of integrin in hypertrophic scar contracture].

OBJECTIVE: To study the mechanism of integrin in hypertrophic scar. METHODS: Fibroblasts from 10 samples of human hypertrophic scars was cultured, FQ-PCR assay was applied to detect mRNA expression of alpha-SMA in hypertrophic scar fibroblasts after integrin and FAK antibody blocking. RESULTS: mRNA of alpha-SMA in fibroblasts expressed obviously lower after integrin and FAK antibody blocking than that of control groups ( P < 0.05). CONCLUSION: Through accelerating the synthesis of alpha-SMA, integrin and FAK play an important role in contracture of hypertrophic scar.