Background:The molecular regulators driving melanoma pathogenesis remain incompletely defined. Although plasma proteins show associations with melanoma, their causal relationships are unclear. Methods:We performed Mendelian randomization (MR) using proteomic data from the UK Biobank Pharma Proteomics Project and deCODE genetics to identify plasma proteins causally linked to melanoma. Colocalization, summary data-based MR, heterogeneity in dependent instruments, and MR-phenome-wide association study analyses identified druggable targets. Transcriptome profiling of melanoma tissues and single-cell functional validation assessed the target roles. Results:Poly(adenosine diphosphate-ribose) polymerase 1 (PARP1) was established as a causal risk factor and druggable target for melanoma. Elevated PARP1 expression in melanoma tissues correlated with poor patient survival. Functional analyses revealed that PARP1 coordinates protumorigenic functions with cell-specific roles. Conclusions:In conclusion, our results demonstrate that PARP1 is a key driver of melanoma pathogenesis and represents a potential specific molecular target for therapeutic intervention.

HOX transcript antisense RNA (HOTAIR) is associated with the growth and metastasis of many human tumors, but its biological roles in malignant melanoma remain unclear. In this study, we show that HOTAIR is overexpressed in melanoma tissues and cells, especially in metastatic melanoma. High HOTAIR levels correlate with poor prognosis in melanoma patients. We also determined that HOTAIR functions as a competing endogenous RNA (ceRNA) for miR-152-3p. miR-152-3p was decreased and acted as a tumor suppressor in melanoma, and c-MET was the functional target of miR-152-3p. Furthermore, HOTAIR promotes the growth and metastasis of melanoma cells by competitively binding miR-152-3p, which functionally liberates c-MET mRNA and results in the activation of the downstream PI3k/Akt/mTOR signaling pathway. We determined that HOTAIR acts as a ceRNA to promote malignant melanoma progression by sponging miR-152-3p. This finding elucidates a new mechanism for HOTAIR in melanoma development and provides a potential therapeutic target for melanoma patients.

Objectives:Long noncoding RNAs plays the important part in tumor biology. SNHG19 was found to be a new oncogenic lncRNA in some malignant tumors. However, the effect of SNHG19 in hepatocellular carcinoma has not been reported. Methods:The expression of SNHG19 in hepatocellular carcinoma tissues were detected by using quantitative Real-Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR). Melanoma cases from The Cancer Genome Atlas were included in this study. cell counting kit-8 assay, Transwell, and scratch wound assay were used to explore the role of SNHG19 in melanoma cells. Luciferase reporter assays and RNA pull-down assay were used to explore the molecular mechanism of SNHG19 in hepatocellular carcinoma. Results:Here, we found that SNHG19 level was upregulated in hepatocellular carcinoma. Hepatocellular carcinoma patients with high levels of SNHG19 have shorter Disease-Free Survival (RFS). SNHG19 promotes the Protein Tyrosine Phosphatase 4A3 expression by sponging miR-137 to liberate Protein Tyrosine Phosphatase 4A3 mRNA transcripts. SNHG19 enhances the development of hepatocellular carcinoma by affecting miR-137/Protein Tyrosine Phosphatase 4A3 axis. Conclusion:These results demonstrated the effect of SNHG19 in the occurrence and progression of hepatocellular carcinoma. SNHG19 may be used as the specific molecular target in patients with hepatocellular carcinoma.

BACKGROUND:Keloid represents a benign skin tumor with many cancer-like features. Extracellular vesicles (EVs) derived from human adipose-derived stem cells (hADSCs) play a role in cell migration of multiple diseases.AIMS:This study aimed to investigate the impact of hADSC-EVs on human keloid fibroblasts (HKFs).METHODS:hADSCs were cultured to the 3rd generation, and subsequently assessed for their osteogenic, adipogenic, and chondrogenic differentiative abilities using flow cytometry, alizarin red, oil red O, and alcian blue staining techniques. hADSC-EVs were isolated through ultracentrifugation and subsequently identified. HKFs at the 3rd generation were subjected to treatment with hADSC-EVs to observe their endocytosis of EVs by immunofluorescence. CCK-8, wound healing, and Transwell assays were performed to test HKF proliferation and migration. The levels of autophagy proteins, collagens, and Janus kinase 2 (JAK2) and Signal Transducer and Activator of Transcription 3 (STAT3) were determined through Western blot analysis. Suppressor of cytokine signaling 1 (SOCS1) expression was determined by RT-qPCR and Western blot.RESULTS:hADSC-EVs were successfully isolated from hADSCs. PKH67-labeled hADSC-EVs were observed to be endocytosed by HKFs, resulting the inhibition of HKF proliferation, migration, as well as a reduction in collagen deposition. hADSC-EVs carried SOCS1 into HKFs to suppress HKF autophagy. SOCS1 downregulation in hADSC-EVs partially nullified the inhibitory effect of hADSC-EVs on HKFs. hADSC-EV-carried SOCS1 inhibited the activation of the JAK2/STAT3 pathway. JAK2/STAT3 pathway activation partially abrogated the suppression of hADSC-EVs on the proliferation, migration, and collagen deposition of HKF.CONCLUSION:hADSC-EVs carried SOCS1 into HKFs and suppressed HKF autophagy, proliferation, migration, and collagen deposition by inactivating the JAK2/STAT3 pathway.

Traumatic brain injury (TBI) is one of the important risk factors for the development of Alzheimer’s disease (AD). However, the molecular mechanism by which TBI promotes the progression of AD is not elucidated. In this study, we showed that the abnormal production of E2F1 is a major factor in promoting the neuropathological and cognitive deterioration of AD post-TBI. We found that repeated mild TBI can aggravate the neuropathology of AD in APP/PS1 mice. At the same time, the co-expression of E2F1 and beta-site APP cleaving enzyme 1 (BACE1) was upregulated when the mouse hippocampus was dissected. BACE1 is recognized as a rate-limiting enzyme for the production of Aβ. Here, we speculate that E2F1 may play a role in promoting BACE1 expression in AD. Therefore, we collected peripheral blood from patients with AD. Interestingly, there is a positive correlation between E2F1 and brain-derived neurotrophic factor–antisense (BDNF-AS), whereas BDNF-AS in AD can promote the expression of BACE1 and exhibit a neurotoxic effect. We established a cell model and found a regulatory relationship between E2F1 and BDNF-AS. Therefore, based on our results, we concluded that E2F1 regulates BDNF-AS, promotes the expression of BACE1, and affects the progression of AD. Furthermore, E2F1 mediates the TBI-induced neurotoxicity of AD.

Exosomes derived from neighboring v-raf murine sarcoma viral oncogene homolog B1 inhibitor (BRAFi)-resistant melanoma cells mediate the formation of resistance in melanoma cells sensitive to BRAFi. The function and molecular mechanisms of exosomal miRNA in BRAFi resistance of melanoma have not been studied. We found that the expression of miR-19a in BRAFi resistant melanoma cells was significantly higher than that in sensitive cells, and miR-19a contributes to the resistance of melanoma cells to BRAFi by targeting immunoglobulin-like domains protein 1 (LRIG1). miR-19a was highly enriched in exosomes secreted from BRAFi resistant melanoma cells, and these exosomal miR-19a promote the spread of BRAFi resistant. The reactivation of Protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) pathways is the main reason for the BRAFi resistant of melanoma cells. We demonstrated that exosomal miR-19a derived from melanoma cell promotes the formation and spread of BRAFi resistant in melanoma through targeting LRIG1 to reactivate AKT and MAPK pathway. Therefore, miR-19a may serve as a potential therapeutic target in melanoma patients with acquired drug resistance.

INTRODUCTION:Bone marrow mesenchymal stem cell-derived exosomes (BMSC-exos) may function as novel candidates for treating diabetic wounds due to their ability to promote angiogenesis. MATERIALS AND METHODS:This study investigated the effects of BMSC-exos on the growth and metastasis of human umbilical vein endothelial cells (HUVECs) treated with high glucose (HG). The exosomes were separated from BMSCs and identified. The cell phenotype was detected by 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and 5-ethynyl-2'-deoxyuridine, wound healing, and transwell assays, while the number of tubes was measured via tube formation assay. RESULT:The RNA and protein expression levels were studied using reverse transcription-quantitative polymerase chain reaction and western blotting, whereas integration of microRNA-99b-5p (miR-99b-5p) with THAP domain containing 2 (THAP2) was confirmed via dual-luciferase reporter and RNA pull-down assays. Results of transmission electron microscopy, nanoparticle tracking analysis, and laser scanning confocal microscopy revealed that exosomes were successfully separated from BMSCs and endocytosed into the cytoplasm by HUVECs. Similarly, BMSC-exos were found to promote the growth of HG-treated HUVECs, while their growth was inhibited by suppressing miR-99b-5p. THAP2 was found to bind to miR-99b-5p, where THAP2 inhibition reversed the miR-99b-5p-induced effects on cell growth, migration, and tube numbers. CONCLUSION:In conclusion, miR-99b-5p in BMSC-exo protects HUVECs by negatively regulating THAP2 expression.

Background There are a lot of studies on the treatment of tumors with hyperbaric oxygen, while most of them are in breast cancer, prostate cancer and so on. However, there are still few studies on hyperbaric oxygen in treating hepatocellular carcinoma (HCC). According to the current data, hyperbaric oxygen is an effective means to intervene in tumors. The Warburg effect is a unique marker of glucose metabolism in tumors related to hypoxia, making it possible for hyperbaric oxygen to interfere with the tumor through the Warburg effect.Method We used the hypoxia/hyperbaric oxygen(HBO)-exposed HCC cells for in vitro studies. Glucose uptake, lactic acid, and adenosine triphosphate (ATP) assessed the Warburg effect. The expression of miR-103a-3p in HCC was detected by using qRT-PCR. The effect of miR-103a-3p/TRIM35 expression level on the cells was measured using the CCK8 method and flow cytometry. The molecular biological mechanism of miR-103a-3p in HCC was examined using the luciferase reporter, MS2-RIP assays.Result HBO inhibited the Warburg effect in hypoxic HCC cells. HBO suppressed the expression of miR-103a-3p in hypoxic HCC cells, and miR-103a-3p inhibited the expression of TRIM35 in hypoxic HCC cells. With HBO exposure, miR-103a-3p/TRIM35 regulated the Warburg effect of hypoxic HCC cells.Conclusion These findings reveal that HBO regulates the Warburg effect of hypoxic HCC cells through miR-103a-3p/TRIM35 and inhibits tumor growth.

miR-224-5p has been shown to play both an oncogene and tumour suppressor role in many human tumours. However, the role and molecular mechanism of miR-224-5p in cutaneous melanoma remains unclear. miR-224-5p levels were downregulated in melanoma tissue, and low miR-224-5p expression was an independent risk factor for melanoma patients. miR-224-5p blocked proliferation, epithelial-to-mesenchymal transition (EMT), invasion, migration in BRAF wild-type melanoma cell, and overcome acquired BRAFi resistance in VMF-resistant melanoma cells. miR-224-5p exerted its role by directly repressing PAK4 to block the downstream CRAF/MEK/ERK pathways. We demonstrated that miR-224-5p inhibited melanoma growth and metastasis in vivo though xenograft tumor and pulmonary metastasis assay. Thus, miR-224-5p/PAK4-mediated CRAF/MEK/ERK pathways have therapeutic potential in melanoma treatment.

目的 探讨皮下及真皮层超减张缝合联合减张胶带外固定对胸腹部增生性瘢痕的临床应用效果.方法 回顾性分析自 2017 年 8 月至 2022年 8 月,南通大学附属丹阳医院烧伤整形外科 65 例胸腹部增生性瘢痕患者资料.将患者按治疗方法分为治疗组(超减张缝合联合减张胶带,35 例)和对照组(传统减张缝合,30 例),比较两组患者术后 6个月的温哥华评分量表结果以及瘢痕宽度.结果 所有患者术后均一期愈合,未出现严重并发症.随访 6 个月,两组温哥华评分量表评分比较,治疗组低于对照组,差异具有统计学意义(P<0.05);两组瘢痕宽度比较,治疗组明显窄细,差异具有统计学意义(P<0.05).结论 采用分层超减张缝合联合减张胶带能有效地降低切缘机械张力,并在较长时间维持减张作用,术后切口瘢痕增生不明显.

目的:研究银离子敷料覆盖联合负压吸引(Vacuum-assisted closure,VAC)在慢性难愈性创面修复患者中的应用效果.方法:选择笔者医院2016年3月-2020年9月诊治的98例慢性难愈性创面患者,随机数表法分为对照组(49例)和研究组(49例),前者给予传统治疗,后者给予银离子敷料覆盖联合VAC治疗.比较两组患者治疗前后创面感染、组织再上皮化、创面瘢痕情况变化,并评估疼痛程度,统计敷料更换和创面恢复情况.结果:治疗前两组患者细菌培养阳性率、C反应蛋白(C-reactive protein,CRP)、基质金属蛋白酶2(Matrix metalloproteinase-2,MMP-2)以及红细胞沉降率(ESR)水平、瘢痕评分比较无显著差异(P>0.05),治疗后研究组上述指标水平均降低,且低于对照组,差异有统计学意义(P<0.05);治疗前两组患者转化生长因子β1(TGF-β1)和血管内皮生长因子(VEGF)水平比较无显著差异(P>0.05),治疗后研究组上述指标水平均升高,且高于对照组,差异有统计学意义(P<0.05);治疗前两组患者疼痛评分比较无显著性差异(P>0.05),治疗后7 d、14 d、1个月研究组评分明显降低,且低于对照组,差异有统计学意义(P<0.05);研究组患者敷料更换次数、创面肉芽组织生长、上皮形成以及愈合时间明显低于对照组,差异有统计学意义(P<0.05).结论:银离子敷料覆盖联合VAC技术可较好的控制创面感染、促进组织再上皮化的形成,同时有利于减轻疼痛和瘢痕,促进创面更好的愈合恢复.

Background: Long non-coding RNAs (lncRNAs) play an key role in the biological processes of various malignant tumors. SNHG16 has been confirmed to be associated with the progression of various cancers. However, function and molecular mechanism of SNHG16 in melanoma have not been studied by scholars. Methods: The expression of SNHG16 in melanoma tissues were detected by using qRT-PCR. Melanoma cases from The Cancer Genome Atlas (TCGA) and GEO#GSE15605 were included in this study. CCK-8 assay, EdU assay, transwell and scratch wound assay were used to explore the role of SNHG16 in melanoma cells. Luciferase reporter assays and RNA pull-down assay were used to explore the molecular mechanism of SNHG16 in melanoma. Results: Here, we found that SNHG16 was up-regulated in melanoma. SNHG16 enhances the growth and metastasis of melanoma. SNHG16 can promote the expression of P21-activated kinases 2 (PAK2) by sponging miR-205-5p. PAK2 is the target gene of miR-205-5p. We demonstrated that SNHG16 promotes the metastasis and growth of melanoma through miR-205-5p/PAK2 axis. Conclusion: This study firstly confirmed the role and mechanism of SNHG16 in the occurrence and development of melanoma. Therefore, SNHG16 may become a key point in the diagnosis, prognosis and treatment of melanoma patients in the future.

Objective:To investigate the efficacy of ultrasound-guided thoracic spinal nerve root block combined with radial shock wave therapy in elderly patients with chest and back postherpetic neuralgia (PHN).Methods:Sixty patients with PHN, aged≥60 years old, both sexes, with a duration of 1-6 months and numerical rating scale (NRS)>6, were collected from August 2019 to December 2020 in the Department of Pain Medicine of Zhenjiang First People's Hospital, and were randomly divided into shock wave group and combined nerve block group by random number table method, 30 cases in each group. Patients were received radial shock wave therapy in the shock wave group, while received radial shock wave therapy combined with ultrasound-guided thoracic spinal nerve root block in the combined nerve block group, once/week for a total of 4 times; and all patients were supplemented with basic drug therapy. NRS, Pittsburgh sleep quality index (PSQI), Hamilton anxiety scale (HAMA), quality of life index (QL-index), overall efficacy and related complications were evaluated before the treatment and 1, 4, 8 and 12 weeks after the treatment.Results:There were no differences in NRS, PSQI, HAMA, and QL-index scores before the treatment between the two groups (all P>0.05). At 1, 4, 8 and 12 weeks after the treatment, NRS, PSQI and HAMA scores were significantly lower and the QL-index score was significantly higher than those before the treatment in both groups (all P<0.05). NRS, PSQI, HAMA were significantly lower, and QL-index score was higher at each time point in the combined nerve block group than those in the shock wave group (all P<0.05). At 12 weeks after the treatment, the excellent rate was higher in the combined nerve block group than that in shock wave group [90.0% (27/30 cases) vs. 63.3% (19/30 cases), P<0.05]. Conclusion:Ultrasound-guided thoracic spinal nerve root block combined with radial shock wave therapy can effectively relieve clinical symptoms and improve the quality of life in elderly patients with thoracic PHN, and the efficacy is superior to shock wave only.

MicroRNAs (miRNAs) are small endogenous noncoding RNAs that regulate genome expression posttranscriptionally and are involved in autoimmune diseases. Previous studies have indicated that follicular helper T (Tfh) cells play a critical role in the pathogenesis of Graves’ disease (GD). However, the molecular mechanisms that contribute to circulating Tfh memory cell response in GD patients remain incompletely understood. This study aimed to investigate the role of miRNAs on circulating Tfh memory cells in GD patients. Herein, our data showed that the proportion of circulating Tfh memory cells, the transcript levels of IL-21, and the plasma concentrations of IL-21 were increased in the peripheral blood from GD patients. We also found that inducible co-stimulator (ICOS) expression, an important molecule expressed on Tfh cells, were significantly augmented in the peripheral blood mononuclear cells (PBMCs) from GD patients and positively correlated with the percentage of circulating Tfh memory cells and the transcript levels of IL-21 in GD. Intriguingly, miRNA sequencing screened miR-29a-3p expression was downregulated and inversely correlated with ICOS expression and the frequency of circulating Tfh memory cells in patients with GD. Luciferase assay demonstrated that ICOS was the direct target gene of miR-29a-3p, and miR-29a-3p could inhibit ICOS at both transcriptional and translational levels. Overexpression of miR-29a-3p reduced the proportion of circulating Tfh memory cells. Moreover, miR-29a-3p expression negatively correlated with serum concentrations of TSH receptor antibody (TRAb) in GD patients. Collectively, our results demonstrate that miR-29a-3p emerges as a post-transcriptional brake to limit circulating Tfh memory cell response in GD patients and may be involved in the pathogenesis of GD.

Introduction: The long non-coding RNA Brain-derived nutritional factor anti-sense RNA (BDNF-AS) is a type of anti-sense RNA that has been proven to play a crucial role in the occurrence and development of certain nervous system disorders. However, the role and molecular mechanism of BDNF-AS in Alzheimer's disease (AD) have not been elucidated yet. Methods: Peripheral blood samples were collected from outpatients with AD as well as from normal elderly individuals in the community, and the expression of BDNF-AS was analysed using quantitative reverse transcription-polymerase chain reaction. An in vitro model was constructed, and the effect of BDNF-AS expression level on the cells was measured using the CCK8 method and flow cytometry. The molecular biological mechanism of BDNF-AS in AD was examined using the luciferase reporter, MS2-RIP, and RNA pulldown assays. Result: We found that the expression of BDNF-AS was elevated in the peripheral blood of patients with AD and that increased BDNF-AS expression may be associated with the cognitive status of such patients. The results confirmed that BDNF-AS could promote neurotoxicity in the in vitro model. Then, we uncovered that BDNF-AS promotes the expression of BACE1 through the competitive binding of miR-9-5p, thereby promoting amyloid deposition. Finally, through the Morris water maze, we found that the high expression of BDNF-AS promoted cognitive impairment in AD mice. Conclusion: The obtained results suggest that BDNF-AS plays a crucial role in the occurrence and development of AD. As a new pathogenic gene of AD, BDNF-AS may be used as a therapeutic target or as a prognostic marker in patients with AD.

目的 探讨负压创面治疗联合高压氧治疗糖尿病足的临床疗效.方法 采用回顾性病例对照研究分析江苏大学附属人民医院整形烧伤科自2013年2月至2017年12月收治的46例糖尿病足患者的临床资料,根据治疗方法分为对照组(24例)和联合治疗组(22例).糖尿病足创面按Wagner分级为3～5级;共46足.对照组采用控制血糖、抗感染、清创、创面湿性换药等方法,根据创面情况予以换药、植皮或截肢治疗.联合治疗组在对照组治疗方法基础上采用负压创面治疗联合高压氧治疗,并根据创面情况予以换药、植皮、放置负压引流材料或截肢治疗.治疗后观察患者创面愈合、住院时间及截肢情况.结果 治疗总有效率:联合治疗组(59％)明显高于对照组(33％).对照组和联合治疗组的住院时间分别为(25.2±16.5)d和(28.7±12.8)d.对照组中9例患者共截趾11趾,大腿截肢7例,小腿截肢1例.联合治疗组中5例患者共截趾8趾,小腿截肢2例.与对照组相比,联合治疗组截肢率明显降低(z=-2.617,P<0.05),且踝关节以上截肢率明显下降(33％:9％,P<0.05).结论 采用负压创面治疗联合高压氧治疗糖尿病足患者,可显著降低截肢率,改善患者出院后的生活质量.

Background Cancer-secreted exosomal miRNAs regulates the biological processes of many tumours. The serum level of exosomal miR-106b-5p is significantly increased in melanoma patients. However, the role and molecular mechanisms of exosomal miR-106b-5p in melanoma remains unclear. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-106b-5p and EphA4 in melanoma tissues. Transmission electron microscopy (TEM) and western blotting were used to identify exosome. QRT-qPCR and Cy3-labelled miR-106b-5p were used to demonstrated the transmission of melanoma cell-secreted exosomal miR-106b-5p. Western blotting, Immunofluorescence, adhesion, transwell and scratch wound assay were used to explore the role of exosomal miR-106b-5p in melanocytes. Luciferase reporter assays and RNA-Chromatin Immunoprecipitation (ChIP) assay were used to confirm whether erythropoietin-producing hepatocellular carcinoma receptor A4 (EphA4) was a direct target of miR-106b-5p. Results We found that miR-106b-5p levels were increased in melanoma tissue, and high miR-106b-5p expression is an independent risk factor for the overall survival of patients with melanoma. miR-106b-5p is enriched in melanoma cell-secreted exosomes and transferred to melanocytes. Exosomal miR-106b-5p promotes the epithelial-to-mesenchymal transition (EMT), migration, invasion and adhesion of melanocytes. Exosomal miR-106b-5p exerted its role by targeting EphA4 to activate the ERK pathway. We demonstrated that exosomal miR-106b-5p promoted melanoma metastasis in vivo through pulmonary metastasis assay. Conclusions Thus, melanoma cell-secreted exosomal miR-106b-5p may serve as a diagnostic indicator and potential therapeutic target in melanoma patients.

Objective:To investigate the role and mechanism of metformin in algesia of rats with type 2 diabetic neuropathic pain (DNP).Methods:Eighty sprague-dawley rats were randomly divided into normal control group ( n=15) and high-fat and high-glucose group ( n=65); normal diet and high-fat and high-sugar diet were given, respectively; before and 8 weeks after feeding, the body mass of rats and fasting blood glucose level were recorded, fasting insulin level was detected by ELISA, and insulin sensitivity index (ISI) was calculated. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) 8 weeks after feeding (baseline values) were measured in the high-fat and high-glucose group; after 12 h of fasting, intraperitoneal injection of streptozotocin (STZ, 35 mg/kg) was performed; 3 d after fasting, blood glucose was measured; 14 d after STZ injection, body mass was recorded and MWT and TWL were measured again: when MWT and TWL were ≤85% baseline values, it was defined that DNP model was successfully established ( n=45); and the left were into the diabetic painless group ( n=15). The rats with successful DNP were randomly divided into DNP group, DNP+vehicle group and DNP+metformin group ( n=15); 14 d after STZ injection, rats in the DNP+metformin group were given intraperitoneal injection of metformin (200 mg/kg) once daily for 14 consecutive d; DNP group did not accept any treatment, and rats in DNP+vehicle group were intraperitoneally injected with same amount of normal saline. MWT and TWL of all rats were measured 14 d after STZ injection, and 3, 7, 14 and 21 d after metformin injection. The expression levels of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α were detected by ELISA 7, 14 and 21 d after metformin injection. The fluorescence intensity of ionized calcium binding adaptor molecule-1 (Iba-1) in the spinal cord was detected by immunofluorescence staining, and the expression levels of Toll-like receptor 4 (TLR4), nuclear transcription factor (NF)-κB, phosphorylated (p)-NF-κB, adenylate activated protein kinase (AMPK), p-AMPK, and peroxisome proliferator activated receptor-γ coactivator (PGC)-1α in the spinal cord were detected by Western blotting 21 d after metformin injection. Results:(1) After 8 weeks of feeding, the body mass of rats in the high-fat and high-glucose group was significantly higher than that in the normal control group ( P<0.05); and the body mass of rats in the high-fat and high-glucose group was statistically lower than that in the normal control group 14 d after STZ injection ( P<0.05). Three d after STZ injection, the blood glucose level in high-fat and high-glucose group was significantly higher than that in normal control group ( P<0.05). After 8 weeks of feeding, the insulin level of high-fat and high-glucose group was statistically higher than that of normal control group, and the ISI in the high-fat and high-glucose group was significantly decreased as compared with that in the normal control group ( P<0.05). (2) As compared with those in the normal control group and diabetic painless group, MWT and TWL of DNP group and DNP+vehicle group were significantly decreased at each time point ( P<0.05). Three, 7, 14 and 21 d after metformin injection, MWT and TWL in DNP+metformin group were significantly increased as compared with those in DNP group and DNP+vehicle group ( P<0.05). (3) Seven, 14, and 21 d after metformin injection, the levels of IL-6, IL-1β and TNF-α in the spinal cord of rats in the DNP group and DNP+vehicle group were significantly increased as compared with those in the normal control group and diabetic painless group ( P<0.05); as compared with those in the DNP group and DNP+vehicle group, the levels of IL-6, IL-1β and TNF-α in the spinal cord of DNP+metformin group were significantly decreased ( P<0.05). (4) As compared with normal control group and diabetic painless group, the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while the fluorescence intensity of Iba-1 and number of Iba-1 positive cells in spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+ vehicle group ( P<0.05). (5) As compared with those in the normal control group and diabetic painless group, the TLR4 and p-NF-κB protein expressions and p-NF-κB/NF-κB values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly increased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly decreased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). As compared with those in the normal control group and diabetic painless group, the PGC-1α protein expression and p-AMPK/AMPK values in the spinal cord tissues of DNP group and DNP+vehicle group were significantly decreased ( P<0.05); while those in the spinal cord tissues of DNP+metformin group were significantly increased as compared with those in the DNP group and DNP+vehicle group ( P<0.05). Conclusion:Metformin, by activating AMPK/PGC-1α signaling pathway, may inhibit the TLR4/NF-κB expression, reduce the activation of microglia and the expressions of pro-inflammatory factors, and thus alleviate DNP.

The lncRNA ZFAS1 plays a carcinogenic regulatory role in many human tumours, but it is rarely reported in pancreatic cancer. We identify the role and molecular mechanisms of ZFAS1 in pancreatic cancer. The expression of ZFAS1, miR-497-5p and HMGA2 in pancreatic cancer tissues was detected by qRT-PCR. Pancreatic cancer data in The Cancer Genome Atlas were also included in this study. CCK8, EdU, transwell and scratch wound assays were used to investigate the biological effects of ZFAS1 in pancreatic cancer cells. MS2-RIP, RNA pull-down, RNA-ChIP and luciferase reporter assays were used to clarify the molecular biological mechanisms of ZFAS1 in pancreatic cancer. The role of ZFAS1 in vivo was also confirmed via xenograft experiments. ZFAS1 was overexpressed in pancreatic cancer tissues. ZFAS1 promoted the growth and metastasis of pancreatic cancer cells, and miR-497-5p acted as a tumour suppressor gene in pancreatic cancer by targeting HMGA2. We also demonstrated that ZFAS1 exerts its effects by promoting HMGA2 expression through decoying miR-497-5p. We also found that ZFAS1 promoted the progression of pancreatic cancer in vivo by modulating the miR-497-5p/HMGA2 axis. In conclusion, this study revealed a new role for and the molecular mechanisms of ZFAS1 in pancreatic cancer, identifying ZFAS1 as a novel target for the diagnosis and treatment of pancreatic cancer.