ALA-PDT promotes IL-1ß secretion from human SZ95 sebocytes via activation of the NLRP3 inflammasome.

BACKGROUND: 5-Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective treatment for pilosebaceous inflammatory diseases, such as acne vulgaris. In this study, we explored ALA-PDT's mechanisms against acne in vitro. METHODS: We treated human SZ95 sebocytes with ALA (0.2 mM) and subjected them to varied PDT doses (0, 5, 10, 20 J/cm²) over 12 h. We assessed cell viability post-treatment using the Annexin V FITC/PI apoptosis kit. ROS accumulation in the sebocytes was detected with a DCFDA probe. We quantified NLRP3 and caspase-1 mRNA via quantitative PCR and determined IL-1ß release following ALA-PDT by ELISA. Western blotting helped identify the levels of proteins associated with pyroptosis (NLRP3, caspase-1, and IL-1ß). To elucidate the mechanisms, we re-evaluated these parameters after administering various concentrations of NAC antioxidants (0, 0.4, 2, 10 mM) and the caspase inhibitor Z-VAD-FMK (0, 5, 10, 20 µM). RESULTS: Increasing PDT dose inversely affected SZ95 sebocyte survival, with a corresponding rise in ROS and pyroptosis-related proteins (NLRP3, caspase-1, and IL-1ß). Furthermore, NAC and Z-VAD-FMK modulated the expression and secretion of these molecules in a dose-responsive manner. CONCLUSION: Our findings suggest ALA-PDT's potential mechanism of action on sebaceous glands could involve ROS induction, leading to NLRP3 inflammasome assembly, thereby heightening caspase-1 activation and IL-1ß secretion. This cascade may amplify the local inflammatory response to break chronic inflammation in acne vulgaris treatment.

Treatment of linear and whorled nevoid hypermelanosis using QS 694-nm ruby laser.

Laser is being widely used in treating pigmented lesions nowadays. Linear and whorled nevoid hypermelanosis (LWNH) is a rare pigmentary anomaly, and there are only a handful of cases of successful treatment, all with QS 532- and 755-nm laser. The objective of this study was to examine the clinical outcome of QS 694-nm ruby laser in the treatment of LWNH. We report on a 4-year-old boy presented with asymptomatic macular hyperpigmentation over the entire cheek who underwent 3 treatment sessions with QS 694-nm ruby laser. One month after the last treatment, the patient demonstrated significant improvement to the treatment area. Aside from post-procedural purpura lasting approximately 1 week, the patient experienced no serious adverse effects. No recurrence was observed during the 3-month follow-up. Given the excellent results seen in our patients, we recommended the use of QS 694-nm ruby laser as a safe and effective treatment in patients with LWNH.

Ascorbic acid mitigates the deleterious effects of nicotine on tendon stem cells.

Purpose: Nicotine causes tendon degeneration, whereas ascorbic acid imparts beneficial effects on tendon cells. Tendon stem cells (TSCs) play a vital role in maintaining tissue integrity and promoting restoration of structure and function after tendon injury. In the present study, cell culture experiments were performed to determine the effects of nicotine on TSCs and whether ascorbic acid supplementation could antagonize the action of high concentration nicotine. Methods: After treatment with nicotine and ascorbic acid, TSC proliferation, migration, stemness, apoptosis, and differentiation were analyzed. Results: TSC proliferation and expression of stem cell markers were significantly impaired by a high concentration of nicotine (1000 ng/mL), but a lower concentration (100 ng/mL) induced proliferative effects in TSCs. Moreover, the highest concentration of nicotine tested (1000 ng/mL) significantly inhibited the migratory ability of TSCs, while relatively high concentrations (100 and 1000 ng/mL) significantly (p < 0.05) up-regulated non-tenocyte genes. When ascorbic acid was added, the inhibitory effects of nicotine on the proliferation, migration, and stemness of TSCs were reversed. In addition, flow cytometry analysis showed that these nicotine concentrations could induce cell apoptosis, while the addition of ascorbic acid inhibited apoptosis. Conclusion: Addition of ascorbic acid partially reversed the inhibitory effect of a high concentration of nicotine. These findings indicate that while nicotine impairs the biological characteristics of TSCs, ascorbic acid can mitigate these deleterious effects and, therefore, may be useful for decreasing nicotine-induced tendon degeneration.

The preventative effect of bone marrow-derived mesenchymal stem cell exosomes on urethral stricture in rats.

BACKGROUND: Urethral stricture (US) is a major challenge in urology and there is an urgent need for effective therapies for its treatment. Exosomes derived from bone marrow mesenchymal stem cells (BMSCs-Exos) have been shown to be effective in preventing scar and fibrosis formation after tissue injury. However, the potential utility of BMSCs-Exos in the prevention of US remains unknown. We hypothesized that local administration of BMSCs-Exos may influence urethral healing and scar formation in a rat model of US. METHODS: A previously established model of rat US was used in this study. Sprague Dawley rats were randomly assigned into sham, US, and US + BMSCs-Exos groups. Micro-ultrasound assessment, histopathology, immunohistochemistry, and gene expression analysis were performed at four weeks post-surgery. RESULTS: US rats exhibited thick urethral walls with a narrowed lumen, when compared with sham rats. However, these changes were suppressed in the US + BMSCs-Exos group. The preventative effects of BMSCs-Exos on US formation were also apparent histologically. US + BMSCs-Exos rats demonstrated decreased expression of several fibrosis-related genes in urethral tissues, including Col I, fibronectin, and elastin, when compared with US rats. BMSCs-Exos treatment also led to an increase in the expression of angiogenesis-related genes in these tissues, including VEGF, eNOS, and bFGF. CONCLUSIONS: Our findings therefore demonstrate that the local administration of BMSCs-Exos prevents urethral stricture formation by regulating fibrosis and angiogenesis. These findings provide a basis for an innovative strategy involving the clinical application of exosomes to counteract US formation.

Matrix Remodeling-Associated Protein 5 in Urinary Exosomes as a Potential Novel Marker of Obstructive Nephropathy in Children With Ureteropelvic Junction Obstruction.

Recent investigations have described the use of urinary matrix remodeling-associated protein 5 (MXRA5) as a novel biomarker of kidney impairment in the setting of chronic kidney disease. In this study, we aimed to evaluate the possible clinical application of urinary MXRA5 as a useful non-invasive marker in the urine from the affected renal pelvis and bladder of children with ureteropelvic junction obstruction (UPJO). We conducted a prospective cohort study of patients aged <12 months with prenatally diagnosed unilateral UPJO who underwent dismembered pyeloplasty in 2018 or 2019, and a sex- and age-matched control group of healthy children. Blood urea nitrogen and creatinine levels were normal in all the patients. The whole urine and urinary exosomal concentrations of MXRA5 were measured by enzyme-linked immunosorbent assay. The correlations between bladder/renal pelvic MXRA5 levels and differential renal function (DRF) in the affected kidney were also determined. A total of 35 UPJO patients and 12 controls were enrolled in the study. There was no significant difference in whole-urine MXRA5 level between the controls and UPJO patients. However, the exosomal MXRA5 level was significantly lower in the controls than in patients with UPJO (p < 0.05). There were non-significant correlations between bladder and renal pelvis whole-urine MXRA5 levels and DRF (R 2 = 0.1115, p = 0.05 and R 2 = 0.3313, p = 0.0502, respectively). The strongest correlation was between exosomal MXRA5 level in the renal pelvis and DRF (R 2 = 0.8128, p < 0.0001). Urinary exosomal MXRA5 level was significantly higher in children with UPJO than controls. Higher urinary exosomal MXRA5 levels were significantly correlated with lower DRF in the affected kidney in children with UPJO.

"Watch and Wait" Strategy for Multicystic Dysplastic Kidney (MCDK): Status Survey of Perceptions, Attitudes, and Treatment Selection in Chinese Pediatric Urologists and Pediatric Surgeons.

To investigate the perceptions, attitudes, and treatment selection of Chinese pediatric urologists and pediatric surgeons regarding a "watch and wait" strategy for multicystic dysplastic kidney (MCDK). We used a cross-sectional survey in this study. We sent the questionnaire to pediatric urologists and pediatric surgeons to capture their views via the "Questionnaire Star" online survey platform between November and December 2019. The questionnaire contained the basic information and surgical experiences of the respondent, respondents' awareness regarding the counseling of prenatally-diagnosed MCDK and the treatment of MCDK, and respondents' knowledge regarding the imaging modalities, frequency, and duration of follow-up. Of the 200 questionnaires we sent, we received 151 responses. Of those 151 complete responses, most respondents were women (n = 104, 68.9%), pediatric urologists (n = 78, 51.6%), and practicing with at least 5 years of surgical experience (n = 112, 74.2%); 11.9% reported >20 years' experience. Eighty-two surgeons (54.3%) provided positive counseling for prenatally-diagnosed MCDK. Ninety-nine surgeons (65.6%) advocated conservative management for MCDK, and only 14.8% of respondents suggested limiting the use of radiographic evaluation for MCDK. Surgeons working in academic teaching facilities and those from East China were more likely to select a "watch and wait" strategy. Chinese pediatric urologists and pediatric surgeons have inadequate knowledge of the "watch and wait" strategy for MCDK. An expert consensus on the strategy of "watch and wait" for MCDK in China is urgently needed to promote the application of this non-surgical treatment mode in clinical practice. A larger sample size is required to fully identify the current opinion of Chinese pediatric urologists and pediatric surgeons regarding the management of MCDK.

Extracellular vesicles produced by bone marrow mesenchymal stem cells attenuate renal fibrosis, in part by inhibiting the RhoA/ROCK pathway, in a UUO rat model.

BACKGROUND: Extracellular vesicles produced by bone marrow mesenchymal stem cells (BMSC-EVs) can play important roles in the repair of injured tissues. Though numerous studies have reported the effect of EVs on renal fibrosis, the underlying mechanisms remain unclear. We hypothesized that BMSC-EVs containing milk fat globule-epidermal growth factor-factor 8 (MFG-E8) could attenuate renal fibrosis by inhibiting the RhoA/ROCK pathway. METHODS: We investigated whether BMSC-EVs have anti-fibrotic effects in a rat model of renal fibrosis, in which rats were subjected to unilateral ureteral obstruction (UUO), as well as in cultured HK2 cells. Extracellular vesicles from BMSCs were collected and co-cultured with HK2 cells during transforming growth factor-ß1 (TGF-ß1) treatment. HK2 cells co-cultured with TGF-ß1 were also treated with the ROCK inhibitor, Y-27632. RESULTS: Compared with the Sham group, UUO rats displayed fibrotic abnormalities, accompanied by an increased expression of α-smooth muscle actin and Fibronectin and reduced expression of E-cadherin. These molecular and pathological changes suggested increased inflammation in damaged kidneys. Oxidative stress, as evidenced by an increased level of MDA and decreased levels of SOD1 and Catalase, was also observed in UUO kidneys. Additionally, activation of cleaved caspase-3 and PARP1 and increased apoptosis in the proximal tubules confirmed tubular cell apoptosis in the UUO group. All of these phenotypes exhibited by UUO rats were suppressed by treatment with BMSC-EVs. However, the protective effect of BMSC-EVs was completely abolished by the inhibition of MFG-E8. Consistent with the in vivo results, treatment with BMSC-EVs reduced inflammation, oxidative stress, apoptosis, and fibrosis in HK-2 cells stimulated with TGF-ß1 in vitro. Interestingly, treatment with Y-27632 protected HK-2 cells against inflammation and fibrosis, although oxidative stress and apoptosis were unchanged. CONCLUSIONS: Our results show that BMSC-EVs containing MFG-E8 attenuate renal fibrosis in a rat model of renal fibrosis, partly through RhoA/ROCK pathway inhibition.

Congenital Anomalies of the Kidney and Urinary Tract in Children with Congenital Heart Defects.

BACKGROUND/AIMS: To investigate the incidence and clinical characteristics of congenital anomalies of the kidney and urinary tract (CAKUT) in children with congenital heart defects (CHD). METHODS: We retrospectively analyzed the clinical data of children with CHD with CAKUT admitted to the Shanghai Children's Medical Center affiliated with the Shanghai Jiao Tong University School of Medicine between September 2018 and March 2019. Patients underwent routine examinations for liver, kidney, and coagulation function, and urinary tract ultrasonography, and we summarized patients' clinical manifestations and imaging abnormalities. RESULTS: A total of 1,410 children with CHD were diagnosed and treated in our hospital. The total number of patients with abnormal urogenital systems was 104, and hydronephrosis was the most common abnormality, followed by vesicoureteral reflux and duplication of the kidney and ureter. The overall prevalence of CAKUT was 7.4%. There was no statistically significant difference for maternal age, sex, parity, gestational age, and history of medication during pregnancy between the patients with CAKUT and those without CAKUT. CONCLUSION: The incidence of CAKUT in our patients with CHD was significantly higher than that in the general population. We recommend urinary ultrasonography as a routine examination for children with CHD for early detection of CAKUT, to avoid missed diagnoses, and to initiate appropriate treatment.

MFG-E8 regulates inflammation and apoptosis in tendon healing, and promotes tendon repair: A histological and biochemical evaluation.

Several studies have identified potential roles for MFG-E8 in promoting tissue repair. However, the effects of MFG-E8 on tendon repair have not yet been investigated. Therefore, we explored the role of MFG-E8 on tendon repair using a rat model of patellar tendon injury. The patellar tendons of Sprague Dawley rats (n = 24/group) received window defects and, after modeling, three groups were randomly assigned: (a) recombinant MFG-E8 (rMFG-E8) group, implantation with MFG-E8 and fibrin glue (400 ng in 10 µl); (b) fibrin group, implantation with fibrin only; and (c) control group, without any treatment. Histopathology, immunohistochemistry, and gene expression analyses were performed at 2 and 4 weeks after healing. Administration of rMFG-E8 in injury sites significantly improved tendon healing histologically at 4 weeks after injury. In addition, numbers of M1 macrophages and M1-stimulator genes, including IFNG, Il-1B, and Il-6, were reduced in the repair sites at 2 weeks by rMFG-E8 administration. In parallel, rMFG-E8 significantly increased the number of M2 macrophages and expression of anti-inflammatory IL-10 and IL-4 at 2 weeks after injury. Treatment with rMFG-E8 markedly decreased tendon cell apoptosis. Moreover, rMFG-E8 significantly enhanced the expression of genes related to tendon matrix formation at 2 weeks after injury, including Col1a1and tenascin-C. We conclude that MFG-E8 could regulate inflammatory responses and apoptotic cell accumulation in tendon repair, and promote the healing process of injured tendon tissue. Thus, exogenous application of MFG-E8 might have therapeutic potential for repair of tendon injuries.

Extracellular vesicles from bone marrow-derived multipotent mesenchymal stromal cells regulate inflammation and enhance tendon healing.

BACKGROUND: Extracellular vesicles from bone marrow-derived multipotent mesenchymal stromal cells (BMSC-EVs) can play important roles in the repair of injured tissues. However, no reports have investigated the role and underlying mechanisms of BMSCs-EVs in the tendon repair process. We hypothesized that BMSC-EVs may play a role in modulating inflammation during tendon healing and improving tendon repair in a rat model of patellar tendon injury. METHODS: First, we created window defects in the patellar tendons of Sprague-Dawley rats. Rats (n = 16) were then randomly assigned to three groups: BMSC-EVs group, Fibrin group, and control group. Rats in the BMSC-EVs group were treated with BMSC-EVs and fibrin glue (25 µg in 10 µL). Rats in the fibrin group were treated with fibrin only, and those in the control group received no treatment. Histopathology, immunohistochemistry, and gene expression analyses were performed at 2 and 4 weeks after surgery. RESULTS: At 4 weeks, tendons treated with BMSC-EVs showed regularly aligned and compact collagen fibers as compared with the disrupted scar-like healing in rats in the fibrin and control groups. The expression of genes related to tendon matrix formation and tenogenic differentiation: collagen (COL)-1a1, scleraxis (SCX), and tenomodulin (TNMD) was significantly higher in the BMSC-EVs group than in the other two groups. With histopathology, we observed significantly higher numbers of CD146+ tendon stem cells and fewer numbers of apoptotic cells and C-C chemokine receptor type 7 (CCR7)-positive proinflammatory macrophages in the BMSC-EVs group. BMSC-EVs treatment also led to an increase in the expression of anti-inflammatory mediators (IL-10 and IL-4) at 2 weeks after surgery. CONCLUSIONS: Overall, our findings show that the local administration of BMSC-EVs promotes tendon healing by suppressing inflammation and apoptotic cell accumulation and increasing the proportion of tendon-resident stem/progenitor cells. These findings provide a basis for the potential clinical use of BMSC-EVs in tendon repair.

Congenital Stenosis of the External Orifice of the Urethra in a Female Child.

Limited numbers of pediatric stenosis of the external orifice of the urethra have been reported. We report a case of congenital stenosis of the external orifice of the urethra in a female child who underwent meatoplasty. As an initial strategy for congenital stenosis of the external orifice of the urethra in girls, dilatation of the stenosed urethral meatus may be another management of choice.