Geniposide promotes wound healing of skin ulcers in diabetic rats through PI3K/Akt pathway.

Continuously hyperglycation-induced lesion and poor blood flow contributed to the wound incurable and susceptible to infection. About fifteen percent of people with diabetes would develop ulcers during their lifetime, especially on the feet, which could lead to severe tissue destruction and eventual amputation. Various strategies were limited to accelerate wound healing in diabetic patients for high cost and unsatisfied effects. Geniposide is well-known for its anti-inflammation and anti-apoptosis in several pathological tissues. This study is to explore the protective effect of geniposide on wound healing rate, inflammatory response, nutritional function and cellular apoptosis in diabetic rats. Diabetic rats was induced by streptozotocin and defined as plasma glucose >300 mg/dl. Western blot and immunostaining technologies were performed to mark and quantify the target proteins. The oral administration of geniposide (200 mg/kg and 500 mg/kg) could significantly promote wound healing by the increment of lesion retraction in diabetic rats compared to model group. In the apoptotic study of skin wound in diabetic rats, the TUNEL-positive cells were greatly decreased in geniposide subgroups (P < 0.05). The levels of TNF-α, IL-1ß and IL-6 were significantly inhibited by geniposide with the IC50 value of 470 mg/kg, 464 mg/kg and 370 mg/kg body weight respectively, which might be related to the enhancement of the phosphorylation of PI3K and Akt proteins. Geniposide enhanced the repairment of skin wound in diabetic rats by inhibiting inflammatory response and apoptosis.

Antithrombotic effects of Huanglian Jiedu decoction in a rat model of ischaemia-reperfusion-induced cerebral stroke.

CONTEXT: Huanglian Jiedu Decoction (HLJJD) has a variety of pharmacological activities, such as anti-inflammatory and neuroprotection against ischaemic brain injury. OBJECTIVES: This ex vivo study explores its antithrombosis activity and inhibition of platelet aggregation. MATERIAL AND METHODS: To study the antithrombosis activity of HLJJD ex vivo, saline, or HLJDD (100, 200, and 500 mg/kg) was treated prophylactically by gavage for 3 days in Wistar rats (n = 4). Based on the rat model of transient middle cerebral artery infarction (MCAO) or normal rats (n = 4), the antithrombotic activity in the normal group and HLJDD subgroups on prothrombin time, thrombus weight, platelet aggregation, and others was evaluated, followed by the antiplatelet aggregation of its main components (n = 4). RESULTS: The weight of the thrombus increased significantly at 24 h after MCAO onset. HLJJD did not influence the change of PT, but significantly inhibited thrombosis by 12.5, 20.0, and 20.5% in reducing the dry weight of thrombus, and blocked collagen-induced platelet aggregation by 25.5, 39.0, and 42.7% and adhesion of blood platelet by 17.3, 26.2, and 27.3%. The IC50 value of HLJJD on collagen-induced platelet aggregation was 670 mg/kg. Geniposide only facilitated antiplatelet aggregation induced by collagen, but not AA or ADP. Both baicalin and berberine showed markedly antiplatelet aggregation induced by all activators. The antithrombotic activity of baicalin was relatively higher than that of berberine (35.0-47.8% vs. 20.6-33.5%). CONCLUSION: Our results indicated that HLJDD regulated blood circulation by inhibiting platelet aggregation and thrombosis, which might also extensively contribute to the clinical prevention and treatment of cerebrovascular diseases.

A novel multifunctional gold nanorod-mediated and tumor-targeted gene silencing of GPC-3 synergizes photothermal therapy for liver cancer.

Tumor-specific targeted delivery is a major obstacle to clinical treatment of hepatocellular carcinoma (HCC). Here we have developed a novel multi-functional nanostructure GAL-GNR-siGPC-3, which consists of Galactose (GAL) as the HCC-targeting moiety, golden nanorods (GNR) as a framework to destroy tumor cells under laser irradiation, and siRNA of Glypican-3 (siGPC-3) which induce specifically gene silence of GPC-3 in HCC. Glypican-3 (GPC-3) gene is highly associated with HCC and is a new potential target for HCC therapy. On the other hand, Gal can specifically bind to the asialoglycoprotein receptor which is highly expressed on membrane of hepatoma cells. GAL and siGPC-3 can induce targeted silencing of GPC-3 gene in hepatoma cells. In vivo and in vitro results showed that GAL-GNR-siGPC-3 could significantly induce downregulation of GPC-3 gene and inhibit the progression of HCC. More notably, GAL-GNR-siGPC-3 could induce both GPC-3 gene silencing and photothermal effects, and the synergistic treatment of tumors was more effective than individual treatments. In summary, GAL-GNR-siGPC-3 achieved a synergistic outcome to the treatment of cancer, which opens up a new approach for the development of clinical therapies for HCC.

Isolation, Purification and Antioxidant Activity of The Polysaccharides from Chinese Truffle Tuber sinense.

The content of polysaccharides in Tuber sinense was investigated by isolation and purification, followed with the further antioxidant studies in total reducing capacity and radical scavenging activities. The crude extract of polysaccharides was purified by dialysis, column chromatography, and High Performance Liquid Chromatography. The main components of monosaccharide (s) and molecular structure of single polysaccharide were studied by using methylation, GC-MS, and NMR analysis. One new water-soluble non-starch polysaccharide (PTS-A with the yield of 0.41%) from T. sinense was purified and identified on structural characteristics for the first time. The characterizations of PTS-A were studied on physicochemical properties, main components of monosaccharide (s) and molecular structure. PTS-A was identified as glucan, only containing D-glucoses with the molecular structure of [→6) α-D-Glcp (1→6) α-D-Glcp (1→]n by methylation analysis and NMR. In the determination of total reducing capacity, their reducing abilities could be listed as vitamin C> PTS-A> crude polysaccharides-3> crude polysaccharides-2> crude polysaccharides-1. All of PTS-A, crude polysaccharides-2 and -3 were relatively good scavenger for 1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1- (2,4,6-trinitrophenyl) hydrazyl radicals with the IC50 of 2.81, 4.17 and 3.44 mg/mL, respectively. Thus, the separation and purification of polysaccharides were significant to increase the antioxidant activity in some degree. One new water-soluble 1,6-α-ᴅ-dextran was discovered with the polysaccharide structure identified for the first time. Both PTS-A and crude extracts of polysaccharide performed a potent potential on antioxidant activities. The bioactivities of PTS-A should be generalized to the broader pharmacological effects.

Silencing IDO2 in dendritic cells: A novel strategy to strengthen cancer immunotherapy in a murine lung cancer model.

While dendritic cell (DC)­based immunotherapy has achieved satisfactory results in animal models, its effects were not satisfactory as initially expected in clinical applications, despite the safety and varying degrees of effectiveness in various types of cancer. Improving the efficacy of the DC­based vaccine is essential for cancer immunotherapy. The present study aimed to investigate methods with which to amplify and enhance the antitumor immune response of a DC­based tumor vaccine by silencing the expression of indoleamine 2,3­dioxygenase 2 (IDO2), a tryptophan rate­limiting metabolic enzyme in DCs. In vitro experiments revealed that the silencing of IDO2 in DCs did not affect the differentiation of DCs, whereas it increased their expression of costimulatory molecules following stimulation with tumor necrosis factor (TNF)­α and tumor lysate from Lewis lung cancer (LLC) cells. In a mixed co­culture system, the IDO2­silenced DCs promoted the proliferation of T­cells and reduced the induction of regulatory T­cells (Tregs). Further in vivo experiments revealed that the silencing of IDO2 in DCs markedly suppressed the growth of tumor cells. Moreover, treatment with the IDO2­silenced DC­based cancer vaccine enhanced cytotoxic T lymphocyte activity, whereas it decreased T­cell apoptosis and the percentage of CD4+CD25+Foxp3+ Tregs. On the whole, the present study provides evidence that the silencing of the tryptophan rate­limiting metabolic enzyme, IDO2, has the potential to enhance the efficacy of DC­based cancer immunotherapy.

Effect of salidroside on bone marrow haematopoiesis in a mouse model of myelosuppressed anaemia.

This study was designed to investigate the effect of salidroside (SAL) on bone marrow haematopoiesis in a mouse model of myelosuppressed anemia. After the mouse model was established by 60Co γ irradiation and cyclophosphamide, pancytopenia and a sharp reduction in bone marrow stromal cells and bone marrow haematopoietic stem cells (lineage-Sca1+c-kit+) were observed. This was greatly alleviated by SAL (25 mg/kg, 50 mg/kg, 100 mg/kg) in a dose-dependent manner (50% effective dose value of 35.7 mg/kg and 61.2 mg/kg, respectively), followed by a distinct increment in anti-apoptotic protein Bcl-2. For cell culture in vitro, treatment with SAL resulted in a significant recovery of burst-forming unit-erythroids, and colony-forming unit-granulocyte macrophages on Day 7, and colony-forming unit-erythroids on Day 3, dose-dependently, but not of colony-forming unit-megakaryocyte macrophages. Inoculation of bone marrow cells derived from SAL-administrated donor mice resulted in a 60% survival of recipient mice at the high dose of 100 mg/kg SAL at 2 months after surgery. SAL appeared to be able to stimulate the restoration of bone marrow haemopoietic regulation in myelosuppressed anemia. Based on the downregulation of Fas ligand associated with the expression of Caspase-3 at the protein level, it was suggested that SAL might have an anti-apoptotic effect on bone marrow cells in the Fas-apoptotic pathway of Fas/FasL-caspase-3.

Potential biocontrol efficacy of Trichoderma atroviride with cellulase expression regulator ace1 gene knock-out.

The biocontrol function of the repressor of cellulase expression I (ACE1) in Trichoderma atroviride was verified through constructing Δace1 mutant strain by Agrobacterium tumefaciens-mediated transformation. The activities of cell wall-degrading enzymes (cellulase, xylanase, chitinase, ß-1,3-glucanase, and protease) in the supernatant of Δace1 mutant strain were distinctly higher than those of control strain, followed with the elevation of related genes transcript levels. Besides, the Δace1 mutant resulted in an elevating transcript level of xyr1, but no obvious change in the expression of cre1, which suggested that ACE1 was negative regulator of the xyr1 transcription, but not involved in cre1 transcription. On core polyketide synthases of four biosynthesis gene clusters for antibiotic secondary metabolites, only the transcription levels of encoding genes Try83179/TryH and Aza79482/AzaJ in Δace1 mutant strain were significantly higher than that in wild-type during antagonizing with pathogenic fungi Fusarium oxysporum and Rhizoctonia solani (with the inhibition rate of 30.7 and 19.8%, respectively). The biocontrol function of Δace1 mutant strain was remarkably enhanced. The results indicated that ACE1, indeed, acted as a repressor for cell wall-degrading enzymes and PKSs expression in T. atroviride, and the Δace1 mutant strain effectively made related enzymes activities improved with potential enhancement of biocontrol potency.