A novel cell-wall polysaccharide derived from the stipe of Agaricus bisporus inhibits mouse melanoma proliferation and metastasis.

Malignant melanoma is an invasive and highly aggressive skin cancer that-if diagnosed-poses a serious threat to the patient's health and life. In this work, a novel purified cell-wall polysaccharide (termed Abwp) was obtained from the discarded stipe of Agaricus bisporus (A. bisporus) and characterized to be a novel homogeneous polysaccharide consisted of a ß-(1 â†’ 4)- glucosyl backbone with ß-(1 â†’ 2) and (1 â†’ 6)-d-glucosyl side-chains. The anti-melanoma effects of Abwp and its associated mechanisms in mice were then explored using in vitro and in vivo approaches. In vitro results showed that Abwp inhibited B16 melanoma cell proliferation and promoted their apoptosis in both time- and dose-dependent manners. In B16 cells induced with tumor necrosis factor (TNF-α), Abwp significantly decreased the protein expression of inflammatory-related signaling pathway (e.g., p38 MAPK and NF-κB) in time-, concentration-, and dose-dependent manners. Moreover, Abwp blocked nuclear entry of NF-κB-p65. In an in vivo mouse model featuring neoplasm transplantation with B16 melanoma cells, Abwp significantly inhibited the growth and proliferation of mouse melanoma. Hematoxylin staining showed that the invasion of melanoma cells into the lung tissue of the Abwp-treated group was significantly reduced. Immunohistochemical analysis showed that the expression of proliferation cell nuclear antigen (PCNA), N-cadherin, MMP-9, and Snail in the lung of mouse was significantly inhibited. Immunofluorescence showed that Abwp significantly interfered with the nuclear transcription of NF-κB-p65 in a dose-dependent manner. Collectively, these results showed that Abwp mediated p38 MAPK and NF-κB signaling pathways to inhibit the inflammatory response and malignant proliferation and metastasis of melanoma in mice.

Repairing and Analgesic Effects of Umbilical Cord Mesenchymal Stem Cell Transplantation in Mice with Spinal Cord Injury.

Transplantation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into spinal cord injury (SCI) may alleviate neuropathic pain and promote functional recovery. The underlying mechanism likely involves activation of glial cells and regulation of inflammatory factors but requires further validation. SCI was induced in 16 ICR mice using an SCI compression model, followed by injection of lentiviral vector-mediated green fluorescent protein- (GFP-) labeled hUC-MSCs 1 week later. Behavioral tests, histological evaluation, and inflammatory factor detection were performed in the treatment (SCI+hUC-MSCs) and model (SCI) groups. Histological evaluation revealed GFP expression in the spinal cord tissue of the treatment group, implying that the injected MSCs successfully migrated to the SCI. The Basso, Beattie, and Bresnahan (BBB) scores showed that motor function gradually recovered over time in both groups, but recovery speed was significantly higher in the treatment group than in the model group. The pain threshold in mice decreased after SCI but gradually increased over time owing to the self-repair function of the body. The corresponding pain threshold of the treatment group was significantly higher than that of the model group, indicating the therapeutic and analgesic effects of hUC-MSCs. Expression of IL-6 and TNF-α in the spinal cord tissue of the treated group decreased, whereas glial cell line-derived neurotrophic factor (GDNF) expression along with ED1 expression increased compared with those in the model group, suggesting that SCI activated ED1 inflammatory macrophages/microglia, which were subsequently reduced by hUC-MSC transplantation. hUC-MSCs are speculated to enhance the repair of the injured spinal cord tissue and exert an analgesic effect by reducing the secretion of inflammatory factors IL-6 and TNF-α and upregulating the expression of GDNF.

Prediction of kidney transplant outcome based on different DGF definitions in Chinese deceased donation.

BACKGROUND: Delayed graft function (DGF) is an important complication of kidney transplantation and can be diagnosed according to different definitions. DGF has been suggested to be associated with the long-term outcome of kidney transplantation surgery. However, the best DGF definition for predicting renal transplant outcomes in Chinese donations after cardiac death (DCDs) remains to be determined. METHOD: A total of 372 DCD kidney transplant recipients from June 2013 to July 2017 in the First Affiliated Hospital of Xi'an Jiaotong University were included in this retrospective study to compare 6 different DGF definitions. The relationships of the DGF definitions with transplant outcome were analyzed, including graft loss (GL) and death-censored graft loss (death-censored GL). Renal function indicators, including one-year estimated glomerular filtration rate (eGFR) and three-year eGFR, and were compared between different DGF groups. RESULTS: The incidence of DGF varied from 4.19 to 35.22% according to the different DGF diagnoses. All DGF definitions were significantly associated with three-year GL as well as death-censored GL. DGF based on requirement of hemodialysis within the first week had the best predictive value for GL (AUC 0.77), and DGF based on sCr variation during the first 3 days post-transplant had the best predictive value for three-year death-censored GL (AUC 0.79). Combination of the 48-h sCr reduction ratio and classical DGF can improve the AUC for GL (AUC 0.85) as well as the predictive accuracy for death-censored GL (83.3%). CONCLUSION: DGF was an independent risk factor for poor transplant outcome. The combination of need for hemodialysis within the first week and the 48-h serum creatinine reduction rate has a better predictive value for patient and poor graft outcome.

Clinical application of ultrasound-guided percutaneous microwave ablation for benign breast lesions: a prospective study.

BACKGROUND: Background: Benign breast lesions are the most common diseases in adult women, which have been treated with minimally invasive therapies in recent years. Little is known about the feasibility of Microwave ablation (MWA) for benign breast lesion treatment. The primary aim of this prospective study was to evaluate the safety and efficiency of MWA as a potential therapeutic option for benign breast lesions in a single-center cohort study. METHODS: Women with possibly benign breast lesions based on an ultrasound (US) assessment who were scheduled to undergo MWA between November 2014 to July 2018 were included in the study. The patients underwent conventional US to measure the size of the lesion, Doppler US to assess the vascularity of the lesion, elastography to evaluate the stiffness of the mass, core needle biopsy of suspicious lesions, contrast-enhanced US to help determine the treatment plan and eventually MWA of the lesion. Lesions were followed at one, three, six, twelve and eighteen months after treatment to with the same imaging modalities. RESULTS: A total of 314 women aged 17 to 69 years old (mean = 36.9 ± 9.9 years) with 725 benign breast lesions (mean of maximum diameter = 10.86 ± 5.40 mm) were included. The frequency of palpable mass, pain and nipple discharge significantly decreased after treatment. Complete ablation rate was 97.8%, immediately after ablation, which increased to 100% after supplementary ablation of the 15 cases with incomplete ablation. Blood flow classification and lesion's volume also showed a significant decrease, while both volume reduction ratio and disappearance rate significantly increased following treatment. The elasticity score of the lesions showed fluctuations across different follow-up intervals. None of the patients experienced major complications and the 1% who had mild symptoms were successfully treated. CONCLUSION: MWA treatment is shown to be safe and efficient and has the potential to be considered as an alternative first line treatment for benign breast lesions.

Effects of chronic unpredictable mild stress on ovarian reserve in female rats: Feasibility analysis of a rat model of premature ovarian failure.

Premature ovarian failure (POF) results from a number of disorders. The POF model is primarily based on chemotherapeutic injury, and hence is not suitable for assessing the effects of chronic stress on ovarian function. Therefore, improved animal models are required to analyze the effects of chronic stress on ovarian reserve. The feasibility of the chronic unpredictable mild stress (CUMS) method for establishing a model of POF was examined. The depressive behavior exhibited by rats was evaluated with the open field and sucrose preference tests. Vaginal smears were obtained for assessment of the estrous cycle. The ovarian reserve of the animals was evaluated using the estrous cycle, ovarian histology and serum levels of gonadotropin releasing hormone (GnRH), follicle­stimulating hormone (FSH), estradiol (E2), and anti­Müllerian hormone (AMH). Compared with the control group, body weight, time spent in the center, horizontal movement, vertical frequency, consumption of sucrose, sucrose preference, number of small follicles from the rats, and serum E2, AMH and GnRH levels were significantly decreased in the CUMS group (all P<0.05). However, the estrous cycle was prolonged significantly (P<0.05) and serum FSH levels were increased significantly (P<0.01). These results suggested that the CUMS model rats exhibited depression­like behaviors. CUMS may induce psychological stress and decrease ovarian reserve in female rats. Thus, the CUMS model may be used to assess the effects of chronic stress on female reproductive function.

Early Immunosuppressive Exposure of Enteric-Coated-Mycophenolate Sodium Plus Tacrolimus Associated with Acute Rejection in Expanded Criteria Donor Kidney Transplantation.

BACKGROUND: Immunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection (BPAR) after expanded criteria donor (ECD) kidney transplantation. The aim of this study was to investigate the relationships between early immunosuppressive exposure and the development of BPAR. METHODS: We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolimus (Tac), and prednisone. The levels of mycophenolic acid-area under the curve (MPA-AUC)0-12h and Tac C0were measured at the 1st week and the 1st month posttransplant, respectively. The correlation was assessed by multivariate logistic regression. RESULTS: The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC0-12h at the 1st week posttransplant was found in BPAR recipients (38.42 ± 8.37 vs. 50.64 ± 13.22, P < 0.01). In addition, the incidence of BPAR was significantly high (P < 0.05) when the MPA-AUC0-12hlevel was <30 mg·h-1·L-1 at the 1st week (15.0% vs. 44.4%) or the Tac C0was <4 ng/ml at the 1st month posttransplant (33.3% vs. 21.6%). Multivariable logistic regression analysis showed that the MPA-AUC0-12h at the 1st week (OR: 0.842, 95% CI: 0.784-0.903) and the Tac C0at the 1st month (OR: 0.904, 95% CI: 0.822-0.986) had significant inverse correlation with BPAR (P < 0.05). CONCLUSIONS: Low-level exposure of MPA and Tac C0in the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC0-12h <30 mg·h-1·L-1 and Tac C0 <4 ng/ml should be avoided in the first few weeks after transplantation.

Ultrasound-guided percutaneous microwave ablation for benign breast lesions: evaluated by contrast-enhanced ultrasound combined with magnetic resonance imaging.

BACKGROUND: Minimally invasive ablative techniques in the treatment of breast tumor has become popularly in recent years. METHODS: We analyzed gray-scale and contrast-enhanced ultrasound (CEUS) features of 205 microwave ablated breast benign lesions from 182 consecutive patients, compared with magnetic resonance imaging (MRI) and histopathology findings. The follow-up was implemented at 3, 6 and 12 months after the ablation treatment. RESULTS: Before the MWA, the mean of largest diameter and volume of the lesions were 14.41±6.54 and 3,224±961 mm3, respectively. However, those of the lesions respectively were 8.48±6.30 and 2,116±732 mm3 one year after the treatment. The longest diameter and the volume of the ablative lesions were gradually decreased 3, 6 or 12 months after the MWA. 44 (/205, 21.5%) ablative lesions were disappeared one year after the MWA. One hundred and forty-two (/205, 69.3%) ablative lesions presented a hypoechoic halo surrounding it on gray-scale US after the MWA. The success rate of the MWA treatment in the benign breast lesion was 87.32% and 82.93% evaluated by CEUS and enhanced MRI, respectively. During the ablation, no patient had serious complications, such as hemorrhage, serious pain and fat necrosis, etc. CONCLUSIONS: Microwave ablation was a safe and efficient method in the treatment of the benign breast tumors. CEUS and enhanced MRI could accurately assess whether the MWA treatment is effective.

[Evaluation of immune status of kidney transplant recipients by combined HLA-G5 and sCD30].

OBJECTIVE: to study the relationship between the expression of serum human leucocyte antigen-G5 (HLA-G5)/soluble CD30 (sCD30) and the function of renal graft in kidney transplant recipients and investigate the immune status of recipients with combined HLA-G5 and sCD30. METHODS: from January 2002 to November 2008, a total of 66 kidney transplant recipients in our centre were selected as subjects and divided into three groups: stable function of renal graft (n = 38), acute rejection (n = 15) and chronic rejection (n = 13). The expressions of serum HLA-G5 and sCD30 were detected. There were two different immune conditions with acute/chronic allograft rejection and normal renal graft in kidney transplant recipients as evaluated by combined HLA-G5 and sCD30. The sensitivity, specificity and critical value of the method were analyzed by the curve of receiver operating characteristic. RESULTS: the levels of HLA-G5 and sCD30 were significantly correlated with serum creatinine (r = -0.493, 0.691, both P < 0.01). Within the first year post-transplantation, the sensitivity was 78.6% and the specificity 85.7% when HLA-G5 critical value 82 microg/L and sCD30 critical value 12.2 microg/L. After one year post-transplantation: the sensitivity was 92.3% and the specificity 84.6% when HLA-G5 critical value 141 microg/L and sCD30 critical value 10.3 microg/L. CONCLUSION: the immune state of recipients are evaluated by combine HLA-G5 and sCD30 which may be a simple and valid method.

[The effects of ICA on the cell-immunosuppressive and bone-marrow-suppressive mice after chemotherapy].

AIM: to observe the effect of ICA on the immunosuppressive and bone-marrow-suppressive mice after chemotherapy, and explore the machanism of hematopoietic and immunologic function in mice accentuated by ICA. METHODS: mice were injected Cy intraperitoneally except control group , then randomly divided into mode group(saline), positive (Shenqi, 1 mL/d)group, ICA groups(150, 80, 40 mg/kg.d). All mice were treated respectively for 10 successive days. The pathological changes of thymus were observed by HE staining. Killing activity of peritoneal macrophage were measured by LDH kits and its production of TNF-α and IL-12 was measured by ELISA kits. Population of white blood cells (WBC), red blood cell (RBC) and platelet (PLT) in the peripheral blood were detected with automated blood cell counter (ABCC). Hemogram of peripheral blood and bone marrow morphology were observed under the microscope. RESULTS: ICA could protect the thymus and bone marrow from damage. The proliferation of lymphocyte and killing activity of macrophage cells in ICA treatment groups was all enhanced, moreover, the population of WBC, RBC and PLT were increased significantly. CONCLUSION: ICA can improve the state of immunosuppressive and myelosuppressive mice caused by Cy thus could alleviate the side effect of chemotherapy effectively.

[Recombinant adenovirus-mediated human cytosolic glutathione peroxidase gene transfection protects vascular endothelial cells from oxidative damage].

OBJECTIVE: To study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage. METHODS: pGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2). RESULTS: The expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage. CONCLUSION: hCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.

[Protection of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by recombinant adenovirus containing hCGPx gene].

AIM: To study the protective effect of human renal tubular epithelial cells (HK-2 cells) from hypoxia-reoxygenation damage by transfection of recombinant adenovirus-mediated human cytosolic glutathione peroxidase(hCGPx) gene. METHODS: Recombinant pGEM-T vector containing hCGPx cDNA and pACCMV-pLpA adenovirus shuttle plasmid was constructed. Then the shuttle plasmid pACCMV-hCGPx and pJM17 were co-transfected into 293 cells and recombinant adenovirus AdCMV-hCGPx was obtained. Cultured HK-2 cells were transfected with AdCMV-hCGPx or vacant recombinant adenovirus (control). The expression ratio of transfected hCGPx gene were studied. Cell viability, the percentage of apoptosis and death were evaluated after hypoxia-reoxygenation damage. RESULTS: The expression ratio of hCGPx gene was higher in the AdCMV-hCGPx transfected cells than that in the control group (P<0.01). After hypoxia-reoxygenation damage, the viability of hCGPx gene transfected cells was significantly higher than that of control and the percentage of apoptosis and death of hCGPx transfected cells was significantly lower than that of control. CONCLUSION: The transfection of hCGPx mediated by recombinant adenovirus could protect renal tubular epithelial cells from hypoxia-reoxygenation damage in vitro.