Metaproteomics Characterizes the Human Gingival Crevicular Fluid Microbiome Function in Periodontitis.

Periodontitis is the leading cause of tooth loss in adults worldwide. The human proteome and metaproteome characterization of periodontitis is not clearly understood. Gingival crevicular fluid samples were collected from eight periodontitis and eight healthy subjects. Both the human and microbial proteins were characterized by liquid chromatography coupled with high-resolution mass spectrometry. A total of 570 human proteins were found differentially expressed, which were primarily associated with inflammatory response, cell death, cellular junction, and fatty acid metabolism. For the metaproteome, 51 genera were identified, and 10 genera were found highly expressed in periodontitis, while 11 genera were downregulated. The analysis showed that microbial proteins related to butyrate metabolism were upregulated in periodontitis cases. In particular, correlation analysis showed that the expression of host proteins related to inflammatory response, cell death, cellular junction, and lipid metabolism correlates with the alteration of metaproteins, which reflect the changes of molecular function during the occurrence of periodontitis. This study showed that the gingival crevicular fluid human proteome and metaproteome could reflect the characteristics of periodontitis. This might benefit the understanding of the periodontitis mechanism.

Proteome Characterization of Glaucoma Aqueous Humor.

Glaucoma is the leading cause of irreversible blindness worldwide. The proteome characterization of glaucoma is not clearly understood. A total of 175 subjects, including 57 primary acute angle-closure glaucoma (PAACG), 50 primary chronic angle-closure glaucoma (PCACG), 35 neovascular glaucoma (NVG), and 33 cataract patients, were enrolled and comparison proteomic analysis was provided. The samples were randomly divided into discovery group or validation group, whose aqueous humor proteome was analyzed by data-independent acquisition or by parallel reaction monitoring. The common proteome features of three types of glaucoma were immune response, lipid metabolism, and cell death. Three proteins, VTN, SERPIND1, and CD14, showed significant upregulation in glaucoma and could discriminate glaucoma from cataract. Mutual differential proteomic analysis of PAACG, PCACG, and NVG showed different proteome characterization of the three types of glaucoma. NVG was characterized with activated angiogenesis. PAACG was characterized with activation of inflammation response. SERPIND1 was discovered to play vital role in glaucoma occurrences, which is associated with eye transparency decrease and glucose metabolism. This study would provide insights in understanding proteome characterization of glaucoma and benefit the clinical application of AH proteome.

A qualitative and quantitative analysis of the human gingival crevicular fluid proteome and metaproteome.

Gingival crevicular fluid (GCF) is an integral part of oral fluid that plays a special role in maintaining the structure of junctional epithelium and defending against bacterial infection. In this study, we comprehensively analysed the composition of the human GCF proteome and metaproteome simultaneously to obtain multidimensional information about GCF. A total of 3680 human proteins (2540 with at least two unique peptides) were identified in the normal GCF sample, and their functions were mainly associated with immune function and inflammation. Among these proteins, 1874 proteins could be quantified by the iBAQ algorithm, and their abundances spanned a dynamic range of six orders of magnitude. For the GCF metaproteome, a total of 3082 proteins and 69 genera were found. In addition, 16 genera were not identified by GCF metagenomic analysis. Compared to the saliva metaproteome, 32 genera were found to be in common. The protein quantitative analysis showed that the abundance of GCF metaproteome contributed to approximately 4.17% of the total GCF proteome. The top three most abundant genera were Fusobacterium, Corynebacterium, and Leptotrichia. The above data will be useful for future research on GCF-related diseases.

Self-synthesized extracellular matrix contributes to mature adipose tissue regeneration in a tissue engineering chamber.

The development of an engineered adipose tissue substitute capable of supporting reliable, predictable, and complete fat tissue regeneration would be of value in plastic and reconstructive surgery. For adipogenesis, a tissue engineering chamber provides an optimized microenvironment that is both efficacious and reproducible; however, for reasons that remain unclear, tissues regenerated in a tissue engineering chamber consist mostly of connective rather than adipose tissue. Here, we describe a chamber-based system for improving the yield of mature adipose tissue and discuss the potential mechanism of adipogenesis in tissue-chamber models. Adipose tissue flaps with independent vascular pedicles placed in chambers were implanted into rabbits. Adipose volume increased significantly during the observation period (week 1, 2, 3, 4, 16). Histomorphometry revealed mature adipose tissue with signs of adipose tissue remolding. The induced engineered constructs showed high-level expression of adipogenic (peroxisome proliferator-activated receptor γ), chemotactic (stromal cell-derived factor 1a), and inflammatory (interleukin 1 and 6) genes. In our system, the extracellular matrix may have served as a scaffold for cell migration and proliferation, allowing mature adipose tissue to be obtained in a chamber microenvironment without the need for an exogenous scaffold. Our results provide new insights into key elements involved in the early development of adipose tissue regeneration.

Impact of Two Urethral Stent Types on the Complications After One-Stage Hypospadias Repair Using the Duckett Procedure: A retrospective cohort study.

PURPOSE: The purpose of this study was to evaluate whether the new nickel-titanium alloy stents are superior to traditional silicone stents in hypospadias repair surgery to prevent complications such as urinary fistula. METHODS: This retrospective cohort study included 576 patients with hypospadias who underwent the placement either with nickel-titanium alloy stents or traditional silicone stents after hypospadias surgery between March 2002 and August 2019. The patients were assigned into the nickel-titanium alloy stent group (group NTAS) and the silicone stent group (group SS). The primary outcome was assessed with the rate of urinary fistula occurrence at four weeks (stent removal time), and the secondary outcomes were decided on the rate of other complications such as urethral stricture, and urethral diverticulum, infection, etc. The occurrence of complications in both groups was compared and the important contributing factors of urinary fistula and urethral stricture were determined. RESULTS: Among 576 patients, 398 were assigned into group NTAS while 178 were into group SS. 35 patients in the group NTAS and 30 in the group SS developed urinary fistula with a ratio of 8.8% and 16.9%, respectively (p = 0.005). Subgroup analysis showed that the differences were mainly in preschool patients (≤ 6 years) (p = 0.004) and those with the penile type of hypospadias (p = 0.008). In addition, urethral stricture complicated five patients in the NTAS group and two in the SS group with a ratio of 1.3% and 1.1%, respectively (p = 1.000). Logistic regression showed that hypospadias type (p = 0.001) and stent type (p = 0.001) are the important risk factors for urethral fistula. CONCLUSIONS: Nickel-titanium alloy stents reduced the occurrence of urinary fistula complications after hypospadias repair in preschool patients, and can be optioned as a better choice for hypospadias surgery.

Physiochemical responses of C. elegans under exposure to lanthanum and cerium affected by bacterial metabolism.

The increasing demand for rare earth elements (REEs) in modern applications has drawn significant attention. REEs can be introduced into the environment through REE-containing fertilizers, abandoned REE-rich equipment, and mining, persisting and impacting soil quality, nutrient cycles, and plant growth. Scientists have raised concerns about REEs entering the food chain from the environment and eventually accumulating in organisms. Decades of experimental evidence have shown that these effects include inhibited growth, impaired liver function, and alterations in children's intelligence quotients. However, there exists a paucity of research that has elucidated the metabolic-level biological impacts of REEs. In our study, Caenorhabditis elegans (C. elegans) was used as a model organism to investigate physiological and inherent metabolic changes under exposure to different concentrations of REEs. The diet bacteria of nematodes play a key role in their life and development. Therefore, we investigated the influence of bacterial activity on the nematodes' response to REE exposure. We observed a concentration-dependent accumulation of REEs in nematodes, which consequently led to a reduction in lifespan and alterations in body length. Exposure to a mixed solution of REEs, in comparison to a single REE solution, resulted in greater toxicity toward nematodes. The metabolic results showed that the above changes were closely related to REE-induced amino acid metabolism disorder, membrane disturbance, DNA damage, and oxidative stress. Of note, the presence of living bacteria elicits REE effects in C. elegans. These findings highlight the potential intrinsic metabolic changes occurring in nematodes under REE exposure. Our study raises awareness of the exposure risks associated with REEs, provides valuable insight into the metabolic-level biological impacts of REEs and contributes to the development of effective mitigation strategies to reduce potential risks to human health.

96DRA-Urine: A high throughput sample preparation method for urinary proteome analysis.

Mass spectrometry (MS)-based urinary proteomics is increasingly used for clinical research. A critical step in urinary proteomic analysis comprises the implementation of a reliable sample preparation method with high yields of peptides and proteins. In this study, we developed a urinary sample preparation method, DRA-Urine (Direct reduction/alkylation in urine), which urinary proteins were directly reduced/alkylated in urine, and then precipitated by acetone, washed and digestion on an ultrafilter unit. The qualitative and quantitative comparison of different urinary sample preparation methods (in-solution methods and ultrafilter-assisted methods) showed that DRA-Urine could achieve better results. Adapting DRA-Urine protocol to a 96-well format, namely 96DRA-Urine, shortened the time for buffer change and improved sample preparation throughput. The results showed that 96DRA-Urine displayed similar proteomic performance to DRA-Urine. Finally, the 96DRA-Urine method was used in a label-free, small pilot biomarker discovery analysis for differential urinary proteome analysisof bladder cancer urine. The results showed that urinary proteins could differentiate bladder cancer (BCa) patients from healthy controls and distinguish high-grade BCa from low-grade BCa with area under the curve (AUC) values of 0.972 and 0.847, respectively. Consequently, the 96DRA-Urine method might be a high-throughput method for preparing body fluid samples used in clinical research but needs to be further verified.

Experience with flap repair after vulvar carcinoma resection: a retrospective observational study of 26 cases.

Background: A defect caused by the radical resection of vulvar cancer requires repair with flap transplantation or vulvoplasty. However, in clinical practice, the surgeons encounter difficulties while using a flap to repair the wound. Therefore, this study aimed to present a review of our practice of post-surgical defect reconstruction in cases using different skin flaps. Methods: An observational study was performed involving 26 patients with vulvar cancer who were admitted to Sun Yat-Sen Memorial Hospital between February 2015 and February 2020 for surgical and reconstructive procedures. The clinical data of these 26 patients were analyzed. All patients underwent radical resection of vulvar cancer, followed by post-surgical defect repair using random flap or axial flap transplantation (even for very complex defects). The clinical variables collected and the assessment of efficacy included survival of the flap, history of dysfunction of the recipient area, such as scar contracture, and satisfaction of the patient with the shape after external vaginal surgery. Results: Among the 26 cases in this study, all patients underwent 38 soft tissue reconstruction procedures for vulvar perineal defects during the study period. Squamous cell carcinoma was the most commonly diagnosed cancer (80.8%). The average size of the defect was 9.3×7 cm2. Rhomboid flaps were the most commonly used flaps for performing reconstruction in both the primary and recurrent groups. Poor wound healing was the most commonly discovered complication, which occurred in three of the 38 flaps (7.9%) used. Previous surgery or radiotherapy did not increase the rate of complications following successful reconstruction. Conclusions: Different skin flaps are effective premium options for post-surgical defect reconstruction, and the selective use of skin flaps for treating vulvar defects preserves the vulvar morphology and allows for relatively better functionality.

Clinical use of magnetic resonance imaging in buttock augmentation with silicone implants: a retrospective analysis.

Background: The objective of the current study was to retrospectively examine the morphological magnetic resonance imaging (MRI) characteristics of the gluteus maximus of buttock augmentation at levels of predetermined anatomic points. Methods: The present study was a retrospective cross-sectional study. Adult women who underwent high-quality MRI scanning at Sun Yat-sen Memorial Hospital of Sun Yat-sen University from January 2018 to January 2021 were included in this work. The transverse MRI data measured at the inferior point of the sacroiliac joint, just above the femoral head, and at the ischial tuberosity were collected and statistically analyzed. Results: Fifty-two cases (104 sides of female gluteus maximus) were included in the final analysis. The A point (surgery starting point) were 54.4±6.34 mm, 54.91±5.57 mm, and 73.91±5.57 mm away from the posterior midline at the level of inferior point of the sacroiliac joint, just above the femoral head, and at the ischial tuberosity, respectively. Accordingly, the thickness of the muscle at these locations was 16.0±4.17 mm, 23.4±4.40 mm, and 24.6±7.58 mm, respectively. The diameter of the implant did not exceed 14.18±1.22 cm. In addition, the gluteus maximus at the lowest point of the sacroiliac joint and above the femoral head exhibited an arc structure, which needs to be tilted to the deep plane during separation. Conclusions: Dissimilar from previous experience of blind dissection, the gluteus maximus muscle can be more scientifically and reasonably dissected using the indexes for gluteus augmentation supplied in this study.

Qualitative and quantitative proteomic and metaproteomic analyses of healthy human urine sediment.

PURPOSE: The healthy human urine sediment proteome and metaproteome are investigated, to shed light on the variations of urine sediment proteins and metaproteins associated with sex and age. EXPERIMENTAL DESIGN: Urine sediment samples are collected from 19 healthy subjects. Protein identification and quantification are performed by liquid chromatography coupled high-resolution mass spectrometry. RESULTS: A total of 2736 human proteins were identified, which were primarily associated with inflammatory response and energy metabolism. For the metaproteome, 65 genera were identified that were primarily involved in translation and carbohydrate metabolic processes. The median biological coefficient variation of the proteome/metaproteome of human urine sediment was 0.5/0.72, similar to the proteome of human urine supernatant. In addition, sex and age were observed to affect the proteome and metaproteome of healthy human urine sediment. CONCLUSION AND CLINICAL RELEVANCE: The healthy human urine sediment were characterized, indicating that urine sediment might represent an alternative resource for disease research in addition to urine supernatant, but the influence of sex and age must be considered in the study design process.

Ear Reconstruction with the Combination of Expanded Skin Flap and Medpor Framework: 20 Years of Experience in a Single Center.

BACKGROUND: In ear reconstruction, the difficulty lies in reestablishing the ear's bionic form with adequate skin coverage and an appropriate framework. Skin expansion and a porous polyethylene (i.e., Medpor) framework are often used for ear reconstruction. However, a long-term review of the combined application of the expanded skin and Medpor framework has not been reported. This article reviews ear reconstruction combining these two factors over the past 20 years in the authors' center to summarize the surgical technique and analyze the postoperative results and complications. METHODS: A retrospective review was performed that included all patients who underwent ear reconstruction with expanded skin and Medpor framework in the authors' center between 1998 and 2018. RESULTS: A total of 68 patients with microtia who were admitted to the authors' center for surgical ear reconstruction were included, and 70 ears were reconstructed. Fifty-seven of the patients (83.82 percent) felt satisfied with their reconstructed ear, five patients (7.35 percent) were not satisfied with the reconstructed ear, and six patients (8.82 percent) had the frameworks removed. Fifteen patients (22.06 percent) developed complications, including framework exposure (13.24 percent), infection (4.41 percent), scar hypertrophy (4.41 percent), and hematoma (2.94 percent). CONCLUSIONS: Framework exposure limits the combined application of expanded skin flap and Medpor framework when reconstructing the ear without additional fascial interposition. Using a temporoparietal fascia or postauricular fascia flap during the operation is effective to decrease the exposure rate; however, this complication cannot be completely avoided. Using postauricular fascia and skin graft may lead to scar hypertrophy; thus, these techniques should be used with caution. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Experiences with Surgical Reconstruction of Penoscrotal Extramammary Paget's Disease: A review of 21 cases.

BACKGROUND: Penoscrotal extramammary Paget's disease is a rarely found intraepithelial adenocarcinoma in older male patients. The challenge for surgeons to perform high-quality post-surgery reconstruction is mostly case-dependent. Here, we present a review of post-surgical reconstruction practice in 21 cases using various scrotal skin flaps. METHODS: During the 10-year study period (from 2008 to 2018), all 21 male patients diagnosed with penoscrotal extramammary Paget's diseases at Sun Yat-sen memorial hospital were enrolled in this study. All patients underwent expanding resections guided by frozen sections and penoscrotal reconstructions using various scrotal skin flaps alone or combined with pubic skin flaps and partial thickness skin graft depending on individual defects. The patient's clinical data, specifically the diagnosis, surgical outcome, and follow-up evaluation were retrospectively analyzed. RESULTS: The size of skin lesions ranged from 4 × 5 cm to 16 × 18 cm approximately. Out of 21 total cases, 13 patients received combined scrotal and pubic flaps, three patients received single scrotal flaps, and five patients received scrotal flaps and partial thickness skin grafts. Limited flap tip necrosis (two cases) and wound dehiscence (one case) were observed, and secondary resections were performed in four cases with positive post-operative pathological results. All patients experienced complete wound healing and had well-maintained penile morphology and function. CONCLUSION: Various scrotal skin flaps alone or combined with pubic skin flaps designed for individual lesion conditions in patients with penoscrotal extramammary Paget's disease are efficient for scrotal reconstruction. A good match of the local skin texture helps to maintain the morphology and function of the penis and scrotum.

Characterization of long non-coding RNA and messenger RNA profiles in laryngeal cancer by weighted gene co-expression network analysis.

Laryngeal cancer (LC) is a malignant tumor in the head and neck region. It was recently elucidated that long non-coding RNAs (lncRNAs) participate in the pathogenesis of LC. However, the detailed mechanism of lncRNA in LC and whether long non-coding RNAs serve as effective biomarkers remains unclear. Ribonucleic acid (RNA) sequence data of LC and 11 patient clinical traits were extracted from The Cancer Genome Atlas (TCGA) database and analyzed by weighted gene co-expression network analysis (WGCNA). A total of 9 co-expression modules were identified. The co-expression Pink module significantly correlated with four clinical traits, including history of smoking, lymph node count, tumor status, and the success of follow-up treatment. Based on the co-expression Pink module, lncRNA-microRNA (miRNA)-messenger RNA (mRNA) and lncRNA-RNA binding protein-mRNA networks were constructed. We found that 8 lncRNAs significantly impacted overall survival (OS) in LC patients. These identified lncRNA and hub gene biomarkers were also validated in multiple LC cells in vitro via qPCR. Taken together, this study provided the framework of co-expression gene modules of LC and identified some important biomarkers in LC development and disease progression.

Transferring the exudate in the tissue engineering chamber as a trigger to incubate large amount adipose tissue in remote area.

Tissue engineering chamber (TEC) is a technique that could incubate up to 16 folds volume increase of a fat flap. But the mechanism in the silicone chamber was still unknown. The function of exudate in the chamber was noticed recently. We developed a special model called fluid drainage model (FDM) that consisted of a traditional TEC on the back and paired fat flaps without a chamber in the groins. Then we used a silicon tube to dynamically transfer the exudate from dorsal TEC to one of the paired inguinal fat flap and the other inguinal fat flap with a sham tube was set as control. At Week 4, the volume of drainage group reached 8.7 ± 2.3 ml, 576 ± 152% to its original volume whereas the growth ratio of control group was only 130 ± 39%. Similar volume change and histological change were observed within fat flap from TEC model and drainage group. The exudate in the TEC is a heterogeneous cocktail contains cytokines as well as cells. Intriguingly, transferred exudate in the TEC model sustain the ability to incubate large amount of adipose tissue remotely.

Adipose stem cell-laden injectable thermosensitive hydrogel reconstructing depressed defects in rats: filler and scaffold.

Facial depressed defects are a common cosmetic problem. Temporary fillers need to be re-injected frequently to maintain the desired outcomes. Here, the feasibility of a novel type of injectable hydrogel for persistent effect is demonstrated. We first useed agmatine to synthesize a poly(amidoamine) (PAA) to form a cell-attachable crosslinker and then the crosslinker was co-polymerized with N-isopropylacrylamide to obtain an injectable and temperature sensitive hydrogel. 1H NMR showed the successful synthesis of the crosslinker. In vitro tests, CCK-8 assay and live/dead viability test showed that the hydrogel was non-toxic to adipose-derived stem cells (ASCs). SEM images also confirmed that ASCs could adhere to the hydrogel. Then we constructed a novel depressed defect model in rats and injected four different fillers in the depressed defects: (1) the hydrogel with ASCs, (2) the hydrogel only, (3) hyaluronic acid, and (4) PBS. After 4 weeks, gross and histological analyses showed the defects in hydrogel, hydrogel + ASCs, and HA groups improved significantly and there were no significant differences among them. Significant differences in thickness from skin to muscle in the defect was found between the hydrogel + ASCs group and the other groups after 6 months. The hydrogels degraded completely in defects in both the hydrogel group and the hydrogel + ASCs group, and were filled with adipocytes and multilocular immature adipocytes. Immunohistochemical study using s-100 and perilipin staining revealed adipocyte differentiation in the defect sites. We also used green fluorescent protein (GFP)-ASCs for tracing and found that exogenous added ASCs were involved in adipogenesis. In conclusion, such a cell attachable thermosensitive hydrogel has definite potential not only as a filler but also as a scaffold, and has a persistent effect for small depressed defects. It might ultimately become a new material in plastic and reconstructive surgery.