Objective:To investigate the expression profile of IL-36 family members in patients with coronary atherosclerotic heart disease (CAHD) and to assess the regulatory effects of exogenous IL-36 on CD8 + T cell function in CAHD patients. Methods:Twenty controls and 82 CAHD patients including 31 with stable angina pectoris (SAP), 27 with unstable angina pectoris (UAP) and 24 with acute myocardial infarction (AMI) were enrolled in this study. Anti-coagulant peripheral blood samples were collected. Plasma and peripheral blood mononuclear cells were isolated. The levels of IL-36α, IL-36β, IL-36γ and IL-36 receptor antagonist (IL-36RA) in plasma were measured by ELISA. CD8 + T cells were enriched. The expression of IL-36 receptor subunits at mRNA level was semi-quantified by real time PCR. Flow cytometry was used to detect the expression of programmed death-1 (PD-1), cytotoxic T lymphocytes associated protein-4(CTLA-4) and lymphocyte-activation gene-3 (LAG-3) in CD8 + T cells. Levels of periforin, granzyme B, granulysin, IFN-γ and TNF-α in the culture supernatants of CD8 + T cells were measured by ELISA. Purified CD8 + T cells from controls and AMI patients were stimulated with recombinant human IL-36RA. Changes in the expression of immune checkpoint molecules and the secretion of cytotoxic molecules and cytokines after IL-36RA stimulation were analyzed. One-way analysis of variance or paired t-test was used for statistical analysis. Results:There were no significant differences in plasma IL-36α, IL-36β or IL-36γ level between the control, SAP, UAP and AMI groups ( P>0.05). Plasma IL-36RA level was significantly down-regulated in the AMI group as compared with that in the control, SAP and UAP groups[(1 159.57±297.83) pg/ml vs (1 773.47±754.29) pg/ml, (1 600.12±740.48) pg/ml and (1 578.72±720.42) pg/ml; P<0.05]. The expression of IL-1 receptor 6 (IL-1R6) and IL-1 receptor accessory protein (IL-1RAcP) at mRNA level, the expression of PD-1 and CTLA-4, and the secretion of IFN-γ and TNF-α by CD8 + T cells showed no significant differences between the four groups ( P>0.05). Periforin, granzyme B and granulysin levels secreted by CD8 + T cells of the AMI group were significantly higherthan those of the control, SAP and UAP groups ( P<0.05). In the control group, recombinant human IL-36RA stimulation did not affect the expression of immune checkpoint molecule or the secretion of cytotoxic molecules and cytokines by CD8 + T cells ( P>0.05). In the AMI group, the percentage of PD-1 + CD8 + T cells increased after recombinant human IL-36RA stimulation ( P=0.033), but no significant change in the percentage of CTLA-4 + CD8 + T cells was observed ( P=0.288). Moreover, recombinant human IL-36RA stimulation suppressed the CD8 + T cells of AMI patients to secrete periforin, granzyme B and granulysin ( P<0.05), but not affect the secretion of IFN-γ and TNF-α ( P>0.05). Conclusions:The reduced IL-36RA level in AMI patients might induce the enhancement of CD8 + T cell activity by promoting CD8 + T cells to secrete cytotoxic molecules, which was involved in the immunopathogenesis of AMI.

Objective:To observe any effect of alternate side whole body vibration on the muscle strength, balance and walking ability of persons with sarcopenia.Methods:Forty elderly persons with sarcopenia were randomly divided into an experimental group and a control group, each of 20. Both groups received conventional balance and lower-limb muscle strength training, but the experimental group also received 9-14Hz whole body vibration treatment with an amplitude of 2-3mm, 5 one-minute groups a day, 5 times a week. Before and after 8 weeks of treatment, both groups′ lower limb strength, 6m step speed, exercise ellipse area, exercise track length, and Up and Go test (TUGT) time were measured.Results:In the experimental group the average bilateral iliopsoas muscle strength and all the other measurements had improved significantly. In the control group, the exercise ellipse area with the eyes closed and the exercise track length with the eyes open and all the other measurements had improved significantly compared with before the training. But all of the experimental group′s outcomes were, on average, significantly better than those of the control group.Conclusion:Low frequency vibration training can better improve the muscle strength, balance and walking ability of elderly persons with sarcopenia. It can be used for prevention as well as treatment.

Background The mechanisms of silicon dioxide (SiO₂)-induced inflammation and cell injury in pulmonary macrophages are not fully characterized. Objective To investigate the potential roles of inhibition of toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling in inflammation and macrophage polarization in mouse Raw264.7 cells in response to SiO₂ stimulation. Methods Sixteen 6- to 8-week-old C57BL/6 mice, half male and half female, were intratracheally instilled with 50 µL of SiO₂ (50 mg·mL⁻¹ in saline) or normal saline via oropharyngeal route, and the lungs of mice were harvested at 14 d and 28 d post the first challenge of SiO₂. HE staining of mouse lung was used for histopathological analysis. The expressions of TLR4 signaling-related proteins were detected by Western blotting (WB) and immunofluorescent (IF) assay, including TLR4, myeloid differentiation factor 88 (Myd88), and TNF receptor associated factor 6 (TRAF6). Raw264.7 cells were stimulated with SiO₂ (100 μg·cm²) for 12 h in absence or presence of TLR4 inhibitor M62812 for 13 h before the culture supernatants and cell lysates were harvested for analysis. The expressions of key components of TLR4 signaling cascade including TLR4, Myd88, and phosphorylated nuclear factor-kappa B P65 (P-NF-κB P65), P-1NF-kappa-B inhibitor α (P-1κbα), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6), M1 phenotype markers inducible nitric oxide synthase (iNOS) and cluster of differentiation 86 (CD86), as well as M2 phenotype arginase-1 (Arg-1) were accessed by WB and IF. The expressions of inflammation factors IL-6 and TNF-α in supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Results After SiO₂ intratracheal instillation for 14 d, the HE staining results showed obvious fibrotic nodules in the lung tissues of mice. The results of WB analysis revealed more abundant TLR4, Myd88, and TRAF6 in the silicosis mouse lung samples than in the controls. The results of IF assay showed an increased abundance of TLR4 and Myd88 proteins in the lung samples of silicosis mice at 14 d post the silica challenge, compared to the controls, indicating TLR4 signaling activation. As seen in the in vitro experiment, significant upregulations after the exposure to 100 μg·cm² SiO₂ were observed in TLR4 and P-1κbα at 6, 12, and 24 h (P<0.05); Myd88 at 12 and 24 h (P <0.05); and P-NF-κB P65 at 12 h (P<0.05). The inhibitor significantly suppressed the expressions of TLR4, Myd88, TRAF6, P-NF-κB P65, TNF-α, and IL-6 in Raw264.7 cells. In addition, the SiO₂-induced M1 phenotype marker iNOS was significantly suppressed, but the M2 phenotype marker Arg-1 was increased in the Raw264.7 cells. Conclusion The inhibition of TLR4/NF-κB signaling could result in a reduction of the inflammation response and the transition of M1 toward M2 phenotypes of macrophages in response to SiO₂ challenge.

Objective To investigate whether there are differences of the characteristics of the contrast‐enhanced ultrasound among breast cancer vary in size .Methods The contrast‐enhanced ultrasound perfusion mode were retrospectively analyzed in 113 cases of breast cancer patients with 119 lesions .They were divided into two groups according to the maximum diameter of the lesion based on ultrasound measured ,one group (≤2 0. cm ) 47 cases ,the other group (> 2 0. cm ) 72 cases .The characteristics and mode of contrast‐enhanced ultrasound of the lesion were analyzed ,including whether is greater than the two‐dimensional ultrasound in enhanced range ,enhanced strength ,the distribution of the contrast agent ,contrast agent perfusion sequence in lesions ,contrast mode ,and the existence of perforator vessels and perfusion defects .Results Enhanced strength ,the existence of perforator vessels and perfusion defects associated with the size of the lesions .The breast cancer lesions more than 20 cm were apt to higher enhance strength ,the presence of perfusion defects ,perforator vessels ( P < 0 0.5) .However ,there were something that had nothing to do with the size of the lesions including whether or not contrast‐enhanced ultrasound range was greater than the two‐dimensional ultrasound range ,contrast agent distribution characteristics ,contrast agent perfusion sequence in lesions ,and differences in contrast patterns ( P >0 0.5) .Regardless of breast lesions size ,the enhanced range tended to larger than two‐dimensional ultrasound range ,and contrast‐enhanced ultrasound present inhomogeneous ,centrality and integrity filling and perfusion mode of quickly rising and falling .Conclusions Within breast tumor angiogenesis is gradual .With the increase of tumor volume ,more significant vascular heterogeneity ,breast cancer lesion are more prone to uneven high perfusion ,accompanied by perfusion defects and perforating vessels ,but the main sign of differentiating benign and malignant showed consistency between different sized lumps in breast cancer ,which included whether or not contrast mode and enhanced range greater than two‐dimensional ultrasonic range .

Objective:To investigate the role of alloreactive T cell in clonal deletion and regulatory T cells ( Treg) in transplant tolerance.Methods:F1 mice were bred by crossing female BALB/c mice and male C57BL/6 mice.Within 24 h,newborn C57BL/6 mice were inoculated with F1 spleen cells via the orbital branch of the anterior facial vein.Six weeks later,the mice were subjected to F1 skin grafting to evaluate their tolerance.Proliferation,flow cytometry and adoptive transfer assay were used to analyze clonal deletion of alloreactive T cells and the expression of CD4+Foxp3+T cells in neonatal treated mice.Results: Newborn C57BL/6 mice injected with F1 splenic cells could induce transplantation tolerance,the level of tolerance was associated with the dose of splenic cells.3×107 splenic cells from F1 mice could induce long-term skin graft acceptance in C57BL/6 mice ,1×107splenic cells significantly prolonged the survival of F1 skin grafts,but the grafts completely rejected within 50 days.The mixed lymphocyte reaction ( MLR) experiment in vivo showed that alloreactive T cell in long-term tolerant mice was deleted completely,but a certain amount of reactive T cells existed in the low-dose group mice.Flow cytometry ( FCM) analysis showed that the expression of CD4+Foxp3+T cell in the high-dose group and low-dose group mice had no obvious difference compared with the naive mice.When alloreactive T-cells were injected into tolerant mice,the skin graft rejection was observed,and Treg cells upregulated in graft-rejected mice.Conclusion:The degree of transplantation tolerance depended on the clonal deletion of alloreactive T cells,instead of on the expression of CD4+Foxp3+Treg cells.CD4+Foxp3+regulatory T cells upregulated in graft rejected mice,which may be served as a negative feedback mechanism to control the intensity of rejection.

Objective To study the ultrasonic imaging of breast cancer with or without filling defect with contrast-enhanced ultrasound and clinical pathologic correlation.Methods The suspicious breast lesions underwent contrast-enhanced ultrasound examination, the imaging of 1 1 9 breast cancer with pathological results were retrospectively analyzed,and the contrast enhanced ultrasound effect,distribution of contrast agent,sequence of contrast agent into the tumor,contrast enhanced ultrasound mode,perforator vessel,filling defect and flow classification were observed.Then clinical and pathological data were collected including the tumor size,clinical stage,pathological grade and the situation of micro calcification.The perfusion defect as the grouping factor,there were 37 cases of perfusion defect and 82 cases without perfusion defect.A mono-pactor and multi factor analysis was performed by the logistic regression models to the relationship between perfusion defect and contrast-enhanced ultrasonography,clinical pathological parameters.Results Both mono factor and multifactor analysis displayed that whether to display the filling defect in contrast enhanced ultrasound was associated with ultrasound contrast distribution,the sequence of contrast agent into the tumor and histological grade(χ2=19.5050,21.1210,10.3640,OR =6.977,0.261, 2.509,all P <0.05).Multivariate analysis showed that tumor size and whether to display the filling defect in contrast enhanced ultrasound were close(OR =1.692,P <0.05).Clinical stage,whether the blood flow signal penetrates was meaningful in univariate analysis but excluded in multivariate analysis.Both mono factor and multifactor analysis displayed that contrast enhanced ultrasound mode,contrast enhanced ultrasound effect,flow classification and pathological micro calcification were not associated with whether to display the filling defect.Conclusions Whether to display the filling defect and the related factors of contrast-enhanced ultrasound in diagnosis of benign and malignant were not completely consistent.Breast cancers with filling defects of contrast-enhanced ultrasound tend to present large masses,contrast agent distribution of peripheral enhancement,centripetal filling of ultrasound contrast agents and the higer histological grading.

Objective To explore the application value of contrast-enhanced ultrasound(CEUS) in evaluating the effect of neoadjuvant chemotherapy(NAC) on breast cancer.Methods Image features of 80 breast cancer patients with neoadjuvant chemotherapy were observed and analyzed by CEUS.In the process,the changes of primary foci before and after neoadjuvant chemotherapy and the changes of qualitative and quantitative indexes of blood perfusion were evaluated.The correlation between qualitative change and pathological response was also studied.Results The change of focus size measured by CEUS was higher than that of conventional ultrasound before and after neoadjuvant chemotherapy(P ＜0.05).However,there was no significant difference between the two measurements before NAC(P ＞ 0.05).All the qualitative observation indexes of the ultrasound contrast after NAC revealed obviously benign conversion(P ＜0.05).Among them,the pathological response was closely related to the changes of the enhancement degree,the distribution of the contrast agent,the order of the contrast and the contrast mode.After NAC,the perfusion parameters of the lesions showed slowly crawl and slowly retreat,low enhancement and the diminished total amount of microcirculation perfusion.Conclusions Before and after neoadjuvant chemotherapy,the changes of the lesion size deteted by CEUS were more close to the pathological evaluation than those detected by conventional ultrasound.The benign outcome of qualitative and quantitative indexes of the CEUS was more obvious than the changes of the lesion size,which had practical value for guiding the neoadjuvant chemotherapy for breast cancer,especially the CEUS indexes related to pathological assessment.

Purpose Meta-analysis was performed to compare elastosonography and routine ultrasonography in the diagnosis of thyroid nodules. Materials and Methods Extensive literature search of CNKI, Wanfang data, OVID, the Cochrane Library, PubMed and Embase from 1980 to 2013 was performed using elastosonography, ultrasonography and thyroid nodules as key words. Clinical trials utilizing elastosonograpy and routine ultrasonography were selected. The quality and heterogeneity were evaluated. Specificity and specificity were summarized to generate receiver operating characteristic curves. Results Fourteen studies were selected including 10 literature in Chinese and 4 in English. There was significant heterogeneity in the sensitivity. The sensitivity for routine ultrasound, elastosonography and two techniques combined was 0.82 (95%CI 0.79-0.85), 0.61 (95%CI 0.57-0.64) and 0.93 (95%CI 0.90-0.95), respectively;with specificity of 0.70 (95%CI 0.68-0.72), 0.84 (95%CI 0.82-0.86) and 0.69 (95%CI 0.67-0.72). The summery area under curve was 0.8549, 0.8925 and 0.9614, respectively. Conclusion Evaluation of thyroid nodules using elastosonography is more sensitive and specific than routine ultrasonography. Combining elastosonography and routine ultrasonography provides better diagnostic accuracy.

Objectives:To determine the expressions of decoy receptors (DcR1 and DcR2) of TRAIL in carcinoma of endometrium. Methods:The expressions of DcR1 and DcR2 in endometrium tissues from 13 carcinoma of endometrium and 7 normal endometrium were detected by immunohistochemical staining.Results: The expressions of DcR1 and DcR2 in carcinoma of endometrium were much lower than in normal endometrium. Conclusions:The decreasing of DcR1 and DcR2 in carcinoma of endometrium may be concerned with its pathogenesis, which may be related to the prevention of endometrium from carcinomatous change.

Objective To review the clinical-pathological features, the tendency of incidence over 20 years, the predisposing factors, and the differences between the cases of cutaneous malignant melanoma from two hospitals. Methods A collection of 305 cases diagnosed as cutaneous malignant melanoma, among which 185 cases had complete clinical-pathological data, during 1981-2000 was analyzed. Results Acral malignant melanoma accounted for 63.3%, and the cases associated with congenital small nevi at the primary site accounted for 15.8% of 305 patients. During the period 1981-1990 and 1991-2000, cutaneous malignant melanoma constituted 0.053% and 0.094%, respectively, of all diagnoses with pathological sections, with an growth rate of 3.9% yearly. There was a tendency of the increased lesions located on face and neck, and decreased lesions on acra, over 20 years. Conclusion A rise of diagnosis of cutaneous malignant melanoma has been noticed from two hospitals over 20 years. Acra, especially planta, is the predominant anatomical site of cutaneous malignant melanoma.