The GE296_RS03820 and GE296_RS03830 genes are involved in capsular polysaccharide biosynthesis in Riemerella anatipestifer.

Riemerella anatipestifer is a pathogenic bacterium that causes duck serositis and meningitis, leading to significant harm to the duck industry. To escape from the host immune system, the meningitis-causing bacteria must survive and multiply in the bloodstream, relying on specific virulence factors such as capsules. Therefore, it is essential to study the genes involved in capsule biosynthesis in R. anatipestifer. In this study, we successfully constructed gene deletion mutants Δ3820 and Δ3830, targeting the GE296_RS03820 and GE296_RS03830 genes, respectively, using the RA-LZ01 strain as the parental strain. The growth kinetics analysis revealed that these two genes contribute to bacterial growth. Transmission and scanning electron microscopy (TEM and SEM) and silver staining showed that Δ3820 and Δ3830 produced the altered capsules and compounds of capsular polysaccharides (CPSs). Serum resistance test showed the mutants also exhibited reduced C3b deposition and decreased resistance serum killing. In vivo, Δ3820 and Δ3830 exhibited markedly declining capacity to cross the blood-brain barrier, compared to RA-LZ01. These findings indicate that the GE296_RS03820 and GE296_RS03830 genes are involved in CPSs biosynthesis and play a key role in the pathogenicity of R. anatipestifer. Furthermore, Δ3820 and Δ3830 mutants presented a tendency toward higher survival rates from RA-LZ01 challenge in vivo. Additionally, sera from ducklings immunized with the mutants showed cross-immunoreactivity with different serotypes of R. anatipestifer, including 1, 2, 7 and 10. Western blot and SDS-PAGE assays revealed that the altered CPSs of Δ3820 and Δ3830 resulted in the exposure of some conserved proteins playing the key role in the cross-immunoreactivity. Our study clearly demonstrated that the GE296_RS03820 and GE296_RS03830 genes are involved in CPS biosynthesis in R. anatipestifer and the capsule is a target for attenuation in vaccine development.

A Comparative Study on Platelet-Rich Plasma From Elderly Individuals and Young Adults to Treat Pressure Ulcers in Mice.

OBJECTIVE: The aim of the present study was to compare the therapeutic effects of activated platelet-rich plasma (PRP) prepared from elderly individuals and young adults to treat pressure ulcers (PUs), and to accumulate a theoretical basis for allogeneic PRP treatment of PUs in elderly patients. MATERIALS AND METHODS: Whole blood was extracted from elderly individuals aged >65 y and young adult volunteers for PRP preparation, and platelet concentrations in whole blood and PRP were compared. Growth factors released from activated PRP were assayed using the enzyme-linked immunosorbent assay. C57BL/6 mice were divided into three groups: the control saline, elderly-PRP (Group A), and young adult-PRP (Group B). Ischemia-reperfusion injury-induced PUs were established on the backs of mice. PUs were photographed on days 0, 5, and 10 to assess their sizes. Specimens were collected on day 10 and subjected to hematoxylin and eosin and Masson's staining. Immunohistochemical staining for CD31 was conducted to evaluate vascular formation, and cell invasion was assessed using a Transwell assay. The action of PRP on transforming growth factor-beta (TGF-ß)-dependent fibroblast activity and epithelial-mesenchymal transition was analyzed using immunofluorescence and Western blotting in vitro. RESULTS: The platelet concentrations in whole blood and PRP of young adults were significantly higher than that in elderly individuals. The two PRP treatment groups had similar platelet enrichment coefficients of PRP. After activation, PRP from young adults produced significantly higher levels of platelet-derived growth factor, TGF-ß, and vascular endothelial growth factor than PRP from elderly individuals (P < 0.05). The concentrations of platelet-derived growth factor, TGF-ß, and vascular endothelial growth factor were positively correlated with the platelet concentrations in whole blood and PRP. The effects of PRP in regulating the expressions of TGF-ß, α-smooth muscle actin, vimentin, and E-cadherin were observed in vivo and in vitro. The two PRP treatment groups exhibited better wound healing than the control group, as evidenced by more re-epithelialization, higher collagen content, skin fibrosis, and more blood vessel formation over time. Group B exhibited better wound healing than Group A (P < 0.05). CONCLUSION: PRP exhibits potent wound healing ability in PU therapy, and PRP from young adults is seemingly superior to that from elderly individuals because of a higher concentration of platelets and increased production of growth factors.

Recombinant human growth hormone promotes wound angiogenesis in burned mice through the ERK signaling pathway.

Burns are the most severe type of trauma, and the resulting ischemia and hypoxia damage can promote the dysfunction and even failure of tissues and organs throughout the body, endangering patients' life safety. Recombinant human growth hormone (rhGH) has the functions of promoting protein synthesis to reverse negative nitrogen balance, accelerating wound healing, and improving immune function, which is widely used in the treatment of burns. However, the exact mechanism and pathway of rhGH's action is not yet fully understood. In this study, we observed the wound repair effect of recombinant human growth hormone (rhGH) on burned mice and further analyzed the mechanism of action, which can provide more comprehensive reference opinions for clinical practice. First, by establishing a burn mouse model and and intervening with different doses of rhGH, we found that the wound healing capacity of mice was significantly enhanced and the inflammatory and oxidative stress responses were obviously alleviated, confirming the excellent promotion of wound repair and anti-inflammatory and antioxidant effects of rhGH. Subsequently, we found that the expression of p-ERK1/2/ERK1/2, EGF, TGF-ß, and VEGF proteins was elevated in the traumatic tissues of mice after rhGH intervention, suggesting that the pathway of action of rhGH might be related to the activation of ERK pathway to promote the regeneration of traumatic capillaries.

Mentalis Muscle Reconstruction: An Innovative Technique.

BACKGROUND: The mentalis muscle is a significant component of the lower lip; its injury could impair appearance and function. This study presents a surgical strategy for treating mentalis muscle rupture to restore muscle tension and improve function. METHODS: Medical records and photographs of 2 patients with mentalis muscle rupture were reviewed. After physical examinations, 3-dimensional computed topographies were conducted to evaluate chin appearance further. The surgical strategy was designed according to individual malformations and requests. RESULTS: Both patients injured their mentalis muscles in childhood and presented to our clinic with a concave deformity at the center of the chin and impaired mentalis muscle function. The surgical procedure involved the reconstruction of the mentalis muscle by reassembling the muscle flaps and releasing the scarred soft tissue. Both patients were satisfied with the outcome. CONCLUSIONS: Mentalis muscle rupture requires surgical correction. The study proposes an innovative approach that enables patients to achieve a more harmonious appearance and improve function.

Construction of a Lentiviral Vector for Fgfr2 Overexpression and its Impact on the Biological Behavior of Cranial Suture Mesenchymal Stem Cells.

OBJECTIVE: Suture mesenchymal stem cells (SuSCs), possessing self-renewal and multilineage differentiation abilities, play a crucial role in cranial bone growth. However, the impact of the disease-causing fibroblast growth factor receptor 2 (FGFR2) mutation on SuSCs in Crouzon syndrome has not been explored. This study aims to employ a lentivirus to overexpress Fgfr2 and investigate its role in the pathogenesis of Crouzon syndrome. METHODS: Starting with the prevalent FGFR2 mutation site in patients with Crouzon syndrome, a lentiviral vector carrying the Fgfr2.C361Y mutation was developed and transfected into SuSCs, with a determined multiplicity of infection values. The experimental group, SuSCs+Fgfr2.C361Y, was compared with the empty vector and normal SuSC groups. Cell proliferation, cycle, apoptosis, and osteogenic functionality were assessed using CCK-8 assays, flow cytometry, ALP activity assays, and real-time quantitative polymerase chain reaction. RESULTS: The lentiviral vector effectively infected SuSCs, leading to heightened Fgfr2 expression, with optimal multiplicity of infection values of 80. The experimental group demonstrated decreased proliferation activity and a higher apoptosis rate compared with controls (P < 0.05). After osteogenic induction, the experimental group showed significantly higher ALP activity than controls (P < 0.05). Real-time quantitative polymerase chain reaction indicated lower mRNA expression levels of Gli1, Axin2, Pcna, Cdk2, and Bcl-2 in the experimental group than controls, whereas Bax, Runx2, and Bmp-2 showed higher expression (P < 0.05). CONCLUSION: This study constructed a lentivirus vector to upregulate Fgfr2 expression in SuSCs, suppressing stem cell stemness by inhibiting proliferation, promoting apoptosis, and accelerating premature osteogenic differentiation, resulting in premature suture closure. These findings establish the groundwork for further understanding the pathogenesis of Crouzon syndrome.

Efficacy and Safety of PL-5 (Peceleganan) Spray for Wound Infections: A Phase IIb Randomized Clinical Trial.

OBJECTIVE: To assess the safety and efficacy of antimicrobial peptide PL-5 (Peceleganan) spray in the treatment of wound infections. BACKGROUND: Antimicrobial peptide PL-5 spray is a novel topical antimicrobial agent. METHODS: We conducted a multicenter, open-label, randomized, controlled phase IIb clinical trial to evaluate the efficacy and safety of PL-5 spray, as compared with silver sulfadiazine, in patients with skin wound infections. The primary efficacy outcome was the clinical efficacy rate on the first day after ending the treatment (D8). The secondary efficacy outcome was the clinical efficacy rate on the fifth day posttreatment (D5), the bacteria clearance rate, and the overall efficacy rate at the mentioned 2 time points. The safety outcomes included adverse reactions and pharmacokinetic analysis posttreatment. RESULTS: A total of 220 patients from 27 hospitals in China were randomly assigned to 4 groups. On D8, the efficacy rate was 100.0%, 96.7%, 96.7% for the 1‰ PL-5, 2‰ PL-5, 4‰ PL-5 groups, respectively, as compared with 87.5% for the control group. The efficacy rate among the 4 groups was significantly different ( P <0.05). On D5, the efficacy rate was 100.0%, 93.4%, 98.3% for the 1‰ PL-5, 2‰ PL-5, 4‰ PL-5 groups, respectively, as compared with 82.5% for the control group. The efficacy rate among the 4 groups was significantly different ( P <0.05). The blood concentration of PL-5 was not detectable in pharmacokinetic analysis. No severe adverse event related to the application of PL-5 was reported. CONCLUSIONS: Antimicrobial peptide PL-5 spray is safe and effective for the treatment of skin wound infections. TRIAL REGISTRATION: ChiCTR2000033334.

Amide proton transfer-weighted MRI features of acute ischemic stroke subtypes.

Stroke is a highly heterogeneous disorder with distinct subtypes, and the stroke subtype influences the outcome. Amide proton transfer-weighted (APTW) MRI has been demonstrated to be promising in stroke patients, but the image characteristics of stroke subtypes have not been sufficiently investigated. The purpose of this study was to investigate the APTW MRI features of different subtypes of acute ischemic stroke (AIS). Ninety-two AIS patients presenting within 96 h of symptom onset were enrolled and examined with a 3.0-T MRI system. Patients were grouped into four subtypes: lacunar circulation infarcts (LACI, n = 33); total anterior circulation infarcts (TACI, n = 9); partial anterior circulation infarcts (PACI, n = 28); and posterior circulation infarcts (POCI, n = 22). APTW values in the lesion (APTWlesion ) and the contralateral normal-appearing region (APTWcontral ) were measured. The change in APTW values between the acute ischemic lesion and the contralateral normal-appearing region (APTWles-con ) was calculated. A two-sample t-test, one-way ANOVA, and the Chi-square method were used. There were significant differences between APTWlesion and APTWcontral in the three categories of nonlacunar strokes (TACI, PACI, and POCI, all p < 0.01), but not for lacunar strokes (LACI, p = 0.080). TACI patients had the lowest APTWlesion and APTWles-con in all groups (p < 0.05). In the POCI group, patients with supratentorial infarcts showed significant differences between APTWlesion and APTWcontral (p = 0.001), while the differences were not significant for infratentorial infarcts (p = 0.135). Our results suggest that the APT effect was heterogeneous in different stroke subtypes, and that APTW MRI gave an excellent performence in depicting nonlacunar AIS in supratentorial locations.

Circulating metabolites as potential biomarkers for the early detection and prognosis surveillance of gastrointestinal cancers.

BACKGROUND AND AIMS: Two of the most lethal gastrointestinal (GI) cancers, gastric cancer (GC) and colon cancer (CC), are ranked in the top five cancers that cause deaths worldwide. Most GI cancer deaths can be reduced by earlier detection and more appropriate medical treatment. Unlike the current "gold standard" techniques, non-invasive and highly sensitive screening tests are required for GI cancer diagnosis. Here, we explored the potential of metabolomics for GI cancer detection and the classification of tissue-of-origin, and even the prognosis management. METHODS: Plasma samples from 37 gastric cancer (GC), 17 colon cancer (CC), and 27 non-cancer (NC) patients were prepared for metabolomics and lipidomics analysis by three MS-based platforms. Univariate, multivariate, and clustering analyses were used for selecting significant metabolic features. ROC curve analysis was based on a series of different binary classifications as well as the true-positive rate (sensitivity) and the false-positive rate (1-specificity). RESULTS: GI cancers exhibited obvious metabolic perturbation compared with benign diseases. The differentiated metabolites of gastric cancer (GC) and colon cancer (CC) were targeted to same pathways but with different degrees of cellular metabolism reprogramming. The cancer-specific metabolites distinguished the malignant and benign, and classified the cancer types. We also applied this test to before- and after-surgery samples, wherein surgical resection significantly altered the blood-metabolic patterns. There were 15 metabolites significantly altered in GC and CC patients who underwent surgical treatment, and partly returned to normal conditions. CONCLUSION: Blood-based metabolomics analysis is an efficient strategy for GI cancer screening, especially for malignant and benign diagnoses. The cancer-specific metabolic patterns process the potential for classifying tissue-of-origin in multi-cancer screening. Besides, the circulating metabolites for prognosis management of GI cancer is a promising area of research.

Early globalized industrial chain revealed by residual submicron pigment particles in Chinese imperial blue-and-white porcelains.

The success of early Chinese blue-and-white porcelains relied heavily on imported cobalt pigment from the West. In contrast to art-historical concept, which contains both typological evidence and literature records, it is assumed that imported Sumali blue was completely superseded by domestic Chinese asbolane ore based on the analytical results of the Fe/Mn ratio in imperial productions from the Xuande reign (1426 to 1435 CE) onward. Using a focused ion beam transmission electron microscopy technique to reassess this hotly debated question, we have identified two classes of residual submicron pigment particles in the blue glaze with diagnostic differences in morphology, chemical composition, and distribution behavior. Compared with the microstructural features of the blue-and-white porcelains of the Yuan and Qing dynasties, we show that a mixture of imported and domestic cobalt pigments was used for aesthetic reasons, indicating that the overseas supply chain of imported pigment remained consistent and adequate even though the authorities had terminated official trade and tributary activities after the death of Admiral Zheng He. This discovery further suggests that the globalized trading network and cross-regional industrial chain had been extensively established in the 15th century. Moreover, we provide analytical evidence against the fundamental assumption of the current Fe/Mn provenancing criteria, implying that the failures of previous chemical analyses can be attributed to elemental differentiation between the silicate glaze and the arsenic pigment. We propose an innovative method for directly assessing original mineralogic information from submicron residual pigment particles that provides a more reliable way to trace cobalt circulation and holds great promise for provenance studies.

Long noncoding RNA CASC9 promotes pancreatic cancer progression by acting as a ceRNA of miR-497-5p to upregulate expression of CCND1.

BACKGROUND: Pancreatic cancer (PC) is an aggressive malignancy with poor prognosis. Accumulating studies have showed that long non-coding RNA (lncRNA) is a crucial regulator in various tumorigenesis and progression including PC. This research aims to explore the roles and molecular mechanism of lncRNA cancer susceptibility candidate 9 (CASC9) in PC. METHODS: The expression levels of lncRNA CASC9 and miR-497-5p were evaluated in PC tissues and paired adjacent healthy tissues by quantitative real-time PCR. PC cell lines were transfected with lentivirus targeting lncRNA CASC9, and cells proliferation, migration and invasion tests were conducted. Dual luciferase reporter assays were also carried out to explore the relationship between lncRNA CASC9, miR-497-5p and Cyclin D1 (CCND1). RESULTS: LncRNA CASC9 was significantly up-regulated in PC tissues, while miR-497-5p expression was down-regulated. Down-regulation of lncRNA CASC9 in PC cells can significantly suppress the cell aggressiveness both in vitro and in vivo; moreover, knock-down of miR-497-5p could neutralize this impact. Additionally, the luciferase activity assay has assured that CCND1 was a downstream target of miR-497-5p. CONCLUSION: LncRNA CASC9 can promote the PC progression by modulating miR-497-5p/CCND1 axis, which is potential target for PC treatment.

Adenoviral Vector for Enhanced Prostate Cancer Specific Transferrin Conjugated Drug Targeted Therapy.

Prostate cancer (PCa) is one of the leading causes of death for men worldwide. Unlike some other types of cancer, there is a lack of targeted therapy for prostate cancer patients that can kill cancer cells but do much less damage to the normal tissue. In this paper, we report on an adenoviral vector enhanced prostate cancer specific transferrin conjugated drug targeted therapy. In particular, a functional PCa-specific gene probe is introduced to drive and up-regulate the transferrin receptor expression on the PCa via adenoviral vector. As a result, significantly enhanced accumulation of nanoscale transferrin-doxorubicin (Tf-DOX) protein drug conjugates and concomitant notably elevated PCa tumor inhibition are observed. This conceptual strategy provides the proof-of-concept for the targeted therapy of PCa that is highly desired but not yet developed.

Redefining Mandibular Sub-units: The "ABC" Classification and Surgical Strategies for Lower Face Contouring.

BACKGROUND: In east Asia, lower face contouring surgeries including reduction mandibuloplasty and genioplasty are the most popular aesthetic craniofacial surgeries. Conventional selection of surgical strategies mainly relied on the visual judgment of the mandibular angle, without overall assessment of the mandibular sub-units. Furthermore, only a few studies offered quantitative assessment of the mandibular shape. METHODS: From 2010 to 2021, 1241 patients diagnosed with square faces and received customized lower face contouring surgeries by the senior author were reviewed and analyzed to propose an "ABC" classification system for facilitating surgical planning. RESULTS: Among them, 998 (80.42%) received bilateral mandible reshaping, 155 (12.49%) underwent bilateral mandible reshaping combined with genioplasty, and 88 (7.09%) received asymmetric mandible reshaping. A modified classification system composed of three critical parameters (height, morphology/thickness, divergence) in three aesthetic zones (mandibular angle, mandibular body, chin) was proposed based on quantitative summarization of the CT database and the senior author's 12-year experience. The way to facilitate surgical planning with this classification was demonstrated. CONCLUSIONS: This modified classification system ushered a decision-making process that prioritized several critical measurements and proposed an operative planning form. Meanwhile, it can also be cooperated into the three-dimensional virtual surgical plan. 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 http://www.springer.com/00266 .

Safety and efficacy of laparoscopic common bile duct exploration for the patients with difficult biliary stones: 8 years of experiences at a single institution and literature review.

BACKGROUND: Laparoscopic common bile duct exploration (LCBDE) has gained wide popularity for the treatment of choledocholithiasis. However, it remains unclear whether LCBDE is a better alternative option for the patients with difficult biliary stones. Thus, the aim of the present study was to explore the safety and efficacy of LCBDE for these patients by retrospectively analyzing our data and combing with literature review. METHODS: Between September 2011 and February 2019, 1064 consecutive patients who underwent LCBDE at Shanghai Tenth People's Hospital were reviewed. The clinical data of patients with difficult biliary stones were selected and retrospectively analyzed. RESULTS: Of these patients, 334 cases were confirmed with difficult biliary stones, and the overall complete stone clearance rate was 98.8% (330/334). 34 cases (10.2%) were performed with laser lithotripsy. A total of 296 patients (88.6%) underwent primary closure of common bile duct, and T-tube drainage was indwelled in 38 patients (11.4%). No bile duct injury, bleeding, perforation and surgery-related deaths were observed. The overall morbidity rate was 6.6%. 16 cases (4.8%) occurred in bile leakage with primary closure procedure, and all of them were managed successfully with conservative therapy. The median follow-up period was 9 months with stone recurrence occurring in 9 patients (2.7%). There was no evidence of bile duct stricture in all cases. CONCLUSIONS: The current study suggests that LCBED is a considerable safe and effective option for the patients with difficult biliary stones. A randomized clinical trial is needed to further evaluate the benefit of LCBDE in this subgroup.

Surgical Result and Identification of FGFR2 Variants Using Whole-Exome Sequencing in a Chinese Family With Crouzon Syndrome.

ABSTRACT: Crouzon syndrome is considered as one of the most common craniosynostosis syndromes with a prevalence of 1 in 65,000 individuals, and has a close relationship with variants in fibroblast growth factor receptor 2. Here the authors described a Crouzon syndrome case, which was asked for surgery treatment for the symptom of multisuture craniosynostosis. Mild midfacial retrusion, larger head circumference, proptosis, pseudo-prognathism, and dental malposition could also be found obviously. Then fronto-orbital advancement and cranial cavity expansion were performed to the child. After whole-exome sequencing (WES) and Sanger sequencing, gene variants in the exons 2 and 3 of FGFR2 were detected. And protein tyrosine 105 replaced by cysteine in the extracellular region of FGFR2 was also detected. After operation, she presented a satisfactory anterior plagiocephaly and scaphocephaly correction, and the result was satisfied by surgeons and her parents. Variants detected using WES have further research prospect.

The Effect of Yes-Associated Protein on the Interaction Between the MEK/Extracellular Signal-Regulated Kinase and Hippo Pathways in Osteoblasts Co-Cultured With Fibroblast Growth Factor Receptor 2-Mutated Dura Cells.

BACKGROUND: C342Y (Cys342Tyr) point mutation of FGFR2 (fibroblast growth factor receptor 2) is closely associated with the pathogenesis of Crouzon syndrome. The dura mater plays an important role in mediating the closure of cranial sutures. However, the underlying mechanisms of these pathological processes have been rarely investigated. in this study, the authors analyzed the effects of dura cells with FGFR2 mutations on the biological function of osteoblasts. METHODS: Dura cells and cranial osteoblasts from C57BL/6 mice were extracted and cultured. C342Y-FGFR2 mutant constructs were established via lentivirus and applied to infect dura cells. A co-cultured trans-well system with dura cells and osteoblasts was established. Three experimental groups were set up: oste group, Oste + Dura-vector group, and Oste + Dura-C342Y group. The expression levels of key factors in MEK (Mitogen-activated protein kinase kinase, MAPKK)/extracellular signal-regulated kinase (ERK) and Hippo pathway were detected by western blot and RT-qPCR (Real Time Quantitative PCR). Finally, a rescue experiment was carried out with small interference RUA. RESULTS: The proliferation level of osteoblasts in Oste + Dura- C342Y group was significantly up-regulated. Our studies indicated that the activation of MEK/ERK pathway in Oste + Dura-C342Y group could inhibit the Hippo pathway, lead to down-regulation of large tumor suppressor 1 and promote the activation and nuclear localization of yes-associated protein, and the results of rescue experiments showed a reverse expression trend, further confirming the effects of C342Y-FGFR2 mutation in dura cells on osteoblasts and its potential mechanism. CONCLUSIONS: This study suggested that the C342Y-FGFR2 mutation in dura cells could promote osteoblastic proliferation, and shown the crosstalk between MEK/ERK and Hippo pathways. As the regulatory machinery center, yes-associated protein might play a bridging role in these pathways, and might influence the pathogenesis of craniosynostosis by activating downstream transcriptional factors.

Mesenchymal stromal cell therapy for pancreatitis: Progress and challenges.

Pancreatitis is a common gastrointestinal disease with no effective therapeutic options, particularly for cases of severe acute and chronic pancreatitis (CP). Mesenchymal stromal cells (MSCs) are multipotent cells with diverse biological properties, including directional migration, paracrine, immunosuppressive, and antiinflammatory effects, which are considered an ideal candidate cell type for repairing tissue damage caused by various pathogenies. Several researchers have reported significant therapeutic efficacy of MSCs in animal models of acute and CP. However, the specific underlying mechanisms are yet to be clarified and clinical application of MSCs as pancreatitis therapy has rarely been reported. This review mainly focuses on the potential and challenges in clinical application of MSCs for treatment of acute and CP, along with discussion of the underlying molecular mechanisms.

VRK1 promotes proliferation, migration, and invasion of gastric carcinoma cells by activating ß-catenin.

Vaccinia-related kinase 1 (VRK1) is a member of the VRK subfamily belonging to the casein kinase superfamily, and it regulates the proliferation and survival of cells both in normal and malignant tissues. A variety of transcription factors including c-Jun can also be specifically phosphorylated and stimulated by VRK1. However, the regulatory mechanism of VRK1 in gastric carcinoma (GC) remains unclear. This research aimed to determine the function of VRK1 during tumor progression in GC. The mRNA and protein expression of the VRK1 and other genes were evaluated in GC cell lines using real-time RT-PCR and western blotting. Cell proliferation was analyzed using the cell count kit-8 (CCK-8) assay, and cell migration and invasion were monitored by the Transwell assay. The downregulated genes in shVRK1 cells compared with shCtrl were assessed using RNA-seq. The interactions of VRK1 with ß-catenin or c-Jun were detected by co-IP. We found that VRK1 was overexpressed in gastric cancer cells, conversely, knockdown of VRK1 inhibits GC cells' proliferation, migration, and invasion. Moreover, VRK1 might regulate the expression of ß-catenin (CTNNB1) at the transcriptional level by phosphorylating c-Jun, the transcriptional factor of ß-catenin. VRK1 changes the subcellular location and decreases the nuclear aggregation of c-Jun by phosphorylating the Ser243 site. To conclude, VRK1 can affect migration and invasion by regulating the expression of ß-catenin at the transcriptional level in GC cells.

A Simple Dressing Fixation Method Following Harvest of the Ear Cartilage Graft.

ABSTRACT: Auricular cartilage is a common autologous graft material for rhinoplasty. Even though surgical techniques for the harvest of cartilage graft are well established, the management of the postoperative dressing fixation is still limited. Therefore, the authors propose a simple dressing fixation method in which gauze strips instead of gauze block or wet cotton, the suture of tie-over dressing fixed just through the cartilage rather than the full thickness of the auricle, a vaseline gauze interposed between the stitch knot and the skin and no additional pressure dressings postauricularly. No case of hematoma, infection and skin necrosis of the donor site was observed. This simple and reproducible technique provides perfect and homogeneous adhesion of the dressing all over the conchal cartilage while decreasing the risk of postoperative hematoma and discomfort.

Facial Contour Refining after Surgery-First SSRO with Computer-Assisted Design in East Asians.

BACKGROUND: Surgery-first sagittal split ramus osteotomies (SF-SSRO) are an effective treatment for patients with dental malocclusion. However, some patients with mandibular prognathism usually have facial deficiencies which cannot be corrected completely after orthognathic surgery. These are not accepted because the remaining facial contours are in disharmony. METHODS: Twenty-five patients, who were unsatisfied with their appearances after SF-SSRO and orthodontics, were included. The preoperative CTs were used to investigate patients for facial deformity. To achieve a harmonious facial contour, mandible long-curve osteotomy/mandible U-shaped osteotomy, genioplasty or facial autologous fat grafting was selected depending on patients' contour deformities with the assistance of CAD. RESULTS: Among the patients who underwent osteotomy, the gonial angle was improved from 111.16° ± 9 to 111.58° ± 9.06 after SF-SSRO because of distal fragment setback and rotation. After facial refine surgery, the gonial angle was significantly increased to 121.69°±2.41 (p < 0.05). And the mandibular width was decreased from 11.29 cm ± 0.44 to 10.45 cm ± 0.39 (p < 0.05) after mandibular outer plate griding. All patients were shown no signs of infection, massive bleeding, and osteonecrosis in the early stage. After follow-up time, they were all satisfied with their results and most of them recovered from lip numbness. CONCLUSIONS: This study indicated the clinical feasibility of two-stage orthognathic and facial bone contouring surgery for the treatment of dentofacial deformities. Two-stage facial contouring surgery can provide esthetic improvement for more accuracy in refining the facial contour. 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 .

Rapamycin and 3-Methyladenine Influence the Apoptosis, Senescence, and Adipogenesis of Human Adipose-Derived Stem Cells by Promoting and Inhibiting Autophagy: An In Vitro and In Vivo Study.

OBJECTIVE: We aimed to clarify the changes in apoptosis, proliferation, senescence, and adipogenesis after promoting and inhibiting autophagy in adipose-derived stem cells (ADSCs) by rapamycin and 3-methyladenine in vitro and in vivo. METHODS: After rapamycin and 3-methyladenine pretreatment, ADSC autophagy was detected by immunofluorescence for LC3, RT-PCR for ATG genes, and western blotting (WB) for the LC3 II/I and p62 proteins. TUNEL staining, PCR of BAX, and WB of Caspase-3 were preformed to assess ADSC apoptosis. The adipogenesis of ADSCs was evaluated by Oil red O staining and PCR of PPAR-γ. CCK8 assays were conducted to detect proliferation. Senescence was tested by Sa-ß-gal staining and PCR of the P16/ 19/21 genes. Moreover, the mass and volume retention rate were determined, and perilipin and CD31 staining were performed in vivo. RESULTS: Rapamycin and 3-methyladenine pretreatment increased and decreased autophagy of ADSCs, respectively, under normal and oxygen-glucose deprivation conditions. Apoptosis and senescence of ADSCs were decreased, and adipogenesis was increased along with the upregulation of autophagy. However, the proliferation of ADSCs was inhibited after either rapamycin or 3-methyladenine pretreatment. In vivo, the volume and mass retention rate and the angiogenesis of the grafts were also improved after rapamycin pretreatment. CONCLUSIONS: Rapamycin pretreatment reduced apoptosis, delayed senescence, and promoted adipogenesis of ADSCs. These effects were inhibited by 3-methyladenine, indicating that the changes may be mediated by autophagy. Moreover, the survival rate and angiogenesis of the grafts were increased after upregulation of ADSC autophagy in vivo, which may help improve the efficiency of clinical fat transplantation. NO LEVEL ASSIGNED: 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 .