Effects of Rosa roxburghii Tratt on Ulcerative Colitis: An Integrated Analysis of Network Pharmacology and Experimental Validation.

Rosa roxburghii Tratt is a traditional Chinese plant that has been used to treat different inflammatory diseases. The purpose of this study was to investigate the mechanism of action of Rosa roxburghii Tratt extract (RRTE) against ulcerative colitis (UC) using network pharmacology and experimental validation. HPLC-Q/Orbitrap MS was used to rapidly identify the substances contained in RRTE after extracting the active components from the fruit. Then, network pharmacology combined with molecular docking was used to explore the critical target and potential mechanism of RRTE against UC using the active ingredients in RRTE as the research object. Data are presented in a visual manner. Finally, the pharmacological effects of RRTE in alleviating UC were further verified using a DSS-induced UC model of NCM460. The results showed that 25 components in RRTE were identified. A total of 250 targets of the active components and 5376 targets associated with UC were collected. Furthermore, a systematic analysis of the Protein-Protein Interaction (PPI) networks suggests that epidermal growth factor receptor (EGFR), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), and serine/threonine kinase 1 (AKT1) are critical targets for RRTE in the treatment of UC. A comprehensive regulatory network analysis showed that RRTE alleviated UC through the EGFR-mediated PI3K/Akt pathway, and molecular docking showed that active components could strongly bind to EGFR, PIK3R1, and AKT1. In addition, RRTE alleviated dextran sulfate sodium salt (DSS)-induced cell injury and significantly decreased the protein expression levels of EGFR, PIK3R1, and p-AKT in NCM460 cells in vitro. Furthermore, RRTE significantly regulated the expression of the apoptosis-related proteins Apoptotic protease-activating factor 1 (Apaf1), cleaved caspase-3, B-cell lymphoma-2 (Bcl2), and Bcl2 associated X protein (Bax). In conclusion, the components of RRTE are complex, and RRTE can relieve UC through the EGFR-mediated PI3K/Akt pathway.

Transcriptional adaptor 3 influences the proliferative and invasive phenotypes of non-small cell lung cancer cells via regulating EMT.

Transcriptional adaptor 3 (TADA3/ADA3) is a conserved transcriptional co-activator and is dysregulated in many aggressive tumors. However, the role of TADA3 in non-small cell lung cancer (NSCLC) remains unknown. It was previously demonstrated that TADA3 expression correlates with poor prognosis in patients with NSCLC. In the present study, the expression and function of TADA3 were investigated in cells in vitro and in vivo. TADA3 expression was evaluated in clinical specimens and cell lines using reverse transcription-quantitative PCR and western blot analysis. The TADA3 protein level was significantly higher in human NSCLC specimens compared with matched normal tissues. In human NSCLC cell lines, short hairpin RNA-mediated silencing of TADA3 suppressed their proliferative, migratory and invasive abilities in vitro, and delayed G1 to S phase progression through the cell cycle. Consistent with this, TADA3 silencing increased expression of the epithelial marker E-cadherin and reduced expression of the mesenchymal markers, N-cadherin, Vimentin, Snail, and Slug. To verify the effect of TADA3 on tumor formation and growth in vivo, a mouse tumor xenograft model was established. TADA3 silencing slowed the growth of NSCLC tumor xenografts in nude mice, and excised tumors showed a similarly altered pattern of epithelial-mesenchymal transition (EMT) marker expression. The present results demonstrated the significance of TADA3 in regulating the growth and metastasis of NSCLC and may provide a theoretical basis for early diagnosis and targeted therapy of NSCLC.

Comparison of the long-term outcomes of patients with hepatocellular carcinoma within the Milan criteria treated by ablation, resection, or transplantation.

BACKGROUND: Liver transplantation (LT), resection (LR), and ablation (LA) are three curative-intent treatment options for patients with early hepatocellular carcinoma (HCC). We aimed to develop a prognostic calculator to compare the long-term outcomes following each of these therapies. METHODS: A total of 976 patients with HCC within the Milan criteria who underwent LT, LR, and LA between 2009 and 2019 from four institutions were evaluated. Multistate competing risks prediction models for recurrence-free survival (RFS), recurrence within the Milan criteria (RWM), and HCC-specific survival (HSS) were derived to develop a prognostic calculator. RESULTS: During a median follow-up of 51 months, 420 (43%) patients developed recurrence. In the multivariate analysis, larger tumor size, multinodularity, older age, male, higher alpha-fetoprotein (AFP), higher albumin-bilirubin (ALBI) grade, and the presence of portal hypertension were significantly associated with higher recurrence and decreased survival rates. The RFS and HSS were both significantly higher among patients treated by LT than by LR or LA and significantly higher between patients treated by LR than by LA (all p < 0.001). For multinodular HCC ≤3 cm, although LT had better RFS and HSS than LR or LA, LA was noninferior to LR. An online prognostic calculator was then developed based on the preoperative clinical factors that were independently associated with outcomes to evaluate RFS, RWM, and HSS at different time intervals for all three treatment options. CONCLUSIONS: Although LT resulted in the best recurrence and survival outcomes, LR and LA also offered durable long-term alternatives. This prognostic calculator is a useful tool for clinicians to guide an informed and personalized discussion with patients based on their tumor biology and liver function.

A Novel Nomogram Model to Predict the Recurrence-Free Survival and Overall Survival of Hepatocellular Carcinoma.

Background: Treatments for patients with early-stage hepatocellular carcinoma (HCC) include liver transplantation (LT), liver resection (LR), radiofrequency ablation (RFA), and microwave ablation (MWA), are critical for their long-term survival. However, a computational model predicting treatment-independent prognosis of patients with HCC, such as overall survival (OS) and recurrence-free survival (RFS), is yet to be developed, to our best knowledge. The goal of this study is to identify prognostic factors associated with OS and RFS in patients with HCC and develop nomograms to predict them, respectively. Methods: We retrospectively retrieved 730 patients with HCC from three hospitals in China and followed them up for 3 and 5 years after invasive treatment. All enrolled patients were randomly divided into the training cohort and the validation cohort with a 7:3 ratio, respectively. Independent prognostic factors associated with OS and RFS were determined by the multivariate Cox regression analysis. Two nomogram prognostic models were built and evaluated by concordance index (C-index), calibration curves, area under the receiver operating characteristics (ROC) curve, time-dependent area under the ROC curve (AUC), the Kaplan-Meier survival curve, and decision curve analyses (DCAs), respectively. Results: Prognostic factors for OS and RFS were identified, and nomograms were successfully built. Calibration discrimination was good for both the OS and RFS nomogram prediction models (C-index: 0.750 and 0.746, respectively). For both nomograms, the AUC demonstrated outstanding predictive performance; the DCA shows that the model has good decision ability; and the calibration curve demonstrated strong predictive power. The nomograms successfully discriminated high-risk and low-risk patients with HCC associated with OS and RFS. Conclusions: We developed nomogram survival prediction models to predict the prognosis of HCC after invasive treatment with acceptable accuracies in both training and independent testing cohorts. The models may have clinical values in guiding the selection of clinical treatment strategies.

Microwave irradiation coupled with zero-valent iron that enhances the composite geopolymerization of chromite ore processing residue and its mechanisms.

In this work, microwave (MW) irradiation was employed to enhance the zero-valent iron (ZVI)-dominated de-contamination of chromite ore processing residue (COPR). A coupling system and the traditional two-step procedure were both conducted to evaluate the effects of MW irradiation on the reduction and the incorporation of COPR into the composite materials-based geopolymers. The factors including the ratios of liquid to solid, the mass ratios of ZVI to COPR, and the acid dosage had some obvious influence on the reduction of COPR in the MW system. The compressive strengths of 31.54 and 41.56 MPa were determined from the two-step procedure and the coupling system at the COPR dosage of 10% (mass ratio), respectively. The employment of MW irradiation not only strengthened the formation of the geopolymer matrices but also improved the chemical stabilization of Cr species in the solidified blocks. The coupled process was more conducive to incorporating the treated COPR into the geopolymer-based crystalline microstructures compared with the subsequent usage of ZVI reduction and MW irradiation.

Metabolite secretions of Lactobacillus plantarum YYC-3 may inhibit colon cancer cell metastasis by suppressing the VEGF-MMP2/9 signaling pathway.

BACKGROUND: Colorectal cancer (CRC) is a major clinical challenge, and the gut microbiome plays important roles in the occurrence and metastasis of CRC. Lactobacillus and their metabolites are thought to be able to suppress the growth of CRC cells. However, the antimetastatic mechanism of Lactobacillus or their metabolites toward CRC cells is not clear. Therefore, the aim of this study was to assess the inhibitory mechanism of cell-free supernatants (CFSs) of L. rhamnosus GG, L. casei M3, and L. plantarum YYC-3 on metastasis of CRC cells. RESULTS: YYC-3 CFS showed the highest inhibitory effect on CRC cell growth, invasion and migration, and inhibited MMP2, MMP9, and VEGFA gene and protein expression, and protein secretion. Furthermore, it suppressed the activities of MMPs by gelatin zymography. Moreover, the effective compounds in these CFSs were analyzed by Q Exactive Focus liquid chromatography-mass spectrometry. CONCLUSIONS: Our results showed that metabolite secretions of YYC-3 may inhibited cell metastasis by downregulating the VEGF/MMPs signaling pathway. These data suggest that treatment of CRC cells with metabolites from L. plantarum YYC-3 may reduce colon cancer metastasis.

Generation and characterization of dipeptidyl peptidase-IV inhibitory peptides from trypsin-hydrolyzed α-lactalbumin-rich whey proteins.

Dipeptidyl peptidase-IV (DPP-IV) is an enzyme that break down the antidiabetic hormone glucagon-like peptide-1. Therefore, inhibition of DPP-IV could be an effective strategy to treat Type 2 diabetes (T2D). The α-lactalbumin-rich whey protein concentrate was hydrolyzed by trypsin, and the hydrolysates were then fractionated at a semi-preparative scale using a Superdex Gel filtration Chromatography. The peptides were analyzed by using HPLC coupled with tandem mass spectrometry (RP-HPLC-MS/MS), and their Dipeptidyl peptidase-IV inhibitory activity was determined by the enzymatic assay. Among tested fragments, a potent fragment (LDQWLCEKL), with the half-maximal inhibitory concentration (IC50) of 131 µM was obtained. Further analysis shows that the LDQWLCEKL peptide corresponds to the amino acid sequence of f(115-123) in α-lactalbumin. Furthermore, LDQWLCEKL exhibited a typical non-competitive mode of inhibition. The results indicate that α-lactalbumin contains active peptides with DPP-IV inhibitory activity that may be used to prevent and treat T2D.

miR-148a suppresses autophagy by down-regulation of IL-6/STAT3 signaling in cerulein-induced acute pancreatitis.

Acute pancreatitis (AP) is a type of sterile inflammation of the pancreas, potentially leading to systemic inflammatory response syndrome or multiple organ failure. An emerging evidence that dysfunction of miRNA expression may alter pivotal physiological functions and lead to inflammation infiltration and complication of multiple diseases, including AP. Here, the AP model was successfully replicated using cerulein in vitro and in vivo. RT-qPCR was used to detect low expression of miR-148a in AP. This study verified that IL-6 was a direct target of miR-148a. Over-expression of miR-148a decreased the mRNA and protein levels of IL-6 by RT-qPCR and Elisa. Moreover, over-expression of miR-148a improved the pathological state of AP through H&E and MPO staining and transmission electron microscopy. After over-expressing miR-148a, Western blot and immunohistochemical method were used to confirm the reduction of autophagosomes and autolysosomes, blockade of the levels of p-STAT3, LC3-II, ATG7, ATG4c, Beclin1 and the increased p62 expression in AP. The expression of LAMP-2 was not significantly different. In addition, IL-6 and AG490, the IL-6/STAT3 signaling inhibitor, were used to verify the role of IL-6/STAT3 signaling in the regulation of miR-148a on autophagy in cerulein-induced AP in vitro and in vivo. Taken together, our findings indicate that miR-148a suppresses autophagy via regulating IL-6/STAT3 signaling in cerulein-induced AP in vitro and in vivo. The miR-148a appears to be a promising candidate for the gene therapy of AP.

The Value of SPECT/CT in Monitoring Prefabricated Tissue-Engineered Bone and Orthotopic rhBMP-2 Implants for Mandibular Reconstruction.

Bone tissue engineering shows good prospects for mandibular reconstruction. In recent studies, prefabricated tissue-engineered bone (PTEB) by recombinant human bone morphogenetic proteins (rhBMPs) applied in vivo has found to be an effective alternative for autologous bone grafts. However, the optimal time to transfer PTEB for mandibular reconstruction is still not elucidated. Thus, here in an animal experiment of rhesus monkey, the suitable transferring time for PTEB to reconstruct mandibular defects was evaluated by 99mTc-MDP SPECT/CT, and its value in monitoring orthotopic rhBMP-2 implants for mandibular reconstruction was also evaluated. The result of SPECT/CT showed higher 99mTc-MDP uptake, indicating osteoinductivity, in rhBMP-2 incorporated demineralized freeze-dried bone allograft (DFDBA) and coralline hydroxyapatite (CHA) implants than those without BMP stimulation. 99mTc-MDP uptake of rhBMP-2 implant peaked at 8 weeks following implantation while CT showed the density of these implants increased after 13 weeks' prefabrication. Histology confirmed that mandibular defects were repaired successfully with PTEB or orthotopically rhBMP-2 incorporated CHA implants, in accordance with SPECT/CT findings. Collectively, data shows 99mTc-MDP SPECT/CT is a sensitive and noninvasive tool to monitor osteoinductivity and bone regeneration of PTEB and orthotopic implants. The PTEB achieved peak osteoinductivity and bone density at 8 to 13 weeks following ectopic implantation, which would serve as a recommendable time frame for its transfer to mandibular reconstruction.

[Epidemic Status of Echinococcosis in Gannan Tibetan Autonomous Prefecture of Gansu Province during 2007-2011].

OBJECTIVE: To investigate the prevalence of echinococcosis in Gannan Tibetan Autonomous Prefecture of Gansu Province since the implementation of the echinococcosis control project from Sepical Funds for Central Government Subsidies to Local Public Health (2007-2011). METHODS: Eight counties of Hezuo, Lintan, Zhuoni, Luqu, Maqu, Xiahe, Zhouqu, and Diebu were selected as survey sites. The prevalence in the sampled population was investigated by B ultrasound examination. Hydatid infection in children below 12 years old was serologically investigated by ELISA. The fecal samples from dogs were determined for Echinococcus infection by double antibody sandwich ELISA method. Livestock were dissected through slaughterhouse for pathological examination. Data of echinococcosis cases of Gannan Tibetan Autonomous Prefecture from 2007 to 2011 were collected from the National Infectious Diseases Reporting System, and statistically analyzed by using SPSS 10.0 and Epi info software. RESULTS: A total of 257 823 people received type B ultrasound examination in the 5 years. Five hundred eighty-one echinococcosis cases were found with an overall prevalence of 0.2%, including 578 cases of echinococcosis granulosus and 3 cases of echinococcosis multilocularis. The annual prevalence in the population decreased year by year, from 0.4% (97/21 938) in 2007 to 0.1% (68/63 980) in 2011 (P <0.05). Three hundred and six cases were officially reported to the National Infectious Diseases Reporting System during the period. Among those, female patients accounted for 58.2% (178/306) and male patients 41.8% (128/ 306). By occupation, more infection were found in herdsmen (82.0%, 251/306), followed by farmers (7.8%; 24/306). The cases were mainly distributed in the 20-60 year-old age group, with the highest prevalence in the group of 30-39 years (23.5%, 72/306). The sero-positive rate in children was 4.7% (1 571/33 613), which was highest in 2008 (8.4%, 413/4907), and lowest in 2010 (3.2%, 223/7 021) (P < 0.05). The mean positive rate of coproantigen in dogs was 6.3% (2 511/40 179), decreased from 11.9% (335/2 819) in 2007 to 6.3% (734/11 666) in 2011 (P < 0.05), with lowest positive rate in 2009 (3.7%, 354/9 550). The mean prevalence of livestock was 4.1% (914/22 087), decreased from 8.8% (235/2 658) in 2007 to 2.0% (144/7 347) in 2011 (P < 0.05). CONCLUSION: Since the project implementation for echinococcosis control in 2007, the prevalence of hydatid desease in the population, the sero-positive rate in children, the positive rate of dog coproantigen, and the prevalence in livestock have been significantly decreased.

Salvianolic acid B protects from pulmonary microcirculation disturbance induced by lipopolysaccharide in rat.

The aim of the present study was to examine the effect and possible mechanism of salvianolic acid B (SalB) on pulmonary microcirculation disturbance induced by lipopolysaccharide (LPS) in rat. Male Sprague-Dawley rats were subjected to thoracotomy under continuous anesthesia and mechanical ventilation. Albumin leakage from pulmonary capillary and the numbers of leukocytes adherent to the pulmonary capillary wall were determined for 60 min by an upright microscope upon LPS (2 mg · kg(-1) · h(-1)) infusion with or without administration of SalB (5 mg · kg(-1) · h(-1)). Pulmonary tissue wet-to-dry weight ratio, tumor necrosis factor α, and interleukin 8 in plasma and bronchoalveolar lavage fluid were measured. In addition, the expressions of E-selectin, intercellular adhesion molecule 1, and myeloperoxidase in pulmonary tissue were assessed by immunohistochemistry. The expressions of aquaporin 1 (AQP-1), AQP-5, metalloproteinase 2 (MMP-2), and MMP-9 were assessed by Western blot assay. Pretreatment with SalB significantly attenuated LPS-induced pulmonary microcirculatory disturbance, including the increase in leukocyte adhesion and albumin leakage. In addition, LPS increased pulmonary tissue wet-to-dry weight ratio and tumor necrosis factor α and interleukin 8 levels in plasma and bronchoalveolar lavage fluid enhanced the expression of E-selectin, intercellular adhesion molecule 1, myeloperoxidase, MMP-2, and MMP-9, whereas it decreased the expression of AQP-1 and AQP-5 in pulmonary tissue, all of which were attenuated by SalB pretreatment. Salvianolic acid B pretreatment improves pulmonary microcirculation disturbance and lung injury on LPS exposure. More studies are required to evaluate the potential of SalB as an option for protecting lung from endotoxemia.

[Effect of Tongfu granules and its constituents on barrier function of small intestine in rats with sepsis].

OBJECTIVE: To investigate the effects of Tongfu granules and its constituents on barrier function of small intestine in rats with sepsis. METHODS: The male rats were divided into model group, Tongfu granules group, Rhubarb group and Magnoliae cortex group by random digits table, normal rats as control group. Intraperitoneal injection of lipopolysaccharide (LPS, 6 mg/kg) was used to reproduce sepsis model. After establishment of model, rats in Tongfu granules group were given Tongfu granules 28 g×kg(-1)×d(-1) by gavage, and Rhubarb group and Magnoliae cortex group rats were given Rhubarb or Magnoliae cortex 5 g×kg(-1)×d(-1) by gavage, while the model group was given normal saline in same quantity, once a day. Blood samples of rats were collected at 24, 48, 72 hours for measuring tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) with enzyme linked immunosorbent assay (ELISA). The pathological changes in intestinal mucosa were observed, and the pathological scores was estimated at 72 hours. RESULTS: The levels of TNF-α and IL-8 were significantly higher in model group than those in control group at different time points. The serum levels of TNF-α and IL-8 were significantly lower in treatment groups than those in model group, and the level of TNF-α (ng/L) in Tongfu granules group was significantly lower than that in Rhubarb and Magnoliae cortex groups at different time points (24 hours: 44.64±1.48 vs. 47.18±1.83 and 46.96±2.23, 48 hours: 51.38±1.36 vs. 57.17±2.23 and 59.41±2.01, 72 hours: 55.54±2.58 vs. 64.34±1.02 and 65.96±3.45, all P<0.05), and IL-8 (ng/L) level at 72 hours was significantly lower than that in Magnoliae cortex group (65.53±4.52 vs. 69.14±2.82,P<0.05). The scores of the lesions were significantly higher in model group than that in control group (3.90±0.17 vs. 0). The scores of Rhubarb group, Magnoliae cortex group and Tongfu granules group were 3.15±0.28, 3.18±0.08, and 2.95±0.15, respectively, which were lower than those of the model group (all P<0.01), and the Tongfu granules group descended obviously than other groups. In control group, the intercellular tight junctions were normal, and the morphology of microvilli and mitochondria was also normal. In model group, the microvilli of intestinal mucosa of the small intestine were absent or disintegrated. The intercellular tight junctions were seen to be blurred in Rhubarb and Magnoliae cortex groups, and they were close to normal state in Tongfu granules group. Their integrity was better preserved compared with that of the model group. CONCLUSION: Injury of barrier function of the small intestine was found in septic rat. It was found that traditional Chinese medicine Tongfu granules, Rhubarb and Magnoliae cortex could protect the barrier function of the small intestine by decreasing the TNF-α and IL-8 levels in septic rats. Above-mentioned effects of Tongfu granules were better than Rhubarb and Magnoliae cortex.

[A clinical study on the diagnostic criteria of multiple organ dysfunction syndrome].

OBJECTIVE: To develop diagnostic criteria of multiple organ dysfunction syndrome (MODS) by a prospective and multi-center clinical investigation. METHODS: The data of 1087 MODS cases obtained from ICU of 37 hospitals from March 2002 to January 2005 in 11 provinces in China were analyzed in order to derive the diagnostic criteria of MODS. RESULTS: This MODS diagnostic criteria involved 7 organs. To diagnose MODS, the original cause of MODS should be identified, then there should be two or more organs showing signs of dysfunction. The criteria for organ dysfunction were as follows. (1) Cardiovascular system: SBP < 90 mm Hg (1 mm Hg = 0.133 kPa), MAP < 70 mm Hg, signs of shock, ventricular tachycardia, ventricular fibrillation, or myocardial infarction; (2) Respiratory system: oxygenation index < 300 mm Hg; (3) Nervous system: indifference, restlessness, lethargy, light coma, or deep coma, Glasgow score < or = 14; (4) Blood system: PLT < 100 x 10(9)/L; CT, APTT, and PT prolonged or shortened; positive plasma protamine paracoagulation; (5) Liver: TBIL > 20. 5 micromol/L, ALB < 28 g/L; (6) Kidney: Cr > 123.8 micromol/L, urinary volume < 500 ml/24 h; (7) Gastro-intestine: bowel sounds decreased or disappeared; retention in the stomach, or positive occult blood feces with dark stools or haematemesis; intraabdominal pressure (intravesical pressure) > or = 11 cm H2O (1 cm H2O = 0.098 kPa). Any organ function met with one of the above conditions was considered to have dysfunction. CONCLUSIONS: This diagnostic criterion of multiple organ dysfunction syndrome has been developed by this research, but it needs to accumulate experience by clinical practice and to revise the diagnosis criteria.

Response of mesenchymal stem cells to shear stress in tissue-engineered vascular grafts.

AIM: Recent studies have demonstrated that mesenchymal stem cells (MSCs) can differentiate into endothelial cells. The effect of shear stress on MSC differentiation is incompletely understood, and most studies have been based on two-dimensional systems. We used a model of tissue-engineered vascular grafts (TEVGs) to investigate the effects of shear stress on MSC differentiation. METHODS: MSCs were isolated from canine bone marrow. The TEVG was constructed by seeding MSCs onto poly-epsilon-caprolactone and lactic acid (PCLA) scaffolds and subjecting them to shear stress provided by a pulsatile bioreactor for four days (two days at 1 dyne/cm(2) to 15 dyne/cm(2) and two days at 15 dyne/cm(2)). RESULTS: Shear stress significantly increased the expression of endothelial cell markers, such as platelet-endothelial cell adhesion molecule-1 (PECAM-1), VE-cadherin, and CD34, at both the mRNA and protein levels as compared with static control cells. Protein levels of alpha-smooth muscle actin (alpha-SMA) and calponin were substantially reduced in shear stress-cultured cells. There was no significant change in the expression of alpha-SMA, smooth muscle myosin heavy chain (SMMHC) or calponin at the mRNA level. CONCLUSION: Shear stress upregulated the expression of endothelial cell-related markers and downregulated smooth muscle-related markers in canine MSCs. This study may serve as a basis for further investigation of the effects of shear stress on MSC differentiation in TEVGs.

[Tissue engineering of vascular graft from decellularized arterial matrix and mesenchymal stem cells.].

OBJECTIVE: To investigate the method of constructing small-diameter vascular grafts from xenogenic decellularized arterial matrices and mesenchymal stem cells (MSCs). METHODS: Porcine iliac arteries were decellularized by detergent and trypsin treatment. Histology, mechanical strength and porosity experiments were performed to evaluate the properties of decellularized matrices. MSCs were isolated from bone marrow of dogs and expanded ex vivo. Decellularized matrices were seeded with MSCs and further cultured in a pulsatile bioreactor. Morphological features of the tissue engineered grafts were assayed by HE staining and scanning electron microscopy. RESULTS: After cell extraction, absence of cellular components and preservation of extracellular matrix were verified. Mechanical strength of decellularized matrices was slightly reduced compared with native arteries. Porosity of decellularized matrices was 94.9%. Decellularized matrices were successfully seeded with MSCs, which grew to a near-confluent monolayer under flow conditions and MSCs were highly elongated and oriented to the flow direction. CONCLUSION: Small-diameter vascular grafts can be constructed by seeding MSCs onto xenogenic decellularized arterial matrices and culturing in a pulsatile bioreactor.

Magnolol attenuates sepsis-induced gastrointestinal dysmotility in rats by modulating inflammatory mediators.

AIM: To investigate the protective effects of magnolol on sepsis-induced inflammation and intestinal dysmotility. METHODS: Sepsis was induced by a single intraperitoneal injection of lipopolysaccharide (LPS). Male Wistar rats were randomly assigned to one of three treatment groups: magnolol prior to LPS injection (LPS/Mag group); vehicle prior to LPS injection (LPS/Veh group); vehicle prior to injection of saline (Control/Veh). Intestinal transit and circular muscle mechanical activity were assessed 12 h after LPS injection. Tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), monocyte chemoattractant protein-1 (MCP-1) and inducible nitric oxide synthase (iNOS) mRNA in rat ileum were studied by RT-PCR 2 h after LPS injection. Nuclear factor-kappaB (NF-kappaB) activity in the intestine was also investigated at this time using electrophoretic mobility shift assay. In addition, antioxidant activity was determined by measuring malondialdehyde (MDA) concentration and superoxide dismutase (SOD) activity in the intestine 2 h after LPS injection. RESULTS: Magnolol significantly increased intestinal transit and circular muscle mechanical activity in LPS-treated animals. TNF-alpha, MCP-1 and iNOS mRNA expression in the small intestine were significantly reduced after magnolol treatment in LPS-induced septic animals, compared with untreated septic animals. Additionally, magnolol significantly increased IL-10 mRNA expression in septic rat ileum. Magnolol also significantly suppressed NF-kappaB activity in septic rat intestine. In addition, magnolol significantly decreased MDA concentration and increased SOD activity in rat ileum. CONCLUSION: Magnolol prevents sepsis-induced suppression of intestinal motility in rats. The potential mechanism of this benefit of magnolol appears to be modulation of self-amplified inflammatory events and block of oxidative stress in the intestine.

[The effect of recombinant interleukin-10/Fc fusion protein on lipopolysaccharide-induced acute lung injury in mice].

OBJECTIVE: To clarify the regulatory role and mechanism of recombinant interleukin-10/Fc (rIL-10/Fc) fusion protein on inflammatory parameters during development of acute lung injury (ALI) induced by lipopolysaccharide (LPS) in a murine model. METHODS: An ALI model was reproduced by intra-tracheal injection of LPS. rIL-10/Fc was administered intraperitoneally. One hundred and thirty-two BALB/c mice were divided into four groups, including saline control group, rIL-10/Fc control group, ALI model group, and rIL-10/Fc treatment group. Twenty-four-hour survival rate was determined in 25 mice of each group. The number of inflammatory cells and inflammatory mediators in bronchial-alveolar lavage fluid (BALF), tumor necrosis factor-alpha (TNF-alpha) and IL-1 beta, and also lung myeloperoxidase (MPO) activity, lung wet/dry (W/D) ratio were determined in the rest of mice. Pathological changes in lung were examined with hematoxylin-eosin (HE) staining, and inflammatory change was evaluated under microscope. RESULTS: Levels of TNF-alpha and IL-1 beta in BALF were substantially increased 4 hours after intra-tracheal LPS (both P<0.01), and they were lowered but without significant difference after rIL-10/Fc administration. However, rIL-10/Fc fusion protein markedly attenuated release of TNF-alpha at 8 hours and 12 hours, and IL-1 beta was lowered at 12 hours after LPS challenge. Pre-treatment with rIL-10/Fc fusion protein significantly improved survival rate at 24 hours in LPS challenged mice (P<0.01). There was no significant difference in cell count in BALF, MPO, lung W/D ratio, after treatment of rIL-10/Fc fusion protein. Obvious inflammatory changes were found in lung was found pathologically at 24 hours after LPS injection, but there was no significant difference compared with ALI mice with rIL-10/Fc fusion protein administration. CONCLUSION: rIL-10/Fc fusion protein inhibits release of TNF-alpha and IL-1 beta in BALF in a LPS-induced ALI murine model. rIL-10/Fc fusion protein improves survival rate in ALI mice by decreasing the release of pro-inflammatory cytokines.

Transplantation of autologous bone marrow mononuclear cells for patients with lower limb ischemia.

BACKGROUND: Many treatment options for lower limb ischemia are difficult to apply for the patients with poor arterial outflow or with poor general conditions. The effect of medical treatment alone is far from ideal, especially in patients with diabetic foot. A high level amputation is inevitable in these patients. This study aimed to explore the effect of transplantation of autologous bone marrow mononuclear cells on the treatment of lower limb ischemia and to compare the effect of intra-arterial transplantation with that of intra-muscular transplantation. METHODS: In this clinical trial, 32 patients with lower limb ischemia were divided into two groups. Group 1 (16 patients with 18 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-muscular injection into the affected limbs; and group 2 (16 patients with 17 affected limbs) received transplantation of autologous bone marrow mononuclear cells by intra-arterial injection into the affected limbs. Rest pain, coldness, ankle/brachial index (ABI), claudication, transcutaneous oxygen pressure (tcPO(2)) and angiography (15 limbs of 14 patients) were evaluated before and after the mononuclear cell transplantation to determine the effect of the treatment. RESULTS: Two patients died from heart failure. The improvement of rest pain was seen in 76.5% (13/17) of group 1 and 93.3% (14/15) of group 2. The improvement of coldness was 100% in both groups. The increase of ABI was 44.4% (8/18) in group 1 and 41.2% (7/17) in group 2. The value of tcPO(2) increased to 20 mmHg or more in 20 limbs. Nine of 15 limbs which underwent angiography showed rich collaterals. Limb salvage rate was 83.3% (15/18) in group 1 and 94.1% (16/17) in group 2. There was no statistically significant difference in the effectiveness of the treatment between the two groups. CONCLUSIONS: Transplantation of autologous bone marrow mononuclear cells is a simple, safe and effective method for the treatment of lower limb ischemia, and the two approaches for the implantation, intra-muscular injection and intra-arterial injection, show similar results.

[Effect of Qishen Huoxue Granule in treating severe sepsis].

OBJECTIVE: To study the anti-inflammatory effect and the therapeutic efficacy of Qishen Huoxue Granule (QHG) in treating severe sepsis. METHODS: One hundred and sixty-seven patients with severe sepsis were enrolled and randomly assigned to two groups, the 85 patients in the control group treated with conventional Western medicine and the 82 in the QHG group treated with conventional Western medicine plus QHG. Changes of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-10 (IL-10), procalcitonin (PCT), Marshall score, APACHE II score, ICU stay time and 28-day mortality were monitored and compared. RESULTS: Compared with the control group, IL-6 and TNF-alpha levels, the ICU stay time and 28-day mortality were significantly lower in the QHG group (all P < 0.05). During the QHG treatment, no severe adverse event was observed. CONCLUSION: The integrative treatment could reduce the blood levels of IL-6 and TNF-alpha, shorten the ICU stay time and decrease the 28-day mortality of patients with sepsis, showing a favor therapeutic prospect.

[Liver injury and intervention of compound 912 liquid on it in rats with endotoxemia].

OBJECTIVE: To investigate the liver injury in model rats with endotoxemia and to observe the protective effect of Compound 912 Liquid on it. METHODS: Rats were randomly divided into three groups, the endotoxemia model group (EMG, injected by lipoplysaccharides (LPS) peritoneally), the intervention group (IG, treated with Compound 912 Liquid via gastrogavage 1 h before model establishing) and the normal control group (NCG). Blood samples of rats were taken at the time points of the 2nd, 4th, 8th, 12th, 48th, 72nd hour and the 7th day after modeling for measuring liver function, levels of plasmatic endotoxin, tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10). The pathological change of liver was observed using light microscope and electro-transmission microscope. RESULTS: The peak concentration of endotoxin detected at 2 hour after modeling in the IG was significantly lower than that in the EMG (0.358 +/- 0.056 vs 0.685 +/- 0.030), but insignificant difference (P > 0.05) was shown between them in TNF-alpha level. The level of IL-10 continuously rose in IG after treatment, it was still higher than normal level until day 7 (49.096 +/- 4.076 vs 43.454 +/- 5.928, P < 0.05). CONCLUSION: LPS can induce the increase of serum inflammatory cytokines and anti-inflammatory cytokines in rats to injure liver. Therefore, the inflammatory reaction indicated by LPS may be one of the mechanisms for liver injury. Preventive medication with Compound 912 Liquid showed a significant liver protective effect.