Study on the estradiol degradation gene expression and resistance mechanism of Rhodococcus R-001 under low-temperature stress.

Estradiol (E2), an endocrine disruptor, acts by mimicking or interfering with the normal physiological functions of natural hormones within organisms, leading to issues such as endocrine system disruption. Notably, seasonal fluctuations in environmental temperature may influence the degradation speed of estradiol (E2) in the natural environment, intensifying its potential health and ecological risks. Therefore, this study aims to explore how bacteria can degrade E2 under low-temperature conditions, unveiling their resistance mechanisms, with the goal of developing new strategies to mitigate the threat of E2 to health and ecological safety. In this paper, we found that Rhodococcus equi DSSKP-R-001 (R-001) can efficiently degrade E2 at 30 °C and 10 °C. Six genes in R-001 were shown to be involved in E2 degradation by heterologous expression at 30 °C. Among them, 17ß-HSD, KstD2, and KstD3, were also involved in E2 degradation at 10 °C; KstD was not previously known to degrade E2. RNA-seq was used to characterize differentially expressed genes (DEGs) to explore the stress response of R-001 to low-temperature environments to elucidate the strain's adaptation mechanism. At the low temperature, R-001 cells changed from a round spherical shape to a long rod or irregular shape with elevated unsaturated fatty acids and were consistent with the corresponding genetic changes. Many differentially expressed genes linked to the cold stress response were observed. R-001 was found to upregulate genes encoding cold shock proteins, fatty acid metabolism proteins, the ABC transport system, DNA damage repair, energy metabolism and transcriptional regulators. In this study, we demonstrated six E2 degradation genes in R-001 and found for the first time that E2 degradation genes have different expression characteristics at 30 °C and 10 °C. Linking R-001 to cold acclimation provides new insights and a mechanistic basis for the simultaneous degradation of E2 under cold stress in Rhodococcus adaptation.

Aesthetic Improvement of Body Proportion Through Trunk Liposculpture: A Waistline-based Strategy.

BACKGROUND: Abdominal contouring through liposuction has been practiced for decades. However, few studies have focused on describing the definition and enhancement of the waistline in torso contouring procedures. OBJECTIVES: In the present study, the authors proposed a waistline-based strategy for abdominal liposculpture to achieve a better aesthetic outcome and emphasize high overall patient satisfaction. METHODS: The data of patients who underwent the waistline-based liposculpture procedure from 2020 to 2023 were retrospectively reviewed. Aesthetic improvement of the central trunk contour was evaluated and analyzed by comparing preoperative and postoperative photogrammetric measurements. Satisfaction with the outcome was assessed with a patient satisfaction questionnaire. RESULTS: A total of 70 patients were enrolled in this study. During 6 months of postoperative evaluation, the shape of the central trunk contour improved significantly (both waist concavity and hip convexity increased quantitatively, P < .05), while the position of the waist did not differ significantly postoperatively (P > .05). All patients were satisfied with their postoperative outcomes, including their overall aesthetic appearance, waistline position, and waist-to-hip ratio. There were no intraoperative complications or rare postoperative complications. CONCLUSIONS: Waistline-based liposculpture is a simple and effective procedure to improve the aesthetic outcomes of trunk contouring and has highly satisfactory results after long-term follow-up.

A rare case of hepatoid adenocarcinoma of the lung.

Hepatoid adenocarcinoma of the lung is a special type of primary origin in the lung with obvious pathological features and short survival time. However, standard treatment guidelines have not yet been established. Herein, we report a case of hepatoid adenocarcinoma with the primary lesion located in the left upper lung. The tumour size was reduced after four cycles of combined therapy. Subsequent postoperative pathology confirmed complete remission.

Application of a Proposed Multi-Positional Circumferential Arm Liposuction Method and Quantification of its Clinical Efficacy Evaluation.

BACKGROUND: Upper arm liposuction mainly focuses on the posterolateral region, which may lead to a lack of harmony between the aspirated and unaspirated areas. In addition, the treatment effect of arm liposuction is often evaluated only by preoperative and postoperative photograph comparison and simple measurement; quantitative research on this topic is still lacking. METHODS: The multi-positional circumferential arm liposuction (MCAL) technique was proposed and applied to a total of 34 females in our hospital from 2017 to 2019. Three-dimensional data of 12 patients before the operation and after 2-3 months were collected and processed by 3D imaging, and the volume reduction rate was evaluated quantitatively. RESULTS: The MCAL method was successfully applied in the clinic, and its surgical effect was quantitatively studied. The mean follow-up time of 12 patients was (75.2 ±13.1) days, and the postoperative volume was significantly reduced. The postoperative volume of patients with type I, type II and type III decreased by (10.79 ±2.55)%, (17.25 ±3.02)% and (22.76 ±3.51)%, respectively. CONCLUSION: Our new MCAL technique was successful, maximizing the esthetic results in upper limb contour refinements in the superficial fascial layer. The clinical efficacy of this proposed MCAL method was evaluated by CT and 3D digital technology, which provided further accuracy in demonstrating its effect on the shape of the arm. 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 https://www.springer.com .

Inhibition of microRNA-21-5p reduces keloid fibroblast autophagy and migration by targeting PTEN after electron beam irradiation.

Electron beam (EB) irradiation is useful to reduce the recurrence of keloids; however, the underlying mechanism remains unknown. MicroRNA-21 (miR-21), which regulates autophagy during cancer radiation therapy, was identified as a potential therapeutic target for keloids. Here, we investigate the regulatory mechanism(s) of miR-21-5p on keloid fibroblast autophagy and migration after EB irradiation. The microRNA expression profile of the keloid dermis was examined by performing a microRNA microarray. Levels of LC3B and Beclin-1 were detected by immunohistochemical and western blot analysis in the keloid dermis and fibroblasts. Autophagy and apoptosis were tested in keloid fibroblasts after EB irradiation or transfection with an miR-21-5p inhibitor using electron microscopy, a Cyto-ID Green Autophagy Detection Kit, and an Annexin V PE Apoptosis Detection Kit. Migration was analyzed by an in vitro scratch-wound healing assay. Mechanistic tests were performed using small interfering RNAs to phosphatase and tensin homolog (siPTEN). Levels of miR-21-5p, PTEN, programmed cell death 4 (PDCD4), p-AKT, and apoptosis- and autophagy-associated genes were examined by qRT-PCR and western blotting. LC3B expression and migration ability were enhanced in fibroblasts and the keloid margin dermis compared with those in the adjacent normal skin. Both EB irradiation and an miR-21-5p inhibitor reduced keloid fibroblast autophagy, which was accompanied by decreased expression of miR-21-5p, p-AKT, and LC3B-II and increased expression of PTEN, PDCD4, and apoptosis-related genes. MiR-21-5p downregulation inhibited migration and suppressed LC3B expression and this was reversed by PTEN reduction. In conclusion, with increasing apoptosis, EB irradiation inhibits autophagy in keloid fibroblasts by reducing miR-21-5p, which regulates migration and LC3B expression via PTEN/AKT signaling. These data suggest a potential mechanism wherein miR-21-5p inhibition regulates autophagy and migration in EB-irradiated keloid fibroblasts, effectively preventing local invasion and recurrence. Therefore, miR-21-5p could be a new therapeutic target, to replace EB irradiation, and control keloid relapse.

Systematic Analysis of Clinical Outcomes of Bilateral Sagittal Split Ramus Osteotomy With Preoperative Modeling and Postoperative Orthodontic in the Treatment of Mandibular Protrusion.

The purpose of this study was to determine the changes in hard tissues, soft tissues, and teeth after bilateral sagittal split ramus osteotomy and orthodontic treatment for the treatment of mandibular protrusion. Cephalometric analysis was used to evaluate the aesthetic effects and occlusal relationships obtained. The subjects included 11 women and 9 men (aged 18-27 years; average, 20 years) with mandibular protrusion who underwent bilateral sagittal split ramus osteotomy. Based on a preoperative computer-aided manufacturing/design-assisted, model surgical design and an occlusal guide plate, new occlusal relationships were established for the patients. In addition, the preoperative and the end of postoperative orthodontic treatment cephalometric radiographs were systematically analyzed. In all patients, the surgical incisions underwent primary healing, with no infection or osteonecrosis. Significant differences were observed in the preoperative and the end of postoperative orthodontic treatment values of all hard tissue and teeth parameters, except for SNA°, ANB°, GoGn-SN°, SE (mm), NP-FH°, SGn-FH°, OP-FH°, 1-MP°, Li-E (mm). The most obvious significant differences were seen in SNB°, SND°, 1_-NA°, 1_-NA (mm), 1-NB (mm), 1-NB°, Po-NB (mm), NA-PA°, AB-NP°, 1-OP°, Ui-E (mm), and S-N'-B'° (P < 0.001). Postoperative follow-up lasted for 10 to 12 months. All patients eventually achieved normal downjaw relationship, tooth arch forms, and spee curves. There were no evident irregularities of teeth arrangement or abnormal occlusal relationships were observed. All patients were satisfied with their facial appearance and occlusal relationships at the end of postoperative orthodontic treatment. The authors found a precise preoperative model surgical design combined with postoperative orthodontic treatment is a simple and time-saving technique. It can be used to correct mandibular protrusion with satisfactory occlusal relationship, facial appearance, and minimal postoperative complications.

MiR-21-5p Links Epithelial-Mesenchymal Transition Phenotype with Stem-Like Cell Signatures via AKT Signaling in Keloid Keratinocytes.

Keloid is the abnormal wound healing puzzled by the aggressive growth and high recurrence rate due to its unrevealed key pathogenic mechanism. MicroRNAs contribute to a series of biological processes including epithelial-mesenchymal transition (EMT) and cells stemness involved in fibrotic disease. Here, using microRNAs microarray analysis we found mir-21-5p was significantly up-regulated in keloid epidermis. To investigate the role of miR-21-5p in keloid pathogenesis, we transfected miR-21-5p mimic or inhibitor in keloid keratinocytes and examined the abilities of cell proliferation, apoptosis, migration and invasion, the expressions of EMT-related markers vimentin and E-cadherin and stem-like cells-associated markers CD44 and ALDH1, and the involvement of PTEN and the signaling of AKT and ERK. Our results demonstrated that up-regulation or knockdown of miR-21-5p significantly increased or decreased the migration, invasion and sphere-forming abilities of keloid keratinocytes, and the phenotype of EMT and cells stemness were enhanced or reduced as well. Furthermore, PTEN and p-AKT were shown to participate in the regulation of miR-21-5p on EMT phenotypes and stemness signatures of keloid keratinocytes, which might account for the invasion and recurrence of keloids. This molecular mechanism of miR-21-5p on keloid keratinocytes linked EMT with cells stemness and implicated novel therapeutic targets for keloids.

[In vitro study of effects and mechanism of rapamycin-induced autophagy in keloid fibroblasts].

Objective: To investigate the effect of rapamycin on biological characteristics and autophagy of keloid fibroblasts, and the regulation of rapamycin in mTOR (mammalian target of rapamycin) signaling pathway and autophagy-related non-coding RNAs in keloid fibroblasts. Methods: After Keloid fibroblasts were treated with rapamycin (10、50、100 nmol/L), and MTS assay was used to test the cell proliferation. The apoptosis of cells was tested by the flow cytometry analysis. The formation of autophagy was observed by TEM, and the Western Blot was used to detect the expression of autophagy-related protein LC3.Real-time PCR was used to detect the expression of genes of involued in mTOR pathway and autophagy-related non-coding RNAs. Statistical significance was determined using Paired-Samples t Test,P value less than 0.05 was considered statistically significant. Results: The ratio of 490nm was decreased significantly in rapamycin-treated keloid fibroblasts compared with that in untreated cells (P ＜ 0.05).Meanwhile the mRNA expressions of extracellular matrix (ECM) genes, including collagen-1 、α-SMA and Fibronectin, were inhibited by rapamycin (P ＜ 0.05).The flow cytometry analysis showed that the percent of apoptosis cells was not increased in rapamycin-induced cells (P ＞ O.05). The double-layer membrane structure of autophagosomes could be observed under the TEM in rapamycin-treated fibroblasts, accompanied by the increased expression of autophagy-related protein LC3.The mRNA expressions of downstream genes of mTOR pathway,4EBP1 and p70S6K,were down-regulated in rapamycin-treated fibroblasts, while the expressions of autophagy-related miRNAs, including miR-885-3p,miR-204,miR-101,miR-376b and lncRNA FLJ11812 were enhanced, and miR-30a,lncRNA HULC5 was decreased in rapamycin-treated fibroblasts (P ＜ 0.05). Conclusions: Rapamycin could inhibit the proliferation of keloid fibroblasts, and could not affect the apoptosis of cells.However, rapamycin induced the autophagy of keloid fibroblasts through regulating the expression of autophagy-related non-coding RNAs and genes in the mTOR signaling pathway.

[In vitro study of TGF-ß1-induced epithelial-mesenchymal transition of keloid epithelial cells].

OBJECTIVE: To construct and characterize the TGF-ß1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction. METHODS: The epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-ß1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant. RESULTS: The mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-ß1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-ß1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05). CONCLUSIONS: TGF-ß1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-ß1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.

Epithelial-mesenchymal transition in keloid tissues and TGF-ß1-induced hair follicle outer root sheath keratinocytes.

Keloid is a skin fibrotic disease with the characteristics of recurrence and invasion, its pathogenesis still remains unrevealed. The epithelial-mesenchymal transition (EMT) is critical for wound healing, fibrosis, recurrence, and invasion of cancer. We sought to investigate the EMT in keloid and the mechanism through which the EMT regulates keloid formation. In keloid tissues, the expressions of EMT-associated markers and transforming growth factor (TGF)-ß1/Smad3 signaling were examined by immunohistochemistry. In the keloid epidermis and dermal tissue, the expressions of genes related to the regulation of skin homeostasis, fibroblast growth factor receptor 2 (FGFR2) and p63, were analyzed using quantitative real-time polymerase chain reaction. The results showed that accompanying the loss of the epithelial marker E-cadherin and the gain of the mesenchymal markers fibroblast-specific protein 1 (FSP1) and vimentin in epithelial cells from epidermis and skin appendages, and in endothelial cells from dermal microvessels, enhanced TGF-ß1 expression and Smad3 phosphorylation were noted in keloid tissues. Moreover, alternative splicing of the FGFR2 gene switched the predominantly expressed isoform from FGFR2-IIIb to -IIIc, concomitant with the decreased expression of ΔNp63 and TAp63, which changes might partially account for abnormal epidermis and appendages in keloids. In addition, we found that TGF-ß1-induced hair follicle outer root sheath keratinocytes (ORSKs) and normal skin epithelial cells underwent EMT in vitro with ORSKs exhibiting more obvious EMT changes and more similar expression profiles for EMT-associated and skin homeostasis-related genes as in keloid tissues, suggesting that ORSKs might play crucial roles in the EMT in keloids. Our study provided insights into the molecular mechanisms mediating the EMT pathogenesis of keloids.

Ectopic osteogenesis of allogeneic bone mesenchymal stem cells loading on ß-tricalcium phosphate in canines.

BACKGROUND: Bone mesenchymal stem cells are progenitor cells for mesenchymal tissues. The objective of this study was to evaluate the subcutaneous ectopic osteogenesis of allogeneic bone mesenchymal stem cells, which were loaded on ß-tricalcium phosphate in canines without immunosuppressive therapy. METHODS: Osteoinduced allogeneic bone mesenchymal stem cells were seeded onto a ß-tricalcium phosphate scaffold to construct tissue-engineered bone. Four dogs (recipients) in the allogeneic group were subcutaneously implanted with the allogeneic bone mesenchymal stem cells/scaffold; four dogs (donors) in the autogeneic group were implanted with the autogeneic bone mesenchymal stem cells/scaffold complex; and four dogs in the control group were implanted with scaffold alone. Systemic immune responses were evaluated by measuring the T-lymphocyte CD4, CD8, and CD4/CD8 subsets of each group. Subcutaneous osteogenesis was compared between the three groups by histologic analysis at week 24 after implantation. RESULTS: Flow cytometry showed no significant differences in the number of CD4 and CD8 T cells and the CD4/CD8 T-cell ratios among the three groups. Histologically, at week 24, both the autogeneic and allogeneic complexes led to subcutaneous osteogenesis, whereas the control group alone did not. There were no significant differences in the percentage of osteogenic area between the allogeneic and the autogeneic complexes on histomorphometric analysis (p > 0.05), which was significantly higher than that produced by the control group alone (p < 0.001). CONCLUSION: The results demonstrate that osteoinduced, allogeneic bone mesenchymal stem cells loaded on ß-tricalcium phosphate enhanced ectopic bone formation in canines without immunosuppressive therapy.

Systematic analysis of clinical outcomes of anterior maxillary and mandibular subapical osteotomy with preoperative modeling in the treatment of bimaxillary protrusion.

The purpose of this study was to determine the changes in teeth and hard tissues after preoperative modeling and bimaxillary anterior subapical osteotomy for the treatment of bimaxillary protrusion. Cephalometric analysis was used to evaluate the aesthetic effects and occlusal relationships obtained. The subjects included 19 women and 1 man (aged 19-41 years; average, 29 years) with bimaxillary protrusion who underwent anterior subapical osteotomy of both the maxilla and mandible, with simultaneous genioplasty, if required. Based on a preoperative computer-aided manufacturing/design-assisted and model surgical design and an occlusal guide plate, new occlusal relationships were established for the patients. In addition, the preoperative and postoperative cephalometric radiographs were systematically analyzed. In all patients, the surgical incisions underwent primary healing, with no infection or osteonecrosis. Significant differences were observed in the preoperative and postoperative values of all hard tissue and teeth parameters, except for SGn-FH degrees and Co-MP. The most obvious significant differences were seen in L1-OP°, Id-Pog-Go°, IIA°, U1E-Apog, L1E-Apog, U1E-NA, and L1-NA° (P < 0.001). Postoperative follow-up lasted for 12 to 36 months. All patients eventually achieved normal jaw relationships, tooth arch forms, and Spee curves. No evident irregularities of teeth arrangement or abnormal occlusal relationships were observed. All patients were satisfied with their postoperative facial appearance, except for 1 patient, who underwent repeat surgery because of relapse. With the use of a precise preoperative model surgical design, orthognathic surgery, a simple and time-saving technique, can be used to correct bimaxillary protrusion with satisfactory postoperative occlusal relationship and facial aesthetic appearance and minimal postoperative complications.