The clinical relevance of epithelial-mesenchymal transition and its correlations with tumorigenic immune infiltrates in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a cancer with extremely high mortality. Epithelial-mesenchymal transition (EMT) may play an important role in the occurrence, invasion and prognosis of HCC; however, its relationship with immunity in HCC has not yet been studied. Therefore, we investigated the diagnostic and prognostic values of EMT and explored its potential connections with tumorigenic immune infiltrates in HCC. We first proposed a quantitative metric of EMT activity, the EMT score. After applying this metric to 20 datasets from the Integrative Molecular Database of Hepatocellular Carcinoma, the Cancer Genome Atlas, and the Gene Expression Omnibus, we explored the ability of the EMT score to stratify across sample types. We then applied the EMT score for survival analysis and to differentiate patients with/without vascular invasion to test its prognostic value. We also collected and calculated data on the abundance of immune cells and immune cell markers in HCC and investigated their correlations with EMT scores. Finally, we synthesized and analyzed 20 datasets and constructed an EMT-gene-immune linkage network. The results showed higher EMT scores in HCC samples than in cirrhotic and normal livers. The cases with higher EMT scores also showed poorer performance in terms of prognostic factors such as vascular invasion and overall survival time. Our research demonstrated a broad correlation between EMT and the tumor immune microenvironment, and we uncovered multiple potential linkers associated with both EMT and immunity. Studying EMT has clinical relevance and high diagnostic and prognostic value for HCC.

High level of RNF187 contributes to the progression and drug resistance of osteosarcoma.

Objectives: Ring finger protein 187 (RNF187) was recently demonstrated to be up-regulation and function as a promoter in multiple cancers. However, the roles of RNF187 in osteosarcoma (OS) are unclear. Here, we tried to reveal the clinicopathological and biological roles of RNF187 in OS. Materials and Methods: We employed the quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) to determine the expression of RNF187 in OS tissues and cells. Migration and invasion capacities were analyzed by wound healing and transwell assays, and colony formation and CCK8 assays were performed to investigate proliferative ability. The functional effects of RNF187 on OS drugs resistance were further determined by CCK8 and western blot assays. Then, the relationship between RNF187 expression and clinical implications was analyzed by tissue microarrays (TMAs) including 51 OS cases. Moreover, the prognostic value was also determined by Kaplan-Meier analysis. Results: We reported that RNF187 mRNA was significantly increased in OS tissues compared to matched nontumorous tissues (3.83 ±0.79 vs. 1.70 ± 0.63), which was in line with the IHC assay in TMAs. By RNA interference and cDNA transfection, we showed high level of RNF187 increased the migration, invasion and proliferation of OS cells. Moreover, we demonstrated that elevated RNF187 expression induced OS cell drugs resistance, activated the ERK1/2 molecular and markedly enhanced the BCL-2 expression. Clinically, OS patients with high level of RNF187 was associated with Histologic differentiation (p=0.001), an advanced Enneking stage (p=0.001), response to chemotherapy (p=0.004), and metastasis (p= 0.001). Clinically, our data displayed that the RNF187 overexpression in OS samples associated with shorten overall survival (p=0.001) and high tumor recurrence (p=0.001) in postoperative OS patients. Conclusions: Our results indicate that Elevated RNF187 expression is a new adverse outcomes marker for OS patients and may be used as a new therapeutic target of OS.

Deep lymphatic anatomy of the upper limb: An anatomical study and clinical implications.

BACKGROUND: The deep and perforating lymphatic anatomy of the upper limb still remains the least described in medical literature. MATERIALS AND METHODS: Six upper limbs with the axillary tissue were harvested from three unembalmed human cadavers amputated at the shoulder joint. A small amount of 6% hydrogen peroxide was employed to detect the lymphatic vessels around the deep palmar arch, radial and ulnar neurovascular bundles. A 30-gauge needle was inserted into the vessels and they were injected with a barium sulphate compound. Each specimen was dissected, photographed and radiographed to demonstrate deep lymphatic distribution of the upper limb. RESULTS: Continuing from the deep lymph vessels of the hand, single or multiple deep collecting lymph vessels have been found along the radial, ulnar, anterior and posterior interosseous neurovascular bundles in the forearm, brachial and deep branchial neurovascular bundles in the upper arm. During their courses, lymph nodes were found setting in the trunk of the radial, ulnar and brachial lymph vessels near or in the cubital fossa, and in the axillar. Perforating lymph vessels have been found near the wrist and in the cubital fossa, which linked the superficial and deep lymph vessels. The direction of lymphatic drainage was from the deep to superficial or superficial to deep vessels. CONCLUSION: The deep lymphatic anatomy of the upper limb has been described. The results will provide an anatomical basis for clinical management, educational reference and scientific research.

The deep lymphatic anatomy of the hand.

BACKGROUND: The deep lymphatic anatomy of the hand still remains the least described in medical literature. METHODS: Eight hands were harvested from four nonembalmed human cadavers amputated above the wrist. A small amount of 6% hydrogen peroxide was employed to detect the lymphatic vessels around the superficial and deep palmar vascular arches, in webs from the index to little fingers, the thenar and hypothenar areas. A 30-gauge needle was inserted into the vessels and injected with a barium sulphate compound. Each specimen was dissected, photographed and radiographed to demonstrate deep lymphatic distribution of the hand. RESULTS: Five groups of deep collecting lymph vessels were found in the hand: superficial palmar arch lymph vessel (SPALV); deep palmar arch lymph vessel (DPALV); thenar lymph vessel (TLV); hypothenar lymph vessel (HTLV); deep finger web lymph vessel (DFWLV). Each group of vessels drained in different directions first, then all turned and ran towards the wrist in different layers. CONCLUSION: The deep lymphatic drainage of the hand has been presented. The results will provide an anatomical basis for clinical management, educational reference and scientific research.

Perforating and deep lymphatic vessels in the knee region: an anatomical study and clinical implications.

BACKGROUND: To determine the relationship between the perforating and deep lymphatic vessels in the knee region for clinical implications. METHODS: Four lower limbs from two unembalmed human cadavers were used. Under a surgical microscope, 6% hydrogen peroxide was employed to detect lymph vessels accompanying the small saphenous vein, anterior tibial, posterior tibial and fibular blood vessels all commencing from distal ends of specimens. Each lymphatic vessel was inserted by a 30-gauge needle and injected with a barium sulphate mixture. Each specimen was dissected, radiographed and photographed to determine the perforating and deep lymphatic vessels in the region. RESULTS: A perforating lymph vessel was observed in the popliteal fossa of each specimen. It arose from the superficial popliteal lymph node and terminated in the deep popliteal lymph node. The anterior tibial, posterior tibial and peroneal lymph vessels were discovered in the region travelling with the corresponding vascular bundles. After penetrating the vascular aperture of the interosseous membrane between the tibia and fibula, the anterior tibial lymph vessel entered directly into the deep popliteal lymph node or converged to either the posterior tibial or fibular lymph vessel, before entering the node. The posterior tibial and peroneal lymph vessels entered the deep popliteal lymph node. The efferent lymph vessel of the deep popliteal lymph node travelled with the femoral vascular bundle. CONCLUSION: The perforating and deep lymphatic vessels in the knee region has been presented and discussed. The information advances our anatomical knowledge and the results will benefit clinical management.

MicroRNA-506 is up-regulated in the development of pancreatic ductal adenocarcinoma and is associated with attenuated disease progression.

BACKGROUND: MicroRNA-506 (miR-506) has been reported to function in several tumors as a tumor suppressor gene or oncogene. However, the expression and role of miR-506 in pancreatic ductal adenocarcinoma (PDAC) remains unclear. In this study, we aimed to evaluate the phenotype of miR-506 in PDAC. METHODS: Using miRNA in situ hybridization, we examined the expression of miR-506 in 113 PDACs and 87 paired normal pancreatic tissues. We evaluated miR-506 expression in PDAC cells, normal pancreatic ducts, and acinus/islands, and we analyzed the associations between miR-506 expression and the clinicopathologic characteristics of PDAC patients. RESULTS: miR-506 expression was higher in PDAC than in matched normal pancreatic ductal cells (P < 0.001). On the other hand, the combined group of well and moderately differentiated PDACs showed higher levels of miR-506 than the poorly differentiated ones (P = 0.023). Moreover, miR-506 expression was negatively associated with pathologic T category (P = 0.004) and lymph node metastasis (P = 0.033), suggesting that miR-506 might inhibit the progression of PDAC. CONCLUSIONS: Our results suggest that miR-506 either plays a role as an oncogene in the tumorigenesis and a tumor suppressor in the progression or serves as a house-keeping, tumor-suppressing miRNA, whose expression can be activated by oncogenic signals in early development to hinder the progression of PDAC.

A Method for Making a Lymphatic Specimen of the Dorsum of the Hand.

PURPOSE: To make a lymphatic specimen of the dorsum of the hand for educational and clinical purposes. METHODS: A total of four hands from two unembalmed human cadavers were used. Under a surgical microscope, 6% hydrogen peroxide was employed to detect the lymphatic vessels commencing from fingers. A 30-gauge needle was inserted into vessels and injected with a barium sulphate mixture. Each specimen was radiographed, photographed, and dissected to demonstrate the lymphatic vessels on the dorsum of the hand. After being embalmed in 4% formalin for 3 months, specimen was then sealed in a plexiglass container for display. RESULTS: An average of sixteen lymph collecting vessels were found in the subcutaneous tissue of the dorsum of the hand. The diameter of the vessels varied from 0.2 mm to 0.6 mm. Vessels were the continuation of lymph vessels arising from fingers. They travelled meanderingly in the subcutaneous tissue and traversed over or under the veins when they met. CONCLUSION: Actual and accurate lymphatic distributions of the dorsum of the hand have been described and displayed. The information upgrades our anatomical knowledge and the results will be of benefit for the lymphatic education and clinical application.

[Preliminary study of influence of bone tissue from osteonecrosis of femoral head on the proliferation and differentiation of canine bone marrow mesenchymal stem cells].

OBJECTIVE: To examine the effects of bone tissue from osteonecrosis of femoral head on the proliferation and differentiation of canine bone marrow mesenchymal stem cells in vitro culture. METHODS: A canine model of femoral head osteonecrosis was induced by liquid nitrogen freezing. BMSC were isolated from dog ilium bone marrow by a combination of gradient centrifugation and adherent wall culture. Different bone tissues and BMSC were cultivated indirectly in vitro by co-cultured in Transwell plate. According to the culture media, 3 groups were established: blank group (10%FBS/DMEM), control group (10%FBS/DMEM+ bone tissue from natural femoral head) and experimental group (10%FBS/DMEM+ bone tissue from osteonecrosis of femoral head). Cell proliferation was measured by methyl-thiazol tetrazolium (MTT) method. Cell differentiation was examined by alkaline phosphatase (ALP) staining and its concentration examined. Alizarin red staining method was used to study the calcification effects and Oil red O staining method was used to detect if there was fat emergence. RESULTS: As compared with the blank group, the proliferation in the control and experiment groups were significantly promoted after culturing for Days 1, 3 and 5 (P < 0.05). The proliferation of the experiment group was higher than the control group at Day 5 and 7 day (P < 0.05). After a 7-day co-culturing, ALP staining was positive in the control and experiment groups. At Day 7 and 9, the ALP activity in culture fluid was in this order: control group > experiment group > blank group (P < 0.05). Alizarin red staining show control group had the most calcium nodules (12.17 ± 2.48, P < 0.05) and the number of calcium nodules in the experiment group was more than the blank group (P < 0.05). Oil red O staining show there was no fat emergence after 21 days in every group. CONCLUSION: Both natural and osteonecrotic bone tissue of femoral head could promoted the proliferation of canine BMSC and induces them osteogenic differentiation but not lipo differentiation, bone tissue from osteonecrosis of the femoral head had more competence in promoting proliferation but less capability in inducing osteogenic differentiation than natural bone tissue of the femoral head.

Coexpression of Bcl-2 with epithelial-mesenchymal transition regulators is a prognostic indicator in hepatocellular carcinoma.

The anti-apoptosis factor Bcl-2 is known to contribute to tumorigenesis and metastasis. Epithelial-mesenchymal transition (EMT) may also participate in tumor invasion and metastasis. This study investigated the relationship between coexpression profiles of Bcl-2 and EMT regulators in hepatocellular carcinoma (HCC) tumor samples and clinical outcome. The nuclear (Nu) and cytoplasmic (Cyt) expression of Bcl-2 and the EMT regulators Twist-1, Twist-2, and Snail were determined by immunohistochemical staining in tumor tissue isolated from 97 HCC patients. The clinical prognostic values of both individual protein expression and various expression combinations were investigated using univariate and multivariate survival analysis. Results showed that patients with nuclear expression of Bcl-2 had worse clinical outcomes than patients exhibiting cytoplasmic expression. Overall survival was significantly shorter in HCC patients individually expressing Bcl-2-Nu, Twist-1-Nu, Twist-1-Cyt, and Snail (all P<0.05). Patients coexpressing Bcl-2-Nu with Twist-1-Nu, Twist-1-Cyt, Twist-2, or Snail had even worse prognoses than those expressing no biomarker or any one biomarker alone (all P<0.05). Multivariate analysis showed that HCC patients coexpressing Bcl-2-Nu with Twist-1-Cyt had the worst prognosis. This study provides clinical evidence that nuclear expression of Bcl-2 combined with cytoplasmic expression of Twist-1 is a predictor of very poor prognosis in HCC. Coexpression profiles of Bcl-2 and EMT regulators might aid in the selection of the most efficacious therapy for patients with HCC.