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Objective:To investigate the effect of miRNA-3653-3p (miR-3653-3p) on the proliferation and invasion ability of endometrial cancer cells and its related mechanisms.Methods:The data of 356 endometrial cancer patients were downloaded from the OncoLnc database (http://www.oncolnc.org, updated version 2020), and the Kaplan-Meier method was used to analyze the relationship between the expression level of miR-3653-3p and the overall survival of endometrial cancer patients. The miRGator database (https://bio.tools/mirgator_v2.0, updated version 2019) was used to predict the target gene binding to miR-3653-3p. Human endometrial cancer cell lines AN3CA, HEC-1A, HEC-1B, Ishikawa and human normal endometrial epithelial cell line ESC were selected, and the relative expression level of miR-3653-3p was detected by using quantitative real-time fluorescent polymerase chain reaction (qRT-PCR). The cell line with the lowest expression of miR-3653-3p was selected as the research object, which was divided into the negative control group and miR-3653-3p group, and transfected with the control empty vector plasmid and miR-3653-3p overexpression plasmid. CCK-8 method was used to detect the proliferation ability of cells, Transwell method was used to detect the invasion ability of cells, and qRT-PCR and Western blot were used to detect the expression of miR-3653-3p target gene. The effect of miR-3653-3p on the related protein expression of Wnt- β-catenin signaling pathway was detected by using Western blot.Results:Data analysis in the OncoLnc database showed that compared with endometrial cancer patients with low miR-3653-3p expression, patients with high miR-3653-3p expression had better overall survival ( P < 0.01). Compared with human normal endometrial epithelial ESC, the expression levels of miR-3653-3p in endometrial cancer cell lines AN3CA, HEC-1A, HEC-1B, and Ishikawa were all decreased (all P < 0.05), and the relative expression level of miR-3653-3p was the lowest in HEC-1A cells, and HEC-1A cells were selected for subsequent experiments. The result of CCK-8 showed that compared with the negative control group, the ability of HEC-1A cells in the miR-3653-3p group decreased on the 2nd, 3rd, 4th, and 5th days (all P < 0.05). The result of the Transwell chamber invasion test showed that the number of HEC-1A cell invasion after culturing for 26 h in the negative control group and the miR-3653-3p group was (80±11) and (21±4), respectively, and the difference was statistically significant ( t = 5.18, P < 0.01); compared with the negative control group, the number of cell invasion in the miR-3653-3p group decreased. The miRGator database was used to predict that the target gene of miR-3653-3p might be placenta-specific protein 8 (PLAC8). The relative expression levels of PLAC8 mRNA in HEC-1A cells in the negative control group and miR-3653-3p group were (6.26±0.83) and (0.97±0.31), respectively, and the difference was statistically significant ( t = 6.00, P < 0.01); the relative expression level of PLAC8 mRNA in the miR-3653-3p group was lower than that in the negative control group. Compared with the negative control group, the PLAC8 protein of HEC-1A cells decreased, and the expression of Wnt-β-catenin signaling pathway related proteins β-catenin, transforming growth factor β (TGF-β), GSK-3β, and Rac1 decreased in the miR-3653-3p group. Conclusions:miR-3653-3p may inhibit the proliferation and invasion of endometrial cancer cells by regulating the PLAC8-Wnt-β-catenin signaling pathway.
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Cancer immunotherapy has become a new generation of anti-tumor treatment, but its indications still focus on several types of tumors that are sensitive to the immune system. Therefore, effective strategies that can expand its indications and enhance its efficiency become the key element for the further development of cancer immunotherapy. Natural products are reported to have this effect on cancer immunotherapy, including cancer vaccines, immune-check points inhibitors, and adoptive immune-cells therapy. And the mechanism of that is mainly attributed to the remodeling of the tumor-immunosuppressive microenvironment, which is the key factor that assists tumor to avoid the recognition and attack from immune system and cancer immunotherapy. Therefore, this review summarizes and concludes the natural products that reportedly improve cancer immunotherapy and investigates the mechanism. And we found that saponins, polysaccharides, and flavonoids are mainly three categories of natural products, which reflected significant effects combined with cancer immunotherapy through reversing the tumor-immunosuppressive microenvironment. Besides, this review also collected the studies about nano-technology used to improve the disadvantages of natural products. All of these studies showed the great potential of natural products in cancer immunotherapy.
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Objective:To observe the protective effect of Angong Niuhuang pill on brain function of rats with sepsis, explore its protective mechanism, and provide the experimental basis for clinical application of Angong Niuhuang pill in the treatment of sepsis-associated encephalopathy (SAE).Methods:Thirty male Sprague-Dawley (SD) rats were divided into sham operation group, sepsis model group and Angong Niuhuang pill group according to random number table method, with 10 rats in each group. The sepsis model was established by cecal ligation and puncture (CLP); rats in sham operation group received open and closed abdomen. The rats in the Angong Niuhuang pill group were given Angong Niuhuang pill (0.3 g/kg) by gastric irrigation daily for 3 days before CLP, and the drugs were administrated 12 hours after modeling again. After 24 hours of CLP, the neuroreflex scores were evaluated, white blood cell count (WBC), the levels of serum neuron-specific enolase (NSE) and S100β were detected. Then the brain tissue was harvested. After hematoxylin-eosin (HE) staining, the pathological changes of brain tissue were observed under the light microscope. The mRNA expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in brain tissue were detected by polymerase chain reaction.Results:Compared with the sham operation group, the total score of neuroreflex scores in the sepsis model group and the Angong Niuhuang pill group were significantly reduced (4.43±1.40, 6.57±1.90 vs. 9.40±0.84, both P < 0.05), WBC, serum NSE, S100β were significantly increased [WBC (×10 9/L): 8.07±1.32, 5.84±0.94 vs. 3.60±0.32; NSE (μg/L): 1.04±0.14, 0.61±0.07 vs. 0.16±0.04; S100β (ng/L): 255.624±30.25, 97.72±15.41 vs. 46.88±12.03, all P < 0.05], and the mRNA expressions of IL-6 and TNF-α in brain tissue were significantly increased [IL-6 mRNA (2 -ΔΔCt): 5.668±2.195, 3.605±1.014 vs. 0.997±0.329; TNF-α mRNA (2 -ΔΔCt): 18.996±0.913, 1.746±0.710 vs. 0.674±0.132, all P < 0.05]. Compared with the sepsis model group, the total score of neuroreflex scores in the Angong Niuhuang pill group was significantly increased (6.57±1.90 vs. 4.43±1.40, P < 0.05), WBC, serum NSE, S100β concentration, and the mRNA expressions of IL-6 and TNF-α in the brain were significantly reduced [WBC (×10 9/L): 5.84±0.94 vs. 8.07±1.32, NSE (μg/L): 0.61±0.07 vs. 1.04±0.14, S100β (ng/L): 97.72±15.41 vs. 255.62±30.25, IL-6 mRNA (2 -ΔΔCt): 3.605±1.014 vs. 5.668±2.195, TNF-α mRNA (2 -ΔΔCt): 1.746±0.710 vs. 18.996±0.913, all P < 0.05]. Brain histopathological observation showed that the hippocampal neurons in the sepsis model group were disordered arrangement, a large number of neuronal nuclei were contracted, and the tissue was loose with obvious edema. Compared with the sepsis model group, the Angong Niuhuang pill group had less nuclear shrinkage and tissue edema. Conclusions:The pretreatment of the Angong Niuhuang pill can improve the brain dysfunction of septic rats and reduce the expression of pro-inflammatory cytokines in the brain. It is speculated that the Angong Niuhuang pill can protect the brain function in sepsis by inhibiting the inflammatory reaction in the brain.
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Growth arrest specific protein 6 (GAS6) plays an important role in the occurrence and development of tumors, and its signal transduction is involved in cell proliferation, adhesion and migration, but its related functions and molecular mechanisms in endometriosis (EMs) are still unclear. In this study, we searched and downloaded the transcriptome datasets of EMs from GEO database and performed GEO online analysis, and then screened out the differentially expressed genes and performed cluster analysis based on GO and KEGG pathway. The mRNA levels of the differentially expressed genes shared by more than three datasets were verified by qRT-PCR in the endometrium of ten women with no endometriosis and no clear disease and the ectopic endometrium of 11 patients with ovarian chocolate cysts. Immunohistochemistry and qRT-PCR were used to verify the expression of GAS6 and epithelial mesenchymal transition (EMT) marker genes, and immunofluorescence was used to co-label GAS6 and E-cadherin in endometriosis clinical samples. In this study, a total of 47 differentially expressed genes were screened out of the four transcriptome datasets, which were mainly enriched in processes such as cell migration and related signal pathways such as MAPK, PI3K-AKT, and tight junction. The mRNA levels of the nine differentially expressed genes shared by more than three datasets in endometriosis patients were consistent with the results of bioinformatics analysis. GAS6 expression levels in ectopic endometrium of EMs patients are higher than the control group (P < 0. 05), and EMs patients have the characteristics of EMT in the ectopic endometrial tissue, that is, the expression of E-cadherin is down-regulated (P < 0. 05) and the expression of vimentin is up-regulated (P < 0. 01). The expression of E-cadherin in the ectopic endometrial glandular epithelial cells of EMs patients is low while the expression of GAS6 is up-regulated, suggesting that GAS6 may mediate the EMT process in endometriosis. In conclusion, this study reveals that GAS6 is highly expressed in endometriosis patients and may mediate the EMT process to participate in the occurrence and development of endometriosis, providing a potential target for clinical treatment of endometriosis.
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OBJECTIVE@#To investigate the role of growth arrest-specific protein 6 (Gas6) in the process of the migration and osteogenic differentiation of human periodontal ligament cells (hPDLCs).@*METHODS@#After different concentrations of recombinant human Gas6 (rhGas6) were added to hPDLCs, cell prolife-ration experiment (CCK-8) was taken to observe the effect of rhGas6 on hPDLCs cell proliferation. Scratch test and cell migration test (Transwell) were taken to analyze the migratory ability of hPDLCs in different concentrations of rhGas6 groups. After osteogenic induction, real-time quantitative polymerase chain reaction (real-time PCR) was taken to detect the expression of the Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP). ALP staining was used to detect the amount of mineralized nodules.@*RESULTS@#After adding different concentrations of rhGas6, there were no statistically significant differences in hPDLCs cell proliferation among the experimental groups and the control group at 24, 48 and 72 hours (P>0.05). After 24 h of scratch, the healing area in the 800 μg/L of the rhGas6 group was greater than that in the control group, but without statistically significant difference (31.06%±13.70% vs. 21.79%±9.51%, P>0.05). In the migration test, after 24 h, the number of hPDLCs cells which penetrated through the membrane in the 800 μg/L rhGas6 group was significantly higher than that in the control group (P < 0.01). After rhGas6 was added and osteogenic induction, Runx2 and ALP gene expressions of hPDLCs in the 800 μg/L group were significantly higher than those in the control group (1.60±0.30 vs. 0.91±0.10, 2.81±0.61 vs. 0.86±0.12, P < 0.01). After Gas6 was knocked down, the ALP expression of hPDLCs was significantly lower than that of the control group (0.39±0.07 vs. 0.92±0.14, P < 0.01). There was no significant change in Runx2 expression (P>0.05). After 7 days of osteogenic induction, the mineralized nodules formed in the Gas6 knockdown group were significantly less than those in control group (0.25±0.04 vs. 1.00±0.11, P < 0.001). After 14 days of induction, the staining degree of the Gas6 knockdown group was lower than that of the control group, but there was no significant difference (0.86±0.04 vs. 1.00±0.16, P>0.05).@*CONCLUSION@#After downregulation of Gas6 gene, mineralized nodule formation was reduced and ALP gene expressions were decreased in the early stage of osteogenic induction (7 days). After addition of rhGas6, Runx2 and ALP gene expressions were increased and the number of cell migration was increased, suggesting that Gas6 might play a promoting role in the migration and osteogenic differentiation of human periodontal ligament cells.
Subject(s)
Humans , Alkaline Phosphatase , Cell Differentiation , Cell Proliferation , Cells, Cultured , Intercellular Signaling Peptides and Proteins , Osteogenesis , Periodontal LigamentABSTRACT
Objective • To evaluate the changes of urinary angiotensinogen (UAGT), fibroblast-specific protein-1 (FSP-1) and thrombin in the children with Henoch-Schonlein purpura nephritis (HSPN). Methods • Fourteen children with HSPN (HSPN group), 28 children with Henoch-Schonlein purpura (HSP) but without renal injury (HSP group) and 23 children with normal urinalysis (control group) were included in the study. Ten HSPN children before treatment (untreated group), 9 HSPN children after glucocorticoid (GC) pulse therapy (GC group) and 8 HSPN children after GC and cyclophosphamide (CTX) double pulse therapy (GC+CTX group) were also selected in the study. Clinical information and fresh morning urine samples were collected from all the children. UAGT, FSP-1 and thrombin in urine were measured by kits. Urine creatinine (Ucr) was also measured for correction. Results • UAGT/ Ucr and FSP-1 in HSPN group were significantly higher than those in HSP group and control group (P0.05), but thrombin levels in HSPN group and HSP group were both significantly higher than that in control group (P<0.05). UAGT/Ucr in HSPN untreated group had no significant difference, compared with GC group, while it was significantly higher than that in GC+CTX group (P=0.000). FSP-1 in untreated group was significantly higher than that in GC group, but had no significant difference, compared with GC+CTX group. There was no significant difference in thrombin among the 3 HSPN groups. Conclusion • UAGT/Ucr, FSP-1 and thrombin all increase in the urine of HSPN children, and UAGT/Ucr and FSP-1 may reflect the treatment effect to some extent. [Key words]
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INTRODUCTION: Cardiovascular disease is the main cause of mortality and morbidity in chronic renal failure. It's known that vascular calcification (VC) and carotid intima media thickness (CIMT) are strongly associated with cardiovascular diseases. Growth arrest specific protein 6 (Gas6) is a vitamin K-dependent protein and regulates various processes such as proliferation, cell survival, migration and inflammation. Gas6 is known to protect endothelial cells and vascular smooth muscle cells against apoptosis by inhibiting Bcl-2 induced Caspase 3 activation. The relationship between Gas6 and cardiovascular diseases has been demonstrated in many mouse models and cell cultures. However, there are conflicting reports whether Gas6 levels are increasing or decreasing in human studies of diabetic and/or chronic renal failure. In present study the aim was to examine plasma Gas6 levels and its relation with CIMT and coronary artery calcification score (CACS) in chronic kidney disease (CKD) patients. METHODS: Total of 137 patients of which 32 chronic hemodialysis and 105 predialysis patients as well as 73 healthy controls were enrolled in the study. Human Gas6 levels in serum samples were studied by ELISA method. CIMT was measured by ultrasonography. CACS was measured by multislice computed tomography. RESULTS: The mean age was 54.37±16.61 years in dialysis group, 55.20±14.80 years in predialysis group and 53.26±9.04 years in control group. Serum creatinine was 0.78±0.16 mg/dl in the control group and 1.96±1.64 mg/dl in the predialysis group and 5.94±1.55 mg/dl in the dialysis group. 24 hours urine protein levels were significally higher in the dialysis group than the predialysis and the control group. CIMT values were similar in predialysis and dialysis groups. These values were significantly higher than the control group. Although CACS was higher in dialysis group than predialysis and control group, the results were not statistically significant since the distribution range was very wide. Gas6 was 98.84±53.32 ng/mL in the control group and statistically higher than the dialysis (63.85±38.92 ng/mL) and the predialysis groups (54.96±38.49 ng/mL) (p=0.001). Gas6 levels were lower in diabetic patients than non-diabetics (53.69±35.26 ng/mL, 69.26±47.50 ng/mL, p=0.023, respectively). Negative correlation was detected between Gas6 and age, BMI, CACS, carotid IMT and proteinuria. In the logistic regression analysis, Gas6 remained significantly associated with BMI, CIMT and proteinuria. CONCLUSION: In our study, a negative correlation of Gas6 with BMI, CACS, CIMT and proteinuria and lower Gas6 levels in diabetic patients support that decreased Gas6 levels in chronic renal failure may have a role in vascular calcification through altered glucose tolerance, chronic inflammation, endothelial dysfunction and increased apoptosis. Our study has an importance because it is the first study showing a relation between Gas6 and proteinuria, CACS and carotid IMT in patients with chronic renal failure
INTRODUCCIÓN: La enfermedad cardiovascular es la principal causa de mortalidad y morbilidad en la insuficiencia renal crónica. Se sabe que la calcificación vascular (CV) y el grosor de la íntima-media de la carótida (CIMT, por sus siglas en inglés) están vinculados de forma muy estrecha con enfermedades cardiovasculares. La proteína específica del gen 6 de la detención de crecimiento (Gas6) es una proteína dependiente de la vitamina K y regula diversos procesos, como la proliferación, la supervivencia celular, la migración y la inflamación. La proteína Gas6 es conocida por proteger las células endoteliales y las células musculares lisas vasculares contra la apoptosis mediante la inhibición de la activación de la caspasa-3 inducida por la proteína Bcl-2. Se ha demostrado la relación entre la Gas6 y las enfermedades cardiovasculares en muchos modelos de ratones y cultivos celulares. Sin embargo, existen informes contradictorios acerca de si los niveles de Gas6 aumentan o disminuyen en estudios de humanos con insuficiencia renal crónica y/o diabética. En este estudio, el objetivo fue examinar los niveles plasmáticos de Gas6 y su relación con el CIMT y la puntuación de calcificación de las arterias coronarias (CACS, por sus siglas en inglés) en pacientes con enfermedad renal crónica (ERC). MATERIAL Y MÉTODOS: Un total de 137 pacientes fueron incluidos en el estudio, de los cuales 32 estaban en hemodiálisis crónica, 105 en prediálisis, y 73 pacientes representaban controles sanos. Se esudiaron los niveles de Gas6 en muestras de suero mediante el método ELISA. El CIMT se midió por medio de ecografía. La CACS se midió mediante tomografía computarizada multicorte. RESULTADOS: La edad media fue de 54,37 ± 16,61 años en el grupo de diálisis; 55,20 ± 14,80 años en el grupo de prediálisis, y 53,26 ± 9,04 años en el grupo de control. La creatinina sérica fue de 0,78 ± 0,16 mg/dl en el grupo de control; 1,96 ± 1,64 mg/dl en el de prediálisis, y 5,94 ± 1,55 mg/dl en el de diálisis. Las concentraciones de proteína en orina de 24 horas fueron significativamente más altas en el grupo de diálisis que en los de prediálisis y control. Los valores del CIMT fueron similares en los grupos de prediálisis y de diálisis. Estos valores fueron considerablemnete más altos que en el grupo de control. Aunque la CACS fue más alta en el grupo de diálisis que en los otros dos, los resultados no fueron estadísticamente significativos, ya que el rango de distribución fue muy amplio. La proteína Gas6 fue de 98,84 ± 53,32 ng/ml en el grupo de control y estadísticamente más alta que en los grupos de diálisis (63,85 ± 38,92 ng/ml) y de prediálisis (54,96 ± 38,49 ng/ml) (p = 0,001). Los niveles de Gas6 fueron más bajos en los pacientes diabéticos que en los no diabéticos (53,69 ± 35,26 ng/ml; 69,26 ± 47,50 ng/ml, [p = 0,023], respectivamente). Se detectó una correlación negativa entre la proteína Gas6 y la edad, el IMC, la CACS, el CIMT y la proteinuria. En el análisis de regresión logística, la Gas6 se mantuvo estrechamente relacionada con el IMC, el CIMT y la proteinuria. CONCLUSIÓN: En nuestro estudio, la correlación negativa de Gas6 con IMC, CACS, CIMT y proteinuria, y los niveles más bajos de Gas6 en pacientes diabéticos sustentan la idea de que la disminución de los niveles de Gas6 en la insuficiencia renal crónica puede jugar un papel en la calcificación vascular a través de la tolerancia alterada a la glucosa, la inflamación crónica, la disfunción endotelial y el aumento de la apoptosis. La importancia de nuestro estudio radica en que es el primero que muestra una relación entre la Gas6 y la proteinuria, la CACS y el CIMT en pacientes con insuficiencia renal crónica
Subject(s)
Humans , Vascular Diseases/complications , Calcinosis , Tunica Intima/abnormalities , Coronary Vessel Anomalies , Fibroblast Growth Factor 6/bloodABSTRACT
BACKGROUND: External quality assessment (EQA) is important for standardizing cancer biomarker assays, thereby, ensuring accurate and precise results. Although the human epididymis-specific protein 4 (HE4) assay has been increasingly used to detect and monitor ovarian malignancy in Korea, a nation-wide EQA program for HE4 has not been appropriately established. To conduct an EQA program, a large amount of quality control (QC) materials are required. This study aimed to produce HE4 QC materials for an EQA program and evaluate their homogeneity and stability. METHODS: QC materials for three different concentrations of HE4 were produced from the collected remnant sera of 275 patients for whom the HE4 assays were performed. These materials were evaluated for homogeneity between vials and stability during storage. The frozen QC materials were distributed to 13 representative organizations for a provisional EQA. RESULTS: The total coefficient of variation of the HE4 QC materials of three concentrations was 0.75%–1.24%, and no significant differences were noted between vials; therefore, the samples were considered to be homogenous. With respect to stability, the HE4 QC materials were found to be stable till 30 days when frozen and for 24 hours when refrigerated. The results of the provisional HE4 EQA were reviewed and the survey results were reported to each participant. CONCLUSIONS: The HE4 QC materials produced from remnant specimens were found to be homogenous between vials and stable in a frozen condition until 30 days. The findings of this study may be practically applied for establishing a future HE4 EQA program.
Subject(s)
Humans , Male , Epididymis , Korea , Ovarian Neoplasms , Quality ControlABSTRACT
OBJECTIVE@#To explore the role of transforming growth factor-β1/integrin-linked kinase/fibroblast-specific protein 1 (TGF- β1/ILK/FSP1) signaling pathway in cyclosporine A (CsA)-induced renal tubular epithelial cell transdifferentiation.@*METHODS@#Rat renal tubular epithelial NRK-52E cells were induced with 1 mg/L CsA, treated with TGF-β1 inhibitor (SB431542, 10 μmol/L), or transfected with the ILK-RNAi lentiviral expression vector (ILKshRNA) or a negative control vector before CsA induction. The expressions of TGF-β1, ILK and FSP-1 mRNAs and proteins in the cells were detected using real-time PCR and Western blotting. The positive cells for α-SMA expression were detected by immunohistochemistry.@*RESULTS@#Compared with the blank control cells, the cells treated with CsA showed significantly increased levels of TGF-β1, ILK and FSP-1 mRNAs and proteins ( < 0.05). The expressions of TGF-β1, ILK and FSP-1 were significantly lower in TGF-β1 inhibitor group than in CsA group ( < 0.05). The levels of ILK and FSP-1 were significantly decreased after shRNA-mediated ILK silencing ( < 0.05). The number of positive cells for -SMA was significantly lower in cells treated with SB431542 and in cells with ILK silencing than in the cells treated with CsA alone ( < 0.05).@*CONCLUSIONS@#The activation of TGF-β1/ILK/FSP-1 signaling pathway is an important mechanism for CsA-induced transdifferentiation in rat renal tubular epithelial cells. ILK participates in CsA-induced epithelialmesenchymal transition of renal tubular epithelial cells.
Subject(s)
Animals , Rats , Calcium-Binding Proteins , Cells, Cultured , Cyclosporine , Epithelial Cells , Epithelial-Mesenchymal Transition , Protein Serine-Threonine Kinases , Signal Transduction , Transforming Growth Factor beta1ABSTRACT
OBJECTIVE@#To observe the effect of electroacupuncture (EA) on the expressions of growth arrest-specific protein 7 (Gas7) and nerve growth factor (NGF) in arcuate nucleus (ARC) of rats with focal cerebral ischemia and explore the potential action mechanism of EA in treatment of focal cerebral ischemia.@*METHODS@#A total of 50 SD rats were randomized into 4 groups, named a normal group ( =12), a sham-operation group ( =12), a model group ( =14) and an EA group ( =12). In the model group and the EA group, the thread embolization method was adopted to duplicate the model of the right middle cerebral arterial embolism. In the sham-operation group, the skin of the neck was opened and sutured without any other intervention. In the EA group, EA was applied to "Baihui" (CV 20) and "Zusanli" (ST 36) on the left side, once a day, 30 min each time, consecutively for 21 days, while there was no any intervention in the normal group, the sham-operation group and the model group. Using the immunohistochemistry (IHC) method and Western blot method, the expressions of Gas7 and NFG of ARC on the ischemic side were determined. Using Nissle staining, the morphological changes in ARC neurons were observed.@*RESULTS@#The results of Nissle staining showed that there was no significant change in the morphology of ARC neurons in the normal group and the sham-operation group. In the model group, the volume of neuron cells was atrophied obviously and the cells were arranged irregularly. In the EA group, the morphology of ARC neuron was similar to the normal group. The results of IHC and Western blot indicated that the expressions of immunoreactive neurons and protein of Gas7 and NGF in ARC of the rats in the model group were increased obviously as compared with the normal group and the sham-operation group and the expressions in the EA group were further enhanced as compared with the model group (all <0.05).@*CONCLUSION@#Gas7 and NGF may be participated in the compensatory process of partial protection of the body in the patients with focal cerebral ischemia. EA up-regulates the expressions of Gas7 and NGF in ARC, which may be one of the neuroprotective mechanisms of EA in treatment of cerebral ischemia.
Subject(s)
Animals , Humans , Rats , Brain Ischemia , Metabolism , Therapeutics , Cerebral Infarction , Metabolism , Therapeutics , Electroacupuncture , Nerve Growth Factor , Metabolism , Nerve Tissue Proteins , Metabolism , Rats, Sprague-DawleyABSTRACT
@#Periodontal disease is one of the most common oral diseases and a prime reason for adult tooth loss. One of the goals of periodontal therapy is to regenerate and restore the periodontal tissues affected by periodontitis to their original architectural form and function. Cementum is an important component of periodontal tissue that plays a significant role in periodontal tissue regeneration. Explorations of the characteristics and components of cementum and the potential of growth factors related to cementum formation, especially the roles played by cementum-specific proteins and their potential in promoting periodontal regeneration, have become popular in current research. Here, we review the characteristics and components of cementum and the potential roles played by cementum-specific proteins during cementum formation and periodontal regeneration.
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Objective To evaluate the efficacy and safety of selective brain hypothermia (SBH) in the treatment of neonates with moderate or severe neonatal hypoxic-ischemic encephalopathy (HIE), and the effect of SBH treatment on serum levels of neuron-specific enolase (NSE) and central nervous specific protein S100. Methods A prospective randomized controlled trial was conducted. From January 2015 to June 2017, 42 children with moderate to severe HIE in the neonatal intensive care unit (NICU) of the First Affiliated Hospital of Bengbu Medical College were enrolled, and they were randomly divided into SBH treatment group and routine treatment group after obtaining the consent of the guardian of the children. The children in routine treatment group were given the traditional symptomatic supportive treatment, supplemented by drugs to promote nerve cell growth. On the basis of traditional treatment, the children in the SBH treatment group were given SBH treatment within 6 hours after birth. The nasopharyngeal temperature was maintained at 33.0-34.5 ℃ and the rectal temperature was maintained at 34.5-35.0 ℃. The general clinical data of the two groups including gender, gestational age, birth weight, age, 5-minute neonatal asphyxia score (Apgar score), score for neonatal acute physiology perinatal extension version Ⅱ (SNAPPEⅡ) were collected. The primary outcomes were hospitalized death, severe disability at 15 months of age, neonatal behavioral neurological assessment (NBNA) score at 28 days of age, and Bayley scales of infant development (BSID) score [including mental development index (MDI) score and psychomotor development index (PDI) score] at 15 months of age at follow-up. The secondary outcomes were serum levels of NSE and S100 protein. The occurrences of adverse events in the two groups were recorded. Results Among 42 HIE children, 1 child of severe congenital malformation and 1 child of platelet count (PLT)﹤50×109/L were excluded, and 40 children were enrolled in the study group. During the follow-up period, 2 children of SBH treatment group and 2 children of routine treatment group were lost or the outcome was unknown. Finally, 18 children of each group were enrolled in the analysis. There was no significant difference in the baseline data of gender, gestational age, birth weight, age, 5-minure Apgar score or SNAPPEⅡ score between the two groups, indicating that the baseline data of the two groups were balanced and comparable. The incidence of severe disability in the SBH treatment group was significantly lower than that in the routine treatment group [5.6% (1/18) vs. 44.4% (8/18), P﹤0.05]. There was 1 child death in the routine treatment group and no death in the SBH treatment group. Compared with the routine treatment group, the 28-day NBNA score of the SBH treatment group was increased by 2.9 [95% confidence interval (95%CI) = 1.0-4.8], BSID score at 15 months of age was improved significantly, MDI score was increased by 11.8 (95%CI = 4.3-19.3), and PDI score was increased by 12.4 (95%CI = 2.5-22.3), with significant differences between the two groups (all P﹤0.05). After 3 days of treatment, the serum NSE and S100 protein levels in both groups were significantly decreased as compared with those before treatment [NSE (μg/L): 30.15±15.18 vs. 31.32±14.75, S100 (ng/L): 387.5 (273.3, 573.0) vs. 890.0 (590.5, 1 162.5) in routine treatment group; NSE (μg/L): 29.09±16.22 vs. 32.25±15.43, S100 (ng/L): 402.5 (302.2, 580.5) vs. 842.0 (462.3, 1 200.5) in SBH treatment group, all P﹤0.05]. There was no significant difference in serum NSE or S100 protein level between the two groups (all P﹥0.05). There was no serious adverse event such as arrhythmia, large vein thrombosis or irreducible hypotension in both groups, and there was no significant difference in the incidence of general adverse events such as sinus bradycardia, scleredema, blood glucose disorder, or systemic infection between the two groups [16.7% (3/18) vs. 11.1% (2/18), 5.6% (1/18) vs. 5.6% (1/18), 22.2% (4/18) vs. 11.1% (2/18), 5.6% (1/18) vs. 5.6% (1/18), all P﹥0.05]. Conclusions SBH treatment could significantly increase the NBNA score at 28 days of birth and BSID score at 15 months of age, reduce the incidence of severe disability in moderate and severe HIE children, but it was not be proved that SBH could reduce the mortality. Compared with routine treatment, SBH treatment had no significant superiority on improving the levels of serum NSE and S100 protein, suggesting that SBH could not protect the brain by inhibiting the apoptosis of nerve cells and promoting the repair of nerve cells.
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Objective To investigate the diagnostic value of neuron-specific enolase(NSE),central nervous system specific protein (S100β),interleukin-6 (IL-6) in sepsis-associated encephalopathy (SAE).Methods Clinical data of patients admitted to ICU and diagnosed with sepsis were collected from January 2015 to June 2016 in Xiangya Hospital,Central South University.SAE was defined as cerebral dysfunction in the presence of sepsis that also fulfilled the exclusion criteria.The acute physiology and chronic health score (APACHE Ⅱ),sequential organ failure assessment (SOFA),NSE,S100β,IL-6,ICU stay time and 28-day mortality were compared between the two groups.NSE,S1003 and IL-6 were measured on the 1 st and 3rd day in ICU to determine the optimal cut-off value of SAE.Results Among 59 enrolled patients,36 were assigned to SAE group while 23 were non-SAE group.The SAE group had a significantly higher APACHE Ⅱ and SOFA scores,as well as the length of ICU stay (P ＜ 0.01).The levels of NSE,S1003 and IL-6 in the two groups both increased on the 1st day,and decreased on the 3rd day.The level of NSE on the 1st day [19.28 (13.00,30.52) μg/L vs 16.61 (7.58,22.01 μg/L)] and the 3rd day[16.03 (9.40,21.29) μg/L vs 11.39(8.49,15.00) μg/L,P=0.029],IL-6 on the 1st day[676.25(81.34,5 000.00) mg/L vs [209.10(42.27,648.20) mg/L,P =0.005] and the 3rd day [157.10 (72.85,687.63) mg/L vs 55.92 (31.62,177.00) mg/L,P =0.026] of SAE group was significantly higher than those of non-SAE group.However S100β between groups on the 1st day [0.33(0.15,0.54) μg/L vs 0.23(0.16,0.53) μg/L] and the 3rd day[0.19(0.10,0.29) μg/L vs 0.10(0.05,0.17) μg/L] was neither significant (P ＞0.05).The diagnostic values for SAE of NSE,S1003 and IL-6 were 14.36 μg/L,0.14 μg/L and 91.305 mg/L with sensitivity 61.1％,61.1％,72.2％ and specificity 73.9％,69.6％,69.6％,respectively.The diagnostic AUC of NSE and IL-6 combination was 0.774,95％ CI 0.651-0.896.Conclusion All sepsis patients have different degrees of brain injury.NSE combined with IL-6 on the 3rd day in ICU demonstrates the diagnostic significance of SAE.
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Objective To investigate the diagnostic value of neuron-specific enolase(NSE),central nervous system specific protein (S100β),interleukin-6 (IL-6) in sepsis-associated encephalopathy (SAE).Methods Clinical data of patients admitted to ICU and diagnosed with sepsis were collected from January 2015 to June 2016 in Xiangya Hospital,Central South University.SAE was defined as cerebral dysfunction in the presence of sepsis that also fulfilled the exclusion criteria.The acute physiology and chronic health score (APACHE Ⅱ),sequential organ failure assessment (SOFA),NSE,S100β,IL-6,ICU stay time and 28-day mortality were compared between the two groups.NSE,S1003 and IL-6 were measured on the 1 st and 3rd day in ICU to determine the optimal cut-off value of SAE.Results Among 59 enrolled patients,36 were assigned to SAE group while 23 were non-SAE group.The SAE group had a significantly higher APACHE Ⅱ and SOFA scores,as well as the length of ICU stay (P ＜ 0.01).The levels of NSE,S1003 and IL-6 in the two groups both increased on the 1st day,and decreased on the 3rd day.The level of NSE on the 1st day [19.28 (13.00,30.52) μg/L vs 16.61 (7.58,22.01 μg/L)] and the 3rd day[16.03 (9.40,21.29) μg/L vs 11.39(8.49,15.00) μg/L,P=0.029],IL-6 on the 1st day[676.25(81.34,5 000.00) mg/L vs [209.10(42.27,648.20) mg/L,P =0.005] and the 3rd day [157.10 (72.85,687.63) mg/L vs 55.92 (31.62,177.00) mg/L,P =0.026] of SAE group was significantly higher than those of non-SAE group.However S100β between groups on the 1st day [0.33(0.15,0.54) μg/L vs 0.23(0.16,0.53) μg/L] and the 3rd day[0.19(0.10,0.29) μg/L vs 0.10(0.05,0.17) μg/L] was neither significant (P ＞0.05).The diagnostic values for SAE of NSE,S1003 and IL-6 were 14.36 μg/L,0.14 μg/L and 91.305 mg/L with sensitivity 61.1％,61.1％,72.2％ and specificity 73.9％,69.6％,69.6％,respectively.The diagnostic AUC of NSE and IL-6 combination was 0.774,95％ CI 0.651-0.896.Conclusion All sepsis patients have different degrees of brain injury.NSE combined with IL-6 on the 3rd day in ICU demonstrates the diagnostic significance of SAE.
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No presente estudo, utilizou-se a melatonina e a proteína específica do oviduto (pOSP) nos meios de maturação in vitro. Foram avaliadas a expansão do complexo cumulus-ovócito (CCOs), as concentrações intracelulares de espécies reativas de oxigênio (ROS) e o desenvolvimento embrionário nos diferentes grupos (C = controle; T1 = somente com melatonina; T2 = com melatonina e pOSP e T3 somente com pOSP). No tocante à expansão do CCOs, houve diferença (P<0,05) dos valores obtidos no grupo C em relação aos valores médios dos grupos T1, T2 e T3, porém não houve diferença entre os valores obtidos nos tratamentos (P>0,05). Na dosagem de ROS, não houve diferença entre os valores médios obtidos no grupo C (26,4±10,9) e o valor verificado no grupo T1 (23,4±7,8), porém no grupo T2 (21,3±9,7) o valor médio mostrou-se satisfatório em relação ao valor do grupo C. No entanto, o valor médio do grupo T3 (16,6±10,5) foi o que demonstrou resultado mais satisfatório quando comparado aos demais grupos (P<0,05). A produção de embriões foi avaliada por meio da taxa de clivagem. Não houve diferença (P >0,05) entre os valores obtidos entre o grupo C (48,9 %) e os valores verificados nos grupos T1 (51,5 %), T2 (50 %), T3 (57,7 %), nem destes entre si. Este estudo permitiu concluir que a proteína específica do oviduto recombinante e a melatonina foram eficientes em melhorar a expansão dos CCOs. Além disso, as células tratadas com pOSP mostraram-se com menor quantidade de ROS, podendo a pOSP ser considerada um antioxidante proteico.(AU)
The present study used melatonin and recombinant oviduct specific protein (pOSP) in in vitro maturation medium (IVM). The expansion of the cumulus-oocyte complexes (COCs), the intracellular concentrations of reactive oxygen species (ROS) and embryo development of the different groups were evaluated (C = control; T1 = melatonin; T2 = melatonin and pOSP and T3 = pOSP). Regarding the COCs expansion, the groups T1, T2 and T3 showed satisfactory results compared with group C (P<0.05), but there was no difference between treatments (P>0.05). In the ROS dosage, there was no difference between the mean values obtained in group C (26.4 ± 10.9) and group 1 (23.4 ± 7.8). However, in group 2 (21.3 ± 9.7), the average value was found to be satisfactory in relation group C. Despite that, the average value of treatment 3 (16.6 ± 10.5) was the most satisfactory result found compared to the other groups (P<0.05). The production of embryos was evaluated by cleavage rate, there was no difference between the values obtained in group C and the values recorded in groups T1 (51.5 %), T2 (50 %), T3 (57.7 %), and among them. This study showed that the pOSP and the melatonin were effective in the improvement of the expansion of COCs cells. In addition, the cells that were treated with pOSP presented a lower amount of ROS, allowing the pOSP to be considered a proteic antioxidant.(AU)
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Animals , Female , Embryonic Development , Fallopian Tubes/chemistry , Melatonin/administration & dosage , Swine , Antioxidants , Cleavage Stage, Ovum , Embryo Culture Techniques/veterinary , In Vitro Techniques/veterinaryABSTRACT
Objective To evaluate the detection performance of the cobas8000 c702 fully automatic biochemical analyzer for de‐tecting the second generation Roche urine trace albumin (ALBU2) .Methods (1) The precise evaluation :with the allowable error stipulated by CLIA 88 as the basis ,the requirements were the repeat precision <1/4TEa ,and intermediate precision <1/3TEa;(2) the linear range and the evaluation of the reportable range :the EP6‐A scheme was adopted ,and extend to calculate the average re‐covery rate of dilution ,the clinical reportable range was evaluated by the average dilution recovery of 90% -110% ;(3) the carry o‐ver pollution assessment :the carry over pollution of serum albumin on urine trace albumin detection was evaluated by the judgment standard of carry over pollution rate of 0 .5% ;(4)the methodological comparative analysis :with SIEMENS BN Ⅱas the reference system ,the Roche Cobas 8000 C702 and the BN2 results were performed the correlation contrastive analysis .Results The repeat precision :low concentration CV=1 .98% .high concentration CV=1 .64% ;intermediate precision :low concentration CV=4 .35% , high concentration CV=1 .20% ;the linear range verification :the measurement range 5 .6-413 .55 mg/L ;clinical reportable range :in the maximum diluted multiples of 30 times ,the clinical reportable range was 5 .6-12 406 .5 mg/L ;the carry over pollution rate :serum albumin (42 .6 g/L) on urine trace albumin(6 .9 mg/L) ,the carry over pollution rate was 0 .28% ;the indoor comparison :in the concentration within 200 mg/L ,the regression line was Y=0 .896 X+5 .049 ,the correlation coefficient r2 =0 .994 4 ,the system shift was passed at the medical decision level .When the specimen concentration within 201-413 .55 mg/L ,the regression line was Y=0 .848X-10 .44 ,the correlation coefficient r2 =0 .917 ,the system shift was not passed at the medical decision level .Conclusion The detection of the Roche ALBU2 in the Cobas 8000 C702 platform can meet the clinical needs ,the comparison among different instruments has difference in different concentration ranges ,therefore the independent reference ranges should be established ac‐cording to the each instrument system .
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Epidemiological studies have shown a high prevalence of low serum testosterone levels in men with cardiovascular disease. Moreover, the tyrosine kinase receptor Axl, the ligand of which is growth arrest-specific protein 6 (GAS6), is expressed in the vasculature, and serum GAS6 levels are associated with endothelial dysfunction and cardiovascular events. Testosterone regulates GAS6 gene transcription directly, which inhibits calcification of vascular smooth muscle cells and provides a mechanistic insight into the cardioprotective action of androgens. This study was designed to determine the correlation between serum GAS6 and testosterone levels in male patients with coronary heart disease (CHD). We recruited 225 patients with CHD and 102 apparently healthy controls. Serum concentrations of GAS6 and soluble Axl were quantified by an enzyme-linked immunosorbent assay. Levels of high-sensitivity C-reactive protein, testosterone, estradiol, and other routine biochemical markers were also measured. Testosterone decreased from 432.69 ± 14.40 to 300.76 ± 6.23 ng dl-1 (P < 0.001) and GAS6 decreased from 16.20 ± 0.31 to 12.51 ± 0.19 ng ml-1 (P < 0.001) in patients with CHD, compared with control subjects. Multiple linear regression analysis showed that serum testosterone and GAS6 levels were positively associated in male patients with CHD. Alterations in GAS6 levels may influence the development of CHD. Downregulation of GAS6/Axl signaling in the presence of low sex hormone levels during disease progression is a potential mechanism by which GAS6 affects CHD. This study provides novel results regarding the influence of sex hormones on serum GAS6 levels in patients with CHD.
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Objective To study the effects of berberine (BBR) on the gene mRNA expression of fat-specific protein 27 (FSP27) and PR domain containing 16 (PRDM16) signal pathway in visceral white adipose tissues (VWAT) from type 2 dia? betic (T2DM) Chinese hamsters, and explore the related mechanisms. Methods The obese insulin-resistant (OIR) hamster model was induced by high-fat diet, and T2DM hamster model was created by OIR hamster model injected with low-dose streptozotocin. The control group was fed with standard laboratory chow. After the induction, the hamsters were randomly di?vided into control, OIR, obese T2DM and BBR-treated T2DM groups. After nine-week BBR treatment, real-time quantita?tive PCR was used to measure the gene mRNA expression changes of VWAT FSP27 and PRDM16 signal pathway and their target genes from different groups. Results Compared with control group, the gene mRNA expressions of PRDM16, CtBP-1, CtBP-2, C/EBPβ, PPARγ, PGC1α, PGC-1β and brown adipose tissue-specific genes such as UCP-1, Cidea, Elovl3, PPARα, and Acox, Cpt1 and Acadm were decreased and that of FSP27 and white adipose tissue-specific genes including Resistin, MEST and Serpina3k were increased in VWAT in OIR and obese T2DM groups. BBR treatment down-regulated FSP27 expression, enhanced PRDM16 signal pathway, and induced the gene mRNA expression of brown adipose tissue-spe?cific genes in VWAT of obese T2DM group to develop browning gene phenotype of VWAT, and then improved fat-induced insulin resistance. Conclusion The decreased FSP27 expression and increased PRDM16 expression are involved in the molecular mechanisms of browning of visceral white adipose tissues induced by BBR, and which contributes to improve ab?normal lipids metabolism and fat-induced insulin resistance in VWAT by enhancing consumption of energy as heat to re?store VWAT function.
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BACKGROUND: Dermatofibroma (DF) comprises a heterogeneous group of mesenchymal tumors, with fibroblastic and histiocytic elements present in varying proportions. The cell of origin of DF has been investigated, but remains unclear. OBJECTIVE: The present study attempted to investigate the expression of leukocyte-specific protein 1 (LSP1), a marker of fibrocytes, in DF. Additionally, we evaluated the effectiveness of LSP1 in the differential diagnosis of DF from dermatofibrosarcoma protuberans (DFSP). METHODS: Immunohistochemical staining was performed on 20 cases of DF using antibodies against LSP1, CD68, and factor XIIIa (FXIIIa). In addition, the expression of LSP1 and FXIIIa was evaluated in 20 cases of DFSP. RESULTS: Eighteen of 20 cases (90%) of DF stained positive for LSP1, with variation in the intensity of expression. CD68 was positive in 10 cases (50%), and FXIIIa was expressed in all cases of DF. There were differences between the regional expression patterns of the three markers in individual tumors. In contrast, only 2 of 20 cases of DFSP expressed LSP1, and none of DFSP cases stained positive for FXIIIa. CONCLUSION: The LSP1-positive cells in DF could potentially be fibrocyte-like cells. FXIIIa and CD68 expression suggests that dermal dendritic cells and histiocytes are constituent cells of DF. It is known that fibrocytes, dermal dendritic cells and histiocytes are all derived from CD14+ monocytes. Therefore, we suggest that DF may originate from CD14+ monocytes. Additionally, the LSP1 immunohistochemical stain could be useful in distinguishing between DF and DFSP.
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Antibodies , Dermatofibrosarcoma , Diagnosis , Diagnosis, Differential , Factor XIIIa , Fibroblasts , Histiocytes , Histiocytoma, Benign Fibrous , Langerhans Cells , MonocytesABSTRACT
Objective To explore the change of function and expression of hematopoietic lineage cell specific protein-1 (HS1) and phosphorylated HS1 (p-HIS) and factors devoting to HS1 phosphorylation in platelet with sepsis.Methods Plasma with rich platelet was collected from 150 sepsis patients and 50 healthy subjects, and comparison of platelets adhesion and aggregation were detected by micro-pore method and platelet aggregation instrument.Meanwhile the ATP concentrations of washed platelet of two groups were detected by the kit to compare release reaction.And then total HS1 (t-HIS) and p-HS1 of platelet from two groups were compared by using western blot.Afterwards the specific inhibitors of Src and Syk were used to verify the HS1 activation regulated by Src and Syk in LPS-induced cell model.Results The significant differences were present between healthy subjects and sepsis patients in platelet counts, platelet distribution width (PDW) and mean platelet volume (MPV) (P ＜ 0.01).The data showed the sepsis patients had greater ability than healthy subjects in adhesion, aggregation and release reaction.Meanwhile the platelets of sepsis patients had higher concentration of t-HS1 and p-HS1 than healthy subjects, and the specific inhibitors of Src and Syk , PP2 and piceatannol, inhibited the increase in p-HS1 in LPS-induced cell model.Conclusions Function of platelet is closely related to HS1 in sepsis and it will be a target for sepsis therapy.