Anti-inflammatory and DNA Repair Effects of Astragaloside IV on PC12 Cells Damaged by Lipopolysaccharide

Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses.Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50%cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100µmol/L astragaloside IV for 24 h.The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change>2,P<0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1µmol/L astragaloside IV for 24 h(P>0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P<0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P<0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1µmol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P<0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which are associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.

EXPRESSION AND SIGNIFICANCE OF TUMOR INFILTRATING DENDRITIC CELLS IN RENAL CELL CARCINOMA

Objective: To study the expression of dendritic cells in human renal cell carcinoma and explore the cause, so to reveal the mechanism of escaping immune surveillance in RCC. Methods: The expressions of CD83+DCS, CD1a+DCS,VEGF and TGF-β1 in tumoral, peritumoral and normal kidney tissues of RCC in 30 cases were detected by immunohistochemistry using streptavidin/peroxidese(SP) Results: CD83+DCS were mainly located in the peritumoral areas; whereas CD1a+DCS、were mainly retained within the cancer nests. The number of CD83+DCS was inversely correlated with the clinical stage(P<0.05); but there were no significant correlations between the number of CD1a+DCS、and the clinical stage(P>0.05). The expressions of CD83+DCS and CD1a+DCS have significant difference between the tumoral, peritumoral and normal kidney tissues(P<0.001). The expression of VEGF and TGF-β1 were significantly lower in samples with highly infiltrating CD83+DCS(P<0.05); Whereas CD1a+DCS were not (P>0.05). Conclusion: DC has the tendency to gathering in tumor, but because of the immunosuppressive cytokins, for example VEGF and TGF-β1, inhibits the maturation of DC, there are less mature TIDCS(CD83+TIDCS) in the tumoral tissues, they are mainly located in the peritumoral areas. This may contribute to the mechanism of escaping immune surveillance in RCC.

Treatment of pyogenic liver abscess by surgical incision and drainage combined with platelet-rich plasma:A case report

BACKGROUND Pyogenic liver abscesses are insidious in the early stage.Some cases progress rapidly,and the patient’s condition can worsen and even become life-threatening if timely treatment is not provided.Surgery and prolonged antibiotic treatment are often required if the abscess is large and liquefied and becomes separated within the lumen.CASE SUMMARY We report a case of bacterial liver abscess with a poor outcome following pharmacological treatment,review the literature related to the use of platelet-rich plasma(PRP)in the treatment of hepatic impairment and partial hepatectomy in animals,and discuss the prognostic features of surgical incision and drainage combined with PRP in the treatment of bacterial liver abscesses.This is the first case describing the use of PRP in the treatment of a bacterial liver abscess in humans,providing new ideas for the treatment of this condition.CONCLUSION This case highlights the importance of surgical treatment for bacterial liver abscesses that are well liquefied and poorly managed medically.PRP may produce antimicrobial effects and promote the regeneration and repair of liver tissue.

The Clinical Evaluation of Minimally Invasive Treatment for Axillary Osmidrosis

Background: Traditional treatment of axillary odour is the surgical removal of skin and subcutaneous tissue along the edge of the axillary hair in a fusiform shape, which requires the restriction of upper limb activity, has large scars and a high recurrence rate. Minimally invasive methods have developed in recent years. The subcutaneous trimming method with small incision reduces the incision compared to traditional incision, but still requires skin incision, which does not achieve true, minimally invasive treatment. Simple negative pressure suction is difficult to destroy the sweat glands and sebaceous glands in the dermis layer. Thus, we hypothesized that combination of three minimally invasive methods might be more effective. Objective: To explore the clinical effect of combined treatment of axillary osmidrosis by three minimally invasive methods of microwave, scraping & suction and scratching. Methods: From October 2015 to October 2020, 80 patients with underarm odor were enrolled and lacal swelling anesthetic was applied. Microwave was used to treat sweat glands and sebaceous glands in the dermis following the pores. The curettage-aspiration was used to scrape and suck the apocrine sweat glands in the subcutaneous fat layer and the junction between the subcutaneous fat and dermis. To achieve the purpose of minimally invasive and thorough treatment of underarm odor, the spoon scraped the remaining sweat gland tissue. Results: Among 80 patients on 160 sides, all cases (100.00%) were cured on 160 sides at 1 month post-operative follow-up, 75 cases (93.75%) were cured on 150 sides, 5 cases (6.25%) on 5 sides were ineffective at 6-month post-operative follow-up, 74 cases (92.50%) were cured on 148 sides, 6 cases (7.50%) on 5 sides were ineffective at 1-year post-operative follow-up. Complications occurred in 5 cases (6.25%) after operation: 2 cases (2.50%) with skin necrosis on 3 sides, 1 case (1.25%) with uneven skin on 2 sides, 1 case (1.25%) with axillary abscess on 2 sides, and 1 case (1.25%) with keloids on 2 sides. Conclusion: The combination of three minimally invasive methods of microwave, scraping & suction and scratching to treat underarm odor is simple operation, has high cure rate, quick postoperative recovery, and few complications. It is an effective method for minimally invasive and thorough treatment of underarm odor in clinic.

Identification of microRNAs as novel biomarkers for esophageal squamous cell carcinoma: a study based on The Cancer Genome Atlas (TCGA) and bioinformatics

Background:MicroRNAs (miRNAs) have played important roles in the regulation of gene expression in many cancers,but their roles in esophageal squamous cell carcinoma (ESCC) are still unclear.The aim of this study was to determine the potential ESCC-specific key miRNAs from a large sample dataset in The Cancer Genome Atlas (TCGA).Methods:Integrative bioinformatics analysis was used to identify key ESCC-specific miRNAs related to the ESCC patients' tumor histological grade and lymphatic metastasis from TCGA.Next,these key miRNA potential gene regulatory functions and relationships with ESCC patients' clinical characteristics and overall survival were analyzed.Finally,three key miRNAs were selected randomly and quantificational real-time polymerase chain reaction (qRT-PCR) was used to validate in 51 newly diagnosed ESCC patients' tissues samples (collected from Nov.2017 to Feb.2019,in Wuwei,China) whether the bioinformatics analyses results were reliable and valid.Two-tailed Student's t test,Pearson Chi-squared test and Kaplan-Meier survival analysis were used in this study.Results:Thirty-five ESCC-specific miRNAs from TCGA database were investigated (fold-change > 2.0,P < 0.05),and 28 participated in the miRNAs-mRNAs co-expression network construction,while 17 were related with ESCC patients' tumor histological grade,TNM stage,and lymphatic metastasis (P < 0.05).Meanwhile,six miRNAs (including miR-200b-3p,miR-31-5p,miR-15b-5p,miR-141-3p,miR-135b-5p,and miR-195-5p) were correlated with overall survival of ESCC patients (log-rank,P < 0.05).MiR-135b-5p,miR-15b-5p,and miR-195-5p were selected for verification of the expression levels in 51 ESCC patients' tissue samples by using qRT-PCR.We found that the fold-changes between qRT-PCR and TCGA were completely consistent.The results also suggested that miR-135b-5p,miR-15b-5p,and miR-195-5p were significantly correlated with tumor differentiation degrees (P < 0.05),miR-195-5p was significantly correlated with tumor TNM stage (P < 0.05),and miR-135b-5p was significantly correlated with lymph-node metastasis (P < 0.05).MiR-135b-5p,miR-15b-5p,and miR-195-5p expression levels,ESCC patient clinical features association analysis results and the aforementioned TCGA bioinformatics analyses were similar.Conclusion:This study identified key ESCC-related miRNAs.The key miRNAs are worthy of further investigation as potential novel biomarkers for diagnosis,classification,and prognosis of ESCC.

Extractum trametes robiniophila murr protects murine podocyte via actin cytoskeleton stabilization

The Chinese medicine Extractum trametes robiniophila murr(ETRM)is the extract of a type of fungus.Recent studies have suggested that ETRM efficiently improves the effacement of podocyte foot processes in adriamycin(ADR)-induced nephrotic rats.In the present study,we aimed to assess whether ETRM modulated the actin rearrangements of podocytes and involved signaling molecules,includingα-actinin-4 and nephrin.Podocytes were treated with ADR(0.5μmol/L),ADR(0.5μmol/L)+dexamethasone(Dex)(1μmol/L),ADR(0.5μmol/L)+ETRM(10 mg/mL).The F-actin in the podocytes was stained by fluorescent phallotoxins and observed by confocal microscopy.The expression levels ofα-actinin-4 and nephrin were tested by real-time PCR and Western blotting analysis.The administration of ETRM could significantly prevent ADR-treated podocytes from actin rearrangement.Both ETRM and Dex could stabilize podocyte actin cytoskeletons.Moreover,α-actinin-4 might act as a potential target for ETRM functionality in podocyte actin rearrangements.However,pretreatment with ETRM could not inhibit the up-regulation of nephrin as a result of ADR treatment.ETRM could improve the cytoskeleton stability in ADR-induced actin rearrangement of podocytes via regulatingα-actinin-4 expression.

Mechanistic study of ventricular hook anchor for heart valve replacement or repair

Objective:The objective of this study was to investigate the mechanics of ventricular anchor for heart valve repair or replacement.Methods:Thirteen anchors were designed based on six geometric parameters of the anchor teeth:width,thickness,root length,radius of curvature,tip angle,and tip length.Finite element method was applied to simulate the process of the anchor compressing into a sheath.The Von-Mises strain,peak pulling force,and bite depth were evaluated.An experiment was performed to validate the simulation.Results:The maximum Von-Mises strain was at the contact region of the anchor in a sheath where the teeth were compressed against one another and were distorted.The maximum strain increased with an increase in tooth width,thickness,radius of curvature and tip angle.The peak pulling force increased as tooth thickness and width increased,and radius of curvature decreased.Both the radial and axial bite depths increased with an increase in the tip length at the tip length
3 mm.The radial bite depth increased with an increase in the radius of curvature.Conclusion:1)the maximum strain depends primarily on the tooth width,thickness,radius of curvature and tip angle;2)the peak pulling force depends primarily on the tooth width,thickness,radius of curvature;3)the axial bite depth depends primarily on the tip length at the tip length
3 mm.The radial bite depth depends on the radius of curvature and the tip length at the tip length>3 mm.The study provides guidance for ventricular anchor design.