Epithelial Membrane Research Articles

Insecticidal, genotoxic and biochemical effects of Shewanella sp. (SS4) and Thauera sp. (M9) on Spodoptera litura

AbstractSpodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is a polyphagous pest causing economic losses to a number of agricultural crops. The present study was conducted to explore the insecticidal potential of some soil bacteria against S. litura. Among the tested bacterial isolates, Shewanella sp. (SS4) and Thauera sp. (M9) caused higher larval mortality. Concentration–response bioassays indicated a dose‐dependent increase in larval mortality with maximum mortality of 80 and 76%, respectively, due to Shewanella sp. and Thauera sp. Both the bacteria negatively affected the growth and development, reduced adult emergence and induced morphological deformities in adults. Nutritional parameters of S. litura i.e. relative growth and consumption rate, efficiency of conversion of ingested and digested food as well as approximate digestibility of food decreased with increasing bacterial concentrations. Histopathological studies revealed the disruption of peritrophic membrane and midgut epithelium of larvae. Profiles of digestive enzymes i.e. α‐amylases, α, β‐glucosidases and galactosidases, lipases and proteases were also significantly decreased with bacterial treatment. Comet assay test indicated DNA damage in larval haemocytes due to bacterial infection. These results imply that both these bacterial isolates have the potential as biological control agents and may prove as environmentally friendly alternative for pest management techniques.

Reaction of Kidney Medullary Laminin to Prolonged Dehydration of Rats

Laminin is the main water-soluble protein in the basement membrane of epithelial tissue. The content of laminin in the renal parenchyma was studied in rats that were on an alternative water diet for 60 h with excessive water intake or a complete lack of drinking water in the food. It has been shown that the osmotic concentration of urine, which develops as a result of water deprivation, is accompanied by a quantitative change in the composition of laminin. It has been established that the amount of laminin beta chain increases in the kidney medulla of dehydrated rats. This effect is observed only in WAG rats with normally expressed vasopressin gene and is absent in mutant Brattleboro rats unable to synthesize the hormone. The increase in the level of beta-laminin also does not extend to the cortical substance. Based on the original and new data, it is assumed that laminin, as a key regulatory element in the basement membrane of the tubular epithelium, participates in the adaptive reaction of the concentrating system of the kidney to conditions of prolonged dehydration. Hyperhydration does not affect the level of laminin in the renal parenchyma.

Mooren’s Ulcer with Pathologic Findings Similar to Cicatricial Pemphigoid

Abstract To report a case of peripheral corneal thinning with a presentation similar to a Mooren’s ulcer but with pathology consistent with cicatricial pemphigoid. We report the case of a 56-year-old woman who presented with unilateral peripheral corneal thinning. Before presentation, the patient was treated with prednisone 60 mg daily, an amniotic membrane, and artificial tears. The patient denied any prior autoimmune history. The left cornea had nasal crescentic corneal thinning 6.0 mm tall and 1.5 mm wide. Visual acuity was 20/40 uncorrected in the involved eye. Initial laboratory workup was notable for a low positive rheumatoid factor and positive fluorescent treponemal antibody absorption but negative rapid plasma reagin. Corneal cultures were negative. The patient was started on oral and topical steroids, topical moxifloxacin, and mycophenolate mofetil and underwent a conjunctival resection. The conjunctival tissue was sent for pathology and was notable for strong deposits of immunoglobulin G at the epithelial basement membrane consistent with mucous membrane pemphigoid. Mycophenolate mofetil was discontinued due to oral thrush. Due to persistent activity, she underwent repeat conjunctival resection with the placement of an amniotic membrane over the area of thinning. She was subsequently started on rituximab biosimilar infusions and tapered off prednisone. Visual acuity at the last follow-up was 20/25. While conjunctival resection is a potential management option for Mooren’s ulcer, routine conjunctival biopsy for pathology is not. Our case highlights a potential overlap between the pathophysiology of ocular cicatricial pemphigoid and Mooren’s ulcer. Sending conjunctival tissue for pathology could provide benefits to the underlying mechanisms of different variants of Mooren’s ulcer.

Aplasia Cutis Congenita With Cutaneous Meningioma: A Rare Case.

Cutaneous meningioma, characterized by ectopic meningothelial cells in the dermis or subcutis, is a rare neoplasm. Generally, the most common location for cutaneous meningioma is the scalp. We report a case of cutaneous meningioma presenting as soft, light red, atrophic, and smooth patches with central blue spots. On histological examination, the tumor consisted largely of epithelioid cells, whorls, nests, and syncytial sheets of meningothelial cells. HMB-45, Melan-A, and S100 were negative; epithelial membrane antigen and somatostatin receptor 2 were positive. Ultimately, histopathologic examination and immunohistochemistry results supported a diagnosis of cutaneous meningioma. In addition, dermal dysplasia was observed above the tumor, manifested by thinning of the dermis and loss of appendages. No abnormalities were found on brain magnetic resonance imaging. Cutaneous meningioma is an extremely rare tumor, and its manifestation as an atrophic patch is even rarer. There have been mainly clinical reports of cutaneous meningioma. This was a rare case of focal aplasia cutis congenita with cutaneous meningioma. For cutaneous meningioma to be diagnosed earlier, there needs to be greater public awareness about the condition.

Branched-chain actin dynamics polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains.

In prevailing epithelial polarity models, membrane- and junction-based polarity cues such as the partitioning-defective PARs specify the positions of apicobasal membrane domains. Recent findings indicate, however, that intracellular vesicular trafficking can determine the position of the apical domain, upstream of membrane-based polarity cues. These findings raise the question of how vesicular trafficking becomes polarized independent of apicobasal target membrane domains. Here, we show that the apical directionality of vesicle trajectories depends on actin dynamics during de novo polarized membrane biogenesis in the C. elegans intestine. We find that actin, powered by branched-chain actin modulators, determines the polarized distribution of apical membrane components, PARs, and itself. Using photomodulation, we demonstrate that F-actin travels through the cytoplasm and along the cortex toward the future apical domain. Our findings support an alternative polarity model where actin-directed trafficking asymmetrically inserts the nascent apical domain into the growing epithelial membrane to partition apicobasal membrane domains.

The biosynthetic-secretory pathway, supplemented by recycling routes, determines epithelial membrane polarity.

In prevailing epithelial polarity models, membrane-based polarity cues (e.g., the partitioning-defective PARs) position apicobasal cellular membrane domains. Intracellular vesicular trafficking expands these domains by sorting polarized cargo toward them. How the polarity cues themselves are polarized in epithelia and how sorting confers long-range apicobasal directionality to vesicles is still unclear. Here, a systems-based approach using two-tiered C. elegans genomics-genetics screens identifies trafficking molecules that are not implicated in apical sorting yet polarize apical membrane and PAR complex components. Live tracking of polarized membrane biogenesis indicates that the biosynthetic-secretory pathway, linked to recycling routes, is asymmetrically oriented toward the apical domain during this domain's biosynthesis, and that this directionality is regulated upstream of PARs and independent of polarized target membrane domains. This alternative mode of membrane polarization could offer solutions to open questions in current models of epithelial polarity and polarized trafficking.

SMYD3 drives the proliferation in gastric cancer cells via reducing EMP1 expression in an H4K20me3-dependent manner

Protein lysine methyltransferase SET and MYND domain-containing 3 (SMYD3) is aberrantly expressed in various cancer settings. The mechanisms that SMYD3 activates the expression of critical pro-tumoral genes in an H3K4me3-dependent manner have been well described in previous reports. Besides H3K4me3, H4K20me3 is another catalytic product of SMYD3, however it is a transcriptionally repressive hallmark. Since it is not clear that how SMYD3-elicited transcriptionally repressive program functions in cancer, we used gastric cancer (GC) as a model to investigate the roles of SMYD3-H4K20me3. Herein, online bioinformatics tools, quantitative PCR, western blotting and immunohistochemistry assays demonstrated that SMYD3 expression was markedly increased in GC tissues from our institutional and The Cancer Genome Atlas (TCGA) cohort. Additionally, aberrantly increased SMYD3 expression was closely associated with aggressive clinical characteristics and poor prognosis. Depletion of endogenous SMYD3 expression using shRNAs significantly attenuates the proliferation in GC cells and Akt signaling pathway in vitro and in vivo. Mechanistically, chromatin immunoprecipitation (ChIP) assay showed that SMYD3 epigenetically repressed the expression of epithelial membrane protein 1 (EMP1) in an H4K20me3-dependent manner. Gain-of-function and rescue experiments validated that EMP1 inhibited the propagation of GC cells and reduced p-Akt (S473) level. Based on these data, pharmaceutical inhibition of SMYD3 activity using the small inhibitor BCI-121 deactivated Akt signaling pathway in GC cells and further impaired the cellular viability in vitro and in vivo. Together, these results demonstrate that SMYD3 promotes the proliferation in GC cells and may be a valid target for therapeutic intervention of patients with GC.

Transforming growth factor-β1 and SMAD signalling pathway in the small airways of smokers and patients with COPD: potential role in driving fibrotic type-2 epithelial mesenchymal transition.

COPD is a common disease characterized by respiratory airflow obstruction. TGF-β1 and SMAD pathway is believed to play a role in COPD pathogenesis by driving epithelial mesenchymal transition (EMT). We investigated TGF-β1 signalling and pSmad2/3 and Smad7 activity in resected small airway tissue from patients with; normal lung function and a smoking history (NLFS), current smokers and ex-smokers with COPD GOLD stage 1 and 2 (COPD-CS and COPD-ES) and compared these with normal non-smoking controls (NC). Using immunohistochemistry, we measured activity for these markers in the epithelium, basal epithelium, and reticular basement membrane (RBM). Tissue was also stained for EMT markers E-cadherin, S100A4 and vimentin. The Staining of pSMAD2/3 was significantly increased in the epithelium, and RBM of all COPD groups compared to NC (p <0.0005). There was a less significant increase in COPD-ES basal cell numbers compared to NC (p= 0.02). SMAD7 staining showed a similar pattern (p <0.0001). All COPD group levels of TGF-β1 in the epithelium, basal cells, and RBM cells were significantly lower than NC (p <0.0001). Ratio analysis showed a disproportionate increase in SMAD7 levels compared to pSMAD2/3 in NLFS, COPD-CS and COPD-ES. pSMAD negatively correlated with small airway calibre (FEF25-75%; p= 0.03 r= -0.36). EMT markers were active in the small airway epithelium of all the pathological groups compared to patients with COPD. Activation of the SMAD pathway via pSMAD2/3 is triggered by smoking and active in patients with mild to moderate COPD. These changes correlated to decline in lung function. Activation of the SMADs in the small airways is independent of TGF-β1, suggesting factors other than TGF-β1 are driving these pathways. These factors may have implications for small airway pathology in smokers and COPD through the process of EMT, however more mechanistic work is needed to prove these correlations.

DP18 Extraocular sebaceous carcinoma in situ

Abstract Extraocular sebaceous carcinoma in situ (SCIS) is exceedingly rare with &amp;lt; 10 cases reported in the literature (Jeong KM, Seo JY, Kim A et al. Extraocular sebaceous carcinoma in situ concurrent with actinic keratosis: a case report and review of the literature. Korean J Dermatol 2021; 59:307–9). An 80-year-old man reported a 3-month history of a right forehead lesion that had been bleeding. This was a subtle 5 × 5 mm papule on clinical examination, and dermoscopy showed arborizing telangiectasia. Past medical history was significant for 20 basal cell carcinomas, one squamous cell carcinoma, Bowen disease and actinic keratoses. Histology showed sun-damaged skin with a partially eroded neoplasm of lobular architecture, with surface and infundibular epithelium containing atypical sebocytes and squamous cells with no infiltrative growth pattern, which was consistent with SCIS and was fully excised. Immunohistochemistry revealed patchy adipophilin and androgen receptor staining and stronger epithelial membrane antigen staining. Furthermore, there was preserved expression of MSH2, MSH6, MLH1 and PMS2. Sebaceous carcinoma in situ is a debated entity not formally recognized in the classification of sebaceous neoplasms. Kazakov et al. reported five cases of dermal sebaceous tumours with ‘benign’ architectural features (well-circumscribed and no evidence of invasion) but with cytological atypia (abnormal mitoses and pleomorphism) (Kazakov D, Kutzner H, Spagnolo DV et al. What is extraocular cutaneous sebaceous carcinoma in situ? Am J Dermatopathol 2010; 32:857–8). The authors questioned whether these cases represented sebaceous adenomas with atypia or sebaceous carcinomas (Kazakov et al.). In response, it has been argued that such architectural–cytological discordant tumours can be regarded as SCIS owing to the fact that, by definition, carcinoma in situ is a cancer that remains wholly at the site of origin. However, it is argued that these lesions have an unclear origin and may not arise from sebaceous glands in the skin, unlike the more common aggressive periorbital sebaceous carcinomas that arise from meibomian glands and glands of Zeis (Kazakov et al.). It has been postulated that a SCIS diagnosis should be reserved for intraepidermal tumours not extending beyond the epidermal basement membrane, which is a vanishing minority of sebaceous neoplasms (Kazakov et al.). Similar to our case, a 2021 case report identified SCIS without any connection to an adnexal component but concurrent with actinic keratosis. These authors proposed that the tumour cells of intraepidermal squamous neoplasia can differentiate towards malignant sebocytes (Jeong et al.). Although controversial, SCIS appears to have reasonable, clear histological criteria for diagnosis and an excellent prognosis, with no reports of recurrence following surgical excision. We feel it is important to increase identification to avoid the overtreatment of such lesions.

CPC12 Coinheritance of BRCA2 and CYLD germline pathogenic variants associated with targetable metastatic malignant cylindroma

Abstract CYLD cutaneous syndrome (CCS) presents during puberty with the development of hair follicle tumours, including cylindromas, spiradenomas or trichoepitheliomas, and is associated with pathogenic variants in the tumour suppressor gene CYLD. Germline pathogenic variants of BRCA2 are associated with an increased risk of breast, ovarian, pancreatic and prostate cancer. Coinheritance of pathogenic variants in both genes has not been previously reported, and here, we present a 29-year-old man with metastatic malignant cylindroma, where knowledge of the germline variants has targetable therapeutic implications. The proband presented with cylindromas on his scalp from the age of 8 years, inherited from his mother who had CCS. The proband’s father and paternal grandmother both had early-onset breast cancer. The proband’s 33-year-old sister had been diagnosed with breast cancer and CCS; genetic testing revealed germline pathogenic variants in both BRCA2 and CYLD. Aged 28 years, the proband presented with a growing, ulcerated cylindroma on the scalp. Histology revealed a biphasic neoplasm, showing malignant cylindroma with high-grade features arising within a pre-existing benign cylindroma. The malignant epithelioid component of the tumour showed brisk mitotic activity (25 mitoses per mm2) and large infiltrative nests with central comedo-type necrosis. Immunohistochemistry showed positivity for epithelial membrane antigen, CK7, p63 and patches of CK5/6, whereas carcinoembryonic antigen, S100 and human melanoma black-45 were negative. Several weeks after excision of the malignant scalp cylindroma, a lump presented on the left side of the patient’s neck; biopsy revealed histology consistent with the malignant scalp cylindroma. Radiological imaging revealed metastatic deposits of malignant cylindroma in the right axilla, both lungs and the brain. Genetic testing of the proband revealed germline pathogenic variants in both BRCA2 and CYLD, as seen in his sister. The patient received radiotherapy for the tumour in his neck, with partial resolution of symptoms and gamma-knife ablation of brain metastases. DNA extracted from the patient’s tumour was analysed using a high-resolution single-nucleotide polymorphism array, which demonstrated loss of heterozygosity including the BRCA2 and CYLD loci. The germline and somatic pathogenic variants in this patient support the use of unlicensed therapeutics that target BRCA deficiency, such as poly-ADP ribose polymerase inhibitors, which have not previously been used to treat malignant cylindroma. This case emphasizes the importance of recognizing coinheritance of tumour suppressor genes in rare genetic conditions, as this can provide novel management options and improve patient outcomes.

Research Progress on the Mechanism of Nanoparticles Crossing the Intestinal Epithelial Cell Membrane.

Improving the stability of drugs in the gastrointestinal tract and their penetration ability in the mucosal layer by implementing a nanoparticle delivery strategy is currently a research focus in the pharmaceutical field. However, for most drugs, nanoparticles failed in enhancing their oral absorption on a large scale (4 folds or above), which hinders their clinical application. Recently, several researchers have proved that the intestinal epithelial cell membrane crossing behaviors of nanoparticles deeply influenced their oral absorption, and relevant reviews were rare. In this paper, we systematically review the behaviors of nanoparticles in the intestinal epithelial cell membrane and mainly focus on their intracellular mechanism. The three key complex intracellular processes of nanoparticles are described: uptake by intestinal epithelial cells on the apical side, intracellular transport and basal side exocytosis. We believe that this review will help scientists understand the in vivo performance of nanoparticles in the intestinal epithelial cell membrane and assist in the design of novel strategies for further improving the bioavailability of nanoparticles.

Hepatic myxosarcoma in a domestic shorthair cat.

Myxosarcomas are rare malignant neoplasms of soft connective tissues, and there are no reports of hepatic myxosarcomas in cats. An eight-year-old male, neutered, domestic shorthair cat presented with progressive hyporexia, lethargy, and weight loss. An ultrasonography study showed a large abdominal mass connected to the liver. The cat underwent a laparotomy and the mass was removed. Histopathological evaluation of the mass supported the diagnosis of a myxosarcoma. Tumour cells were positive with vimentin and alcian blue stain, and negative with PAS, pan-cytokeratin, s100, epithelial membrane antigen, and α-smooth muscle actin. The Ki-67 index by immunohistochemistry was 6%. The cat was euthanased due to severe lethargy and recumbency. Myxoid soft tissue neoplasms are very rare in cats, and to the best of our knowledge, this is the first report of a hepatic myxosarcoma in a cat. In the present case, the diagnosis was made based on histopathological and immunohistochemical findings and an alcian blue-positive supporting matrix.

Self-Assembled Sulfated Hyaluronan Coating Modulates Transforming Growth Factor-Beta1 Penetration for Corneal Scarring Alleviation.

Corneal scarring caused by epithelial-stromal injury impairs corneal transparency and visual acuity. Excess secretion of transforming growth factor-beta 1 (TGF-β1), which promotes wound closure, penetrates the corneal stroma via defects in the epithelial basement membrane and induces the differentiation of corneal fibroblasts to myofibroblasts, leading to scar formation. Modulating TGF-β1 penetration might alleviate corneal scar formation and restore transparency. In this study, sulfated hyaluronan (sHA) coatings were self-assembled above wounded corneal stroma to modulate TGF-β1 penetration, and their ability to alleviate corneal scarring was investigated. The formation of sHA coatings was rapid (within 30 s), and the high-sulfated hyaluronan coating efficiently blocked penetration by TGF-β1 and reduced the concentration of TGF-β1 in the corneal stroma. Further investigation showed that the ability of TGF-β1 to induce differentiation of corneal fibroblasts into myofibroblasts was inhibited by sHA binding. Evaluation of corneal scarring with sHA coating in a rabbit model of lamellar resection indicated that a sHA (high sulfation) coating effectively reduced scar formation. Immunohistochemical staining of α-smooth muscle actin and optical coherence tomography of the anterior segment showed minimal scar tissue formation in the sHA group. This work presents a promising alternative to alleviate scarring in corneal epithelial-stromal injury.

Coelenterate and Germinal-Müllerian Papilloma Cervix: A Mini-Review

Coelenterate and Germinal-Müllerian Papilloma Cervix: A Mini-Review

Upregulation of SEMP1 Contributes to Improving the Biological Functions of Trophoblast via the PI3K/AKT Pathway in Preeclampsia.

Disturbance of extravillous trophoblast infiltration is associated with preeclampsia (PE), a severe condition of pregnancy characterized by hypertension and proteinuria. Senescence-associated epithelial membrane protein 1 (SEMP1), an integral membrane protein, is a vital component of tight junction strands in epithelial or endothelial cells, with no clear function reported in PE. Gene Expression Omnibus (GEO) datasets showed that SEMP1 expression was downregulated in the placental tissues of PE patients, which was confirmed by assessing SEMP1 levels in placental samples collected in our hospital. Furthermore, less SEMP1 was detected in cytokeratin 7 positive trophoblast cells in the spiral arteries of rat placentas post L-arginine methyl ester hydrochloride (L-NAME) treatment. Trophoblast cells acquired robust ability of proliferation, migration, and invasion when SEMP1 was overexpressed. Such capability was weakened in SEMP1-silenced cells. Trophoblast cells overexpressing SEMP1 secreted more vascular endothelial growth factor A (VEGFA), which facilitated the tube formation of human umbilical vein endothelial cells. Blockade of PI3K/AKT signaling transduction with LY294002 dampened the effects of SEMP1 on trophoblast cells. Collectively, we firstly indicated that SEMP1 inhibition is a potential driver for PE, which may be associated with the deactivation of the PI3K/AKT pathway.

Needle tract seeding of a sclerosing epithelioid fibrosarcoma in a biopsy tract: a case report

BackgroundA sclerosing epithelioid fibrosarcoma (SEF) is an uncommon tumor of the deep soft tissue. An SEF has been described as a low-grade tumor with high local recurrence and metastatic rates. Generally, in bone and soft tissue tumors, a resection of the biopsy route is recommended; however, there is limited evidence with respect to the dissemination of the tumor tissue during a needle biopsy.Case presentationA mass in the right pelvic cavity, with no symptoms, was observed in a 45-year-old woman during a gynecological examination. Computed tomography (CT) revealed a multilocular mass with calcification in the pelvic cavity. The magnetic resonance imaging (MRI) showed an iso-signal intensity on T1 weighted images and hypo- and iso-signal intensity on T2 weighted images. The CT-guided core needle biopsy was performed using a dorsal approach, and the biopsy diagnosis was a low-grade spindle cell tumor. The tumor was excised using an anterior approach. The tumor tissue comprised spindle cells and epithelioid cells with irregular nuclei, and the immunohistological analysis was positive for vimentin and epithelial membrane antigen, which was consistent with a diagnosis of sclerosing epithelioid fibrosarcoma. Five years after the surgery, the MRI showed a tumor recurrence in the subcutaneous tissue of the right buttock, which was consistent with the needle biopsy tract. The patient underwent a tumor excision, and the resected tumor was similar to the primary tumor.ConclusionsThe recurrent tumor was excised with a surgical margin, and the tumor specimen had the histological features of a sclerosing epithelioid fibrosarcoma. It was difficult to investigate the association of the core needle biopsy with the tumor recurrence because the approach of the biopsy tract is usually same as that used in a tumor excision. However, the present case indicated the tumor may recur in the biopsy tract of a soft tissue sarcoma. Surgeons should be aware of the possibility of disseminating tumor tissues in a needle biopsy.

The impact of UV cross-linking on corneal stromal cell migration, differentiation and patterning

Previous studies have demonstrated that UV cross-linking (CXL) increases stromal stiffness and produces alterations in extracellular matrix (ECM) microstructure. In order to investigate how CXL impacts both keratocyte differentiation and patterning within the stroma, and fibroblast migration and myofibroblast differentiation on top of the stroma, we combined CXL with superficial phototherapeutic keratectomy (PTK) in a rabbit model. Twenty-six rabbits underwent a 6 mm diameter, 70 μm deep phototherapeutic keratectomy (PTK) with an excimer laser to remove the epithelium and anterior basement membrane. In 14 rabbits, standard CXL was performed in the same eye immediately after PTK. Contralateral eyes served as controls. In vivo confocal microscopy through focusing (CMTF) was used to analyze corneal epithelial and stromal thickness, as well as stromal keratocyte activation and corneal haze. CMTF scans were collected pre-operatively, and from 7 to 120 days after the procedure. A subset of rabbits was sacrificed at each time point, and corneas were fixed and labeled in situ for multiphoton fluorescence microscopy and second harmonic generation imaging. In vivo and in situ imaging demonstrated that haze after PTK was primarily derived from a layer of myofibroblasts that formed on top of the native stroma. Over time, this fibrotic layer was remodeled into more transparent stromal lamellae, and quiescent cells replaced myofibroblasts. Migrating cells within the native stroma underneath the photoablated area were elongated, co-aligned with collagen, and lacked stress fibers. In contrast, following PTK + CXL, haze was derived primarily from highly reflective necrotic “ghost cells” in the anterior stroma, and fibrosis on top of the photoablated stroma was not observed at any time point evaluated. Cells formed clusters as they migrated into the cross-linked stromal tissue and expressed stress fibers; some cells at the edge of the CXL area also expressed α-SM actin, suggesting myofibroblast transformation. Stromal thickness increased significantly between 21 and 90 days after PTK + CXL (P < 0.001) and was over 35 μm higher than baseline at Day 90 (P < 0.05). Overall, these data suggest that cross-linking inhibits interlamellar cell movement, and that these changes lead to a disruption of normal keratocyte patterning and increased activation during stromal repopulation. Interestingly, CXL also prevents PTK-induced fibrosis on top of the stroma, and results in long term increases in stromal thickness in the rabbit model.

Kidney targeting peptide-modified biomimetic nanoplatforms for treatment of acute kidney injury.

The management of acute kidney injury (AKI) imposes a significant medical burden. Due to the lack of effective drug transport vehicles, the administration of therapeutic agents for AKI cannot obtain the desired therapeutic effects. Kidney-targeted nanoparticles for renal delivery of drugs have shown promising potential as an emerging strategy for AKI therapy. However, these exogenous nanoparticles are rapidly cleared in the body and fail to achieve the expected renal targeting efficiency. Herein, we prepared the kidney targeting peptide-modified renal tubular epithelial cell membrane to coat zeolite imidazolate framework-8 nanoparticles for FGF21 delivery (KMZ@FGF21) for AKI treatment. KMZ@FGF21 could be efficiently internalized by renal cells and exhibited antioxidative, antiapoptotic and anti-inflammatory effects. A septic AKI murine model was established to assess the in vivo performance of KMZ@FGF21. The results showed that injected KMZ@FGF21 specifically accumulated in the injured kidney and exerted good renoprotective effects. This study provides an innovative thread for precise drug delivery in the treatment of various renal diseases.

(187) Routine Staining of Excised Vestibular Specimens with Hematoxylin and Eosin and Immunohistochemical Staining with CD117 and PGP9.5: Added Value to Patients

Abstract Introduction Neuroproliferative vestibulodynia (NPV) is associated with increased density of mast cells in the epithelial basement membrane and nerve endings in the sub-epithelial stroma. As cellular pathology cannot be visualized on physical examination, diagnosis is difficult, and often delayed. Patients with entrance dyspareunia are suspected to have NPV only after excluding other forms of vestibulodynia. Surgical specimens from complete vestibulectomy with vaginal advancement flap reconstruction are sent to pathology for routine staining with hematoxylin and eosin (H&amp;E). Since 2019 we have also ordered immunohistochemical staining with CD117 and PGP9. Objective We wished to compare the pathologic findings in excised vestibular specimen examined by routine H&amp;E staining versus by CD117 and PGP9.5, and to assess the significance to the patient of having both pathologic examinations performed. Methods Patients with suspected neuroproliferative vestibulodynia who underwent complete vestibulectomy with vaginal advancement flap reconstruction had both routine H&amp;E staining and CD117/PGP9.5 staining of the excised specimen. Patients were asked post-operatively if and how this information benefitted them. Results Fifty-six patients formed the study group. Based on H&amp;E staining, all patients were diagnosed with benign squamous vestibular epithelium with chronic inflammation. Gross examination showed tissue sizes varying from 5-11 cm long x 0.5 – 4 cm wide x 0.1 – 0.7 cm thick. Microscopic examination showed chronic inflammatory infiltrate at the superficial sub-epithelial stroma (Fig 1). This was composed of mature appearing lymphocytes in 87%, and a mixture of lymphocytes and plasma cells in 13%. Eosinophils were noted in 25% cases. P16 immunohistochemical staining, performed to rule out HPV when appropriate, was negative in all cases. Focal parakeratosis was noted in 7% cases. No ulcers, dysplastic or malignant cells were noted in any specimen. Based on CD117 and PGP9.5 staining, the diagnosis of NPV was established (Figs 2, 3). Positive results consistent with excess mast cells, greater than 8 cells per high power field x400, were noted in 100% of cases. All patients (100%) valued the pathologic information, especially regarding the positive CD117 and PGP9.5 staining as final confirmation of their NPV diagnosis, in particular after experiencing medical gaslighting. They consistently used the term “validating”, providing “tangible evidence” of a “physical problem wrong with me” when they had been told their “pain didn’t exist”. “Having that ‘knowledge’ was priceless. It's one thing to have a belief that I had neuroproliferative vestibulodynia. It's another thing to KNOW. To have that indisputable proof is one of the most gratifying and emotional moments in my healing journey. It shows that there was something wrong. It wasn't all in my head. I didn't need to ‘just relax’.” Conclusions The clinical diagnosis of NPV is based on excluding other forms of vestibulodynia. The pathologic diagnosis of NPV is not based on routine pathology staining but on immunohistochemical staining with CD117 and PGP9.5. If a patient is ignored, misdiagnosed, or invalidated by a provider, the patient loses self-confidence and faith in their own abilities. Providing pathologic evidence of the root cause of entrance dyspareunia plays a significant role in the healing process. Disclosure No

(136) Long-term Durability of Response to Complete Vestibulectomy Surgery for Neuroproliferative Vestibulodynia

Abstract Introduction In 2006, Goldstein et al published on the surgical outcomes of the largest-to-date patient series undergoing complete vestibulectomy with vaginal advancement flap reconstruction for neuroproliferative vestibulodynia (NPV). One hundred and thirty-four patients underwent surgery over a 5 year period, and 93% of the 104 patients who agreed to participate were satisfied/very satisfied with their surgical outcome. The mean time to follow up was 26 months, range 12 – 72 months. NPV is hypothesized to be, in part, a result of accumulation of excess mast cells in the epithelial basement membrane and sub-epithelial stroma that may release nerve growth factor promoting sub-epithelial nerve ending proliferation (Figure 1). The excess nerve endings are hypothesized to result in bothersome, persistent, and unrelenting provoked vestibular allodynia and hyperalgesia that are characteristic of NPV. Durability of post-operative outcomes past 72 months has yet to be addressed. It is still unknown if the pathologic findings of excess mast cells and nerve endings ever regenerate in the vestibule and cause recurrent allodynia, hyperalgesia and entrance dyspareunia. Objective We wished to assess, for the first time, the long-term surgical outcome durability data (&amp;gt;5 years) of those NPV patients who claimed successful resolution of their entrance dyspareunia following complete vestibulectomy surgery in the first 5 years post-operatively. Methods We routinely perform post-operative follow-up of surgical patients who undergo complete vestibulectomy surgery with vaginal advancement flap reconstruction for NPV. We identified those who claimed successful resolution of their entrance dyspareunia following complete vestibulectomy surgery in the first year post-operatively and thereafter for the next 4 years, and asked them if they continued to have resolution of their entrance dyspareunia. Results Fifty-seven patients had complete vestibulectomy with vaginal advancement flap reconstruction between 2007 and 2017 (&amp;gt; 5 years ago). Forty-one patients responded to the routine contact for long term follow-up. Thirty-four patients (78%) had resolution of pain in the first year post-operatively. Upon repeat interview, all 34 (100%) continued to note excellent durability of their surgical result, a mean of 9.3 years after their vestibulectomy (range 5-14 years). Conclusions Based on this small series, it appears that if a patient is a treatment responder following complete vestibulectomy, there is strong durability of the surgical result with no evidence of recurrent allodynia or hypoalgesia. These data suggest that in treatment responders, there is no evidence of re-accumulation of excess mast cells and nerve endings in the introitus. These data support that the pathology of NPV lies in the endodermal vestibule and that complete excision may be an important long-term treatment strategy in many NPV patients. Disclosure No