BACKGROUND: Forest frog is a rare medicinal animal in China, but the skin of forest frog is waste after Oviductus Ranae production. The forest frog skin is rich of collagen, and is large in quantity without pollution and disease risk. So the forest frog skin has potential to be developed into collagen sponge; however, there is no research on collagen sponge preparation as yet.OBJECTIVE: To optimize the preparation of collagen sponge from forest frog skin, and to investigate the physical properties and in vitro cell compatibility.METHODS: Chitosan and glutaraldehyde were used to modify collagen sponge from forest frog skin. Chitosan/collagen (w/w) (1:1, 1:2, 1:4) and glutaraldehyde concentration (1%, 1.5% and 2%) were selected as the experimental factors.The significant water absorbency, mechanical properties and thermal denaturation temperature were chosen as the indexes. Using the orthogonal experimental design, we optimized collagen sponge preparation process. We also investigated the in vitro cell compatibility and surface morphology of the collagen sponge. The nine kinds of collagen sponges from forest frog skins were co-cultured with human foreskin fibroblasts to detect cell proliferation.RESULTS AND CONCLUSION: When the chitosan/collagen was 1:1 and the glutaraldehyde concentration was 1%, we could get the collagen sponge with ideal water absorbency (water absorption capacity=5.22), mechanical properties (elongation at break=10.96%) and thermal denaturation temperature (81.24 ℃). The aperture of the forest frog skin collagen sponge was 200-400 μm, and the pores were consistent in the size and arranged regularly. Except the chitosan/collagen of 1:4 and the glutaraldehyde concentration of 1% or 5%, all kinds of forest frog skin collagen sponges could promote the viability of human foreskin fibroblasts and exerted benefits to cell viability and growth. To conclude,the forest frog skin collagen sponge has good biocompatibility and apparent morphology, in aggreement with the requirements of biological materials.

Objective To investigate the relationship between molecular subtypes of breast cancer and postoperative loco-regional recurrence (LR) in early breast cancer patients with 1-3 positive axillary lymph nodes (pN1) and to improve the individualized indications for post-mastectomy radiotherapy (PMRT)in these patients.Methods A total of 547 patients with pT1-2 N1M0 breast cancer,who received mastectomy between December 1998 and December 2009 in Sun Yat-sen University Cancer Center,were retrospectively analyzed.None of them received adjuvant radiotherapy after mastectomy.The patients were divided into luminal A group,luminal B group,HER-2-overexpressing group,and triple-negative group according to the molecular subtypes of breast cancer determined by immunohistochemistry and fluorescence in situ hybridization.The patients in different groups were compared in terms of LR rate (LRR) and LR-free survival (LRFS),and the risk factors for LR were analyzed in combination with clinical and pathological features.The Kaplan-Meier method was adopted to calculate LRR and LRFS;the Logrank test was used for survival difference analysis and univariate prognostic analysis.The Cox proportional hazards model was used for multivariate prognostic analysis.Results The luminal A group,luminal B group,HER-2-overexpressing group,and triple-negative group accounted for 30.0％,48.6％,9.3％,and 12.1％,respectively,of all the patients.The follow-up rate was 97.1％ ;334 patients were followed up for at least 5 years,and 127 were followed up for at least 10 years.Univariate analysis showed that,compared with the luminal A group,the HER-2-overexpressing group and triple-negative group had significantly higher 5-year LRRs (19.0％ vs 5.3％,x2 =4.28,P =0.026; 14.9％ vs 5.3％,x2 =5.02,P =0.015) and significantly lower LRFSs (73.5％ vs 91.1％,x2=7.27,P=0.005;80.6％ vs 91.1％,x2=4.77,P=0.021).Multivariate analysis revealed that HER-2 overexpression,triple-negative phenotype,age of ≤ 35 years,and stage pT2 were poor prognostic factors for survival (LRR and LRFS) (x2 =2.29,2.08,18.22,and 6.86,P =0.020,0.016,0.001,and 0.005;x2 =1.90,1.41,8.58,and 3.94,P=0.006,0.025,0.002,and 0.039).The 10-year LRRs of patients with 0,1,and ≥2 of the above risk factors were 4.3％,14.1％,and 31.9％,respectively (x2 =28.03,P =0.000).Conclusions Molecular subtyping is helpful for individualized evaluation of LR risk in early breast cancer patients with 1-3 positive axillary lymph nodes (pN1).PMRT should be recommended for the patients with 2 or more risk factors for LR.

Objective:To study the effect of baicalein on the expression of glutamate receptor related protein in PC12 cells injured by Aβ 25-35. Methods:PC12 cells were divided into control group, model group, estradiol group and baicalein group with different concentrations. The survival condition of PC12 cells in each group were detected by thiazole blue (MTT). PC12 cells were divided into control group, model group, estradiol group and baicalein group. The control group and model group were cultured with DMEM medium, and the estradiol group was added with 1×10 -3 μmol/L estradiol DMEM medium, baicalein group was added with 1 μmol/L baicalein DMEM medium. After 2 hours of intervention, 20 μmol/L Aβ 25-35 was added to the model group, estradiol group and baicalein group with induced PC12 cell injury. After 22 hours of intervention, flow cytometry was used to detect the apoptosis of PC12 cells. The expression of estrogen receptor β (ER β), phosphorylated c-Jun N-terminal kinase (p-JNK/JNK) and ionic receptor N-methyl-D-aspartate receptor 1 (NMDAR1), glutamate receptor 2 (GluR2) and calcium/calmodulin dependent protein kinase Ⅱ (CaMK Ⅱ) were detected by Western blot. Results:Compared with model group, 1 μmol/L baicalein significantly increased the proliferation rate [(95.80±2.47)% vs. (64.34±3.84)%]. The apoptosis rate of PC12 cells[(7.83±0.67)% vs. (12.84±0.91)%] was significantly decreased in baicalein group ( P<0.01). The expression of NMDAR1 (0.582±0.012 vs. 0.352±0.012), GluR2(0.538±0.017 vs. 0.355±0.006), ER β (0.362±0.015 vs. 0.262±0.018) in baicalein group were significantly increased ( P<0.01), the expression of p-JNK/JNK (0.476±0.013 vs. 0.752±0.014) and CaMK Ⅱ(0.499±0.019 vs. 0.670±0.016) in baicalein group were significantly decreased ( P<0.01). Conclusions:Baicalein has a protective effect on PC12 cells injured by Aβ 25-35. Its mechanism may be related to the inhibition of p-JNK/JNK activity by activating ERβ and regulating the expression of glutamate receptor related protein.

Objective:To understand the status quo of emotional intelligence, emotional labor, and humanistic caring ability of medical staff, and to clarify their internal relationship.Methods:Convenience sampling was used to select 713 medical staff from a grade A tertiary hospital in Chengdu, Sichuan Province, China. Emotional Intelligence Scale, Humanistic Caring Scale, and Emotional Labor Scale were used to measure the emotional intelligence, humanistic caring ability, and emotional labor of medical staff. SPSS 22.0 software was used to establish a database for statistical description and analysis. Process 3.2 software was used to analyze the mediating effect.Results:In humanistic caring ability, the average score of comprehension dimension was the highest (75.62±8.20) and the average score of patience dimension was the lowest (58.53±5.01). In emotional labor, the average score of the deep action dimension was the highest (23.39±3.85) and the average score of the surface action dimension was the lowest (17.73±3.18). In emotional intelligence, the average score of self-emotion evaluation dimension was the highest (21.76±3.30) and the average score of other-emotion evaluation dimension was the lowest (20.07±3.71). Positive correlations were found between humanistic caring ability and emotional intelligence, between humanistic caring ability and emotional labor, and between emotional intelligence and emotional labor ( P<0.01). Humanistic caring ability had a partial mediating effect between emotional intelligence and emotional labor. Humanistic caring ability had direct and indirect effects on emotional labor, and the effect sizes were 0.279 and 0.029, respectively. Conclusion:Emotional intelligence has a direct positive predictive effect on emotional labor, humanistic caring ability as an intermediary variable indirectly and positively predicts emotional labor. Humanistic caring ability plays a partial mediating role between emotional intelligence and emotional labor. Attention should be paid to the emotional labor of medical staff, and the emotional labor of medical staff should be improved through targeted training on emotional intelligence and humanistic caring ability. These efforts will improve the current situation and establish a harmonious doctor-patient relationship.

ObjectiveTo investigate the effect of icariin （ICA）-mediated vitamin D system on peripheral blood dendritic cells （DCs） and helper T cells 17 （Th17）/regulatory T cells （Treg） balance in myocardial remodeling model of Dahl salt-sensitive rats. MethodFifty SPF Dahl salt-sensitive rats were divided into model group， vitamin D group （3×10^-5 mg·kg^-1·d^-1）， and high-， medium-， and low-dose ICA groups （120， 60， 30 mg·kg^-1·d^-1）， and 10 Dahl salt-resistant rats were used as normal group. The myocardial remodeling model was established by feeding rats with a high-salt diet containing 8% NaCl. After six weeks of modeling， the normal group and the model group were given an equal volume of ultrapure water by gavage， and other groups were continuously administrated for six weeks. Cardiac echocardiography， hematoxylin-eosin （HE） staining， and Masson staining were used to observe the pathological changes in cardiac structure and fibrosis. The levels of serum 25（OH）D₃， B-type N-terminal pro-brain natriuretic peptide （NT-ProBNP）， interleukin （IL）-17， transforming growth factor （TGF）-β₁， IL-12， and IL-10 were detected by enzyme-linked immunosorbent assay （ELISA）. The phenotype of peripheral blood DCs and the ratio of Th17/Treg cells of rats were detected by flow cytometry. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expressions of vitamin D receptor （VDR），1α-hydroxylase （CYP27B1）， and 24-hydroxylase （CYP24A1） in peripheral blood DCs of rats. ResultCompared with the control group， the rats in the model group had pathological changes such as disordered arrangement of myocardial cells and cytoplasmic hypertrophy and swelling. Myocardial collagen fibers proliferated significantly， and the arrangement of myocardial fibers was disordered. The levels of serum 25（OH）D₃ and IL-10 were significantly decreased， and the levels of serum IL-17， TGF-β₁， IL-6， IL-12， and NT-ProBNP were significantly increased （P<0.05）. The costimulatory molecules CD40， CD80， CD86， and MHC-Ⅱ were highly expressed in the peripheral blood DCs， and the expression of CD11 and CD11b was lower （P<0.05）. The proportion of Th17 cells in the peripheral blood was significantly increased， and the proportion of Treg cells was decreased. The ratio of Th17/Treg was increased （P<0.05）. The mRNA and protein expressions of CYP24A1 in peripheral blood DCs increased， and the mRNA and protein expressions of CYP27B1 and VDR decreased （P<0.05）. Compared with the model group， the arrangement of myocardial fibers in each drug administration group was relatively regular， and the swelling of myocardial cells was significantly reduced. The pathological morphology of myocardial tissue was improved to varying degrees. The pathological changes in myocardial tissue were improved and alleviated to varying degrees. The drug could reduce the serum levels of NT-ProBNP， IL-17， TGF-β₁， IL-6， and IL-12 and increase the level of serum 25（OH）D₃ and IL-10 （P<0.05）. The expression of costimulatory molecules CD40， CD80， CD86， and MHC-Ⅱ in the peripheral blood DCs of rats was decreased， and the expression of CD11 and CD11b molecules was increased （P<0.05）. The drug could reduce the proportion of Th17 cells in peripheral blood and the ratio of Th17/Treg cells and increase the proportion of Treg cells （P<0.05）. It could decrease the mRNA and protein expressions of CYP24A1 in peripheral blood DCs of rats and elevate the mRNA and protein expression of VDR and CYP27B1 （P<0.05）. ConclusionICA can regulate the phenotype of peripheral blood DCs and the ratio of Th17/Treg cells by regulating the vitamin D system and play a role in improving myocardial remodeling from the perspective of immune balance.

Sterol C24-methyltransferase (SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involved in the biosynthesis of 24-methyl sterols. Here, we report the cloning and characterization of a cDNA encoding a sterol C24-methyltransferase from().(GenBank access number KU885950) is a 1530 bp cDNA with a 1041 bp open reading frame predicted to encode a 346-amino acid, 38.62 kDa protein. The polypeptide encoded by thecDNA was expressed and purified as a recombinant protein from() and showed SMT activity. The expression ofwas highly up-regulated incell suspension cultures treated with methyl jasmonate (MeJA). Tissue expression pattern analysis showed higher expression in the phellem layer compared to the other four organs (leaf, stem, xylem and phloem), which is about ten times that of the lowest expression in leaf. The results are meaningful for the study of sterol biosynthesis ofand will further lay the foundations for the research in regulating both the content of other main compounds and growth and development of