TBOPP, a DOCK1 Inhibitor, Potentiates Cisplatin Efficacy in Breast Cancer by Regulating Twist-mediated EMT.

BACKGROUND: DOCK1 has been reported to be involved in tumor progression and resistance. 1-(2-(30-(trifluoromethyl)-[1,10-biphenyl]-4-yl)-2-oxoethyl)-5-pyrrolidinylsulfonyl2(1H)- pyridone (TBOPP) is a selective DOCK1 inhibitor; however, the role and molecular mechanisms of DOCK1 and its inhibition in breast cancer (BC) resistance remain poorly understood. OBJECTIVE: This study aims toinvestigate the underlying mechanisms of DOCK1 in BC resistance. METHODS: DOCK1 or Twist siRNA and Twist plasmid were used to explore the function of DOCK1 in vitro experiments. A mouse xenograft model was used for in vivo experiments. RESULTS: In the present study, we demonstrated that DOCK1 siRNA promoted cisplatin sensitivity in BC cells. Moreover, TBOPP also enhances the therapeutic effect of cisplatin both in vitro and in vivo. Mechanistically, DOCK1 siRNA inhibited EMT. Twist 1 is one of the EMT-inducing transcription factors and is known to induce EMT. To further reveal the effect of DOCK in BC cells, we co-transfected with DOCK1 and Twist1 siRNA to BC cells and found that co-transfection with DOCK1 and Twist siRNA could not further enhance the cisplatin sensitivity of BC cells. Moreover, DOCK1 siRNA failed to reverse the effect of Twist 1 up-regulation. CONCLUSION: Taken together, these results demonstrate that DOCK1 may function as a potential therapeutic target in BC and that combining cisplatin with TBOPP may provide a promising therapeutic strategy for cisplatin-resistant BC patients.

High-Throughput Microfluidic Production of Droplets and Hydrogel Microspheres through Monolithically Integrated Microchannel Plates.

In this paper, we present a highly effective microfluidic emulsion system using an integrated microchannel plate (MCP), a porous glass membrane that is readily available and densely packs millions of through-microchannels, for high-throughput production of monodisperse droplets. The physical controls of droplet formation, including viscosity, flow rate, and pore size, have been extensively explored for optimum emulsification conditions. The performance of the device has been validated where monodisperse droplets with a narrow coefficient of variance (<5%) can be achieved at a dispersed phase flux of 3 mL h-1 from a piece of 4 × 4 mm2 MCP. The average droplet size is two times the nominal membrane pore diameter and thus can be easily controlled by choosing the appropriate membrane type. The preparation of hydrogel microspheres has also been demonstrated with a high throughput of 1.5 × 106 particles min-1. These microspheres with a uniform size range and rough surface morphology provide suitable bioenvironments and serve as ideal carriers for cell culture. Mouse fibroblasts are shown to be cultured on these 3D scaffolds with an average cell viability of over 96%. The cell attachment rate can reach up to 112 ± 7% in 24 h and the proliferation ability increases with the number of culture days. Furthermore, the device has been applied in the droplet digital polymerase chain reaction for absolute quantification of lung cancer-related PLAU genes. The detection limit achieved was noted to be 0.5 copies/µL with a dynamic range of 105 ranging from 1 × 102 to 1 × 106 copies/µL. Given the easy fabrication, robust performance, and simple operation, the emulsion system sets the stage for the laboratory's droplet-based assays and applications in tissue engineering.

Photoacoustic imaging in monitoring of compartmental syndrome in rat extremities.

Muscle ischemia injury is the essence of compartment syndrome (CS). Photoacoustic (PA) imaging can monitor hemoglobin concentration changes in ischemic tissue by determining the state of light-absorbing molecules. This study investigated whether PA imaging can provide accurate CS monitoring. Rats received compression on the lower hind limb for 3 h to induce ischemia injury, followed by PA imaging of desired muscles for 24 h. PA intensities of the injured group were significantly lower than that in the control group. Histology findings correlated well with the PA findings. The results demonstrated that PA imaging could be a noninvasive and timely tool for clinically monitoring CS.

miR-137 alleviates doxorubicin resistance in breast cancer through inhibition of epithelial-mesenchymal transition by targeting DUSP4.

Acquired resistance to chemotherapy is a major obstacle in breast cancer (BC) treatment. Accumulated evidence has uncovered that microRNAs (miRNAs) are vital regulators of chemoresistance in cancer. Growing studies reveal that miR-137 acts as a suppressor in tumor progression. However, it remains obscure the role of miR-137 in modulating the sensitivity of BC cells to doxorubicin (DOX). In this study, we demonstrate that miR-137 exerts a significant effect on repressing the development of chemoresistance of BC cells in response to DOX via attenuating epithelial-mesenchymal transition (EMT) of tumor cells in vitro and in vivo. MiR-137 overexpression dramatically elevated the sensitivity of BC cells to DOX as well as impaired the DOX-promoted EMT of tumor cells. Mechanistically, miR-137 directly targeted dual-specificity phosphatase 4 (DUSP4) to impact on the EMT and chemoresistance of BC cells upon DOX treatment. Consistently, decreased DUSP4 efficiently enhanced the sensitivity of BC cells to DOX while overexpressed DUSP4 significantly diminished the beneficial effect of miR-137 on BC cells chemoresistance. Moreover, the increased miR-137 heightened the sensitivity of BC cells-derived tumors to DOX through targeting DUSP4 in vivo. Together, our results provide a novel insight into the DOX resistance of BC cells and miR-137 may serve as a new promising therapeutic target for overcoming chemoresistance in BC.

Early treatment using fractional CO2 laser before skin suture during scar revision surgery in Asians.

Fractional CO2 laser is one of the most effective treatment options used to resurface scars. However, most previous studies have been performed on mature scars at least 2 months after surgery. Recent studies have emphasized the importance of early treatment to reduce scar formation. In the present study, we described our experience with fractional CO2 laser intervention before skin suture during scar revision surgery in Asians, and found the treatment was safe and effective.

WP1130 increases cisplatin sensitivity through inhibition of usp9x in estrogen receptor-negative breast cancer cells.

Approximately 30% of all breast cancers are caused by a lack of estrogen receptor (ER), which renders the cancer resistant to endocrine-based therapy. Many studies suggest that ubiquitin-specific peptidase 9, X-linked (usp9x) regulates multiple cellular behaviors, such as tumor growth, invasion, and resistance to chemotherapeutic agents. This study aimed to evaluate the anti-tumor effects of WP1130, a partially selective inhibitor of deubiquitinating enzymes, in breast cancer cells. We found that WP1130 enhanced cisplatin cytotoxicity in ER-negative tumor cells (MDA-MB-231 and MDA-MB-468), but had little effect in ER-positive Bcap-37 cells. Western blot analysis revealed that usp9x expression was dramatically lower in ER-positive cells compared to that in ER-negative cells. Furthermore, WP1130 treatment suppressed the expression of usp9x and Mcl-1 in ER-negative cells, but not in ER-positive cells. In addition, we found that knockdown of usp9x diminished the chemosensitization activity of WP1130 on breast cancer cells in the presence of cisplatin. Taken together, these results demonstrated that combined treatment with WP1130 could increase the cisplatin sensitivity in a usp9x-dependent manner in estrogen receptor-negative breast cancer cells.

N1-Guanyl-1,7-Diaminoheptane Sensitizes Estrogen Receptor Negative Breast Cancer Cells to Doxorubicin by Preventing Epithelial-Mesenchymal Transition through Inhibition of Eukaryotic Translation Initiation Factor 5A2 Activation.

BACKGROUND: Approximately 30% of breast cancer does not express the estrogen receptor (ER), which is necessary for endocrine-based therapy approaches. Many studies demonstrated that eukaryotic translation initiation factor 5A2 (eIF5A2) serves as a proliferation-related oncogene in tumorigenic processes. METHODS: The present study used cell viability assays, EdU incorporation assays, western blot, and immunofluorescence to explore whether N1-guanyl-1,7-diaminoheptane (GC7), which inhibits eIF5A2 activation, exerts synergistic cytotoxicity with doxorubicin in breast cancer. RESULTS: We found that GC7 enhanced doxorubicin cytotoxicity in ER-negative HCC1937 cells but had little effect in ER-positive MCF-7 and Bcap-37 cells. Administration of GC7 reversed the doxorubicin-induced epithelial-mesenchymal transition (EMT) in ER-negative breast cancer cells. Knockdown of eIF5A2 by siRNA inhibited the doxorubicin-induced EMT in ER-negative HCC1937 cells. CONCLUSION: These data demonstrated that GC7 combination therapy may enhance the therapeutic efficacy of doxorubicin in estrogen negative breast cancer cells by preventing EMT through inhibition of eIF5A2 activation.

G15 sensitizes epithelial breast cancer cells to doxorubicin by preventing epithelial-mesenchymal transition through inhibition of GPR30.

Resistance to single or multiple chemotherapeutic drugs is a major obstacle in breast cancer therapy. Recent studies have suggested that GPR30 is implicated in mediating cancer cell proliferation. The aim of this study was to examine the anti-tumor effects of the GPR30 antagonist G15 in breast cancer. We found that low concentrations of G15 had little effect on breast cancer cell viability, but could enhance doxorubicin sensitivity in MDA-MB-231 and MCF-7 cells with epithelial phenotypes. In addition, G15 prevented epithelial breast cancer cells undergoing epithelial-mesenchymal transition (EMT) after doxorubicin induction. Moreover, downregulation of GPR30 suppressed the EMT in breast cancer cells. These results support that G15 enhanced doxorubicin sensitivity and prevented the EMT in epithelial breast cancer cells by inhibiting GPR30 expression.

microRNA-375 inhibits osteogenic differentiation by targeting runt-related transcription factor 2.

microRNAs (miRNAs) are involved in the regulation of almost all physiological and pathological processes, including osteoblast proliferation and differentiation. miR-375 has been reported to be aberrantly expressed in various tumor types and to inhibit cell proliferation and invasion by targeting a number of key genes. However, the effects of miR-375 on osteogenic differentiation have not been investigated. Thus, the present study aimed to elucidate the function served by miR-375 in osteogenic differentiation. The expression levels of miR-375 were observed to decrease in a C2C12 cell model of osteogenic differentiation. Overexpression of miR-375 inhibited the activity of key osteoblast markers, including osteocalcin (OC), alkaline phosphatase (ALP) and collagen, type I, α 1 (COL1A1). By contrast, inhibition of miR-375 expression resulted in an increase in the osteogenic potential, as indicated by the enhanced expression levels of OC, ALP and COL1A1. In addition, a dual-luciferase reporter assay indicated that runt-related transcription factor 2 (RUNX2) was a target of miR-375. Western blot analysis revealed that the inhibition of miR-375 led to a significant increase in the protein expression levels of RUNX2. In addition, overexpression of RUNX2 was observed to attenuate the miR-375-mediated suppression of osteogenic differentiation. Therefore, the results demonstrated that miR-375 was able to inhibit osteogenic differentiation via the regulation of RUNX2 expression.

Axillary Osmidrosis Treatment Using an Aggressive Suction-Curettage Technique: A Clinical Study on Paired Control.

BACKGROUND: Axillary osmidrosis results in social and psychological problems. Liposuction-assisted techniques used as treatments have been reported to have high recurrence rates. This study aims at introducing the aggressive suction-curettage technique in comparison to the open excision procedure. METHODS: From February 2009 to February 2014, 130 patients were randomly divided into two groups (experimental and control groups). The experimental group was treated with aggressive suction-curettage, while the control group was treated with an open excision. Postoperative assessment (skin necrosis, hematoma, scars and malodour recurrence) and patient satisfaction were surveyed and analyzed. RESULTS: A lower necrosis rate was observed in patients who received aggressive suction-curettage (1.88 %, p < 0.01), higher patient satisfaction (33.85 %, p < 0.01), higher recurrence rate (11.54 %, p < 0.05), and less ugly scars (0.77 %, p < 0.01), compared to those who had the open excision procedure. CONCLUSIONS: The aggressive suction-curettage technique appears to be a reliable treatment option for axillary osmidrosis.

EIF5A2 is a novel chemoresistance gene in breast cancer.

BACKGROUND: The eIF5A2 gene (encoding the eukaryotic initiation factor 5A2) located at 3q26 is a putative oncogene that is overexpressed in colon and rectal carcinomas, lung cancer and hepatocellular carcinoma. EIF5A2 overexpression correlates significantly with tumor metastasis and is an adverse prognostic marker. However, eIF-5A2 overexpression in breast cancer and its effect on chemotherapy are unknown. METHODS: We measured eIF-5A2 expression and doxorubicin sensitivity in different human breast cancer cell lines (Bcap-1937, HCC1937, and MCF-7). To investigate a role for eIF-5A2 in chemoresistance, cells were treated with eIF-5A2-siRNA, exposed to various concentrations of doxorubicin, and toxicity was assayed by CCK-8 (cell counting kit). RESULTS: The eIF-5A2 expression levels varied among breast cancer cells. Higher expression levels correlated with decreased doxorubicin sensitivity. Silencing of eIF-5A2 significantly improved doxorubicin toxicity in all three breast cancer cell lines. CONCLUSION: This study shows that eIF-5A2 plays an important role in doxorubicin chemoresistance in breast cancer cells.

MicroRNA-185 inhibits proliferation by targeting c-Met in human breast cancer cells.

MicroRNAs (miRNAs) are a group of small non-coding RNA molecules that have been shown to regulate the expression of genes involved in tumorigenesis. The relevance of miRNAs in the development, progression and prognosis of human breast cancer is not fully understood. miR-185 has been demonstrated to be involved in the pathogenesis of several types of cancers; however, its role in breast cancer has not yet been elucidated. In the present study, the expression of miR-185 was analyzed by quantitative polymerase chain reaction. In addition, an MTT assay and flow cytometry were used to determine the rates of cell proliferation and apoptosis. Protein expression was analyzed by western blotting and the target gene was confirmed using a luciferase reporter assay. The expression of miR-185 was found to be downregulated in the breast cancer tissues. The MTT assay revealed that overexpression of miR-185 inhibited the proliferation of MDF7 and SKBR3 cells. Furthermore, flow cytometric analysis demonstrated that increased expression levels of miR-185 promoted the apoptosis of breast cancer cells. In addition, c-Met expression was demonstrated to be significantly upregulated in breast cancer tissues and cells, and the c-Met gene was identified to be a target of miR-185. Therefore, the results demonstrated that miR-185 inhibited the proliferation of breast cancer cells by regulating the expression of c-Met, indicating its potential as a therapeutic target for breast cancer.

Tanshinone IIA blocks epithelial-mesenchymal transition through HIF-1α downregulation, reversing hypoxia-induced chemotherapy resistance in breast cancer cell lines.

The aim of the present study was to investigate the effects of tanshinone IIA (Tan IIA), an active constituent of Salvia miltiorrhiza Bunge, on epithelial-mesenchymal transition (EMT) and hypoxia-induced chemoresistance in breast cancer cells. To induce hypoxia, MCF-7 and HCC1973 cells were treated with 100 µM deferoxamine followed by doxorubicin (DOX). Cell viability and proliferation were examined using the CCK-8 and EdU assays, respectively. Western blot and immunofluorescence analyses of the expression of two EMT markers, E-cadherin and vimentin, were also carried out. The role of HIF-1α and TWIST in mediating the effects of Tan IIA was determined through siRNA. Based on the results, hypoxia-induced DOX resistance was observed in both MCF-7 and HCC1973 cells (both P=0.001), which was reversed with Tan IIA. Specifically, in hypoxic conditions, Tan IIA significantly decreased cell viability and proliferation (all P≤0.001), but not apoptosis. Hypoxia also significantly reduced E-cadherin and increased vimentin protein levels (P≤0.005), which returned to control levels with Tan IIA. In addition, silencing both HIF-1α and TWIST expression abrogated the effects of Tan IIA on cell viability. Taken together, Tan IIA ameliorated hypoxia-induced DOX resistance and EMT in breast cancer cell lines, which may be attributed to the downregulation of HIF-1α expression. Further in vivo studies, however, are required to fully elucidate the therapeutic potential of Tan IIA in increasing the sensitivity of breast cancer cells to chemotherapy.

Knockdown of dual specificity phosphatase 4 enhances the chemosensitivity of MCF-7 and MCF-7/ADR breast cancer cells to doxorubicin.

BACKGROUND: Breast cancer is the major cause of cancer-related deaths in females world-wide. Doxorubicin-based therapy has limited efficacy in breast cancer due to drug resistance, which has been shown to be associated with the epithelial-to-mesenchymal transition (EMT). However, the molecular mechanisms linking the EMT and drug resistance in breast cancer cells remain unclear. Dual specificity phosphatase 4 (DUSP4), a member of the dual specificity phosphatase family, is associated with cellular proliferation and differentiation; however, its role in breast cancer progression is controversial. METHODS: We used cell viability assays, Western blotting and immunofluorescent staining, combined with siRNA interference, to evaluate chemoresistance and the EMT in MCF-7 and adriamycin-resistant MCF-7/ADR breast cancer cells, and investigate the underlying mechanisms. RESULTS: Knockdown of DUSP4 significantly increased the chemosensitivity of MCF-7 and MCF-7/ADR breast cancer cells to doxorubicin, and MCF-7/ADR cells which expressed high levels of DUSP4 had a mesenchymal phenotype. Furthermore, knockdown of DUSP4 reversed the EMT in MCF-7/ADR cells, as demonstrated by upregulation of epithelial biomarkers and downregulation of mesenchymal biomarkers, and also increased the chemosensitivity of MCF-7/ADR cells to doxorubicin. CONCLUSIONS: DUSP4 might represent a potential drug target for inhibiting drug resistance and regulating the process of the EMT during the treatment of breast cancer.

Removal of a large amount of pretarsal tissue through three mini incisions in the construction of a double eyelid.

BACKGROUND: The surgical procedure aimed at constructing the upper eyelid crease is the cosmetic operation most commonly requested by Asian patients. The incisional approach leaves a permanent crease, but the swelling period can last longer than 3 months, and a distinct scar usually is unavoidable. Many mini-incisional techniques with incisions of varying sizes and locations have been developed, but the removal of pretarsal tissue has not been sufficient because these procedures wipe out only small pockets of soft tissue immediately inferior to skin for placement of the suture. Thus, the formed double eyelid may not be durable. The technique the authors introduce is a modified mini-incisional technique that combines the benefits of both the incisional and the usual mini-incisional methods. METHODS: Three 3- to 4-mm mini-incisions were made on each upper eyelid. The orbicularis muscle was isolated carefully from the skin and then cut off as much as possible in three directions: down (toward the palpebral margin), left, and right. Finally, the three incisions were sutured, and a vivid fold was created. This technique was applied for 110 patients. The follow-up period ranged from 3 to 12 months. The patients were welcomed for further treatment if any problems occurred at any time. An extra 3 years of follow-up evaluation was applied for 24 patients (46 cases) to assess the long-term maintenance of the fold. RESULTS: This study enrolled 110 patients (212 cases). A natural-looking skin fold appeared after the edema period. The scar was not obvious. Disappearance of the fold was not found in any case during 3-12 months (mean, 9 months) of follow-up evaluation. Obvious shifting of the fold was found in four cases during the additional 3 years of follow-up evaluation, but disappearance of the fold was not found in any case. CONCLUSION: This modified technique was found to be successful in developing a natural-appearing, long-lasting suprapalpebral fold with an inconspicuous scar. It combines the best of both the open and usual mini-incisional techniques. Because a large amount of pretarsal soft tissue is removed, the double fold is less likely to fade away. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article.

[Fisheries biology of Nemipterus bathybius and its optimum catchable size in northern South China Sea].

Based on the investigation data in 1997-1999 and 1964-1965, this paper studied the population structure, stock density distribution and its seasonal variation, growth and mortality, as well as the feeding behaviour of Nemipterus bathybius (Snyber) in northern South China Sea. The results indicated that the body length of the samples was 4.2-21.5 cm, and body weight was 2.9-241 g, with the length of 9.6-14.0 cm and weight of 27 -59 g as predominant. The stock density was the highest (9.92 kg km(-2)) in spring and the lowest (5.53 kg x km(-2)) in summer, and had an obvious zonal distribution. Catch rate increased with the water depth in 60-150 m water zone. Macrura, fishes and cephalopods were the main foods of Nemipterus bathybius. The growth parameters of von Bertalanffy formula estimated with ELEFAN I in FiSAT II package were Linfinity = 22.39 cm, k = 0.44 a(-1), and t0 = -0.63 a. The natural mortality calculated with Pauly's empirical equation was M = 0.94, and the total mortality estimated by length-converted catch curve was Z = 2.65. Assessment with Beverton-Holt model indicated that the optimum catchable age and body length were 1.1 a and 12 cm, respectively. The stock was over-exploited, mainly due to the capture of under-size juveniles. It was suggested that the legal body length of Nemipterus bathybiu at its first capture in northern South China Sea should be 12 cm.