Catalpol attenuates ischemic stroke by promoting neurogenesis and angiogenesis via the SDF-1α/CXCR4 pathway.

BACKGROUND: Stroke is a leading cause of disability and death worldwide. Currently, there is a lack of clinically effective treatments for the brain damage following ischemic stroke. Catalpol is a bioactive compound derived from the traditional Chinese medicine Rehmannia glutinosa and shown to be protective in various neurological diseases. However, the potential roles of catalpol against ischemic stroke are still not completely clear. PURPOSE: This study aimed to further elucidate the protective effects of catalpol against ischemic stroke. METHODS: A rat permanent middle cerebral artery occlusion (pMCAO) and oxygen-glucose deprivation (OGD) model was established to assess the effect of catalpol in vivo and in vitro, respectively. Behavioral tests were used to examine the effects of catalpol on neurological function of ischemic rats. Immunostaining was performed to evaluate the proliferation, migration and differentiation of neural stem cells (NSCs) as well as the angiogenesis in each group. The protein level of related molecules was detected by western-blot. The effects of catalpol on cultured NSCs as well as brain microvascular endothelial cells (BMECs) subjected to OGD in vitro were also examined by similar methods. RESULTS: Catalpol attenuated the neurological deficits and improved neurological function of ischemic rats. It stimulated the proliferation of NSCs in the subventricular zone (SVZ), promoted their migration to the ischemic cortex and differentiation into neurons or glial cells. At the same time, catalpol increased the cerebral vessels density and the number of proliferating cerebrovascular endothelial cells in the infracted cortex of ischemic rats. The level of SDF-1α and CXCR4 in the ischemic cortex was found to be enhanced by catalpol treatment. Catalpol was also shown to promote the proliferation and migration of cultured NSCs as well as the proliferation of BMECs subjected to OGD insult in vitro. Interestingly, the impact of catalpol on cultured cells was inhibited by CXCR4 inhibitor AMD3100. Moreover, the culture medium of BMECs containing catalpol promoted the proliferation of NSCs, which was also suppressed by AMD3100. CONCLUSION: Our data demonstrate that catalpol exerts neuroprotective effects by promoting neurogenesis and angiogenesis via the SDF-1α/CXCR4 pathway, suggesting the therapeutic potential of catalpol in treating cerebral ischemia.

Long-term survival in a patient with multiple metastatic gastric cancer treated with PTX plus emvolimab and disitamab vedotin: case report and treatment experience: A case report.

RATIONALE: Most Chinese patients with locally advanced gastric cancer at diagnosis have an overall 5-year survival rate of <50%. Surgical resection alone is not suitable for patients with locally advanced gastric cancer. Currently, comprehensive treatment is the focus of locally advanced gastric cancer. PATIENTS CONCERNS: The patient, a 56-year-old female, was admitted to the hospital because of "4 + months of double hydronephrosis found during a physical examination." Who was admitted for computer tomography and gastroscopy examinations, and take pathological tissue specimens during endoscopic examination. DIAGNOSES: Computed tomography assessment indicated ulcerative gastric cancer with an abdominal implant, bladder, and bone metastases. An endoscopic examination revealed that the ulcer of the gastric angle was huge, and through relevant auxiliary examinations, the diagnosis of this disease is gastric cancer complicated with multiple metastases to bladder, rectum, lumbar spine, and peritoneum. Clinically diagnosed as cT4bN3M1. INTERVENTIONS: The patient is currently undergoing first, second, and third line neoadjuvant therapy, combined with immunotherapy, targeted therapy, neoadjuvant intraperitoneal systemic chemotherapy, nutritional support, and other treatment plans. OUTCOMES: After 15 cycles of treatment, the progression-free survival had reached 15 months. The patient had an NRS2002 score of 1, an ECOG score of I, a quality of life score of 55, albumin of 35.27 g/L, and a decrease in abdominal and pelvic fluid accumulation and exudation compared to before. LESSONS: We demonstrated high survival of almost 3 years in a patient with gastric cancer that was complicated by bone, peritoneal, rectal, and bladder metastases. The combination of immunotherapy, targeted therapy, and neoadjuvant intraperitoneal systemic chemotherapy, along with the maintenance of nutritional status and CTCs could be a valuable modality for the subsequent treatment and observation of similar patients.

Oxidative removal of fluorescent components from soil DOM and its effect on heavy metals around abandoned mining areas.

The oxidation stability of soil organic matter (SOM) plays an important role in the environmental chemical behavior of heavy metals (HMs). In this study, the oxidation stability of SOM and soil dissolved organic matter (DOM) for four soils around the mining area in Western China, including grassland (GR), forest land (FR), farmland soil (FA), and mining area soil (MA), was investigated. The oxidation effect of fluorescent DOM (FDOM) was determined by using synchronous fluorescence spectroscopy (SFS). The results showed that the oxidation stability of SOM for four soils follows the order: MA > GR > FR > FA. Protein-like fluorescence (A2) is dominant in soil DOM, more than 96% of which were more easily degraded. As the wavelength increases, FDOM components become more difficult to oxidize. Second derivative, two-dimensional correlation spectroscopy (2D-COS) and 1/n power transformation can identify more FDOM components, protein-like materials can be preferential removal by the oxidation process, followed by humic-like substances. The oxidation process increased the release of Cr, Cu, Zn, Pb and Fe in FA soil. Therefore, the oxidation stability of SOM and FDOM can affect the immobilization and release of HMs, and this work provides scientific guidance for remediation of soil HMs around abandoned mining areas.

Catalpol Alleviates Ischemic Stroke Through Promoting Angiogenesis and Facilitating Proliferation and Differentiation of Neural Stem Cells via the VEGF-A/KDR Pathway.

Stroke is one of the leading causes of disability and death globally with a lack of effective therapeutic strategies. Catalpol is a bioactive compound derived from the traditional Chinese medicine Rehmannia glutinosa and it has been shown to be protective against various neurological diseases. The potential roles of catalpol against ischemic stroke are still not completely clear. In this study, we examined the effect and mechanism of catalpol against ischemic stroke using in vivo rat distal middle cerebral artery occlusion (dMCAO) and in vitro oxygen-glucose deprivation (OGD) models. We demonstrated that catalpol indeed attenuated the neurological deficits caused by dMCAO and improved neurological function. Catalpol remarkably promoted angiogenesis, promoted proliferation and differentiation of neural stem cells (NSCs) in the subventricular zone (SVZ), and prevented neuronal loss and astrocyte activation in the ischemic cortex or hippocampal dentate gyrus (DG) in vivo. The vascular endothelial growth factor receptor 2 (KDR, VEGFR-2) inhibitor SU5416 and VEGF-A shRNA were used to investigate the underlying mechanisms. The results showed that SU5416 administration or VEGF-A-shRNA transfection both attenuated the effects of catalpol. We also found that catalpol promoted the proliferation of cultured brain microvascular endothelial cells (BMECs) and the proliferation and differentiation of NSCs subjected to OGD insult in vitro. Interestingly, the impact of catalpol on cultured cells was also inhibited by SU5416. Moreover, catalpol was shown to protect NSCs against OGD indirectly by promoting BMEC proliferation in the co-cultured system. Taken together, catalpol showed therapeutic potential in cerebral ischemia by promoting angiogenesis and NSC proliferation and differentiation. The protective effects of catalpol were mediated through VEGF-A/KDR pathway activation.

Influence of thermal air oxidation on the chemical composition and uranium binding property of intrinsic dissolved organic matter from biochar.

In this work, uranium (U(VI)) binding characteristics of the intrinsic dissolved organic matters (DOM) from the biochars prepared under thermal air oxidation (TAO) and non-TAO conditions were studied using synchronous fluorescence spectra (SFS) and Fourier transform infrared (FTIR) in conjunction with the general two-dimensional correlation spectroscopy (2D-COS), heterospectral 2D-COS and moving-window (MW) 2D-COS. The chemical compositions of the intrinsic DOMs from biochars with/without TAO were investigated by Fourier transform-ion cyclotron resonance mass spectrometry (FT-ICR MS). Results showed that the preferential binding of U(VI) to functional groups followed the order: 937 (carboxyl γC-OH), 981 (carboxyl γC-OH), 1511 (aromatic vC = C), 1108 (esters or ethers vC-O), 1282 (esters or carboxyl vC-O), 1698 (saturated carboxylic acid or ketone vC = O) cm-1 for biochar DOM after TAO (OB600), and 937 (carboxyl γC-OH), 1484 (lipids δC-H or phenolic vC-O), 1201 (esters or carboxyl vC-O), 1112 (esters or ethers vC-O), 1706 (saturated aldehyde, carboxylic acid or ketone vC = O), 1060 (phenolic, esters or ethers vC-O), 1014 (phenolic, esters or ethers vC-O) cm-1 for the pristine biochar (B600). Fulvic-like substances at 375 nm in the biochar DOM showed a preferential binding with U(VI) after TAO, while humic-like substances played a more critical role in the U(VI) complexation with biochar DOM obtained from non-TAO condition. The results also indicated that TAO increased the content of fluorescent DOM and the chemical stability of DOM-U(VI) complexes. The FT-ICR MS results showed an increase in the relative abundance of protein-like, carbohydrates-like, tannins-like, unsaturated hydrocarbons, and condensed aromatic structure and a decrease in the relative abundance of lipid-like and lignin-like after TAO. Consequently, although biochar after TAO had a much poorer content of intrinsic DOM, its intrinsic DOM showed a much higher capacity in U(VI) precipitation. Therefore, the TAO substantially changed the chemical composition, binding property and environmental behavior of intrinsic DOM from biochar.

Insight into the effects of biological treatment on the binding properties of copper onto dissolved organic matter derived from coking wastewater.

Biological treatment can remove more than 89.8% of total organic carbon (TOC) and 94.4% of fluorescent dissolved organic matter (DOM) in the coking wastewater, thereby affecting the migration, transformation and bioavailability and binding characteristics of heavy metals (HMs). The results of parallel factor analysis (PARAFAC) show that protein-like materials accounted for 97.53% in the coking wastewater DOM, a large number of humic-like substances are produced and accounted for more than 55.40% after biological treatment. A new spectral data processing method, the 1/n-th power transformation after two-dimensional correlated spectroscopy (2D-COS) in combination with synchronous fluorescence spectra (SFS), can identify small features obscured by strong peaks, and reveal more binding sites as well as preserve the sequential order information. The result indicates that the preferential bonding of Cu(II) is at 306 nm (protein-like) for coking wastewater DOM, and at 514 nm (humic-like) for effluent DOM. The C-O group of esters and alcohols can preferentially complexate with Cu(II) in the coking wastewater and effluent DOM. The log KM values of PARAFAC components with Cu(II) are in the range of 3.59-5.06 for coking wastewater DOM, and in the range of 4.80-5.64 for the effluent DOM. Log KM values for protein-like materials with Cu(II) are higher than these for fulvic- and humic-like substances. Humic-like substances can form more stable complexes with Cu(II) in the effluent DOM. Biological treatment increases the chemical stability of DOM-Cu(II) complexes, thereby further reducing the environmental risk of Cu(II).

Cytokine autoantibodies in SARS-CoV-2 prepandemic and intrapandemic samples from an SLE cohort.

Cytokine autoantibodies, particularly those directed to type I interferon (T1IFN), have been reported to portend an increased risk of severe COVID-19. Since SLE is one of the conditions historically associated with T1IFN autoantibodies, we sought to determine the prevalence of cytokine autoantibodies in our local cohort of 173 patients with SLE prepandemic and intrapandemic, of which nine had confirmed exposure to SARS-CoV-2. Autoantibodies to 16 different cytokines, including T1IFN, were measured by an addressable laser bead immunoassay. None of the 9 patients with confirmed exposure to SARS-CoV-2 had autoantibodies to T1IFN and none had severe COVID-19 symptoms, necessitating hospitalisation. Hence, we could not confirm that TIIFN autoantibodies increase the risk for severe COVID-19. In addition, the cytokine autoantibody pattern did not differ between those with and without evidence of SARS-CoV-2 exposure.

Effects of dimethyl sulfoxide pretreatment on the bonding properties of fluorotic dentin of different severity: An in vitro study.

STATEMENT OF PROBLEM: Bonding to fluorotic dentin is weaker than to sound dentin, but methods to improve bonding have not been well addressed. PURPOSE: The purpose of this in vitro study was to investigate the effects of dimethyl sulfoxide (DMSO) pretreatment on the bond strength and resin-dentin surface of fluorotic dentin of different severity. MATERIAL AND METHODS: Phosphoric acid-etched dentin specimens exhibiting mild fluorosis (ML-F), moderate fluorosis (MD-F), and severe fluorosis (SE-F) were randomly bonded with Single Bond 2 (SB2) pretreated with 50% DMSO (experimental groups) or deionized water (control groups). The bonded teeth were sectioned for microshear bond strength (µSBS) testing immediately or after aging, for micromorphology observation of the bonding interface under a scanning electron microscope, and for resin tags and microleakage evaluation under a confocal laser scanning microscope. The degree of conversion of the adhesive resin was calculated by Fourier transform infrared spectroscopy. According to varying bonding steps, the mineralized dentin powders of ML-F, MD-F, and SE-F were randomly divided into 4 subgroups (blank, PA, PA+SB2, and PA+DMSO+SB2) and incubated in artificial saliva to examine the level of enzymatic degradation product of type I collagen. Data were analyzed by using ANOVA and the Tukey test (α=.05). RESULTS: Dental fluorosis and thermocycling had negative effects on µSBS (P<.001), while DMSO pretreatment preserved or even improved µSBS (P<.001). DMSO had no influence on the degree of conversion (P=.618). Significant effects were found for bonding steps (P<.001), but not that of dental fluorosis (P=.131) on the enzymatic degradation product of type I collagen. Images showed sparser and more expanded collagen fibril meshwork, deeper resin penetration, and less microleakage in the experimental groups. CONCLUSIONS: DMSO pretreatment provided increased and durable dentin bonding to fluorotic dentin probably by dispersing collagen fibrils into a sparser network and inhibiting the degradation of type I collagen.

Feasibility of Catalpol Intranasal Administration and Its Protective Effect on Acute Cerebral Ischemia in Rats via Anti-Oxidative and Anti-Apoptotic Mechanisms.

PURPOSE: Catalpol is the main active component of Rehmannia glutinosa, which has a variety of pharmacological activities, including anti-inflammatory and anti-oxidative effects. This study investigates the feasibility of catalpol intranasal administration and its protective effect on acute cerebral ischemia in rats via anti-oxidative and anti-apoptotic mechanisms. PATIENTS AND METHODS: This study investigates the method of catalpol intranasal administration to evaluate the nasal mucosal toxicity, brain targeting and pharmacokinetics of catalpol. The protective effect of catalpol of intranasal administration on stroke-induced brain injury in rats and its mechanisms on oxidative stress pathway Nrf2/HO-1 and apoptosis were also investigated using middle cerebral artery occlusion (MCAO). RESULTS: The results showed that catalpol intranasal administration was safe and feasible with no hemolysis, no bad effect on the maxillary ciliary movement of bullfrog. After intranasal administration, the brain targeting index (DTI) of catalpol was greater than 1, which indicated that catalpol had good brain targeting after intranasal administration. The bioavailability of catalpol administered intranasally was higher than that of in plasma. In MACO model, catalpol intranasal administration could significantly reduce cerebral infarction volume, neurological dysfunction and brain edema. In addition, catalpol intranasal administration can also reduce the brain cell's occurrence of apoptosis, promote the expression of Bcl-2 protein and inhibit the expression of Bax protein, reduce oxidative stress damage via up-regulating expression of Nrf2 and HO-1, increasing the activities of SOD and decreasing the activities of MDA. CONCLUSION: Collectively, catalpol intranasal administration has good safety, stability and brain targeting. It can effectively protect the brain injury of the rat model of acute cerebral ischemia and provide the possibility of drug administration in the acute stage of cerebral ischemia, especially before entering the hospital.

Effect of 45S5 bioactive glass on the microshear bond strength of dental fluorosis.

The aim of this in vitro study was to estimate the effect of the species concentration of 45S5 bioactive glass (BAG) used as pretreatment on the microshear bond strength (MSBS) of dental fluorosis (DF). Based on the Thylstrup and Fejerskov index, 80 teeth were randomly divided equally into four groups: TFI 0, sound dentin; TFI 1-3, mild fluorosis; TFI 4-5, moderate fluorosis; and TFI 6-9, severe fluorosis. Each group was randomized into five subgroups. After preparing the dentin hypersensitivity model of DF, the dentin was pretreated as follows, Subgroup 1: deionized water (Control group); Subgroup 2: 1% BAG; Subgroup 3: 5% BAG; Subgroup 4: 10% BAG, and Subgroup 5: 20% BAG. Stochastically one specimen was selected from each subgroup for scanning electron microscope and energy dispersive spectrometer analysis. After being made of resin-tooth bonding samples, the remains were in water bath at 37 °C for 24 hr. Subsequently, samples from each subgroup were randomly selected to test MSBS without aging, or after a thermocycle of 5,000 and 10,000 times, respectively. The fracture modes were analyzed. Compared with the group of 1% BAG and Control, the exposure area of tubules in 5%, 10%, and 20% BAG group had significant difference (p < .05). MSBS results indicated that there were significant differences between 10% BAG with other groups. The 20% BAG group showed the lowest MSBS among all groups. Pretreatment of 10% BAG solution may be conductive to enhance the bond strength of DF, while 20% BAG solution adversely.

Targeting the Nod-like receptor protein 3 Inflammasome with inhibitor MCC950 rescues lipopolysaccharide-induced inhibition of osteogenesis in Human periodontal ligament cells.

OBJECTIVE: We aim to investigate whether lipopolysaccharide-stimulated activition of Nod-like receptor protein 3 (NLRP3) Inflammasome inhibits osteogenesis in Human periodontal ligament cells (HPDLCs). Futhermore, to study whether MCC950 (a inhibitor of NLRP3 Inflammasome) rescues lipopolysaccharide-induced inhibition of osteogenesis in HPDLCs as well as the underlying mechanisms. METHODS: HPDLCs were isolated from periodontal ligament of healthy orthodontic teeth from teenagers, and cells surface marker protein were detected by flow cytometry. Cells viability were determined by Cell Counting kit 8 assay. Enzyme-linked immunosorbent assay was used to analyze the secretion of proinflammatory factors. Western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were measured assessing the expression of NLRP3 and Caspase-1. RT-qPCR, Alizarin red staining and Alkaline phosphatase staining were tested to determine the osteogenic differentiation capacity of HPDLCs. RESULTS: It was found that lipopolysaccharide in the range of concentrations from 10 to 100 µg/ml significantly inhibited HPDLCs viability at 24 h and significantly improved proinflammatory cytokine expressions at 8 h and 24 h. MCC950 reversed lipopolysaccharide-stimulated proinflammatory cytokine expressions including interleukin-1ß and interleukin-18, but not tumor necrosis factor-α. In addition, MCC950 rescued the lipopolysaccharide-inhibited osteogenic gene (Alkaline phosphatase, Runt-related transcription factor 2, and Osteocalcin). Moreover, MCC950 downregulated lipopolysaccharide-induced relative protein of NLRP3 Inflammasome signaling pathway, such as NLRP3 and Caspase-1. CONCLUSION: MCC950 rescues lipopolysaccharide-induced inhibition of osteogenesis in HPDLCs via blocking NLRP3 Inflammasome signaling pathway, and it may be used as a promising therapeutic agent for periodontitis or periondontal regenerative related disease.

Micro-RNA-451 Reduces Proliferation of B-CPAP Human Papillary Thyroid Cancer Cells by Downregulating Expression of Activating Transcription Factor 2.

BACKGROUND MicroRNAs (miRNAs) are novel biomarkers that are important in tumorigenesis and cancer treatment resistance. miR-451 is expressed in human papillary thyroid carcinoma (PTC) tissues and is associated with tumor progression. This study investigated the molecular mechanism associated with the effects of miR-451 on B-CPAP human PTC cells in vitro. MATERIAL AND METHODS Binding of miRNAs to the 3' untranslated region (3'UTR) of messenger RNA (mRNA) was determined with a luciferase reporter assay. miRNAs and plasmids were transfected into human PTC B-CPAP cells with Lipofectamine 2000 Transfection Reagent. Cell viability was tested with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay. The levels of miRNAs and mRNA were determined with quantitative polymerase chain reaction and protein levels were analyzed with immunoblotting. RESULTS miR-451 bound to wild-type but not mutant 3'-UTR of activating transcription factor 2 (ATF2). MiR-451 mimics inhibited the growth of B-CPAP cells and reduced mRNA and protein levels in ATF2, whereas miR-451 inhibitors promoted the growth of B-CPAP cells and increased mRNA and protein levels in ATF2. CONCLUSIONS miR-451 directly bound to the 3'UTR of ATF2, decreased mRNA and protein levels in ATF2, and inhibited growth of B-CPAP cells. Our findings suggest that miR-451 may be a potential therapeutic target for PTC.

Network Pharmacology-Based Prediction of Catalpol and Mechanisms against Stroke.

AIM: To apply the network pharmacology method to screen the target of catalpol prevention and treatment of stroke, and explore the pharmacological mechanism of Catalpol prevention and treatment of stroke. METHODS: PharmMapper, GeneCards, DAVID, and other databases were used to find key targets. We selected hub protein and catalpol which were screened for molecular docking verification. Based on the results of molecular docking, the ITC was used to determine the binding coefficient between the highest scoring protein and catalpol. The GEO database and ROC curve were used to evaluate the correlation between key targets. RESULTS: 27 key targets were obtained by mapping the predicted catalpol-related targets to the disease. Hub genes (ALB, CASP3, MAPK1 (14), MMP9, ACE, KDR, etc.) were obtained in the key target PPI network. The results of KEGG enrichment analysis showed that its signal pathway was involved in angiogenic remodeling such as VEGF, neurotrophic factors, and inflammation. The results of molecular docking showed that ACE had the highest docking score. Therefore, the ITC was used for the titration of ACE and catalpol. The results showed that catalpol had a strong binding force with ACE. CONCLUSION: Network pharmacology combined with molecular docking predicts key genes, proteins, and signaling pathways for catalpol in treating stroke. The strong binding force between catalpol and ACE was obtained by using ITC, and the results of molecular docking were verified to lay the foundation for further research on the effect of catalpol on ACE. ROC results showed that the AUC values of the key targets are all >0.5. This article uses network pharmacology to provide a reference for a more in-depth study of catalpol's mechanism and experimental design.

The effects of 45S5 bioactive glass and Er:YAG Laser on the microtensile bond strength of fluorosed teeth.

This vitro study aimed to evaluate the effects of 45S5 bioactive glass (BAG) and Er:YAG laser as desensitization treatments on the microtensile bond strength (MTBS) of fluorosed teeth. The 120 noncarious fluorosis were to obtain superficial dentin, being classified into four groups according to the Thylstrup and Fejerskov Index (TFI). Specimens from each group were randomly divided into five subgroups. After fluorosed teeth hypersensitivity models were established, the following pretreatments were applied on dentine surface: Subgroup 1: deionized water (Control); Subgroup 2: BAG; Subgroup 3: Er:YAG laser; Subgroup 4: BAG + Er:YAG laser, and Subgroup 5: Er:YAG laser + BAG. One sample was randomly selected from each subgroup for scanning electron microscope (SEM). The remaining samples were bonded with composite resin by Adper Single Bond 2 adhesive. Then water bath at 37°C for 24 hr. After 5,000 thermocycling, MTBS was tested and fracture mode was analyzed. The difference of MTBS between BAG group and Control group was found statistically significant (p < .05) in fluorosis. The Er:YAG laser + BAG group showed lowest MTBS values in fluorosis. In conclusion, the pretreatment of BAG might be beneficial to the adhesive of fluorosed teeth. Er:YAG laser desensitization alone or using BAG first and then Er:YAG laser desensitization might not affect the adhesive of fluorosed teeth, while Er:YAG laser desensitization followed by the pretreatment of BAG would be not conducive to the adhesive of fluorosed teeth.

TLE2 is associated with favorable prognosis and regulates cell growth and gemcitabine sensitivity in pancreatic cancer.

BACKGROUND: The transducin-like enhancer of split (TLE) proteins are a group of transcriptional corepressors. They play a crucial role in cellular homeostasis and are involved in various cancers. Compared with other TLE family members, little is known about the role and the underlying mechanism of TLE2 in human cancers. This study aimed to investigate the role of TLE2 in pancreatic ductal adenocarcinoma (PDAC) using in silico analysis and in vitro experiments. METHODS: Data were obtained from the Cancer Genome Atlas (TCGA) database to evaluate the prognostic value of TLE2 in PDAC. The MiaPaCa-2 cell line was transfected with siRNA to inhibit endogenous TLE2 expression, and a PANC-1 cell line with stable TLE2 overexpression was constructed using lentiviral transfection, which were confirmed by real-time polymerase chain reaction and western blotting. MTT assay, transwell invasion assays, and flow cytometry were carried out to assess cell viability, invasion, and apoptosis, respectively. TLE2 expression in PDAC cells was altered to evaluate their sensitivity to gemcitabine. Gene set enrichment analysis (GSEA) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were conducted to predict the biological role of TLE2. RESULTS: High expression of TLEs was significantly associated with increased overall survival (OS) and disease-free survival (DFS) in patients with PDAC. Among the PDAC cell lines, TLE2 expression was lowest and highest in PANC-1 cells and MiaPaCa-2 cells, respectively. TLE2 overexpression impaired the proliferation ability of PANC-1 cells and downregulation of TLE2 promoted the proliferation of MiaPaCa-2 cells. Upregulation of TLE2 in PANC-1 cells induced S-phase accumulation and sensitivity to gemcitabine. In contrast, the downregulation of TLE2 in MiaPaCa-2 cells promoted resistance to gemcitabine. Moreover, bioinformatics analysis also revealed the potential tumor suppressor role of TLE2 and uncovered a close relationship between TLE2 expression and cell cycle regulation. CONCLUSIONS: Our results suggest that TLE2 expression is correlated with prognosis in patients with PDAC and show that TLE2 plays a central role in the regulation of cell proliferation, the cell cycle, and gemcitabine sensitivity. This study provides new insights and evidence that TLE2 functions as a tumor suppressor gene and prognostic marker in PDAC.

A Monoclonal Antibody to M-Phase Phosphoprotein 1/Kinesin-Like Protein KIF20B.

Kinesin-like protein KIF20B, originally named M-phase phosphoprotein 1 (MPP1), is a plus-end-directed kinesin-related protein that exhibits in vitro microtubule-binding and -bundling properties as well as microtubule-stimulated ATPase activity. It has been characterized as a slow molecular motor that moves toward the plus-end of microtubules. Human autoantibodies directed against KIF20B have been described in up to 25% of patients with idiopathic ataxia and less commonly in other neuropathies and autoinflammatory conditions. One of the limitations of research into the structure and function of KIF20B has been a reliable monoclonal antibody that can be used in a variety of applications. To establish a reference standard for anti-KIF20B immunoassays and facilitate studies on the role of KIF20B in developmental cell biology, we developed an IgG1 monoclonal antibody, 10C7, which reacts with the cognate KIF20B protein in Western immunoblots and in addressable laser bead immunoassays. In HEp2 cells, leptomeningeal pericytes, and transfected HEK293T cells, indirect immunofluorescence studies showed that reactivity was mainly localized to a proportion of interphase nuclei, but during metaphase, it was redistributed throughout the cytoplasm and perichromatin mass. Later in telophase/anaphase, KIF20B was localized to the stem body and midzone of the midbody. 10C7 also showed remarkable staining of a subset of cells in the cerebellum, ovary, and testis tissues. KIF20B was shown to have extensive coiled-coil domains. The monoclonal antibody, 10C7, will be of value to diagnostic laboratory scientists interested in having a reliable reference standard for anti-KIF20B immunoassays as well as cell, molecular, and developmental biology researchers.

Mechanical, barrier and antimicrobial properties of corn distarch phosphate/nanocrystalline cellulose films incorporated with Nisin and ε-polylysine.

Antimicrobial bio-nanocomposite films were prepared by incorporating nisin (0.25-0.5% W/W) and ε-polylysine (PL, 0.2% W/W) into corn distarch phosphate/nanocrystalline cellulose based films (CN) via casting method. Nisin and PL had significant effects on color parameters (L*, a*, b* and ∆E*) and improved the mechanical, barrier properties, thermal stability of the films. The CN bio-nanocomposite films incorporation the combination of nisin and PL had synergistic antimicrobial activity against Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). Structural property assessment by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) revealed that a clear interaction between the hydroxyl groups of corn distarch phosphate and the amino groups of nisin and PL, leading to the microstructure of the CN bio-nanocomposite films with appropriate content of nisin and PL was more continuous and compact. These results indicate that the CN bio-nanocomposite films containing nisin and PL is a potential active packaging material with enhanced physicochemical properties in food industry.

Scurfy Mice Develop Features of Connective Tissue Disease Overlap Syndrome and Mixed Connective Tissue Disease in the Absence of Regulatory T Cells.

Due to a missense mutation in the Foxp3 gene, scurfy mice are deficient in functional regulatory T cells (Treg). The consequent loss of peripheral tolerance manifests itself by fatal autoimmune mediated multi-organ disease. Previous studies have outlined the systemic inflammatory disease and demonstrated production of anti-nuclear antibodies (ANA) in scurfy mice. However, specific autoantibody targets remained to be defined. ANA are immunological markers for several connective tissue diseases (CTD) and target a large number of intracellular molecules. Therefore, we examined scurfy sera for the presence of different ANA specificities and further assessed the organ involvement in these animals. Indirect immunofluorescence was used as a screen for ANA in the sera of scurfy mice and dilutions of 1/100 were considered positive. Addressable laser bead immunoassays (ALBIA) were used to detect specific autoantibody targets. Subsequent histological tissue evaluation was verified by hematoxylin and eosin (H&E) staining. In our study, we observed that nearly all scurfy mice produced ANA. The most prevalent pattern in scurfy sera was nuclear coarse speckled, also known as the AC-5 pattern according to the International Consensus on ANA Patterns. U1-ribonucleoprotein (U1RNP) was found to be the most common target antigen recognized by autoantibodies in scurfy mice. Additionally, scurfy mice exhibited a mild myositis with histological characteristics similar to polymyositis/dermatomyositis. Myopathy-specific autoantibody profile revealed significantly increased levels of anti-SMN (survival of motor neuron) as well as anti-Gemin3 antibodies in scurfy sera. Overall, we demonstrate that the impaired peripheral tolerance in the absence of regulatory T cells in scurfy mice is associated with features of mixed connective tissue disease (MCTD). This includes, along with our previous findings, very high titers of anti-U1RNP antibodies and an inflammatory myopathy.

MicroRNA-451 as a prognostic marker for diagnosis and lymph node metastasis of papillary thyroid carcinoma.

BACKGROUND AND OBJECTIVE: To investigate the diagnostic potentials of microRNA-451(miR-451) in papillary thyroid carcinoma (PTC) diagnosis and lymph node (LN) metastasis, formalin-fixed, paraffin-embedded (FFPE) tissue specimens corresponding to PTC tumors (n= 60) and their normal counterparts (Normal tissues Adjacent to Tumor, NAT, n= 60), along with sera from PTC patients with malignant tumors (n= 70) and benign lesions (n= 70) were analyzed for the expression of miR-451 by real-time PCR. METHODS: The usefulness of miR-451 expression as a prognostic marker for diagnosis of PTC malignancies was evaluated by Receiver Operating Curve (ROC). We reported that when compared to those in NAT, the levels of miR-451 in FFPE tissues from various stages of PTC patients (n= 60) were significantly lower (Mean ± SEM; 12.62 ± 1.73 vs 38.8 ± 3.51, p< 0.0001). Receiver operating curve (ROC) analysis revealed that the area under curve (AUC) was 0.808; suggesting miR-451 expression was a reliable tissue biomarker for PTC malignancies. Further in depth analyses of these specimens revealed that miR-451 levels were significantly lower in PTC patients with lymph node (LN) metastasis than those without LN metastasis (3.96 ± 1.67 vs. 14.15 ± 1.95, p= 0.006) with calculated AUC of 0.792, supporting the notion that miR-451 expression was also a good indicator for PTC lymph node involvements. Analyses sera from the cohorts of PTC patients indicated that miR-451 levels in patients with malignant lesions (n= 70) were significantly lower (10.72 ± 1.52 vs. 19.28 ± 2.73, p= 0.010) than those with benign ones (n= 70). Parallel analyses of serum miR-451 levels in patients with LN metastasis also showed that they were significantly lower when compared to those without LN metastasis (6.79 ± 2.29 vs. 12.08 ± 1.86, p= 0.017). RESULTS: ROC analyses revealed that AUC was 0.626 for malignancies and was 0.690 for lymph node involvement, respectively, suggesting that miR-451was a modest blood based biomarker for PTC malignancies and lymph node metastasis. CONCLUSIONS: We concluded that miR-451 expression is a reliable FFPE tissue biomarker for PTC malignancies and it may have potentials to become a noninvasive, blood-based biomarker for PTC diagnosis and evaluation of LN status.

Coactivator-associated arginine methyltransferase 1 promotes cell growth and is targeted by microRNA-195-5p in human colorectal cancer.

The pathogenesis of colorectal cancer remains poorly understood. Here, we show that coactivator-associated arginine methyltransferase 1 is frequently upregulated in colorectal cancer tissues and promotes cell growth in vitro and in vivo. Using bioinformatics-based prediction and luciferase reporter system, we found that coactivator-associated arginine methyltransferase 1 is post-transcriptionally targeted by microRNA-195-5p in colorectal cancer. Ectopic expression of microRNA-195-5p led to the suppression of the coactivator-associated arginine methyltransferase 1 3'-untranslated regions activity and downregulation of the endogenous coactivator-associated arginine methyltransferase 1 protein in colorectal cancer cells. Expression analysis verified that microRNA-195-5p was markedly downregulated in human colorectal cancer tissues, which was negatively correlated with the elevated levels of coactivator-associated arginine methyltransferase 1 protein. Enhanced levels of microRNA-195-5p in colorectal cancer cells resulted in a sharp reduction of cell proliferative and colony-formative capacities in vitro. Remarkably, restoration of coactivator-associated arginine methyltransferase 1 in microRNA-195-5p-transfected colorectal cancer cells partially abrogated the inhibition of cell proliferation and colony formation mediated through microRNA-195-5p. These data confirm that microRNA-195-5p might function as an anti-tumor microRNA in colorectal cancer exerting critical control over coactivator-associated arginine methyltransferase 1 expression. The newly identified microRNA-195-5p/coactivator-associated arginine methyltransferase 1 axis may act as a novel promising therapeutic target for colorectal cancer treatment.