LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway.

BACKGROUND: Non-small cell lung cancer (NSCLC) is the most common type of human lung cancers, which has diverse pathological features. Although many signaling pathways and therapeutic targets have been defined to play important roles in NSCLC, limiting efficacies have been achieved. METHODS: Bioinformatics methods were used to identify differential long non-coding RNA expression in NSCLC. Real-time RT-PCR experiments were used to examine the expression pattern of lncRNA PKMYT1AR, miR-485-5p. Both in vitro and in vivo functional assays were performed to investigate the functional role of PKMYT1AR/miR-485-5p/PKMYT1 axis on regulating cell proliferation, migration and tumor growth. Dual luciferase reporter assay, fluorescent in situ hybridization (FISH), immunoblot, co-immunoprecipitation experiments were used to verify the molecular mechanism. RESULT: Here, we identify a human-specific long non-coding RNA (lncRNA, ENST00000595422), termed PKMYT1AR (PKMYT1 associated lncRNA), that is induced in NSCLC by Yin Yang 1 (YY1) factor, especially in cancerous cell lines (H358, H1975, H1299, H1650, A549 and SPC-A1) compared to that in normal human bronchial epithelium cell line (BEAS-2B). We show that PKMYT1AR high expression correlates with worse clinical outcome, and knockdown of PKMYT1AR inhibits tumor cell proliferation, migration and xenograft tumor formation abilities. Bioinformatic analysis and a luciferase assay demonstrate that PKMYT1AR directly interacts with miR-485-5p to attenuate the inhibitory role on its downstream oncogenic factor PKMYT1 (the protein kinase, membrane-associated tyrosine/threonine 1) in NSCLC. Furthermore, we uncover that miR-485-5p is downregulated in both cancerous cell lines and peripheral blood serum isolated from NSCLC patients compared to reciprocal control groups. Consistently, forced expression of miR-485-5p inhibits the proliferation and migration abilities of tumor cells. Moreover, we provide evidence showing that PKMYT1AR targeting antisense oligonucleotide (ASO) dramatically inhibit tumor growth in vivo. Mechanistic study shows that PKMYT1AR/ miR-485-5p /PKMYT1 axis promotes cancer stem cells (CSCs) maintenance in NSCLC via inhibiting ß-TrCP1 mediated ubiquitin degradation of ß-catenin proteins, which in turn causes enhanced tumorigenesis. CONCLUSIONS: Our findings reveal the critical role of PKMYT1AR/miR-485-5p /PKMYT1 axis during NSCLC progression, which could be used as novel therapeutic targets in the future.

Long-Zhi Decoction Medicated Serum Promotes Angiogenesis in Human Umbilical Vein Endothelial Cells Based on Autophagy.

Ischemic stroke (IS) is a fatal subtype of stroke that lacks effective treatments. Angiogenesis following IS is an effective response that mediates brain recovery and repair. Our previous study demonstrated that long-zhi decoction (LZD), a Chinese herbal formula, promoted angiogenesis in rats of IS model. To further investigate the association between the proangiogenic mechanism of an LZD-medicated serum and cellular autophagy, we evaluated its promotional effect on angiogenesis in human umbilical vein endothelial cells (HUVECs) in vitro. We used HUVECs subjected to H2O2 to induce injury and observed the effects of the LZD-medicated serum treatment. Cell-based assays included proliferation, migration, and tube formation. To assess the extent of autophagy, transmission electron microscopy was used to measure the number of autophagosomes. Immunofluorescence and Western blotting were performed to evaluate the autophagy-related protein of LC3-II and Beclin-1. The LZD-medicated serum promoted proliferation, migration, and tube formation in HUVECs. The LZD-medicated serum also increased the autophagosomes and the autophagic protein expressions of LC3-II and Beclin-1. The proangiogenic and autophagic activity of LZD provides new cogitations to its clinical application and may lead to potential drug development for treating various vascular diseases, especially in the elderly, in the future.

Astragaloside IV attenuates Toll-like receptor 4 expression via NF-κB pathway under high glucose condition in mesenchymal stem cells.

Diabetic hyperglycemia causes a variety of pathological changes. Astragaloside IV (AS-IV) was widely used for the treatment of cardiovascular diseases in China. The aim of this study was to determine the effect of AS-IV on bone marrow mesenchymal stem cells (MSCs) and the underlying mechanism in diabetes. We used reverse transcription polymerase chain reaction and western blotting to determine the expression of Toll-like receptor 4 (TLR4), matrix metalloproteinase-2 (MMP-2) and NF-κB p65 in MSCs under high glucose (HG) with or without pretreatment with AS-IV. The surface expression of TLR4 was checked by flow cytometry and the expression of TNF-α and MCP-1 were detected by ELISA in diabetes patients treated with AS-IV. AS-IV promoted the proliferation of MSCs and attenuated the increased expression of TLR4 induced by HG. In addition, AS-IV decreased the HG-induced translocation of NF-κB p65 and increased the MMP-2 expression in MSCs. AS-IV decreased the TLR4, TNF-α and MCP-1 expression in patients. Collectively,our data revealed that AS-IV attenuated TLR4 expression through the NF-κB signaling pathway in MSCs.

[Observation on therapeutic effect of type II early diabetic nephropathies intervened by acupoint thread embedding].

OBJECTIVE: To observe the therapeutic effect of type II early diabetic nephropathies intervened by acupoint thread embedding for strengthening spleen and benefiting kidney. METHODS: Sixty cases of type II early diabetic nephropathies were randomly divided into an acupoint thread embedding group and a routine therapy group, 30 cases in each group. In routine therapy group, western medicine routine therapy was applied to control blood sugar, blood pressure and blood lipid. In acupoint thread embedding group, besides the western medicine routine therapy, thread embedding intervention was given at Pishu (BL 20), Zusanli (ST 36), Shenshu (BL 23) and Yishu (Extra) on both sides as main acupints, and the treatment course was 3 months. Urinary micro-albumin excretion (UAER), total score of TCM syndrome and monitor control indices (blood sugar, blood pressure, blood lipid, urea nitrogen and serum creatinine) in both groups were observed before and after treatment, and the therapeutic effects in both groups were compared. RESULTS: After treatment, the indices of UAER and total score of TCM syndrome were all reduced (P < 0.001, P < 0.01); the reduction in acupoint thread embedding group was more obvious (P < 0.01, P < 0.05); the total effective rate in acupoint thread embedding group was 76.7% (23/30), superior to that of 63.3% (19/30) in routine therapy group (P < 0.05). The blood sugar, blood pressure and blood lipid in both groups were remarkably improved (all P < 0.001) after treatment; the urea nitrogen and creatinine had no notable variation. CONCLUSION: Acupoint thread embedding combined with western medicine routine therapy can not only reduce the urinary micro-albumin excretion of type II early diabetic nephropathies, but also improve the Chinese medicine symptoms and the therapeutic effect is superior to that of simple western medicine routine therapy.

[Clinical observation on therapeutic effect and instant analgestic effect of inhibitory-needling at Ashi point as major point for treatment of piriformis syndrome].

OBJECTIVE: To observe therapeutic effect and analgesic eJfect of needling at Ashi point for treatment of piriformis syndrome. METHODS: Eighty cases were randomly divided into an inhibitory-needling method group (INM) and a routine-needling method group (RNM), forty cases in each group. The acupuncture at Ashi point as major point with inhibitory-needling munipulation was applied in the INM group, the acupuncture at Huantiao (GB 30) ,Zhibian (BL 54) ,Yanglingquan (GB 34) with routine-needling method in the RNM group. The therapeutic effect, the instant analgesic effect and score of Visual Analogue Scale (VAS) were observed. RESULTS: After treatment, the effective rate of 92.5% in the INM group was better than that of 82.5% in the RNM group (P < 0.05); the instant analgesic effect rate of 75.0% in the INM group was better than that of 52.5% in the RNM group (P < 0.01); the score of VAS in both groups had decreased with a significant difference between two groups (P < 0.05). CONCLUSION: The treatment of inserting the needle at Ashi point as major point with inhibitory-needling method has excellent effectiveness and good instant analgesic effect, with fewer points selection and rapid effect.

[Clinical observation on acupuncture for intervening the response in gastroscopy].

OBJECTIVE: To observe the clinical effect of acupuncture for controlling the adverse response in gastroscopy. METHODS: Ninety-seven cases of gastroscopy were randomly divided into an observation group of 52 cases and a control group of 45 cases. The observation group were treated by acupuncture at Hegu (LI 4), Neiguan (PC 6) and Zusanli (ST 36), in combined with oral administration of Lidocaine; the control group were treated by simple administration of Lidocaine. Changes of the adverse response, blood pressure and heart rate, and satisfactory degrees and the willing re-examination rate were investigated in the two groups. Results In the observation group, the nausea and vomiting, salivation, restlessness, breath holding and other adverse responses in gastroscopy were significantly decreased as compared with those in the control group (P < 0.01), and the blood pressure and heart rate were more stable than in the control group, and the satisfactory degree and willing re-examination rate were higher than the control group (P < 0.01). CONCLUSION: Acupuncture can effectively control the adverse response in gastroscopy.