hsa_circ_0140180 affects the migration and invasion of esophageal squamous cell carcinoma cells by regulating miR-1287-5p / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨hsa_circ_0140180在食管鳞状细胞癌（ESCC）细胞中的表达水平及对其细胞恶性生物学行为的影响与分子机制。方法：收集2018年11月至2019年3月间在南充市中心医院胸心外科手术切除的6对ESCC组织和对应癌旁组织并进行全转录组测序，筛选出在ESCC组织中低表达的hsa_circ_0140180；建立过表达hsa_circ_0140180的TE-1和KYSE30细胞，qPCR法检测hsa_circ_0140180在人正常食管上皮细胞、ESCC细胞中的表达，以及过表达hsa_circ_0140180后TE-1和KYSE30细胞中miR-1287-5p的表达；CCK-8法和FCM检测过表达hsa_circ_0140180对TE-1和KYSE30细胞增殖和周期的影响；划痕实验和Transwell实验检测过表达hsa_circ_0140180对TE-1和KYSE30细胞迁移和侵袭能力的影响，双荧光素酶报告实验验证hsa_circ_0140180与miR-1287-5p的靶向关系。WB法检测过表达hsa_circ_0140180对TE-1和KYSE30细胞中EMT相关蛋白的表达及PI3K-Akt通路的磷酸化水平的影响。结果：转录组测序和qPCR结果显示，hsa_circ_0140180在ESCC组织和细胞中呈低表达（P<0.05或P<0.01），并确认其闭合环状分子特征且定位于细胞质。过表达hsa_circ_0140180能明显抑制ESCC细胞的迁移及侵袭能力（P<0.05），但不影响其增殖和周期。双荧光素酶报告基因实验证实hsa_circ_0140180靶向结合miR-1287-5并负调控其表达（P<0.01）。过表达hsa_circ_0140180能显著上调TE-1和KYSE30细胞中E-cadherin的表达（（P<0.05），而显著下调Snail的表达（P<0.05）和PI3K-Akt通路的磷酸化水平（P<0.01或P<0.001）。结论：hsa_circ_0140180在ESCC细胞及组织中呈低表达，其可能通过调控miR-1287-5p表达来降低PI3K-Akt通路的磷酸化水平和抑制EMT进程，从而影响ESCC细胞的迁移及侵袭能力。

Effects of 1,25(OH)2D3 regulating ERK pathway on the biological behaviors of esophageal squamous cell carcinoma cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨1,25-二羟维生素D3 [1,25(OH)2D3]对食管鳞状细胞癌（esophageal squamous cell carcinoma，ESCC）细胞增殖、迁移和细胞周期的影响及其相关机制。方法：用不同浓度1,25(OH)2D3处理ESCC细胞TE-11、KYSE30、TE-1和KYSE510后，用CCK-8法检测细胞的增殖能力。再用浓度分别是0、0.1、0.15、0.2 μmol/L的1,25(OH)2D3处理TE-11和KYSE30细胞，划痕愈合实验、流式细胞术分别检测细胞的迁移能力和细胞周期分布情况，WB法检测细胞中cyclin D1、P27、ERK和p-ERK蛋白的表达水平。结果：1,25(OH)2D3显著抑制TE-11和KYSE30细胞的增殖能力，其抑制程度呈时间依赖性和浓度依赖性。0.1和0.2 μmol/L的1,25(OH)2D3处理48 h后，与空白对照组比较，TE-11和KYSE30细胞的迁移能力均显著降低（P<0.05或P<0.01），处于G0/G1期细胞显著增加（P<0.05或P<0.01），细胞中cyclin D1和p-ERK蛋白水平显著下调、P27蛋白水平明显上调（P<0.05或P<0.01）而ERK蛋白的表达无明显变化。结论：1,25(OH)2D3显著抑制ESCC细胞的增殖和迁移能力并阻滞细胞周期进程，其可能通过调控ERK信号通路而发挥作用。

TLR7 mediates increased vulnerability to ischemic acute kidney injury in diabetes

SUMMARY OBJECTIVE Diabetes is a risk factor for acute kidney injury (AKI). However, its mechanism of pathogenesis has not been elucidated. The aim of the study was to investigate the role of inflammation and the toll-like receptor 7 (TLR7) in ischemic AKI for diabetes. METHODS A high glucose hypoxia-reoxygenation model of human renal tubular epithelial (HK-2) cells was used to generate AKI induced by ischemia-reperfusion in diabetes. The activity of cells was measured by CCK-8 assay and LDH activity. Inflammatory cytokines were assessed by ELISA. TLR7, MyD88, and NF-κB expressions were examined by western blotting. Apoptosis was evaluated by flow cytometry. RESULTS The high glucose group and low glucose group were subjected to hypoxia-reoxygenation. The low glucose group developed only mild cell damage, apoptosis, and inflammatory response. In contrast, an equivalent hypoxia-reoxygenation injury provoked severe cell damage, apoptosis, and inflammatory response in the high glucose group. Expression of TLR7 and its related proteins were measured in the high glucose group before and after hypoxia-reoxygenation. The high glucose group exhibited more significant increases in TLR7 expression following hypoxia-reoxygenation than the low glucose group. In addition, the expression of TLR7 and its related proteins after hypoxia-reoxygenation were higher in the high glucose group than in the low glucose group. Inhibition of TLR7 provides significant protection against ischemic injury in diabetes. CONCLUSION Our results suggest that diabetes increases the vulnerability to ischemia-induced renal injury. This increased vulnerability originates from a heightened inflammatory response involving the TLR7 signal transduction pathway.

RESUMO OBJETIVO O diabetes é um fator de risco para a lesão renal aguda (LRA). No entanto, seu mecanismo de patogênese não foi elucidado. O objetivo do estudo foi investigar o papel da inflamação e do receptor Toll-like 7 (TLR7) na LRA isquêmica no diabetes. MÉTODOS Um modelo de hipóxia-reoxigenação de células epiteliais tubulares renais humanas (HK-2) na presença de concentrações altas de glicose foi utilizado para gerar LRA induzida por isquemia-reperfusão em diabetes. A atividade das células foi medida pelo ensaio Cell Counting Kit-8 (CCK-8) e pela atividade da lactato desidrogenase (LDH). As citocinas inflamatórias foram avaliadas por ensaio imunoenzimático (Elisa). A expressão de TLR7, do fator de diferenciação mieloide 88 (MyD88) e do fator de transcrição nuclear-κB (NF-κB) foi examinada por Western blotting. A apoptose foi avaliada por citometria de fluxo. RESULTADOS Os grupos glicose alta e glicose baixa foram submetidos à hipóxia-reoxigenação. O grupo de baixa glicose desenvolveu apenas danos celulares ligeiros, apoptose e uma resposta inflamatória. Em contraste, no grupo de alta glicose, uma lesão equivalente de hipóxia-reoxigenação provocou danos celulares graves, apoptose e uma resposta inflamatória. A expressão de TLR7 e suas proteínas relacionadas foi medida no grupo de alta glicose antes e após a hipóxia-reoxigenação. O grupo de alta glicose exibiu maiores aumentos na expressão de TLR7 após hipóxia-reoxigenação do que o grupo de baixa glicose. Além disso, a expressão de TLR7 e suas proteínas relacionadas após a hipóxia-reoxigenação foi maior no grupo com alto nível de glicose do que no grupo com baixo nível de glicose. A inibição do TLR7 fornece proteção significativa contra a lesão isquêmica no diabetes. CONCLUSÃO Nossos resultados sugerem que o diabetes aumenta a vulnerabilidade à lesão renal induzida por isquemia. Essa vulnerabilidade acrescida tem por origem uma resposta inflamatória aumentada envolvendo a via de transdução de sinal do TLR7.

Exogenous neuregulin-1 attenuates STZ-induced diabetic peripheral neuropathic pain in rats

Abstract Purpose: To investigate whether modulating NRG1 could attenuate diabetic neuropathic pain and analyze the underlying mechanism. Methods: Male SD rats were randomly divided into control group, diabetic group, NRG1 intervention group. After STZ-induced 2 weeks, NRG1 intervention daily for consecutive 7 days. 4 weeks after NRG1 intervention, both the mechanical withdrawal threshold and the morphological changes of the dorsal root ganglion and sural nerve were observed. Meanwhile, the expression of NGF, IL-1β, TNF-α in spinal cord were determined. Results: Compared with the diabetic group, NRG1 treatment improved the mechanical withdrawal threshold in diabetic rats, pathological changes of dorsal root ganglion and sural nerve were alleviated by NRG1 treatment with electron microscopy imagine. Moreover, compared with the control group, the expression of NGF was significantly decreased and the production of IL-1β, TNF-α were markedly induced in diabetic group. Furthermore, NRG1 treatment could normalized the above effect as compared to diabetic group. Conclusion: NRG1 exerted positive effects on the behavioral and pathological changes of rats with STZ-induced diabetic neuropathic pain, the underlying mechanism might be related to the promotion of NGF excretion and the inhibition of inflammatory cytokines excretion.

Ginsenoside metabolite compound K alleviate collegen-induced arthritis through impairing dendritic cells function / 中国药理学与毒理学杂志

OBJECTIVE Ginsenoside metabolite compound K (CK) is a degradation product of ginsenoside in the intestine by bacteria. The anti-inflammatory and immunomodulatory activities of CK have been reported. This study investigated whether CK exerted its immunoregulatory effect through modulation of dendritic cells (DCs) function. METHODS In vivo, severity of collegen-induced arthritis (CIA), T cells and DCs subsets, phenotype of DC were assayed by flow cytometry, CCL19 and CCL21 level in lymph nodes assayed by ELISA. In vitro, bone marrow-derived DCs from normal mice were matured with lipopolysaccharide and treated with CK for 48 h. In vivo, bone marrow-derived DCs were generated from CIA mice before and 2 weeks into CK treatment. DCs were analyzed for migration, phenotype and T- cell stimulatory capacity. RESULTS CK alleviated the severity of CIA, decreased pDCs and mo-DCs, increased na?ve T cells in CIA mice lymph nodes, and suppressed CCL21 expression in lymph nodes. CK suppressed DCs migration induced by CCL21 and T cells-stimulatory capability of DC, down-regulated LPS-induced expression of CD80, CD86, MHCII and CCR7 on DCs. CONCLUSION This study elucidated the novel immunomodulatory property of CK via impairing function of DCs in priming T cells activation. These results provide an interesting novel insight into the potential mechanism by which CK contribute to the restoration of immunoregulation in autoimmune conditions.

Therapeutic effect of low molecular weight chitosan containing sepia ink on ethanol-induced gastric ulcer in rats

ABSTRACT PURPOSE: To evaluate the role of low molecular chitosan containing sepia ink (LMCS) in ethanol-induced (5 ml/kg) gastric ulcer in rats. METHODS: Animals were divided into four groups (n = 12): normal group (Normal), negative control group (Con), experiment group (LMCS) and positive control Omeprazole group (OMZ). Gastric empty rate was detected in the first 7 days. Rats were sacrificed at 7, 14 and 21 day for histology and ELISA detections. RESULTS: Gastric empty was no significant differences among the groups (P > 0.05). Histological observation showed gastric mucosal LMCS treated had better healing effect. Hydroxyproline (Hyp) was significantly increased from 7 day (P < 0.05). LMCS significantly inhibited malondialdehyde (MDA) generation for lipid peroxidation from 7 day (P < 0.05). LMCS significantly promoted the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) at the earlier stage (P < 0.05). OMZ had the similar effects above. As for myeloperoxidase (MPO), LMCS significantly decreased and restored it to normal levels from 7 day (P < 0.05), it is earlier than OMZ which is from 14 day. CONCLUSION: LMCS can improve gastric mucosa tissue repair, exert significant influences on oxidative and antioxidant enzyme activities and neutrophil infiltration.