Si-Wu-Tang attenuates liver fibrosis via regulating lncRNA H19-dependent pathways involving cytoskeleton remodeling and ECM deposition / 中国天然药物

Liver fibrosis is a dynamic wound-healing response characterized by the agglutination of the extracellular matrix (ECM). Si-Wu-Tang (SWT), a traditional Chinese medicine (TCM) formula, is known for treating gynecological diseases and liver fibrosis. Our previous studies demonstrated that long non-coding RNA H19 (H19) was markedly upregulated in fibrotic livers while its deficiency markedly reversed fibrogenesis. However, the mechanisms by which SWT influences H19 remain unclear. Thus, we established a bile duct ligation (BDL)-induced liver fibrosis model to evaluate the hepatoprotective effects of SWT on various cells in the liver. Our results showed that SWT markedly improved ECM deposition and bile duct reactions in the liver. Notably, SWT relieved liver fibrosis by regulating the transcription of genes involved in the cytoskeleton remodeling, primarily in hepatic stellate cells (HSCs), and influencing cytoskeleton-related angiogenesis and hepatocellular injury. This modulation collectively led to reduced ECM deposition. Through extensive bioinformatics analyses, we determined that H19 acted as a miRNA sponge and mainly inhibited miR-200, miR-211, and let7b, thereby regulating the above cellular regulatory pathways. Meanwhile, SWT reversed H19-related miRNAs and signaling pathways, diminishing ECM deposition and liver fibrosis. However, these protective effects of SWT were diminished with the overexpression of H19 in vivo. In conclusion, our study elucidates the underlying mechanisms of SWT from the perspective of H19-related signal networks and proposes a potential SWT-based therapeutic strategy for the treatment of liver fibrosis.

Cell-derived nanovesicles from mesenchymal stem cells as extracellular vesicle-mimetics in wound healing

Wound healing is a dynamic process that involves a series of molecular and cellular events aimed at replacing devitalized and missing cellular components and/or tissue layers. Recently, extracellular vesicles (EVs), naturally cell-secreted lipid membrane-bound vesicles laden with biological cargos including proteins, lipids, and nucleic acids, have drawn wide attention due to their ability to promote wound healing and tissue regeneration. However, current exploitation of EVs as therapeutic agents is limited by their low isolation yields and tedious isolation processes. To circumvent these challenges, bioinspired cell-derived nanovesicles (CDNs) that mimic EVs were obtained by shearing mesenchymal stem cells (MSCs) through membranes with different pore sizes. Physical characterisations and high-throughput proteomics confirmed that MSC-CDNs mimicked MSC-EVs. Moreover, these MSC-CDNs were efficiently uptaken by human dermal fibroblasts and demonstrated a dose-dependent activation of MAPK signalling pathway, resulting in enhancement of cell proliferation, cell migration, secretion of growth factors and extracellular matrix proteins, which all promoted tissue regeneration. Of note, MSC-CDNs enhanced angiogenesis in human dermal microvascular endothelial cells in a 3D PEG-fibrin scaffold and animal model, accelerating wound healing in vitro and in vivo. These findings suggest that MSC-CDNs could replace both whole cells and EVs in promoting wound healing and tissue regeneration.

Remodeling of the liver fibrosis microenvironment based on nilotinib-loaded multicatalytic nanozymes with boosted antifibrogenic activity

Liver fibrosis is a reversible pathological process caused by chronic liver damage and a major risk factor for hepatocellular carcinoma (HCC). Hepatic stellate cell (HSC) activation is considered the main target for liver fibrosis therapy. However, the efficiency of this strategy is limited due to the complex microenvironment of liver fibrosis, including excessive extracellular matrix (ECM) deposition and hypoxia-induced imbalanced ECM metabolism. Herein, nilotinib (NIL)-loaded hyaluronic acid (HA)-coated Ag@Pt nanotriangular nanozymes (APNH NTs) were developed to inhibit HSCs activation and remodel the microenvironment of liver fibrosis. APNH NTs efficiently eliminated intrahepatic reactive oxygen species (ROS) due to their inherent superoxide dismutase (SOD) and catalase (CAT) activities, thereby downregulating the expression of NADPH oxidase-4 (NOX-4) and inhibiting HSCs activation. Simultaneously, the oxygen produced by the APNH NTs further alleviated the hypoxic microenvironment. Importantly, the released NIL promoted collagen depletion by suppressing the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1), thus synergistically remodeling the microenvironment of liver fibrosis. Notably, an in vivo study in CCl4-induced mice revealed that APNH NTs exhibited significant antifibrogenic effects without obvious long-term toxicity. Taken together, the data from this work suggest that treatment with the synthesized APNH NTs provides an enlightening strategy for remodeling the microenvironment of liver fibrosis with boosted antifibrogenic activity.

Cytokine receptor-like factor 1 (CRLF1) promotes cardiac fibrosis via ERK1/2 signaling pathway / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease. Anti-fibrosis treatment is a significant therapy for heart disease, but there is still no thorough understanding of fibrotic mechanisms. This study was carried out to ascertain the functions of cytokine receptor-like factor 1 (CRLF1) in cardiac fibrosis and clarify its regulatory mechanisms. We found that CRLF1 was expressed predominantly in cardiac fibroblasts. Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction, but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-‍β1 (TGF‍-‍β1). Gain- and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts (NMCFs) with or without TGF-‍β1 stimulation. CRLF1 overexpression increased cell viability, collagen production, cell proliferation capacity, and myofibroblast transformation of NMCFs with or without TGF‍-‍β1 stimulation, while silencing of CRLF1 had the opposite effects. An inhibitor of the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and different inhibitors of TGF-‍β1 signaling cascades, comprising mothers against decapentaplegic homolog (SMAD)‍-dependent and SMAD-independent pathways, were applied to investigate the mechanisms involved. CRLF1 exerted its functions by activating the ERK1/2 signaling pathway. Furthermore, the SMAD-dependent pathway, not the SMAD-independent pathway, was responsible for CRLF1 up-regulation in NMCFs treated with TGF-‍β1. In summary, activation of the TGF-‍β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression. CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway. CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.

Mechanism of TRPC in promoting extracellular matrix deposition of glomerular mesangial cells / 西安交通大学学报(医学版)

【Objective】 To explore the role and mechanism of TRPC in promoting extracellular matrix (ECM) deposition in rat glomerular mesangial cells (HBZY-1). Methods Immunofluorescence staining was performed to observe the distribution and expression of TRPC1 and TRPC6 in HBZY-1 cells. After AngⅡ stimulation, qRT-PCR and Western blotting were used to detect the mRNA and protein expressions of Gαq/PLCβ4/TRPC signaling pathway main proteins and ECM deposition indicators (α-SMA, collagenⅢ and fibronectin). By silencing the expressions of TRPC1 and TRPC6 by RNA interference, the expressions of ECM deposition indicators were detected. Changes in [Ca2+]i influx were determined through Fluo-4AM Ca2+ imaging. 【Results】 Both TRPC1 and TRPC6 were expressed in HBZY-1, and were mainly located in cell membrane and cytoplasm. After AngⅡ stimulation, Gαq/PLCβ4/TRPC signaling pathway was activated, and the mRNA and protein expressions of Gαq, PLCβ4, TRPC1 and TRPC6 were all increased (P<0.05). [Ca2+]i influx also increased (P<0.01), and the mRNA and protein expressions of ECM deposition indicators (α-SMA, ColⅢ and Fn) were upregulated (P<0.05). Silencing the expressions of TRPC1 and TRPC6 by RNA interference led to decreased [Ca2+]i influx (P<0.05), and downregulated mRNA and protein expressions of ECM deposition indicators in HBZY-1 cells (P<0.05). The results suggested that inhibition of TRPC expressions could inhibit AngⅡ induced ECM deposition in HBZY-1 cells, which might be associated with decreased [Ca2+]i influx. 【Conclusion】 TRPC may be a novel therapeutic target of renal fibrosis.

Qianjin Wenwu decoction suppresses renal interstitial fibrosis by enhancing the degradation of extracellular matrix in mice with unilateral ureteral obstruction / 中国天然药物

Diabetic kidney disease (DKD) is the most common complication of diabetes mellitus (DM). Qianjin Wenwu decoction (QWD), a well-known traditional Korean medicine, has been used for the treatment of DKD, with satisfactory therapeutic effects. This study was designed to investigate the active components and mechanisms of action of QWD in the treatment of DKD. The results demonstrated that a total of 13 active components in five types were found in QWD, including flavonoids, flavonoid glycosides, phenylpropionic acids, saponins, coumarins, and lignins. Two key proteins, TGF-β1 and TIMP-1, were identified as the target proteins through molecular docking. Furthermore, QWD significantly suppressed Scr and BUN levels which increased after unilateral ureteral obstruction (UUO). Hematoxylin & eosin (H&E) and Masson staining results demonstrated that QWD significantly alleviated renal interstitial fibrosis in UUO mice. We also found that QWD promoted ECM degradation by regulating MMP-9/TIMP-1 homeostasis to improve renal tubulointerstitial fibrosis and interfere with the expression and activity of TGF- β1 in DKD treatment. These findings explain the underlying mechanism of QWD for the treatment of DKD, and also provide methodological reference for investigating the mechanism of traditional medicine in the treatment of DKD.

Inhibiting collagen I production and tumor cell colonization in the lung via miR-29a-3p loading of exosome-/liposome-based nanovesicles

The lung is one of the most common sites for cancer metastasis. Collagens in the lung provide a permissive microenvironment that supports the colonization and outgrowth of disseminated tumor cells. Therefore, down-regulating the production of collagens may contribute to the inhibition of lung metastasis. It has been suggested that miR-29 exhibits effective anti-fibrotic activity by negatively regulating the expression of collagens. Indeed, our clinical lung tumor data shows that miR-29a-3p expression negatively correlates with collagen I expression in lung tumors and positively correlates with patients' outcomes. However, suitable carriers need to be selected to deliver this therapeutic miRNA to the lungs. In this study, we found that the chemotherapy drug cisplatin facilitated miR-29a-3p accumulation in the exosomes of lung tumor cells, and this type of exosomes exhibited a specific lung-targeting effect and promising collagen down-regulation. To scale up the preparation and simplify the delivery system, we designed a lung-targeting liposomal nanovesicle (by adjusting the molar ratio of DOTAP/cholesterol-miRNAs to 4:1) to carry miR-29a-3p and mimic the exosomes. This liposomal nanovesicle delivery system significantly down-regulated collagen I secretion by lung fibroblasts in vivo, thus alleviating the establishment of a pro-metastatic environment for circulating lung tumor cells.

The Dual Role of LOXL4 in Pathogenesis and Development of Human Malignant Tumors / 中国生物化学与分子生物学报

Lysyl oxidase like 4 (LOXL4) is one member of the LOX protein family and is a secreted copper-dependent amine oxidase involved in the assembly and maintenance of extracellular matrix (ECM). LOXL4 is up-regulated in human liver cancer, gastric cancer, breast cancer, cervical cancer, head and neck squamous cell carcinoma, esophageal carcinoma and colorectal cancer, but down-regulated in human bladder and lung cancer. It also inhibits tumor growth in bladder and lung cancer, suggesting that L0XL4 has a dual role of promoting or inhibiting tumors in different types of human malignant tumors. L0XL4 in tumor cell exosomes promotes cell matrix adhesion and cell migration by activating the FAK/Src pathway, which is dependent on its amine oxidase activity through a hydrogen peroxide-mediated mechanism. Exosome-mediated L0XL4 can also promote tumor cell proliferation and immune escape by activating the PI3K/Akt signaling pathway. L0XL4 can be transported to macrophages via exosomes in tumor cells, where it further activates the immunosuppression function of cells and the expression of programmed death ligand 1 (PD-L1) via STAT1- and STAT3-mediated signaling pathways. It then will trigger the immunosuppressive function of macrophages and promote the immune escape of tumor cells. In addition, LOXL4 can also exert the tumor suppressive function by activating p53 and inhibiting the Ras/ERK pathway. This paper mainly reviews the structure and the function of LOXL4, and the relationship between LOXL4 and the pathogenesis and development of human malignant tumors. We then further explore the application of LOXL4 in the study of malignant tumors, laying a theoretical foundation for its future utilization in the clinical diagnosis and treatment, and screening of prognostic markers of human malignant tumors.

Contribution and Regulation of Forkhead Transcription Factor O1 during the Renal Fibrosis / 中国生物化学与分子生物学报

Chronic kidney disease (CKD) has become a chronic disease that cannot be ignored all over the world. Renal fibrosis is the final result and key pathological features of CKD. The whole process of renal fibrosis is associated with the repair and healing of kidney damage, and this long-term chronic stimulation promotes the renal fibrosis to the kidney failure, it is characterized by continuous activation of myofibroblasts and excessive production and deposition of extracellular matrix. Renal fibrosis is a synergistic process mediated by a variety of signaling pathways, and TGF-β / Smad and Wnt / β-catenin signaling pathways are two of the most classic ones. FoxO1 is an important transcription factor in the human body, which is expressed in a variety of cell types and plays an important role in a variety of cell life activities. Current researches have shown that FoxO1 plays an important role in renal fibrosis. FoxO1 regulates the occurrence and development of renal fibrosis through various signaling pathways such as STAT, SIRT1, Wnt / β-catenin, PI-3K / Akt. For example, FoxO1 / STAT regulates TIF and renal tubule apoptosis, SIRT1 / FoxO1 regulates oxidative stress response and lipotoxicity, FoxO1 / β-catenin inhibits the Wnt / β-catenin signaling pathway. In addition, some related studies suggest that FoxO1 / Smad may play a role in renal fibrosis. The contribution and regulation of FoxO1 in the renal fibrosis are reviewed here in order to further clarify the pathogenesis of renal fibrosis, which may bring new clues and prospects for the treatment of CKD.

Progress in Biomechanics of Materials in 2021 / 医用生物力学

Tissue has a complex three-dimensional (3D) dynamic structure, and is affected by various forms of forces. Cells sense mechanical forces from extracellular matrix (ECM), and the mechanical micro-environment constructed by ECM regulates different biological functions of cells. To prepare biomaterials which can simulate the ECM mechanical micro-environment of tissues is one of the research hot spots and difficulties in biomechanical field. Different physical and chemical properties of biomaterials endow materials with specific mechanical properties, which further affect the behavior and function of cells. Based on the latest literature of biomechanics of materials in the year 2021, this study mainly focused on the role of novel mechanical biomaterials in regulating cell biological behavior and application in tissue engineering. The future development direction in the field of biomechanics of materials was also discussed.

Influences of Cell-ECM Adhesion on Migration of Tumor Cells Regulated by ECM Stiffness: A Model Study / 医用生物力学

Objective To study the influence of cell-extracellular matrix (ECM) adhesion on migration of tumor cells regulated by ECM stiffness. Methods The cellular Potts model (CPM) was established to simulate tumor cell growth and cellular immune feedback system. The effects from mechanical behavior of cells on cell-ECM adhesion were observed, and the migration of tumor cells under different ECM was analyzed. Results The ECM stiffness could influence the migration rate of tumor cells. The change of ECM stiffness regulated the adhesion force between cells and ECM, and the change of adhesion force would influence the migration rate of cells. Conclusions The migration and distribution patterns of cells are closely related to the adhesion and stiffness of ECM. The increase in ECM stiffness can effectively promote the migration rate of tumor cells, and the further increase in ECM stiffness inhibits the migration of tumor cells. These findings may further reveal dynamic changes of ECM, adhesion and mechanical performance of tumor cell migration.

SMYD3-PARP16 axis accelerates unfolded protein response and mediates neointima formation

Neointimal hyperplasia after vascular injury is a representative complication of restenosis. Endoplasmic reticulum (ER) stress-induced unfolded protein response (UPR) is involved in the pathogenesis of vascular intimal hyperplasia. PARP16, a member of the poly(ADP-ribose) polymerases family, is correlated with the nuclear envelope and the ER. Here, we found that PERK and IRE1

Death and Complexity / Muerte y complejidad / Morte e complexidade

Abstract: There is no such thing as a science of death, although there is a science of life, as it happens. Death is not so much the subject matter of science but an experience, and death experiences we find abundantly in the literature. Now, experience is told not so much in a scientific tenure but as a narrative. Within the framework of bioethics, death comes closer, particularly what is usually known as end-of-life dilemmas, i.e., palliative care, a most sensitive arena, if there is any at all. This paper argues about the interplay or dialogue between death and complexity science. It claims that the knowledge of death is truly the knowledge of life and provides three arguments that lead to the central claim. The first argument is very much close to a kind of heuristic for knowing about death, while the second shows the challenge of knowing death. The third one consists of a reappraisal of death within an extensive cultural or civilizing framework. Lastly, some open-ended conclusions are drawn.

Resumen: No existe cosa tal como una ciencia de la muerte, sin embargo si existe una ciencia de la vida, por suerte. La muerte no se trata de un asunto de ciencia sino de una experiencia, y experiencias de muerte abundan en la literatura. De hecho, la experiencia se cuenta no tanto en un tono científico sino más bien como una narrativa. Dentro del marco de la bioética, la muerte se aborda de una manera más cercana, particularmente en los que se conocen como dilemas del fin de la vida, por ejemplo, el cuidado paliativo, una arena muy sensible, si es que la hay. Este artículo discute sobre la interacción o el diálogo entre la muerte y la ciencia de la complejidad. Afirma que el conocimiento de la muerte en verdad es el conocimiento de la vida y da tres argumentos que llevan a esta afirmación central. El primer argumento es muy cercano a un tipo de heurística por conocer acerca de la muerte, mientras que el segundo expone el reto de conocer la muerte. El tercero consiste en una revaluación de la muerte dentro de un amplio marco cultural o civilizador. Por último, se esbozan algunas conclusiones abiertas.

Resumo: Não existe uma ciência da morte como tal, contudo sim existe uma ciência da vida. A morte não se trata de um assunto de ciência, mas sim de uma experiência, e experiências de morte abundam na literatura. De fato, a experiência se conta não tanto em um tom científico, mas sim como uma narrativa. No âmbito da bioética, a morte é abordada de uma maneira mais íntima, particularmente no que se conhece como "dilemas do fim da vida", por exemplo, no cuidado paliativo, uma arena muito sensível, sim é que a há. Neste artigo, discute-se sobre a interação ou o diálogo entre a morte e a ciência da complexidade. Afirma-se que o conhecimento da morte em verdade é o conhecimento da vida e são apresentados três argumentos que levam a essa afirmação central. O primeiro argumento é muito próximo de um tipo de heurística por conhecer sobre a morte, enquanto o segundo expõe o desafio de conhecer a morte. O terceiro consiste em uma reavaliação da morte dentro de um amplo referencial cultural ou civilizador. Por último, são esboçadas algumas conclusões abertas.

Effect of Different Effective Parts of Taohe Chengqitang on Synthesis and Degradation of Extracellular Matrix in HK-2 Cells Induced by TGF-β1 / 中国实验方剂学杂志

Objective:To explore the effect of different effective parts of Taohe Chengqitang on the synthesis and degradation of extracellular matrix in human kideny-2（HK-2） cells induced by transforming growth factor-β1（TGF-β1）. Method:Petroleum ether extract, ethyl acetate extract, n-butanol extract, raffinate and polysaccharide extract, mirabilite extract were extracted with 70% ethanol by systematic solvent method. The HK-2 cell fibrosis model induced by TGF-β1 was built to intervene the cells in different parts of Taohe Chengqitang with different concentrations (0, 50, 100, 200, 400, 800 mg·L-1). Enzyme-linked immunosorbent assay（ELISA）kit assay was used to detect collagen（Col）-Ⅰα1 and fibronectin （FN）in supernatant to screen out the main active parts. Cell counting kit-8 （CCK-8）method was used to determine the best concentration of intervention site of bioactive components. Western blot analysis was used to detect the expression levels of Col-Ⅰ, Col-Ⅲ, matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase inhibitor2 (TIMP2), and connective tissue growth factor (CTGF). Immunofluorescence assay was used to detect the expression of α-smooth muscle actin（α-SMA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） analysis was used to detect the mRNA expression of plasminogen activator inhibitor-1（PAI-1）. Result:ELISA kit assay demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract significantly reduced the Col-Ⅰα1 and FN content at the concentrations of 200 and 400 mg·L-1 (P<0.05, P<0.01). CCK-8 assay showed that the cells viability was significantly inhibited with drug intervention at the concentrations of 400 and 800 mg·L-1 (P<0.01). Western blot demonstrated that compared with the model group, ethyl acetate extract, n-butanol extract and chloroform extract decreased the expression levels of Col-Ⅰ, Col-Ⅲ, TIMP2 and CTGF in HK-2 cells induced by TGF-β1, and increased the expression of MMP-2 (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of immunofluorescence showed that ethyl acetate extract, n-butanol extract and chloroform extract could inhibit the expression of α-SMA (P<0.05), with more significant effect in n-butanol extract (P<0.01). The results of Real-time PCR showed that ethyl acetate extract and chloroform extract inhibited mRNA expression of PAI-1 (P<0.05), with more significant effect in n-butanol extract (P<0.01). Conclusion:The extracts of ethyl acetate, n-butanol and chloroform are the active parts of Taohe Chengqitang with the anti-renal fibrosis effect, with n-butanol extract as the most active part. The mechanism on anti-renal fibrosis may be related to its regulation of extracellular matrix (ECM) synthesis and degradation.

Effect of Fushengong Prescription on Wnt/β-catenin Signaling Pathway in Kidney of Rats with Chronic Renal Failure / 中国实验方剂学杂志

Objective:To observe the effects of Fushengong prescription on secretory glycoprotein （Wnt）/β-serial protein （β-catenin） signaling pathway in kidney of rats with chronic renal failure （CRF），and to further explore its mechanism of releasing the aggregation of extracellular matrix（ECM），inhibiting renal tubule interstitial fibrosis （TIF） and prolonging the progression of CRF. Method:A total of 55 SD male rats were randomly divided into the normal group，the model group，and the low， medium and high dose groups of Fushengong prescription，with 11 rats in each group.The normal group was routinely reared and the other 4 groups of rats were used to establish CRF model with 0.5% adenine fodder， fed them continuously for 21 d. After successful modeling，all model rats were switched to conventional feed. Normal saline （NS） was given the normal group and the model group by 20 mL·kg-1·d-1， the low， middle and high dose groups rats of Fushengong prescription were given intragastric administration Fushengong prescription according to the body weight of 4， 8， 16 g·kg-1，once a day，continuous gavage for 30 d. After the experiment，the pathomorphism change of renal tissues of rats was measured by Masson staining， the expression of Wnt4 and β-catenin mRNA in the kidney tissues were observed by the method of Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， the expression of Wnt4，β-catenin and matrix metalloproteinase-7（MMP-7） protein of renal tissues were detected by the methods of Western blot. The expression of Wnt4， β-catenin protein of renal tissues were detected by the methods of immunohistochemistry （IHC）. Result:Compared with normal group，renal tubule interstitial fibrosis of renal tissues increased distinctly and the expression of Wnt4，β-catenin and MMP-7 protein increased significantly in the model group. Wnt4 and β-catenin mRNA also increased significantly in model group（P<0.01）. Compared with model group， the expression of Wnt4， β-catenin and MMP-7 protein in the Fushengong prescription groups decreased obviously （P<0.05）. The expression of Wnt4 and β-catenin mRNA in Fushengong prescription groups also decreased obviously. Conclusion:The mechanism of Fushengong prescription can release the aggregation of ECM，inhibit TIF and delay the progression of CRF，which may be related with the activation of Wnt/β-catenin signal pathway.

Effect of mebt/mebo on expression of mmp-2 and mmp-9 in chronic refractory wounds / 中国药理学通报

Aim To investigate the mechanism of MEBT/MEBO in promoting chronic wound healing by MMP-2 and MMP-9. Methods Ninety Wistar rats were randomly divided into five groups: MEBO group, rb-bFGF group, chronic wound group, acute wound group and blank group. Eighteen rats in each group were modeled as chronic refractory wounds. The ex-pression of MMP-2 and MMP-9 in wound tissues, on 3rd and 14th day, was detected by ELISA, Western blot and qRT-PCR. Results ( 1 ) Compared with chronic wound group, the time of wound healing was obviously shorter and the rate of wound healing was higher in MEBO group and rb-bFGF group ( P < 0. 05) ; meanwhile, the growth of wounds and the pathological changes of wounds were better in MEBO group and rb-bFGF group. (2) On 3rd day, the ex pression of MMP-2 and MMP-9 in MEBO group and rb-bFGF group was higher than that in chronic wound group (P <0. 05) ; however, on 14th day, the expression of MMP-2 and MMP-9 in MEBO group and rb-bF-GF group also decreased (P < 0. 05). (3) Compared with chronic wound group, the expression of MMP-2 and MMP-9 in MEBO group and rb-bFGF group increased on 3rd day (P <0. 05) , while the expression of MMP-2 and MMP-9 decreased in MEBO group and rb-bFGF group on 14th day (P <0.05). Conclusions MEBT/MEBO promoting chronic wound healing may be associated with regulating MMP-2 and MMP-9 to participate in the degradation and remodeling of ECM.

Berberine prevents primary peritoneal adhesion and adhesion reformation by directly inhibiting TIMP-1

Peritoneal adhesions are fibrous tissues that tether organs to one another or to the peritoneal wall and represent the major cause of postsurgical morbidity. Enterolysis at repeat surgeries induces adhesion reformation that is more difficult to prevent than primary adhesion. Here we studied the preventive effects of different approaches of berberine treatment for primary adhesion, and its effects on adhesion reformation compared to Interceed. We found the primary adhesion was remarkably prevented by berberine through intraperitoneal injection 30 min before abrasive surgery (pre-berberine) or direct addition into injured cecum immediately after the surgery (inter-berberine). Rats with adhesion reformation had a more deteriorative collagen accumulation and tissue injury in abrasive sites than rats with primary adhesion. The dysregulated TIMP-1/MMP balance was observed in patients after surgery, as well as adhesion tissues from primary adhesion or adhesion reformation rats. Inter-berberine treatment had a better effect for adhesion reformation prevention than Interceed. Berberine promoted the activation of MMP-3 and MMP-8 by directly blocking TIMP-1 activation core, which was reversed by TIMP-1 overexpression in fibroblasts. In conclusion, this study suggests berberine as a reasonable approach for preventing primary adhesion formation and adhesion reformation.

Effect of Kangxianling Decoction on Renal Function and Kidney ECM Expression in Rats with Renal Fibrosis / 中国实验方剂学杂志

Objective: To investigate the effect of Kangxianling decoction on renal function and expression of extracellular matrix(ECM) in renal tissue of rats with renal fibrosis induced by 5/6 nephrectomy. Method: Totally 50 SD rats were randomly divided into control group (n=7), sham-operation group (n=7), operation group (n=36). The 5/6 nephrectomized renal fibrosis model was established in operation group. After successful modeling, rats were randomly divided into model group, Kangxianling group and losartan potassium group. The losartan potassium group and the Kangxianling group were given losartan potassium (33.3 g·kg-1) and Kangxianling decoction (21 g·kg-1) every day. The control group, sham-operation group and model group were all treated with equal volume of normal saline. Rats were put to death after 24 weeks of consecutive medication, and renal function, 24 h urine volume and 24 h-Pro quantitation of each group were measured. The expressions of collagen Ⅰ(ColⅠ) and ColⅢ in renal tissues were determined by immunohistochemistry and Western blot. Result: Compared with the normal group and sham-operation group, serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine volume, 24 h-Pro quantitation, renal tissue ColⅠ and ColⅢ expression levels were significantly increased in the model group (PPPConclusion: Kangxianling decoction can down-regulate the expressions of ColⅠ and ColⅢ in kidney tissue, there by inhibiting the accumulation of ECM, and exerting the effect in delaying renal fibrosis and protecting renal function.

PEP06 polypeptide 30 is a novel cluster-dissociating agent inhibiting v integrin/FAK/Src signaling in oral squamous cell carcinoma cells

Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies. In oral squamous cell carcinoma (OSCC), v integrin is a crucial mediator of multicellular clustering and collective movement ; however, its contribution to metastatic spread remains to be addressed. According to the emerging therapeutic concept, dissociation of tumor clusters into single cells could significantly suppress metastasis-seeding ability of carcinomas. This study aimed to investigate the anti-OSCC potential of novel endostatin-derived polypeptide PEP06 as a cluster-dissociating therapeutic agent . Firstly, we found marked enrichment of v integrin in collectively invading multicellular clusters in human OSCCs. Our study revealed that metastatic progression of OSCC was associated with augmented immunostaining of v integrin in cancerous lesions. Following PEP06 treatment, cell clustering on fibronectin, migration, multicellular aggregation, anchorage-independent survival and colony formation of OSCC were significantly inhibited. Moreover, PEP06 suppressed v integrin/FAK/Src signaling in OSCC cells. PEP06-induced loss of active Src and E-cadherin from cell-cell contacts contributed to diminished collective migration of OSCC . Overall, these results suggest that PEP06 polypeptide 30 inhibiting v integrin/FAK/Src signaling and disrupting E-cadherin-based intercellular junctions possesses anti-metastatic potential in OSCC by acting as a cluster-dissociating therapeutic agent.

Effects of Matrix Stiffness on Proliferation and Glucose Metabolism of Hepatocellular Carcinoma Cells / 医用生物力学

Objective To investigate the influences of different matrix stiffness on proliferation ability and glucose metabolism of hepatocellular carcinoma (HCC) cells and to explore the correlation between metabolism and biological behavior changes of HCC cells resulted from the stiffness of extracellular matrix (ECM)．Methods The proliferation changes of HepG2 cells cultured on matrix with different stiffness were detected by CCK-8 assay and cell count assay. 2-NBDG and flow cytometry were used to detect the effect of matrix stiffness on glucose uptake. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of Glut1. Then, 2-DG was used to inhibit glycolysis, and the influences of matrix stiffness on proliferation of HepG2 cells were detected. Results The proliferation ability, glucose uptake and the expression of Glut1 of HepG2 cells increased with the matrix stiffness increasing. When glycolysis was inhibited, the proliferation ability of HepG2 cells grown on matrix with different stiffness was similar. Conclusions The mechanical microenvironment had an important effect on proliferation of HCC cells; matrix with a larger stiffness might promote proliferation of HCC cells through regulating glycolysis. The research findings provide a corresponding experimental basis for the clinical treatment of HCC cells and drug development targeting glucose metabolism.