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Donation after circulatory death (DCD) can expand the donor pool effectively. A gap remains in outcome between DCD livers and living donor livers, warranting improved DCD liver quality and urgent resolution. Bone marrow mesenchymal stem cells (BMMSCs) can regulate immunity, participate in the anti-inflammatory response, and secrete cytokines. We investigated the effect of BMMSCs combined with normothermic machine perfusion (NMP) on DCD liver quality, and the role of microcirculation therein. Rat thoracic aortas were clipped to obtain DCD livers, and a rat NMP system was established. The DCD livers were grouped by preservation method: normal, static cold storage (SCS), NMP (P), and BMMSCs plus NMP (BP); storage time was up to 8 h. Liver function in outflow perfusate was detected by biochemical methods; liver tissue histopathology was observed by hematoxylin–eosin staining; hepatocyte ultrastructure was observed by transmission electron microscopy; hepatocyte apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling; liver microcirculation–related indicators were detected by immunofluorescence, immunohistochemistry, Western blotting, and enzyme-linked immunosorbent assay. Compared with SCS, P and BP significantly improved liver function and liver histological damage, reduced hepatocyte apoptosis, and repaired hepatocyte mitochondrial damage after 6 h in vitro. BP also significantly inhibited intrahepatic macrophage activation and intercellular adhesion, improved endothelial damage, and significantly improved endothelin 1–nitric oxide balance and microcirculation perfusion. In conclusion, BP can improve DCD liver microcirculation and quality. The mechanism may be the improvement of improve hepatic sinusoidal endothelial injury and microcirculation perfusion by inhibiting macrophage activation and intercellular adhesion. Electronic supplementary material The online version of this article (10.1007/s00441-020-03202-z) contains supplementary material, which is available to authorized users.

Hepatocellular carcinoma (HCC) is the third most common cause of cancer mortality worldwide, which is partially due to the lack of appropriate therapeutic options. The development of HCC is accompanied with unique and continuous genomic and epigenetic modifications. Therefore, the absence of a personalized and reproducible human model reduces the ability to determine the potential of candidate treatments. Conditional reprogramming (CR) culture has been used to establish and indefinitely grow patient‑derived tumor cell lines in a rapid and efficient manner. In the present study, primary HCC cells were isolated from tumor specimens and cultured under CR conditions. The proliferative potential and capacity of cells to undergo continuous regeneration were evaluated by cell viability and proliferation assays, and the expression of tumor‑specific markers was determined by western blotting and immunofluorescence to determine the prospects for use in clinical settings. It was demonstrated that ~55% of tumor samples were able to generate HCC cells that could be continuously expanded and passaged under CR conditions; this ability was associated with the source and composition of the tumor tissues. Furthermore, the expression of the tumor‑specific marker α‑fetoprotein and the proliferative ability of cells were maintained following cycles of cryopreservation and resuscitation. In conclusion, with further optimization, the CR system may be a useful tool for the precise therapeutic treatment of patients with HCC.

The application of Water-Sediment Regulation Project provides abundant freshwater for the Yellow River Delta, changes water and sediment condition, as well as brings lots of exogenous substances. Using orthogonal test with three factors and four levels, we examined the effects of water condition, sediment burial depth and exogenous nitrogen input on the growth of wetland plant, Suaeda salsa. The results showed that sediment burial had great effect on protein content and SOD activity. Nitrogen input had great effect on POD activity. CAT activity was not affected by sediment burial, nitrogen input and water depth. The water depth manipulation had significant effect on leaf, stem and total dry weight. With the increases of water depth, leaf, stem and total dry weight showed a decreasing trend, with the maximum values (25.70, 40.86, 69.73 g) at the 2 cm water depth. There was no effect of nitrogen input and sediment burial on dry weight. The results of range analysis showed that the effect of water depth on leaf, stem, root and total dry weight was great, and followed by nitrogen input and sediment burial, with an optimal combination of 2 cm water depth +12 cm sediment burial + 9 g·m调水调沙工程的实施为黄河三角洲带来大量淡水恢复退化湿地的同时,也改变了湿地的水沙状况并带来大量的外源物质。本研究采用3因素4水平的正交试验,以黄河口滨岸潮滩湿地典型的先锋物种碱蓬为研究对象,探讨了其对淹水深度、泥沙埋深及外源氮输入的适应机制。结果表明: 泥沙埋深对碱蓬叶片蛋白含量及超氧化物歧化酶(SOD)活性的影响较大;氮输入对过氧化物酶(POD)活性的影响较大,而3种处理对过氧化氢酶(CAT)活性的影响无显著差异。淹水深度对叶、茎及总干重具有显著影响,且随淹水深度的增加呈现减小趋势,最大值(25.70、40.86、69.73 g)均出现在2 cm淹水处理;而氮输入及泥沙埋深对碱蓬干重的影响不显著。极差分析表明,淹水深度对碱蓬叶、茎、根及总干重的影响最为显著,其次为氮输入和泥沙埋深,2 cm淹水深度+12 cm埋深深度+9 g·m

Background Donation after circulatory death (DCD) liver grafts have a poor prognosis after transplantation. We investigated whether the outcome of DCD donor organs can be improved by heme oxygenase 1 (HO-1)-modified bone marrow-derived mesenchymal stem cells (BMMSCs) combined with normothermic machine perfusion (NMP), and explored its underlying mechanisms. Methods BMMSCs were isolated, cultured, and transduced with the HO-1 gene. An NMP system was established. DCD rat livers were obtained, preserved by different methods, and the recipients were divided into 5 groups: sham operation, static cold storage (SCS), NMP, BMMSCs combined with NMP, and HO-1/BMMSCs combined with NMP (HBP) groups. Rats were sacrificed at 1, 7, and 14 days after surgery; their blood and liver tissue samples were collected; and liver enzyme and cytokine levels, liver histology, high-mobility group box 1 (HMGB1) levels in monocytes and liver tissues, and expression of Toll-like receptor 4 (TLR4) pathway-related molecules were evaluated. Results After liver transplantation, the SCS group showed significantly increased transaminase levels, liver tissue damage, and shorter survival time. The HBP group showed lower transaminase levels, intact liver morphology, prolonged survival time, and decreased serum and liver proinflammatory cytokine levels. In the NMP and SCS groups, HMGB1 expression in the serum, monocytes, and liver tissues and TLR4 pathway-related molecule expression were significantly decreased. Conclusions HO-1/BMMSCs combined with NMP exerted protective effects on DCD donor liver and significantly improved recipient prognosis. The effect of HO-1/BMMSCs was greater than that of BMMSCs and was mediated via HMGB1 expression and TLR4 pathway inhibition.

There is a need to improve the quality of donor liver from donation after circulatory death (DCD). The purpose of this study was to investigate the effects and mechanism of normothermic machine perfusion (NMP) combined with bone marrow mesenchymal stem cells (BMMSCs) on the oxidative stress and mitochondrial function in DCD livers. DCD livers were obtained, a rat NMP system was established, and BMMSCs were extracted and identified. The DCD livers were grouped by their preservation method: Normal, static cold storage (SCS), NMP (P), and NMP combined with BMMSCs (PB), and the preservation time was up to 8 h. An IAR20 cell oxidative stress injury model was established in vitro by simulating DCD oxidative stress injury and coculturing with BMMSCs for 6 h. Compared with SCS group, after 6 h in vitro, the PB and P groups had significantly improved liver function and liver histological damage, reduced hepatocyte apoptosis and oxidative stress, improved hepatocyte mitochondrial damage, and increased mitochondrial membrane potential. These indicators were significantly better in the PB group than in the P group. BMMSCs significantly inhibited reactive oxygen species release from the IAR20 cell oxidative stress model in vitro, ameliorated mitochondrial damage, and increased mitochondrial membrane potential level. BMMSCs also downregulated the JUN N-terminal kinase-nuclear factor kappa B (JNK-NF-κB) signaling pathway significantly in the IAR20 cell oxidative stress model and promoted AMP-activated protein kinase (AMPK) activation. We verified that NMP combined with BMMSCs also played the same role in the PB group. NMP combined with BMMSCs could improve liver quality by relieving oxidative stress injury and improving mitochondrial function in rat DCD livers. The mechanism of protective role might involve inhibiting the JNK-NF-κB pathway to reduce oxidative stress and promote AMPK activation, thereby reducing mitochondrial damage and increase mitochondrial function.

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-β 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.

The aim of the present study was to explore the effects of co-culturing bone marrow-derived mesenchymal stem cells (BM-MSCs) cultured with hepatitis B virus (HBV)-infected lymphocytes in vitro. BM-MSCs 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, BM-MSCs + HepG2.2.15 cells; group 4, SLCs + HepG2.2.15 cells; and group 5, SLCs + BM-MSCs + 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 enzyme-linked immunosorbent assay (ELISA). T cell subsets were quantified by flow cytometry using fluorescence-labeled 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 interferon-γ was negatively correlated with the level of HBV DNA, whereas secretion of interleukin (IL)-10 and IL-22 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 co-cultured with BM-MSCs, SLCs, or in the two types of cells combined. BM-MSCs 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.

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.

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.

The purpose of the present study was to investigate whether bone marrow mesenchymal stem cells (BMMSCs) modified by CXC‑chemokine receptor type 3 (CXCR3) and heme oxygenase‑1 (HO‑1) genes can repair damaged intestinal epithelial cells in vitro, and the role of the p38 mitogen‑activated protein kinase (p38‑MAPK) pathway in this process. A model of intestinal epithelial crypt cell line‑6 (IEC‑6) damage was created, and BMMSCs were transfected with either the CXCR3 and/or HO‑1 gene in vitro. There were nine experimental groups in which the damaged IEC‑6 cells were co‑cultured with differentially‑treated BMMSCs and lymphocytes for 24 h. Reverse transcription‑quantitative polymerase chain reaction analysis, immunohistochemistry and a western blot analysis were performed to detect stem cell transfection, the repair of damaged intestinal epithelial cells and the expression of related molecules in the P38‑MAPK pathway, respectively. Crystal violet staining and live cell imaging were used to detect the chemotaxis of BMMSCs. Flow cytometry was used to detect T lymphocyte activity and the surface markers expressed on BMMSCs. An ELISA was used to quantify cytokine production. The adenovirus (Ad)‑CXCR3/MSCs exhibited the characteristics of stem cells and exhibited chemotaxis. The Ad‑CXCR3/MSCs and Ad‑(CXCR3 + HO)/MSCs exhibited increased expression of tight junction protein zonula occludens‑1 (ZO‑1) and anti‑proliferating cell nuclear antigen in the damaged IEC‑6 cells, and apoptosis of the damaged IEC‑6 cells was decreased. BMMSCs inhibited the phosphorylation of p38, in addition to downstream molecules of the p38MAPK signaling pathway. The Ad‑CXCR3/MSCs and Ad‑(CXCR3 + HO)/MSCs exhibited significantly decreased expression levels of downstream molecules, including phosphorylated (p)‑p38, p‑activated transcription factor 2, p‑C/EBP homologous protein‑10, and p‑myocyte enhancer factor 2C, and target molecules (e.g., apoptotic bodies). The effects of Ad‑(CXCR3 + HO)/MSCs on the repair of the damaged intestinal tract and inhibition of the p38‑MAPK pathway was more marked than those in other groups on day 7 post‑surgery in the rejection model for small bowel transplantation. BMMSCs modified by the CXCR3 and HO‑1 genes exhibited superior ability to repair damaged intestinal epithelial cells and served this role via the p38‑MAPK pathway.

// Tao Liu 1, 2 , Cai-Hua Zu 3 , Shu-Sen Wang 1, 2 , Hong-Li Song 2 , Zheng-Lu Wang 4, 5 , Xin-Nv Xu 1 , Hong-Sheng Liu 1 , Yu-Liang Wang 1, 6 , Zhong-Yang Shen 1, 2 1 Key Laboratory for Critical Care Medicine of the Ministry of Health, Tianjin First Central Hospital, Tianjin, China 2 Organ Transplant Center, Tianjin First Central Hospital, Tianjin, China 3 First Center Clinical College, Tianjin Medical University, Tianjin, China 4 Department of Pathology, Tianjin First Central Hospital, Tianjin, China 5 Biobank of Tianjin First Central Hospital, Tianjin, China 6 Clinical Laboratory, Tianjin First Central Hospital, Tianjin, China Correspondence to: Yu-Liang Wang, email: wang_yu_l@163.com Zhong-Yang Shen, email: shenzy_2014@163.com Keywords: microRNA, competing endogenous RNA, hepatocellular carcinoma, CD151, PIK3C2A Received: October 12, 2015 Accepted: May 09, 2016 Published: May 30, 2016 ABSTRACT Competing endogenous RNAs (ceRNAs) are RNA transcripts that can crosstalk with each other by competing for shared microRNAs (miRNAs) through miRNA response elements (MREs). Involved in ceRNA networks, the RNA transcripts may be in a balance, disruption of which could lead to tumorigenesis. Here we reveal a ceRNA interaction between PIK3C2A and CD151 mRNAs in hepatocellular carcinoma (HCC) cells. PIK3C2A is a candidate ceRNA of CD151 because mRNA 3′ untranslated regions (3′UTRs) of these two genes contain miR-124 binding sites. miR-124 is downregulated, while PIK3C2A and CD151 are upregulated in HCC cells compared with normal hepatocytes. Direct and negative regulation of PIK3C2A and CD151 by miR-124 was confirmed in HCC cells. miR-124 and the two potential ceRNAs are all recruited to the RNA-induced silencing complex (RISC). In HCC cell lines QGY- 7703 and SMMC-7721, and normal hepatic cell line HL-7702, miR-124 plays a tumor suppressor role by targeting PIK3C2A and CD151. The MREs within PIK3C2A 3′UTR can independently stimulate CD151 expression level by acting as miR-124 decoys. PIK3C2A MREs enhance HCC cell malignancy by absorbing endogenous miR-124 and activating CD151 in HCC cells. We conclude that PIK3C2A 3’UTR functions as a trans activator to stimulate CD151 by competing for miR-124 binding in HCC cells. The collaboration of PIK3C2A and CD151 through ceRNA mechanism may be implicated in HCC initiation and development.

The Yellow River Delta has been facing the threat of functional degradation during the recent years. The Water-Sediment Regulation Project not only supplements abundant freshwater, but also alters the sediment burial and heavy metal levels, which affects vegetation growth. Thus, we selected the pioneer species

Plunge milling cutter withstanding greater loads in heavy milling can cause tool breakage, cutting vibration, even chatter. Firstly, geometric model of plunge milling cutter on various combinations of axial rake angle and radial rake angle are established. Secondly, with combination different axial rake angle and radial rake angle of plunge milling cutter under static load, the amount of deformation plunge milling cutter are calculated by finite element analysis. Finally, the plunge milling cutter first six natural frequencies are given in plunge milling cutter modal analysis with finite element analysis. The results show that minimum total deformation of plunge milling cutter is given with the axial rake angle 5° and radial rake angle-5 ° of the cutter and when the tool working at frequencies up to 10012HZ has caused vibration within the XY plane.

The objective of this study was to explore the effects of dendritic cells (DCs) from hepatitis B virus (HBV) transgenic mice-stimulated autologous lymphocytes on in vitro HBV replication. DCs from HBV transgenic mice were induced to maturity by lipopolysaccharide, followed by incubation with hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) in vitro. Mature DCs and autologous lymphocytes were co-stimulated to form specific sensitized immune effector cells (IEC), which were then co-cultured with the human hepatoma cell line HepG2.2.15. Changes in morphology and activity of hepatocytes were then observed, as well as analysis of changes in liver enzyme, and HBV DNA and inflammatory cytokine levels in the culture supernatant. Intracellular HBV DNA and covalently closed circular DNA (cccDNA) concentration were measured by real-time polymerase chain reaction. Co-stimulation by mature DCs and IEC showed no impact on the morphology and liver enzyme expression level of HepG2.2.15 cells, but the supernatant HBV DNA and intracellular HBV DNA and cccDNA levels decreased significantly compared with those cells co-cultured with immature DCs. Secretion of inflammatory cytokines in the supernatant showed that when HBV DNA was highly expressed, the concentration of IFN-γ and IL-2 decreased, while IL-10 increased. Contrastingly, when HBV DNA had low expression, the concentration of IFN-γ and IL-2 increased and IL-10 decreased. Co-stimulation of HBV-related antigen-induced mature DCs and autologous lymphocytes showed inhibitory effects on ex vivo HBV replication, and cytokines were suggested to mediate this effect.

In recent years, the smartphones which are integrated with many high-accuracy sensors have become very popular. Utilizing the development of sensors, developers have done many useful researchers. Indoor positioning navigation is an interesting and hot field of them. Step detecting and counting are key technologies in indoor positioning. In this paper, we propose a novel method to count steps which is an improvement of the classical steps detection method, the peak detection method of the acceleration. Our proposed scheme consists of two parts: the scoring part and adaptive window length part. Two adaptive window length algorithms which can adapt to the varying velocity are also proposed in this paper. In our experiments, a smartphone - Nubia NX513J is handled, with screen facing upwards to record the accelerometer data. The scoring algorithm of acceleration in the process of step detection is tested in two kinds of path: a straight path and a U-shaped path in which it shows better results than conventional peak detection method.

As a group of heterogeneous multipotent cells, mesenchymal stem cells (MSCs) have potential in treatment of a variety of clinical diseases. However, the low survival of the transplanted MSCs reduced their therapeutic effects. In this study, we revealed that rno-miR-203 suppressed activity and colony formation and enhanced apoptosis of the rat bone marrow-derived MSCs (BM-MSCs). Using bioinformatics analysis, we found a potential miR-203 binding site within rat phosphatidylinositol 3-kinase (PI3K) 3'UTR, and fluorescent reporter experiments validated the direct and negative regulation of PI3K expression by miR-203 through this site. Ectopic expression of PI3K rescued BM-MSCs from depressed activity induced by miR-203, and suppression of PI3K attenuated the increased BM-MSCs activity by miR-203 inhibitor treatment. Moreover, miR-203 blocking partly protected BM-MSCs from impairment caused by low nutrition. We conclude that inhibition of endogenous miR-203 elevated PI3K expression, which may strengthen PI3K/Akt pathway and promote BM-MSCs activity and survival. © 2014 IUBMB Life, 66(3):220-227, 2014.

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.

Background Cerebral oedema leading to cerebral herniation is a major cause of death during acute liver failure (ALF), but the underlying mechanism is not clear. Aims We investigated the role of tumour necrosis factor (TNF)-alpha in changing the permeability of the blood-brain barrier (BBB) during ALF. Methods ALF animal models were generated by administering D-galactosamine (GalN) and lipopolysaccharide, or GalN and TNF-alpha. ALF induction was blocked by first administering anti-TNF-alpha-IgG or anti-TNF-alpha-R1. We investigated the BBB permeability with Evans blue staining, and the structure with electron microscopy. Results BBB permeability increased in ALF mice and correlated with elevated serum TNF-alpha levels. No vascular endothelial cell (EC) apoptosis was detected, but electron microscopy of cells from human and mouse ALF tissues revealed tight junction (TJ) disruptions and EC shrinkage, as well as increased vesicles and vacuoles. In addition, the expression of the TJ-associated protein occludin was significantly decreased in both ALF mice and patients, although the expression of occludin mRNA did not change. Changes in BBB permeability, brain tissue ultrastructure and occludin expression in ALF-induced mice could be prevented by prophylaxis treatment with either antibody to TNF-alpha-IgG or antibody to TNF-alpha-R1. Conclusions Our results suggest that TNF-alpha plays a critical role in the development of brain oedema in ALF, and that both vasogenic and cytotoxic mechanisms may be involved. Increased BBB permeability may be because of the disruption of TJs, and loss of the TJ-associated protein occludin.

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-βlevels were significantly increased, while IL-2, IL-6, IL-17, IL-23, tumor necrosis factor-α, and interferon-γlevels 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.

Anti-HBV therapy is essential for patients awaiting liver transplantation. This study aimed to explore the effects of dendritic cells (DCs) derived from the peripheral blood of hepatitis B patients on the replication of HBV in vivo and to evaluate the biosafety of DCs in clinical therapy. Peripheral blood mononuclear cells (PBMCs) were isolated from HBV-infected patients and maturation-promoting factors and both HBsAg and HBcAg were used to induce DC maturation. Mature DCs and lymphocytes were co-cultured with human hepatocyte cell HL-7702 or HBV-producing human hepatocellular carcinoma cell HepG2.2.15. We found that mature lymphocytes exposed to DCs in vitro did not influence morphology or activities of HL-7702 and HepG2.2.15 cells. Liver function indexes and endotoxin levels in the cell supernatants did not change in these co-cultures. Additionally, supernatant and intracellular HBV DNA levels were reduced when HepG2.2.15 cells were co-cultured with mature lymphocytes that had been cultured with DCs, and HBV covalently closed circular DNA (cccDNA) levels in HepG2.2.15 cells also decreased. Importantly, DC-mediated immunotherapy had no mutagenic effect on HBV genomic DNA by gene sequencing of the P, S, X, and C regions of HBV genomic DNA. We conclude that PBMC-derived DCs from HBV-infected patients act on autologous lymphocytes to suppress HBV replication and these DC clusters showed favorable biosafety.

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

To investigate the effects of nucleoside analogues on hepatitis B virus (HBV) in hepatic lymph nodes of hepatitis B related liver transplantation recipients who were hepatitis B surface antigen (HBsAg) positive but negative for serum HBV DNA.From June 2010 to March 2011, thirty-six cases of hepatitis B related liver transplantation recipients [32 males, 4 females, average age (54 ± 7) years] were divided into drug treatment group and non-drug treatment group according to the utility of nucleoside analogues. Drug treatment group was divided into two subgroups: drug treatment3 months group and drug treatment ≤ 3 months group. The hepatic lymph nodes in the hepatoduodenal ligament were taken during the operation of liver transplant. Using nested or semi-nested PCR, HBV DNA and the replicative form HBV cccDNA in hepatic lymph nodes were detected. Data were analyzed by Fisher's exact test.The positive rate of HBV DNA: the difference was not statistically significant between drug treatment group (72.7%, 16/22) and non-drug treatment group (14/14) (P = 0.062), the difference was not statistically significant between drug treatment3 months group (10/14) and drug treatment ≤ 3 months group (6/8) in the subgroups of drug treatment group (P = 1.000). The positive rate of HBV cccDNA: drug treatment group (22.7%, 5/22) was significantly lower than the non-drug treatment (12/14) (P = 0.000), drug treatment3 months group (1/14) was significantly lower than drug treatment ≤ 3 months group (4/8) in the subgroups of drug treatment group (P = 0.039).Hepatic lymph nodes maybe one of the extrahepatic HBV reservoirs. Treating with nucleoside analogues more than 3 months can significantly decrease the replication of HBV in hepatic lymph nodes of HBV associated liver transplantation recipients.

To provide a basis for improved prevention and treatment of hepatitis B virus (HBV) re-infection after liver transplantation, variations in the S and P genes of HBV under immunosuppression in vitro and their association with patient prognosis were investigated. For the in vitro study, HepG2.2.15 hepatocellular carcinoma cells stably producing HBV particles were treated with the immunosuppressants methylprednisolone (MP) and tacrolimus (FK506) at doses found to be non-toxic by the methylthiazolyl tetrazolium (MTT) cell viability assay. MP dose-dependently inhibited HBV DNA expression in HepG2.2.15 cells, while FK506 did not, as determined by quantitative real-time PCR (qRT-PCR). By gene sequencing, both MP and FK506 were found to cause variations in HBV S, P, and S/P overlapping regions. MP- but not FK506-induced mutations were common in the glucocorticoid response element of the P region, while both immunosuppressants caused mutations outside the nucleoside analogue resistance sites. For the in vivo study, 14 patients with HBV-related end-stage liver disease re-infected after liver transplantation, and 20 cases without HBV re-infection as controls, were studied. Seventy-five percent of re-infected recipients showed multi-loci amino acid mutations at different sites besides lamivudine (LAM)-resistant loci in the P region, including in the glucocorticoid response element. Fifty percent of re-infected recipients had mutations in the "a" determinant region and flanking sequences. Re-infection was associated with negative serum hepatitis B immunoglobulin (HBIG), as measured by a microparticle capture enzyme immunoassay. Nucleotide mutations in the S region caused missense or synonymous mutations, which caused synonymous mutations in the overlapping P region. These results showed that effects of immunosuppressants on HBV genes in vitro were different from those in clinical recipients. Positive HBV DNA and gene mutations pre-transplantation were factors affecting re-infection post-transplantation. Multiple mutations found in the P and S genes suggest that the formation of quasispecies contributes to HBV re-infection after liver transplantation.

The characteristics of methane (CH4) fluxes from tidal wetlands of the Yellow River estuary were observed in situ with static-chamber and GC methods in September and October 2009, and the key factors affecting CH4 fluxes were discussed. From the aspect of space, the CH4 flux ranges in high tidal wetland, middle tidal wetland, low tidal wetland, bare flat are - 0.206-1.264, -0.197-0.431, -0.125-0.659 and -0.742-1.767 mg x (m2 x h)(-1), the day average fluxes are 0.089, 0.038, 0.197 and 0.169 mg x (m2 x h)(-1), respectively, indicating that the tidal wetlands are the sources of CH4 and the source function of CH4 differed among the four study sites, in the order of low tidal wetlandbare flathigh tidal wetlandmiddle tidal wetland. From the aspect of time, the ranges of CH4 fluxes from the tidal wetland ecosystems are -0.444-1.767 and - 0.742- 1.264 mg x (m2 x h)(-1), and the day average fluxes are 0.218 and 0.028 mg x (m2 x h)(-1) in September and October, respectively. The CH4 fluxes in each tidal wetland in September are higher than those in October except that the high tidal wetland acts as weak sink in September. Further studies indicate that the changes of environmental factors in the Yellow River estuary are complicated, and the CH4 fluxes are affected by multiple factors. The differences of CH4 fluxes characteristics among different tidal wetlands in autumn are probably related to temperature (especially atmospheric temperature) and vegetation growth status, while the effects of water or salinity condition and tide status on the CH4 flux characteristics might not be ignored.

The increasing demand for transplantation has led application of steatotic liver as the graft. The aim of this study was to determine the effect of donor graft steatosis on overall outcome and tumor recurrence after liver transplantation for hepatocellular carcinoma.131 patients that underwent liver transplantation for hepatocellular carcinoma between 2007 and 2008 were included. Donor steatosis was categorized as non-steatosis group (0%-10%, n=101) and steatosis group (10%, n=30). The Kaplan-Meier method and Cox proportional hazard regression model was used for data analysis.Postoperative recipient survival rate was 81% and 66.6% at 1 and 3 years, respectively, for non-steatotic graft; 87.5% and 58.3% for mild steatosis; 83.3% and 41.7% for moderate to severe steatosis (p=0.303). Postoperative tumor recurrence rate was 15.8% and 28.7% at 1 and 3 years, respectively, for grafts with no steatosis; 8.3% and 20.8% for those with mild steatosis; 33.3% and 50% for those with moderate to severe steatosis, (p0.05).Steatotic donor was not associated with a worse prognosis in early stage postoperative and mild fatty liver did not increase tumor recurrence risks. The moderate to severe status of fatty liver had some effect on tumor recurrence.

Cerebral oedema leading to cerebral herniation is a major cause of death during acute liver failure (ALF), but the underlying mechanism is not clear.We investigated the role of tumour necrosis factor (TNF)-alpha in changing the permeability of the blood-brain barrier (BBB) during ALF.ALF animal models were generated by administering D-galactosamine (GalN) and lipopolysaccharide, or GalN and TNF-alpha. ALF induction was blocked by first administering anti-TNF-alpha-IgG or anti-TNF-alpha-R1. We investigated the BBB permeability with Evans blue staining, and the structure with electron microscopy.BBB permeability increased in ALF mice and correlated with elevated serum TNF-alpha levels. No vascular endothelial cell (EC) apoptosis was detected, but electron microscopy of cells from human and mouse ALF tissues revealed tight junction (TJ) disruptions and EC shrinkage, as well as increased vesicles and vacuoles. In addition, the expression of the TJ-associated protein occludin was significantly decreased in both ALF mice and patients, although the expression of occludin mRNA did not change. Changes in BBB permeability, brain tissue ultrastructure and occludin expression in ALF-induced mice could be prevented by prophylaxis treatment with either antibody to TNF-alpha-IgG or antibody to TNF-alpha-R1.Our results suggest that TNF-alpha plays a critical role in the development of brain oedema in ALF, and that both vasogenic and cytotoxic mechanisms may be involved. Increased BBB permeability may be because of the disruption of TJs, and loss of the TJ-associated protein occludin.

Background Spontaneous bacterial peritonitis (SBP) is a common clinical disease and one of the most severe complications of acute liver failure (ALF). Although the mechanism responsible for SBP is unclear, cytokines play an important role. The aim of this study was to investigate the effects of tumor necrosis factor-alpha (TNF-α) on the structure of the intestinal mucosa and the expression of tight junction (Zona Occludens 1; ZO-1) protein in a mouse model of ALF. Methods We induced ALF using D-galactosamine/lipopolysaccharide (GalN/LPS) or GalN/TNF-α and assessed the results using transmission electron microscopy, immunohistochemistry, Western blotting, ELISA and real-time quantitative PCR. The effects of administration of anti-TNF-α IgG antibody or anti-TNF-α R1 antibody before administration of GalN/LPS or GalN/TNF-α, respectively, on TNF-α were also assessed. Results Morphological abnormalities in the intestinal mucosa of ALF mice were positively correlated with serum TNF-α level. Electron microscopic analysis revealed tight junction (TJ) disruptions, epithelial cell swelling, and atrophy of intestinal villi. Gut bacteria invaded the body at sites where TJ disruptions occurred. Expression of ZO-1 mRNA was significantly decreased in both ALF models, as was the level of ZO-1 protein. Prophylactic treatment with either anti-TNF-α IgG antibody or anti-tumor necrosis factor-a receptor1 (anti-TNF-α R1) antibody prevented changes in intestinal tissue ultrastructure and ZO-1 expression. Conclusion TNF-α affects the structure of the intestinal mucosa, decreases expression of ZO-1, and affects the morphology of the colon in a mouse model of ALF. It also may participate in the pathophysiological mechanism of SBP complicated to ALF.

To explore the alterations of intestinal mucosa morphology, and the effects of tumor necrosis factor alpha (TNFalpha) on enterocyte apoptosis in mice with fulminant hepatic failure (FHF).Liver damage was induced by lipopolysaccharide (LPS)/TNF-alpha in D-galactosamine (GalN) sensitized BALB/c mice. There were 40 mice in normal saline (NS)-treated group, 40 mice in LPS-treated group, 40 mice in GalN-treated group, 120 mice in GalN/ LPS-treated group and 120 mice in GalN/TNFalpha-treated group. Each group was divided into five subgroups of eight mice each. Serum samples and liver, intestinal tissues were respectively obtained at 2, 6, 9, 12 and 24 h after administration. Anti-TNFalpha monoclonal antibody was injected intravenously into GalN/LPS-treated mice. Serum TNFalpha levels were determined by enzyme-linked immunosorbent assays (ELISA). Serum ALT levels were determined using an automatic analyzer. The intestinal tissues were studied under light microscope and electron microscope at 2, 6, 9, 12 and 24 h in mice with fulminant hepatic failure, respectively. Enterocyte apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) method. The expression of tumor necrosis factor receptor 1 (TNFR1) in intestinal tissue was tested by immunohistochemistry Envision Two Steps.Gut mucosa was morphologically normal at all time points in all groups, but typical apoptotic cells could be seen in all experimental groups under electron micro-scope. Apoptosis rate of gut mucosal epithelial cells were significantly increased at 6, 9 and 12 h, peaked at 12 h in mice with fulminant hepatic failure. TNFalpha induced apoptosis of enterocytes in mice with FHF. The integrated OD (IOD) levels of TNFalpha receptor 1 protein expressed in the intestine of mice with GalN/LPS and GalN/ TNFalpha-induced FHF at 2, 6, 9, 12 and 24 h after GalN/LPS and GalN/TNFalpha administration were 169.54+/-52.62/905.79+/-111.84, 11,350.67+/-2 ,133.26/28,160.37+/-4,601.67, 25,781.00 +/-2,277.75/122,352.30+/-49,412.40, 5,241.53+/-3,007.24/49,157.93+/-9,804.88, 7,086.13+/-1,031.15/3,283.45+/-127.67, respectively, compared with those in control groups (with NS, LPS and GalN administration, respectively). IOD level of TNFR1 changed significantly at 6, 9 and 12 h after GalN/LPS and GalN/TNFalpha administration. The expression of TNFR1 protein was significantly higher at 9 h after GalN/LPS and GalN/TNFalpha administration than that in control groups. Protein expression of TNFR1 was positively correlated with enterocyte apoptosis.TNFalpha can induce apoptosis of enterocytes in mice with FHF. Anti-TNFalpha IgG can inhibit this role.

To study the role of tumor necrosis factor-alpha (TNFalpha) on enterocyte apoptosis in the experimental model of fulminant hepatic failure (FHF).Liver damage was induced by lipopolysaccharide (LPS)/TNFalpha in D-galactosamine (GalN) sensitized BALB/c mice. Serum TNFalpha levels were determined by enzyme-linked immunosorbent assays (ELISA). The intestinal tissues were studied micro- and ultra-microscopically at 2 h, 6 h, 9 h, 12 h and 24 h time points in mice with fulminant hepatic failure. Enterocyte apoptosis was determined by TUNEL method. The TNFR I expression in the intestinal tissue was tested by immunohistochemistry.(1) Gut mucosa was morphologically normal at every time point in all groups, but typical apoptotic cells could be seen in the experimental groups under the electron microscope. Apoptosis rate of gut mucosal epithelial cells was significantly increased at 6 h (large intestine: 6.47e(-3)+/-2.91e(-4); small intestine: 6.64e(-3)+/-3.78e(-4)), 9 h (large intestine: 6.81e(+4)+/-7.41e(+3); small intestine: 2.58e(+4)+/-2.28e(+3)) and 12 h (large intestine: 4.92e(+4)+/-9.80e(+3); small intestine: 5.24e(+4)+/-3.01e(+3)), and peaked at 12 h in mice with FHF. (2) TNFalpha induced apoptosis of enterocytes in mice with FHF. Anti-TNFalpha inhibited this effect. (3) The integrated OD (IOD) levels of TNFalpha receptor I protein expressed differently in the intestine of mice with GalN/LPS and GalN/ TNFalpha-induced FHF at 9 h after GalN/LPS and GalN/ TNFalpha administration, in comparison with those of the control groups. IOD level of TNFRI changed significantly at 6 h (large intestine: 2.82e(+4)+/-4.60e(+3); small intestine: 1.14e(+4)+/-2.13e(+3)), 9 h (large intestine: 6.81e(+4)+/-7.41e(+3); small intestine: 2.58e(+4)+/-2.28e(+3)) and 12 (large intestine: 4.92e(+4)+/-9.80e(+3); small intestine: 5.24e(+4)+/-3.01e(+3)) hours after GalN/LPS and GalN/ TNFa administration. The expression of TNFR1 protein was significantly higher at 9 and 12 h after GalN/LPS and GalN/TNFa administration than other time points. Protein expression of TNFR1 was positively correlated with enterocyte apoptosis.TNFa can induce enterocyte apoptosis in mice with FHF. Anti- TNFalpha IgG can inhibit this role. Excessive TNFRI expression of enterocyte in fulminant hepatic failure can be induced by TNFa, which suggests that TNFalpha can induce apoptosis of enterocyte by up-regulation of TNFRI protein expression.

AIM: To explore the protective effect of bone marrow mesenchymal stem cells (BM MSCs) in the small intestinal mucosal barrier following heterotopic intestinal transplantation (HIT) in a rat model. METHODS: BM MSCs were isolated from male Lewis rats by density gradient centrifugation, cultured, and analyzed by flow cytometry. The HIT models were divided into a non-rejection group, saline-treated rejection group (via penile vein), and BM MSC–treated group (via penile vein). Intestinal mucosal barrier injury was estimated by diamine oxidase (DAO) and D-lactic acid (D-LA) expression levels. Tumor necrosis factor-α (TNF-α), interferon-γ (INF-γ), interleukin-10 (IL-10), and transforming growth factor-β (TGF-β) were detected by enzyme-linked immunosorbent assay. Ultrastructural change of tight junctions (TJs) was observed under transmission electron microscope. Expression levels of the TJ proteins occludin and zona occludens (ZO)-1, affected by the inflammatory factors, were measured using real-time polymerase chain reaction and Western blotting. RESULTS: The pathological score at each time point after surgery indicated significantly less serious injury in the BM MSCs-treated group than in the rejection group (P < 0.05). In the former, graft levels of DAO and D-LA were reduced, and TNF-α and INF-γ production was inhibited (at day 7: 10.6473 ± 0.0710 vs 17.2128 ± 0.4991, P < 0.05; 545.1506 ± 31.9416 vs 810.2637 ± 25.1175, P < 0.05). IL-10 and TGF-β production was increased greatly (at day 7: 125.7773 ± 4.7719 vs 80.3756 ± 2.5866, P < 0.05; 234.5273 ± 9.3980 vs 545.1506 ± 31.9416, P < 0.05). There was increased expression of occludin and ZO-1 protein (at day 7: 0.2674 ± 0.0128 vs 0.1352 ± 0.0142, P < 0.05; at day 5: 0.7189 ± 0.0289 vs 0.4556 ± 0.0242, P < 0.05) and mRNA (at day 7: 0.3860 ± 0.0254 vs 0.1673 ± 0.0369, P < 0.05; at day 5: 0.5727 ± 0.0419 vs 0.3598 ± 0.0242, P < 0.05). CONCLUSION: BM MSCs can improve intestinal barrier permeability, repair TJs, and increase occludin and ZO-1 protein expression. With altered cytokine levels, they can protect the intestinal mucosa after transplantation.

AIM: To investigate the effect of bone-marrow mesenchymal stem cells (BM MSCs) on the intestinal mucosa barrier in ischemia/reperfusion (I/R) injury. METHODS: BM MSCs were isolated from male Sprague-Dawley rats by density gradient centrifugation, cultured, and analyzed by flow cytometry. I/R injury was induced by occlusion of the superior mesenteric artery for 30 min. Rats were treated with saline, BM MSCs (via intramucosal injection) or tumor necrosis factor (TNF)-α blocking antibodies (via the tail vein). I/R injury was assessed using transmission electron microscopy, hematoxylin and eosin (HE) staining, immunohistochemistry, western blotting and enzyme linked immunosorbent assay. RESULTS: Intestinal permeability increased, tight junctions (TJs) were disrupted, and zona occludens 1 (ZO-1) was downregulated after I/R injury. BM MSCs reduced intestinal mucosal barrier destruction, ZO-1 downregulation, and TJ disruption. The morphological abnormalities after intestinal I/R injury positively correlated with serum TNF-α levels. Administration of anti-TNF-α IgG or anti-TNF-α receptor 1 antibodies attenuated the intestinal ultrastructural changes, ZO-1 downregulation, and TJ disruption. CONCLUSION: Altered serum TNF-α levels play an important role in the ability of BM MSCs to protect against intestinal I/R injury.

AIM: To study the permeability of the blood brain barrier (BBB) in a mouse model of acute liver necrosis. METHODS: Male Balb/c mice were divided into 4 groups. In one group, mice were intraperitoneal of lipopolysaccharide (LPS, 10 μg/kg) with D-galactosamine (GalN, 800 mg/kg) to induce acute liver necrosis. Other groups were controls. Serum levels of alanine transaminase (ALT) were determined and the liver tissues were fixed for histopathological analysis. The permeability of BBB in mice was investigated with Evans blue (EB). RESULTS: The serum levels of ALT were increased mildly in mice, which were administration of LPS or GalN alone. And no animals died. But the levels of ALT began to increase at 6 hours (41.89±14.57 μat/L), and reached a maximal level at 12 hours (170.30±16.13 μat/L) after injection with both LPS and GalN. Mice began to die at 6 hours, and at 9 hours after injection, the rate of lethality reached an extremely high level of 66.6%. The liver became massive or submassive necrosis. The concentration of EB in brain was significantly increased in ALF models compared with other groups. CONCLUSION: The permeability of BBB is increased in acute liver necrosis model. It may be the mechanism of the brain edema.