Triple-Negative or Close-to-Triple-Negative Breast Cancer Presenting as a Thick-Walled Cystic Lesion.

Clinical and pathologic characteristics of the invasive ductal carcinoma (IDC) presenting as a thick-walled breast cyst are little known. Three female patients were included in this report. A palpable, nontender breast lump was found in all cases. While mammography showed a hyperdense mass, ultrasonography demonstrated a thick-walled cystic mass. Magnetic resonance imaging clearly showed the cystic breast lesions with ring-like or irregular rim enhancement. A grade III IDC was confirmed in all cases. All IDCs but one were estrogen receptor negative, progesterone receptor negative, and human epidermal growth factor receptor 2 negative, with merely weak progesterone receptor positivity (5%) in one case. All cases underwent surgical management first and postoperative chemotherapy. Breast malignancy presenting as a thick-walled cystic mass could be a highly aggressive IDC, even triple-negative breast cancer. It is imperative for breast cancer-related practitioners to identify the potentially malignant cystic lesions timely and adopt appropriate management.

Emerging roles of interactions between ncRNAs and other epigenetic modifications in breast cancer.

Up till the present moment, breast cancer is still the leading cause of cancer-related death in women worldwide. Although the treatment methods and protocols for breast cancer are constantly improving, the long-term prognosis of patients is still not optimistic due to the complex heterogeneity of the disease, multi-organ metastasis, chemotherapy and radiotherapy resistance. As a newly discovered class of non-coding RNAs, ncRNAs play an important role in various cancers. Especially in breast cancer, lncRNAs have received extensive attention and have been confirmed to regulate cancer progression through a variety of pathways. Meanwhile, the study of epigenetic modification, including DNA methylation, RNA methylation and histone modification, has developed rapidly in recent years, which has greatly promoted the attention to the important role of non-coding RNAs in breast cancer. In this review, we carefully and comprehensively describe the interactions between several major classes of epigenetic modifications and ncRNAs, as well as their different subsequent biological effects, and discuss their potential for practical clinical applications.

Up-regulated circBACH2 contributes to cell proliferation, invasion, and migration of triple-negative breast cancer.

An increasing amount of evidence has proven the vital role of circular RNAs (circRNAs) in cancer progression. However, there remains a dearth of knowledge on the function of circRNAs in triple-negative breast cancer (TNBC). Utilizing a circRNA microarray dataset, four circRNAs were identified to be abnormally expressed in TNBC. Among them, circBACH2 was most significantly elevated in TNBC cancerous tissues and its high expression was positively correlated to the malignant progression of TNBC patients. In normal human mammary gland cell line, the overexpression of circBACH2 facilitated epithelial to mesenchymal transition and cell proliferation. In TNBC cell lines, circBACH2 knockdown suppressed the malignant progression of TNBC cells. Mechanistically, circBACH2 sponged miR-186-5p and miR-548c-3p, thus releasing the C-X-C chemokine receptor type 4 (CXCR4) expression. The interference of miR-186-5p/miR-548c-3p efficiently promoted the cell proliferation, migration, and invasion suppressed by circBACH2 knockdown in the TNBC cell lines. Finally, circBACH2 knockdown repressed the growth and lung metastasis of TNBC xenografts in nude mice. In summary, circBACH2 functions as an oncogenic circRNA in TNBC through a novel miR-186-5p/miR-548c-3p/CXCR4 axis.

TRIM3 inhibits P53 signaling in breast cancer cells.

BACKGROUND: Beast cancer is the most common women cancer worldwide, while two third of them are ER alpha positive breast cancer. Among the ER alpha positive breast cancer, about 80% are P53 wild type, indicating the potential tumor suppression role in ER alpha positive breast cancer. Since P53 is an important safeguard to inhibit cell malignant transformation, reactivating P53 signaling could a plausible approach to treat breast cancer. METHODS: TRIM3 protein levels were measured by western blot, while the P53 classical target genes were measured by real-time PCR. WST1 assay were used to measure cell proliferation, while cleaved caspase-3 was used to detect cell apoptosis. Protein stability and ubiquitin assay were used to detect the P53 protein ubiquitin and stability. The immuno-precipitation assays were used to detect the protein interactions. Immuno-staining was used to detect the protein localization of P53 and TRIM3, while the ubiquitin-based immuno-precipitation assays were used to detect the specific ubiquitination manner of P53. RESULTS: In our study, we identified TRIM3 as an endogenous inhibitor for P53 signaling. TRIM3 depletion inhibited breast cancer cell proliferation and promoted apoptosis. In addition, TRIM3 depletion increased P53 protein level in breast cancer cell. Further investigation showed that TRIM3 could associate with P53 and promote P53 K48-linked ubiquitination and degradation. CONCLUSION: Our study identified a novel post-translational modification mechanism between TRIM3 and P53. TRIM3 depletion or blockage could be a promising strategy to rescue P53 signaling and inhibit breast cancer progression.

Circular RNA circ-ZEB1 acts as an oncogene in triple negative breast cancer via sponging miR-448.

BACKGROUND: Circular RNAs (circRNAs) play an important role in tumor development. The miRNA sponge is a common role played by circRNAs in various tumors, including breast cancer. OBJECTIVE: This study aimed to explore the role of circ-ZEB1 in the proliferation and apoptosis of triple negative breast cancer (TNBC) cells. METHODS: The expressions of several circRNAs which were predicted to be bound with miR-448 were detected in 30 clinical TNBC tumor tissues and paired paracancer tissues. The cell counting kit-8 assay was performed to detect the TNBC cell proliferation. The TNBC cell apoptosis was detected using the TUNEL assay. The binding between circ-ZEB1 and miR-448, as well as between miR-448 and eukaryotic elongation factor 2 kinase (eEF2 K), was detected using the RNA pull-down assay and/or the luciferase reporter assay. The effect of circ-ZEB1 knockdown on TNBC tumor growth was detected using the mouse xenograft model. RESULTS: Compared with normal tissues and breast epithelial cells, the expression of circ-ZEB1 was markedly higher in TNBC tumor tissues and tumor cell lines. The small hairpin RNA-mediated circ-ZEB1 knockdown inhibited TNBC cell proliferation and induced cell apoptosis. The RNA pull-down assay and the luciferase reporter assay confirmed the binding between circ-ZEB1 and miR-448, as well as between miR-448 and eEF2 K. The knockdown of circ-ZEB1 was proven to inhibit TNBC cell proliferation and tumor growth via releasing miR-448, and subsequently reducing the expression of the miR-448 target, eEF2 K. CONCLUSION: In conclusion, our findings identified a new functional circ-ZEB1 in TNBC tumorigenesis, and revealed the important regulatory role of circ-ZEB1 via sponging miR-448, providing a novel insight for TNBC pathogenesis.

Lentivirus-mediated CDglyTK gene-modified free flaps by intra-artery perfusion show targeted therapeutic efficacy in rat model of breast cancer.

BACKGROUND: Free flap-mediated gene therapy in the tumor bed following surgical resection is a promising approach in cancer targeted treatment of residual disease. We investigated the selective killing efficacy of a lentivirus-mediated cytosine deaminase-thymidine kinase (CDglyTK) gene in transplanted breast cancer delivered into a free flap by intra-artery perfusion. METHODS: Proliferation, apoptosis, and cell cycle of rat SHZ-88 breast cancer cells transfected with a lentivirus-mediated CD/TK gene were measured following treatment with ganciclovir and 5-flucytosine in vitro. A model of residual disease of breast cancer in a rat superficial inferior epigastric artery (SIEA) flap model was used to study the therapeutic potential of a double suicide CD/TK and prodrug system in vivo. RESULTS: Killing efficacy of the double suicide CD/TK and prodrug system on SHZ-88 cells was mediated by increased apoptosis and cell cycle arrest at the G1 phase with significant bystander effect. Following recombinant lentivirus transfection of rat SIEA flap by intra-artery perfusion, CD/TK gene expression was limited to the flap, and the volume and weight of transplanted tumors were significantly reduced without observable toxicity. CONCLUSIONS: SIEA flaps transfected with a lentivirus-mediated CDglyTK gene by intra-artery perfusion effectively suppress transplanted breast tumor growth without obvious systemic toxic effects in rats.

Septocutaneous perforator mapping and clinical applications of the medial arm flap.

BACKGROUND: The medial arm flap has a long history but remains underused despite providing multiple advantages. We reviewed our experience with using the medial arm flap to clarify the distribution of septocutaneous perforators and its relationship with pedicled flap design. PATIENTS AND METHODS: This retrospective study included 36 consecutive patients who underwent reconstructive surgery with a medial arm flap (42 arms). Septocutaneous perforator mapping was conducted using a refined coordinate system originating at the medial epicondyle, with the y-axis running to the axillary apex. RESULTS: At least three perforators were identified along the medial intermuscular septum of the arm, located densely at 88 ± 8%, 49 ± 9%, and 20 ± 6% of the distance between the medial epicondyle and axillary apex (i.e., arm length), with a prevalence of 95.2%, 100%, and 85.7%, respectively. All arms had at least one medium- or large-sized perforator, and 90.5% of arms had at least two such perforators. Twenty-nine flaps were transferred as pedicled distant flaps for head and neck reconstruction or hand reconstruction, while the remaining 13 were transferred as perforator-based propeller flaps for axillary, elbow, or chest wall reconstruction. Thirty-six flaps underwent pre-transfer expansion. The average flap size was 157.9 ±â€¯64.5 cm2 (range: 40-330 cm2). All flaps were successful, except for one perforator-based flap, which was replaced due to partial loss. Donor site morbidity was minimal. CONCLUSIONS: Given its rich septocutaneous perforator distribution, the medial arm flap can be harvested reliably with versatile design and minimal donor site morbidity, thus deserving more attention in reconstructive surgery.

Mechanical Loading Improves Engineered Tendon Formation with Muscle-Derived Cells: An In Vivo Analysis.

BACKGROUND: The authors' previous study showed that muscle-derived cells could regenerate strong engineered tendon with better tissue structure. However, little was known about the mechanism of neotendon built by muscle-derived cells, and the development and maturation of the cells. The authors hypothesized that mechanical loading modulated this process. The aim of this study was to investigate whether mechanical loading could regulate muscle-derived cell-based engineered tendon formation and maturation. METHODS: Muscle-derived cells were isolated, expanded, and seeded onto polyglycolic acid fibers that formed a cell-scaffold complex. After in vitro culture for 2 weeks, half of them were implanted without loading and the other half were sutured to mouse fascia that could provide a natural dynamic loading. At 12 and 24 weeks after implantation, histologic examinations, ultrastructure, and biomechanical characteristics were evaluated. RESULTS: Gross observation results showed that under mechanical loading, neotendon tissue could be generated with muscle-derived cells and the tissue structure became more mature with the increase of culture time. Well-organized aligned collagen fibers and elongated morphologic cells were observed on histologic examination under mechanical loading. In contrast, the nonload group failed to form neotendon, but formed disorganized fibrous tissue with significantly worse mechanical properties and poor collagen fibril structure. CONCLUSIONS: This study demonstrates that mechanical loading is indispensable in tendon tissue engineering with muscle-derived cells. Although muscle-derived cells have a potential advantage in neotendon regeneration, stress deprivation resulted in a distinctly inferior maturity level of engineered tendon.

Facial Defect Reconstruction Using the True Scarless Pre-Expanded Forehead Flap.

OBJECTIVE: This clinical study describes a reconstructive method for facial soft-tissue defects that uses the pre-expanded forehead flap and minimizes donor site morbidities. METHODS: The surgery was subdivided into 3 stages. First stage, an appropriately sized expander was buried underneath the forehead. Second stage, after adequate inflation of the expander, a forehead flap based on the frontal branches of the superficial temporal artery was raised, and the distal portion of the flap was used to reconstruct the facial defect. The cutaneous pedicle of the flap was designed near the frontal hairline. Third stage, 3 weeks later, the flap pedicle was divided, and the forehead incisional scar was melted into the neoreconstructed hairline. RESULTS: Between July 2010 and December 2016, 16 patients underwent facial defect reconstruction. Etiologies included postburn scar (31%), melanocytic nevus (56%), and hemangioma (13%). The mean size of the defects was 8.78 × 5.06 cm (range, 3 × 2.5 to 15 × 7 cm). The average dimension of the forehead flap was 21.63 × 7.38 cm (range, 12 × 4 to 28 × 10 cm). Fifteen flaps survived without any perfusion-related complications. Venous congestion occurred in 1 flap and gradually subsided without any flap loss. Patients were followed after surgery, ranging from 4 to 48 months. Patients and/or their family members were satisfied with the final aesthetic outcomes. CONCLUSION: Facial defect reconstruction using a pre-expanded forehead flap, with the donor-site incisional scar designed along the hairline, can not only provide sufficient tissue for defect reconstruction, but also maximally reduce donor-site morbidities.

Dynamic Penile Corpora Cavernosa Reconstruction Using Bilateral Innervated Gracilis Muscles: A Preclinical Investigation.

INTRODUCTION: Prosthesis-assisted penile reconstruction has been performed extensively to restore a cosmetically acceptable phallus. However, a large number of patients will undergo revision surgery for various prosthesis-related complications. AIM: To develop a 1-stage prosthesis-free dynamic cavernosa reconstruction method using bilateral innervated gracilis muscles and to investigate the feasibility and reliability of the surgical design. METHODS: 10 fresh cadavers were dissected to assess the availability of bilateral gracilis muscles for functional cavernosa rebuilding. 11 mongrel female dogs were involved in the penile reconstruction surgery. The neophallus consisted of bilateral gracilis muscles as the neo-cavernosa, a right gracilis skin flap as the neourethra, and a lower abdominal flap with an anterior rectus sheath as the skin envelope and neo-tunica albuginea. The function and structure of the neo-phalli were assessed 7 months postoperatively. MAIN OUTCOME MEASURES: The neurovascular pedicle length of the gracilis muscles and the volume of the gracilis venter musculi were measured in the cadaveric investigation. The average dimensions of the canine neo-phalli at rest and during electrostimulated erection were obtained and the muscular fatigue-resistant curve was drawn. Histologic evaluations also were performed. RESULTS: The neurovascular pedicle length and volume of the gracilis muscles were sufficient to yield a nearly normal appearance of the neo-cavernosa in the cadaveric and animal studies. The muscular fatigue-resistant curve demonstrated adequate length, stiffness, and duration of erection of the neo-phalli to accomplish normal coitus. Histologic evaluations showed an intact neourethra and nearly normal muscle structure in the inner layer of the canine neo-cavernosa, except for significantly increased amount of collagen fibers and type I/III collagen ratio in the outer layer of the neo-cavernosa. The percentage of type II (fatigue-prone) muscle fibers did not change significantly. CONCLUSION: Our preclinical investigation proves that corpora cavernosa reconstruction using bilateral innervated gracilis muscles is technically feasible and functionally efficacious. Yin Z, Liu L, Xue B, et al. Dynamic Penile Corpora Cavernosa Reconstruction Using Bilateral Innervated Gracilis Muscles: A Preclinical Investigation. Sex Med 2018;6:162-170.

A novel hybrid 3D-printed titanium scaffold for osteogenesis in a rabbit calvarial defect model.

The aim of this study was to explore an innovative method to improve the osteogenic ability of porous titanium. We used gelatin (Gel) and nano-hydroxyapatite (nHA) to construct micro-scaffolds within the pores of porous titanium alloy. We compared three groups: control, Gel:nHA = 1:0, and Gel:nHA = 1:1. We assessed cell attachment, cell proliferation, and osteogenic (alkaline phosphatase [ALP] and collagen type 1 [Col-1]) and cytoskeletal (Talin) gene and protein expression in MC3T3-E1 cells. We also evaluated osteogenic abilities in a rabbit calvarial defect model. Our results showed that micro-scaffolds can improve new bone formation both in vitro and in vivo. Between the two micro-scaffold groups, the Gel:nHA = 1:1 group exhibited the most satisfactory results. It had a multi-hierarchical pore structure with a mean pore size of 156±86 µm. The Gel:nHA = 1:1 group exhibited significantly higher gene and protein expression of ALP, Col-1, and Talin. This group also exhibited the most new bone volume during in vivo experiments. The 3D micro-scaffold structure was an effective method of porous titanium modification that not only provided appropriate cell growth conditions but may also be used as a carrier of bioactive factors in the future.

Total Cheek Reconstruction Using the Pre-Expanded Medial Arm Flap With Functional and Aesthetic Donor Site Closure.

This clinical study aimed at introducing a reconstructive method for total cheek soft tissue defects using the pre-expanded medial arm flap and to maximally reduce donor site morbidities simultaneously. A retrospective review of data was performed for 3 patients requiring the pre-expanded medial arm flap to reconstruct total cheek defects. The donor site in the medial arm was primarily closed using a pedicle parascapular or thoracodorsal artery perforator flap. All patients had a total unilateral cheek defect after lesion removal. Three pre-expanded medial arm flaps measuring 17 × 11 cm, 22 × 15 cm, and 20 × 15 cm were separately used for cheek defect reconstruction. The donor sites in the medial arm were closed using a pedicle parascapular flap, measuring 22 × 9.5 cm, and 2 pedicle thoracodorsal artery perforator flaps, measuring 22 × 10 cm and 23 × 10 cm. No major complications occurred in any patient. Patients were followed up for 3, 12, and 18 months. Patients (and/or their family members) were satisfied with the final outcomes. Total cheek defect reconstruction using a pre-expanded medial arm flap, with the donor site closed using a pedicle axial back flap, can not only provide sufficient tissue for cheek resurfacing, but also guarantee primary donor site closure and could be an alternative reconstructive option for patients who have a total cheek defect.

[Functional penile reconstruction with skin flaps combined with muscle: a preclinical study].

Objective: To devAop a new approach for functional penile reconstruction. Methods: Total penile reconstruction using innervated gracilis muscles and anterolateral thigh (ALT) flap with fascia lata was performed on four human cadavers. Canine models were constructed using gracilis muscles and lower abdominal flap with anterior rectus sheaih, instead of the ALT flap. Seven months later, the erectile function of the neophalli elicited by electrostimulation was evaluated. The length, diameter and stiffness were measured respectively and the results were analyzed statistically using paired-samples t test. Results: The penile reconstruction was success with good appearance in the cadaveric study. The hardness of the reconstructed penis in canine models was graded as 1.2 ± 0.4 at rest. With the nerve pedicles stimulated, it increased significantly and was graded as 3.4 ± 0.5 (P ＜ 0.05),while the length shortened and the diameter did not change significantly. A successful erection-mimic course was observed. Conclusions: It indicates that penile reconstruction with skin flaps combined with muscle is feasible, serving as a theoretical and technical support for further clinical application.