Combining High-Density Fat and Condensed Low-Density Fat Injections for Precise Facial Rejuvenation.

BACKGROUND: Facial aging involves ptosis, adipose atrophy, and skeletal resorption. Depletion of adipose tissue primarily affects the deep facial fat compartment, leading to facial depression or ptosis, accompanied by atrophy of the superficial compartment. Restoring volume in the deep fat compartment is crucial for facial rejuvenation, while enhancing its supportive properties is also important. The superficial fat compartment contains small-sized adipocytes, and autologous fat grafting is a popular approach. However, variability in fat retention, homogeneity, and processing methods can impact outcomes, necessitating careful selection of a suitable fat processing material for precise facial fat grafting. METHOD: A retrospective study was conducted on 50 patients who underwent facial augmentation using combined transplantation of high-density fat (HDF) and condensed low-density fat (CLDF) and 25 patients who underwent conventional Coleman fat grafting. Coleman fat was harvested by standard technique and the adipose tissue was divided into HDF and CLDF fractions through centrifugation. Subsequently, the low-density fat fraction was subjected to a process involving physical disruption followed by additional centrifugation to obtain CLDF. The CLDF fraction was consequently injected into the pre-SMAS subcutaneous layer of the superficial fat compartments. Patient satisfaction was evaluated using a typical Likert scale. Photographs were taken and imageological examinations were performed before and after treatment. RESULT: The CLDF+HDF grafting group demonstrated a significantly shorter duration of swelling (6.0 ± 1.2 to 12.6 ± 3.3 days) and higher level of patient satisfaction when compared to the Coleman fat group. No serious complications were observed among all the patients who received the injections. CONCLUSION: The use of this new treatment approach allows for precise fat transplantation in facial regions. The use of high-concentration fat filling for deep facial layers and CLDF filling for superficial layers is a safe and effective treatment plan for facial rejuvenation. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. 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 .

The BTB/TAZ domain-containing protein CmBT1-mediated CmANR1 ubiquitination negatively regulates root development in chrysanthemum.

Roots are fundamental for plants to adapt to variable environmental conditions. The development of a robust root system is orchestrated by numerous genetic determinants and, among them, the MADS-box gene ANR1 has garnered substantial attention. Prior research has demonstrated that, in chrysanthemum, CmANR1 positively regulates root system development. Nevertheless, the upstream regulators involved in the CmANR1-mediated regulation of root development remain unidentified. In this study, we successfully identified bric-a-brac, tramtrack and broad (BTB) and transcription adapter putative zinc finger (TAZ) domain protein CmBT1 as the interacting partner of CmANR1 through a yeast-two-hybrid (Y2H) screening library. Furthermore, we validated this physical interaction through bimolecular fluorescence complementation and pull-down assays. Functional assays revealed that CmBT1 exerted a negative influence on root development in chrysanthemum. In both in vitro and in vivo assays, it was evident that CmBT1 mediated the ubiquitination of CmANR1 through the ubiquitin/26S proteasome pathway. This ubiquitination subsequently led to the degradation of the CmANR1 protein and a reduction in the transcription of CmANR1-targeted gene CmPIN2, which was crucial for root development in chrysanthemum. Genetic analysis suggested that CmBT1 modulated root development, at least in part, by regulating the level of CmANR1 protein. Collectively, these findings shed new light on the regulatory role of CmBT1 in degrading CmANR1 through ubiquitination, thereby repressing the expression of its targeted gene and inhibiting root development in chrysanthemum.

Condensing of Low-Density Fat by Mechanical Process Improves Fat Retention and Reduces Oil Cyst Formation in Breast Reconstruction.

BACKGROUND: Although autologous fat grafting is a useful adjunct for breast reconstruction, its indications remain limited as large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated the volumization effect and complications with combined fat grafting of condense low-density fat and high-density fat. METHODS: This retrospective, single-center study included 25 patients who underwent combined grafting of condensed low-density fat and high-density fat (CLDF + HDF) and 20 patients who underwent conventional Coleman fat grafting for breast reconstruction from December 2017 to January 2022. Retention rates and complications were evaluated by magnetic resonance imaging and ultrasound rates. Patient satisfaction was evaluated using a typical Likert scale. Photographs were taken and imageological examinations were performed before and after treatment. OUTCOMES: Graft retention rate was higher in patients who underwent CLDF + HDF than Coleman fat grafting for breast reconstruction (38.40 ± 4.41% vs. 31.43 ± 5.43%, p <0.05). One patient in the CLDF + HDF grafting group, compared with twelve in the Coleman fat grafting group, developed oil cysts exceeding 1 cm. Patient satisfaction rate was higher in the CLDF + HDF grafting group. CONCLUSIONS: Mechanical processes can concentrate the cellular content of LDF and remove oil, condensing LDF to the level of HDF. Combined grafting of CLDF optimized by mechanical processing and HDF is effective for breast reconstruction, with a higher retention rate and a lower incidence of complications than conventional Coleman fat grafting. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. 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 .

DIAPH3 is a prognostic biomarker and inhibit colorectal cancer progression through maintaining EGFR degradation.

BACKGROUND: Actin cytoskeleton is connected with the processes of cell proliferation and migration in colorectal cancer (CRC). However, it is unknown how to accomplish these adjustments in CRC by actin cytoskeleton genes (ACGs) and here we investigated the role of hub prognosis-related ACGs-Diaphanous-related formin 3 (DIAPH3) in CRC, as a potential, novel target. METHODS: The ACGs gene set from the Kyoto Encyclopedia of Genes and Genomes (KEGG) was used to group CRC patients and select prognosis-related ACGs by univariate and multivariate Cox regression for constructing prognostic model. Next, we tested hub prognosis-related ACGs- DIAPH3 expression in CRC and clarified the role of DIAPH3 by shRNA constructs in KM12 and SW480. Activation of EGFR was analyzed by western blot and immunofluorescence. RESULTS: The results showed that actin cytoskeleton function is a significant prognostic factor for CRC patients and related to clinicopathological characteristics such as T stage and lymph node metastasis. A prognostic model constructed by four prognosis-related ACGs has a moderate intensity to 1-year Survival (AUC = 0.71). And hub prognosis-related ACGs DIAPH3 is downregulated in CRC. Knockdown of DIAPH3 could promote the proliferation and migration capacity of CRC. In addition, DIAPH3-silenced cells increase EGFR phosphorylation by inhibiting EGFR transportation to lysosome. CONCLUSIONS: ACGs play a significant role in tumor invasion and have the potential to predict the prognosis of CRC. Prognosis-related ACGs DIAPH3 might be a new prognostic biomarker and DIAPH3 could inhibit CRC progression through maintaining EGFR degradation.

Improving Low-Density Fat by Condensing Cellular and Collagen Content through a Mechanical Process: Basic Research and Clinical Applications.

BACKGROUND: Large-volume fat grafting results in high absorption and complication rates. Low-density fat includes small numbers of viable cells and considerable oil, resulting in nodules and oil cysts. This study evaluated a strategy for large-volume fat grafting using a mechanical process to condense low-density fat and transplanting it with high-density fat. METHODS: Low-density fat, defined as the upper half of centrifuged lipoaspirates, was emulsified by intersyringe shifting and centrifuged to obtain condensed low-density fat. Fresh condensed low-density fat was analyzed by counting cells in the stromal vascular fraction, and by electron scanning and Western blotting. The retention rate and histologic changes of the product were analyzed using a fat grafting model in nude mice. Transplantation with a combination of condensed low-density fat and high-density fat was tested in patients undergoing breast reconstruction and breast augmentation. RESULTS: The condensed low-density fat derived from low-density fat contained a large number of stromal vascular fraction cells and collagens, comparable to that of high-density fat and much higher than in low-density fat and Coleman fat. Retention rates 12 weeks after transplantation were higher for condensed low-density fat (55.0 ± 7.5 percent) than for low-density fat (31.1 ± 5.7 percent) and Coleman fat (41.1 ± 6.8 percent), with condensed low-density fat having fewer oil cysts and lower macrophage infiltration. Patients grafted with combined condensed low-density fat and high-density fat showed good long-term volume retention. CONCLUSIONS: Using mechanical methods to condense low-density fat to a level comparable to that of high-density fat is a practical method of improving fat graft retention and avoiding severe complications. This new strategy may improve the quality of lipoaspirates for patients requiring large-volume augmentation.

Induced Beige Adipocytes Improved Fat Graft Retention by Promoting Adipogenesis and Angiogenesis.

BACKGROUND: Fat grafting is commonly used in treating soft-tissue defects. However, the basic biology behind fat grafting is still not fully understood. Evidence of adipose browning into beige adipose tissue after fat grafting was revealed, but its role in fat grafting remains unclear. METHODS: Induced beige adipocytes and adipose-derived stem cells were obtained from human lipoaspirates and labeled with green fluorescent protein. Nude mice were each injected with 300 mg of human lipoaspirate containing green fluorescent protein-labeled adipose-derived stem cells, green fluorescent protein-labeled induced beige adipocytes, or phosphate-buffered saline. Grafted fat was harvested after 1, 4, 8, and 12 weeks for immunohistochemistry and histologic examination. Graft retention, vascularization, and adipogenic gene expression were compared. RESULTS: After 7 days' induction, adipocytes achieved browning with multilocular lipid droplets, increased mitochondria, and up-regulated browning gene expression. Fat graft retention rates at week 12 were significantly higher after injection of induced beige adipocytes than after injection of phosphate-buffered saline (46.0 ± 4.9 percent versus 31.0 ± 3.6 percent; p = 0.01), but were similar after injection of induced beige adipocytes and adipose-derived stem cells (p > 0.05). Induced beige adipocytes underwent rewhitening into white adipocytes and showed up-regulation of peroxisome proliferator-activated receptor-γ expression. Induced beige adipocytes enhanced angiogenesis, but were not active in forming vessel structures. CONCLUSIONS: Induced beige adipocytes and adipose-derived stem cells were comparable in improving fat graft retention rates. Induced beige adipocytes promote angiogenesis in a paracrine manner and are prone to rewhitening after fat grafting.

Mechanical Micronization of Lipoaspirates for Regenerative Therapy.

Stromal vascular fraction (SVF) has become a regenerative tool for various diseases; however, legislation strictly regulates the clinical application of cell products using collagenase. Here, we present a protocol to generate an injectable mixture of SVF cells and native extracellular matrix from adipose tissue by a purely mechanical process. Lipoaspirates are put into a centrifuge and spun at 1,200 x g for 3 min. The middle layer is collected and separated into two layers (high-density fat at the bottom and low-density fat on the top). The upper layer is directly emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance under the oil layer is collected and defined as the extracellular matrix (ECM)/SVF-gel. The oil on the top layer is collected. Approximately 5 mL of oil is added to 15 mL of high-density fat and emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance is also ECM/SVF-gel. After the transplantation of the ECM/SVF-gel into nude mice, the graft is harvested and assessed by histologic examination. The result shows that this product has the potential to regenerate into normal adipose tissue. This procedure is a simple, effective mechanical dissociation procedure to condense the SVF cells embedded in their natural supportive ECM for regenerative purposes.

Ki-67 is an independent prognostic marker for the recurrence and relapse of oral squamous cell carcinoma.

As a nuclear and nucleolar protein, proliferation marker protein Ki-67 (Ki-67) serves a vital role in tumorigenesis due to its positive correlation with tumor proliferation. High expression of Ki-67 in the cell cycle from the G1 to M phase makes it a potential biomarker for certain tumors and useful for selecting medical treatment. However, the diagnostic value of Ki-67 in oral squamous cell carcinoma (OSCC) has not been fully evaluated. In the present study, the objective was the elucidation of the prognostic value of Ki-67 in a large number of OSCC patients. Ki-67 expression was detected by immunohistochemical staining methods in 298 OSCC specimens and 98 tumor-free oral mucosa specimens (62 dysplasia mucosa and 36 normal mucosa), acquired from Nanjing Stomatological Hospital, Medical School of Nanjing University (Nanjing, China). Expression of Ki-67 in normal tissues, dysplasia tissues and OSCC tissues was compared. Associations between Ki-67 expression and clinicopathological parameters were analyzed by χ2 test. Kaplan-Meier survival curves and Cox progression analysis were used to assess the diagnostic value of Ki-67 for OSCC. The results showed that Ki-67 expression was higher in OSCC tissues than in tumor-free tissues and that it increased with the progression of dysplasia in oral mucosa tissues. In addition, patients with high Ki-67 expression had a worse clinical outcome, including poor tumor differentiation (P=0.001), increased positive lymph node metastasis (P=0.006) and increased worst pattern of invasion type (P<0.0001). Kaplan-Meier survival analysis demonstrated that higher Ki-67 expression was associated with poorer overall survival (OS) (P=0.035), recurrence-free survival (RFS) (P=0.017), metastasis-free survival (MFS) (P=0.032) and disease-free survival (DFS) (P=0.018) times. Additional multivariate analysis demonstrated that Ki-67 expression was negatively associated with OS, DFS, RFS and MFS. In conclusion, Ki-67 overexpression is associated with the progression of OSCC and serves as an independent prognostic factor for OSCC patients.

Frequency of circulating CD14++ CD16+ intermediate monocytes as potential biomarker for the diagnosis of oral squamous cell carcinoma.

BACKGROUND: Monocytes, which subdivided into three functional subsets (classical, intermediate, and nonclassical), play important roles in the progression of cancer. The subset composition is altered in several pathologic conditions including cancers. However, the composition and function of circulating monocyte subsets in patients with oral squamous cell carcinoma (OSCC) are still obscure. METHODS: The frequencies of monocyte subsets in peripheral blood of patients with OSCC and healthy donors are determined by flow cytometry, and their diagnostic values for OSCC were evaluated. The associations between levels of monocyte subsets and clinicopathological features of patients with OSCC were analyzed using cross-tabulation with the chi-square test. RESULTS: We demonstrated that the frequency of CD14++ CD16+ intermediate monocytes was remarkably increased (P < 0.0001) in OSCC patients compared with healthy controls (7.33% ± 2.56% of total monocytes, n = 68 versus 4.78% ± 1.50% of total monocytes, n = 57). A trend of decrease in CD14++ CD16- classical subset was observed between these two groups (P = 0.0508), whereas no significant difference was detected in CD14+ CD16++ nonclassical subset (P > 0.05). The receiver operating characteristic (ROC) curve analysis indicated that the frequency of intermediate monocytes (AUC = 0.810, P < 0.0001) could be a potential diagnostic biomarker to discriminate patients with OSCC from healthy subjects. Moreover, this parameter was significantly correlated to the worst pattern of invasion (WPOI, P < 0.05) of OSCC tissues. CONCLUSIONS: Detection of monocyte subsets in peripheral blood sheds a light on utilizing the frequency of intermediate monocytes as a potential diagnostic biomarker for OSCC.

Calix[8]arene Sulfonic Acid Catalyzed Three-Component Reaction for Convenient Synthesis of 3,4-Dihydropyrimidin-2(1H)-ones/thiones under Ultrasonic Irradiation.

In this work, the catalytic activity of calix[8]arene sulfonic acid was successfully investigated for the famous Biginelli reaction. Under ultrasonic irradiation, calix[8]arene sulfonic acid could efficiently catalyzed the three-component reaction of aldehydes with ethyl acetoacetate and urea or thiourea in ethanol to afford the corresponding 3,4-dihydropyrimidin-2(1H)-ones/thiones in 46-93%. The advantages of this method are the easy isolated procedure, short reaction time and low cost of the catalyst.

Preparation and characterization of amphiphilic calixarene nanoparticles as delivery carriers for paclitaxel.

Two types of amphoteric calix[n]arene carboxylic acid (CnCA) derivative, i.e., calix[6]arene hexa-carboxylic acid (C6HCA) and calix[8]arene octo-carboxylic acid (C8OCA), were synthesized by introducing acetoxyls into the hydroxyls of calix[n]arene (n=6, 8). C6HCA and C8OCA nanoparticles (NPs) were prepared successfully using the dialysis method. CnCA NPs had regular spherical shapes with an average diameter of 180-220 nm and possessed negative charges of greater than -30 mV. C6HCA and C8OCA NPs were stable in 4.5% bovine serum albumin solutions and buffers (pH 5-9), with a low critical aggregation concentration value of 5.7 mg·L(-1) and 4.0 mg·L(-1), respectively. C6HCA and C8OCA NPs exhibited good paclitaxel (PTX) loading capacity, with drug loading contents of 7.5% and 8.3%, respectively. The overall in vitro release behavior of PTX from the CnCA NPs was sustained, and C8OCA NPs had a slower release rate compared with C6HCA NPs. These favorable properties of CnCA NPs make them promising nanocarriers for tumor-targeted drug delivery.

Therapeutic efficacy of a modified method with splints in correction of cryptotia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the therapeutic efficacy of a modified method with splints in correction of cryptotia.</p><p><b>METHODS</b>From Oct. 2012 to Jan. 2014, 3 cases with unilateral cryptotia were treated with the modified method with splints. The muscles attached to the periosteum of cartilage were dissected. The muscles between the ear and temper were cut off. Then one silicon tube was put around the ear through cranioauricular sulcus. The other tube was placed within the cavum conchae and fixed with the tube around the ear.</p><p><b>RESULTS</b>No flap necrosis happened. The patients were followed up for six months to one year with satisfied and stable results.</p><p><b>CONCLUSIONS</b>The modified method is easy to perform with less morbidity. It is one of the ideal correction for cryptotia.</p>

[Lux1540nm fractional laser treatment for superficial scar].

OBJECTIVE: To observe the effect of non-ablative fractional laser Lux1540nm in superficial scars therapy. METHODS: According to the Fitzpatrik skin type and scar appearances, appropriate laser energy and pulse width were selected for scar treatment. Therapeutic effect and side-effect was evaluated by preoperative photos and 6 months of postoperative photos. RESULTS: The total effective rate was 78.12% (28/32). 7 cases (21.88%) showed ineffective results. The therapeutic effect was improved along the increase of treatments. Temporary hyperpigmentation was found in 3 cases (9.38%) and faded away within 3-6 months. Temporary erythema faded away within 4-24 hours. CONCLUSION: The treatment of non-ablative fractional laser Lux1540nm for superficial scar is effective with minimum side effect.