<sec> <title>Objective:</title> The purpose of this research is to explore the influences of thymosin β4 (Tβ4) in deepsecond-degree scald wound healing of rat skin and its relationship with Wnt/β-catenin pathway. </sec>
<sec> <title>Methods:</title> Deep second-degree scalded model rats were prepared and divided into normal saline (NS) treatment group, Tβ4 treatment group and FH535 inhibitor group. Then, the concentrations of inflammatory factors in the rats were monitored
via adopting the correlated TNF-α and IL-1β ELISA kits. In the meantime, the wound healing rate was analyzed via photography. Subsequently, the qRTPCR procedure was wielded to determine Wnt1 and β-catenin expression in wound tissues, and the degree of wound
tissue injury was examined via hematoxylin and eosin (HE) staining. Finally, Western blotting (WB) was adopted to assess Wnt/β-catenin pathway-associated protein levels. </sec> <sec> <title>Results:</title> Releasing amount of TNF-α
and IL-1β were conspicuously up-regulated after scalding (p <0.01), and Wnt1 and β-catenin expression at molecular transcription level was also significantly raised (p < 0.01). Besides, treatment with 18 μg of Tβ4 significantly
increased the wound healing rate of scalded rats (p < 0.01). In addition, Tβ4 treatment significantly promoted wound healing (p < 0.01) and increased the Wnt1 and β-catenin expression levels (p < 0.01). Moreover, FH535 significantly restrained
the Wnt/β-catenin pathway-correlated protein levels (p < 0.01) and wound healing. </sec> <sec> <title>Conclusion:</title> Tβ4 can promote scald wound healing in rats and may play a role via evoking Wnt/β-catenin
pathway activation. </sec>
Osteoarthritis (OA) is a chronic and inflammatory disease, leading to pain or even disability in severe cases. LncRNA PCGEM1 (PCGEM1) is reported to be dysregulated, serving as critical regulators in various human diseases, including OA. However, the biological role of PCGEM1 and its
underlying mechanisms during OA remained unclear. In the present study, CHON-001 cells were exposed to interleukin (IL)-1β to construct the OA cell model. Expression of PCGEM1 and miR-152-3p in cells was determined by quantitative real-time polymerase chain reaction (qRT-PCR) assay.
Corresponding commercial kits were used to measure the expressions of lactate dehydrogenase (LDH), inter-leukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Protein levels of apoptosis-related proteins, cleaved-Caspase3 and Caspase3, were detected by Western blotting. 3-(4,
5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) tetrazolium (MTT) and flow cytometry assays were utilized for the determination of cell proliferation and apoptosis. The association between PCGEN1 and miR-152-3p was confirmed by a dual-luciferase reporter assay. From the results,
PCGEM1 expression was significantly increased while miR-152-3p was inhibited in CHON-001 cells after IL-1β treatment. In addition, silencing of PCGEM1 could promote proliferation, inhibit the apoptosis, suppress LDH level and alleviate inflammation response caused by IL-1β
in CHON-001 cells by sponging miR-152-3p. In a word, PCGEM1 down-regulation suppressed OA progression by the regulation of miR-152-3p expression, functioning as a potential therapeutic target for OA clinical treatment.
MicroRNAs (miRNAs/miRs) have been identified to serve a key role in the development of tumors. However, the role of miR-133b in colorectal cancer (CRC) remains largely unclear. This study will investigate the role and mechanism of miR-133b in CRC. Reverse transcription-quantitative
polymerase chain reaction analysis was performed to detect the level of miR-133b in CRC cell lines. Bioinformatics software TargetScan predicted the potential target genes of miR-133b, and a dual luciferase reporter assay was used to confirm this. To investigate the role of miR-133b in CRC
cells, miR-133b was upregulated or downregulated in CRC cell lines (SW620 and HT-29) by transfecting with a miR-133b mimic or inhibitor, respectively. Subsequently, cell viability was analyzed using MTT assay, whereas cell apoptosis and the cell cycle distribution were analyzed by flow cytometry.
In addition, the associated protein levels were detected using western blot analysis. The results demonstrated that miR-133b was significantly downregulated in CRC cell lines when compared with the normal colonic epithelial NCM-460 cell line. Human antigen R (HuR; also termed ELAVL1) was demonstrated
to be a direct target of miR-133b and was negatively regulated by miR-133b. HuR was also notably upregulated in the CRC cell lines when compared with the normal control. Transfection of SW620 and HT-29 cells with the miR-133b mimic significantly inhibited cell viability, and induced cell apoptosis
and G1 phase arrest, while upregulation of HuR demonstrated the opposite effects. Furthermore, the present data demonstrated that the miR-133b mimic significantly enhanced the protein levels of p21 and p27, and downregulated cyclin D1 and cyclin A levels in SW620 and HT-29 cells;
the opposite effects were observed following treatment with the miR-133b inhibitor. In conclusion, the data indicate that miR-133b suppressed CRC cell growth by targeting HuR.
This study evaluated the effect of upgrading the quality of maize stover (MS) on milk nutritive value. The study involved feeding MS improved using urea (U), chopped groundnut stover (cGS), chopped soybean stover (cSS), mineralized groundnut stover solution (mGS) and mineralized soybean stover solution (mSS) to lactating dairy cows. The feeding trial involved twelve (12) dairy cows in their second parity. Effect of supplementation with MS improved with U, cGS, cSS, mGS and mSS on milk quality was evaluated following on-station feeding trials. The study involved 22 factorial experiments within a Completely Randomised Design (CRD). Milk samples were analysed for protein, lactose, fat and solid not fat (SNF). Mean milk protein levels ranged from 3.52mg/ml to 3,73mg/ml (s.e=0.03) for milk from cows fed on MS improved using cGS and mGS respectively. Protein and Lactose were observed to be the least variable (3.64g/ml ±0.12, and 5.24g ±0.24 respectively). Average milk fat content was highest (4.78%, se=0.52) in milk from cows fed on UET treated MS and lowest (3.43%, se=0.52) in milk from cows fed on gGS protein based MS. Within legume type milk fat was higher(4.75%±1.99) in milk from cows fed on MS blended with mGS than that in milk from cows fed on MS improved with cGS (3.43%±1.99). Similar result was observed in milk fat from cows fed on MS improved with the use of soybean. Lactose in milk from cows fed on UET treated MS was highest (5.51g, se=0.061) and lowest (5.10g, se=0.061) in milk from cows fed on MS blended with cGS. Milk from cows fed on MS improved with mGS was higher (9.61p/cwt, se=0.14) in SNF and lowest (8.88p/cwt, se=0.14) in milk from cows fed on MS with cGS. The milk density values ranged from 32.65sg, se=0.53 for milk from cows fed on UET treated MS to 30.42sg, se=0.053 for milk from cows fed on MS blended with cGS. Milk components were higher when cows were fed on MS improved using mineralized legume stover solutions.
AIM: To determine the role of heparanase-1 (HPSE-1) in orbital rhabdomyosarcoma (RMS), and to investigate the feasibility of HPSE-1 targeted therapy for RMS.
METHODS: Immunohistochemistry was performed to analyze HPSE-1 expression in 51 cases of orbital RMS patients (including 28 cases of embryonal RMS and 23 cases of alveolar RMS), among whom there were 27 treated and 24 untreated with preoperative chemoradiotherapy. In vitro, studies were conducted to examine the effect of HPSE-1 silencing on RMS cell proliferation and tube formation of human umbilical vein endothelial cells (HUVECs). RD cells (an RMS cell line) and HUVECs were infected with HPSE-1 shRNA lentivirus at a multiplicity of infection (MOI) of 10 and 30 separately. Real-time PCR and Western blot were applied to detect the mRNA and protein expression levels of HPSE-1. Cell viability of treated or control RD cells was evaluated by cell counting kit-8 (CCK-8) assay. Matrigel tube formation assay was used to evaluate the effect of HPSE-1 RNAi on the tube formation of HUVECs.
RESULTS: Immunohistochemistry showed that the expression rate of HPSE-1 protein was 92.9% in orbital embryonal RMS and 91.3% in orbital alveolar RMS. Tissue from alveolar orbital RMS did not show relatively stronger staining than that from the embryonal orbital RMS. However, despite the types of RMS, comparing the cases treated chemoradiotherapy with those untreated, we have observed that chemoradiotherapy resulted in weaker staining in patients' tissues. The expression levels of HPSE-1 declined significantly in both the mRNA and protein levels in HPSE-1 shRNA transfected RD cells. The CCK-8 assay showed that lentivirus-mediated HPSE-1 silencing resulted in significantly reduced RD cells viability in vitro. Silencing HPSE-1 expression also inhibited VEGF-induced tube formation of HUVECs in Matrigel.
CONCLUSION: HPSE-1 silencing may be a promising therapy for the inhibition of orbital RMS progression.
Atherosclerosis (AS) seriously impairs the health of human beings and is manifested initially as endothelial cells (ECs) impairment and dysfunction in vascular intima, which can be alleviated through mobilization of endothelial progenitor cells (EPCs) induced by stromal-cell-derived factor-1α (SDF-1α). A strong inverse correlation between HDL and AS has been proposed. The aim of the present work is to investigate whether 4F, an apolipoprotein A-I (apoA-I, major component protein of HDL) mimic peptide, can upregulate SDF-1α in mice and human umbilical vein endothelial cells (HUVECs) and the underlying mechanism. The protein levels of SDF-1α were measured by ELISA assay. Protein levels of HIF-1α, phosphorylated Akt (p-Akt), and phosphorylated ERK (p-ERK) were evaluated by Western blotting analysis. The results show that L-4F significantly upregulates protein levels of HIF-1α, Akt, and ERK, which can be inhibited by the PI3K inhibitor, LY294002, or ERK inhibitor, PD98059, respectively. Particularly, LY294002 can downregulate the levels of p-ERK, while PD98059 cannot suppress that of p-Akt. D-4F can upregulate the levels of HIF, p-Akt, and p-ERK in the abdominal aorta and inferior vena cava from mice. These results suggest that 4F promotes SDF-1α expression in ECs through PI3K/Akt/ERK/HIF-1α signaling pathway.
This study was conducted to evaluate the effects of dietary supplementation of different recommended levels of Cu and Zn on antioxidant capacity, tissue mineral status, minerals excretion, meat quality, digestive enzyme activity, and metal transporters in finishing pigs. A total of 120 pigs (with an average initial body weight (BW) of 70.0 ± 2.1 kg) were randomly divided into four treatments: (1) basal diet without added Cu or Zn (control), (2) basal diet+35 mg cupreous N-carbamylglutamate chelate (NCG-Cu) +150 mg zinc-methionine chelate (Zn-Met) (AC), (3) basal diet + 3.0 mg of NCG-Cu + 43 mg Zn-Met (CN), and (4) basal diet + 3.5 mg NCG-Cu + 50 mg Zn-Met (NRC100). Pig growth performance was not affected by the level of Cu or Zn. Among the four treatments, the AC treatment had the highest concentration (P < 0.05) of glutathione peroxidase (GSH-Px). Pigs fed the AC diet had the highest (P < 0.05) liver Zn, fecal Cu, and fecal Zn among the four treatments. The protein levels of trypsin and aminopeptidase N (APN) in the intestinal mucosa showed their highest levels (P < 0.05) in the NRC100 and AC treatments. The mRNA levels of trypsinogen and APN were significantly up-regulated (P < 0.05) in the AC, CN, and NRC100 treatments compared with the control. The mRNA levels for the Zn transporter genes SLC30A1 (ZnT1) and SLC30A2 (ZnT2) were significantly up-regulated (P < 0.05) in the AC treatment, and the mRNA levels for SLC39A4 (ZIP4) and metallothionein 1 (MT) in the AC, CN, and NRC100 treatments were significantly up-regulated (P < 0.05) compared with the control. Meat quality were not affected (P > 0.05) by the different recommended levels of Cu and Zn. These results indicated that the supplemental Cu and Zn levels routinely used in AC diets in Chinese commercial feed enterprises should be reduced.
Regenerating liver phosphatase 1 (PRL1) is an established oncogene in various cancers, although its biological function and the underlying mechanisms in glioblastoma multiforme (GBM) remain unclear. Here, we showed that PRL1 was significantly upregulated in glioma tissues and cell lines, and positively correlated with the tumor grade. Consistently, ectopic expression of PRL1 in glioma cell lines significantly enhanced their tumorigenicity and invasion both in vitro and in vivo by promoting epithelial-mesenchymal transition (EMT). Conversely, knocking down PRL1 blocked EMT in GBM cells, and inhibited their invasion, migration and tumorigenic growth. Additionally, PRL1 also stabilized Snail2 through its deubiquitination by activating USP36, thus revealing Snail2 as a crucial mediator of the oncogenic effects of PRL1 in GBM pathogenesis. Finally, PRL1 protein levels were positively correlated with that of Snail2 and predicted poor outcome of GBMs. Collectively, our data support that PRL1 promotes GBM progression by activating USP36-mediated Snail2 deubiquitination. This novel PRL1/USP36/Snail2 axis may be a promising therapeutic target for glioblastoma.
Aging is important medical and social problem. Excessive angiopoietin-like protein (ANGPTL)-2 signaling causes chronic tissue inflammation, promoting development and progression of aging-related diseases. Moreover, circulating ANGPTL2 levels reportedly predict risk of some aging-related diseases and subsequent death. However, there are as yet no reports of whether circulating ANGPTL2 levels predict vital prognosis in younger-old, community-dwelling populations. This study investigated associations between plasma ANGPTL2 levels and all-cause and specific-cause mortality in this population. The case-cohort study was abstracted from an on-going, age-specific prospective cohort study: the New Integrated Suburban Seniority Investigation Project. This project enrolled 3073 participants aged 64 years at the beginning of the investigation from 1996 through 2005. A sub-cohort of 714 randomly sampled participants plus 387 cases representing deceased participants followed through 2015 underwent survival analysis. Plasma ANGPTL2 concentrations were positively associated with >80% and 100% higher risk of all-cause mortality and cancer mortality, respectively, after adjustment for gender, smoking, alcohol consumption, walking time, sleep duration, caloric intake, medical status, disease history, BMI, and triglyceride, creatinine, uric acid, and high sensitivity C-reactive protein levels. More robust association between ANGPTL2 levels and all-cause and cancer mortality was seen in subjects with either frailties or with lifestyles of heavier drinking or current smoking. Elevated plasma ANGPTL2 levels are associated with high all-cause and cancer mortality in a community-dwelling sample of younger-old adults. These findings expand our knowledge of human aging and associated diseases.