Knockdown of the long non­coding RNA MALAT1 ameliorates TNF­α­mediated endothelial cell pyroptosis via the miR­30c­5p/Cx43 axis.

Long noncoding RNAs (lncRNAs) are related to the development of atherosclerosis (AS). However, the role of lncRNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) in tumor necrosis factor­α (TNF­α)­induced rat aortic endothelial cell (RAOEC) pyroptosis, as well as the underlying mechanisms, remain unclear. RAOEC morphology was assessed using an inverted microscope. The mRNA and/or protein expression levels of MALAT1, microRNA(miR)­30c­5p and connexin 43 (Cx43) were assessed using reverse transcription­quantitative PCR (RT­qPCR) and/or western blotting, respectively. The relationships among these molecules were validated by dual­luciferase reporter assays. Biological functions, such as LDH release, pyroptosis­associated protein levels and the proportion of PI­positive cells, were evaluated using a LDH assay kit, western blotting and Hoechst 33342/PI staining, respectively. The present study demonstrated that compared with the control group, the mRNA expression levels of MALAT1 and protein expression levels of Cx43 were significantly up­regulated, whereas miR­30c­5p mRNA expressions levels were significantly decreased in TNF­α­treated RAOEC pyroptosis. Knockdown of MALAT1 or Cx43 significantly attenuated the increase in LDH release, pyroptosis­associated protein expression and PI­positive cell numbers among RAOEC treated using TNF­α, whereas an miR­30c­5p mimic exerted the opposite effect. Furthermore, miR­30c­5p was demonstrated to be a negative regulator of MALAT1 and could also target Cx43. Finally, co­transfection with siMALAT1 and miR­30c­5p inhibitor could attenuate the protective effect of MALAT1 knockdown against TNF­α­mediated RAOEC pyroptosis by upregulation of Cx43 expression. In conclusion, MALAT1 might serve an important role in TNF­α­mediated RAOEC pyroptosis by regulating the miR­30c­5p/Cx43 axis, which would provide a potential novel diagnostic and therapeutic target for AS.

Exosomes Derived from Glioma Cells under Hypoxia Promote Angiogenesis through Up-regulated Exosomal Connexin 43.

Glioblastoma multiform (GBM) is a highly aggressive primary brain tumor. Exosomes derived from glioma cells under a hypoxic microenvironment play an important role in tumor biology including metastasis, angiogenesis and chemoresistance. However, the underlying mechanisms remain to be elucidated. In this study, we aimed to explore the role of connexin 43 on exosomal uptake and angiogenesis in glioma under hypoxia. U251 cells were exposed to 3% oxygen to achieve hypoxia, and the expression levels of HIF-1α and Cx43, involved in the colony formation and proliferation of cells were assessed. Exosomes were isolated by differential velocity centrifugation from U251 cells under normoxia and hypoxia (Nor-Exos and Hypo-Exos), respectively. Immunofluorescence staining, along with assays for CCK-8, tube formation and wound healing along with a transwell assay were conducted to profile exosomal uptake, proliferation, tube formation, migration and invasion of HUVECs, respectively. Our results revealed that Hypoxia significantly up-regulated the expression of HIF-1α in U251 cells as well as promoting proliferation and colony number. Hypoxia also increased the level of Cx43 in U251 cells and in the exosomes secreted. The uptake of Dio-stained Hypo-Exos by HUVECs was greater than that of Nor-Exos, and inhibition of Cx43 by 37,43gap27 or lenti-Cx43-shRNA efficiently prevented the uptake of Hypo-Exos by recipient endothelial cells. In addition, the proliferation and total loops of HUVECs were remarkably increased at 24 h, 48 h, and 10 h after Hypo-Exos, respectively. Notably, 37,43gap27, a specific Cx-mimetic peptide blocker of Cx37 and Cx43, efficiently alleviated Hypo-Exos-induced proliferation and tube formation by HUVECs. Finally, 37,43gap27 also significantly attenuated Hypo-Exos-induced migration and invasion of HUVECs. These findings demonstrate that exosomal Cx43 contributes to glioma angiogenesis mediated by Hypo-Exos, and suggests that exosomal Cx43 might serve as a potential therapeutic target for glioblastoma.

Study on the relationship between vascular endothelial growth factor and syndrome type of traditional Chinese medicine in patients with gastric carcinoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>By detecting the expression of vascular endothelial growth factor (VEGF) in different syndrome types of traditional Chinese medicine (TCM) in patients with gastric carcinoma (GC) to investigate the correlation of VEGF and TCM syndrome type for exploring the internal relationship between disease and syndrome.</p><p><b>METHODS</b>The correlated clinical manifestations were recorded in detail and patient's syndrome type was differentiated at beside before operation. After operation, pathological examination on the resected specimen was performed routinely. The expression of VEGF in 104 GC patients of three different syndrome types, i.e. Pi-Wei asthenia type (type 1), Gan-Wei disharmony type (type 2) and phlegm-stasis-poisons stagnant type (type 3)and in 30 patients with superficial gastritis (for control) were determined by immunohistochemical method. The expressions of VEGF in different syndrome types were analyzed by combining with correlated clinical pathological characteristics.</p><p><b>RESULTS</b>(1) The VEGF protein positive expression rate in GC patients was 81.7% (85/104), it was significantly higher than that in the control group with positive rate being zero (P < 0.01). (2) The intensity of VEGF expression was positively correlated with the depth of tumor infiltration, the extent of lymph node metastasis and the pathological staging of cancer (P < 0.01), and showed no evident correlation with the size of tumor and its degree of cell differentiation (P > 0.05). (3) Levels of VEGF expression in various syndrome types were significantly different (P < 0.01), the highest expression showed in the type 3, the second in the type 2, and the lowest in the type 1, comparison between them all showed significant difference (P < 0.01 for 3 vs 1, P < 0.05 for 3 vs 2 and 2 vs 1).</p><p><b>CONCLUSION</b>(1) VEGF could be taken as an index for evaluating the infiltration, metastasis and prognosis of GC. (2) The different expressions of VEGF play different roles in the GC metastatic process of patients of different syndrome types, and the pathogenesis of GC metastasis may be probably various in different syndrome types. VEGF is the relevant gene of GC of type 2 and type 3. (3) High VEGF expression shows positive correlation with the GC metastasis of type 2 and 3, suggesting that the higher the intensity of evil excess, the more possibilities of tumor infiltration and metastasis and the worse the prognoses will be.</p>

Relationship between sCD44v6 expression and TCM differentiation type of gastric carcinoma patients and influence of weitai capsule on the expression / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the relationship of TCM type with serum level of soluble CD44v6 (sCD44v6) and different histologic parameters in gastric carcinoma patients and to observe the influence of Weitai capsule (WTC) on the sCD44v6 expression.</p><p><b>METHODS</b>TCM typing and sCD44v6 expression were determined in all the enrolled patients (30 in the control and 32 in the trial group) before operation, and 3-4 courses of chemotherapy was applied to them from 3-4 weeks after operation. To the patients of trial group, oral administration of WTC was given additionally with 4 capsules, 3 times a day for consecutive 3 months.</p><p><b>RESULTS</b>sCD44v6 was significantly positive correlated with the degree of cancer cell differentiation, infiltration and lymph node metastasis; (2) Level of sCD44v6 was the highest in patients of blood stasis type, as compared with that in the patients of Pi-deficiency type or of damp-heat type, the difference was significant; (3) After ending treatment, level of sCD44v6 in the trial group was significantly lower than that in the control group.</p><p><b>CONCLUSION</b>(1) Serum level of sCD44v6 could be taken as the criterion for evaluating the development and prognosis of gastric cancer, as well as the therapeutic target for anti-metastasis treatment; (2) Serum level of sCD44v6 is related to some extent with TCM type of blood stasis and Pi-deficiency; (3) WTC combined with chemotherapy could further inhibit the expression of serum sCD44v6 in gastric carcinoma patients.</p>

Clinical study on effect of yuantong capsule in treating vascular dementia / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the clinical effect of Yuantong Capsule (YTC) in treating vascular dementia (VD).</p><p><b>METHODS</b>Eighty-three patients of VD were randomized on ratio of 2:1 into two groups, the 54 patients in the treated group were treated with YTC orally administered, 3 times a day, 1 capsule in each time. The remaining 29 patients in the control group were treated with Hydergine orally, 3 times a day, 2 mg in each time. The therapeutic course for both groups was 2 months.</p><p><b>RESULTS</b>The therapeutic effect in the treated group was significantly better than that in the control group, significant difference (P < 0.05 or P < 0.01) was shown in comparison of the two groups in terms of the mini-mental state examination (MMSE) and activity of daily living (ADL) test, symptoms scoring, total effective rate, and laboratory indexes findings.</p><p><b>CONCLUSION</b>The therapeutic effect of YTC in treating VD was obvious.</p>