CD31 Immunohistochemical Staining Research Articles

Epstein-Barr Virus Lytic Transcripts Correlate with the Degree of Myocardial Inflammation in Heart Failure Patients

The Epstein-Barr virus (EBV) is frequently found in endomyocardial biopsies (EMBs) from patients with heart failure, but the detection of EBV-specific DNA has not been associated with progressive hemodynamic deterioration. In this paper, we investigate the use of targeted next-generation sequencing (NGS) to detect EBV transcripts and their correlation with myocardial inflammation in EBV-positive patients with heart failure with reduced ejection fraction (HFrEF). Forty-four HFrEF patients with positive EBV DNA detection and varying degrees of myocardial inflammation were selected. EBV-specific transcripts from EMBs were enriched using a custom hybridization capture-based workflow and, subsequently, sequenced by NGS. The short-read sequencing revealed the presence of EBV-specific transcripts in 17 patients, of which 11 had only latent EBV genes and 6 presented with lytic transcription. The immunohistochemical staining for CD3+ T lymphocytes showed a significant increase in the degree of myocardial inflammation in the presence of EBV lytic transcripts, suggesting a possible influence on the clinical course. These results imply the important role of EBV lytic transcripts in the pathogenesis of inflammatory heart disease and emphasize the applicability of targeted NGS in EMB diagnostics as a basis for specific treatment.

Application of Stem Cells Shows Antiinflammatory Effect in an Irradiated Random Pattern Flap Model

Background: In reconstructive surgery, local flaps might develop tissue necrosis or partial flap loss especially after previous irradiation, which may be necessary in many tumor entities. The application of stem cells seems promising to improve flap perfusion and might be a possible solution to optimize flap survival. Methods: Twenty rats received harvesting of bilateral random pattern fasciocutaneous flaps. The right flaps received 20 Gy ionizing radiation 4 weeks prior to the surgery, while the left flaps served as the non-irradiated control. After flap harvest, four different stem cell mixtures (5 × 106 ASC, ASC-HUVEC, MSC, MSC-HUVEC) were applied under both right and left flaps using 1 mL fibrin glue as the delivery vehicle. Flap size and its necrotic area were examined clinically. Two weeks after the surgery, HE staining and immunohistochemical staining for CD68 and ERG, as well as PCR analysis (Interleukin 6, HIF-1α and VEGF), were performed. Results: Application of ASCs, ASCs-HUVECs and MSCs resulted in a lower number of CD68-stained cells compared to the no cell group. The expression of Hif1α was higher in the ASC group compared to those in the MSC and previously treated no cell groups. Treatment with MSCs and MSCs-HUVECs prevented shrinking of the flaps in this series. Conclusion: Application of ASCs, MSCs and ASCs-HUVECs was shown to have an antiinflammatory effect. Treatment with MSCs and MSCs-HUVECs can prevent early shrinking of the flaps.

Solitary primary intraosseous xanthoma of the mandible in a 15-year-old boy: a case report

BackgroundA xanthoma is a rare bone condition consisting of a predominant collection of lipid-rich, foamy histiocytes. The central xanthoma of the jaws is a unique benign tumor.Case reportA 15-year-old Caucasian male has been presented to our department. He had radiological changes in the area of the left mandibular angle, with an area of diffuse osteolysis of 3.0 cm by 2.0 cm. Computed tomography reveals an area of diffuse osteolysis that starts from the distal root of the lower second molar and reaches the ascending process. A bone biopsy was performed, which revealed a benign proliferative process composed of histiocytic cells involving and infiltrating trabecular bone in a background of loose fibrous connective tissue devoid of any other significant inflammatory infiltrate. The size of the formation was 2.9 cm by 2.0 cm. Immunohistochemical staining for CD68 was strongly positive and negative for S-100 and CD1a. From routine blood tests, cholesterol, triglycerides, and blood sugar are within normal values, which excludes systemic metabolic disease. Subsequent to the surgical intervention, the patient underwent postoperative assessments at intervals of 14, 30, 60 days, and a year later, revealing the absence of any discernible complications during the aforementioned observation periods.ConclusionThe diagnosis of primary xanthoma of the mandible is rare and can often be confused with other histiocytic lesions. A differential diagnosis should be made with nonossifying fibroma and Langerhans cell histiocytosis, as in our case. In these cases, immunohistochemistry with CD 68, S-100, and CD1a, as well as blood parameters, are crucial for the diagnosis.

Time-restricted feeding does not improve nephropathy in humanized sickle cell disease mice

Sickle cell disease (SCD) is an autosomal recessive genetic disease caused by a point mutation in the hemoglobin gene. This substitution results in a “sickling” of red blood cells and subsequent vascular and organ damage. Current treatments, while increasing the lifespan of sickle cell patients, has resulted in a higher prevalence of chronic complications like SCD nephropathy. Our group has previously shown that time-restricted feeding (TRF) in chronic high fat fed mice reduces kidney damage. Given the increased kidney damage in SCD, we hypothesize that TRF in humanized SCD mice will attenuate sickle cell nephropathy. To test this hypothesis, we studied 18-22-week-old male SCD mice. Mice underwent 3 weeks of TRF, only having access to food during the active, dark period (Zeitgeber Time (ZT) 12 to 24). The control group had access to food ad libitum (ad-lib). At the end of the 3 weeks, at 21-25 week old ages, 12 hour urine was collected in metabolic cages as well as kidneys and plasma collection at ZT3 (inactive period). Excretion of urinary kidney damage markers (albumin and NGAL-1) showed no difference between TRF and ad-lib in both the active and inactive collection periods. Urinary excretion of endothelin-1 is known to be upregulated in SCD patients; however, TRF did not change this marker in SCD mice. TRF did not change plasma levels of cytokines or soluble adhesion molecules. Histological analysis of kidneys was conducted as an additional marker of kidney damage. We analyzed iron levels in the kidney with Prussian blue staining as well as fibrosis with Masson’s trichrome. TRF did not change kidney iron accumulation or kidney fibrosis. Immunohistochemical staining for CD3+ T cells did not show a difference in glomerular T cell infiltration in the TRF group compared to the ad-lib group. In conclusion, TRF does not appear to have an effect on renal inflammation or kidney function in SCD mice, but future studies need to be completed. R01 DK134562. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Immuno-histochemical evaluation of CD34 for OLP and OSMF

Immuno-histochemical evaluation of CD34 in oral lichen planus (OLP) and Oral Submucous Fibrosis (OSMF) is of interest to dentist.20 specimens of normal oral mucosa (buccal mucosa/gingiva tissue) from patients who had extractions performed as part of orthodontic treatment comprised Group I, the control group. Group II comprised 30 individuals with a diagnosis of oral lichen planus. 30 OSMF cases with diagnoses is Group III. These 80 specimens were all given consideration when choosing for CD34immuno-histochemical staining. The CD34 was greater in all categories of OLP and OSMF when compared to normal controls. Maximum CD34 expression was observed in erosive OLP (147.41ą17.60) followed by OSMF (116.01 ą11.72) and reticular OLP (105.01ą11.62). Data was statistically significant (p<0.001).Immunohistochemistry of CD34 evaluation is a potential diagnostic marker for OLP and OSMF.

Effects of applying human umbilical cord mesenchymal stem cell exosomes through different pathways to treat full-thickness skin defect wounds in mice

Objective: To investigate the effects of human umbilical cord mesenchymal stem cell (hUCMSC) exosomes in the treatment of full-thickness skin defect wounds in mice through local wound application, subcutaneous injection at the wound margin, and tail vein injection, and to explore the optimal administration route of hUCMSC exosomes for wound treatment. Methods: This study was an experimental study. hUCMSC exosomes were extracted from the discarded umbilical cord tissue of three normal delivery women aged 25-35 years in the Department of Obstetrics and Gynecology of Baogang Hospital of Inner Mongolia and successfully identified. Totally 120 male BALB/c mice aged 6-8 weeks were selected, and full-thickness skin defect wounds were prepared on the back of them. According to the random number table, the injured mice were divided into control group (without drug administration), local wound application group, wound margin subcutaneous injection group, and tail vein injection group (with 30 mice in each group). Mice in the latter three groups were given 0.2 mL phosphate buffer solution containing 200 μg hUCMSC exosomes by local wound application, subcutaneous injection at the wound margin, and tail vein injection, respectively. On post injury day (PID) 7, 14, and 21, the general condition of the wound was observed, and the wound healing rate was calculated; the wound tissue was collected, the pathological changes and collagen fibers were observed respectively by hematoxylin-eosin staining and Masson staining, the number of new microvessels was observed by CD31 immunohistochemical staining, and the content of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) was detected by enzyme-linked immunosorbent assay. The sample number was 10 in each group at each time point. Results: On PID 7, 14, and 21, the wounds of mice in the 4 groups all healed gradually, and the wound healing of the mice in wound margin subcutaneous injection group was the best; the wound healing rates of mice in the three administration groups were significantly higher than those in control group (P<0.05), the wound healing rates of mice in wound margin subcutaneous injection group and tail vein injection group were significantly higher than those in local wound application group (P<0.05), and the wound healing rates of mice in wound margin subcutaneous injection group were significantly higher than those in tail vein injection group (P<0.05). On PID 7, 14, and 21, the growth and epithelialization speed of the wound tissue of mice in the three administration groups were significantly accelerated, and the collagen fibers in the wounds of mice in the three administration groups were larger in number and more neatly arranged in comparison with the control group. On PID 7, 14, and 21, under every 200-fold visual field, the number of new microvessels in the wound tissue of mice in local wound application group was 24.1±2.5, 50.7±4.1, and 44.2±2.3, respectively, the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group was 32.2±2.9, 67.5±4.9, and 53.6±3.7, respectively, and the number of new microvessels in the wound tissue of mice in tail vein injection group was 27.8±2.4, 59.1±3.7, and 49.6±2.6, respectively, which was significantly more than 20.6±1.7, 46.7±3.4, and 40.9±2.8 in control group (P<0.05); the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group and tail vein injection group was significantly more than that in local wound application group (P<0.05); the number of new microvessels in the wound tissue of mice in wound margin subcutaneous injection group was significantly more than that in tail vein injection group (P<0.05). On PID 7, 14, and 21, the content of TNF-α and IL-6 in the wound tissue of mice in the three administration groups was significantly less than that in control group (P<0.05), the content of TNF-α and IL-6 in the wound tissue of mice in wound margin subcutaneous injection group and tail vein injection group was significantly less than that in local wound application group (P<0.05), and the content of TNF-α and IL-6 in the wound tissue of mice in wound margin subcutaneous injection group was significantly less than that in tail vein injection group (P<0.05). Conclusions: Local wound application, subcutaneous injection at the wound margin, and tail vein injection of hUCMSC exosomes can all promote the wound healing of full-thickness skin defects in mice through alleviating excessive inflammatory response and promoting angiogenesis. Among them, subcutaneous injection at the wound margin has a better therapeutic effect, indicating subcutaneous injection at the wound margin is the optimal administration route for hUCMSC exosomes in wound treatment.

“ludwigia sedioides” inspired substance-exchangeable spacer facilitating revascularization for skin repair

Adequate substance exchange is an important prerequisite for tissue repair, and how to efficiently establish mature vascular networks to enhance the graft-host substance exchange is a challenge. Herein, inspired by the special structure of the aquatic plant “Ludwigia sedioides”, ultra-thin microporous scaffolds (TMS) of bovine Achilles tendon derived collagen were fabricated as “floating leaves” via pyridine reaction, and cleverly combined with platelet-rich plasma (PRP) after the Schiff base and Michael additions reactions of polydopamine to construct a biomimetic “Ludwigia sedioides” spacer (PTMS@P), allowing blood vessels (as a “rhizome”) freely shuttling and substance exchange. In this spacer, growth factors were efficiently released to peak concentrations in the first 12 h, thereby facilitating rapid blood vessel growth. In the subsequent 96 h, these slow-released growth factors retained at 11.6 % to 45.8 % of the peak concentrations, sustainedly promoting vascular networks shuttling through the spacer, graft-host substance exchange and extracellular matrix synthesis in fibroblasts. In vitro, the spacer significantly fostered fibroblasts and endothelial cells’ proliferation, adhesion and migration, which were fundamental for revascularization and substance exchange. Immunohistochemical staining of CD31 and Ki67 in vivo demonstrated that abundant vascular networks shuttled through PTMS@P and extended from flap base to the farther flap body, promoting more robust substance exchange and cellular metabolism, which significantly enhanced the survival of ischemic flaps in nude mice. Altogether, this “Ludwigia sedioides” inspired spacer PTMS@P with strengthened revascularization and substance-exchange functionality holds promising applications for tissue repair and regeneration.

Proteomics study of primary and recurrent adamantinomatous craniopharyngiomas

BackgroundAdamantinomatous craniopharyngiomas (ACPs) are rare benign epithelial tumours with high recurrence and poor prognosis. Biological differences between recurrent and primary ACPs that may be associated with disease recurrence and treatment have yet to be evaluated at the proteomic level. In this study, we aimed to determine the proteomic profiles of paired recurrent and primary ACP, gain biological insight into ACP recurrence, and identify potential targets for ACP treatment.MethodPatients with ACP (n = 15) or Rathke’s cleft cyst (RCC; n = 7) who underwent surgery at Sanbo Brain Hospital, Capital Medical University, Beijing, China and received pathological confirmation of ACP or RCC were enrolled in this study. We conducted a proteomic analysis to investigate the characteristics of primary ACP, paired recurrent ACP, and RCC. Western blotting was used to validate our proteomic results and assess the expression of key tumour-associated proteins in recurrent and primary ACPs. Flow cytometry was performed to evaluate the exhaustion of tumour-infiltrating lymphocytes (TILs) in primary and recurrent ACP tissue samples. Immunohistochemical staining for CD3 and PD-L1 was conducted to determine differences in T-cell infiltration and the expression of immunosuppressive molecules between paired primary and recurrent ACP samples.ResultsThe bioinformatics analysis showed that proteins differentially expressed between recurrent and primary ACPs were significantly associated with extracellular matrix organisation and interleukin signalling. Cathepsin K, which was upregulated in recurrent ACP compared with that in primary ACP, may play a role in ACP recurrence. High infiltration of T cells and exhaustion of TILs were revealed by the flow cytometry analysis of ACP.ConclusionsThis study provides a preliminary description of the proteomic differences between primary ACP, recurrent ACP, and RCC. Our findings serve as a resource for craniopharyngioma researchers and may ultimately expand existing knowledge of recurrent ACP and benefit clinical practice.

The water-soluble TF3 component from Eupolyphaga sinensis Walker promotes tibial fracture healing in rats by promoting osteoblast proliferation and angiogenesis

ObjectiveTo determine the active components of Eupolyphaga sinensis Walker (Tu Bie Chong) and explore the mechanisms underlying its fracture-healing ability. MethodsA modified Einhorn method was used to develop a rat tibial fracture model. Progression of bone healing was assessed using radiological methods. Safranin O/fast green and CD31 immunohistochemical staining were performed to evaluate the growth of bone cells and angiogenesis at the fracture site. Methylthiazoletetrazolium blue and wound healing assays were used to analyze cell viability and migration. The Transwell assay was used to explore the invasion capacity of the cells. Tubule formation assays were used to assess the angiogenesis capacity of human vascular endothelial cells (HUVECs). qRT-PCR was used to evaluate the changes in gene transcription levels. ResultsTu Bie Chong fraction 3 (TF3) significantly shortened the fracture healing time in model rats. X-ray results showed that on day 14, fracture healing in the TF3 treatment group was significantly better than that in the control group (P = .0086). Tissue staining showed that cartilage growth and the number of H-shaped blood vessels at the fracture site of the TF3 treatment group were better than those of the control group. In vitro, TF3 significantly promoted the proliferation and wound healing of MC3T3-E1s and HUVECs (all P < .01). Transwell assays showed that TF3 promoted the migration of HUVECs, but inhibited the migration of MC3T3-E1 cells. Tubule formation experiments confirmed that TF3 markedly promoted the ability of vascular endothelial cells to form microtubules. Gene expression analysis revealed that TF3 significantly promoted the expression of VEGFA, SPOCD1, NGF, and NGFR in HUVECs. In MC3T3-E1 cells, the transcript levels of RUNX2 and COL2A1 were significantly elevated following TF3 treatment. ConclusionTF3 promotes fracture healing by promoting bone regeneration associated with the RUNX2 pathway and angiogenesis associated with the VEGFA pathway.

Abstract 7667: Neutrophil to lymphocyte ratio (NLR) as an indicator of tumor immune status in non-small cell lung cancer

Abstract Background: Neutrophil to lymphocyte ratio (NLR) has been reported as a prognostic biomarker in non-small cell lung cancer (NSCLC), while its biological rationale has yet to be clarified. In cancer patients, inflammation and immune status plays crucial roles in tumor growth and the prognosis. While several studies have explored the relationship between NLR and inflammation, there are few reports on the relationship between NLR and immune status in the tumor microenvironment of cancer patient so far. The purpose of this study is to explore the potential utility of NLR as a surrogate biomarker for assessing the immune response to tumors. Methods: The medical records of NSCLC patients who underwent surgery at our institution in 2012 were retrospectively reviewed. The patients’ clinical information including age, sex, performance status, smoking history, histology, clinical stage (7th), and NLR, was obtained. NLR was calculated based on blood tests conducted within a month before surgery. Tumor immune status was evaluated by Immunohistochemical staining for CD3, CD8, and FOXP3 in their surgical specimens. The relationship between NLR, clinicopathology, and tumor immune status was examined. The survival curves were estimated using the Kaplan-Meier method and compared using the log-rank test. Blood cell counts and ratios between different groups were compared using nonpaired t test or Mann-Whitney U test, with a significance threshold set at p = 0.05. Results: A total of 120 patients were included. The 5-year overall survival rate (OS) in all 120 patients was 63.4%. The 5-year OS of the patients with lower NLR was significantly better than those with high NLR (low &amp;lt; 2.2 vs high ≥ 2.2; 70.1% vs 56.8%, p = 0.042). The 5-year OS of the patients with high density of CD3+ TILs was significantly better than that in patients with low density of CD3+ TILs (high ≥ 242 vs low &amp;lt; 242; 70% vs 56.8%, p = 0.019). The 5-year OS of the patients with high density of CD8+ TILs was significantly better than that in patients with low density of CD8+ TILs (high ≥ 112 vs low &amp;lt; 112; 72.4% vs 54.1%, p = 0.011). Conversely, the 5-year OS of the patients with low density of FOXP3+ TILs was significantly better than that in patients with high density of FOXP3+ TILs (high ≥ 7.5 vs low &amp;lt; 7.5; 46.2.% vs 79.6%, p = 0.0006). The number of CD3+ TILs had negative correlation with NLR (p = 0.005), while the number of CD8+ TILs and FOXP3+ TILs didn’t exhibit an association with NLR. Conclusion: Our results suggest that NLR might potentially reflect immune status in the tumor microenvironment, explaining its impact on the prognosis of NSCLC patients. While further research is needed, these findings may offer valuable insights for guiding treatment options. Citation Format: Ryo Yoshichika, Kazuma Iwata, Ken Suzawa, Yin Min Thu, Daisuke Mizuno, Tomoaki Higashihara, Naohiro Hayashi, Atsushi Matsuoka, Mao Yoshikawa, Fumiaki Mukohara, Kazuhiko Shien, Hiromasa Yamamoto, Shinichi Toyooka. Neutrophil to lymphocyte ratio (NLR) as an indicator of tumor immune status in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7667.

Stepwise degradable PGA-SF core-shell electrospinning scaffold with superior tenacity in wetting regime for promoting bone regeneration

Regenerating bone in the oral and maxillofacial region is clinically challenging due to the complicated osteogenic environment and the limitation of existing bone graft materials. Constructing bone graft materials with controlled degradation and stable mechanical properties in a physiological environment is of utmost importance. In this study, we used silk fibroin (SF) and polyglycolic acid (PGA) to fabricate a coaxial PGA-SF fibrous scaffold (PGA-SF-FS) to meet demands for bone grafts. The SF shell exerted excellent osteogenic activity while protecting PGA from rapid degradation and the PGA core equipped scaffold with excellent tenacity. The experiments related to biocompatibility and osteogenesis (e.g., cell attachment, proliferation, differentiation, and mineralization) demonstrated the superior ability of PGA-SF-FS to improve cell growth and osteogenic differentiation. Furthermore, in vivo testing using Sprague-Dawley rat cranial defect model showed that PGA-SF-FS accelerates bone regeneration as the implantation time increases, and its stepwise degradation helps to match the remodeling kinetics of the host bone tissue. Besides, immunohistochemical staining of CD31 and Col-1 confirmed the ability of PGA-SF-FS to enhance revascularization and osteogenesis response. Our results suggest that PGA-SF-FS fully utilizing the advantages of both components, exhibites stepwise degradation and superior tenacity in wetting regime, making it a promising candidate in the treatment of bone defects.

HMGA1 regulates the mitochondrial apoptosis pathway in sepsis-induced cardiomyopathy.

High mobility group protein AT-hook 1 (HMGA1), an architectural transcription factor, has previously been reportedto play an essential role in architectural remodeling processes. However, its effects on cardiovascular diseases, particularly sepsis-induced cardiomyopathy, have remained unclear. The study aimed to investigate the role of HMGA1 in lipopolysaccharide-induced cardiomyopathy. Mice subjected to lipopolysaccharide for 12 h resulted in cardiac dysfunction. We used an adeno-associated virus 9 delivery system to achieve cardiac-specific expression of the HMGA1 gene in the mice. H9c2 cardiomyocytes were infected with Ad-HMGA1 to overexpress HMGA1 or transfected with si-HMGA1 to knock down HMGA1. Echocardiography was applied to measure cardiac function. RT-PCR was used to detect the transcriptional level of inflammatory cytokines. CD45 and CD68 immunohistochemical staining were used to detect inflammatory cell infiltration and TUNEL staining to evaluate the cardiomyocyte apoptosis, MitoSox was used to detect mitochondrial reactive oxygen species, JC-1 was used todetect Mitochondrial membrane potential. Our findings revealed that the overexpression of HMGA1 exacerbated myocardial inflammation and apoptosis in response to lipopolysaccharide treatment. Additionally, we also observed that H9c2 cardiomyocytes with HMGA1 overexpression exhibited enhanced inflammation and apoptosis upon stimulation with lipopolysaccharide for 12 h. Conversely, HMGA1 knockdown in H9c2 cardiomyocytes attenuated lipopolysaccharide-induced cardiomyocyte inflammation and apoptosis. Further investigations into the molecular mechanisms underlying these effects showed that HMGA1 promoted lipopolysaccharide-induced mitochondrial-dependent cardiomyocyte apoptosis. The study reveals that HMGA1 worsens myocardial inflammation and apoptosis in response to lipopolysaccharide treatment. Mechanically, HMGA1 exerts its effects by regulating the mitochondria-dependent apoptotic pathway.

LncRNA XIST Interacts with Regulatory T Cells within the Tumor Microenvironment in Chronic Hepatitis B-Associated Hepatocellular Carcinoma.

Alterations in the expression of several long non-coding RNAs (lncRNAs) have been shown in chronic hepatitis B-associated hepatocellular carcinoma (CHB-HCC). Here, we aimed to investigate the association between the expression of inflammation-associated lncRNA X-inactive specific transcript (XIST) and the type of inflammatory cells within the tumor microenvironment. Twenty-one consecutive cirrhotic patients with CHB-HCC were included. XIST expression levels were investigated on formalin-fixed paraffin-embedded (FFPE) tumoral and peritumoral tissue samples by real-time polymerase chain reaction (RT-PCR). Immunohistochemical staining for CD3, CD4, CD8, CD25, CD163, CTLA4, and PD-1 were performed. The findings were statistically analyzed. Of the 21 cases, 11 (52.4%) had tumoral and 10 (47.6%) had peritumoral XIST expression. No significant association was found between the degree of inflammation and XIST expression. The number of intratumoral CD3, CD4, CD8 and CD20 positive cells was higher in XIST-expressing tumors, albeit without statistical significance. Tumoral and peritumoral XIST expression tended to be more common in patients with tumoral and peritumoral CD4high inflammation. The number of intratumoral CD25 positive cells was significantly higher in XIST-expressing tumors (p=0.01). Tumoral XIST expression was significantly more common in intratumoral CD25high cases (p=0.04). Peritumoral XIST expression was also more common among patients with CD25high peritumoral inflammation, albeit without statistical significance (p=0.19). lncRNA XIST is expressed in CHB-HCC and its expression is significantly associated with the inflammatory tumor microenvironment, particularly with the presence and number of CD25 (+) regulatory T cells. In vitro studies are needed to explore the detailed mechanism.

A Review for Dermatofibrosarcoma Protuberans

Dermatofibrosarcoma protuberans (DFSP) stands as the most prevalent cutaneous sarcoma, marked by slow growth and a protuberant papulonodular appearance. Although rare, its impact is significant, necessitating a comprehensive understanding of its characteristics and optimal management. DFSP typically affects adults, though age variability exists, presenting diagnostic challenges. Histological evaluation, often utilizing CD34 immunohistochemical staining, is crucial for accurate diagnosis. The primary treatment modality remains complete surgical excision, emphasizing the importance of generous margins. Imatinib emerges as a valuable therapeutic option for unresectable or metastatic cases. Complementary approaches such as adjuvant radiation therapy and Mohs surgery enhance the treatment armamentarium, allowing for a tailored, multidisciplinary strategy. Regular follow-up is essential for monitoring and addressing potential complications. While local recurrence is a concern, DFSP generally carries a favorable prognosis, especially with early detection and appropriate intervention. Ongoing research continues to refine treatment paradigms, promising improved outcomes for individuals affected by this distinctive cutaneous sarcoma.

Bifunctional tissue-engineered composite construct for bone regeneration: The role of copper and fibrin.

Bifunctional tissue engineering constructs promoting osteogenesis and angiogenesis are essential for bone regeneration. Metal ion-incorporated scaffolds and fibrin encapsulation attract much attention due to low cost, nontoxicity, and tunable control over ion and growth factor release. Herein, we investigated the effect of Cu.nHA/Cs/Gel scaffold and fibrin encapsulation on osteogenic and angiogenic differentiation of Wharton's jelly mesenchymal stem cells (WJMSCs) in vitro and in vivo. Cu-laden scaffolds were synthesized using salt leaching/freeze drying and were characterized using standard techniques. WJMSCs were isolated from the human umbilical cord and characterized. WJMSCs with or without encapsulating in fibrin were seeded onto scaffolds, followed by differentiating into the osteogenic lineage for 7 and 21 days. Osteogenic and angiogenic differentiation were evaluated using real-time polymerase chain reaction, western blot, and Alizarin red staining. Then, scaffolds were implanted into critical-sized calvarial bone defects in rats and histological assessments were performed using hematoxylin/eosin, Masson's trichrome, and CD31 immunohistochemical staining at 4 and 12 weeks. The scaffolds had good physicochemical and biological characteristics suitable for cell attachment and growth. Cu and fibrin increased the expression of ALP, RUNX2, OCN, COLI, VEGF, and HIF1α in differentiated WJMSCs. Implanted scaffolds were also biocompatible and were integrated well with the host tissue. Increased collagen condensation, mineralization, and blood vessel formation were observed in Cu-laden scaffolds. The fibrin-encapsulated groups showed the highest collagen and cell densities, immune cell infiltration, and bone trabeculae. CD31-positive cell population increased with fibrin encapsulation and seeding onto Cu-laden scaffolds. Adding Cu to scaffolds and encapsulating cells in fibrin are promising methods that guide osteogenesis and angiogenesis cellular signaling, leading to better bone regeneration.

Polycondensed Peptide-Based Polymers for Targeted Delivery of Anti-Angiogenic siRNA to Treat Endometriosis.

Endometriosis (EM) is a prevalent gynecological disease characterized by the abnormal growth of tissue similar to the endometrium outside of the uterus. This condition is accompanied by the development of new blood vessels in endometriotic lesions. While surgical intervention is effective in removing endometriotic lesions, some patients require multiple surgeries. Therefore, finding non-surgical treatments for EM is of great interest. One of the promising approaches is anti-angiogenic therapy using siRNA-therapeutics to target the expression of the VEGFA gene. Peptide-based polymers have shown promise as siRNA delivery systems due to their biocompatibility and ease of modification. We conducted a study to evaluate the effectiveness of the R6p-cRGD peptide carrier as a non-viral vehicle for delivering siRNA to endothelial cells in vitro and endometrial implants in vivo. We investigated the physicochemical properties of the siRNA-complexes, assessed cellular toxicity, and examined the efficiency of GFP and VEGFA genes silencing. Furthermore, we tested the anti-angiogenic effects of these complexes in cellular and animal models. The transfection with siRNA complexes led to a significant increase in VEGFA gene knockdown efficiency and a decrease in the migration of endothelial cells. For the animal model, we induced endometriosis in rats by transplanting endometrial tissue subcutaneously. We evaluated the efficiency of anti-angiogenic therapy for EM in vivo using anti-VEGF siRNA/R6p-RGD complexes. During this assessment, we measured the volume of the implants, analyzed VEGFA gene expression, and conducted CD34 immunohistochemical staining. The results showed a significant decrease in the growth of endometriotic implants and in VEGFA gene expression. Overall, our findings demonstrate the potential of the R6p-cRGD peptide carrier as a delivery system for anti-angiogenic therapy of EM.

Presence of crown-like structures in breast adipose tissue; differences between healthy controls, BRCA1/2 gene mutation carriers and breast cancer patients.

Crown-like structures (CLS) in breast adipose tissue are associated with inflammation and a potential factor in breast cancer behaviour. Whether this effect varies between breast cancer subtypes and is influenced by BMI and BRCA mutation status is presently unknown. Therefore, we compared CLS presence between adipose tissue of healthy controls, BRCA1/2 gene mutation carriers and breast cancer patients, and assessed the relation of CLS with clinical outcome in breast cancer patients. Immunohistochemical staining for CD68 was performed on breast adipose tissue sections of 48 healthy controls, 78 BRCA1/2 gene mutation carriers and 259 breast cancer patients. CLS presence and index (CLS/cm2) were correlated with BMI, BRCA status, tumour presence, intrinsic tumour subtype and tumour characteristics. Associations with clinical outcome were assessed. CLS were more often present in breast cancer patients compared to BRCA carriers and healthy controls. CLS presence was associated with the presence of breast cancer and high BMI. CLS were more often present in Luminal-B-like tumours compared to the other subtypes. No correlations between CLS and BRCA status or age was found. In TNBC, CLS were related to lymphovascular invasion. No association with survival was found. In conclusion, CLS were more frequently present in breast adipose tissue of breast cancer patients compared to BRCA1/2 gene mutation carriers and healthy controls. Furthermore, our study provides evidence of the association between obesity and presence of CLS. The prognostic significance and impact on clinical outcome of differences in CLS numbers should be further assessed in prospective studies.

Real-World Data of Outcome of Patients with Newly Diagnosed Bronchial-Associated Lymphoid Tissue Lymphoma: A Multi-Center Retrospective Analysis with Long-Term Survivors

Introduction Extranodal marginal-zone lymphoma (MZL) of mucosa-associated lymphoid tissue (MALT) is an indolent type of non-Hodgkin's lymphoma. Although the lung is the second most common site, it accounts for less than 1% of all lymphomas. Extranodal MZL of MALT type of the lungs originates from bronchial-associated lymphoid tissue (BALT) and is clinically referred to as “BALT lymphoma” or “BALToma”. The indolent nature of BALTomas results in none or mild symptoms and their slow growth pattern allows for various treatment approaches. Depending on the case, active surveillance may be considered, while others may benefit from surgery, radiotherapy or chemotherapy. Despite some existing reviews on this lymphoma, comprehensive characterization of optimal management and prognostic factors remains inadequately defined. Limited case reports and small-scale retrospective data analyses have been conducted, but the number of patients included in these studies was highly restricted. In this study, we conducted a retrospective analysis of clinical data from 67 BALToma patients in South Korea. Methods 67 patients were included in our analysis, all of whom were diagnosed with BALToma between September 1996 and January 2023 at three institutions in South Korea. The criteria for case inclusion was histologically confirmed diagnosis of extranodal MZL of pulmonary origin in accordance with WHO classifications, and availability of clinical data. Extranodal MZL was diagnosed based on WHO classifications, along with the findings from immunohistochemical staining for CD20 and CD3. If needed, CD5, CD10, CD23, cyclin D1, BCL6, and Ki-67 was also stained. Gene rearrangement studies targeting the IgH gene were conducted using PCR analysis. Patients were treated according to the stage and location of the disease. The patients were analyzed into two groups; observation or localized treatment and systemic chemotherapy. Localized treatment included surgery or radiotherapy. Systemic chemotherapy regimens used were mainly (R)-CVP and (R)-CHOP. Baseline characteristics was analyzed with Fisher's exact test. Overall survival (OS) and Progression Free Survival (PFS) were estimated using the Kaplan-Meier method. PFS was calculated from the date of diagnosis to the date on which progression was noted, the date of the last follow-up, or the date of death. OS was measured from the date of diagnosis to the date of death or the last follow-up visit. Survival rates were compared for statistical differences via Generalized Wilcoxon analyses. Prognostic factor analysis and hazard ratio was calculated using univariate and multivariate Cox Regression method. All statistical tests were two-sided, and p &amp;lt; 0.05 was considered statistically significant. All data was analyzed using SPSS Statistics for Windows, Version 27.0. Results The clinical characteristics of patients according to treatment modality was analyzed (Table 1). Gender, age, performance status was not significantly different between each group. Notably, involvement of single or multiple lobe (p=0.045), involvement of bilateral lung (p&amp;lt;0.001), nodal involvement (p=0.033), and involvement of extrapulmonary organ (p=0.010) was significantly different between each group. The PFS and OS according to treatment method are provided in Figure 1 and 2. 10-year PFS for observation or localized treatment was 78.7% whereas chemotherapy was 56.9% (p=0.044). 10-year OS for observation or localized treatment was 100%, whereas chemotherapy was 71.7% (p=0.016). The prognostic factors for progression of disease were significant only for the involvement of extrapulmonary organ in both univariate and multivariate (hazard ratio 3.615, p=0.023) analysis (Table 2). Prognostic factor analysis for overall survival did not yield any significant result. Conclusion This study concludes that the most important prognostic factor for the progression of disease is the involvement of extrapulmonary organ. This study implies that the feasibility of simply monitoring patients whose disease is confined exclusively to the lung is a viable strategy. However, in the case of extrapulmonary involvement, the implementation of systemic chemotherapy ought to be deliberated. Given the heightened potential for disease progression, this underscores the necessity for meticulous consideration during the development of therapeutic plans.

The Immunoarchitecture of Human Extraocular Muscles.

The purpose of this study was to describe the immunoarchitecture of normal extraocular muscles (EOMs) in terms of presence, distribution, and organization of various immune cells. We performed unilateral orbital exenterations in six fresh human cadavers from elderly patients, followed by dissection of the medial, lateral, superior and inferior rectus, superior and inferior oblique, and superior palpebral levator muscle in their entirety. We further cross sectioned each EOM in an anterior, central, and posterior third. After immunohistochemical staining for CD3, CD8, CD20, CD138, CD68, and podoplanin, quantitative analysis was performed. We found all EOMs (rectus, oblique, and levator muscles) to harbor both T- and B-lymphocytes, with a B-lymphocyte dominance and an absence of plasma cells. The highest prevalence of immune cells was seen in the muscle bellies, with, on average, 488 ± 63 CD3+ T-lymphocytes and 44 ± 110 CD20+ B-lymphocytes per mm2, and significant differences from the anterior (T-lymphocytes) and posterior (T- and B-lymphocytes) thirds. T- and B-lymphocytes were primarily organized in hotspots in the vicinity of blood vessels. In addition, a small resident population of macrophages scattered throughout the specimens was detected. No lymphatic vessels were found in any of the EOMs. These findings can serve as a reference dataset in the assessment of EOM biopsies in the diagnostic process of inflammatory orbital and systemic disorders. Moreover, from a regenerative perspective, our results highlight the importance of taking into account the presence of a resident immune cell population when studying the host immune response on transplanted tissues or engineered constructs.

Kaposiform hemangioendothelioma presented with raynaud phenomenon: a case report

BackgroundKaposiform hemangioendothelioma (KHE) is a rare vascular neoplasm affecting infants or young children. KHE includes a spectrum of lesions, ranging from small and superficial tumors to large and invasive lesions with Kasabach-Merritt phenomenon (KMP). Currently, no published studies have reported a KHE presenting as thrombocytopenia and Raynaud phenomenon.Case presentationA 2-year-old boy with right hand swelling and thrombocytopenia was admitted to our hospital. His right hand turned swelling and red, even occasionally cyanotic. This condition became worse in response to cool environments and improved with warming, and platelet counts were between 50 ~ 80 × 10^9/L. Physical examination on admission revealed the swelling and frostbite-like rash of the right-hand fingers, and the skin temperature of the right hand was lower than the left. On day 3 of admission, chest CT results showed an irregular mass on the right side of the spine. The puncture biopsy demonstrated positive CD31, D2-40, and FLI1 immunohistochemical staining, but negative GLUT1 staining, confirming the diagnosis of KHE. Furthermore, endothelin-1 (ET1) expression levels significantly increased, and eNOS and A20 expression levels significantly decreased comparing with control patients. The patient received methylprednisolone and sirolimus treatments, and his condition gradually improved during the follow-up.ConclusionsWe reported the first case of KHE presenting with thrombocytopenia and Raynaud phenomenon. The development of Raynaud phenomenon could be associated with increased ET-1 and reduced eNOS and A20 expressions. Careful differential diagnosis of hidden KHE should be considered in children with thrombocytopenia and Raynaud phenomenon.