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Eutectic In-48Sn was considered a promising candidate for low-temperature solder due to its low melting point and excellent mechanical properties. Both Cu2(In,Sn) and Cu(In,Sn)2 formation were observed at the In-48Sn/Cu interface after 160 °C soldering. However, traditional mechanical polishing produces many defects at the In-48Sn/Cu interface, which may affect the accuracy of interfacial reaction investigations. In this study, cryogenic broad Ar+ beam ion milling was used to investigate the interfacial reaction between In-48Sn and Cu during soldering. The phase Cu6(Sn,In)5 was confirmed as the only intermetallic compound formed during 150 °C soldering, while Cu(In,Sn)2 formation was proven to be caused by room-temperature aging after soldering. Both the Cu6(Sn,In)5 and Cu(In,Sn)2 phases were confirmed by EPMA quantitative analysis and TEM selected area electron diffraction. The microstructure evolution and growth mechanism of Cu6(Sn,In)5 during soldering were proposed. In addition, the Young’s modulus and hardness of Cu6(Sn,In)5 were determined to be 119.04 ± 3.94 GPa and 6.28 ± 0.13 GPa, respectively, suggesting that the doping of In in Cu6(Sn,In)5 has almost no effect on Young’s modulus and hardness.

Background Related to the pathogenesis of cancers in humans, the interaction between erythropoietin-producing hepatocyte receptors and ephrins (Ephs/ephrins) affects and regulates various biological functions. Erythropoietin-producing hepatocyte receptor type A2 (EphA2) is a tyrosine kinase that binds to ephrins (e.g., ephrin-A1) to initiate bidirectional signaling between cells. The binding of EphA2 and ephrin-A1 leads to the inhibition of Ras-MAPK activity and tumor growth. During tumorigenesis, the normal interaction between EphA2 and ephrin-A1 is hindered, which leads to the overexpression of EphA2 and induces cancer. The overexpression of EphA2 has been identified as a notable tumor marker in the diagnosis and treatment of pancreatic cancer. Results In this study, we used phage display to isolate specific antibodies against the active site of EphA2 molecules by using a discontinuous recombinant epitope for immunization. The therapeutic efficacy and inhibition mechanism of the generated antibody against pancreatic cancer was validated and clarified. The generated antibodies were bound to the conformational epitope of endogenous EphA2 on cancer cells, thus inducing cellular endocytosis and causing EphA2 degradation. Molecule signals pAKT, pERK, pFAK, and pSTAT3 were weakened, thereby inhibiting the proliferation and migration of pancreatic cancer cells. The humanized antibody hSD5 could effectively inhibit the growth of the xenograft pancreatic cancer tumor cells BxPc-3 and Mia PaCa-2 in mice, respectively. When antibody hSD5 was administered in combination with gemcitabine, significantly synergistic effects on tumor growth inhibition (reach 79.3%) were observed. Conclusions On the basis of the efficacy of the IgG hSD5 antibody, clinical administration of the hSD5 antibody is likely to suppress tumors in patients with pancreatic cancer and abnormal activation or overexpression of EphA2 signaling.

For highly conserved mammalian protein, chicken is a suitable immune host to generate antibodies. Monoclonal antibodies have been successfully targeted with immunity checkpoint proteins as a means of cancer treatment; this treatment enhances tumor-specific immunity responses through immunoregulation. Studies have identified the importance of B7-H4 in immunoregulation and its use as a potential target for cancer treatment. High levels of B7-H4 expression are found in tumor tissues and are associated with adverse clinical and pathological characteristics. Using the phage display technique, this study isolated specific single-chain antibody fragments (scFvs) against B7-H4 from chickens. Our experiment proved that B7-H4 clearly induced the inhibition of T-cell activation. Therefore, use of anti-B7-H4 scFvs can effectively block the exhaustion of immunity cells and also stimulate and activate T-cells in peripheral blood mononuclear cells. Sequence analysis revealed that two isolated scFv S2 and S4 have the same VH complementarity-determining regions (CDRs) sequence. Molecule docking was employed to simulate the complex structures of scFv with B7-H4 to analyze the interaction. Our findings revealed that both scFvs employed CDR-H1 and CDR-H3 as main driving forces and had strong binding effects with the B7-H4. The affinity of scFv S2 was better because the CDR-L2 loop of the scFv S2 had three more hydrogen bond interactions with B7-H4. The results of this experiment suggest the usefulness of B7-H4 as a target for immunity checkpoints; the isolated B7-H4-specific chicken antibodies have the potential for use in future cancer immunotherapy applications.

The use of scaled-down micro-bumps in miniaturized consumer electronic products has led to the easy realization of full intermetallic solder bumps owing to the completion of the wetting layer. However, the direct contact of the intermetallic compounds (IMCs) with the adhesion layer may pose serious reliability concerns. In this study, the terminal reaction of the Ti adhesion layer with Cu–Sn IMCs was investigated by aging the micro-bumps at 200 °C. Although all of the micro-bumps transformed into intermetallic structures after aging, they exhibited a strong attachment to the Ti adhesion layer, which differs significantly from the Cr system where spalling of IMCs occurred during the solid-state reaction. Moreover, the difference in the diffusion rates between Cu and Sn might have induced void formation during aging. These voids progressed to the center of the bump through the depleting Cu layer. However, they neither affected the attachment between the IMCs and the adhesion layer nor reduced the strength of the bumps. In conclusion, the IMCs demonstrated better adhesive behavior with the Ti adhesion layer when compared to Cr, which has been used in previous studies.

BackgroundA reliable cancer biomarker will be critical for advanced colorectal cancer (CRC) therapeutic approaches since current treatments are limited to certain patient characteristics, such as age, sex, comorbidities and patients received self-expandable metal stents implantations. The association of epidermal growth factor-like domain 6 (EGFL6) with cancer development has been reported. Here, we focused on the role of EGFL6 in CRC progression and its clinical relevance. MethodsAn anti-EGFL6 antibody was generated by phage display technology to investigate the potential therapeutic efficacy in CRC. Methylene blue staining was applied to investigate the EGFL6 expression in CRC patients and animal tissue. Protein and DNA level of EGFL6 expression as well as related pathway investigation were determined by western blot and quantitative real time PCR. Silence EGFL6 by siRNA transfection, transwell migration and invasion assay were performed to verify EGFL6 function. Student t test, Kruskal-Wallis test and multiple comparisons were used to statistical analysis results. ResultsSignificant EGFL6 expression was found in colon tissues from patients and spontaneous tumorigenesis mouse but not in normal tissue. Furthermore, we found EGFL6 could enhance cancer cell migration, invasion and proliferation in CRC via up-regulating ERK/ AKT pathway, as well as reducing ADAMTS1 and Snail expression. We also found EGFL6 regulates cell abilities through EGFR/αvβ3 integrin receptors. By conducting animal experiments, our anti-EGFL6 antibody, EGFL6-E5-IgG, showed tumor inhibition and anti-metastasis ability. Furthermore, no impact on angiogenesis and wound healing by using EGFL6-E5-IgG were observed. ConclusionsWe demonstrated that EGFL6 plays a role in CRC tumorigenesis and tumor progression, indicating that EGFL6 is a potential cancer biomarker and therapeutic target worth further investigation.

The Astragalus membranaceus polysaccharides (APS) can improve immunity and enhance treatment reactions. This study analyzed the effects of effective antivascular endothelial growth factor (anti-VEGF) antibody production in mice treated with APS. After APS treatment, the serum of mice produced the antibody reactions that can cross-validate VEGF. The isolated single-chain fragment variable (scFv) antibodies could neutralize VEGF and inhibit in vivo tumor growth. Of the scFvs, scFv 4E can significantly compete the interaction of bevacizumab with VEGF. In cell experiments, scFv 4E effectively inhibited human umbilical vein endothelial cells induced by VEGF in vitro. In a matrix gel-assisted angiogenesis model, scFv 4E significantly inhibited angiogenesis reactions. In addition, in a xenograft model established in the colorectal cancer cell strain HCT116, scFv 4E treatment inhibited tumor growth by up to 52.7%. Finally, molecule docking was performed to simulate the complex interactions of scFv 4E and VEGF, the main driving forces of which involve the hydrophobic interactions and hydrogen bonds of Tyr108 and Tyr 109 of the complementarity-determining region H3 loop with VEGF. The results help in establishing antibody library with high diversity for selecting antibodies with specificity. In addition, this study indirectly expounded the correlations of APS enhancing immunity regulation in vivo.

BACKGROUND The aberrant glycosylation on proteins and lipids has been implicated in malignant transformations for promoting the tumorigenesis, metastasis, and evasion from the host immunity. The I-branching β-1,6-N-acetylglucosaminyltransferase, converting the straight i to branched I histo-blood group antigens, reportedly could influence the migration, invasion, and metastasis of solid tumors. STUDY DESIGN AND METHODS We first chose the highly cytotoxic natural killer (NK)-92MI cells as effector against leukemia for this cell line has been used in several clinical trials. Fluorescence-activated cell sorting and nonradioactive cytotoxicity assay were performed to reexamine the role of NK-activating receptors, their corresponding ligands, and the tumor-associated carbohydrate antigens in this NK-92MI-leukemia in vitro system. The I role on cytotoxic mechanism was further studied especially on the effector-target interactions by cytotoxic analysis and conjugate formation assay. RESULTS We showed that expression levels of leukemia surface ligands for NK-activating receptors did not positively reflect susceptibility to NK-92MI. Instead, the expression of I antigen on the leukemia cells was found important in mediating the susceptibility to NK targeting by affecting the interaction with effector cells. Furthermore, susceptibility was shown to dramatically increase while overexpressing branched I antigens on the I- cells. By both conjugate and cytotoxicity assay, we revealed that the presence of I antigen on leukemia cells enhanced the interaction with NK-92MI cells, increasing susceptibility to cell-mediated lysis. CONCLUSION In our system, branched I antigens on the leukemia were involved in the immunosurveillance mediated by NK cells specifically through affecting the effector-target interaction.

BACKGROUND Phosphorylation status of the transcription factor CCAAT/enhancer binding protein α (C/EBPα) has been demonstrated in a human hematopoietic cell model to regulate the formation of branched I antigen by affecting its binding affinity to the promoter region of the IGnTC gene during erythroid and granulocytic differentiation. STUDY DESIGN AND METHODS The K-562 cell line was induced to differentiate into red blood cells (RBCs) or granulocytes by sodium butyrate or retinoic acid, respectively, to study the involvement of three MAP kinase pathways in I antigen synthesis. The regulatory effects of the extracellular signal-regulated kinase (ERK)2-Src homology region 2 domain-containing phosphatase 2 (SHP2) pathway on phosphorylation status and binding affinities of C/EBPα as well as the subsequent activation of IGnTC and synthesis of surface I formation were studied in wild-type K-562 cells and in mutant cells that overexpress ERK2 and SHP2. RESULTS We found that SHP2-ERK2 signaling regulates the phosphorylation status of C/EBPα to alter its binding affinity onto the IGnTC promoter region, thereby activating the synthesis of cell surface I antigen formation during erythropoiesis. CONCLUSION SHP2-ERK2 signaling acts upstream of C/EBPα as a regulator of cell surface I antigen synthesis. Such regulation is specific for RBC but not for granulocyte differentiation.

The human cluster of differentiation 19 (CD19) is highly expressed in most leukemia, rendering is a promising therapeutic target. In this study, we generated anti-CD19 single-chain variable fragments (scFv) from immunized chickens by phage display technology. After constructing a scFv antibody library with 2.5 × 10

To understand the mechanism for inhibition of hepatitis B virus (HBV) infection is important. In this study, single-chain variable fragment (scFv) antibodies were generated and directed to the pre-S2 epitope of HBV surface antigen (HBsAg). These human scFvs were isolated from a person with history of HBV infection by phage display technology. An evaluation of panning efficiency revealed that the eluted phage titer was increased, indicating that specific clones were enriched after panning. Selected scFvs were characterized with the recombinant HBsAg through Western blotting and enzyme-linked immunosorbent assay to confirm the binding ability. Flow cytometry analysis and immunocytochemical staining revealed that one scFv, S17, could recognize endogenous HBsAg expressed on the HepG2215 cell membrane. Moreover, the binding affinity of scFv S17 to the pre-S2 epitope was determined to be 4.2 × 10-8 M. Two ion interactions were observed as the major driving forces for scFv S17 interacting with pre-S2 by performing a rational molecular docking analysis. This study provides insights into the structural basis to understand the interactions between an antibody and the pre-S2 epitope. The functional scFv format can potentially be used in future immunotherapeutic applications.

The aberrant glycosylation on proteins and lipids has been implicated in malignant transformations for promoting the tumorigenesis, metastasis, and evasion from the host immunity. The I-branching β-1,6-N-acetylglucosaminyltransferase, converting the straight i to branched I histo-blood group antigens, reportedly could influence the migration, invasion, and metastasis of solid tumors.We first chose the highly cytotoxic natural killer (NK)-92MI cells as effector against leukemia for this cell line has been used in several clinical trials. Fluorescence-activated cell sorting and nonradioactive cytotoxicity assay were performed to reexamine the role of NK-activating receptors, their corresponding ligands, and the tumor-associated carbohydrate antigens in this NK-92MI-leukemia in vitro system. The I role on cytotoxic mechanism was further studied especially on the effector-target interactions by cytotoxic analysis and conjugate formation assay.We showed that expression levels of leukemia surface ligands for NK-activating receptors did not positively reflect susceptibility to NK-92MI. Instead, the expression of I antigen on the leukemia cells was found important in mediating the susceptibility to NK targeting by affecting the interaction with effector cells. Furthermore, susceptibility was shown to dramatically increase while overexpressing branched I antigens on the I- cells. By both conjugate and cytotoxicity assay, we revealed that the presence of I antigen on leukemia cells enhanced the interaction with NK-92MI cells, increasing susceptibility to cell-mediated lysis.In our system, branched I antigens on the leukemia were involved in the immunosurveillance mediated by NK cells specifically through affecting the effector-target interaction.

Phosphorylation status of the transcription factor CCAAT/enhancer binding protein α (C/EBPα) has been demonstrated in a human hematopoietic cell model to regulate the formation of branched I antigen by affecting its binding affinity to the promoter region of the IGnTC gene during erythroid and granulocytic differentiation.The K-562 cell line was induced to differentiate into red blood cells (RBCs) or granulocytes by sodium butyrate or retinoic acid, respectively, to study the involvement of three MAP kinase pathways in I antigen synthesis. The regulatory effects of the extracellular signal-regulated kinase (ERK)2-Src homology region 2 domain-containing phosphatase 2 (SHP2) pathway on phosphorylation status and binding affinities of C/EBPα as well as the subsequent activation of IGnTC and synthesis of surface I formation were studied in wild-type K-562 cells and in mutant cells that overexpress ERK2 and SHP2.We found that SHP2-ERK2 signaling regulates the phosphorylation status of C/EBPα to alter its binding affinity onto the IGnTC promoter region, thereby activating the synthesis of cell surface I antigen formation during erythropoiesis.SHP2-ERK2 signaling acts upstream of C/EBPα as a regulator of cell surface I antigen synthesis. Such regulation is specific for RBC but not for granulocyte differentiation.

Vestibular disorder is the cause of approximately 50% of dizziness in older people. The vestibular system is a critical postural control mechanism, and posturography analysis is helpful for diagnosing vestibular disorder. In clinical practice, the sensory organization test (SOT) is used to quantify postural control in an upright stance under different test conditions. However, both aging and vestibular disorder cause declines of postural control mechanisms. The aim of this study was to enhance the performance of the SOT using a nonlinear algorithm of empirical mode decomposition (EMD) and to verify the differences of effects caused by aging and/or illnesses benefits to clinical diagnosis. A total of 51 subjects belonging to 3 groups--healthy-young, healthy-elderly and dizzy--were recruited for this study. New dynamic parameters of the SOT were derived from the center of pressure (COP) signals. EMD served as an adaptive filter bank to derive the low- and high-frequency components of the COP. The effects on four ratios of sensory analysis caused by aging and vestibular disorder can be investigated for the specific frequency bands. According to our findings, new SOT parameters derived from the component with the specific frequency band more sensitively reflect the functional condition of vestibular dysfunction. Furthermore, both aging and vestibular dysfunction caused an increase in magnitude for the low-frequency component of the AP-direction COP time series. In summary, the low-frequency fluctuation reflects the stability of postural control, while the high-frequency fluctuation is sensitive to the functional condition of the sensory system. EMD successfully improved the accuracy of SOT measurements in this investigation.

The objective of this research was the establishment of a quality scale [QFD] for creative and product design service that was not available before. The cultural and creative product design industry of Sanxia indigo dyeing was taken as the case study to develop such a Quality Function Deployment (QFD) system. In addition, digital inject textile printing technology was utilized to replicate and analyze the pure indigo blue color and totems used in the traditional indigo dyeing process. As the local cultural and creative industry are progressing rapidly in Taiwan and global markets, this research intends to develop the Old Street of the Sanxia indigo dyeing creative product design industry into a case study, with the objective of exploring the establishment of a quality control scale for a design service entity that has never had one. This research also intends to preserve and record traditional Sanxia indigo craftsmanship and totems with digital printing technology so that it can be scientifically preserved and analyzed. The Sanxia indigo shops and consumers were targeted as the objects of study. This research has probed into the actual operations of the Sanxia indigo dyeing creative product design industry. Data were generated through review and study of domestic and international literature and articles. Interview surveys and on-site observations were conducted, as well as hands-on experience in the shops, to gain a real understanding of the industry. Based on the QFD which originated in Japan in the 1960's, the "voice of the customer" was collected and analyzed for product design and production in order to actually meet or even exceed customer expectations of the products. While the QFD was mainly used for the manufacturing industry, the goal was to try to set up QFD concepts for the design service industry, the Sanxia indigo dyeing creative product design in this case. The House of Quality deployment model was used to conduct a series of matrix analyses, and through the quality deployment mechanism, the "voice of the customer" was transformed into a design scale for the development of creative products. The six foundation frameworks of the House of Quality (Hauser 1988) were explored. The frameworks are: 1. customer demand, 2. engineering analysis, 3. competition analysis, 4. relationship analysis, 5. technology assessment and 6. correlation matrix. The findings will provide industry vendors with important Sanxia indigo dyeing references for product design and quality principles to help meet consumer needs. Three conclusions from this research were obtained: (1) The importance of creative product design quality is reflected in the following four aspects: a. It is a key element of the customer purchasing intention-product feature; b. It is a rule for industry existence and success; c. It can enhance customer satisfaction and loyalty and d. It can improve the operating performance of the industry. (2) If creative industry service procedures are included in the quality scale of creative indigo dyeing product design, a complete quality scale of creative product design can be constructed. (3) Creative indigo dyeing product design quality of Sanxia indigo dyeing art should be strengthened in terms of dye liquor materials, indigo dyeing features and post processing techniques so as to ensure the quality of creative product design and enhance customer satisfaction and purchasing intentions.