Logic-gated tumor-microenvironment nanoamplifier enables targeted delivery of CRISPR/Cas9 for multimodal cancer therapy

Recent innovations in nanomaterials inspire abundant novel tumor-targeting CRISPR-based gene therapies. However, the therapeutic efficiency of traditional targeted nanotherapeutic strategies is limited by that the biomarkers vary in a spatiotemporal-dependent manner with tumor progression. Here, we propose a self-amplifying logic-gated gene editing strategy for gene/H2O2-mediated/starvation multimodal cancer therapy. In this approach, a hypoxia-degradable covalent-organic framework (COF) is synthesized to coat a-ZIF-8 in which glucose oxidase (GOx) and CRISPR system are packaged. To intensify intracellular redox dyshomeostasis, DNAzymes which can cleave catalase mRNA are loaded as well. When the nanosystem gets into the tumor, the weakly acidic and hypoxic microenvironment degrades the ZIF-8@COF to activate GOx, which amplifies intracellular H+ and hypoxia, accelerating the nanocarrier degradation to guarantee available CRISPR plasmid and GOx release in target cells. These tandem reactions deplete glucose and oxygen, leading to logic-gated-triggered gene editing as well as synergistic gene/H2O2-mediated/starvation therapy. Overall, this approach highlights the biocomputing-based CRISPR delivery and underscores the great potential of precise cancer therapy.

Circulation miR-193 cluster serves as a promising biomarker for diagnosis of breast cancer / 临床检验杂志

Objective@#To evaluate circulating miR-193 cluster as a biomarker for diagnosis of breast cancer (BC). @*Methods@#A total of 130 BC patients and 45 healthy controls (HCs) were enrolled. The level of miR-193 cluster was analyzed by using GEO database. RT-qPCR was used to evaluate the level of miR-193 cluster in serum of participants. Receivers operating characteristic (ROC) curve analysis was manipulated to investigate their diagnostic value for BC. @*Results@#In the screening stage, the results of GEO database indicated the expression of miR-193a-5p and miR-193b were decreased in the serum of BC patients with prior diagnostic value, and the significant decreased expressions of miR-193 cluster members, miR-193a-3p, -5p and miR-193b-3p, were observed in serum of BC patients compared with those in healthy controls by the determination of RT-qPCR（all P ＜0.01）. In the validation stage, the AUCROC of miR-193a-5p and miR-193b-5p were 0.888 (95% CI :0.808-0.969) and 0.954 (95% CI :0.902-1.000), respectively. In addition, the combined AUCROC of these two miRNAs was 0.982 (95% CI :0.966-0.999). @*Conclusion@#miR-193a-5p and miR-193b-5p in serum could be served as diagnostic biomarker for screening of breast cancer, and the combined detections of the two miRNAs may exhibit more diagnostic efficiency and good clinical application potential.

Overexpression of long noncoding RNA HMMR-AS1 promotes progression of lung adenocarcinoma / 临床检验杂志

Objective@#The purpose of this study is to explore the biological function of long non-coding RNA (lncRNA) HMMR-AS1 in proliferation and metastasis of lung adenocarcinoma (LUAD). @*Methods@#Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of HMMR-AS1 and its sense strand HMMR in LUAD cell lines. Then we knock down the HMMR-AS1 expression through small interfering RNA and evaluate the transfection efficiency and its effect on the expression of HMMR. CCK-8 (cell counting kit), clone formation, flow cytometric analysis, wound scratch assay and transwell assay were used to assess the biological function of A549 and H1299 cells. Western blot was used to detect the protein expression of HMMR in the two cell lines after transfection with si-HMMR-AS1. @*Results@#The expression of HMMR-AS1 in A549 and H1299 cells of LUAD cell line was markedly higher than that in normal lung epithelial cell BEAS-2A by upregulating approximately 3.06 and 5.02 folds (P＜0.05), respectively. After transfection with si-HMMR-AS1, the expression of HMMR-AS1 markedly reduced in both levels of transcription and protein (P＜0.05). Furthermore, knocking down of HMMR-AS1 significantly inhibited the proliferation, migration and invasion abilities, and increased the apoptosis rates of A549 and H1299 cells. @*Conclusion@#LncRNA HMMR-AS1 could promote malignant progression of LUAD cells through enhancing the growth, migration and invasion ability of LUAD cells.

Protective effects and its mechanism of rebamipide on aspirin-induced injury in human gastric mucosal epithelium cells / 中华消化杂志

Objective To investigate the protective effects and its mechanism of rebamipide on aspirin-induced injury in human gastric mucosal epithelium cells (GES-1).Methods GES-1 cells monolayer culture model was established in vitro.Then the cells were divided into negative control group,aspirin injured group and combination of rebamipide at different concentration (0.2,0.5,1.0 mrnol/L) and aspirin groups.The cell proliferation,the content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) of each group were detected.The ultrastructural changes of each group were observed by transmission electron microscopy (TEM).The expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) at protein level in the cells of each group were detected by Western blot.Nrf2 interfering suppression test was performed and then the influence of Nrf2 small interfering RNA (siRNA) on the expression of HO-1 protein was observed.One-way analysis of variance was performed for comparison among multi-groups and t-test was used for comparison between the two groups.Results The cell viability of aspirin injured group and combination of rebamipide at different concentration (0.2,0.5,1.0 mmol/L) and aspirin groups were (49.56±3.88)％,(59.34±4.36) ％,(70.79 ± 5.96) ％ and (86.07 ± 5.20) ％,respectively,and the difference was statistically significant (F=30.634,P＜ 0.01).Compared with aspirin injured group,the content of MDA significantly lowered in combination of rebamipide at different concentration (0.2,0.5,1.0 mmol/L) and aspirin groups ((2.26±0.25) nrnol/rng vs (1.85±0.13) nmol/mg vs (1.62±0.11) nmol/mg vs (1.13±0.15) nmol/mg),and the difference was statistically significant (F=23.821,P＜0.05).Compared with aspirin injured group,the activity of SOD significantly increased in combination of rebamipide at 0.5 and 1.0 mmol/L and aspirin groups ((8.49±0.89) U/rng vs (11.50±1.03) U/mg vs (13.74±0.76) U/mg),the difference was statistically significant (F=25.666,P＜0.05).Under TEM,the cell ultrastrucmral was obviously inured in aspirin treated,while rebamipide could relieve the injury.The differences of relative expression quantity of Nrf2 and HO-1 at protein level among combination of rebamipide at 0.2,0.5 and 1.0 mmol/L and aspirin groups and aspirin injured group were statistically significant (0.35±0.04 vs 0.46± 0.05 vs 0.84±0.08 vs 0.15±0.02,0.72±0.09 vs 0.93±0.11 vs 1.29±0.14 vs 0.39±0.07,F=92.550and 38.235,both P＜0.05).After transfected with Nrf2 siRNA,the expression of HO-1 was 0.38±0.04 in aspirin injured group and 0.62±0.08 in combination of rebamipide and aspirin group,which was lower than that before transfection (0.61 ± 0.05,1.33± 0.09),respectively.The differences were statistically significant (t =6.276 and 10.444,both P＜0.05).Conclusion Rebamipide may activate Nrf2/HO-1 pathway and relieve aspiriwinduced oxidative stress in GF1 ceils.

Expression of miR-224 in diffuse large B cell lymphoma and its clinical significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of miR-224 in diffuse large B cell lymphoma (DLBCL) and its relationship with clinical pathological features and prognosis.</p><p><b>METHODS</b>Real-time PCR was used to detect the expression of miR-224 in 168 DLBCL and 25 normal lymphoid tissues.</p><p><b>RESULTS</b>The expression of miR-224 in DLBCL (0.97 ± 0.33) was significantly lower than that in normal lymphoid tissues (1.87 ± 0.43, P<0.05). There were no significant correlations between the miR-224 expression and age (P=0.434), gender (P=0.613) tumors stage (P=0.250), IPI (P=0.355) and lactate dehydrogenase (P=0.398). Using the median of miRNA-224 expression as threshold, we subdivided patients into low and high expression group. The five-year progression-free survival and overall survival were significantly lower in low expression group as compared to those in high expression group.</p><p><b>CONCLUSION</b>miR-224 expression may play an important role in the development and progression of DLBCL and could be prognostic significance.</p>

Expression of E-cadherin in the tumor tissue and serum of patients with esophageal squamous cell carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To survey E-cadherin (E-cad) expression in tumor tissue and serum of esophageal squamous cell carcinoma patients, and to observe the clinical significance of their expression.@*METHODS@#Forty-eight samples of esophageal squamous cell carcinoma tissue, 23 samples of erosive esophagitis tissue, 24 samples of normal esophagus tissue and the corresponding sera were obtained. We used immunohistochemistry (IHC) to detect expression of E-cad in the tissues and enzyme-linked immunosorbent assay (ELISA) to examine expression of E-cad in the serum. Furthermore, we collected complete clinicopathological data from the participating patients.@*RESULTS@#The expression level of E-cad in the esophageal squamous cell carcinoma tissue was lower than that in normal esophagus tissues and erosive esophagitis tissues (P<0.05). Moreover, the expression level of E-cad was related to the depth of invasion, the status of lymph node metastasis and the level of differentiation of esophageal squamous cell carcinoma (P<0.05). The expression level of serum E-cad of esophageal squamous cell carcinoma patients was obviously higher than that in the serum of normal esophagus controls and erosive esophagitis patients (P<0.05). But the expression level of E-Cad in the serum of esophageal squamous cell carcinoma patients was unrelated to clinicopathological features. The expression level of E-cad in the tissue was not correlated with that in the serum(P=0.134).@*CONCLUSION@#The expression of E-cad in tissues may assistin the diagnosis and prognosis of esophageal squamous cell carcinoma. The expression of E-cad in the serum may assistin the diagnostic screening of esophageal squamous cell carcinoma.

The study on the mechanim of clopidogrel in human gastric epithelial GES-1 cell line injury / 中华消化杂志

ObjectiveTo explore the mechanism of clopidogrel in human gastric epithelial cell line (GES-1) injury.MethodsSet up GES-1 cells monolayer culture model.Then the GES-1 cells were divided into negative control group,U0126 intervented group,clopidogrel intervented group and combined intervented group (U0t26 treated firstly then clopidogrel intervented).The cell proliferation and apoptosis in each group was examined by methyl thiazolyl tetrazolium (MTT) assay and Flow cytometry.TheexpressionofphosphorylatedERK1/2ineachgroupwasdetectedby immunocytochemistry method,and the expression quantity of phosphorylated ERK1/2 in each group was measured by western blot.ResultsThe result of MTT assay showed that compared with negative control group,the proliferation of GES-1 cells was inhibited in U0126 group,clopidogrel group and combined intervented group,and the inhibition percentage was 21.8％ ±2.7％,46.3％ ± 3.4％ and 82.9 ％ ± 0.8 ％ respectively ( F=615.556,P =0.000 ).The result of immunocytochemistry indicated that the expression of p-ERK in U0126 group,Clopidogrel group and combined intervented group decreased compared with negative control group,which was 10.80±1.64,7.20± 1.64,4.40±0.89and 1.40±0.55 respecitively (F=49.426,P=0.000).The result of western blot and immunocytochemistry was of the same trend.Conclusion In GES-1 cell model,clopidogrel may injureGES-1 cells through MAPK/EPK signal transduction pathway.

Expressions of interferon-inducible genes in patients with systemic lupus erythematosus and their assoc-iation with disease activity / 中华风湿病学杂志

Objective To investigate the expression levels of interferon-inducible genes (IFIT1,IFIT4,OAS1,OASL,ISG15) in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus(SLE).and the relations between these genes expression levels and disease activity are explored.Methods Sybr green dye based real-time quantitative PCR method was used to detect the expression levels (indicated as-△△Ct value) of WIT1,IFIT4.OAS1,OASL and ISG15 in 76 patients with SJJE and 54 controls.Their expression levels were compared with erythroeyte sedimentation rate (ESR),serum C reactive protein (CRP),complement C3,C4.antinuclear antibody (ANA).anti-double stranded DNA antibody.The associations between the expression levels of IFIT1,IFIT4,OASI.OASL,ISG15,ESR,CRP,complement C3,C4,ANA,anti-double stranded DNA antibody and SLEDAI scores in patients with SLE were analyzed.Results ① The expression levels of WIT1,IFIT4,OAS1,OASL and ISG15 in the SLE patients were significantly higher than those of the normal controls (P＜0.01).The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in active SLE patients were higher than those of inactive SLE patients (P＜0.05).The real time expression levels of IFIT1,IFIT4,OAS1.OASL and ISG15 showed positive correlations with each other (r＞0.5,P＜0.05) in patients with SLE.② The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 were positively correlated with the SLEDAI scores (r＞0.5,P＜0.05).③ There was no correlation between ESR,CRP,complement C3,C4,ANA and the expression levels of IFIT1,IFIT4,OAS1,OASL,ISG15,SLEDAI scores except anti-double stranded DNA antibody (r＞0.5.P＜0.05).Conclusion The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in patients with SLE are significantly higher than those of the normal controls,and positively associated with SLEDAI scores,so they are helpful in evaluating SLE disease activity and severity.IFIT1,IFIT4,OAS1,OASL and ISG15 genes may be the potential treating targets for SLE.

High dose epirubicin containing combination regimen in the treatment of advanced breast cancer / 中国癌症杂志

Purpose:To study the efficacy and the toxic side effects of high dose epirubicin containing combination regimen in the treatment of advanced breast cancer. Methods:16 patients with advanced breast cancer were treated with epirubicin (EPI) 100 mg/m 2,CTX 600 mg/m 2,5 FU 500 mg/m 2,every 21 days,Each patient was given at least 2 cycles. Results:There were 2 CR and 9 PR, the overall response rate was 68.8% (11/16). Main side effects were grade Ⅰ—Ⅱ leucopenia, nausea and vomitting, alopecia.Conclusions:This study confirmed that combination chemotherapy with high dose epirubicin is a safe and effective regimen for patients with advanced breast cancer and is worthy of further clinical trial.