OBJECTIVE: To explore the effects of nandrolone phenylpropionate (NP) on the expression level of pro alpha 1 (I) collagen after burn in rats and the possible mechanism involved in the process. METHODS: Thirty-two Wistar rats with a deep second-degree scald injury and 20% of total body surface area were randomly divided into two groups to receive either 5 mg/kg NP(NP group) or normal saline (control group) every other day. We analyzed the mean integrated optical density(mIOD) of androgen receptor (AR) to determine the distribution and expression of AR in fibroblasts by immunohistochemistry, and measured expression level of pro alpha 1 (I) collagen mRNA by quantitative fluorescent RT-PCR to find the relation between expressions of AR and pro alpha 1 (I) collagen mRNA. The total specimens were obtained from the scalded rats after 4, 7, 14 and 21 of after burn. RESULTS: The expression of pro alpha 1 (I) collagen mRNA in NP group was significantly higher than that in control group on the 7th, 14th and 21st days(P lt; 0.05), but there was no significant difference on the 4th day. The density of AR in fibroblasts had significant difference (P lt; 0.05) between the two groups after 4, 7, 14 and 21 days. A positive relationship existed between the expression of pro alpha 1 (I) collagen mRNA and quantity of AR in fibroblasts(r = 0.836). CONCLUSION: The nandrolone phenylpropionate increased the expression of pro alpha 1 (I) collagen mRNA and enhanced the density of AR in fibroblasts. The higher expression of pro alpha 1 (I) collagen mRNA had a relation with the change of quantity of AR in fibroblasts.
Objective To systematically evaluate the correlation between the expression of microRNA (miRNA)-21 and the prognosis of esophageal cancer. Methods PubMed, Cochrane Library, Embase, Wanfang Data, China National Knowledge Infrastructure and VIP Databases were searched by for the literature on the correlation between miRNA-21 and the prognosis of esophageal cancer till July 10, 2022. Two researchers independently performed literature screening, quality evaluation, and data extraction. Statistical analysis was conducted with Stata 14.0. Results A total of 13 articles were included, including 1 204 patients. The results of meta-analysis showed that: the overall survival (OS) of patients with high expression of miRNA-21 was lower than that of patients with low expression of miRNA-21 [hazard ratio (HR)=2.11, 95% confidence interval (CI) (1.56, 2.84), P<0.001]. miRNA-21 expression was not associated with disease free survival [HR=2.53, 95%CI (0.67, 8.22), P=0.182]. The OS of Asian patients with high expression of miRNA-21 was significantly lower [HR=2.44, 95%CI (1.71, 3.49), P=0.005], while the OS of non-Asian patients was not related to miRNA-21 expression [HR=1.34, 95%CI (0.94, 1.91), P=0.363]. The high expression of miRNA-21 was correlated with the decreased OS in patients with esophageal squamous cell carcinoma [HR=2.22, 95%CI (1.52, 3.26), P=0.001], while the OS in patients with esophageal adenocarcinoma was not correlated with the expression of miRNA-21 [HR=1.39, 95%CI (0.63, 3.06), P=0.409]. Conclusion The overexpression of miRNA-21 is associated with poor prognosis and might be regarded as a potential prognostic biomarker for patients with esophageal cancer.
Objective To construct the expression short hairpin RNA （shRNA） targeting gene kir2. 1 in rat myocardial cells, named pEGFP6 kir2. 1, and to observe the effects on the expression of messenger RNA（mRNA） and protein of gene kir2. 1 as well as the changes of myocardial beating rates. Methods Five RNA interference （RNAi） sites targeting the rat kir2. 1 gene was selected, designed and synthesized five pairs of oligonucleotides fragments ,annealed them to double-strand, then cloned them into the vectors containing U6 promoter,obtained the vector expressing five aim genes. Rat myocardial cells were divided into three groups： Experimental group, negative plasmid control group and normal control group. Reverse transcription-polymerase chain reaction（RT PCR） and Western-blot were carried out to detect the expression of the mRNA and protein of gene kir2.1 and the beating rates of myocardial cells were observed after 72 h. Results The expression of mRNA and protein of gene kir2. 1 of experimental group were markedly lower than that of other two control groups after 72 h（P〈0.01）. There was no statistically significant between two control groups. The beating rate in experimental group was much faster than other two control groups （P〈0.01）, remained unchanged in both negative plasmid control group and normal control group. Conclusion Plasmid pEGFP6-kir2.1 could suppress the expression of the mRNA and protein of kir2.1 and increase the rat cardiac muscle cell beats.
Objective To construct vectors that express phosphatidylinositol-3-kinase, catalytic, beta polypeptide （PIK3cb） shRNA in eukaryon plasmid catalyzed by PI3K in rat, then test their effects on intimal hyperplasia in transplanted vein graft. Methods One hundred and fifty SD rats were randomly divided into six groups （n=25, in each group）: blank (25% Pluronic F-127), shRNA-1, shRNA-2, 1/2 （shRNA-1+shRNA-2）, negative control (pGenesil-1 scramble shRNA) and positive control （wortmannin） group. The jugular vein in rats were interpositioned autologously into the common carotid artery. shRNA and 25% Pluronic F-127 were mixed and coated around the transplanted vein in three PIK3cb shRNA groups. Every 5 samples were removed according to the time point (1, 3, 7, 14 and 28 days after operation), respectively. The thickness of intima and neointima area were calculated and analyzed by computer system. The PCNA expression was detected by Western blot and SP immunohistochemistry. Results The intimal thickness of three PIK3cb shRNA groups were lower than those in the blank group and negative control group on day 3, 7, 14, 28 after operation (P＜0.05); The neointima area in three PIK3cb shRNA groups (except shRNA-2 group on day 3, 7) began to decrease significantly from day one (P＜0.05). The protein expression of PCNA in three PIK3cb shRNA groups on day 3 after operation were decreased compared with blank group and negative group (P＜0.05). The percentage of the PCNA positive cells area in three PIK3cb shRNA groups were significantly lower than those in blank group and negative control group in each time point (Plt;0.05). There were no significant differences between blank and negative control group in different time points (Pgt;0.05). Conclusion The PIK3cb shRNA can effectively inhibit the proliferation of vascular smooth muscle cell, which may provide a new gene therapy for the prevention of vein graft restenosis after bypass grafting.
ObjectiveTo study the expressions of microRNA-221 (miR-221) and the protein of phosphatase and tension protein homologue (PTEN) in the proximal and distal stumps after sciatic nerve injury in rats and their correlation with the repair of peripheral nerve injury, so as to provide a new target for clinical diagnosis of peripheral nerve injury.MethodsNinety-six male Sprague-Dawley rats of SPF grade were selected to establish sciatic nerve injury models. Twenty-four rats were sacrificed at 0 (immediately after operation), 1, 4, and 7 days after operation. The proximal and distal sciatic nerve fragments were taken under aseptic conditions. The expression of miR-221 was detected by real-time fluorescent quantitative PCR, and the expression of PTEN protein was detected by Western blot and immunofluorescent staining. The relationship between miR-221 and PTEN was verified by dual-luciferase reporter gene. At the same time, the ultrastructure of nerve stump was observed by transmission electron microscopy.ResultsThe results of real-time fluorescent quantitative PCR, Western blot, and immunofluorescence staining showed that the relative expression of miR-221 in the proximal and distal stumps increased gradually with time, and the relative expression of PTEN protein decreased gradually, and the differences between different time points after operation were significant (P<0.05). At 1, 4, and 7 days after operation, the relative expression of miR-221 in proximal stump was significantly higher than that in distal stump, and the relative expression of PTEN protein in proximal stump was significantly lower than that in distal stump (P<0.05). Dual-luciferase reporter gene suggested that PTEN was the target for miR-221. Transmission electron microscopy observation showed that the normal morphological structure was observed at 0 day after operation, and the proliferation of Schwann cells and degeneration of axons and myelin sheaths gradually increased with time. There was no significant difference between proximal and distal stumps at 1 day after operation. At 4 and 7 days, Schwann cells proliferated more in proximal stump than in distal stump, and the degeneration of axons and myelin sheaths was less.ConclusionAfter sciatic nerve injury in rats, the up-regulation of the miR-221 expression targets the down-regulation of PTEN expression, which results in the difference of expression levels of miR-221 and PTEN in proximal and distal stumps. This phenomenon may play a role in promoting nerve repair after peripheral nerve injury.
ObjectiveTo investigate the changes of lung function after exposure to fine particulate matter (PM2.5) for 60 days and the expression of miR-146 in mice.MethodsThirty SPF BALB/c mice were treated with noninvasive tracheal instillation of fine particulate matter suspension at different doses (2.5 mg/kg, 5.0 mg/kg, 10.0 mg/kg) for 2 months (two times one week), the blank group and normal saline group were set as control groups. The mice were examined and killed on the next day after the last instillation. Histopathological changes of the lungs, pro-infammatory factors levels in the lung tissues, pulmonary functions and the relative expression of miR-146a and miR-146b in the lung tissues were detected.ResultsPeak inspiratory flow (PIF) and peak expiratory flow (PEF) were decreased significantly after PM2.5 exposure, however, lung resistance increased and maximal voluntary ventilation reduced from the general tendency without significant difference. Hematoxylin-eosin stain showed lymphocyte infiltration and macrophage infiltration by phagocytic particles, alveolar spacer widening, inflammatory response increased with the increase of PM2.5 exposure dosage. Pro-infammatory factors as interleukin-6 in the bronchoalveolar lavage fluid, interferon-γ and tumor necrosis factor-α in the lung homogenate were increased significantly by enzyme linked immunosorbent assay. The relative expressions of miR-146a and miR-146b were up-regulated remarkablely in treatment groups compared to the control group by real-time fluorescence quantitative polymerase chain reaction, which had negative relationships with PIF and PEF.ConclusionsThe lung function of mice decreases significantly after exposure to fine particulate matter, and the expression of miR-146 is up-regulated.
ObjectiveTo investigate the role of non-coding RNA (ncRNA) in the proliferation, migration and metastasis of hepatocellular carcinoma (HCC) and the mechanism of HCC resistance to sorafenib.MethodThe literatures of ncRNA studies related to the incidence of HCC in recent years were reviewed, and the relationship between different ncRNAs and the proliferation, migration and metastasis of HCC was summarized, and the mechanism of sorafenib resistance in the HCC was analyzed from the perspective of ncRNA.ResultsThere were many kinds of ncRNAs, which were classified into the long ncRNA and short ncRNA according to their length. Currently, microRNA, which was widely studied, belonged to the short ncRNA. The regulation of the expressions of different microRNAs and long ncRNA could enhance or inhibit the signaling pathway of the producing HCC and played an important guiding role in the diagnosis and treatment of HCC. Meanwhile, the targeted regulation of this ncRNA could reverse the sorafenib resistance in the HCC.ConclusionsncRNA plays an important role in the pathogenesis of HCC and has become a potential target for the treatment of HCC. Targeted regulation of specific ncRNA expression could reverse sorafenib resistance in HCC.
MiRNAs are stable small RNAs that are expressed abundantly in animals and plants. They can bind to the 3'-untranslated region of the target mRNA, and regulate its expression at the post-transcriptional level. The miRNAs’ abnormal expression and its following abnormal biological regulation are closely related to the occurrence and development of age-related macular degeneration (AMD), including inflammatory response, oxidative stress injury, phagocytosis dysfunction and abnormal angiogenesis. Since the dysregulation of miR-155, miR-125b and miR-34a seems to play a more important role in AMD, these microRNAs may be expected to become the new biomarkers and therapeutic targets for AMD.
Exosomes are a type of tiny vesicles released by cells, which contain bioactive molecules such as proteins, nucleic acids, and lipids secreted by cells. Exosomes released by different cells play an important role in tumor development and metastasis. These exosomes can regulate the tumor microenvironment, promote the tumor growth and invasion, and participate in the process of distant metastasis by carrying specific proteins and nucleic acids. In addition, some biomarkers in exosomes can serve as potential biomarkers for early diagnosis and prognosis evaluation of osteosarcoma. This article reviews the research progress of exosomes in osteosarcoma, aiming to gain a deeper understanding of their mechanisms of action in this disease and provide a reference for the development of new treatment strategies and prognostic evaluation indicators.
Objective To investigate the role of micro RNA-451 (miRNA-451) in promoting the osteogenesis of mesenchymal stem cells (MSCs) by targeting regulatory calcium binding protein 39 (CAB39). Methods pMIR-report and pRL-TK vectors were selected to identify the relationship between miRNA-451 and CAB39 by using dual-luciferase reporter assay. pre-miRNA-451 (group A), anti-miRNA-451 (group C), pre-miRNA negative control (group B), and anti-miRNA negative control (group D) were transfected into the C3H10T1/2 cells, respectively. Then, the cells were collected after osteogenic induction for 7 and 14 days. At 7 and 14 days, the real-time fluorescent quantitative PCR and Western blot assays were performed to detect the related osteogenetic biomarkers [Runx2 and alkaline phosphatase (ALP) mRNA] and expressions of CAB39 protein. At 14 days, the extracellular calcium deposition during the osteogenesis of MSCs was tested by Alizarin red staining method. Results CAB39 was the target gene of miRNA-451. At 7 and 14 days after osteogenic induction, the mRNA expressions of Runx2 and ALP in group A were significantly higher than those in group B (P lt; 0.05), and the expressions in group C was significantly lower than those in group D (P lt; 0.05). Furthermore, at 14 days after osteogenic induction, the protein expression of CAB39 in group A (0.55 ± 0.05) was significantly lower than that in group B (1.00 ± 0.07), and the protein expression in group C (1.21 ± 0.05) was significantly higher than that in group D (1.00 ± 0.04), all showing significant difference (P lt; 0.05). Finally, at 14 days after osteogenic induction, the extracellular calcium deposition in group A was obviously more than that in group B, and group C was downregulated when compared with group D. Conclusion miRNA-451 can promote the osteogenesis process of MSCs by downregulating the CAB39.