A New Surgical Method of Correcting Abnormal Cartilage in Mild or Moderate Cryptotia.

OBJECTIVE: This study aimed to introduce a new technique for the correction of mild or moderate types of cryptotia. METHODS: During January 2010 to March 2015, patients with cryptotia deformities came to our hospital and were enrolled in this study. A new surgical method of correction of abnormal cartilage by combining irregular incisions in auricle malformation and directional transplantation of auricular cartilage was designed. The irregular Z shaped flap was designed to correct the skin defects. In the ipsilateral cavum conchae, the auricular cartilage strut was harvested, followed with irregular cartilage incisions and cartilage transplantation. RESULTS: After correction, the corrected auricles in 3 cases showed natural contour with deep auriculotemporal sulcus and no conspicuous scars. The antihelix folding and backward rotation deformities of the auricular cartilage were corrected, and the morphology of superior and inferior crus of the antihelix was not destroyed. One case appeared necrotic. During 1 year of follow-up, there was no incidence of recurrence or revision. CONCLUSIONS: The combined techniques for correction of abnormal cartilage are simple and easy to operate, and the reconstructed auricles show natural contour without conspicuous scars. The new technique is suitable for correction of mild and moderate cryptotia.

Validation of Moorong Self-Efficacy Scale among Chinese stroke survivors.

PURPOSE: To examine the psychometric properties of a Chinese version of the Moorong Self-Efficacy Scale (MSES-C) among stroke survivors. METHODS: A cross-sectional descriptive study was conducted with 160 stroke survivors recruited from the three neurology departments in China. Reliability, concurrent validity, and construct validity of the scale were determined. RESULTS: The MSES-C whole scale showed good internal consistency with a Cronbach's α of 0.953. There was a moderate to a strong positive correlation between the MSES-C and Chinese version of the General Self-Efficacy Scale (r = 0.695, p < 0.000), a strong positive correlation between the MSES-C and Chinese version of the stroke specific self-efficacy scale (r = 0.801, p < 0.000), positive correlations between the MSES-C and Chinese versions of the Modified Barthel Index (r = 0.695, p < 0.000), and a negative correlation between the MSES-C and National Institutes of Health Stroke Scale (r = -0.511, p < 0.000). Known-group validity was also supported. CONCLUSIONS: The MSES-C is a reliable and valid instrument for assessing self-efficacy in Chinese stroke survivors.Implications for RehabilitationThe Chinese version of the Moorong Self-Efficacy Scale demonstrated good internal consistency and showed satisfactory concurrent and construct validity among stroke survivors.The Moorong Self-Efficacy Scale can be used to assess stroke recovery among the Chinese population in clinical and research settings.

Preparation of thioamides from alkyl bromides, nitriles, and hydrogen sulfide through a thio-Ritter-type reaction.

A novel thio-Ritter-type reaction of alkyl bromides, nitriles, and hydrogen sulfide has been explored, providing a straightforward approach toward functionally important thioamides. This transformation features a broad substrate scope, operational simplicity, use of available feedstock chemicals, and late-stage functionalizations of bioactive molecules. The reaction mechanism is also proposed.

Comparison of small-diameter decellularized scaffolds from the aorta and carotid artery of pigs.

AIM: Tissue-specific extracellular matrix promotes tissue regeneration and repair. We aimed to identify the optimal decellularized matrices for tissue-engineered vascular graft (TEVG). METHODS: Decellularized aorta of fetal pigs (DAFP, n = 6, group A), decellularized aorta of adult pigs (DAAP, n = 6, group B), and decellularized carotid artery of adult pigs (DCAP, n = 6, group C) were prepared. Scaffolds were compared using histology and ultrastructure. Endothelial cell (EC) and myofibroblast (MFB) infiltration assessments were performed in vitro. Cell infiltration was measured in vivo. Biomechanical properties were also determined. RESULTS: Almost original cells were removed by the acellularization procedure, while the construction of the matrix basically remained. In vitro, monolayer ECs and multi-layer MFBs were formed onto the internal surface of the specimens after 3 weeks. In vivo, cell infiltration in group A significantly increased at the 6th and 8th week when compared with groups B and C (p < 0.01). The infiltrated cells were mainly MFBs and a few CD4+ T-lymphocytes/macrophages in the specimens. Groups A and B showed greater axial compliance than group C (p < 0.01). CONCLUSION: DAFP was the most suitable for use as a small-caliber vascular graft.

The organocatalytic enantiodivergent fluorination of ß-ketodiaryl-phosphine oxides for the construction of carbon-fluorine quaternary stereocenters.

Commercially available cinchona alkaloids that can catalyze the enantiodivergent fluorination of ß-ketodiarylphosphine oxides were developed to construct carbon-fluorine quaternary stereocenters. This protocol features a wide scope of substrates and excellent enantioselectivities, and it is scalable.

S100P contributes to chemosensitivity of human ovarian cancer cell line OVCAR3.

S100P is a member of the S100 family of calcium-binding proteins. Our previous studies have demonstrated its significant downregulation in oxaliplatin-resistant colon cancer cell line. The present study investigated whether it plays a role in the regulation of chemosensitivity to anticancer drugs using human ovarian cancer cell line OVCAR3. We firstly overexpressed S100P in the OVCAR3 cell line, and evaluated the expression level of S100P by semiquantitative RT-PCR, Western blotting and immunofluorescence assay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay indicated that overexpression of S100P sensitized OVCAR3 cells for chemotherapeutic drugs (paclitaxel, oxaliplatin, 5-fluorouracil, etoposide and epirubicin) induced cytotoxicity more than vector-only controls. Further studies showed that downregulation of S100P by RNA interference in OVCAR3 cells led to a significant increase of resistance to each of these anticancer drugs. Taken together, our results suggest that S100P plays an important role in regulation of chemosensitivity to anticancer drugs in ovarian cancer cells. Using S100P as a molecular biomarker may increase our ability to predict tumor drug response in ovarian cancer.

Decellularized aorta of fetal pigs as a potential scaffold for small diameter tissue engineered vascular graft.

BACKGROUND: For cardiovascular tissue engineering, acellularized biomaterials from pig have been widely investigated. Our purpose was to study mechanical properties and biocompatibility of decellularized aorta of fetal pigs (DAFP) to determine its potential as scaffold for small diameter tissue engineered vascular graft. METHODS: Descending aorta of fetal pigs was removed cells using trypsin, ribonuclease and desoxyribonuclease. Mechanical properties of DAFP were evaluated by tensile stress-strain and burst pressure analysis. Assessment of cell adhesion and compatibility was conducted by seeding porcine aortic endothelial cells. To evaluate biocompatibility in vivo, DAFP was implanted subcutaneously into adult male Sprague Dawley rats for 2, 4 and 8 weeks. RESULTS: Histochemistry and scanning electron microscopy examination of DAFP revealed well-preserved extracellular matrix proteins and porous three-dimensional structures. Compared with fresh aorta, DAFP had similar ultimate tensile strength, axial compliance and burst pressure. Cell culture studies in vitro showed that porcine aortic endothelial cells adhered and proliferated on the surfaces of DAFP with excellent cell viability. Subdermal implantation demonstrated that the DAFP did not show almost any immunological reaction and exhibited minimal calcification during the whole follow-up period. CONCLUSION: The DAFP has the potential to serve as scaffolds for small diameter tissue engineered vascular graft.

Low expression of S100P associated with paclitaxel resistance in ovarian cancer cell line.

BACKGROUND: Recent studies indicate that S100P expression may be a biomarker that can predict the success of cancer chemotherapy. Whether it is relevant to chemotherapeutics in ovarian cancer is unknown. In this study, we investigated the association of S100P expression with paclitaxel sensitivity in ovarian cancer cell lines. METHODS: We measured S100P expression and paclitaxel resistance profiles in parent SKOV3 and OVCAR3 cell lines. Then, the two cell lines were transiently transfected with S100P siRNA. We also constructed an OVCAR3 cell clone that stably overexpressed S100P. The effect of S100P expression level on the survival of cells exposed to paclitaxel was measured using the MTT assay. S100P expression was evaluated by semi-quantitative RT-PCR and Western blotting. Significance of differences was calculated using independent samples t-test and one way analysis of variance (ANOVA). RESULTS: Lower S100P expression was associated with a survival advantage in OVCAR3 cells exposed to paclitaxel; the survival advantage in SKOV3 cells was smaller (P < 0.05). The survival advantage associated with decreased S100P expression was even greater for SKOV3 and OVCAR3 cells that had been transfected with S100P siRNA before being exposed to paclitaxel (P < 0.05). Consistent with this, the OVCAR3 cell clone that was transfected to overexpress S100P was more sensitive to paclitaxel (P < 0.05). CONCLUSIONS: Low S100P expression contributes to drug resistance to paclitaxel in ovarian cancer cell lines. S100P expression thus might be a marker that can predict the effectiveness of paclitaxel based chemotherapy. Such a marker could be helpful in improving individual medication regimens for ovarian cancer patients.

Expression profiles analysis of long non-coding RNAs identified novel lncRNA biomarkers with predictive value in outcome of cutaneous melanoma.

Recent advancements in cancer biology have identified a large number of lncRNAs that are dysregulated expression in the development and tumorigenesis of cancers, highlighting the importance of lncRNAs as a key player for human cancers. However, the prognostic value of lncRNAs still remains unclear and needs to be further investigated. In the present study, we aim to assess the prognostic value of lncRNAs in cutaneous melanoma by integrated lncRNA expression profiles from TCGA database and matched clinical information from a large cohort of patients with cutaneous melanoma. We finally identified a set of six lncRNAs that are significantly associated with survival of patients with cutaneous melanoma. A linear combination of six lncRNAs (LINC01260, HCP5, PIGBOS1, RP11-247L20.4, CTA-292E10.6 and CTB-113P19.5) was constructed as a six-lncRNA signature which classified patients of training cohort into the high-risk group and low-risk group with significantly different survival time. The prognostic value of the six-lncRNA signature was validated in both the validation cohort and entire TCGA cohort. Moreover, the six-lncRNA signature is independent of known clinic-pathological factors by multivariate Cox regression analysis and demonstrated good performance for predicting three- and five-year overall survival by time-dependent receiver operating characteristic (ROC) analysis. Our study provides novel insights into the molecular heterogeneity of cutaneous melanoma and also shows potentially important implications of lncRNAs for prognosis and therapy for cutaneous melanoma.

Development and in vivo validation of tissue-engineered, small-diameter vascular grafts from decellularized aortae of fetal pigs and canine vascular endothelial cells.

BACKGROUND: Tissue engineering has emerged as a promising alternative for small-diameter vascular grafts. The aim of this study was to determine the feasibility of using decellularized aortae of fetal pigs (DAFPs) to construct tissue-engineered, small-diameter vascular grafts and to test the performance and application of DAFPs as vascular tissue-engineered scaffolds in the canine arterial system. METHODS: DAFPs were prepared by continuous enzymatic digestion. Canine vascular endothelial cells (ECs) were seeded onto DAFPs in vitro and then the vascular grafts were cultured in a custom-designed vascular bioreactor system for 7 days of dynamic culture following 3 days of static culture. The grafts were then transplanted into the common carotid artery of the same seven dogs from which ECs had been derived (two grafts were prepared for each dog with one as a backup; therefore, a total of 14 tissue-engineered blood vessels were prepared). At 1, 3, and 6 months post-transplantation, ultrasonography and contrast-enhanced computed tomography (CT) were used to check the patency of the grafts. Additionally, vascular grafts were sampled for histological and electron microscopic examination. RESULTS: Tissue-engineered, small-diameter vascular grafts can be successfully constructed using DAFPs and canine vascular ECs. Ultrasonographic and CT test results confirmed that implanted vascular grafts displayed good patency with no obvious thrombi. Six months after implantation, the grafts had been remodeled and exhibited a similar structure to normal arteries. Immunohistochemical staining showed that cells had evenly infiltrated the tunica media and were identified as muscular fibroblasts. Scanning electron microscopy showed that the graft possessed a complete cell layer, and the internal cells of the graft were confirmed to be ECs by transmission electron microscopy. CONCLUSIONS: Tissue-engineered, small-diameter vascular grafts constructed using DAFPs and canine vascular ECs can be successfully transplanted to replace the canine common carotid artery. This investigation potentially paves the way for solving a problem of considerable clinical need, i.e., the requirement for small-diameter vascular grafts.

Molecular cloning, expression, bioinformatics analysis and bioactivity characterization of TNF13B (BAFF) gene in bat (Vespertilio superans Thomas).

B cell activating factor (BAFF) belonging to the tumor necrosis factor (TNF) family is critical to B cell survival, proliferation, maturation, and immunoglobulin secretion and to T cell activation. In the present study, the full-length cDNA of BAFF (designated bBAFF) from the bat (Vespertilio superans Thomas) was cloned using RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of bBAFF consists of 986 bases including an 873 bp open reading frame encoding 290 amino acids. Sequence comparison indicated that the amino acid of bBAFF possessed the TNF signature, a transmembrane domain, a putative furin protease cleavage site and three cysteine residues. The predicted three-dimensional (3D) structure of the bsBAFF monomer, analyzed by comparative protein modeling, revealed that it was very similar to its counterparts. Real-time quantitative PCR (qPCR) analysis indicated that bBAFF mRNA was predominantly expressed in bat lymphoid tissue spleen. The SUMO (Small Ubiquitin-like Modifier)-bsBAFF was efficiently expressed in Escherichia coliBL21 (DE3) and confirmed by SDS-PAGE and Western blotting analysis. Laser scanning confocal microscopy analysis showed that bsBAFF could bind to its receptors on B cells. In vitro, the MTT assays indicated that SUMO-bsBAFF was not only able to promote survival/proliferation of bat lymphocytes but also able to stimulate survival/proliferation of mouse B cells. These findings indicate that bsBAFF plays an important role in the survival/proliferation of B cells and has functional cross-reactivity among mammalians. The present findings may provide valuable information for research into the immune system of the bat.

Selection of single chain variable fragments specific for the human-inducible costimulator using ribosome display.

We applied a ribosome display technique to a mouse single chain variable fragment (scFv) library to select scFvs specific for the inducible costimulator (ICOS). mRNA was isolated from the spleens of BALB/c mice immunized with ICOS protein. Heavy and κ chain genes (VH and κ) were amplified separately by reverse transcriptase polymerase chain reaction, and the anti-ICOS VH/κ chain ribosome display library was constructed with a special flexible linker by overlap extension PCR. The VH/κ chain library was transcribed and translated in vitro using a rabbit reticulocyte lysate system. Then, antibody-ribosome-mRNA complexes were produced and panned against ICOS protein under appropriate conditions. However, in order to isolate specific scFvs for ICOS, negative selection using CD28 was carried out before three rounds of positive selection on ICOS. After three rounds of panning, the selected scFv DNAs were cloned into pET43.1a and detected by SDS-PAGE. Then, enzyme-linked immunosorbent assay showed that we successfully constructed a native ribosome display library, and among seven clones, clone 5 had the highest affinity for the ICOS and low for the CD28. Anti-ICOS scFvs are assessed for binding specificity and affinity and may provide the potential for development of the humanized and acute and chronic allograft rejection.

Molecular cloning, bioinformatics analysis and functional characterization of B-cell activating factor in goat (Capra hircus).

B-cell activating factor of the TNF family (BAFF) induces B cell survival, proliferation, immunoglobulin secretion and has a role in enhancing immune responses. In the present study, we amplified the cDNA of goat (Capra hircus) BAFF (designated gBAFF) from spleen by reverse transcription-PCR (RT-PCR). The open reading frame (ORF) of gBAFF covers 843 bp encoding 280 amino acids, with a 152-aa mature peptide. Sequence comparison indicated that the amino acid of gBAFF possessed the TNF signature, a transmembrane domain, a putative furin protease cleavage site and three cysteine residues. The predicted three-dimensional (3D) structure of the soluble part of gBAFF (gsBAFF) analyzed by "comparative protein modelling" revealed that it was very similar to its counterparts. Real-time quantitative PCR (qPCR) analysis indicated that gBAFF mRNA was predominantly expressed in goat lymphoid tissue spleen. Recombinant gsBAFF was fused with a small ubiquitin-related modifier (SUMO) gene to enhance the soluble expression level in Escherichia coli BL21 (DE3). The resulting fused protein SUMO-gsBAFF was efficiently expressed and purified using metal chelate affinity chromatography (Ni-NTA), then confirmed by SDS-PAGE and Western blotting analysis. In vitro, the MTT assays and flow cytometric analysis indicated that SUMO-gsBAFF as well as gsBAFF could promote the survival/proliferation of goat splenic B cells or mouse splenic B cells. Therefore, BAFF may be a potential immunologic factor for enhancing immunological efficacy in goat.

S100P sensitizes ovarian cancer cells to carboplatin and paclitaxel in vitro.

OBJECTIVE: To investigate the relationship between the S100P and the sensitivity of ovarian cancer to chemotherapeutics. METHOD: We established stable cell lines of ovarian cancer cells, SKOV3 and OVCAR3, that overexpress human S100P. We also transiently transfected the parent cell lines with S100P-targeted siRNA for down-regulation of S100P expression. The sensitivity of all transfected and untransfected cell lines to carboplatin and paclitaxel was detected by MTT assay. RESULTS: For both cells, IC50s decreased to carboplatin and paclitaxel (p<0.05), with overexpression of S100P compared to untransfected cells. Alternatively, with down-regulation of S100P by siRNA, the IC50 to carboplatin and paclitaxel increased in each case (p<0.05), which was significantly higher compared to untransfected cells. CONCLUSION: Changes in expression levels of S100P in SKOV3 and OVCAR3 cells results in variable susceptibility to carboplatin and paclitaxel. These data suggest that S100P contributes to chemosensitivity to carboplatin and paclitaxel in ovarian cancer cells.