Progress in Microdermal Grafting for Color Regeneration of White Scars.

BACKGROUND: Microdermal grafting with knife-cut, partially de-epithelialized skin can regenerate color in white (hypopigmented) scars. However, the scalp has more melanocytes, and dermabrasion can preserve more melanocytes than knife cutting during partial de-epithelialization. OBJECTIVES: The aim of this study was to evaluate the color regeneration results and complications of various microdermal grafting procedures for white scar color regeneration. METHODS: Two refinements to an existing microdermal grafting technique for treating white scars were described: dermabrasion, rather than knife cutting, was used to partially de-epithelialize skin, and melanocyte donor sites were harvested from the scalp, rather than from skin. A review was performed of 65 cases in which various combinations of these refinements were used to treat scars on the face and forearms. RESULTS: Sixty-five patients (36 forearms; 29 faces) were treated, 40 receiving 1 session, 23 receiving 2 sessions, and 2 receiving 3 sessions of treatment. The follow-up was 6.5 months (range, 4-16 months). The use of both technique refinements produced approximately 15% better color generation than the original procedure after 1 session of treatment and approximately 20% better than the original procedure after 2 sessions. Histologic immunostaining showed that the dermabrasion method preserved more melanocytes around the epidermal-dermal region, and that the scalp has richer melanocytes than skin. The complication rate was reduced. CONCLUSIONS: The use of the scalp as the donor site and partial de-epithelialization by dermabrasion can be safely incorporated into a previously developed microdermal grafting procedure for better color regeneration of white scars.

Characterization of the pleural microenvironment niche and cancer transition using single-cell RNA sequencing in EGFR-mutated lung cancer.

Background: Lung cancer is associated with a high mortality rate and often complicated with malignant pleural effusion (MPE), which has a very poor clinical outcome with a short life expectancy. However, our understanding of cell-specific mechanisms underlying the pathobiology of pleural metastasis remains incomplete. Methods: We analyzed single-cell transcriptomes of cells in pleural effusion collected from patients with lung cancer and congestive heart failure (as a control), respectively. Soluble and complement factors were measured using a multiplex cytokine bead assay. The role of ferroptosis was evaluated by GPX4 small interfering RNA (siRNA) transfection and overexpression. Results: We found that the mesothelial-mesenchymal transition (MesoMT) of the pleural mesothelial cells contributed to pleural metastasis, which was validated by lung cancer/mesothelial cell co-culture experiments. The ferroptosis resistance that protected cancer from death which was secondary to extracellular matrix detachment was critical for pleural metastasis. We found a universal presence of immune-suppressive lipid-associated tumor-associated macrophages (LA-TAMs) with complement cascade alteration in the MPE of the lung cancer patients. Specifically, upregulated complement factors were also found in the MPE, and C5 was associated with poor overall survival in the lung cancer patients with epidermal growth factor receptor mutation. Plasmacytoid dendritic cells (pDCs) exhibited a dysfunctional phenotype and pro-tumorigenic feature in the primary cancer. High expression of the gene set extracted from pDCs was associated with a poor prognosis in the lung cancer patients. Receptor-ligand interaction analysis revealed that the pleural metastatic niche was aggravated by cross-talk between mesothelial cells-cancer cells/immune cells via TNC and ICAM1. Conclusions: Taken together, our results highlight cell-specific mechanisms involved in the pathobiological development of pleural metastasis in lung cancer. These results provide a large-scale and high-dimensional characterization of the pleural microenvironment and offer a useful resource for the future development of therapeutic drugs in lung cancer.

TRPM1 promotes tumor progression in acral melanoma by activating the Ca2+/CaMKIIδ/AKT pathway.

INTRODUCTION: Acral melanoma is a predominant and aggressive subtype of melanoma in non-Caucasian populations. There is a lack of genotype-driven therapies for over 50% of patients. TRPM1 (transient receptor potential melastatin 1), a nonspecific cation channel, is mainly expressed in retinal bipolar neurons and skin. Nonetheless, the function of TRPM1 in melanoma progression is poorly understood. OBJECTIVES: We investigated the association between TRPM1 and acral melanoma progression and revealed the molecular mechanisms by which TRPM1 promotes tumor progression and malignancy. METHODS: TRPM1 expression and CaMKII phosphorylation in tumor specimens were tested by immunohistochemistry analysis and scored by two independent investigators. The functions of TRPM1 and CaMKII were assessed using loss-of-function and gain-of-function approaches and examined by western blotting, colony formation, cell migration and invasion, and xenograft tumor growth assays. The effects of a CaMKII inhibitor, KN93, were evaluated using both in vitro cell and in vivo xenograft mouse models. RESULTS: We revealed that TRPM1 protein expression was positively associated with tumor progression and shorter survival in patients with acral melanoma. TRPM1 promoted AKT activation and the colony formation, cell mobility, and xenograft tumor growth of melanoma cells. TRPM1 elevated cytosolic Ca2+ levels and activated CaMKIIδ (Ca2+/calmodulin-dependent protein kinase IIδ) to promote the CaMKIIδ/AKT interaction and AKT activation. The functions of TRPM1 in melanoma cells were suppressed by a CaMKII inhibitor, KN93. Significant upregulation of phospho-CaMKII levels in acral melanomas was related to increased expression of TRPM1. An acral melanoma cell line with high expression of TRPM1, CA11, was isolated from a patient to show the anti-tumor activity of KN93 in vitro and in vivo. CONCLUSIONS: TRPM1 promotes tumor progression and malignancy in acral melanoma by activating the Ca2+/CaMKIIδ/AKT pathway. CaMKII inhibition may be a potential therapeutic strategy for treating acral melanomas with high expression of TRPM1.

Utility of the capsule-based technique for cell block preparation - in body fluids and Liqui-PREP™ specimens.

OBJECTIVE: The aim of this study was to find a better technique for the cell block preparation. STUDY DESIGN: We developed a novel capsule-based technique and applied it to different cell block preparation materials including plasma thromboplastin (PT) clot and agarose gel-embedding medium, in order to concentrate the cells in a limited field, then compared the result with the widely used, modified alcohol fixative preparation technique based on specimen adequacy and diagnostic accuracy. Twenty-eight nongynecologic body fluids and 41 gynecologic Liqui-PREP™ (LGM International Inc., Fort Lauderdale, Fla., USA) cytology specimens were collected for cell block preparation. We performed routine hematoxylin and eosin staining on the cell block sections, which were scored according to four specimen adequacy parameters: background, cellularity, nuclear preservation and cytoplasmic preservation. The differences between the diagnostic results of cell block slide and conventional cytology smear were also presented. RESULTS: The capsule-based PT clot technique in nongynecologic body fluids provided more cellularity in the cell block sections and reduced atypical diagnosis when compared to conventional smears. CONCLUSION: We suggest the capsule-based PT clot technique is a better technique for nongynecologic body fluids in the preparation of a satisfactory cell block.

Low Galectin-3 Expression Level in Primary Tumors Is Associated with Metastasis in T1 Lung Adenocarcinoma.

BACKGROUND AND OBJECTIVE: Although nodal and distant metastasis is rare in T1 lung adenocarcinoma, it is related to poor clinical prognosis. Association between galectin-3 (Gal-3) expression level, and clinical outcome of T1 lung adenocarcinoma has not been clarified. METHODS: From January 2009 to December 2014, 74 patients with surgically resected T1 lung adenocarcinoma were enrolled in this retrospective cohort study. Patient outcomes were followed up until December 2019. Gal-3 expression level in primary tumors was assessed immunohistochemically and evaluated based on the staining intensity and percentage. Patient characteristics and correlation between Gal-3 expression level and clinical outcomes were reviewed. RESULTS: Low Gal-3 expression was associated with increased metastatic events (p = 0.03), especially distant metastasis (p = 0.007), and mortality rate (p = 0.04). Kaplan-Meier analysis revealed that high Gal-3 expression level was associated with favorable recurrence-free survival in T1 lung adenocarcinoma (log-rank p = 0.048) and T1a (≤ 2 cm, American Joint Committee on Cancer (AJCC) 7th edition) lung adenocarcinoma (log-rank p = 0.043). Gal-3 expression along with tumor size showed a larger area under curve (AUC) than tumor size alone for predicting metastatic events (AUC = 0.747 vs. 0.681) and recurrence (AUC = 0.813 vs. 0.766) in T1a lung adenocarcinoma in the receiver-operating characteristic curve. CONCLUSION: Low Gal-3 expression level in primary tumors was remarkably associated with increased metastatic events and reduced recurrence-free survival in T1 lung adenocarcinoma. We suggest that Gal-3 expression level in addition to tumor size may potentially be stronger than tumor size alone in predicting metastasis in T1a lung adenocarcinoma patients.

Diagnostic value of computerized nuclear morphometry for the prediction of malignancy in liver fine needle aspiration cytology.

OBJECTIVE: To evaluate the diagnostic value of nuclear parameters using computerized morphometry in the differential diagnosis of well-differentiated hepatocellular carcinoma (WD-HCC) and regenerative atypia in liver fine needle aspiration cytology (FNAC). STUDY DESIGN: All 78 cases, including 30 negative, 30 positive and 18 suspicious liver FNA cytology smears were examined with regard to nuclear area, nuclear perimeter, largest to smallest diameter ratio of the nuclei (L/S ratio) and coefficient of variation of the nuclear area (NACV). The diagnostic value was assessed using receiver operating characteristics (ROC) curves. Cutoff points were determined by the area under the ROC curve (AUC), which provides the highest combined sensitivity and specificity. RESULTS: The nuclear area and perimeter in liver FNA were more helpful than L/S ratio and NACV in discriminating between hepatocytes with regenerative atypia and WD-HCC. Cutoff points of 56.67 for nuclear area and 27.16 for perimeter were suggested to help shift the suspicious diagnoses to WD-HCC. CONCLUSION: When faced with difficult diagnoses on liver FNA, we suggest application of computerized morphometry may be a helpful ancillary diagnostic tool in the further classification of such cases.

Cecal lipoma with subclinical appendicitis: A case report.

BACKGROUND: Colonic lipomas are rare, slow-growing benign tumors. Colonic lipomas are generally asymptomatic and are found incidentally. Although cases of cecal lipoma have been sporadically reported in the literature, the disease has not been systematically reviewed. CASE SUMMARY: We present a 44-year-old man who underwent a routine physical check-up during which colonoscopic examination revealed an asymptomatic 1.5-cm cecal mass at the appendiceal orifice. Laparoscopic exploration was performed that also demonstrated a congested and erythematous appendix. En bloc resection of both the cecum and vermiform appendix was performed because of the suspicion of malignancy. Histopathological examination revealed a cecal lipoma composed of mature adipose tissue, and the appendix showed subclinical inflammation. Our procedures and findings were discussed, along with relevant English literature that was retrieved from the PubMed database from 2000 to 2017. Twenty-six cases, including ours, were reported. Consistent with the findings of the literature, it is difficult to obtain a definitive diagnosis by colonoscopic biopsy. CONCLUSION: Surgery remains the treatment of choice for this condition. Intraoperative frozen pathological sectioning helped the surgeon decide the extent of surgery, and radical surgery was avoided. Excision of benign lesions occupying the appendiceal orifice may be indicated for the prevention of later development of acute appendicitis. The prognosis is generally good, with only one of the 26 reported patients complicated with acute appendicitis, who subsequently succumbed due to severe comorbidities and sepsis.

Immunoexpression and prognostic role of hTERT and cyclin D1 in urothelial carcinoma.

The aim was to investigate the expression of human telomerase reverse transcriptase (hTERT) and cyclin D1 in correlation with clinicopathologic features of urothelial carcinoma (UC). Tissue microarrays (TMA) were constructed from paraffin-embedded specimens of 94 UC patients and immunohistochemical staining was used. High hTERT expression was found in 50 (53%) of the 94 tumors and was significantly associated with tumor invasiveness and tumor grade (P=0.008 and 0.0190, respectively). High cyclin D1 expression was found in 69 (73%) of the 94 tumors and was significantly associated with non-invasiveness and smaller tumor size, but there was no correlation with tumor grade. Kaplan-Meier analysis indicated that patients with low hTERT and high cyclin D1 levels had longer local recurrence-free survival (P=0.0482 and 0.0123, respectively). In addition, patients with high cyclin D1 levels had longer disease-free survival (P=0.0195). In conclusion, this study demonstrated that hTERT and cyclin D1 may be of recurrence predictive value for UC, thus providing clinicians with ancillary information when deciding on suitable therapeutic strategies in UC.

Cytologic diagnosis of primary effusion lymphoma in an HIV-negative patient.

Primary effusion lymphoma (PEL) is an unusual and rare type of non-Hodgkin's lymphoma characterized by lymphomatous effusion of pleural, pericardial or peritoneal cavities without lymphadenopathy or organomegaly. It is associated with human herpes virus-8 (HHV-8) and occurs most often in immunodeficient patients. We present a case of PEL in a 69-year-old male presenting with pleural effusion and ascites. Fluid aspiration showed a monomorphic population of atypical lymphoid cells, which were medium- to large-sized, with mono- or binucleated hyperchromatic nuclei and a small to moderate amount of basophilic cytoplasm containing cytoplasmic vesicles. Immunohistochemically, the lymphoid cells expressed CD138 and multiple myeloma oncogene 1, were positive for HHV-8, and were monoclonal for immunoglobulin heavy chain gene rearrangement. They were negative for Epstein-Barr virus by in situ hybridization. Unfortunately, the patient died during the first course of chemotherapy with cyclophosphamide, vincristine and prednisone.

Application of the microarray technique to cell blocks.

OBJECTIVE: To evaluate whether the technique ofmicroarray construction can be applied to paraffin-embedded cell block specinmens. STUDY DESIGN: We used a manual microarray method for construction of a well-aligned cell microarray. First we evaluated the cellularity of the cell block and assigned the cases to the null, low, moderate or high cellularity category. Second, based on the different cellularity, we constructed 25 specimen cylinders into 1 block. We used routine hematoxylin-eosin (H-E) stain and immunocytochemistry (ICC) on the slide. RESULTS: All cell microarray paraffin block shaping and specimen cylinder arraying were finished in 1 step, so the specimen cylinders and the paraffin blocks of the cell microarray could very easily be incorporated. No sample was lost during the H-E staining process. However, a few samples fell off partially during the ICC process. Additionally, we observed that the higher cellularity groups yielded a better outcome as compared to the lower cellularity groups. CONCLUSION: This study introduced a very simple and economical technique for the creation of cell microarrays from cell blocks. This procedure should acquaint cytopathologists with microarray technology and encourage its use.

Use of caveolin-1, thyroid transcription factor-1, and cytokeratins 7 and 20 in discriminating between primary and secondary pulmonary adenocarcinoma from breast or colonic origin.

The objectives of this study were firstly to compare the immunostaining patterns of antibodies against caveolin-1, thyroid transcription factor-1 (TTF-1), cytokeratin 7 (CK7) and cytokeratin 20 (CK20) in primary and secondary pulmonary adenocarcinomas of breast or colonic origin, and secondly, to investigate their use alone and in combination, in distinguishing between primary and secondary lung adenocarcinomas from breast or colonic origin. Of the 49 lung adenocarcinoma specimens that were enrolled in this study, 30 were primary pulmonary adenocarcinomas, and 19 (9, breast origin; 10, colonic origin) were metastatic pulmonary carcinomas. Immunohistochemical staining was used to detect the expression of caveolin-1, TTF-1, CK7, and CK20. Primary pulmonary adenocarcinoma most often had the CK7-positive/CK20-negative immunohistochemical phenotype and was either TTF-1 positive or caveolin-1 negative. Secondary pulmonary adenocarcinoma of breast origin most often had the CK7-positive/CK20-negative immunohistochemical phenotype and was either TTF-1 negative or caveolin-1 positive, while secondary pulmonary adenocarcinoma of colonic origin most often had the CK20-positive/CK7-negative immunohistochemical phenotype and was either TTF-1 negative or caveolin-1 positive. The results suggest that caveolin-1, TTF-1, or CK7/CK20 alone did not distinguish reliably between primary and secondary pulmonary adenocarcinomas originating from breast or colon. The use of a panel of antibodies that includes TTF-1, caveolin-1, and CK7/CK20 may have higher sensitivity in discriminating between primary adenocarcinomas and metastatic lung adenocarcinomas from breast or colonic origin.