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Objective:To explore the prognostic biomarkers of glioblastoma (GBM) in the tumor microenvironment (TME) and its function.Methods:A total of 169 GBM samples of 161 GBM patients were collected from the Cancer Genome Atlas (TCGA) database. ESTIMATE algorithm in R4.1.0 software was used to calculate the proportion of immune components and stromal components in TME, which were expressed as immune score and stromal score, respectively. According to the median value of the two scores, 169 GBM samples were divided into the high score group and the low score group, respectively, 84 each in each group (those whose scores were equal to the median were not involved in the grouping). The differentially expressed genes (DEG) [false discovery rate (FDR) < 0.05] between the high score group and the low score group of the two scores were obtained by using limma package, and the co-up-regulated and co-down-regulated DEG of the two scores were obtained by using Venn program. Based on the STRING database, the protein interaction (PPI) network of co-up-regulated and down-regulated DEG of immune score and stromal score was constructed, and the top 30 genes with connectivity were selected. Univariate Cox proportional hazard model analysis of overall survival (OS) of 161 GBM patients in the TCGA database was performed on co-up-regulated and down-regulated DEG between immune score and stromal score by using R4.1.0 software to obtain the DEG affecting OS. The intersection of the DEG obtained from PPI analysis and Cox analysis was taken as the prognostic core genes. According to the median expression value of prognostic core genes in GBM samples from the TCGA database, 161 patients were divided into prognostic core genes high expression group and low expression group (patients whose scores were equal to the median were not involved in the grouping), with 80 cases in each group. Kaplan-Meier survival analysis of OS was performed by using R4.1.0 software. GSEA 4.2.1 software was used to perform gene set enrichment analysis (GSEA) on all genes with transcriptome data of GBM patients in the two groups of the TCGA databases, and the main enriched functions of the two groups of genes were obtained. The CIBERSORT algorithm was used to test the accuracy of the proportion of tumor infiltrating immune cell (TIC) subsets in 169 GBM samples from the TCGA database, and 57 GBM samples were finally obtained. Immune cells with differential expression levels and immune cells related to the expression of prognostic core genes among the samples with different expression levels of prognostic core genes were analyzed; Venn program was used to obtain the intersection of immune cells with differential levels and related immune cells, and differentially expressed TIC related to expressions of prognostic core genes in GBM were obtained.Results:Based on the immune score and stromal score of GBM samples in the TCGA database, a total of 693 co-up-regulated and co-down-regulated DEG of both scores were screened out. After the intersection of 78 DEG related to OS obtained by univariate Cox regression analysis and 30 DEG obtained by PPI network results, CC motif chemokine receptor 2 (CCR2) was identified as the prognostic core gene ( HR = 1.294, 95% CI 1.060-1.579, P = 0.011). GBM patients with CCR2 high expression had worse OS compared with those with CCR2 low expression ( P = 0.009). GSEA analysis showed that genes in the CCR2 high expression group were mainly enriched in immune-related pathways, while genes in the CCR2 low expression group were mainly enriched in metabolism-related pathways. Among 57 screened GBM samples, there were differences in the levels of 3 immune cells between the CCR2 high expression group and the CCR2 low expression group ( P < 0.05). CCR2 expression was correlated with the levels of 9 immune cells (all P < 0.05). Venn program analysis showed that differentially expressed 3 TIC in GBM related to CCR2 gene expression were obtained; among them, M2 macrophages were positively correlated with CCR2 expression, while T follicular helper cell and activated NK cells were negatively correlated with CCR2 expression. Conclusions:CCR2 may be the core gene related to the prognosis in the TME of GBM. As reference, the level of CCR2 can help to predict the status of TME and prognosis in GBM patients, which is expected to provide a new direction for the treatment of GBM.
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ObjectiveTo investigate the distribution of vascular cognitive impairment （VCI） with kidney Yang deficiency syndrome and explore the biological nature of VCI with kidney Yang deficiency syndrome from the perspective of DNA methylation under the combination of disease and syndrome， so as to provide an epigenetic target for traditional Chinese medicine （TCM） treatment of this disease with this syndrome in the future. MethodCommunity residents in Beijing were screened out for cognitive impairment from September 2020 to November 2022 through the scale， and VCI patients were analyzed for the syndrome. VCI patients with kidney Yang deficiency syndrome and healthy people were enrolled in this study. Peripheral venous blood was collected and subjected to genome-wide DNA methylation detection by Illumina Human Methylation 850K BeadChip. Then， differentially methylated genes （DMGs） were screened out for bioinformatics analysis. ResultA total of 1 902 people were investigated in this study， and 201 of them had VCI， accounting for 10.57%， including 72.14% with kidney Yang deficiency syndrome. The methylation results showed that compared with the normal group， the VCI group had 386 differential methylation sites， and 136 DMGs were annotated. The Kyoto Encyclopedia of Gene and Genomes（KEGG） signaling pathway enrichment analysis showed that the DMGs between the two groups were mainly involved in mammalian target of rapamycin（mTOR） signaling pathway， Estrogen signaling pathway， cyclic adenosine monophosphate（cAMP） signaling pathway， etc. Protein-protein interaction （PPI） analysis showed that DMGs， such as epidermal growth factor receptor（EGFR）， epidermal growth factor （EGF）， and signal transducer and activator of transcription 3（STAT3）， played important roles in the network. ConclusionKidney Yang deficiency is the main syndrome in VCI patients. DMGs including EGFR， EGF， and STAT3 and the related pathways such as mTOR signaling pathway， Estrogen signaling pathway， and cAMP signaling pathway may play a vital role in the occurrence and development of VCI with kidney Yang deficiency syndrome.
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Objective@#To observe the refractive status of preschool children, and to explore the prevalence and development trend of ametropia in preschool children, so as to provide support for controlling adolescent ametropia and improving children s health care service.@*Methods@#This cross sectional survey was conducted among 27 561 preschool children (55 122 eyes) aged 3-6 years old in the High tech Zone of Suzhou from September to December 2020. The refractive status was screened by the US Weilun binocular optometry.@*Results@#The total detection rate of ametropia was 9.5%, including 8.6% of astigmatism, 1.3% of hyperopia and 0.5% of myopia. The detection rates of myopia (1.1%) and hyperopia (2.2%) were the highest in the 6-year-old group, and the older the age was, the lower the spherical diopter was, and the higher the cylindrical diopter was. The abnormal rate of colposcopy in girls ( 1.3% ) was higher than that in boys (0.9%), and the abnormal rate of total anisometropia in women (2.3%) was also higher than that in men (1.9%). The main astigmatism was mixed astigmatism (49.1%) and compound hyperopia astigmatism (39.2%); The older the age, the lower the detection rate of compound hyperopia astigmatism. And it is dominated by regular astigmatism( 97.5% ); The higher the age, the higher the detection rate of astigmatism with the rule, while the lower the detection rates of astigmatism against the rule and oblique axis astigmatism.@*Conclusion@#The detection rate of myopia and hyperopia increased significantly at the age of 6, and anisometropia and axial astigmatism also reached the highest at the age of 6. Local health care departments should pay attention to children s astigmatism, especially astigmatism with the rule.
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Glioma is the most common primary intracranial tumor. At present, the conventional treatment methods have limited effect and cannot significantly prolong the survival time of patients. Chemokine CCL2 is the most important member of the CC chemokine family, which can regulate glioma angiogenesis, immunosuppression, progression and invasion, and resistance to apoptosis. This article reviews the potential mechanism of CCL2 promoting the malignant progression of glioma, in order to provide new ideas and methods for the targeted therapy of glioma.
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Objective To investigate the antithrombotic function of traditional medicine salvia and aspirin in the myocardial infarction rats. Methods The myocardial infarction animal model was established by ligating left anterior descending coronary artery. The rats were randomly divided into the model group, the aspirin group, the salvia miltiorrhiza group, and the combination therapy group, with 8 rats in each group. The salvia miltiorrhiza group was injected with Danshen freeze-dried powder 17 mg/kg via tail vein injection, aspirin group was given aspirin 10 mg/kg, the combination group was given aspirin 10 mg/kg with the tail vein injected with Danshen freeze-dried powder 17 mg/kg, and the model group was given an equal volume of normal saline with an equal volume of saline injected into the tail vein. Continuous administration last 10 days, once daily. The blood coagulation, platelet aggregation, thromboxane B2, 6-Keto-F1α and von Willebrand factor were detected and compared. Results Compared with the model group, the maximum platelet aggregation rate(32.55％ ± 9.57 ％, 32.16％ ± 10.76％, 19.74％ ± 6.70％ vs. 58.75％ ± 4.81％) in the rats of the aspirin group, the salvia group and the combined treatment group significantly decreased, and the contents of TXB2 (70.58 ± 9.31 ng/ml, 73.10 ± 11.33 ng/ml, 49.25 ± 5.33 ng/ml vs. 107.86 ± 17.45 ng/ml) decreased (P<0.01). The maximum platelet aggregation rate and contents of TXB2 in the combination group were lower than those in the aspirin group and the salvia group (P<0.01). Compared with the model group, the 6-Keto-PGF1α content (67.64 ± 7.12 ng/ml, 81.72 ± 10.72 ng/ml vs. 57.80 ± 11.19 ng/ml) of the aspirin group and the combined treatment groupwas increased (P<0.05), prothrombin time (13.11 ± 0.67 s, 15.85 ± 0.25 s vs. 10.77 ± 0.46 s) prolonged (P<0.05), vWF (51.31 ± 4.12 ng/ml, 47.72 ± 10.32 ng/ml vs. 128.81 ± 11.14 ng/ml) decreased (P<0.05). The contents of 6-Keto-PGF1α of the combined treatment group was increased compared with aspirin group and salvia group (P<0.05), prothrombin time decreased than aspirin group (P<0.05) and Salvia group and vWF decreased than Salvia group (P<0.05). Conclusions The salvia miltiorrhiza and aspirin combination therapy has synergistic effect on anti-platelet aggregation and anticoagulant effect. The salvia miltiorrhiza and aspirin combination therapy showed small bleeding risk.
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Objective To systematically evaluate the diagnostic value of treadmill exercise test (TET) compared with dynamic electrocardiography (DCG). Methods Databases including Cochrane library, MEDLINE,EMbase,Google Scholar and CNKI were retrieved to collect randomized clinical trials (RCT) and controlled clinical trials(CCT)focusing on comparison of TET and DCG. After screening according to the inclusion and exclusion criteria,all articles received Jadad quality assessment,and were analyzed using Review Manager Version 5.3 software(The Cochrane Collaboration,Oxford,United Kingdom). Results 9 papers with a total of 667 patients who underwent coronary angiography(CAG),TET and DCG were eventually enrolled. Results of the present Meta?analysis were as below.(1)Sensitivity:9 trials(n = 552)suggested that the sensitivity is 80.62% in TET Group versus 72.64%in DCG Group[RR=1.11,95%CI(1.04~1.18),P=0.002]. The difference was statistically significant.(2)Specificity:7 trails(n=126)indicated that no statistical significance[RR=0.86,95%CI(0.71~1.04),P=0.11]was found in specificity when comparing TET(68/115)and DCG(87/126). Conclusion TET has higher sensitivity but similar specificity in comparison with DCG,and it turns out to be a simple and feasible inspecting method in diagnosing coronary heart disease.
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Objective To investigate the expression of IFN-γ, IL-5, IL-17 and TGF-βby Th cells and Th17 cells in rats with allergic rhinitis upon the intervention of IL-10.Methods SD rats were ran-domly divided into three groups including allergic rhinitis ( AR ) group, IL-10 treated group and control group (n=10).Rats in AR group and IL-10 treated group were sensitized by injection of ovalbumin (OVA) and aluminum hydroxide on the 1st, the 7th and the 14th days.The rats treated with equal volume of saline were set up as the control.The corresponding interventions ( OVA, OVA and IL-10, saline) were respec-tively given to rats in each group on the 21th day for 7 consecutive days.The clinical manifestations in rats were observed within 30 minutes after each administration.Serum samples were collected at 48 hours after the last challenge for the detection of IgE and OVA-sIgE.ELISA and Western blot assay were performed to detect IFN-γ, IL-5, IL-17 and TGF-βin nasal mucosa samples.Results Some characteristic symptoms of AR were observed in rats from AR group and IL-10 treated group.Compared with IL-10 treated rats, rats in AR group showed severe clinical symptoms such as constant rubbing and tearing of the eyes (P<0.05).The levels of IgE and OVA-sIgE in serum samples and the levels of IFN-γ, IL-5, IL-17 and TGF-βin nasal tis-sues were significantly increased in rats with RA (P<0.05), but were reduced with IL-10 intervention (P<0.05).Conclusion Exogenous IL-10 could be used to treat AR by reducing the expression of IFN-γ, IL-5, IL-17 and TGF-βin nasal tissues.
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Objective To observe the influence of p53-upregulated modulator of apoptosis (PUMA) on the growth of human brain glioma cell lines U251 (p53 mutant) and SHG-44 (p53 wild type),and to explore its possible mechanism.Methods Construct the adenovirus PUMA (Ad-PUMA) and vector of adenovirus (AdDsRed) which were respectively transfected into glioma cell lines U251 and SHG-44.Cells proliferation rates were measured with cell counting kit-8 (CCK-8).The apoptotic ratios were detected by flow cytometry.The expression of PUMA and apoptosis associated proteins (bcl-2,Bax) were determined with Western blot analysis.Caspase-3,Caspase-8,Caspase-9 activity were measured by Caspase activity assay kit.Results Compared with vector group and blank control group,Ad-PUMA transfected group showed strong cell proliferating inhibition effects [the inhibition rates were (50.89±4.73) ％ and (44.45±5.33) ％ respectively,P ＜0.05] and pro-apoptotic effects [apoptotic rates were (44.89±5.08) ％ and (31.67±7.32) ％,P ＜ 0.05] in different p53 glioma cell lines U251 and SHG-44.Western blot analysis showed that PUMA protein expression increased after Ad-PUMA transfection,accompanied by the reduced expression of the anti-apoptotic protein bcl-2 and the increased expression of pro-apoptotic protein Bax.The activity of Caspase testing results showed that the Caspase-3,Caspase-9 activity increased significantly,while the Caspase-8 activity changed little.Conclusion No matter how p53 phenotype,PUMA can inhibit glioma proliferation,promote apoptosis,and its mechanism may be through the mitochondrial apoptotic pathways,upregulation of Bax and inhibition of bcl-2 expression,which activated Caspase-9.Ad-PUMA is expected to become a new target for gene therapy of gliomas.
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Objective To explore differences of drug resistance of temozolomide(TMZ) on different CD133 immune prototype of glioma cells and study on the changes of their sensitivity to TMZ through increased PUMA. Methods CD133+-U87MG cells sorted by CD133 magnetic beads were cultured in serum-free stem cell medium respectively. The cells were infected with recombinant adenovirus, Ad-PUMA, diluted in cell culture medium with or without TMZ intervention.The inhibitory rate of cell proliferation was detected by MTT assay and 50 ％ inhibition concentration of TMZ was calculated. Apoptosis rates of CD133+-U87MG cells were assessed by flow cytometry (FCM) before and after intervention of exogenous PUMA and TMZ.Results The TMZ IC50 values of CD133+glioma cells were higher than that of CD133- glioma cells. There were significant differences in apoptosis rate between CD133+ glioma cell and CD133- glioma cell (all P＜0.05).Conclusion AdPUMA joint TMZ can promote glioma stem cells apoptosis, thus improve the sensitivity to chemotherapy of glioma.
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Objective To investigate the inhibitive effects of Ad-PUMA combined with temozolomide on human glioblastoma cells growth in vivo experiments. Methods The nude mouse model with human glioblastoma cells subcutaneous transplantation was established. The mice were randomly divided into 4 groups to receive subcutaneous injection at the 14th day separately with: Normal saline 100 μl (control, n=8), Ad-PUMA 2×108 pfu/100 μl (PUMA group, n=8), 10 mg/kg TMZ (TMZ group, n=8) and 2×108 pfu/100 μl Ad-PUMA + 10 mg/kg TMZ (combined group, n=8). Mice were killed after 20 days treatment.Tumor volume, inhibition rates and apoptotic index (AI) were measured, meanwhile, apoptotic tumor cells were detected by TUNEL technology respectively. The expression of MGMT mRNA and MGMT protein were revealed by the methods of RT-PCR and Western blot. Results According to the order: control group, AdPUMA group, TMZ group, combined group, tumor volumes were (3.68±0.09), (2.63±0.13), (2.13±0.07),(0.97±0.02) cm3 respectively (P＜0.05); the inhibitive rates were 0, 28.5 %, 42.1%, 73.6 % respectively and AI were (2.0±1.2) %, (11.4±2.6) %, (7.6±3.2) %, (20.6±8.6) % (P＜0.05). The results of Western blot and RT-PCR showed that MGMT mRNA and MGMT protein levels in TMZ group were higher than other groups (all P＜0.01). Conclusion Ad-PUMA combined with TMZ greatly enhances the sensitivity of human glioblastoma cells to TMZ and could effectively inhibit the proliferation and promote the apeptosis of glioblastoma cells, its mechanism was probably related Ad-PUMA promote apoptosis and inhibit MGMT expression.
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Objective To investigate the role of endoplasmic reticulum stress ( ERS) in human brain gliomas cell(SHG-44) apoptosis induced by proteasome inhibitor MG-132. Methods Human glioma cells were passage cultured. Glioma cells were treated by MG-132 with varying concentration(5, 10, 15 and 50 μmol/L) for 24 h. Compared with cells prior to the treatment (control group), cell viability was detected by MTT assay and the expression of ERS associated proteins GRP78 and apoptosis associated proteins Caspsse-12 was examined by PCR and Western-blotting. Results After MG-132 treatment for 24 h, SHG-44 cell viability was decreased significantly (39 %) (P <0.05), and continued to show a significant decline with the increasing concentration of MG-132 (P <0.05). RT-PCR results showed that the expression of ERS associated proteins GRP78 in SHG-44 cells were significantly increased after 5, 10, 15 and 50 μmol/L MG-132 treatment, and the expression of Caspase-12 was significantly increased after 5 μmol/L MG-132 treatment, slightly increased after 10 and 15 μmol/L treatment compared with that after 5 μmol/L treatment and reached the peak after 50 μmol/L treatment. Western-blotting results of GRP78 in SHG-44 cells were same as results of RT-PCR. Conclusion ERS may be involved in the apoptosis of gliomas cells induced by proteasome inhibitor MG-132.
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With the advent of the cancer stem cell hypothesis,the field of cancer research has experienced a revolution in how we think of and approach cancer. The discovery of "tumor-initiating cell" has offered an explanation for several long-standing conundrums on why tumors behave the way they do to treatment. Despite the great amount of research that has been done in order to understand the molecular aspects of tumors,the prognosis of tumors remains dismal. The slow progress in extending the survival of patients with malignant tumors is very likely due to poor understanding of the cell of origin in these tumors.This review article discusses the progress in our understanding of tumor-initiating cell as the cell of origin in cancers. We review the different proposed mechanisms of how tumor-initiating cell may originate,the molecular mechanisms of cancer initiation and progression, and finally the clinical implications of this research.
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Objective To explore the expression of CD133 and nestin in human glioma cell of patients with neuroglioeytoma and to see whether CD133 and nestin expression associate with the differentiation and pathologic grading.Methods The expression of CD133 and nesfin was detected by immunohistochemistry staining in 65 cases of human glioma and 19 specimens of normal brain tissues.Results The rate of the CD133 positive ceUs was 18.46%in human glioma and 0 in the control group(P＜0.01),and that of nesfin was 23.79%and 5.16%respectively.The positive cell rates of CD133 or nestin varied accordingly to different pathological grades.There were significant difference between grade Ⅰ and grade Ⅲ or grade Ⅳ (P＜0.01).There were difference between grade Ⅱ and grade Ⅲor grade Ⅳ (P＜0.01).The same significance were also found between grade Ⅲ and grade Ⅳ(P＜0.01).The higher pathological grades had higher positive cell rates.There was a significant corl~lation between the rate of CD133 positive cells and that of nestin in experimental group (r=0.408,P＜0.01).Conclusion Detecting CD133 and nestin in the human glioma can be used in diagnosing,judging the malignancy degree and the prognosis.
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OBJECTIVE: To investigate the situation and future trend of use of drugs in this hospital after medical re- form.METHODS:Using the principle of sequencing analysis of DUD system of drug,we analysed the data of drug consumption in our hospital during the period 1999-2000.RESULTS: The policy of medical reform has affected the component ratio of drugs used, the consumption of peroral drugs being increased.The incidence of cardiovascular diseases was high in this re- gion.CONCLUSION:Most of the drugs used after medical reform were those assigned in medical insurance,which will improve the rationality and effectiveness of medication.