[An analysis of breast cancer patients with ultrasound BI-RADS 3 lesions after minimally invasive excision in clinicopathological features and influencing factors of residual tumor].

Objectives: To examine the clinicopathological characteristics and the influencing factors of the residual tumor of patients with Breast Image Report and Data System (BI-RADS) grade 3 lesions diagnosed with malignancy after minimally invasive excision. Methods: In this retrospective case-control study, clinicopathological data of 69 cases, which had been evaluated as BI-RADS 3 lesions by ultrasound (4 151 cases) diagnosed with breast cancer by minimally invasive excision pathology, were analyzed between May 2012 and June 2016 at the Department of Breast Surgery of the Second Hospital of Shandong University and Linyi People's Hospital. All patients were female, aged (43.4±8.2) years (range: 22 to 70 years). Based on residual tumor after minimally invasive excision, patients were classified into two subgroups: tumor residual group (n=39) and non-tumor residual group (n=30). The clinicopathological features between the two groups were compared. The differences in clinicopathological characteristics were compared in different groups using the χ2 test and the t test. Potential variables identified in the univariate analysis and other relevant variables will be analyzed multivarially using Logistic regression models. The Kaplan-Meier method was applied for survival analysis and survival curves. Results: The breast cancer detection rate of ultrasound BI-RADS 3 lesions was 1.66% (69/4 151), and their maximum diameter of the masses was (1.27±0.45) cm (range: 0.5 to 2.3 cm). Among them, the maximum diameter were ≤1 cm in 28 cases and >1 cm in 41 cases. Histopathological results showed carcinoma in situ in 24 cases and invasive carcinoma in 41 cases, positive expression of the estrogen receptor in 47 cases, positive expression of the progesterone receptor in 43 cases, Ki-67 proliferation index elevated in 26 cases. Axillary metastasis positive rate was 10.1% (7/69). Residual tumor after minimally invasive surgery was found in 39 cases (56.5%). Univariate analysis showed that the tumour residual group showed a significantly increased rate of positive expression of the estrogen receptor (91.9%(34/37) vs. 61.9%(13/21), χ2=7.838, P=0.012). In multivariate analysis, the only variable found to significantly affect the residual tumor was the positive expression of the estrogen receptor (OR=16.852, 95%CI: 1.819 to 156.130, P=0.013). The 5-year disease-free survival rate of breast cancer patients with breast ultrasound BI-RADS 3 lesions was 97.1% and the overall survival rate was 98.6%. Conclusions: BI-RADS 3 lesions diagnosed by ultrasound undergoing ultrasound-guided minimally invasive excision have a certain risk of detected malignancy, approximately 1.66%. Patients with positive expression of the estrogen receptor are more likely to develop residual tumor. A secondary operation should be considered to ensure that no tumor residues remain in the cavity.

Molecular characterization, expression, and functional analysis of chicken TRAF6.

Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a crucial adaptor molecule of the interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily, which can trigger downstream signaling cascades involved in innate immunity. The function of TRAF6 has been clarified in mammals but is poorly understood in chicken. In our study, we investigated TRAF6 function in birds, particularly in chicken innate immune responses, by cloning and characterizing chicken TRAF6 (chTRAF6). The full-length coding sequence of chTRAF6 comprised 1638 bp and encoded a 545-amino acid protein, which shares high sequence similarity with TRAF6 of other species and consists of four structurally conserved domains. Quantitative real-time polymerase chain reaction revealed that chTRAF6 was widely expressed in all tested tissues and its expression was induced in chicken embryo fibroblast cells treated with poly(I:C) and poly(dA:dT). Increased expression of chTRAF6 was observed both in vitro and in vivo following infection with Newcastle disease virus in chickens. Taken together, these results suggest that chTRAF6 plays a vital role in host defense against viral infection in chicken.

Molecular cloning and expression analysis of TRAF3 in chicken.

Tumor necrosis factor receptor-associated factor 3 (TRAF3) is a crucial regulator that suppresses c-Jun N-terminal kinase and non-canonical nuclear factor-kB signaling, but facilitates type I interferon production. To determine TRAF3 function in innate immune responses among birds, particularly chicken, we cloned and characterized the chicken TRAF3 gene (chTRAF3) and detected its tissue expression profile in chicken. We also detected the differential expression of chTRAF3 and its downstream gene interferon-ß (IFN-ß) upon different stimuli in primary chicken embryo fibroblast cells. Two chTRAF3 gene products, chTRAF3-1 and chTRAF3-2, can be produced by alternative splicing. The full-length coding sequence of chTRAF3 (chTRAF3-1) was 1704 base pairs and encoded a protein of 567 amino acids with high identity to TRAF3 homologs from mammals and other birds. The deduced amino acid sequence showed typical characteristics of TRAFs, with a RING finger domain, 2 zf-TRAF motifs, and a MATH domain. Quantitative real-time polymerase chain reaction analysis revealed broad expression of chTRAF3 in all detected tissues, with abundant expression in the spleen, thymus, lung, and small intestine. Expression of chTRAF3 was significantly upregulated in a time- and concentration-dependent manner in chicken embryo fibroblast cells challenged with poly I:C or poly dA-dT. Furthermore, chTRAF3 and IFN-ß mRNA expression from chicken embryo fibroblast cells challenged with Newcastle disease virus F48E9 suffered intense suppression compared with Newcastle disease virus Mukteswar infection. Our results indicate that chTRAF3 plays important roles in defending against both RNA and DNA virus infection.

Identification and association of single-nucleotide polymorphisms in gonadotropin-inhibitory hormone (GnIH) gene with egg production traits in Erlang mountainous chickens.

Gonadotropin-inhibitory hormone (GnIH) gene is an important gene in reproduction. In this study, we screened single-nucleotide polymorphisms (SNPs) in the chicken GnIH gene among 204 individuals in Erlang mountainous chickens. We then analyzed the associations between polymorphisms of the GnIH gene and 5 egg production traits in chickens. Five SNPs (T3305C, T3310C, G3403C, G3411A, and T3591C) were detected. Associations between polymorphic loci and age at first egg, body weight at first egg, weight at first egg, egg weight in 300 days, and egg production in 300 days were analyzed using analysis of covariance. The results showed that SNP1, SNP3, and SNP4 had large effects on age at first egg, while SNP5 had a large effect on body weight at first egg; of the effect of the TT genotype was significantly higher than that of CT (P < 0.01). Further analysis show that the highest frequency (0.2353) haplotype H1H1 was associated with the latest age at first egg. The H4H5 haplotype had a positive effect on egg production in 300 days and a negative effect on weight at first egg. We observed no association between the H3H3 haplotype and body weight at first egg.

[Construction of latent membrane protein 2A chimeric antigen receptor-T cells and their lethal effects on nasopharyngeal carcinoma cells].

Objective: To produce latent membrane protein 2A (LMP2A) chimeric antigen receptor (CAR)-T cells and detect the lethal effect of LMP2A CAR-T cells on nasopharyngeal carcinoma (NPC) cells. Methods: The study was conducted from September 2016 to December 2017.Genetic engineering technology was used to construct anti-LMP2A CAR lentiviral expression vector and sequencing was identified. The expression of anti-LMP2A CAR in the 293T cells was confirmed by western blot. CCK8 assay was used to evaluate the cytotoxicity of LMP2A CAR-T cells to NPC cells. ELISA assay was performed to test IL-2 and IFN-γ releasing of activated LMP2A CAR-T cells. The inhibition effect of LMP2A CAR-T cells on NPC xenograft tumor was observed in vivo. Statistical analysis was performed by statistical software SPSS 21.0. Results: The results of PCR and sequencing showed that anti-LMP2A CAR lentiviral expression vector was constructed successfully. The result of western blot indicated the expression of anti-LMP2A CAR in the 293T cells effectively. The results of CCK-8 assay showed that the killing activities of LMP2A CAR-T cells to LV-LMP2A-CNE1 cells were (72.11±9.75)%, (54.65 ±5.42)% and (36.68±3.80)% at 20∶1, 10∶1 and 5∶1 ratio of effective cells to target cells, and had a statistical difference compared to CD19 CAR-T cells and T cells (P<0.05). There was no significant difference in the killing activities of LMP2A CAR-T cells to CNE1 cells compared with CD19 CAR-T cells and T cells. The results of ELISA showed that the content of IL-2 and IFN-γ in the co-culture supernatant of LMP2A CAR-T cells and LV-LMP2A-CNE1 cells was significantly higher than that of LMP2A CAR-T cells and CNE1 cells which had statistical difference (P<0.05); In vivo experiment, the volume of LMP2A CAR-T cell group was (80.3±10.0) mm(3) which was significantly lower than that of the control groups, and the difference was statistically significant (P<0.05). Conclusion: LMP2A CAR-T cells are successfully prepared and have an obvious targeting cytotoxicity on LMP2A-positive NPC cells.

Upconversion Luminescence Properties of Er3+ Doped Yb2Ti20 Nanophosphor by Gd3+ Codoping.

Er3+ doped Yb2Ti207 nanophosphors by Gd3+ codoping with nominal composition of (Er0.05Yb0.95-xGd3+)2Ti2O7 (x = 0, 0.2, 0.4, 0.6, 0.8, and 0.95) have been prepared by sol-gel method. Er3+-Gd3+ codoped Yb2Ti2O7 was characteristic of a typical face-centered cubic crystal phase, and the unit cell parameter increased linearly with the increase of Gd3+ concentration. Under a 976 nm laser diode excitation, both green and red upconversion emissions were observed and the upconversion emissions were enhanced significantly by Gd3+ codoping, showing the strongest green and red emissions at 80 mol% Gd3+ codoping. The intensity ratio of green to red emissions (Igreen/Ired) increased monotonously with the increase of Gd3+ concentration. The energy transfer between Yb3+ and Er3+ and the variation of local crystal field symmetry of Er3+ by the substitution of Yb3+ by Gd3+ ions led to the improvement of upconversion properties of Er3+-Gd3+ codoped Yb2TiO7 nanophosphors.

Optical Temperature Sensing Behavior Through Stark Sublevels Transitions of Green and Red Upconversion Emissions for Er3+-Yb3+-Li+ Codoped TiO2 Phosphors.

The Er3+-Yb3+-Li+ codoped TiO2 phosphors have been prepared by sol-gel method. The green and red upconversion emissions were observed under a 976 nm laser diode excitation, which were ascribed to 2H11/2 --> 4I15/2, 4S3/2(I)/4S3/2(II) -->4I15/2, and 4F9/2(I)/4F9/2(II) -->4I15/2 transitions of Er3+ Stark sublevels. The fluorescence intensity ratios (FIR), which are corresponding to the transitions of 2H11/2/(4S3/2(I)+4S3/2(II))--> 4I5/2, 4S3/2(I)/4S3/2(II) -->4I15/2, and 4F9/2(II)/4F9/2(II) -->4I15/2, have been studied as a function of temperature in the range of 303 673 K. The temperature sensitivities have been calculated at the maximum value of 0.0020 K-1, 0.0015 K-1, and 0.0011 K-1 at the temperatures of 427 K, 350 K, and 273 K for the three coupled energy level transitions, respectively. The Er3+-Yb3+-Li+ codoped Ti02 phosphor with different temperature sensitivities by Stark sublevels indicated that it is a promising material for application in optical temperature sensing at a wide range of temperature.

Anisotropic hyperelastic behavior of soft biological tissues.

Constitutive laws are fundamental to the studies of the mechanically dominated clinical interventions involving soft biological tissues which show a highly anisotropic hyperelastic mechanical properties. The purpose of this paper was to develop an improved constitutive law based on the Holzapfel-Gasser-Ogden's model: to replace the isotropic part with Gent constitutive law so as to model the noncollagenous matrix of the media due to its generality and capability to reproduce the Neo-Hookean model. This model is implemented into an in-house finite element program. A uniaxial tension test is considered to study the influence of material parameter J(m) in Gent model and ß which represents the angle between the collagen fibers and the circumferential direction. A simulation of an adventitial strip specimen under tension is performed to show the applicability of this constitutive law.

[Effects of anti-idiotypic antibody NP30 on modulation of egg granuloma formation and hepatic fibrosis of schistosomiasis].

OBJECTIVE: To study the effects of the monoclonal anti-idiotypic antibody NP30 active immunization on egg granuloma formation and hepatic fibrosis in Schistosoma japonicum infection. METHODS: ICR mice were actively immunized with NP30 100 micrograms x 3 i.p. every 10 days while the mice in control group were injected with SP2/0 ascites i.p. simultaneously. After cercariae challenging, the mice were killed at the 4th, 8th, 12th, 16th, 20th and 24th week, respectively. Mouse livers were removed and stained histochemically with VG and subjected to immunohistochemical assay of collagen type I, III and fibronectin(FN). The volume of egg granulomas and the content of collagen type I, III and FN were determined quantitatively by NYD-1000 Image Analysis System. RESULTS: The volume of egg granulomas in NP30 immunized group was much smaller than that of control group from the 12th week after cercariae challenge. The cellular components of egg granulomas in NP30 immunized group were significantly different from those of the control group, exhibiting two types of atypical egg granulomas were found. VG stain revealed that the average optical density of collagen in hepatic granulomas of experimental group was lower than that of control group. Immunohistochemical assay revealed that the contents of collagen type I, III and fibronectin in egg granulomas of experimental group were lower than those of control group. CONCLUSION: NP30 vaccination may induce both cellular and humoral protective immunity to modulate egg granulomas and suppress liver fibrosis of schistosomiasis japonica.

[Cloning and sequence analysis of the light chain variable region gene of monoclonal anti-idiotypic antibody NP30 of Schistosoma japonicum].

OBJECTIVE: To amplify and sequence the light chain of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum. METHODS: By comparing the conserved regions at each end of the nucleotide sequences of murine germ-line genes encoding FR1 and FR4 regions of immunoglobulin light chain variable regions, we designed a set of primers for amplification of VL gene. The hybridoma cells secreting anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum were cultured and their genome DNAs were extracted and used as templates for PCR. The PCR product was then cloned into pUC19 vector. The recombinants were sequenced by Sanger's method. The VL gene was compared with GenBank and published mouse VL genes. RESULTS: The full-length of VL gene was 318 bp. The VL gene was a member of mouse Ig kappa light chain subgroup IV and generated from rearrangement of germ line V and J kappa 4 genes. The VL gene sequence has been registered by GenBank(accession No. AF206720). CONCLUSION: The obtained VL gene was a potentially functional gene of anti-idiotypic monoclonal antibody NP30 of Schistosoma japonicum.

A novel approach to modelling and simulating the contact behaviour between a human hand model and a deformable object.

A deeper understanding of biomechanical behaviour of human hands becomes fundamental for any human hand-operated activities. The integration of biomechanical knowledge of human hands into product design process starts to play an increasingly important role in developing an ergonomic product-to-user interface for products and systems requiring high level of comfortable and responsive interactions. Generation of such precise and dynamic models can provide scientific evaluation tools to support product and system development through simulation. This type of support is urgently required in many applications such as hand skill training for surgical operations, ergonomic study of a product or system developed and so forth. The aim of this work is to study the contact behaviour between the operators' hand and a hand-held tool or other similar contacts, by developing a novel and precise nonlinear 3D finite element model of the hand and by investigating the contact behaviour through simulation. The contact behaviour is externalised by solving the problem using the bi-potential method. The human body's biomechanical characteristics, such as hand deformity and structural behaviour, have been fully modelled by implementing anisotropic hyperelastic laws. A case study is given to illustrate the effectiveness of the approach.

Stretched inverse opal colloid crystal substrates-induced orientation of fibroblast.

Recently, there has been increasing interest in studying the interaction between mammalian cells and nanometer-sized structures. However, the effect of nanostructures on cell behavior, such as cell morphology and alignment, is still largely unknown. Inverse opal colloid crystal substrates, which can be stretched to produce nano-scale pore structures of different degrees of orientation, serve as a convenient model system to study the effect of nanotopography on cell morphology and cell alignment. In this work, we fabricated inverse opal colloidal crystal films that were either unstretched or stretched to three, four or six times their original length, producing pore structures of increasing degree of orientation. Human dermal fibroblast-fetal (HDF-f) cells were seeded and cultured on these four types of substrates. The results from fluorescence microscopy and scanning electron microscopy indicated that cells showed the highest degree of alignment when cultured on inverse opal colloid crystal films that were stretched the most (six times original length). The results also demonstrated that the orientation of nanostructures could affect both the morphology and growth direction of fibroblasts. The ability to control the direction of cell growth through the engineering of nanostructures could have important applications in tissue engineering, especially for tissues with anisotropic structures, such as cardiac muscle, blood vessel, tendon and ligament.