Platelet-Rich Plasma Lysate Enhances the Osteogenic Differentiation of Adipose-Derived Stem Cells.

ABSTRACT: Adipose-derived stem cells (ADSCs) have become an accepted source of cells in bone tissue engineering. This study aimed to investigate whether platelet-rich plasma (PRP) lysate can replace traditional fetal bovine serum as a culture medium with the enhanced proliferation and osteogenic potential of ADSCs. We divided the experiment into 5 groups where the ADSCs were cultured in an osteogenic medium containing 2.5%, 5%, 7.5%, and 10% PRP lysate with 10% fetal bovine serum as the control group. The cell proliferation, alkaline phosphatase (ALP) activity, ALP stain, alizarin red stain, osteocalcin (OCN) protein expression, and osteogenic-specific gene expression were analyzed and compared among these groups. The outcome showed that all PRP lysate-treated groups had good ALP stain and ALP activity performance. Better alizarin red stains were found in the 2.5%, 5%, and 7.5% PRP lysate groups. The 2.5% and 5% PRP lysate groups showed superior results in OCN quantitative polymerase chain reaction, whereas the 5% and 7.5% PRP lysate groups showed higher OCN protein expressions. Early RUNX2 (Runt-related transcription factor 2 () genes were the most expressed in the 5% PRP lysate group, followed by the 2.5% PRP lysate group, and then the 7.5% PRP lysate group. Thus, we concluded that 5% PRP lysate seemed to provide the optimal effect on enhancing the osteogenic potential of ADSCs. Platelet-rich plasma lysate-treated ADSCs were considered to be a good cell source for application in treating nonunion or bone defects in the future.

LncRNA CARMN suppresses EMT through inhibiting transcription of MMP2 activated by DHX9 in breast cancer.

Long non-coding RNAs (lncRNAs) have been shown to drive cancer progression. However, the function of lncRNAs and the underlying mechanism in early-stage breast cancer(BC) have rarely been investigated. Datasets of pre-invasive ductal carcinoma in situ (DCIS), invasive ductal BC (IDC) and normal breast tissue from TCGA and GEO databases were used to conduct bioinformatics analysis. LncRNA CARMN was identified as a tumor suppressor in early-stage BC and related to a better prognosis. CARMN over-expression inhibited MMP2 mediated migration and EMT in BC. Further analysis showed that CARMN was located in the nucleus and functioned as an enhancer RNA (eRNA) in mammary epithelial cell. Mechanically, CARMN binding protein DHX9 was identified by RNA pull-down and mass spectrometry (MS) assays and it also bound to the MMP2 promoter to activate its transcription. As a decoy, CARMN competitively bound to DHX9 and blocked MMP2 transcriptional activation, thereby inhibiting metastasis and EMT of BC cells. These findings reveal the important role of CARMN as a tumor suppressor in the metastasis and a potential biomarker for progression in early-stage BC.

Genomic alterations driving precancerous to cancerous lesions in esophageal cancer development.

Esophageal squamous cell carcinoma (ESCC) develops through a series of increasingly abnormal precancerous lesions. Previous studies have revealed the striking differences between normal esophageal epithelium and ESCC in copy number alterations (CNAs) and mutations in genes driving clonal expansion. However, due to limited data on early precancerous lesions, the timing of these transitions and which among them are prerequisites for malignant transformation remained unclear. Here, we analyze 1,275 micro-biopsies from normal esophagus, early and late precancerous lesions, and esophageal cancers to decipher the genomic alterations at each stage. We show that the frequency of TP53 biallelic inactivation increases dramatically in early precancerous lesion stage while CNAs and APOBEC mutagenesis substantially increase at late stages. TP53 biallelic loss is the prerequisite for the development of CNAs of genes in cell cycle, DNA repair, and apoptosis pathways, suggesting it might be one of the earliest steps initiating malignant transformation.

Transcriptionally regulated miR-26a-5p may act as BRCAness in Triple-Negative Breast Cancer.

BACKGROUND: DNA damage and DNA damage repair (DDR) are important therapeutic targets for triple-negative breast cancer (TNBC), a subtype with limited chemotherapy efficiency and poor outcome. However, the role of microRNAs in the therapy is emerging. In this study, we explored whether miR-26a-5p could act as BRCAness and enhance chemotherapy sensitivity in TNBC. METHODS: Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-26a-5p in breast cancer tissues and cell lines. CCK-8 was used to measure drug sensitivity in concentration gradient and time gradient. Comet assay was used to detect DNA damage. Flow cytometry was performed to examine apoptosis. Moreover, we used western blot and immunofluorescence to detect biomarkers. Luciferase reporter assay was performed to verify the combination of miR-26a-5p and 3'UTR of target gene. Hormone deprivation and stimulation assay were used to validate the effect of hormone receptors on the expression of miR-26a-5p. Chromatin immunoprecipitation (ChIP) assays were used to verify the binding sites of ER-a or PR with the promoter of miR-26a-5p. Animal experiments were performed to the effect of miR-26a-5p on Cisplatin treatment. RESULTS: The expression of miR-26a-5p was significantly downregulated in TNBC. Overexpressing miR-26a-5p enhanced the Cisplatin-induced DNA damage and following apoptosis. Interestingly, miR-26a-5p promoted the expression of Fas without Cisplatin stimulating. It suggested that miR-26a-5p provided a hypersensitivity state of death receptor apoptosis and promoted the Cisplatin sensitivity of TNBC cells in vitro and in vivo. Besides, miR-26a-5p negatively regulated the expression of BARD1 and NABP1 and resulted in homologous recombination repair defect (HRD). Notably, overexpressing miR-26a-5p not only facilitated the Olaparib sensitivity of TNBC cells but also the combination of Cisplatin and Olaparib. Furthermore, hormone receptors functioned as transcription factors in the expression of miR-26a-5p, which explained the reasons that miR-26a-5p expressed lowest in TNBC. CONCLUSIONS: Taken together, we reveal the important role of miR-26a-5p in Cisplatin sensitivity and highlight its new mechanism in DNA damage and synthetic lethal.

Craniofacial involvement in Langerhans cell histiocytosis: A review of 44 cases at a single medical center.

BACKGROUND: Craniofacial bones are the most commonly involved site of Langerhans cell histiocytosis (LCH). The main purpose of this study was to clarify the relation between subsites of craniofacial bone and clinical presentation, treatment modalities, outcomes, and permanent consequences (PCs) in patients with LCH. METHODS: Forty-four patients diagnosed with LCH involving the craniofacial region presenting at a single medical center during 2001-2019 were collected and divided into four groups: single system with unifocal bone lesion (SS-LCH, UFB); single system with multifocal bone lesions (SS-LCH, MFB); multisystem without risk organ involvement (MS-LCH, RO-); and multisystem with risk organ involvement (MS-LCH, RO+). Data including demographics, clinical presentation, treatments, outcomes, and the development of PC were retrospectively reviewed. RESULTS: Temporal bone (66.7% versus 7.7%, p = 0.001), occipital bone (44.4% versus 7.7%, p = 0.022), and sphenoid bone (33.3% versus 3.8%, p = 0.041) involvement were more common in SS-LCH, MFB than they were in SS-LCH, UFB. No difference of reactivation rate was noted among the four groups. The most common PC is diabetes insipidus (DI), reported in 9 of the 16 (56.25%) patients with PC. The single system group was reported with the lowest incidence of DI (7.7%, p = 0.035). The reactivation rate was also higher in patients with PC (33.3% versus 4.0%, p = 0.021) or DI (62.5% versus 3.1%, p < 0.001). CONCLUSION: An increased risk of multifocal or multisystem lesions was associated with temporal bone, occipital bone, sphenoid bone, maxillary bone, eye, ear, and oral involvement, which may indicate poor outcomes. Longer follow-up may be indicated if there is the presence of PC or DI due to the high risk of reactivation. Therefore, multidisciplinary evaluation and treatment according to risk stratification are vital for patients diagnosed with LCH involving the craniofacial region.

The Comparison of Functional Outcomes in Patients With Unilateral or Bilateral Intracapsular Mandibular Condylar Fractures After Closed or Open Treatment: A 10-Year Retrospective Study.

BACKGROUND: The treatment strategies for mandibular condylar head fractures, also known as intracapsular condylar fractures (ICFs), have not been concluded. We humbly present our treatment outcomes and share our experience in our department. AIMS AND OBJECTIVES: The aim of this study was to compare the functional outcomes between closed reduction (CR) and open reduction and internal fixation (ORIF) for management of unilateral or bilateral ICFs. MATERIALS AND METHODS: This 10-year retrospective cohort study included 71 patients with 102 ICFs who were treated in our department from May 2007 to August 2017. Nine patients mixed with extracapsular fractures were excluded; thus, 62 patients with 93 ICFs were included. All patients received treatment by the senior surgeon in Chang Gung Memorial Hospital, Linkou Branch, Taiwan. The patient's basic data, fracture morphologies, associated injuries, managements, complications, and maximal mouth opening (MMO) measurement at 1, 3, 6, and 12 months postoperative were reviewed for analysis. RESULTS: Among the 93 fractures, 31 (50%) were bilateral and 31 were unilateral (50%). Based on He's classification, 45 (48%) had type A fracture, 13 (14%) had type B, 5 (5%) had type C, 20 (22%) had type M, and 10 (11%) had no displacement. Maximal mouth opening of 37 mm in unilateral cases after 6 months was significantly higher than the 33-mm MMO in bilateral cases. In addition, the MMO in the ORIF group was significantly higher than that of the CR group in 3 months postoperative. Univariate (odds ratio, 4.92; P = 0.01) and multivariate (odds ratio, 4.76; P = 0.027) analyses revealed CR as an independent risk factor for trismus development compared with ORIF. Malocclusion was observed in 5 patients in both CR and ORIF groups. In addition, 1 patient developed temporomandibular joint osteoarthritis in the CR group. No surgical-related temporary or permanent facial nerve palsy was observed. CONCLUSIONS: Open reduction and internal fixation for condylar head fracture provided better recovery in MMO than CR, and the MMO recovery was less in bilateral condylar head fracture than unilateral condylar head fracture. Open reduction and internal fixation in ICFs have a lower risk for trismus development and should be the treatment of choice in selected cases.

Facial Fracture Patterns and Severity of Associated Injuries After Motorcycle Accidents.

BACKGROUND: Motorcycle injuries remain a significant cause of motor vehicle-related morbidity and mortality in Taiwan. Besides, the facial region is a commonly fractured site under motorcycle accidents. To date, there are no trauma mechanism-based studies of facial fractures. AIM AND OBJECTIVES: This study aims to determine the facial fracture patterns, the severity of head trauma, and associated injuries by different motorcycle-related trauma mechanisms enabling a greater understanding of its distribution and magnitude. METHODS: This is a retrospective descriptive analysis conducted over a 2-year period at Linkou Chang Gung Memorial Hospital. We focused on the population of maxillofacial injury caused by motorcycle accidents. We divided the patient cohort into 3 main trauma mechanisms: single-motorcycle collision (SM group), motorcycle-to-motorcycle collision (MM group), motorcycle-to-vehicle collision (MV group). Data, including demographics, age, fracture patterns of facial bones, and other associated injuries, were collected. RESULTS: A total of 881 cases were identified that involved facial fractures. Most patients were male (71%), young adult (mean age, 32.49 years), and the most common fracture region is the midfacial fracture (79.5%, 700 victims). Among the 3 groups, the MM group was less likely to sustain severe injuries by trauma score system, less head injury and mortality rate. The MV group and SM group have similar mortality rates but different fracture pattern tendencies. Lower facial fractures were more likely in the MV group, but midface fractures in the SM group. Associated injuries were higher in the MV group. CONCLUSIONS: Our study presents the different trends of fracture patterns and injury under 3 main mechanisms of motorcycle casualties. We document all these data in the hope of providing insights into trauma doctors dealing with motorcycle accidents.

Comparison of the Effectiveness Between Conventional and Modified Cinch Suture Techniques in LeFort I Osteotomy: A Systematic Review and Meta-Analysis of Randomized Control Trials.

BACKGROUND: LeFort I osteotomy changes the morphology of the nose. The cinch suture has been proven to prevent the increase in nasal base and alar width. Different types of cinch sutures have been proposed. However, their effectiveness is unclear. AIM AND OBJECTIVES: The aim of this study was to compare the surgical outcomes between conventional and modified cinch techniques through a systematic review and meta-analysis of randomized control trials (RCTs). MATERIAL AND METHODS: We performed systematic search from Embase, PubMed, and the Cochrane Library according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement up to March 2021. The surgical techniques of different cinch sutures were reviewed, and the outcomes of nasal alar width and alar base width were compared between modified and conventional methods. RESULTS: A total of 4 eligible RCTs were included in this meta-analysis. Pooled data showed no significant difference in alar base width change between modified and conventional methods (mean difference, -0.37; 95% confidence interval, -1.32 to 0.57; P = 0.44). Pooled data of 3 studies also showed no significant difference in nasal alar width change (mean difference, -0.11; 95% confidence interval, -1.18 to 0.95; P = 0.83). CONCLUSION: Based on the current data pooled from the available RCTs, no significant difference was found between the conventional cinch technique and the modified technique.

Application of Orthognathic Surgery Principles and Techniques in the Management of Craniomaxillofacial Trauma.

BACKGROUND: Posttraumatic secondary deformities of facial skeleton may occur because of nonmanagement or ill management of primary injuries, whereas some unexpectedly occur even after dedicated management attempts. Orthognathic surgery (OGS) principles and techniques can be used as an efficient tool to correct posttraumatic craniomaxillofacial deformities or skeletal developmental deformities during trauma management. AIM AND OBJECTIVES: The aims of this study were to describe the different types of posttraumatic secondary deformity and address how to use the principles of orthognathic techniques to correct them. MATERIALS AND METHODS: Patients with orthognathic surgical osteotomies during primary or secondary management of facial trauma (referred to as trauma-OGS) during the period of 2010 to 2018 were retrospectively reviewed. Variables pertaining to patients and surgery were collected, including trauma diagnosis, etiology, duration between trauma/primary surgery and secondary presentation, suggested reason for secondary deformity, intervention undertaken to address, and the surgical outcome. RESULTS: Twenty-seven patients were eligible and extensively reviewed. Etiological categorization of trauma-OGS could be done into posttraumatic deformities (18) and developmental deformities (9). The former group was further categorized as OGS done as primary procedure (8) that included immobile Le Fort fractures and delayed initial treatment, and OGS done as secondary procedure (10) that included complex fractures and condylar fractures. The developmental deformity group was categorized into OGS done simultaneously during trauma management (5) or done as a secondary procedure after trauma management (4). CONCLUSIONS: Application of principles and techniques of OGS in indicated primary or secondary management of facial trauma patients should always be considered. The categorization of scenarios presented in this article relating facial trauma and OGS may further help to understand the application.

Aesthetic Outcome of Primary Rhinoplasty of Saddle Nose Deformity in Naso-Orbital Ethmoidal Fractures in Asian Patients.

BACKGROUND: Saddle nose deformity following naso-orbital ethmoidal (NOE) fractures remain a challenging problem for the reconstructive surgeon. Early reduction and internal fixation allow for fracture stabilization but is unable to address the problem of the depressed nasal dorsum, especially after soft tissue shrinkage. The aim of this study is to evaluate the outcome of primary rhinoplasty in patients with NOE fractures. MATERIALS AND METHODS: From 2016 to 2019, 9 patients presented to our department with NOE fractures complicated by saddle nose deformity underwent primary nasal reconstruction at the time of their fracture fixation. Life size (1:1) frontal and lateral postoperative photographs were taken. Three objective measurements were made, including the nasofrontal angle, tip projection, and radix projection. These measurements were compared between normal persons (group 1), preoperative patients (group 2), and postoperative patients (group 3). Nose aesthetic assessment was carried out via a panel assessment using a Visual Analog Scale of 5. Patient satisfaction was further assessed subjectively by the patient themselves using the Visual Analog Scale. RESULTS: When comparing group 3 to 2, a significant reduction in the nasofrontal angles was found with an accompanying increase in the radix and tip projection ( P <0.05). No statistical significance between normal persons and postprimary rhinoplasty patients was noted between groups 1 and 3. Average patient satisfaction scored 3.86±1.07 compared with 3.63±0.84 by laypersons and 4±0.77 by specialists' panel. CONCLUSION: Primary nasal reconstruction may be an alternative method for achieving optimum results following NOE fractures preventing the development of secondary saddle nose deformity with a shortened nose which may potentially be more difficult to correct.

Human NOP2/NSUN1 regulates ribosome biogenesis through non-catalytic complex formation with box C/D snoRNPs.

5-Methylcytosine (m5C) is a base modification broadly found on various RNAs in the human transcriptome. In eukaryotes, m5C is catalyzed by enzymes of the NSUN family composed of seven human members (NSUN1-7). NOP2/NSUN1 has been primarily characterized in budding yeast as an essential ribosome biogenesis factor required for the deposition of m5C on the 25S ribosomal RNA (rRNA). Although human NOP2/NSUN1 has been known to be an oncogene overexpressed in several types of cancer, its functions and substrates remain poorly characterized. Here, we used a miCLIP-seq approach to identify human NOP2/NSUN1 RNA substrates. Our analysis revealed that NOP2/NSUN1 catalyzes the deposition of m5C at position 4447 on the 28S rRNA. We also find that NOP2/NSUN1 binds to the 5'ETS region of the pre-rRNA transcript and regulates pre-rRNA processing through non-catalytic complex formation with box C/D snoRNAs. We provide evidence that NOP2/NSUN1 facilitates the recruitment of U3 and U8 snoRNAs to pre-90S ribosomal particles and their stable assembly into snoRNP complexes. Remarkably, expression of both WT and catalytically inactive NOP2/NSUN1 in knockdown background rescues the rRNA processing defects and the stable assembly of box C/D snoRNP complexes, suggesting that NOP2/NSUN1-mediated deposition of m5C on rRNA is not required for ribosome synthesis.

Preliminary outcomes of the surgical navigation system combined with intraoperative three-dimensional C-arm computed tomography for zygomatico-orbital fracture reconstruction.

This study analyzed the outcomes of zygomatico-orbital fracture reconstruction using the real-time navigation system with intraoperative three-dimensional (3D) C-arm computed tomography (CT). Fifteen patients with zygomatico-orbital or isolated orbital/zygoma fractures were enrolled in this prospective cohort. For zygoma reduction, the displacement at five key sutures and the differences between preoperative and intraoperative CT images were compared. For orbital reconstruction, the bilateral orbital volume differences in the anterior, middle, and posterior angles over the medial transitional buttress were measured. Two patients required implant adjustment once after the intraoperative 3D C-arm assessment. On comparing the preoperative and postoperative findings for the zygoma, the average sum of displacement was 19.48 (range, 5.1-34.65) vs. 1.96 (0-3.95) mm (P < 0.001) and the deviation index was 13.56 (10-24.35) vs. 2.44 (0.6-4.85) (P < 0.001). For the orbit, the mean preoperative to postoperative bilateral orbital volume difference was 3.93 (0.35-10.95) vs. 1.05 (0.12-3.61) mm3 (P < 0.001). The mean difference in the bilateral angles at the transition buttress was significantly decreased postoperatively at the middle and posterior one-third. There was no significant difference in orbital volume, angle of the transition zone, and the sum of five zygoma distances between post operative results and preoperative virtual planning. The surgical navigation system with the intraoperative 3D C-arm can effectively improve the accuracy of zygomatico-orbital fracture reconstruction and decrease implant adjustment times.

Outcome Analysis of Surgical Timing in Pediatric Orbital Trapdoor Fracture with Different Entrapment Contents: A Retrospective Study.

Orbital trapdoor fracture occurs more commonly in pediatric patients, and previous studies suggested early intervention for a better outcome. However, there is no consensus on the appropriate timing of emergent intervention due to the insufficient cases reported. In the current retrospective study, we compared the outcomes of patient groups with different time intervals from injury to surgical intervention and entrapment content. Twenty-three patients who underwent surgery for trapdoor fracture between January 2001 and September 2018 at Chang Gung Memorial Hospital were enrolled. There was no significant difference in diplopia and extraocular muscle (EOM) movement recovery rate in patients who underwent surgery within three days and those over three days. However, among the patients with an interval to surgery of over three days, those with muscle entrapment required a longer period of time to recover from EOM movement restriction (p = 0.03) and diplopia (p = 0.03) than those with soft tissue entrapment. Regardless of time interval to surgery, patients with muscle entrapment took longer time to recover from EOM movement restriction (p = 0.036) and diplopia (p = 0.042) and had the trend of a worse EOM recovery rate compared to patients with soft tissue entrapment. Hence, we suggested that orbital trapdoor fractures with rectus muscle entrapment should be promptly managed for faster recovery.

Target-Based Virtual Screening and LC/MS-Guided Isolation Procedure for Identifying Phloroglucinol-Terpenoid Inhibitors of SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to more than 5 million deaths worldwide to date. Due to the limited therapeutic options so far available, target-based virtual screening with LC/MS support was applied to identify the novel and high-content compounds 1-4 with inhibitory effects on SARS-CoV-2 in Vero E6 cells from the plant Dryopteris wallichiana. These compounds were also evaluated against SARS-CoV-2 in Calu-3 cells and showed unambiguous inhibitory activity. The inhibition assay of targets showed that compounds 3 and 4 mainly inhibited SARS-CoV-2 3CLpro, with effective Kd values. Through docking and molecular dynamics modeling, the binding site is described, providing a comprehensive understanding of 3CLpro and interactions for 3, including hydrogen bonds, hydrophobic bonds, and the spatial occupation of the B ring. Compounds 3 and 4 represent new, potential lead compounds for the development of anti-SARS-CoV-2 drugs. This study has led to the development of a target-based virtual screening method for exploring the potency of natural products and for identifying natural bioactive compounds for possible COVID-19 treatment.

Application of real-time surgical navigation for zygomatic fracture reduction and fixation.

BACKGROUND: Inappropriate treatment of zygomatic fractures can reduce esthetic and functional outcomes. The aim of this study was to answer the research question: "Among patients with a unilateral zygomatic fracture, is the use of computer-assisted real-time navigation system during fracture reduction precise and accurate to create postoperative facial symmetry?" METHODS: Using a retrospective cohort study design, we enrolled a cohort of unilateral zygomatic fractures undergoing open reduction and internal fixation (ORIF) with the aid of the computer-based navigation system at Chang Gung Memorial Hospital, Taiwan, during January 2015 and March 2018. The predictor variable was the comparison before and after surgery. The main outcome variables included (1) two-dimensional (2D) reduction of the displacement at five anatomical landmarks: zygomaticofrontal, inferior orbital rim, zygomaticosphenoidal, zygomaticomaxillary, and zygomaticotemporal lines/buttresses and (2) three-dimensional (3D) differences on distances between zygomatic surface to the porion plane and the midpoint of zygomatic arch (ZA) to the mid-porion (MP) plane. The Wilcoxon signed-rank test was computed to compare between pre- and postoperative data, and a p-value less than 0.05 was considered statistically significant. RESULTS: The cohort comprised 24 subjects (50% females, 75% left-sided) with a mean age of 30.5 +/- 13.8 years. On 2D analysis, the significant fracture reduction was found: 4.78 vs. 1.22 mm, 1.78 vs. 0.40 mm, 3.50 vs. 0.07 mm, 3.06 vs. 0.55 mm, and 2.55 vs. 0.50 mm at zygomaticomaxillary, zygomaticofrontal, inferior orbital rim, zygomaticosphenoidal, and zygomaticotemporal landmarks. The 3D evaluations revealed the significant reduction of the differences between the left and right zygomatic surface to the porion plane (4.09 ± 2.12 vs. 0.46 ± 0.35 mm) and between the left and right ZA midpoints to the MP plane (4.89 ± 2.59 vs. 0.71 ± 0.44 mm) (p<0.001 for both 2D and 3D analyses). CONCLUSIONS: The results of this study suggest that the real-time surgical navigation system can effectively guide the ORIF of zygomatic fractures. Future research studies should focus on the learning curve and cost-effectiveness analysis of this technique.

Lower Eyelid and Midface Rejuvenation: Suborbicularis Oculi Fat Lift.

The lower eyelids and midface are considered to be a contiguous aesthetic unit although they are different anatomic structures. Through in-depth understanding of complex anatomy and aging theory and appropriate surgical strategies according to the type of aging, surgical outcome of aging lower eyelid/midface can be more and more predictable. This article discusses the characteristics and theories of aging and 5 types of lower eyelid/middle face aging based on 4 key factors, namely, protruding fat in the orbital, excess skin on the lower eyelid, sagging midface and soft tissue deflation. Various combinations of surgical strategies are adopted accordingly.

p53 mitigates the effects of oncogenic HRAS in urothelial cells via the repression of MCOLN1.

Inhibition of TRPML1, which is encoded by MCOLN1, is known to deter cell proliferation in various malignancies. Here, we report that the tumor suppressor, p53, represses MCOLN1 in the urothelium such that either the constitutive loss or ectopic knockdown of TP53-in both healthy and bladder cancer cells-increased MCOLN1 expression. Conversely, nutlin-mediated activation of p53 led to the repression of MCOLN1. Elevated MCOLN1 expression in p53-deficient cancer cells, though not sufficient for bolstering proliferation, augmented the effects of oncogenic HRAS on proliferation, cytokine production, and invasion. Our data suggest that owing to derepression of MCOLN1, urothelial cells lacking p53 are poised for tumorigenesis driven by oncogenic HRAS. Given our prior findings that HRAS mutations predict addiction to TRPML1, this study points to the utility of TRPML1 inhibitors for mitigating the growth of a subset of urothelial tumors that lack p53.

p53 induces a survival transcriptional response after nucleolar stress.

Accumulating evidence indicates that increased ribosome biogenesis is a hallmark of cancer. It is well established that inhibition of any steps of ribosome biogenesis induces nucleolar stress characterized by p53 activation and subsequent cell cycle arrest and/or cell death. However, cells derived from solid tumors have demonstrated different degrees of sensitivity to ribosome biogenesis inhibition, where cytostatic effects rather than apoptosis are observed. The reason for this is not clear, and the p53-specific transcriptional program induced after nucleolar stress has not been previously investigated. Here we demonstrate that blocking rRNA synthesis by depletion of essential rRNA processing factors such as LAS1L, PELP1, and NOP2 or by inhibition of RNA Pol I with the specific small molecule inhibitor CX-5461, mainly induce cell cycle arrest accompanied by autophagy in solid tumor-derived cell lines. Using gene expression analysis, we find that p53 orchestrates a transcriptional program involved in promoting metabolic remodeling and autophagy to help cells survive under nucleolar stress. Importantly, our study demonstrates that blocking autophagy significantly sensitizes cancer cells to RNA Pol I inhibition by CX-5461, suggesting that interfering with autophagy should be considered a strategy to heighten the responsiveness of ribosome biogenesis-targeted therapies in p53-positive tumors.

Le Fort Fractures With Maxillary Immobility: Classification and the Moment Concept to Rationalize Optimal Surgical Treatment.

BACKGROUND: Le Fort fractures with maxillary immobility are an uncommon presentation of facial trauma often associated with the disturbance of occlusion. Historically, cases involving high Le Fort fractures require a coronal extensive approach to mobilize the maxilla for occlusion restoration. Here, we review our institutional series of immobile Le Fort fractures and present their treatment approach, outcomes, classification, and then rationalize our treatment with a concept philosophy. MATERIALS AND METHODS: We treated 12 consecutive patients with unilateral and bilateral immobile Le Fort fractures from 2010 to 2017. The mean age was 27.1 years. Ten patients had a unilateral greenstick Le Fort III fracture. Five patients also had associated mandible fractures. Intraoperative occlusions could only be restored after Le Fort I osteotomy was performed on the side of the immobile Le Fort fracture. The mean follow-up period was 1.6 years. RESULTS: All patients presented good long-term occlusion restoration. There were no cases of nonunion or significant complications. None of the patients complained of any malar or periorbital cosmetic issues during follow-up sessions. CONCLUSIONS: We presented an institutional series and classification of unusual Le Fort fractures with maxillary immobility, including a previously unreported fracture configuration of greenstick unilateral Le Fort III fractures combined with complete contralateral Le Fort I fracture. Based on our moment concept to rationalize treatment, as well as a systematic review of published literature, we advocate the judicious use of Le Fort I osteotomy to efficiently and safely treat these unusual fractures.

Overexpression of miRNA-3613-3p Enhances the Sensitivity of Triple Negative Breast Cancer to CDK4/6 Inhibitor Palbociclib.

Triple negative breast cancer (TNBC) is characterized by lack of expression of the estrogen and progesterone receptors and HER2, which are common therapeutic targets. CDK4/6 inhibitor Palbociclib has been approved as an anti-cancer agent for breast cancer. However, identifying biomarkers that predict the response to Palbociclib has always been a challenge for molecular targeted therapy. In this study, we identify microRNA as a hallmark in TNBC patients and explore if miR-3613-3p might serve as a tumor suppressor biomarker for triple negative breast cancer patients and if overexpression of miR-3613-3p could enhance the sensitivity of TNBC cells to Palbociclib. We show that the expression of miR3613-3p was down-regulated in TNBC tumors and cells, and the overexpression of miR-3613-3p in patients' tumor tissues was clinically and pathologically correlated with favorable prognosis, such as smaller tumor size and the lower Ki-67. In vitro, overexpression of miR-3613-3p inhibited cell proliferation, induced G1 cell-cycle arrest, and enhanced the sensitivity of TNBC cells to Palbociclib treatment. In vivo study revealed that overexpression of miR-3613-3p inhibited TNBC tumorigenesis and exerted a significant inhibitory effect of Palbociclib on MDA-MB-231 cells. Mechanically, SMAD2 and EZH2 were found to be two direct targets of miR-3613-3p and mediate the proliferation of TNBC cells and the sensitivity of the cells to Palbociclib through inducing cellular senescence. Our findings suggested that miR-3613-3p acts as a cancer-suppressor miRNA in TNBC. Moreover, our study showed that miR-3613-3p might be used as a predictive biomarker for the response of TNBC to Palbociclib.