Effect of CXCR3/HO-1 genes modified bone marrow mesenchymal stem cells on small bowel transplant rejection.

AIM: To investigate whether bone marrow mesenchymal stem cells (BMMSCs) modified with the HO-1 and CXCR3 genes can augment the inhibitory effect of BMMSCs on small bowel transplant rejection. METHODS: Lewis rat BMMSCs were cultured in vitro. Third-passage BMMSCs were transduced with the CXCR3/HO-1 genes or the HO-1 gene alone. The rats were divided into six groups and rats in the experimental group were pretreated with BMMSCs 7 d prior to small bowel transplant. Six time points (instant, 1 d, 3 d, 7 d, 10 d, and 14 d) (n = 6) were chosen for each group. Hematoxylin-eosin staining was used to observe pathologic rejection, while immunohistochemistry and Western blot were used to detect protein expression. Flow cytometry was used to detect T lymphocytes and enzyme linked immunosorbent assay was used to detect cytokines. RESULTS: The median survival time of BMMSCs from the CXCR3/HO-1 modified group (53 d) was significantly longer than that of the HO-1 modified BMMSCs group (39 d), the BMMSCs group (26 d), and the NS group (control group) (16 d) (P < 0.05). Compared with BMMSCs from the HO-1 modified BMMSCs, BMMSCs, and NS groups, rejection of the small bowel in the CXCR3/HO-1 modified group was significantly reduced, while the weight of transplant recipients was also significantly decreased (P < 0.05). Furthermore, IL-2, IL-6, IL-17, IFN-γ, and TNF-α levels were significantly decreased and the levels of IL-10 and TGF-ß were significantly increased (P < 0.05). CONCLUSION: BMMSCs modified with the CXCR3 and HO-1 genes can abrogate the rejection of transplanted small bowel more effectively and significantly increase the survival time of rats that receive a small bowel transplant.

Effects of heme oxygenase-1-modified bone marrow mesenchymal stem cells on microcirculation and energy metabolism following liver transplantation.

AIM: To investigate the effects of heme oxygenase-1 (HO-1)-modified bone marrow mesenchymal stem cells (BMMSCs) on the microcirculation and energy metabolism of hepatic sinusoids following reduced-size liver transplantation (RLT) in a rat model. METHODS: BMMSCs were isolated and cultured in vitro using an adherent method, and then transduced with HO-1-bearing recombinant adenovirus to construct HO-1/BMMSCs. A rat acute rejection model following 50% RLT was established using a two-cuff technique. Recipients were divided into three groups based on the treatment received: normal saline (NS), BMMSCs and HO-1/BMMSCs. Liver function was examined at six time points. The levels of endothelin-1 (ET-1), endothelial nitric-oxide synthase (eNOS), inducible nitric-oxide synthase (iNOS), nitric oxide (NO), and hyaluronic acid (HA) were detected using an enzyme-linked immunosorbent assay. The portal vein pressure (PVP) was detected by Power Lab ML880. The expressions of ET-1, iNOS, eNOS, and von Willebrand factor (vWF) protein in the transplanted liver were detected using immunohistochemistry and Western blotting. ATPase in the transplanted liver was detected by chemical colorimetry, and the ultrastructural changes were observed under a transmission electron microscope. RESULTS: HO-1/BMMSCs could alleviate the pathological changes and rejection activity index of the transplanted liver, and improve the liver function of rats following 50% RLT, with statistically significant differences compared with those of the NS group and BMMSCs group (P < 0.05). In term of the microcirculation of hepatic sinusoids: The PVP on POD7 decreased significantly in the HO-1/BMMSCs and BMMSCs groups compared with that of the NS group (P < 0.01); HO-1/BMMSCs could inhibit the expressions of ET-1 and iNOS, increase the expressions of eNOS and inhibit amounts of NO production, and maintain the equilibrium of ET-1/NO (P < 0.05); and HO-1/BMMSCs increased the expression of vWF in hepatic sinusoidal endothelial cells (SECs), and promoted the degradation of HA, compared with those of the NS group and BMMSCs group (P < 0.05). In term of the energy metabolism of the transplanted liver, HO-1/BMMSCs repaired the damaged mitochondria, and improved the activity of mitochondrial aspartate aminotransferase (ASTm) and ATPase, compared with the other two groups (P <0.05). CONCLUSION: HO-1/BMMSCs can improve the microcirculation of hepatic sinusoids significantly, and recover the energy metabolism of damaged hepatocytes in rats following RLT, thus protecting the transplanted liver.

Biological effects of bone marrow mesenchymal stem cells on hepatitis B virus in vitro.

The aim of the present study was to explore the effects of co­culturing bone marrow­derived mesenchymal stem cells (BM-MSCs) cultured with hepatitis B virus (HBV)­infected lymphocytes in vitro. BM­MSCs and lymphocytes from Brown Norway rats were obtained from the bone marrow and spleen, respectively. Rats were divided into the following five experimental groups: Group 1, splenic lymphocytes (SLCs); group 2, HepG2.2.15 cells; group 3, BM­MSCs + HepG2.2.15 cells; group 4, SLCs + HepG2.2.15 cells; and group 5, SLCs + BM­MSCs + HepG2.2.15 cells. The viability of lymphocytes and HepG2.2.15 cells was assessed using the MTT assay at 24, 48 and 72 h, respectively. Levels of supernatant HBV DNA and intracellular HBV covalently closed circular DNA (cccDNA) were measured using quantitative polymerase chain reaction. Supernatant cytokine levels were measured by enzyme­linked immunosorbent assay (ELISA). T cell subsets were quantified by flow cytometry using fluorescence­labeled antibodies. In addition, the HBV genome sequence was analyzed by direct gene sequencing. Levels of HBV DNA and cccDNA in group 5 were lower when compared with those in group 3 or group 4, with a significant difference observed at 48 h. The secretion of interferon­Î³ was negatively correlated with the level of HBV DNA, whereas secretion of interleukin (IL)­10 and IL­22 were positively correlated with the level of HBV DNA. Flow cytometry demonstrated that the percentage of CD3+CD8+ T cells was positively correlated with the levels of HBV DNA, and the CD3+CD4+/CD3+CD8+ ratio was negatively correlated with the level of HBV DNA. Almost no mutations in the HBV DNA sequence were detected in HepG2.2.15 cells co­cultured with BM­MSCs, SLCs, or in the two types of cells combined. BM­MSCs inhibited the expression of HBV DNA and enhanced the clearance of HBV, which may have been mediated by the regulation of the Tc1/Tc2 cell balance and the mode of cytokine secretion to modulate cytokine expression.

Effect of heme oxygenase-1 transduced bone marrow mesenchymal stem cells on damaged intestinal epithelial cells in vitro.

In this study, we explored the effects of mesenchymal stem cells (MSCs) from bone marrow overexpressing heme oxygenase-1 (HO-1) on the damaged human intestinal epithelial barrier in vitro. Rat MSCs were isolated from bone marrow and transduced with rat HO-1 recombinant adenovirus (HO-MSCs) for stable expression of HO-1. Colorectal adenocarinoma 2 (Caco2) cells were treated with tumor necrosis factor-α (TNF-α) to establish a damaged colon epithelial model. Damaged Caco2 were cocultured with MSCs, Ad-MSCs, Ad-HO + MSCs or HO-MSCs. mRNA and protein expression of Zona occludens-1 (ZO-1) and human HO-1 and the release of cytokines were measured. ZO-1 and human HO-1 in Caco2 were significantly decreased after treatment with TNF-α; and this effect was reduced when coculture with MSCs from bone marrow. Expression of ZO-1 was not significantly affected by Caco2 treatment with TNF-α, Ad-HO, and MSCs. In contrast, ZO-1 and human HO-1 increased significantly when the damaged Caco2 was treated with HO-MSCs. HO-MSCs showed the strongest effect on the expression of ZO-1 in colon epithelial cells. Coculture with HO-MSCs showed the most significant effects on reducing the expression of IL-2, IL-6, IFN-γ and increasing the expression of IL-10. HO-MSCs protected the intestinal epithelial barrier, in which endogenous HO-1 was involved. HO-MSCs play an important role in the repair process by reducing the release of inflammatory cytokines and increasing the release of anti-inflammatory factors. These results suggested that HO-MSCs from bone marrow were more effective in repairing the damaged intestinal epithelial barrier, and the effectiveness of MSCs was improved by HO-1 gene transduction, which provides favorable support for the application of stem cell therapy in the intestinal diseases.

Immunomodulatory effects of bone marrow mesenchymal stem cells overexpressing heme oxygenase-1: Protective effects on acute rejection following reduced-size liver transplantation in a rat model.

Here we explore the T-lymphocyte suppressive and immunomodulatory effects of bone marrow mesenchymal stem cells (BMMSCs) overexpressing heme oxygenase-1 (HO-1) on acute rejection following reduced-size liver transplantation (RLT) in a rat model. The proliferation activity, cell cycle progression, secretion of proinflammatory cytokines, expression of CD25 and CD71 in lymphocytes, and activity of NK cells were found to be significantly lowered, and the proportion of regulatory T cells (Tregs) was found to be increased relative to BMMSCs when Adv-HO-1/BMMSCs were co-cultured with Con A ex vivo; secretion of anti-inflammatory cytokines was significantly higher. When treated with saline, BMMSCs or Adv-HO-1/BMMSCs, post-transplantation rats receiving Adv-HO-1/BMMSCs showed better median survival time, lower rejection activity index, higher anti-inflammatory cytokine levels, lower proinflammatory cytokine levels, more peripheral Tregs, and lower natural killer cell viability. These results suggest that HO-1 enhanced and prolonged the effects of BMMSCs on acute rejection following RLT, with immunomodulatory effects in which adaptive and innate immunity, as well as paracrine signaling, may play important roles.

Mesenchymal stem cells improve the outcomes of liver recipients via regulating CD4+ T helper cytokines in rats.

BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) exert immunosuppressive activities in transplantation. This study aimed to determine whether BMMSCs reduce acute rejection and improve outcomes of liver transplantation in rats. METHODS: Orthotopic liver transplantation from Lewis to Brown Norway rats was performed, which was followed by the infusion of BMMSCs through the penile superficial dorsal vein. Normal saline infusion was used as a control. Animals were sacrificed at 0, 24, 72, or 168 hours after BMMSCs infusion. Liver grafts, and recipient serum and spleen tissues were obtained. Histopathology, apoptosis, serum liver enzymes, serum cytokines, and circulating regulatory T (Treg), Th1, Th2 and Th17 cells were assessed at each time point. RESULTS: BMMSCs significantly attenuated acute rejection and improved the survival rate of allogeneic liver transplantation recipients. Liver enzymes and liver apoptosis were significantly alleviated. The levels of the Th1/Th2 ratio-associated cytokines such as IL-2 and IFN-gamma were significantly reduced and IL-10 was significantly increased. The levels of the Th17/Tregs axis-associated cytokines such as IL-6, IL-17, IL-23, and TNF-alpha were significantly reduced, whereas TGF-beta concentration was significantly increased. Moreover, flow cytometry analysis showed that the infusion of BMMSCs significantly increased Th2 and Treg cells and decreased Th1 and Th17 cells. CONCLUSION: BMMSCs had immunomodulatory effects, attenuated acute rejection and improved outcomes of allogeneic liver transplantation in rats by regulating the levels of cytokines associated with Th1/Th2 and Th17/Treg ratios.

Improvement of Liver Transplantation Outcome by Heme Oxygenase-1-Transduced Bone Marrow Mesenchymal Stem Cells in Rats.

Bone marrow mesenchymal stem cells (BMMSCs) exert immunosuppressive activity in transplantation, and heme oxygenase-1 (HO-1) enhances their immunomodulatory effects. The aim of this study was to determine whether HO-1-transduced BMMSCs (HO-1/MSCs) improve rat liver transplantation (LTx) outcomes. Orthotopic LTx rejection models were treated with HO-1/MSCs, BMMSCs, HO-1, or normal saline, respectively. Our results showed a significant improvement in survival rates in the HO-1/BMMSCs group compared to the control groups. At all time points, liver function marker levels in the HO-1/MSCs group were significantly lower than in the other three groups; on POD 1, 7, and 14, the degree of rejection and apoptotic cells was significantly less in the HO-1/MSCs group than in the other three groups. Interleukin- (IL-) 10 and transforming growth factor-ß levels were significantly increased, while IL-2, IL-6, IL-17, IL-23, tumor necrosis factor-α, and interferon-γ levels were significantly decreased in the HO-1/MSCs group when compared to the other groups. Splenocyte Tregs were significantly increased by HO-1/MSCs compared with controls on POD 3, 5, 7, 10, 14, and 28. Summarily, we provide evidence that HO-1/MSCs improved allogeneic LTx outcomes by attenuating inflammatory responses and acute cellular rejection, as well as enhanced immunomodulatory effects compared with BMMSCs.

Influence of thyroid dysfunction on male sexual and reproductive functions and its mechanisms / 中华男科学杂志

Thyroid dysfunction may affect the body growth metabolism and the synthesis and secretion of sex steroids. A large number of clinical and basic studies suggest that thyroid hormone abnormality can cause sexual and reproductive problems. Patients with hyperthyroidism or hypothyroidism may develop not only premature ejaculation but also erectile dysfunction. The thyroid hormone level also influences Leydig cells, Sertoli cells, and spermatogenesis. This review focuses on the association of thyroid dysfunction with male premature ejaculation, erectile dysfunction, and infertility and its mechanisms.

Ag(x)@WO3 core-shell nanostructure for LSP enhanced chemical sensors.

Exceptional properties of graphene have triggered intensive research on other 2D materials. Surface plasmon is another subject being actively explored for many applications. Herein we report a new class of core-shell nanostructure in which the shell is made of a 2D material for effective plasmonic propagation. We have designed a much enhanced chemical sensor made of plasmonic Ag(x)@(2D-WO3) that combines above advantages. Specifically, the sensor response increases from 38 for Ag(x)-WO3 mixture to 217 for the Ag(x)@(2D-WO3) core-shell structure; response and recovery time are shortened considerably to 2 and 5 seconds; and optimum sensor working temperature is lowered from 370°C to 340°C. Light irradiation is found to increase the Ag(x)@(2D-WO3) sensor response, particularly at blue wavelength where it resonates with the absorption of Ag nanoparticles. Raman scattering shows significantly enhanced intensity for both the 2D-WO3 shell and surface adsorbates. Both the resonance sensor enhancement and the Raman suggest that the improved sensor performance is due to nanoplasmonic mechanism. It is demonstrated that (1) 2D material can be used as the shell component of a core-shell nanostructure, and (2) surface plasmon can effectively boost sensor performance.

The risk factors for ultrasound-diagnosed non-alcoholic fatty liver disease among adolescents

<p><b>INTRODUCTION</b>Non-alcoholic fatty liver disease (NAFLD) is garnering increasing interest and acceptance as one of the most important causes of chronic liver disease. The aim of this study was to investigate the risk factors for NAFLD among selected adolescent students in Hualien City, Taiwan.</p><p><b>MATERIALS AND METHODS</b>A stratified random sampling scheme was carried out among 1724 adolescent students aged 12 or 13 years old in Hualien City. In total, 220 students (normal: overweight: obese = 97:48:75) agreed to join the study. They underwent physical examination, laboratory tests and ultrasonography examination of the liver. Diagnosis of NAFLD in this study was based on sonographic evidence of a fatty liver and testing negative for serum HBsAg and anti- HCV antibody.</p><p><b>RESULTS</b>Of the 220 participants, 4 were excluded because they tested positive for HBsAg or anti-HCV antibody. NAFLD was detected in 86 (39.8%) out of the 216 subjects. The rate of NAFLD in the adolescents increased progressively from 16.0% in the normal group to 50.5% in the overweight group, and 63.5% among the obese subjects. Compared to their normal counterparts, adolescents with NAFLD had a significantly higher weight, body mass index (BMI), waist circumference, levels of alanine aminotransferase (ALT), triglyceride and nonhigh- density-lipoprotein (non-HDL) cholesterol. However, among the participants with NAFLD, only 20 (23.3%) showed ALT abnormality but there was an increasing trend of ALT abnormality as the severity of fatty liver increased. In addition, the higher ALT, Homeostasis model assessment- insulin resistance (HOMA-IR), cholesterol, triglyceride, and non-HDL levels and lower HDL-C as the severity of fatty liver increased. In a stepwise logistic regression analysis, the most significant factor associated with the presence of NAFLD was weight category. When compared with their normal counterparts, overweight and obese adolescents had a 4.14 and 5.98 times the risk of having NAFLD, respectively. Elevated ALT was the second most important factor as adolescents with elevated ALT were more likely to have NAFLD (odds ratio = 3.32, 95% CI: 1.16 to 9.50). Non-HDL cholesterol level was the third most important factor associated with NAFLD with a 3.81-fold increase in risk incurred for every l n (1 mg/dL) increment.</p><p><b>CONCLUSIONS</b>Obesity, ALT abnormality and elevated non-HDL-cholesterol are risk factors for NAFLD in adolescents. However, only 23.3% of the adolescents with NAFLD showed an abnormality for ALT. Therefore, ALT alone is not a sufficient indicator; and it is recommended that ultrasonography of the liver should be part of the routine health examination of obese adolescents.</p>