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.

Objective To study the effect and pathological mechanisms of the neuro-immune response to viral encephalomyelitis caused by virulence reversion following the intrathalamic neurovirulence test for poliomyelitis vaccine in rhesus macaques.Methods Stock solutions(≥7000 lgCCID50/L)of inactivated polio vaccines(Vero cells)of type I,type Ⅱ,and type Ⅲ Sabin strains and 10-1 dilution of each type of polio vaccine were given to macaques,which were subjected to a intrathalamic neurovirulence test.Using immunohistochemical method,the pathological changes caused by polio,as determined by the distribution of CD 155 and CD4+receptor T lymphocytes,CD20+B lymphocytes,and CD68+microglia,were detected.Results Lesions were observed on the virulence-reverted polio cases.Inflammatory cell infiltration,neuronal degeneration and necrosis,satellite phenomena,perivascular cuffing,and glial cell proliferation were observed in the spinal cord.The inflammatory cells in the perivascular cuffing and proliferative glial nodules were mainly CD4+T lymphocytes,CD20+B lymphocytes,and CD68+microglia.There was no significant difference in the distribution of the poliovirus receptor CD 155 in the neurons and glial cells of monkeys with and without polio,and no expression was observed in their vascular endothelial cells.Conclusions Polio caused by virulence reversion of the intrathalamic neurovirulence test is viral encephalomyelitis.

Objective: To investigate the expression of mannose binding lectin 2 (MBL2) in hepatocellular carcinoma(HCC) cell lines and its effects on the proliferation and prognosis. Methods: MBL2 expression levels in patients of different sexes and survival analysis of patients were subsequently performed using bioinformatics method. The mRNA and protein expression levels were detected with the qRT⁃PCR and Western blot. Cell proliferation was examined by MTT assay and cell clone formation assay;Flow cytometry was used to detect the cell apoptosis and cell cycle. Gene set enrichment analysis(GSEA) was used to analyze the signal pathways of MBL2 enrichment. Results: Bioinformatics analysis and qRT⁃PCR results showed that MBL2 was highly expressed in male HCC patients compared with female HCC patients , and was related to the prognosis of HCC patients. Compared with the control group ,over⁃expression of MBL2 inhibited the viability and clone formation rate of Huh7 cells and MHCC⁃97H cells;In addition , cell cycles were arrested and cell apoptosis increased. GSEA and Western blot results showed that MBL2 was enriched in the cell cycle signal pathway. Overexpression of MBL2 inhibited the expression of CDK4 protein and promoted the expression of P16 and BAX protein. Conclusion The expression of MBL2 is down regulated in HCC , which is related to the prognosis of male patients with HCC and participates in the process of cell proliferation. MBL2 may be a potential therapeutic target for male patients with HCC.

Animals ; SARS-CoV-2 ; Antibodies, Neutralizing ; Macaca mulatta ; COVID-19/prevention & control* ; Antibodies, Viral ; Vaccines

Objective:To investigate the hepatitis B surface antigen (HBsAg) clearance condition and its predictive factors after treatment with nucleos(t)ide analogues to pegylated interferon-α add-on therapy in patients with chronic hepatitis B.Methods:Patients with chronic hepatitis B who visited the First Affiliated Hospital of Zhengzhou University from 2018~2019 were prospectively enrolled. HBsAg≤ 1500 IU/mL, hepatitis B e antigen-negative, HBV DNA undetectable, received antiviral treatment with nucleos(t)ide analogues for at least one year, and pegylated interferon-α add-on therapy for 48 weeks were included. The primary endpoint of study was to determine the proportion of HBsAg clearance at 72 weeks. Concurrently, the predictive factors for HBsAg clearance were analyzed. Quantitative and qualitative data were analyzed using a t-test or non-parametric test and a Fisher's exact test.Results:A total of 38 cases were included in this study, of which 13 cases obtained HBsAg clearance at 48 weeks of therapy and another six cases obtained HBsAg clearance throughout the extended treatment period of 72 weeks, accounting for 50.00% of all enrolled patients. There was a significant difference in HBsAg dynamics between the HBsAg clearance group and the non-clearance group (P < 0.05). Univariate logistic regression analysis showed that patients' age, baseline, 12-and 24-week HBsAg levels, and early HBsAg reduction were predictive factors for HBsAg clearance at 72 weeks of treatment. Multivariate logistic regression analysis showed that age (OR = 1.311; P = 0.016; 95% confidence interval: 1.051~1.635) and HBsAg levels at 24 weeks of treatment (OR = 4.481; P = 0.004; 95% confidence interval: 1.634~12.290) were independent predictors for HBsAg clearance.Conclusion:Hepatitis B e antigen-negative, nucleos(t)ide analogue treated, HBsAg ≤ 1500 IU/mL, and HBV DNA undetectable, peg-IFNα add-on treatment for 48 weeks could promote HBsAg clearance in patients with chronic hepatitis B. Six of the sixteen cases (37.50%) who did not obtain HBsAg clearance at week 48 did so with the course of therapy extended to week 72. Hence, the optimal individualized treatment strategy should be customized according to the predictors rather than the fixed 48-week course. Age (≤ 38), baseline HBsAg level (≤2.86 log 10IU/ml), HBsAg level at 24 weeks (≤ 0.92 log 10IU/ml), and 12-week HBsAg decrease from baseline (≥ 0.67 log 10IU/ml) indicate that patients are highly likely to obtain HBsAg clearance at the 72 weeks of combination therapy, in which the combined indicator based on HBsAg level ≤0.92 log 10IU/ml at 24 weeks will identify 85.0% to 100.0% of patients with HBsAg clearance.

Objective : To detect the changes of autophagic flux at different stages after oxygen⁃glucose deprivationreoxygenation (OGD/R) with several highly sensitive methods. Methods : Primary cortical neurons after oxygen deprivation of sugar after reoxygenation (OGD/R) were divided into the experimental OGD/R group and OGD/R + bafilomycinA1 (BafA1) group , using an RFP⁃GFP series fluorescent tags LC3 gene transfection detection cytolysosome and fusion of lysosomes , transmission electron microscope (TEM) observation the ultrastructure of autophagy , p62/SQSTM1 combining LC3 protein to flip the experimental testing p62 and LC3 protein quantitative , p62 immune staining observing the distribution and content. Results : Under fluorescence microscope , the ratio of autophagy lysosome to autophagosome increased significantly in OGD/R group , and the changes of autophagy structure in different stages could be observed in TEM. The ratio of soluble p62 and LC3 Ⅱ/ Ⅰ reflected the activation of autophagic flux , and p62 was mainly distributed in BafA1 group after fluorescence staining. Conclusion Each method has its own advantages , and different methods and indicators can accurately reflect the specific changes of autophagic flux in different stages after neuronal OGD/R. Mastering and applying these methods can effectively explore central nervous system diseases from the perspective of autophagy.

Allografts ; COVID-19 ; China/epidemiology* ; Disease Outbreaks ; Humans ; Kidney Transplantation/adverse effects* ; SARS-CoV-2

Objective:To investigate the effect of Chinese reading aloud training (c-RAT) on cognitive function and activities of daily living in patients with vascular cognitive impairment no dementia (VCIND).Methods:From January 2018 to January 2019, totally 80 patients with VCIND in the General Hospital of Xingtai Mining Group were enrolled.Subjects were randomized grouped into c-RAT group ( n=43) and control group ( n=37). The c-RAT group was asked to read aloud the Chinese paper for 5 days a week for 12 weeks.At the time of enrollment and 12 weeks after the intervention, the Montreal cognitive assessment (MoCA), trail making test (TMT), auditory verb learn test (AVLT), digit symbol substitution test (DSST) and modified Barthel index (MBI) were filled out. Results:Compared with control group, the difference of D-value in c-RAT group was statistically significant in MoCA total scores(4.00(2.00), 1.50(0.50), Z=3.012, P=0.003), scores of MoCA-Visual space execution (2.00(1.00), 0.00(2.00), Z=2.787, P=0.008), MoCA-attention (1.00(1.00), 0.00(0.50), Z=2.369, P=0.022), MoCA-language (1.00(1.00), 0.00(0.75), Z=3.049, P=0.000)and MoCA-delayed recall(2.00(2.00), 0.00(1.00), Z=2.043, P=0.014), TMT-A scores (-8.00(23.00), 10.50(30.25), Z=2.120, P=0.039), AVLT scores (1earning)(3.00(2.00), 0.50(0.75), Z=2.266, P=0.039) , AVLT scores (recall)(2.00(1.00), 0.00(1.00), Z=2.974, P=0.003)、AVLT scores (recognition) (2.00(0.00), 0.50(1.50), Z=3.054, P=0.000)and DSST scores ((4.96±0.71), (2.39±0.78), t=2.572, P=0.014), while there were no significant differences in the rest parts of MoCA, TMT-B and MBI( P>0.05). The increased scores in DSST were positively correlated ( r=0.205, P=0.006) with the number of reading aloud tasks finished. Conclusion:C-RAT can improve general cognition, especially in information processing speed, executive function, attention and auditory memory.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Animals ; Brain ; physiology ; Consciousness ; physiology ; Humans ; Mice ; Neurotransmitter Agents ; physiology ; Optics and Photonics ; Synaptic Transmission ; physiology