Amide Proton Transfer-Weighted Magnetic Resonance Imaging for Application in Renal Fibrosis: A Radiological-Pathological-Based Analysis.

INTRODUCTION: Renal fibrosis (RF), being the most important pathological change in the progression of CKD, is currently assessed by the evaluation of a biopsy. This present study aimed to apply a novel functional MRI (fMRI) protocol named amide proton transfer (APT) weighting to evaluate RF noninvasively. METHODS: Male Sprague-Dawley (SD) rats were initially subjected to bilateral kidney ischemia/reperfusion injury (IRI), unilateral ureteral obstruction, and sham operation, respectively. All rats underwent APT mapping on the 7th and 14th days after operation. Besides, 26 patients underwent renal biopsy at the Nephrology Department of Shanghai Tongji Hospital between July 2022 and May 2023. Patients underwent APT and apparent diffusion coefficient (ADC) mappings within 1 week before biopsy. MRI results of both patients and rats were calculated by comparing with gold standard histology for fibrosis assessment. RESULTS: In animal models, the cortical APT (cAPT) and medullary APT (mAPT) values were positively correlated with the degree of RF. Compared to the sham group, IRI group showed significantly increased cAPT and mAPT values on the 7th and 14th days after surgery, but no group differences were found in ADC values. Similar results were found in human patients. Cortical/medullary APT values were significantly increased in patients with moderate-to-severe fibrosis than in patients with mild fibrosis. ROC curve analysis indicated that APT value displayed a better diagnostic value for RF. Furthermore, combination of cADC and cAPT improved fibrosis detection by imaging variables alone (p < 0.1). CONCLUSION: APT values had better diagnostic capability at early stage of RF compared to ADC values, and the addition of APT imaging to conventional ADC will significantly improve the diagnostic performance for predicting kidney fibrosis.

Layered Composite Dorsal Onlay Implant for Augmentation Rhinoplasty.

The use of autologous costal cartilage in augmentation rhinoplasty is well-established. However, scenarios where costal cartilage is insufficient or patients are unwilling to undergo additional cartilage harvesting present a challenge. This study introduces a composite dorsal onlay implant, combining silicone and costal cartilage, as an effective solution. Twenty female patients were enrolled in this study. Of these patients, 8 underwent revision surgery who had previous rhinoplasty with costal cartilage graft, and 12 had never previously undergone surgery involving the harvesting of costal cartilage. The implant, created by suturing a silicone base with a costal cartilage overlay, demonstrated low rates of warping and translucency over a mean follow-up of 11.4 months. This method offers a refined nasal appearance, particularly a higher dorsum with reduced translucency for patients with limited costal cartilage availability.

Dynamic Phosphoproteomics of BRS3 Activation Reveals the Hippo Signaling Pathway for Cell Migration.

Bombesin receptor subtype-3 (BRS3) is an orphan G-protein coupled receptor (GPCR) that is involved in a variety of pathological and physiological processes, while its biological functions and underlying regulatory mechanisms remain largely unknown. In this study, a quantitative phosphoproteomics approach was employed to comprehensively decipher the signal transductions that occurred upon intracellular BRS3 activation. The lung cancer cell line H1299-BRS3 was treated with MK-5046, an agonist of BRS3, for different durations. Harvested cellular proteins were digested and phosphopeptides were enriched by immobilized titanium (IV) ion affinity chromatography (Ti4+-IMAC) for label-free quantification (LFQ) analysis. A total of 11,938 phosphopeptides were identified, corresponding to 3,430 phosphoproteins and 10,820 phosphosites. Data analysis revealed that 27 phosphopeptides corresponding to six proteins were involved in the Hippo signaling pathway, which was significantly regulated by BRS3 activation. Verification experiments demonstrated that downregulation of the Hippo signaling pathway caused by BRS3 activation could induce the dephosphorylation and nucleus localization of the Yes-associated protein (YAP), and its association with cell migration was further confirmed by kinase inhibition. Our data collectively demonstrate that BRS3 activation contributes to cell migration through downregulation of the Hippo signaling pathway.

Agonist-induced extracellular vesicles contribute to the transfer of functional bombesin receptor-subtype 3 to recipient cells.

Extracellular vesicles (EVs) are important carriers for biomolecules in the microenvironment that greatly promote intercellular and extracellular communications. However, it is unclear whether bombesin receptor-subtype 3 (BRS-3), an orphan G-protein coupled receptor, can be packed into EVs and functionally transferred to recipient cells. In this study, we applied the synthetic agonist and antagonist to activate and inhibit the BRS-3 in HEK293-BRS-3 cells, whose EVs release was BRS-3 activation dependent. The presence of BRS-3 in harvested EVs was further confirmed by an enhanced green fluorescent protein tag. After recipient cells were co-cultured with these EVs, the presence of BRS-3 in the recipient cells was discovered, whose function was experimentally validated. Quantitative proteomics approach was utilized to decipher the proteome of the EVs derived from HEK293-BRS-3 cells after different stimulations. More than 900 proteins were identified, including 51 systematically dysregulated EVs proteins. The Ingenuity Pathway Analysis (IPA) revealed that RhoA signaling pathway was as an essential player for the secretion of EVs. Selective inhibition of RhoA signaling pathway after BRS-3 activation dramatically reversed the increased secretion of EVs. Our data, collectively, demonstrated that EVs contributed to the transfer of functional BRS-3 to the recipient cells, whose secretion was partially regulated by RhoA signaling pathway.

Application of Modified V-Y Advancement Flap in Alar Base Reduction.

BACKGROUND: A variety of techniques for alar base modification have been described in the literature. Limitations of these techniques include scars, recurrence, asymmetry, etc. An incorrect excision of the lateral alar wall can result in a tear-drop or Q-shaped deformity. OBJECTIVES: In this study, a modified V-Y advancement flap will be introduced that will avoid the complications discussed above as well as adjust the positional relationship between the nasal alar and the columella using this technique. In addition, this method is reversible; once the nostril is narrow due to excessive advancement of the flap, the reverse V-Y advancement method can be used to enlarge the nostril. METHODS: In this study, a retrospective review of wide nasal bases in the past 3 years was conducted. A blind analysis of anthropometric points was conducted. There were 3 outcome variables: the width of the alar base, the width of the flare, and the distance between the alar base and columellar base. RESULTS: The alar base was reduced with a modified V-Y advancement flap on 65 female patients ranging from 18 to 45 years of age. In 10 cases, an "Inclined Y-arm flap" was used to narrow alar bases and adjust columella-alar positions. In the remaining cases, a "Horizontal Y-arm flap" was performed to narrow the simple nasal alar width. In all cases, the nasal base was reduced without distortion. In the postoperative period, all the effects were seen to persist for a long time, and patients were generally satisfied with the results of the procedure. CONCLUSION: By using this modified V-Y advancement flap technique, the incision can be concealed as thoroughly as possible, thereby avoiding "tear-drop" or "Q" deformities as well as narrowing the nasal alar effectively. Furthermore, this method, in combination with rhinoplasty surgery, will greatly enhance the nasal appearance.

A Modified Technique in Rhinoplasty: A Septal Extension Graft Complex Using Septal Cartilage, Ethmoid Bone, and Auricular Cartilage.

BACKGROUND: Due to the small and weak septal cartilage of Asians, it is a challenge to obtain ideal tip projection and nose lengthening in this population by performing septal extension grafts with only septal cartilage. OBJECTIVES: The aim of this study was to introduce a septal extension graft complex incorporating nasal septal cartilage, the perpendicular plate of the ethmoid bone, and bilateral auricular cartilage, and to examine its effectiveness in terms of morphological and mechanical support. METHODS: Septal cartilage was harvested under an endoscope according to standard techniques. Two pieces of the perpendicular plate of the ethmoid bone, placed on either side of the "L" strut, served as 2 spreader grafts. A double layer of auricular cartilage was fixed as columellar strut grafts. Three-dimensional facial scanning was used to examine the change of 3 parameters. Nasal tip resistance was evaluated by a digital Newton meter. RESULTS: In total, 25 patients were followed up postoperatively for a mean of 25.9 months. The analysis showed significant differences in both contour parameters and nasal resistance: nasal length (mean [standard deviation], 41.4 [4.0] vs 46.2 [3.4] mm, P < 0.05) and nasal tip projection (21.3 [3.2] vs 24.4 [2.7] mm, P < 0.05) appeared to increase postoperatively, whereas nasolabial angle (117.6° [5.9°] vs 109.5° [5.3°], P < 0.05) showed a significant decrease after surgery. The resistance of the nasal tip also increased significantly (P < 0.05) at displacements of 1, 2, and 3 mm. All patients were satisfied with the aesthetic results, and no serious complications occurred. CONCLUSIONS: This kind of modified septal extension graft technique can effectively adjust nasal morphology for short-nose East Asians.

X-shaped Tip Graft: A Versatile Solution for Warping Correction in Rib Cartilage-Based Rhinoplasty.

BACKGROUND: An enduringly stable tip graft is the key to achieving successful rhinoplasty. However, the intrinsic tendency of rib grafts to warp renders the long-term outcome of cartilage-based rhinoplasty highly unpredictable. OBJECTIVES: The aim of this study was to detail and validate the utilization of a tip graft that is characterized by a circumferential split on the tip graft, which creates a shape similar to the letter X. METHODS: The counterrotating force applied intraoperatively and the complications that arose were examined retrospectively. Three-dimensional stereophotogrammetric evaluations of patients was performed. Anthropometric points were analyzed in a blinded fashion. Outcome variables were tip projection, nasal length, tip rotation, and the nasion-pronasale angulation. RESULTS: Forty-five female patients, ranging in age from 18 to 41 years (average, 28.5 years) completed the study. In all cases, the X graft was applied as an essential element for enhancing tip projection. Postoperative analysis showed excellent maintenance of tip position over time, evidenced by insignificant changes in nasion-pronasale angulation. Other postoperative evaluations, including tip projection, nasal length, and nasolabial angle, showed significant improvement. The differences were not statistically different between short- and long-term follow-up. CONCLUSIONS: The X graft effectively mitigates the negative impact of graft warping. It is the ultimate form for tip support and for elongating short noses. It has the merits of versatility and flexibility when used to create a strong and sustainable tip support in East Asian rhinoplasty.

Analysis of the Cause of Cartilage Warping in the Rhinoplasty of Costal Cartilage and Application of Embed-In Graft in Revisional Surgery.

BACKGROUND: It is undeniable that costal cartilage is an excellent source of grafts, however it has some potential disadvantages, including a tendency to warp. There are very few studies that have examined how to resolve preexisting cartilage warping during secondary revisional surgery. OBJECTIVES: The aim of this study was to explore causes of cartilage warping other than the cartilage itself and ways to correct cartilage warping during revisional surgery. METHODS: In the present study, cases of cartilage warping in the past 5 years were retrospectively reviewed, and external factors of cartilage warping were evaluated to help design new cartilage grafts that could be employed to repair cartilage deformities. RESULTS: A total of 29 females and 3 males between the ages of 18 and 60 years participated in the study (average age, 32.5 years). In all cases, embed-in grafts were utilized to correct warping. All patients were satisfied with the results. As a result of our research, we found that another external cause of cartilage warping was the hump that was not removed completely during the initial surgery. CONCLUSIONS: Incomplete removal of the hump is a major contributing external factor to cartilage warping, particularly in the sagittal plane. This embed-in graft is not intended as a routine graft in normal circumstances, but rather as an immediate means of solving an unexpected situation. The graft is primarily for management of severely warped cartilage.

Continuous Interstitial Glucose Measurement for Flap Venous Occlusion Monitoring in a Diabetic Model.

ABSTRACT: Glucose has been used in the monitoring of transplanted flaps. However, few studies have focused on the effect of diabetes on the decrease in flap glucose levels. The purpose of this study was to evaluate the application of continuous interstitial glucose monitoring to detect flap venous occlusion in a diabetic rat model. The flap interstitial glucose level was measured by transcutaneous sensors in the bilateral vertical rectus abdominis myocutaneous flaps of each rat. interstitial glucose values in the bilateral flaps were recorded every 5 minutes after the venous occlusion of 1 flap. Forty-one rats were divided into the control group (n = 13), diabetic group (n = 15, established by the intraperitoneal injection of streptozotocin), and insulin-treated diabetic group (n = 13, systemic glucose well controlled by insulin before surgery). Results indicated that flap interstitial glucose decreased to the lowest level in all flaps with venous occlusion. The time from venous occlusionto the lowest interstitial glucose level was significantlylongerinthe diabetic group than the control group (120.8 ± 5.7 minutes versus 56.5 ± 6.7 min-minutes, P < 0.001), but was similar in the insulin-treated diabetic and control groups (56.5 ± 6.7 minutes versus 56.0 ± 6.6 minutes, P = 0.876). The change in interstitial glucose in both the diabetic and insulin-treated diabetic groups began with a plateau period or a slow decline before decreasing for 49.0 ± 6.4 minutes and 24.4 ± 4.3 min-minutes, respectively. in a diabetic rat model, impaired glucose metabolism prolongs the required monitoring time and dulls the reaction of flap interstitial glucose to venous congestion. Insulin restores the prolonged monitoring time, but not the reaction of flap interstitial glucose. Therefore, care is needed during application of the technique in diabetic patients.

Psychosocial Experiences in Children With Congenital Melanocytic Nevus on the Face and Their Parents Throughout the Tissue Expansion Treatment.

ABSTRACT: Large congenial melanocytic nevus (LCMN) on the head and face can have negative psychological effects on children and possibly also parents. Patients and their families experience prolonged periods of stress during tissue expansion treatment. The purpose of this study was to evaluate the psychological impact of LCMN on children and parents and explore their fluctuations of psychological status during treatment. Psychosocial status was evaluated in 30 children with LCMN on the head and face and a control group of 56 sex- and age-matched children using the Child Behavior Checklist and the inventory of Subjective Life Quality for Child and Adolescents. The parents of both groups were also evaluated using the Symptom Checklist-90, the Hospital Anxiety and Depression Scale, and the Perceived Stress Scale. Psychological assessments were performed before expander implantation, before lesion resection, and after operation. Results indicated that children with LCMN were more prone to social problems, withdrawal, and aggressive behavior. Furthermore, their psychological problems increased after entering to school. Parents of children with LCMN had high levels of emotional problems and stress. Continuous expansion resulted in the psychosocial status of both children and parents to reach the worst levels during the period before lesion resection. Six months after surgery, most of these psychosocial effects reduced to levels lower than before surgery. Additionally, we identified several risk factors, namely sex, number of operations, being in education, employment status of mother, and family pressures. Large congenial melanocytic nevus on head and face have considerable psychological impact on children and their parents throughout the tissue expansion treatment. This study demonstrates the need for appropriate psychological screening and interventions.

Guided Internal Fixation Technique for Transconjunctival Orbital Fat Transposition.

ABSTRACT: The predominant method for transconjunctival orbital fat transposition is by conducting 1 or several temporary externalized sutures. A new method that could internally and more securely fixate the fat to the periosteum remains desirable. This manuscript introduces a technique that utilizes a titanium ring to guide transconjunctival orbital fat fixation for lower blepharoplasty. With 8 to 10 mm transconjunctival incisions, the orbicularis retaining ligaments were released. An intraoperatively made ring guided the sutures that was made transcutaneously and anchored on the periosteum to exit the preseptal space through the transconjunctival incision. The orbital fat graft was then fixated internally by this suture. From October 2017 to November 2019, 51 patients were included in this study. The mean patient age was 35.7 years (ranged 28-42 years). The mean operative time for the surgical procedure was 52 minutes. No major complications were noticed in any patients. There was 1 case in which the eye bag relapses at fifth week postoperatively, but was corrected by the same approach. At the average follow-up period of 12.1 months, all patients presented significant improvement of the lower eyelid cosmesis. Forty-four patients rated the aesthetic outcomes as "Very Satisfied," the other 7 rated "Satisfied" in a self-assessment questionnaire.Global Aesthetic Improvement Scale echoed similar outcomes. This guided internal fixation technique is technically practical when properly executed. It offers a new method for facilitating orbital fat transposition in lower blepharoplasty.

The Plug-in Graft, the Essential Supplement for a Stable Cartilaginous Framework in Rhinoplasty.

BACKGROUND: A stable cartilaginous framework establishes the foundation for a successful rhinoplasty procedure. However, the curvature of the anterior septal angle usually does not match the added tip graft, leaving a structural gap that can cause postoperative rotation of the nasal tip. A supplementary graft is required to fill this gap, consolidating this structure in 3 dimensions. OBJECTIVES: The aim of this study was to detail and validate the utilization of a graft that fills the structural gap, producing a more stabilized framework and better-sustained tip position in the long term. METHODS: The counterrotating force applied intraoperatively and the complications that arose were collected retrospectively. Three-dimensional stereophotogrammetric evaluations were performed. Anthropometric points were analyzed in a blinded fashion. Outcome variables were tip projection, nasal length, Goode's ratio, and tip rotation. Results were compared statistically. RESULTS: Thirty-one female patients, ranging in age from 18 to 41 years, completed the study. Intraoperative measurements of rotation indicated an unequivocal stabilizing effect of the plug-in graft. Postoperative analysis showed excellent maintenance of the tip position over time. Tip projection and nasolabial angle decreased slightly over time, but the differences were not statistically different. CONCLUSIONS: The plug-in graft evidently consolidates the entire cartilaginous framework in rib-based rhinoplasty procedures. This anatomically integrated structure establishes the fundamental groundwork which provides uncompromising support for the tip graft, thereby achieving an aesthetically pleasing and long-lasting outcome. When executed properly, this graft is effective in maintaining postoperative tip position, justifying the technical challenges involved and the prolonged operating time.

[Discovery of exogenous ligands for orphan receptor BRS-3 from Chinese herbs].

Bombesin receptor subtype-3(BRS-3) is an orphan receptor in the bombesin receptor family. Its signal transduction mechanism and biological function have attracted much attention. Seeking the ligand for BRS-3 is of great significance for exploring its function. Considering the fact that the activation of BRS-3 receptor can induce the change in intracellular Ca~(2+) concentration, the fluo-rometric imaging plate reader(FLIPR) was utilized for ligand screening at the cellular level. Among more than 400 monomeric compounds isolated from Chinese herbs, yuanhunine from Corydalis Rhizoma and sophoraisoflavanone A and licoriphenone from Glycyrrhizae Radix et Rhizoma antagonized BRS-3 to varying degrees. It was confirmed in HEK293 cells expressing BRS-3 that yuanhunine, sophoraisoflavanone A, and licoriphenone inhibited the calcium current response after the activation of BRS-3 by [D-Phe~6,ß-Ala~(11),Phe~(13),Nle~(14)]bombesin-(6-14) in a dose-dependent manner with the IC_(50) values being 8.58, 4.10, and 2.04 µmol·L~(-1), respectively. Further study indicated that yuanhunine and sophoraisoflavanone A exhibited good selectivity for BRS-3. In this study, it was found for the first time that monomers derived from Chinese herbs had antagonistic activity against orphan receptor BRS-3, which has provided a tool for further study of BRS-3 and also the potential lead compounds for new drug discovery. At the same time, it provides reference for the research and development of innovative drugs based on the active ingredients of Chinese herbs.

Comma-Shaped Columellar Strut for Nasal Tip Plasty in East Asian Rhinoplasty.

BACKGROUND: For East Asians, rhinoplasty procedures are predominantly focused on reconstructing a well-defined nasal tip. In this study, the authors present a new graft design for the columellar strut that is similar to the shape of a comma mark. As one integrated piece, the graft undertakes the roles of providing solid support for the tip, modifying different angles, as well as relationships between the tip and columella. METHODS: One hundred and sixty-five female patients, ranging in age from 18 to 42 years (average 27.1 years), received augmentation rhinoplasty surgery over the past 4 years with an average postoperative follow-up of 12 months. Among these patients, 78 of them were primary cases and others were secondary or more. The cartilaginous framework was reconstructed based on comma strut with other grafts including spreaders and dorsal onlay grafts. RESULTS: Patients' own evaluations on the overall improvements of the nose rendered high satisfaction rate. During the long-term follow-up, 8 cases presented asymmetrical nostrils with slightly deviated columella; minor revision surgeries were performed. Another 3 cases presented dorsal graft warping and were also corrected. No other major complications were encountered. CONCLUSIONS: The comma strut provides a reliable support for the nasal tip; its dual curved structure plays the instrumental role in defining the lobular-columellar angle as well as modifying the supratip break. This graft design offers good control of tip and columella; as a result, consistent, reproducible results can be achieved. 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 .

Bilateral Fan-Shaped Septal Extension Struts in East Asian Augmentation Rhinoplasty.

BACKGROUND: For costal cartilage-based rhinoplasty, a method that could properly manage warping of the columellar strut graft and facilitate establishing a stable cartilaginous framework remains desirable. In this study, the authors present a new graft design named bilateral septal extension strut. It utilizes a pair of cartilage grafts with the shape of a fan, which are bilaterally fixated to the septum. Such "sandwiched" structure undertakes the combined functional roles of a columellar strut and a septal extension graft. METHODS: Fifty-two female patients, ranging in age from 18 to 37 years, received augmentation rhinoplasty surgery based on the bilateral septal extension strut. Among these patients, 20 of them were primary cases, 26 secondary, and other six cases were tertiary. RESULTS: Patients' own evaluations on the overall improvements in the nose rendered high satisfaction rate. There was no case of graft extrusion or infections. During the long-term follow-up, one case presented dorsal onlay graft warping and two cases presented in significant asymmetrical nostrils due to the deviation of the columellar at 12 months postoperatively and were corrected in the revision surgery. CONCLUSIONS: The application of the bilateral septal extension struts can help mitigate the negative impact of strut graft warping, making it more manageable. As a result, reliable as well as sustainable support for both tip and lower lateral cartilages can be achieved in the long term. 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 .

Bilateral Tip Add-On Grafts: Concept, Applications and Complications in East Asian Rhinoplasty.

BACKGROUND: A solid tip support plays the fundamental role in augmentation rhinoplasty for East Asians. However, the soft tissue envelope is sometimes limited; the bilateral alae and mucosae cannot be transposed to consolidate the full course of the strut. A graft that serves for the ultimate purpose of elongating the valid length of the strut is therefore necessary. METHODS: Forty-one female patients, ranging in age from 19 to 35 years (average 26.5 years), received augmentation rhinoplasty surgery that was supplemented by the applications of the bilateral accessory add-on tip grafts. The tip projection was established by columellar strut or septal extension grafts. These crescent-shaped grafts acted like a bridge between the alae and the strut, which aim to provide supplementary volume and surface area to help reduce tension and further define the width and shape of the nasal tip. RESULTS: Patients' own evaluations on the overall improvements of the nose rendered high satisfaction rate. During the follow-up, six cases presented deviated columella and asymmetrical nostrils; three cases had deformed dorsal profile due to graft warping or resorption at 12 to 18-month period postoperatively, and were corrected during revision surgeries. No other major complications were encountered. CONCLUSIONS: These accessory tip add-on grafts are intrinsically applicable for East Asians rhinoplasty surgeries. Instead of being utilized as a routine graft for normal circumstances, this pair of accessory tips is an expedient measure to solve a difficult situation. This graft will be particularly necessary in managing a severely short nose with interior soft tissue insufficiency. 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 .

Proteomics Analysis of Cellular BRS3 Receptor Activation Reveals Potential Mechanism for Signal Transduction and Cell Proliferation.

Bombesin-like receptor 3 (BRS3), an orphan G protein-coupled receptor (GPCR), plays important roles in our biological system while the exact mechanisms behind it are less known. To get insights of the biological effects upon BRS3 activation, we utilized quantitative proteomics approach to explore the dynamic protein profiling during the stimulation by its ligand. At different time points after stimulation with BRS3 surrogate agonist, the protein profiling in BRS3 overexpressed HEK 293 cells BRS3 (HEK 293-BRS3) was analyzed by nano-LC-MS/MS. In total, 1593 cellular proteins were confidently identified and quantified, including 146 proteins dysregulated at multiple time points and 319 proteins only altered at one time point. Data analysis indicated that BRS3 activation could regulate cell death, survival, and protein synthesis, particularly mRNA translation. Key signaling pathways were revealed for BRS3 signal transduction. In particular, 21 of our identified proteins are involved in the rapamycin (mTOR) signaling pathway. The promotion of mTOR was further confirmed through monitoring its indicative targets upon BRS3 activation. Upon the inhibition of mTOR by rapamycin, cell proliferation was dramatically reversed. Our proteomics data collectively demonstrate that BRS3 activation will lead to cascades of signal transduction and promote cell proliferation. The developed strategy might be utilized to discover the roles of other GPCRs and improve our understanding of their unknown functions.

Improving the Versatility of the Latissimus Dorsi Myocutaneous Flap Using the Perforator Propeller Flap Concept.

BACKGROUND: The latissimus dorsi myocutaneous flap is widely used in reconstructive surgery; however, primary donor-site closure remains challenging when a wide flap is harvested. METHODS: A large latissimus dorsi myocutaneous flap was elevated and transferred to repair defects. Perforators adjacent to the donor site of the myocutaneous flap were explored using an ultrasound Doppler probe or a technique of extensive exploration along the margins of the donor site wound. A single or multiple perforator propeller flaps based on these perforators were used to close the donor site defect. RESULTS: From June 2012 to April 2018, this method was used to restore posttraumatic and oncologic defects of the chest wall in 14 cases, upper extremity in 6 cases, and lower extremity in 1 case. The size and width of the latissimus dorsi myocutaneous flaps ranged from 16 × 11 cm to 33 × 17 cm (mean area, 335.6 cm) and 9 cm to 20 cm (mean width, 14 cm), respectively. The donor site defect was closed primarily by using a single flap in 11 cases, dual flap in 9, and triple flap in one. Donor site breakdown was not observed in any of the cases. CONCLUSIONS: The perforator propeller flap could be used to reconstruct a latissimus dorsi myocutaneous flap donor site defect, ensuring not only the harvesting of a wide flap but also achieving primary donor site closure, thus greatly improving the versatility and capability of the latissimus dorsi myocutaneous flap in the reconstruction of large-sized defects.

Receptor-specific crosstalk between prostanoid E receptor 3 and bombesin receptor subtype 3.

Bombesin receptor subtype 3 (BRS-3) is a GPCR that is expressed in the CNS, peripheral tissues, and tumors. Our understanding of BRS-3's role in physiology and pathophysiology is limited because its natural ligand is unknown. In an attempt to identify this ligand, we screened toad skin ( Bufo bufo gargarizans Cantor) extracts and identified prostaglandins as putative ligands. In BRS-3-transfected human embryonic kidney (HEK) cells, we found that prostaglandins, with prostaglandin E2 (PGE2) being the most potent, fulfill the pharmacologic criteria of affinity, selectivity, and specificity to be considered as agonists to the BRS-3 receptor. However, PGE2 is unable to activate BRS-3 in different cellular environments. We speculated that EP receptors might be the cause of this cellular selectivity, and we found that EP3 is the receptor primarily responsible for the differential PGE2 effect. Consequently, we reconstituted the HEK environment in Chinese hamster ovary (CHO) cells and found that BRS-3 and EP3 interact to potentiate PGE2 signaling. This potentiating effect is receptor specific, and it occurs only when BRS-3 is paired to EP3. Our study represents an example of functional crosstalk between two distantly related GPCRs and may be of clinical importance for BRS-3-targeted therapies.-Zhang, Y., Liu, Y., Wu, L., Fan, C., Wang, Z., Zhang, X., Alachkar, A., Liang, X., Civelli, O. Receptor-specific crosstalk between prostanoid E receptor 3 and bombesin receptor subtype 3.

Identification of Alkaloids from Corydalis yanhusuo W. T. Wang as Dopamine D1 Receptor Antagonists by Using CRE-Luciferase Reporter Gene Assay.

Corydalis yanhusuo W. T. Wang (C. yanhusuo) has been traditionally used for drug addiction and pain relief in China. In our previous study, we showed that the extract of C. yanhusuo blocks dopamine receptors, demonstrating that its pharmacological activities are mostly due to the antagonistic effects of some of its components at dopamine receptors. As part of our ongoing project on C. yanhusuo, the aim of the present study is to establish a high-throughput and low-cost screening assay system and test the abilities of the isolated alkaloids from C. yanhusuo to inhibit dopamine-induced dopamine D1 receptor activity. By using our established cyclic adenosine monophosphate (cAMP)-response element (CRE)-luciferase reporter gene assay system, we identified eight alkaloids from C. yanhusuo with D1 receptor antagonistic activities. We next validated the activities of these compounds using fluorometric imaging plate reader (FLIPR) assay by measuring the intracellular Ca2+ change. Six out of eight compounds, including tetrahydropalmatine, corydaline, 13-methyldehydrocorydalmine, dehydrocorybubine, dehydrocorydaline, and columbamine, can be confirmed for their inhibitory activities. The dopamine-receptor-antagonistic effects of four compounds, including 13-methyldehydrocorydalmine, dehydrocorydaline, columbamine, and corydaline, are reported for the first time. The present study provides an important pharmacological basis to support the traditional use of C. yanhusuo in China.