Enhancing Fat Graft Survival via Upregulating Autophagy of Adipocytes.

BACKGROUND: Autophagy is a cellular self-protection mechanism. The upregulation of adipose-derived stem cells' (ADSCs) autophagy can promote fat graft survival. However, the effect of interfering with adipocyte autophagy on graft survival is still unknown. In addition, autophagy is involved in adipocyte dedifferentiation. We investigated the effect of autophagy on adipocyte dedifferentiation and fat graft survival. METHODS: The classic autophagy regulatory drugs rapamycin (100 nM) and 3-methyladenine (3-MA; 10 mM) were used to treat adipocytes, adipocyte dedifferentiation was observed, and their effects on ADSCs were detected. In our experiments, 100 nM rapamycin, 10 mM 3-MA and saline were mixed with human adipose tissue and transplanted into nude mice. At 2, 4, 8 and 12 weeks postoperatively, the grafts were harvested for histological and immunohistochemical analysis. RESULTS: Rapamycin and 3-MA can promote and inhibit adipocyte dedifferentiation by regulating autophagy. Both drugs can inhibit ADSC proliferation, and 10 mM 3-MA can inhibit ADSC adipogenesis. At weeks 8 and 12, the volume retention rate of the rapamycin group (8 weeks, 64.77% ± 6.36%; 12 weeks, 56.13% ± 4.73%) was higher than the control group (8 weeks, 52.62% ± 4.04%; P < 0.05; 12 weeks, 43.17% ± 6.02%; P < 0.05) and the rapamycin group had more viable adipocytes and better vascularization. Compared with the control group, the volume retention rate, viable adipocytes and vascularization of the 3-MA group decreased. CONCLUSIONS: Rapamycin can promote adipocyte dedifferentiation by upregulating autophagy to promote fat graft survival. 3-MA can inhibit graft survival, but its mechanism includes the inhibition of adipocyte dedifferentiation and ADSC proliferation and adipogenesis. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Dietary pattern and precocious puberty risk in Chinese girls: a case-control study.

BACKGROUND: The role of dietary intake on precocious puberty remains unclear. This study aimed to investigate the association between the amount and frequency of dietary intake and the risk of precocious puberty in Chinese girls. METHODS: In this case-control study, we enrolled 185 precocious puberty girls and 185 age-matched controls. Their dietary intake was assessed through a semi-quantitative food frequency questionnaire. Their sociodemographic and lifestyle data were collected. The associations between dietary intake and risk of precocious puberty were assessed by conditional logistic regression models. RESULTS: After multivariate adjustment, consuming a higher amount of red meat was associated with higher precocious puberty risk (OR = 2.74, 95% CI: 1.25-6.02), while a higher frequency of fruit ( P for trend = 0.024) and amount of vegetable intake was associated with a lower risk of precocious puberty (P for trend = 0.002). The high vegetable and protein dietary pattern was significantly negatively associated with precocious puberty (OR = 0.78, 95% CI: 0.63-0.97), whereas the high animal food and fruits dietary pattern was remarkably positively associated with precocious puberty (OR = 1.36, 95% CI: 1.09-1.69), after adjusting for age and body mass index. CONCLUSIONS: High vegetable and protein dietary pattern is a protective factor against precocious puberty, while high animal food and fruits dietary pattern is a risk factor for precocious puberty in Chinese girls. Attentions should be paid to a reasonable intake of red meat, eggs, and fruits in children's daily diet, increase their intake of vegetables, in order to reduce the risk of precocious puberty.

Evaluating the Efficacy of Dedifferentiated Fat Cells (DFATs) vs Adipose-Derived Stem Cells (ASCs) in Enhancing the Viability of Fat Grafts.

BACKGROUND: Enhancing graft fat survival remains a paramount challenge in autologous fat transplantation surgeries. Dedifferentiated fat cells (DFATs) and adipose-derived stem cells (ASCs) represent 2 pivotal cells with potential to improve fat graft survival rates. OBJECTIVES: In this study we aimed to compare the effectiveness of DFATs and ASCs in promoting fat graft survival, emphasizing their adipogenic and angiogenic capabilities. METHODS: Both in vitro and in vivo experiments were conducted. In vitro assessments compared adipogenesis, angiogenesis, osteogenesis, chondrogenesis, cell migration abilities, and surface markers. For in vivo evaluation, a cell-assisted lipotransfer animal model was employed to gauge graft volume retention and histological morphology. Analysis techniques included hematoxylin and eosin staining, Western blotting, and real-time polymerase chain reaction. RESULTS: In vitro findings suggested a slight superiority of DFATs in adipogenesis and angiogenesis compared to ASCs. In vivo tests demonstrated both cell types surpassed the control in terms of graft volume retention, with the DFATs group marginally outperforming in retention rates and the ASC group presenting a slightly enhanced graft tissue structure. CONCLUSIONS: Our study underscores the distinct advantages of DFATs and ASCs in bolstering fat graft survival, offering potentially novel insights for plastic surgeons aiming to elevate fat graft survival rates.

Delivery of microRNA-423-5p by exosome from adipose-derived stem/stromal cells inhibits DVL3 to potentiate autologous fat graft survival through adipogenesis and inflammatory response.

Autologous fat grafting represents a reconstructive technique but is limited by unstable graft retention. Based on existing reports and bioinformatics prediction, we hypothesized that delivering exosomes from human adipose-derived stem/stromal cells (hADSC-Exo) would increase fat graft survival and further explore the mechanism. hADSC-Exo were extracted and identified. An autologous fat grafting model was established using donor and recipient mice, followed by hADSC-Exo treatment. hADSC-Exo promoted the retention of autologous fat grafts in mice, along with increased adipocyte activity, angiogenesis, and decreased inflammation in grafts. Moreover, hADSC-Exo potentiated the adipose differentiation of 3T3-L1 cells, enhanced the angiogenic and migratory capacity of human umbilical vein endothelial cells, and inhibited the inflammation and viability of RAW 264.7 cells. The therapeutic effect of hADSC-Exo on fat grafting was associated with the delivery of microRNA (miR)-423-5p. Deletion of miR-423-5p in Exo impaired the function of hADSC-Exo on fat retention. miR-423-5p bound to DVL3 to suppress DVL3 expression, and DVL3 deletion promoted adipose differentiation of 3T3-L1 cells. In conclusion, our findings further widen the theoretical basis of the clinical application of hADSC-Exo in autologous fat grafts.

Increased Fat Graft Survival by Promoting Adipocyte Dedifferentiation.

BACKGROUND: Some adipocytes undergo dedifferentiation after fat transplantation, and this may affect the survival of fat grafts. However, this effect has not been adequately studied. OBJECTIVES: This study aimed to clarify the effect of promoting the dedifferentiation of mature adipocytes on the survival of fat grafts. METHODS: Mature adipocytes and adipose stem cells (ASCs) were treated with OSI-906 (a specific inhibitor of insulin receptor and insulin-like growth factor-1 receptor) in vitro, and then the dedifferentiation of mature adipocytes and the proliferation of ASCs were evaluated. In the in vivo experiment, human lipoaspirates mixed with phosphate-buffered saline (Group A) or OSI-906 (Group B) were compared in nude mice. Grafts were harvested at 2, 8, and 12 weeks, and volume retention rate, histologic, and immunohistochemical analyses were conducted. RESULTS: OSI-906 can promote the dedifferentiation of mature adipocytes and inhibit the proliferation of ASCs. At 12 weeks, Group B showed a better volume retention rate (mean [standard deviation, SD], 62.3% [7.61%]) than group A (47.75% [6.11%]) (P < .05). Moreover, viable adipocytes and vascularization showed greater improvement in Group B than in Group A. CONCLUSIONS: This study suggests that promoting the dedifferentiation of mature adipocytes can improve the survival rate and quality of fat grafts.

Self-feedback LSTM regression model for real-time particle source apportionment.

Atmospheric particulate matter pollution has attracted much wider attention globally. In recent years, the development of atmospheric particle collection techniques has put forwards new demands on the real-time source apportionments techniques. Such demands are summarized, in this paper, as how to set up new restraints in apportionment and how to develop a non-linear regression model to process complicated circumstances, such as the existence of secondary source and similar source. In this study, we firstly analyze the possible and potential restraints in single particle source apportionment, then propose a novel three-step self-feedback long short-term memory (SF-LSTM) network for approximating the source contribution. The proposed deep learning neural network includes three modules, as generation, scoring and refining, and regeneration modules. Benefited from the scoring modules, SF-LSTM implants four loss functions representing four restraints to be followed in the apportionment, meanwhile, the regeneration module calculates the source contribution in a non-linear way. The results show that the model outperforms the conventional regression methods in the overall performance of the four evaluation indicators (residual sum of squares, stability, sparsity, negativity) for the restraints. Additionally, in short time-resolution analyzing, SF-LSTM provides better results under the restraint of stability.

Washing Lipoaspirate Improves Fat Graft Survival in Nude Mice.

BACKGROUND: The optimal fat processing technique of fat grafting has not been determined. We have proved the importance of washing lipoaspirate to remove blood, but the necessity of washing when there is no obvious bleeding during liposuction is not clear. OBJECTIVES: The purpose of this study is to further investigate the effect of washing on fat graft survival and the underlying mechanisms, from the perspective of inflammation, oxidative stress and apoptosis. METHODS: To exclude the influence of blood, de-erythrocyte infranatant (dEI) isolated from lipoaspirate was obtained. Purified fat processed by cotton pad filtration mixed with dEIs after sedimentation (sedimentation group), washing (washing group) or phosphate buffer solution (control group) was transplanted to nude mice subcutaneously. Samples were harvested at 1 day and 1, 3, 8 weeks after transplantation. Volume and weight retention, histologic examination, immunostaining of perilipin-1, CD31, CD45 and Ly6g, mRNA expression of PPAR-γ, C/EBPα, VEGF, bFGF, IL-6, IL10, TNF-α, TGF-ß, Bax and Bcl-2, and protein contents of 8-iso-PGF2α, IL-6, IL10, TNF-α and TGF-ß were all compared among groups. RESULTS: After transplantation, volume and weight retention, histologic scores, viable adipocytes and vascularization were all improved in the washing group, with increased expression of adipogenic and angiogenic genes. Compared with the sedimentation group, the washing group had milder inflammation, lower levels of oxidative stress and apoptosis. CONCLUSIONS: Washing lipoaspirate to eliminate mixed components can improve fat graft survival and promote adipogenesis and angiogenesis, possibly by relieving inflammation, reducing oxidative stress injury and inhibiting apoptosis. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of 47 these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors http://www.springer.com/00266 .

Dedifferentiation of Human Adipocytes After Fat Transplantation.

BACKGROUND: Fat transplantation is a common method employed to treat soft-tissue defects. The dedifferentiation of mature adipocytes has been well documented, but whether it occurs after fat transplantation remains unclear. OBJECTIVES: The major purpose of this project was to investigate the dedifferentiation of mature adipocytes after fat transplantation. METHODS: Human lipoaspirate tissue was obtained from 6 female patients who underwent esthetic liposuction. Mature adipocytes were extracted and labeled with PKH26, mixed with lipoaspirate, and injected into nude mice. In addition, PKH26+ adipocytes were subjected to a ceiling culture. Grafted fat was harvested from nude mice, and stromal vascular fragment cells were isolated. The immunophenotype of PKH26+ cells was detected by flow cytometry analysis at 2 days and 1 week. The PKH26+ cells were sorted and counted at 2 and 4 weeks to verify their proliferation and multilineage differentiation abilities. RESULTS: Two days after transplantation, almost no PKH26+ cells were found in the stromal vascular fragment cells. The PKH26+ cells found 1 week after transplantation showed a positive expression of cluster of differentiation (CD) 90 (CD90) and CD105 and a negative expression of CD45. This indicates that the labeled adipocytes were dedifferentiated. Its pluripotency was further demonstrated by fluorescent cell sorting and differentiation culture in vitro. In addition, the number of live PKH26+ cells at week 4 [(6.83 ± 1.67) × 104] was similar with that at week 2 [(7.11 ± 1.82) × 104]. CONCLUSIONS: Human mature adipocytes can dedifferentiate into stem cell-like cells in vivo after fat transplantation.

Circular RNA expression profiles in the plasma of patients with infantile hemangioma determined using microarray analysis.

Circular RNAs (circRNAs) are noncoding RNAs that have important roles in tumor progression. Previous studies have examined the circRNAs involved in infantile hemangioma (IH) tumors. The present study compared the circRNA levels in plasma samples from patients with IH and control individuals. The circRNA expression profiles were determined using microarray in three pairs of plasma samples from patients with proliferative IH and healthy control subjects. Expression of the identified differentially expressed circRNAs was verified using reverse transcription-quantitative PCR (RT-qPCR) and a bioinformatics analysis was performed to predict the microRNAs targeted by the validated circRNAs. From the circRNA expression profiles in the plasma of patients with IHs, 128 differentially expressed circRNAs were identified, of which 72 were upregulated and 56 were downregulated. The downregulated expression of three circRNAs [Homo sapiens (hsa)_circRNA_101566, hsa_circRNA_103546 and hsa_circRNA_103573] was verified using RT-qPCR. Gene Ontology term and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that all identified networks participated in angiogenesis and tumor formation and progression. It was determined that hsa_circRNA_101566, which is able to regulate the mTOR signaling pathway, may be an important regulatory molecule in IH development and that targeting of hsa_miR_520c is able to indirectly regulate the vascular endothelial growth factor signaling pathway. Further studies are required to clarify these effects and the underlying mechanisms.

Safety of intralesional injection of lauromacrogol combined with triamcinolone for infantile hemangiomas.

The efficacy of lauromacrogol injection therapy and intralesional triamcinolone for infantile hemangiomas (IH) has been well documented recently, but with an increase in serious or rare adverse reactions. The aim of this study is to investigate the safety concerns regarding intralesional injection of lauromacrogol combined with triamcinolone for IH and to study its effect on infant growth and development. A total of 1039 IH patients who were subjected to intralesional injection of lauromacrogol combined with triamcinolone in the Plastic Surgery Department of Shandong Provincial Hospital between 1 January 2015 and 31 May 2018 were enrolled in this study. When the dose of lauromacrogol and triamcinolone was less than 3.5 and 2.0 mg/kg respectively, no serious side-effects were observed. The adverse event rate reported was 7.7%. Among the 405 patients not subjected to propranolol before the last injection, the study included three modes of treatment response: regression (82.7%), stabilization (13.8%) and failure (3.5%). By comparing height and weight to the reference standards and also by comparisons between the same-sex groups, our results confirmed that there was no significant effect on children's height and weight, regardless of whether the injection therapy was combined with oral propranolol at the appropriate dose and with more than 4-week intervals. Intralesional injection of lauromacrogol combined with triamcinolone in the treatment of IH was highly safe and effective.