Experience of implanting fiducial markers for 504 cases with body malignant solid tumors

Objective:The purpose of the study is to investigate the technical points,effects and complications of fiducial marker implantation within target areas before the CyberKnife treatment on body malignant solid tumors.Methods:Five hundred and four cases of patients with body malignant solid tumors accepted fiducial implantation within target areas under CT guidance before the treatment of CyberKnife.Observe the complications and effect.Results:Among the 504 cases,500 cases successfully accepted the implantation(a success rate of 99.2%).158 patients felt pain at the punctured sites and 3 patients had tachycardia.33 patients had abdominal pain after the surgery due to a small amount of bleeding in the needle passage during liver puncturing process.Among the 19 lung cancer patients who accepted lung paracentesis,1 case had light pneumothorax and 1 case got light haemothorax.Among the 453 patients who accepted liver paracentesis,6 had fiducial migration.Conclusion:The method of fiducial implantation within target areas before treating body malignant solid tumor with CyberKnife is minimally invasive and comparatively secure.

Protective effects of a composition of Chinese herbs- Gurigumu-13 on retinal ganglion cell apoptosis in DBA/2J glaucoma mouse model

AIM： To explore the concrete mechanism of a Mongolian compound medicine-Gurigumu-13 （GRGM） for glaucoma treatment. METHODS： DBA/2J mice, as glaucoma models, were intragastric administrated with GRGM to study the effect of GRGM on retinal ganglion cells （RGCs）. The loss of RGCs was evaluated with the number of RGCs and axons. The expression of the target protein of RGCs or mouse retinas was determined by Western blot. The relative content of malondialdehyde （MDA） was examined by ELISA assay. RESULTS： GRGM distinctly improved retina damage via increasing the number of neurons, RGCs and axons in a concentration dependent manner. Meanwhile, GRGM obviously decreased the high level of MDA and the expression of oxidative stress-related proteins in retinas of DBAJ2J mice, but promoted the expression of antioxidant proteins. Additionally, GRGM also significantly inhibited the protein expression of Bip and Chop, which were markers of endoplasmic reticulum stress-induced apoptosis. CONCLUSION： GRGM have obvious protective effects on RGCs in DBA/2J mice, and increase the number of RGCs and axons via inhibiting oxidative stress and endoplasmic reticulum stress.

EZH2 Contributes to Anoikis Resistance and Promotes Epithelial Ovarian Cancer Peritoneal Metastasis by Regulating m6A

Objective:Histone modification has a significant effect on gene expression.Enhancer of zeste homolog 2(EZH2)contributes to the epigenetic silencing of target chromatin through its roles as a histone-lysine N-methyltransferase enzyme.The development of anoikis resistance in tumor cells is considered to be a critical step in the metastatic process of primary malignant tumors.The purpose of this study was to investigate the effect and mechanism of anoikis resistance in ovarian adenocarcinoma peritoneal metastasis.Methods:In addition to examining EZH2 protein expression in ovarian cancer omental metastatic tissues,we established a model of ovarian cancer cell anoikis and a xenograft tumor model in nude mice.Anoikis resistance and ovarian cancer progression were tested after EZH2 and N6-methyladenosine(m6A)levels were modified.Results:EZH2 expression was significantly higher in ovarian cancer omental metastatic tissues than in normal ovarian tissues.Reducing the level of EZH2 decreased the level of m6A and ovarian cancer cell anoikis resistance in vitro and inhibited ovarian cancer progression in vivo.M6a regulation altered the effect of EZH2 on anoikis resistance.Conclusion:Our results indicate that EZH2 contributes to anoikis resistance and promotes ovarian adenocarcinoma abdominal metastasis by m6A modification.Our findings imply the potential of the clinical application of m6A and EZH2 for patients with ovarian cancer.

Differentially expressed genes analysis and target genes prediction of miR-22 in breast cancer

Objective miR-22 is highly active in breast cancer, especially in the luminal B and HER2 subtypes.However, the detailed potential of the use of target genes for miR-22 in breast cancer are still unclear. Inthis study, we aimed to discover potential genes and the miRNA-DEGs network of miR-22 in breast cancerusing bioinformatics approaches.Methods Analysis of microarray data GSE17508 (including 3 miR-22 knockout samples and 3 controls)obtained from the Gene Expression Omnibus (GEO) database was performed. Differentially expressedgenes (DEGs) between the miR-22 knockout samples and the three control samples were detected usingGEO2R. The gene ontology (GO) functional enrichment analysis and protein-protein interaction (PPI)network of DEGs were performed using the online tool Metascape and STRING database, separately. ThemiR-22 and DEG networks were obtained from the miRNet database. Cytoscape software was used toconstruct and analyze a merged miRNA-DEG network. The online tools database, mirDIP 4.1, was used topredict miR-22 target genes.Results Certain DEGs and miRNAs may be potential targets for predicting and treating miR-22 expressedbreast cancer.Conclusion We constructed a prognostic model of rectal adenocarcinomas based on four immunerelatedlncRNAs by analyzing the data based on TCGA database, with high prediction accuracy. We alsoidentified two biomarkers with poor prognosis (PXN-AS1 and AL158152.2) and one biomarker with goodprognosis (LINC01871).

Metabolomics of Fuzi-Gancao in CCl4 induced acute liver injury and its regulatory effect on bile acid profile in rats

BACKGROUND Fuzi(Radix aconiti lateralis)-Gancao(Radix glycyrrhizae)is one of the most classical drug pairs of traditional Chinese medicine.In clinical practice,decoctions containing Fuzi-Gancao(F-G)are often used in the treatment of liver diseases such as hepatitis and liver failure.AIM To investigate the metabolomics of F-G in CCl4 induced acute liver injury in rats and its regulatory effect on the bile acid profile.METHODS The pharmacodynamic effect of F-G on CCl4 induced acute liver injury in rats was evaluated,and an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the simultaneous determination of 92 metabolites from multiple pathways was established to explore the protective metabolic mechanism of F-G in serum on the liver.RESULTS Twenty-four differential metabolites were identified in serum samples.The primary bile acid biosynthetic metabolic pathway was the major common pathway in the model group and F-G group.Subsequently,a UPLC-MS/MS method for simultaneous determination of 11 bile acids,including cholic acid,ursodeoxycholic acid,glycochenodeoxycholic acid,glycochenodeoxycholic acid,taurocholic acid,glycocholic acid,chenodeoxycholic acid,deoxycholic acid,taurochenodeoxycholic acid,taurocholic acid,and glycinic acid,was established to analyze the regulatory mechanism of F-G in serum.F-G decreased the contents of these 11 bile acids in serum in a dose-dependent manner compared with those in the model control group.CONCLUSION F-G could protect hepatocytes by promoting the binding of free bile acids to glycine and taurine,and reducing the accumulation of free bile acids in the liver.F-G could also regulate the compensatory degree of taurine,decreasing the content of taurine-conjugated bile acids to protect hepatocytes.

Relationship between miR-7-5p expression and ^(125)I seed implantation efficacy in pancreatic cancer and functional analysis of target genes

Objective The aim of this study was to investigate the relationship between miR-7-5p expression and intertissue-^(125)I irradiation sensitivity in pancreatic cancer tissues and to analyze the function of target genes.Methods Thirty-seven patients with unresectable pancreatic ductal adenocarcinoma(PDAC)treated with radioactive ^(125)I seed implantation were enrolled.RT-PCR was used to detect the expression level of miR-7-5p in cancer tissues and analyze the relationship between miR-7-5p expression and ^(125)I radiation sensitivity.Bioinformatic software and online tools were used to predict the miR-7-5p target genes and analyze their functional annotation and pathway enrichment.Results Radioactive ^(125)I seed implantation was followed up for 2 months.The objective response rate of the miR-7-5p high expression group was 65.0%(13/20),whereas the objective response rate of the miR-7-5p low expression group was 5.88%(1/17),and the difference between the two groups was statistically significant(χ^(2)=13.654,P<0.001).A total of 187 target genes were predicted using three databases.GO functional annotation showed that target genes were mainly involved in cellular response to insulin stimulus,regulation of gene expression by genetic imprinting,cytosol,peptidyl-serine phosphorylation,bHLH transcription factor binding,cargo loading into vesicles,cellular response to epinephrine stimulus,and nucleoplasm.KEGG pathway enrichment analysis showed that target genes were mainly involved in the ErbB signaling pathway,HIF-1 signaling pathway,axon guidance,longevity regulatory pathway,endocrine resistance,glioma,choline metabolism in cancer,and EGFR tyrosine kinase inhibitor drug resistance.Molecular complex detection analysis by Cytoscape revealed that PIGH,RAF1,EGFR,NXT2,PIK3CD,PIK3R3,ERBB4,TRMT13,and C5orf22 were the key modules of miR-7-5p target gene clustering.Conclusion The expression of miR-7-5p in pancreatic cancer tissues positively correlated with the radiosensitivity of ^(125)I seeds.Via targeted gene regulation,miR-7-5p acts on the network of multiple signaling pathways in PDAC and participates in its occurrence and development.Thus,miR-7-5p may become a predictive index of ^(125)I seed implantation therapy sensitivity in PDAC patients.

Apoptosis of Glioblastoma U251 Cells Induced by Carmustine Combined All-trans Retinoic Acid via Regulating Cyclin E and p27kip 1

The effect and mechanism of carmustine（BCNU） combined with all-trans retinoic acid（ATRA） on the apoptosis of human glioblastoma U251 cells were investigated by means of 3-（4,5-dimethylthiazol-2-yl）-2,5-diphe- nyltetrazolium bromide（MTT） assay, flow cytometry, reverse transcription-polymerase chain reaction（RT-PCR） and Western blot analysis. The results show that BCNU or ATRA shows time- and dose-dependent inhibition effects on human glioblastoma U251 cells and the combination of BCNU with ATRA shows an synergistic inhibition effect on human glioblastoma U251 cells, and the combined BCNU and ATRA can significantly inhibit the proliferation of human glioblastoma U251 cells, and induce the apoptosis of them, making the cells arrest in the stage of G1 phase, the stage of S and G2 phases decline, the rate of the apoptosis of human glioblastoma U251 cells increase, the corresponding mRNA expression of cyclin E and cyclin-dependent kinase 2（CDK2） downregulated and the correspon- ding mRNA expression of p27kip 1 unregulated. In addition, the combined BCNU and ATRA reduced the protein expression of nuclear factor kappa B（NF-κB）. Taken together, these results suggest that the treatment of human glioblastoma U251 cells with a combination application of ATRA and BCNU can exert synergistic effect, the course of this kind of combination chemotherapy may likely be associated with multiple molecular mechanisms for apoptosis, furthermore, the cyclin E and p27kip 1 should be considered as novel targets for controlling the growth of glioblastoma cells.

The Present Scenario, Challenges, and Future Anticipation of Traditional Mongolian Medicine in China

The objective was to study the present scenario,challenges,and future anticipation of traditional Mongolian medicine(TMM)in China.The literature was retrieved using various sources of information such as meetings with TMM experts,research and review articles,national health databases,and government websites and authoritative papers.Health care using TMM,TMM education and research,and the pharmaceutical industry of TMM has been improved by implementing TMM growth policies.TMM has been extensively used for health care in Mongolia and in various regions of China including Beijing.The national education system of China contains a stream of TMM education.There is a yearly increase in research productivity in the field of TMM due to extensive research funding by the government.The rapidly growing pharmaceutical industry of TMM has provided more availability of TMM to more patients.The literature search revealed good preservation and development of TMM in China,urging the need to take steps for the promotion of its spread and development in China and worldwide.

Chinese Tuina Downregulates the Elevated Levels of Tissue Plasminogen Activator in Sciatic Nerve Injured Sprague-Dawley Rats

Objective：To elucidate the mechanism of Chinese tuina in treating sciatic nerve crush injury,and to detect the levels of tissue plasminogen activator（t PA）and plasminogen activator inhibitor-1（PAI-1）,which is thought to play an important role in nerve regeneration.Methods：Thirty-two adult male Sprague-Dawley rats were subjected to sciatic nerve crush injury and 16 rats（sham-operated group）went through a sham operation.Control group was given no treatment while tuina group received tuina therapy since day 7 post-surgery.Tuina treatment was performed once a day and lasted for 20 days.The sciatic functional index was examined every 5days during the treatment session.The rats＇gastrocnemius muscles were evaluated for changes in mass and immunohistochemistry techniques were performed to detect the levels of tP A and PAI-1.Results：Tuina therapy improved the motor function of sciatic nerve injured rats（P〈0.05）,however,it did not increase muscle volume（P〈0.05）.Tuina downregulated the levels of t PA and PAI-1（P〈0.05）.Conclusions：The present study implies that tuina treatment could accelerate rehabilitation of peripheral nerve injury.

Differences in Pathological Characteristics and Laboratory Indicators in Adult and Pediatric Patients with Henoch-Sch？nlein Purpura Nephritis

We aimed to investigate the differences in renal histopathological changes and laboratory parameters between adult and pediatric patients with Henoch-Sch？nlein purpura nephritis（HSPN）, and to analyze the correlation between laboratory parameters and renal histopathological grading. A total of 139 patients diagnosed with HSPN between September 2010 and December 2014 at the First Hospital of Jilin University, China, were retrospectively reviewed. The clinical and pathological characteristics were examined and compared between the adult and the pediatric patients. A majority of adult（75.0%） and pediatric（66.2%） patients were categorized as pathological grade Ⅲ HSPN. Adults having crescent lesions, interstitial fibrosis and renal artery involvement significantly outnumbered child counterparts（all P〈0.05）. Pathological grading showed a positive correlation with 24-h urine protein（r=0.307, P=0.009）, microalbuminuria（r=0.266, P=0.000） and serum globulin（r=0.307, P=0.014）, and a negative correlation with serum albumin（r=0.249, P=0.037） in pediatric patients with HSPN. Among adult patients with HSPN, histopathological grading showed a positive correlation with 24-h urine protein（r=0.294, P=0.015）, microalbuminuria（r=0.352, P=0.006）, α1-microglobulin（r=0.311, P=0.019） and immunoglobulin G（r=0.301, P=0.023） in urine, and serum creatinine（r=0.292, P=0.018）. Further, a negative correlation between serum albumin and pathological grading was also observed（r=0.291, P=0.018）. In conclusion, the severity of renal pathological lesions in HSPN patients is well reflected by the levels of proteinuria. Adult patients have more severe renal histopathological changes than pediatric patients.

Differential gene screening and functional analysis in docetaxel-resistant prostate cancer cell lines

Objective Docetaxel-based combination chemotherapy has traditionally been the standard treatment for metastatic castration-resistant prostate cancer(PCa).However,most patients eventually develop resistance to this treatment,which further reduces their survival.This study aimed to determine key molecular genes in docetaxel-resistant PCa cell lines using bioinformatic approaches.Methods The analysis of microarray data GSE33455(including DU-145/DU-145R and PC-3/PC-3R cell lines)obtained from the Gene Expression Omnibus(GEO)database was performed using GEO2R.Differentially expressed genes(DEGs)of DU-145/DU-145R and PC-3/PC-3R cell lines were selected,and the intersection of DEGs between the two groups was obtained.DEGs were annotated with the Gene Ontology(GO)function and enriched with the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway using an online platform(https://cloud.oebiotech.cn/task/detail/array_enrichment/).The online tool Search Tool for the Retrieval of Interacting Genes(https://string-db.org/)was used to obtain the DEG network graph and matrix list,which was imported into Cytoscape 3.6.1 and analyzed using the Molecular Complex Detection plug-in to detect potential functional modules in the network.Results A total of 131 intersection DEGs were identified between non-treated and docetaxel-resistant PCa cell lines.GO functional annotation showed that the main genes involved were present in the plasma membrane and were involved in positive regulation of ubiquitin-protein transferase activity,positive regulation of pseudopodium assembly,centriolar subdistal appendage,and heterophilic cell-cell adhesion via plasma membrane cell adhesion molecules.KEGG pathway enrichment analysis revealed that DEGs were mainly involved in IL-17 signaling pathway,cytokine-cytokine receptor interaction,rheumatoid arthritis,legionellosis,and folate biosynthesis.We identified two distinct hubs of DEGs:(1)CD274,C-X-C motif chemokine ligand(CXCL)1,DExD/H-box helicase 58,CXCL2,CXCL8,colony-stimulating factor 2,C-X-C motif chemokine receptor 4(CXCR4),CXCL5,and CXCL6 and(2)argininosuccinate lyase,argininosuccinate synthase 1,and asparagine synthetase.Except for the CXCR4 gene that was downregulated,the other 11 genes showed upregulated expression.Conclusion Certain differential genes may be potential targets for predicting and treating metastatic docetaxel-resistant PCa.

Th17 cells are involved in mouse chronic obstructive pulmonary disease complicated with invasive pulmonary aspergillosis

Background:The incidence of chronic obstructive pulmonary disease(COPD)complicated with invasive pulmonary aspergillosis(IPA)has increased in the last two decades.The mechanism underpinning susceptibility to and high mortality of COPD complicated with IPA is unclear,and the role of T helper cells 17(Th17 cells)in the compound disease remains unknown.Therefore,this study aimed to assess the function of Th17 cells in COPD combined with IPA.Methods:COPD,IPA,and COPD+IPA mouse models were established in male wild type C57/BL6 mice.The amounts of Th17 cells and retinoic acid-related orphan receptorsγt(RORyt)were tested by flow cytometry.Then,serum interleukin(IL)-17 and IL-23.levels were detected by enzyme-linked immunosorbent assay(ELISA)in the control,COPD,IPA and COPD+IPA groups.In addition,COPD+IPA was induced in IL-17 knockout(KO)mice,for determining the role of Th17 cells in COPD+IPA.Results:Compared with the COPD group,the COPD+IPA group showed higher amounts of blood RORyt([35.09±16.12]%vs.[17.92±4.91]%,P=0.02)and serum IL-17(17.96±9.59 pg/mL vs.8.05±4.44 pg/mL,P=0.02),but blood([5.18±1.09]%vs.[4.15±0.87]%,P=0.28)and lung levels of Th17 cells(1.98±0.83]%vs.[2.03±0.98]%,P=0.91),lung levels of RORyt([9.58±6.93]%vs.[9.63±5.98]%,P=0.49)and serum IL-23(51.55±27.82 pg/mL us.68.70±15.20 pg/mL,P=0.15)showed no significant differences.Compared with the IPA group,the COPD+IPA group displayed lower amounts of blood([5.18±1.09]%vs.[9.21±3.56]%,P=0.01)and lung Th17 cells([1.98±0.83]%vs.[6.29±1.11]%,P=0.01)and serum IL-23(51.55±27.82 pg/mL vs.154.90±64.60 pg/mL,P=0.01)and IL-17(17.96±9.59 pg/mL uUs.39.81±2.37 pg/mL,P=0.02),while comparable blood([35.09±16.12]%Vs.[29.86±15.42]%,P=0.25)and lung levels of RORγt(9.58±6.93]%VUS,[15.10±2.95]%,P=0.18)were found in these two groups.Finally,Aspergillus load in IL-17 KO COPD+IPA mice was almost 2 times that of COPD+IPA mice(1,851,687.69±944,480.43"vs.892,958.10±686,808.80,t=2.32,P=0.02).Conclusion:These findings indicate that Th17 cells might be involved in the pathogenesis of COPD combined with IPA,with L-17 likely playing an antifungal role.