AIM　Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with β-adrenoceptor(β-AR) and G protein subunit(Gsα),and the interfering action of anisodamine. METHODS　PMVEC was isolated and cultured from Wistar rat in vitro, radioligand binding assay and flow cytometer were used. RESULTS　After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously ,while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of β-AR and Gsα protein level. Anisodamine can inhibit these changes above. CONCLUSION　LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolayer by influencing two links of both β-AR and Gsα protein.

In order to probe the effects of lipopolysaccharide(LPS) on ? adrenoceptor (? AR) and ? AR kinases in rat pulmonary microvascular endothelial cells (RPMVEC), radioactive ligand binding assay was used to measure the distribution density of ? AR before and after LPS acting in it; Western blot was used to observe the expression changes in ? AR kinases, including GRK2 and GRK3. The results showed that the maximum binding capacity (Bmax) of RPMVEC ? AR was 5 583?0 306 fmol/10 5 cells. The Bmax of ? AR in LPS group significantly decreased compared with normal control group ( P

AIM Observing the influence of lipopolysaccharide (LPS) on changes of the permeability and actin cytoskeleton of rat pulmonary microvascular endothelial cells (PMVEC) monolayer, we explore their relationship with ?-adrenoceptor(?ＡＲ) and G protein subunit(Gs?), and the interfering action of anisodamine. METHODS PMVEC was isolated and cultured from Wistar rat in vitro, radi- oligand binding assay and flow cytometer were used. RESULTS After incubation of LPS in vitro, the central F-actin of PMVEC depolymerized and its density decreased obviously , while the permeability of PMVEC monolyer increased significantly. Meanwhile it was found that LPS can induce the down regulation of 5-AR and Gsa protein level. Anisodamine can in- hibit these changes above. CONCLUSION LPS can directly cause the increase of permeability of PMVEC monolayer which relates to the depolymerization of central F-actin in PMVEC. Anisodamine may take part in regulating actin cytoskeleton of PMVEC and attenuate LPS-induced the increase of permeability of PMVEC monolaver by influencing two links of both ?-AR and Gsa protein

Objective To investigate the effects of tumor necrosis factor ? (TNF ?) on ? adrenoceptor (? AR) and ? AR related G protein coupled receptor kinases (GRKs) in rat pulmonary microvascular endothelial cells (PMVECs) as well as the interfering action of anisodamine. Methods Radio ligand binding assay was used to measure the maximal binding capacity (B max ) changes of ? AR in rat PMVECs after treatment with TNF ?. The effects of TNF ? and ? AR related GRKs expression at the protein level were observed by Western blotting. Results B max of ? AR in normal rat PMVECs was (5.58?0.31) fmol/10 5 cells. B max of ? AR in TNF ? group decreased significantly as compared with that in the normal control group, but no significant difference was found between the normal control group and TNF ?+anisodamine group. The expression of GRK2 in rat PMVECs was positive, but expression of GRK3, GRK5, and GRK6 were negative. The expression of GRK2 in TNF ? group and TNF ?+anisodamine group increased significantly as compared with that in the normal control group, but no significant difference was found between the TNF ? group and the TNF ?+anisodamine group. Conclusion GRK2 but not GRK3, GRK5, or GRK6 is expressed in rat PMVECs. The increased expression of GRK2 induced by TNF ? in rat PMVECs might promote the phosphorylation of ? AR, leading to ? AR internalization and decoupling with G protein, which might be the main mechanism of down regulation of ? AR induced by TNF ?. Anisodamine might inhibit the down regulation of ? AR through other mechanism instead of inhibiting the increase of GRK2 expression.

ObjectiveTo evaluate the feasibility of constructing tissue engineering cardiac patch with photooxidationfixed acellular bovine pericardium.MethodsFresh bovine pericardia were treated by dye-mediated photooxidation after decellularization.Some of them were seeded with bone marrow stromal cells(MSCs) isolated from male SD rats to construct cardiac patches.Myocardial infarction(MI) model was made in female SD rats by left anterior descending coronary ligation(LAD).One week later, the confirmed MI rats were divided into three groups randomly, group MI (n = 15)without any treatment; group P (n = 18) with photooxidated pericardia implantation ; group P + C (n = 18) with seeded pericardia implantation.A sham group (n = 10) was also performed with opening and closing chest twice only.The heart were explanted at 2 or 4 weeks after implantation, and examined histologically and immunohistochemically.The heart function was evaluated by echocardiography at 4 weeks before excising the rats.ResultsThere were no cells or cell debris remained in bovine pericardium tissue.The fiber structure became condensed after photooxidation.The seeded cells formed a continuous layer on the surface of the tissue.The pericardial degradation level and newly formed microvessel density were larger in group P + C than in group P after 2 [ (13.7 ±5.2)个/mm2 vs (7.1 ±3.1)个/mm2, P＜0.05]and4 [(22.6 ±4.9)个/mrn2 vs (14.1 ±5.3)个/mm2, P＜0.05]weeks.Four weeks after transplantation, cardiac echocardiography showed left ventricular ejection fraction(LVEF) was lower in group MI (44.8 ± 4.4) ％ and group P (48.4 ± 5.0) ％ compared with group P + C (49.3 ± 4.8) ％, left ventricular fractional shorterning(LVFS) was lower in group MI (18.0 ± 2.2) ％ and group P (19.8 ± 2.5) ％ compared with group P + C (20.4 ±2.5) ％, the difference between P + C and MI was significant.ConclusionTransplantation of the tissue engineered bovine pericardial patches with dye-mediated photooxidation can improve heart function in MI rats.This kind of patches demonstrates a promising prospect in the future.

Pyrrole [1,2-α] indole is a novel fused heterocyclic skeleton, which is also the basic structural unit and synthetic intermediate of many natural active products and drugs. Pyrrole [1,2-α] indole heterocyclic derivatives have attracted much attention in organic synthesis and medicinal chemistry because of their extensive and marked biological activities. Plant extracts have always been an important source of active compounds. At present, the alkaloids based on the pyrrole [1,2-α] indole heterocyclic structure discovered and isolated from plant extracts include isatisine, isoborreverine, flinderoles, polyavolensin and yuremamine. This paper reviews the research progress on the biological activity of pyrrole [1,2-α] indole heterocyclic derivatives and has found that pyrrole [1,2-α] indole heterocyclic derivatives have a good development prospect in screening active compounds and developing candidate drugs.

OBJECTIVETo screen novel miRNAs targeting EZH2 3' untranslated region (UTR) in recombinational MCF-7 breast cancer cells over-expressing EZH2 3' UTR and quantitative analyze the expressions of the screened miRNA in breast cancer cells and tissues.

METHODSA lentiviral library was transfected into the recombinant cell line MCF-7. The cells were screened with cytotoxic agents before extraction of the genome for amplification of the miRNA precursors using PCR. The screened miRNAs were identified with sequence analysis and their expressions were analyzed quantitatively with real-time PCR in breast cancer cells and tissues.

RESULTSSeven miRNAs were screened from the recombinant MCF-7 cells, namely miR-15b, miR-16-2, miR-181b2, miR-217, miR-224, miR-329-1, and miR-487b, all of which failed to be predicted by bioinformatics software. Real-time PCR showed that miR-217, miR-329-1, and miR-487b were over-expressed in MCF-7 cells, and the expression of miR-15b and miR-16-2 was significantly increased in cancer tissues compared with the adjacent tissues (P<0.05).

CONCLUSIONNovel targeted miRNAs that can not be predicted by bioinformatics software were successfully screened from MCF-7 breast cancer cells over-expressing EZH2 3' UTR. These miRNAs are expressed differentially between normal breast cells and breast cancer tissues.

3' Untranslated Regions ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Female ; Gene Expression Profiling ; Humans ; Lentivirus ; genetics ; MicroRNAs ; genetics ; Polycomb Repressive Complex 2 ; genetics

Gliomas are a group of refractory heterogeneous diseases. Gliomas even at the same pathological type and grade exhibit different outcomes after treatment. Therefore, glioma patients have obvious survival difference. The development of intraoperative auxiliary means has greatly contributed to surgical resection of glioma and remarkably increased therapeutic effects. The development of sensitizer realizes the combination of radiotherapy and chemotherapy and can improve the anti-tumor effects of oral temozolomide. Molecular markers and signal pathways involved in glioma, such as isocitrate dehydrogenase-1 mutation, epidermal growth factor amplification, high expression of vascular endothelial growth factor, Notch signal pathway, miRNA, etc. are involved in the occurrence and development of glioma and have an obvious impact on the proliferation, metastasis and invasion of glioma. They are potential molecular targets for the clinical treatment of glioma. Many different immunotherapy schemes are actively carried out in patients with glioma, but the unique tumor immune microenvironment of the central nervous system needs to be considered. This paper reviews the treatment progress of glioma in recent years.

Acute myeloid leukemia is a clonal malignant proliferative disease of myeloid blasts of the hematopoietic system. The leukemia cell population is composed of cells of different stages. Acute myeloid leukemia stem cells are the cells that may cause diseases in immunodeficient animals and can regenerate themselves through continuous transplantation, which causes leukemia. Since more than 96% of leukemia stem cells are in the G0 stage, they may escape the effects of chemical drug stabbing, leading to drug resistance and recurrence of leukemia. Therefore, the key to the treatment of acute myeloid leukemia has always been how to remove leukemia stem cells without damaging hematopoietic stem cells. This review paper addresses the new developments in the immunophenotype of leukemia stem cells and leukemia stem cells-targeting therapy for acute myeloid leukemia.

MEK is a canonical effector of mutant KRAS; however, MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers. Here, we identified mitochondrial oxidative phosphorylation (OXPHOS) induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer (NSCLC) resistance to the clinical MEK inhibitor trametinib. Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment, satisfying their energy demand and protecting them from apoptosis. As molecular events in this process, the pyruvate dehydrogenase complex (PDHc) and carnitine palmitoyl transferase IA (CPTIA), two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation. Importantly, the co-administration of trametinib and IACS-010759, a clinical mitochondrial complex I inhibitor that blocks OXPHOS, significantly impeded tumor growth and prolonged mouse survival. Overall, our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.