Poly-p-dioxanone Thread Leads to Fat Metabolism Around the Thread in Pig Subcutaneous Back Fat.

Background: When poly-p-dioxanone (PDO) thread is implanted subcutaneously, in addition to collagen hyperplasia, it can also cause denaturation of surrounding adipocytes and reduce the thickness of the fat layer. Hitherto, no studies have thoroughly investigated the effects of PDO thread on adipose tissue. Objectives: In this study, the effect of PDO thread on adipose tissue was investigated in an animal model. Methods: In the current study, PDO thread was implanted into subcutaneous adipose tissue of the back in a miniature pig. Implantation site tissue and control site tissue were taken 12 weeks after implantation for hematoxylin and eosin (H&E) staining and transcriptome sequencing. Gene ontology functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis were performed to investigate the differential gene expression between PDO thread implantation and control site tissue. Results: An obvious decrease in the number, fusion, and denaturation of adipocytes can be seen by H&E staining. Sequencing analysis results showed that many of the genes identified, which were downregulated after PDO thread implantation, were involved in functions and pathways related to lipid metabolism, such as fatty acid metabolism, fatty acid degradation, and lipid cell lipolysis regulation. Some genes related to fatty acid metabolism were significantly downregulated in the PDO tissue at 12 weeks compared to the control tissue. Conclusions: Our results showed PDO thread implantation can cause a decrease in the number of adipocytes, as well as a significant alteration of the expression levels of some genes involved in lipid metabolism-related pathways. PDO thread might play an important role in promoting lipolysis.

Controlled-diffusion centrifugal microfluidic for rapid antibiotic susceptibility testing.

The abuse of antibiotics has become a global public safety issue, leading to the development of antimicrobial resistance (AMR). The development of antimicrobial susceptibility testing (AST) is crucial in reducing the growth of AMR. However, traditional AST methods are time-consuming (e.g., 24-72 h), labor-intensive, and costly. Here, we propose a controlled-diffusion centrifugal microfluidic platform (CCM) for rapid AST to obtain highly precise minimum inhibitory concentration (MIC) values. Antibiotic concentration gradients are generated by controlled moving and diffusing of antibiotic and buffer solution along the main microchannel within 3 min. The solution and bacterial suspension are then injected into the outermost reaction chamber by simple centrifugation. The CCM successfully determined the MIC for three commonly used antibiotics in clinical settings within 4-9 h. To further enhance practicality, reduce costs, and meet point-of-care testing demands, we have developed an integrated mobile detection platform for automated MIC value acquisition. The proposed CCM is a simple, low-cost, and portable method for rapid AST with broad clinical and in vitro applications.

Experimental investigation of biostimulatory effects after polydioxanone thread insertion in a pig model.

BACKGROUND: Polydioxanone (PDO) threads have been widely used to tighten and lift the facial soft tissue. OBJECTIVE: This research aims to determine the collagenation and inflammation changes that occur in the adipose tissue over time when different types of threads are implanted. METHODS & MATERIALS: Three threads types, PDO, poly glycolic-co-lactic acid (PGLA), and nylon, were inserted in the subcutaneous fat of 12-month-old Bama miniature pigs. Collagen production and inflammatory response were evaluated by hematoxylin and eosin and Masson trichrome staining at 1, 4, 12, 24, and 48 weeks. RESULTS: The integrity of the PDO thread lasted up to 24 weeks with mild inflammation and collagen production. The PGLA thread integrity lasted until 12 weeks and had a strong inflammatory response. The nylon thread's integrity was maintained for 48 weeks and showed minor inflammation and collagen production. CONCLUSION: Our data suggest that PDO thread is the best choice for clinicians, as it has a mild action process with minimal irritation, moderate collagen production, a reasonable explanation time, with obvious bridging fibrous tissue, and thickening action for the superficial fascia.

[Study on the influence of buried thread nasal augmentation on dorsal soft tissue of nose and revision rhinoplasty].

Objective: To investigate the influence of buried thread nasal augmentation on dorsal soft tissue of nose and revision rhinoplasty. Methods: A clinical data of 29 patients requesting revision rhinoplasty after buried thread nasal augmentation, who were admitted between July 2017 and July 2019 and met the selection criteria, was retrospectively analyzed. All patients were female with an average age of 26.8 years (range, 18-43 years). The patiens were admitted to the hospital at 3-48 months after buried thread nasal augmentation (median, 15 months). Among them, there were 18 cases of insufficient nasal tip projection, 22 cases of insufficient nasal root projection, 7 cases of threads ectasia, 5 cases of threads exposure, 3 cases of infection, and 10 cases with two or more conditions. There were 9 cases of combined short nose deformity, 1 case of spherical hypertrophy of the nasal tip, 3 cases of deviation of the nasal columella, 3 cases of excessive width of the nasal base, and 1 case of nasal hump. Three infected patients only underwent threads removal and debridement. The rest patients underwent revision rhinoplasty, and the dorsum of the nose was made with polytetrafluoroethylene expansion; the tip of the nose was reshaped by taking autologous rib cartilage and alar cartilage in 16 cases, and by taking autologous septal cartilage and alar cartilage in another 10 cases. The threads and surrounding tissue specimens removed during operation were subjected to histologic observation. Nasal length and nasal tip projection were measured after revision rhinoplasty and the ratio was calculated to evaluate the nasal morphology; patient satisfaction was evaluated using the Likert 5-grade scale. Results: Patients were followed up 12-48 months (mean, 18 months). Inflammation was controlled in 3 patients with infections caused by buried thread nasal augmentation. The remaining 26 patients had satisfactory results immediately after revision rhinoplasty. Before revision rhinoplasty and at 7 days and 6 months after revision rhinoplasty, the nasal length was (4.11±0.34), (4.36±0.25), and (4.33±0.22) cm, respectively; the nasal tip projection was (2.34±0.25), (2.81±0.18), and (2.76±0.15) cm, respectively; and the nasal tip projection/nasal length ratio was 0.57±0.08, 0.65±0.05, and 0.64±0.04, respectively. There were significant differences in the nasal length and the nasal tip projection between time points ( P<0.05). There was a significant difference in the nasal tip projection/nasal length ratio between pre- and post-operation ( P<0.05), but there was no significant difference between 7 days and 6 months after operation ( P>0.05). The Likert score for satisfaction ranged from 1.5 to 5.0 (mean, 4.05). During follow-up period of 26 patients, no nasal prosthesis was exposed, and the shape of the nose was stable, and the nasal skin of 5 patients with exposed threads could be seen with different degrees of scarring; there was no infection, cartilage resorption, and no cartilage deformation, displacement, or exposure. Histological observation showed that absorbable threads were not only absorbed after implantation, but also with the prolongation of time, the inflammatory changes in the surrounding tissues caused by decomposition and absorption of the threads showed a gradual aggravation of the first, the heaviest inflammatory reaction in 6 to 12 months, and then gradually reduce the trend. Conclusion: After implantation of the absorbable thread into the subcutaneous tissue of the nasal dorsum, the nature of the thread is different from the body's own tissue, which will affect the soft tissue compliance of the nasal dorsum. The degradation and absorption of the thread will stimulate the infiltration of inflammatory cells and the proliferation of fibroblasts in the surrounding tissue and then form scar tissue, which will affect the design and effect of revision rhinoplasty.

[Facial nerve-sublingual nerve parallel bridge anastomosis for facial nerve injury caused by closed temporal bone fractures].

Objective: To investigate the effectiveness of facial nerve-sublingual nerve parallel bridge anastomosis for facial nerve injury resulting from closed temporal bone fractures. Methods: Between January 2017 and December 2019, 9 patients with facial nerve injury resulting from closed temporal bone fracture caused by head and face trauma were treated. Among them, 5 patients were treated with facial nerve-sublingual nerve parallel bridge anastomosis (operation group), and 4 patients were treated with neurotrophic drugs combined with rehabilitation exercise (conservative group). There was no significant difference in gender, age, side, cause of injury, duration of facial nerve injury before surgery, House-brackmann grading (hereinafter referred to as HB grading) of facial nerve injury, and other general information between 2 groups ( P>0.05). HB grading was used to evaluate the improvement of facial nerve function before and after treatment. At the same time, facial nerve neuroelectrophysiological test was performed to evaluate the electrical activity of facial muscles before and after treatment. Tongue function, atrophy, and tongue deviation were evaluated after nerve anastomosis according to the tongue function scale proposed by Martins et al. Results: Patients in both groups were followed up 12-30 months, with an average of 25 months. None of the 5 patients in the operation group showed symptoms such as tongue muscle atrophy, tongue extension deviation, hypoglossal nerve dysfunction (mainly including slurred speech, choking with water), postoperative infection, bleeding, lower limb muscle atrophy or lower limb motor dysfunction after sural nerve injury. Postoperative skin sensory disturbance in lateral malleolus area was found, but gradually recovered to normal. During the follow-up, facial nerve and sublingual motor neurons were innervated to paralyzed facial muscle in the operation group. At last follow-up, the HB grading of 5 patients in the operation group improved from preoperative grade Ⅴ in 2 cases, grade Ⅵ in 3 cases to grade Ⅱ in 3 cases, grade Ⅲ in 1 case, and grade Ⅳ in 1 case. And in the conservative group, there were 1 patient with grade Ⅴ and 3 patients with grade Ⅵ before operation, facial asymmetry continued during follow-up, and only 2 patients improved from grade Ⅵ to grade Ⅴ at last follow-up. There was significant difference in prognosis HB grading between the two groups ( t=5.693, P=0.001). In the operation group, the amplitude and frequency of F wave were gradually improved, and obvious action potential could be collected when the facial muscle was vigorously contracted. On the contrary, there was no significant difference in neuroelectrophysiological results before and after treatment in the conservative group. Conclusion: Facial nerve-sublingual nerve parallel bridge anastomosis can effectively retain the integrity of the facial nerve, while introducing the double innervation of the sublingual nerve opposite nerve, which is suitable for the treatment of severe incomplete facial nerve injury caused by closed fracture.

GPBAR1 is associated with asynchronous bone metastasis and poor prognosis of hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death in China. Asynchronous metastasis is the main reason for HCC recurrence, but the current assessment of HCC metastasis and prognosis is far from clinically satisfactory. Materials: In our study, we investigated the expression of G-protein-coupled bile acid receptor (GPBAR1) in HCC tissues and tumor-adjacent tissues by qRT-PCR and immunohistochemistry. The associations between GPBAR1 expression, clinicopathological factors, and asynchronous metastases were assessed by the Chi-square test. The overall survival curves of different variables were plotted with the Kaplan-Meier method, and the statistical significance between different subgroups was analyzed with the log-rank test. The independent prognostic factors were identified by the Cox regression hazard model. Results: GPBAR1 was more highly expressed in HCC tissues than in tumor-adjacent tissues. GPBAR1 expression in HCC was significantly higher than that in liver cirrhosis, followed by normal liver tissues. GPBAR1 was significantly associated with poor prognosis in HCC and can be regarded as an independent prognostic biomarker. Interestingly, GPBAR1 expression in HCC was significantly correlated with asynchronous metastasis to the bone but not to the liver or lung. Conclusions: GPBAR1 was found to be an independent, unfavorable prognostic factor of HCC, as well as an indicator of asynchronous bone metastasis but not liver or lung metastases. Our results could provide a new aspect for HCC metastasis studies and help identify high-risk HCC patients, which helps ameliorate the prognostic assessment of HCC.

Comprehensive Analysis of E3 Ubiquitin Ligases Reveals Ring Finger Protein 223 as a Novel Oncogene Activated by KLF4 in Pancreatic Cancer.

Pancreatic cancer is one of the major malignancies and causes of mortality worldwide. E3 ubiquitin-protein ligases transfer activated ubiquitin from ubiquitin-conjugating enzymes to protein substrates and confer substrate specificity in cancer. In this study, we first downloaded data from The Cancer Genome Atlas pancreatic adenocarcinoma dataset, acquired all 27 differentially expressed genes (DEGs), and identified genomic alterations. Then, the prognostic significance of DEGs was analyzed, and eight DEGs (MECOM, CBLC, MARCHF4, RNF166, TRIM46, LONRF3, RNF39, and RNF223) and two clinical parameters (pathological N stage and T stage) exhibited prognostic significance. RNF223 showed independent significance as an unfavorable prognostic marker and was chosen for subsequent analysis. Next, the function of RNF223 in the pancreatic cancer cell lines ASPC-1 and PANC-1 was investigated, and RNF223 silencing promoted pancreatic cancer growth and migration. To explore the potential targets and pathways of RNF223 in pancreatic cancer, quantitative proteomics was applied to analyze differentially expressed proteins, and metabolism-related pathways were primarily enriched. Finally, the reason for the elevated expression of RNF223 was analyzed, and KLF4 was shown to contribute to the increased expression of RNF233. In conclusion, this study comprehensively analyzed the clinical significance of E3 ligases. Functional assays revealed that RNF223 promotes cancer by regulating cell metabolism. Finally, the elevated expression of RNF223 was attributed to KLF4-mediated transcriptional activation. This study broadens our knowledge regarding E3 ubiquitin ligases and signal transduction and provides novel markers and therapeutic targets in pancreatic cancer.

High-throughput three-dimensional spheroid tumor model using a novel stamp-like tool.

Spheroid culture is a widely used three-dimensional culture technology that simulates the three-dimensional structure of tumors in vivo and has been considered a good model for tumor research. However, current commercialized spheroid culture tools have the shortcomings of high cost or relatively poor spheroid-forming results for some special cells. To solve such problems, we designed a 3D printed, reusable, stamp-like resin mold that could shape microstructures for spheroid culture of tumor cells on the surface of agarose substrate in a 96-well plate. We applied this homemade three-dimensional culture tool in spheroid formation for hepatocellular carcinoma cells. The experimental data show that the effect of spheroid culture on four hepatocellular carcinoma cell lines in our homemade spheroid culture plate is better than that of the commercialized ultralow attachment spheroid culture plate, and compared to two-dimensional culture, three-dimensional culture improves cell functions. In addition, the drug-sensitive test based on patient-derived hepatocellular carcinoma cells showed a different pattern between spheroid and two-dimensional cultures. In conclusion, our spheroid culture tool is characterized by its low cost, reusability, low cell consumption, convenience in medium exchange, and good effect of spheroid formation, suggesting that this technique could be widely used in individual treatment and high-throughput drug screening.

Comparison of in vivo adipogenic capabilities of two different extracellular matrix microparticle scaffolds.

BACKGROUND: The extracellular matrix is an essential microenvironment for cell survival activity. The adipose tissue extract microparticle scaffolds from human adipose tissue and small intestine submucosa microparticle scaffolds from porcine jejunum were prepared. Their effects on the adipogenic capabilities of human adipose-derived stem cells were compared in vivo. METHODS: A combination of physical and chemical methods was used to decellularize human fat and porcine jejunum. Expression of CD molecules on the adipose-derived stem cell surface was determined by flow cytometry. The stem cells were then cultured with the scaffold materials in vitro. The cell-scaffold complexes were implanted subcutaneously into nude mice, and samples were collected 4 and 8 weeks later. The adipogenic differentiation capabilities of adipose-derived stem cells were studied by histologic methods and real-time polymerase chain reaction. RESULTS: The authors observed high expression of CD90 and CD44; no expression of CD34, CD45, CD31, or CD106; and weak positive expression of CD49d on the extracted cells, which indicates that the cells were adipose-derived stem cells. The main constituent of the decellularized adipose tissue extract and small intestine submucosa microparticles was collagenous fiber, and the cells proliferated faster on the adipose tissue extract than on small intestine submucosa. Formation of adipocytes in the adipose tissue extract group was closer to that of normal human fat tissue compared with that of the small intestine submucosa group. CONCLUSIONS: Extracellular matrix microparticle scaffolds could promote proliferation, adhesion, and adipogenic differentiation of adipose-derived stem cells. The role of the adipose tissue extract microparticle scaffold in promoting adipogenesis was stronger and more suitable as a vector in fatty tissue engineering.