Background and purpose:As the development of the completion of the human genome project (HGP), the research focus is turning to the gene function research. At present, the domestic experimental research on the apoptosis of K562 cells induced by antisense olignonucleotides is rare. This study was aimed to investigate the effect of human chronic myelogenou leukemia (CML) bcr-abl fusion gene antisense oligonucletides on autophagy and apoptosis of CMLK562 cells in vitro. Methods:By liposome as the carrier, K562 cells were transfected with the bcr-abl gene antisense olignonucleotides. Hoechst staining method was used to observe the apoptosis inducing effect of different concentrations of oligonucleotides, the expressions of LC3-Ⅱ, autophagy-related protein, were determined by the Western blot method, the cell cycles were determined by lfow cytometry (FCM), and JEM-4000EX electron microscope technology was used to detect the apoptosis morphological changes. The apoptosis was detected by DNA agarose gel electrophoresis. Results:Hoechst staining results showed that the bcr-abl gene antisense oligonucletides signiifcantly promoted the apoptosis of K562 cells in a certain concentration dependent manner. Western blot showed that the expression level of LC3-Ⅱwas obviously higher in bcr-abl gene antisense oligonucletides transfected group than the control group, showing a promoting effect on cell autophagy. FCM test results showed that bcr-abl gene antisense oligonucleotides transfected K562 cells showed obvious cell cycle arrest, visible obvious apoptosis morphology under the electron microscope, and DNA Ladder showed obvious apoptosis fragments. Conclusion:The bcr-abl gene antisense olignonucleotides can signiifcantly induce the cell apoptosis of K562. This study provides a new method for CML therapy.

Objective:To screen and characterize aptamers against BCR-ABL fusion protein.Methods:A 90bp single stranded DNA( ssDNA) random library was subjected to 13 rounds of selection against BCR-ABL fusion protein by systematic evolution of ligands by expotential enrichment ( SELEX ) method, the selected aptamers were cloned and sequenced.The primary sequences and structure of aptamers were analyzed by Clustal W and DNA Folding Sever and the percentage of the ssDNA pool bound to BCR-ABL core protein were determinated.Results: after 13 rounds selection, the percentage of ssDNA pool bound to BCR-ABL fusion protein increased from 0.3%to 47.1%,the results showed that affinities of the Aptamers were different,the second structure analysis revealed possible stem-loops for binding to BCR-ABL fusion protein,the affinity of aptamer A2 to BCR-ABL fusion protein was highest with Kd values as low as 72 nmol/L.Conclusion:Aptamers against BCR-ABL fusion protein has been identified by SELEX methods from a 90 bp single stranded DNA library.And provide certain reference for the clinical treatment of chronic myelogenous.

Objective To observe the effects of live combined bifidobacterium,lactobacillus and enterococcus powder on immunoglobulin E (IgE) and interleukin-17 (IL-17) in atopic children with bronchiolitis.Methods Sixty cases of atopic children with bronchiolitis were randomly divided into the therapy group (30 cases) and the control group (30 cases).Twenty-five healthy children were enrolled as the healthy control group.Both the therapy group and the control group were given traditional therapy.The therapy group received live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months.The change of IgE and IL-17 levels were observed during the acutestage,remission stage and after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2months.Results (1) The levels of IgE and IL-17 of therapy group[(132.36 ±9.50) μg/L and (77.76 ±7.95)μg/L] during acute stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ±4.79) μg/L] (all P ＜0.001).The levels of IgE and IL-17 of control group [(128.83 ± 8.06) μg/L and (76.61 ±6.18) μg/L] during remission stage were markedly higher than those in the healthy control group [(52.80 ±4.92) μg/L and (46.92 ± 4.79) μg/L] (all P ＜ 0.001).(2) The levels of IgE of therapy group (56.67 ± 9.20)μg/L after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than those in the control group (70.50 ± 11.38) μg/L (P ＜ 0.001).The levels of IL-17 of therapy group [(49.63 ± 6.35) μg/L] at the time after receiving live combined bifidobacterium,lactobacillus and enterococcus powder for 2 months were markedly lower than these in the control group (54.77 ± 6.33) μg/L (P =0.003).Conclusion Receiving live combined bifidobacterium,lactobacillus and enterococcus powder for two months can decrease the IgE and IL-17 levels in atopic children with bronchiolitis.

Objective To investigate the clinical significance and changes of serum interleukin-4 (IL-4),interferon-γ(IFN-γ) and immunoglobulin E (IgE) levels in infants with spasmodic laryngitis.Methods Serum samples were obtained from 50 children with spasmodic laryngitis and 30 healthy children.The changes of IL-4,IFN-γand IgE levels in serum when hospital admission (acute stage),before discharge (the stage of clinical symptom disappeared) and after 2 months of follow-up were observed.Enzyme-linked immunosorbent assay was used to determine the levels of IL-4 and IFN-γ in serum.Serum IgE level was determined with enzyme-linked fluoroimmuneassay.Results The serum level of IL-4 in patients with spasmodic laryngitis during acute stage were much higher than that in normal control group[(20.65 ±5.87) ng/L vs(8.23 ±2.71) ng/L,t =5.536,P ＜0.05].It was decreased during the stage of clinical symptom disappeared and significantly lower than that during acute stage[(11.07 ± 2.93) ng/L vs (20.65 ± 5.87) ng/L,F =5.258,P ＜ 0.05].The serum level of IFN-γ in patients with spasmodic laryngitis during acute stage were much lower than that in normal control group[(61.38 ±6.36) ng/L vs (108.42 ± 13.56) ng/L,t =4.831,P ＜ 0.05],and it was increased during the stage of clinical symptom disappeared [(95.12 ± 11.63) ng/L] and significantly higher than that during acute stage(F =4.239,P ＜ 0.05).There was no difference between the patients at clinical symptom disappeared stage and normal control group (P ＞ 0.05).Two months after discharge,there was no difference in serum INF-γlevel between the patients [(75.68 ±6.29) ng/L] and that in control group (P ＞ 0.05).The serum level of IgE in patients with spasmodic laryngitis during acute stage were much higher than that in normal control group [(136.63 ± 17.86) ng/L vs (47.16 ± 6.83) ng/L,t =6.685,P ＜ 0.05].It was decreased during the stage of clinical symptom disappeared [(88.07 ± 11.83) μg/L] and significantly lower than that during acute stage(F =5.182,P ＜ 0.05),but higher than that of normal control group (P ＜ 0.05).Two months after discharge [(86.72 ± 7.25) μg/L] it was still higher than that in control group (P ＜ 0.05).Conclusions There is TH1/TH2 imbalance in infants with spasmodic laryngitis.Maybe spasmodic laryngitis is one of the clinical manifestations of atopic diseases.IL-4,IFN-γand IgE may play important roles in immunologic pathogenesis.

Objective:To understand the influencing factors of death of epidemic Japanese encephalitis (EJE) cases in Longnan City of Gansu Province.Methods:In the EJE Monitoring Information Report Management System of the Chinese Disease Prevention and Control Information System, data on EJE cases with onset from 2014 to 2018 and current address in Longnan City were derived. An "Individual Questionnaire of Epidemic Japanese Encephalitis in Longnan City" was designed, retrospective study was conducted on enrolled cases, their information on demographic data, consultation, onset, clinical classification, and chronic underlying diseases were collected, characteristics of EJE cases and death-related factors were analyzed.Results:From 2014 to 2018, a total of 260 EJE cases were reported in Longnan City, and 259 cases completed the questionnaire. Among them, 70 cases (27.0%) were aged ≥60 years old, 67 cases (25.9%) were severe and extremely severe, and 55 cases (21.2%) had chronic underlying diseases. Among 259 EJE cases, 46 cases died, with a fatality rate of 17.8%. After multivariate unconditional logistic regression analysis, age ≥60 years old [odds ratio ( OR)=2.667, 95% confidence interval ( CI): 1.140-6.237], severe and extremely severe ( OR = 2.762, 61.820, 95% CI: 1.053-7.091, 5.149-742.239), and chronic underlying diseases ( OR = 2.489, 95% CI: 1.038-5.964) were risk factors for death in EJE cases. Conclusions:The influencing factors of death of EJE cases in Longnan City are age, clinical classification and chronic underlying diseases. Therefore, we should focus on patients over 60 years old, clinically classified as severe or extremely severe, and suffering from chronic underlying diseases, and strengthen the immunization of EJE vaccine for key populations.

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.

BACKGROUND: We have recently developed a new Coronary Artery Tree description and Lesion EvaluaTion (CatLet) angiographic scoring system. Our preliminary studies have demonstrated its superiority over the the Synergy between percutaneous coronary intervention (PCI) with Taxus and Cardiac Surgery (SYNTAX) score with respect to outcome predictions for acute myocardial infarction (AMI) patients. The current study hypothesized that the residual CatLet (rCatLet) score predicts clinical outcomes for AMI patients and that a combination with the three clinical variables (CVs)-age, creatinine, and ejection fraction, will enhance its predicting values. METHODS: The rCatLet score was calculated retrospectively in 308 consecutively enrolled patients with AMI. Primary endpoint, major adverse cardiac or cerebrovascular events (MACCE) including all-cause mortality, non-fatal AMI, transient ischemic attack/stroke, and ischemia-driven repeat revascularization, was stratified according to rCatLet score tertiles: rCatLet_low ≤3, rCatLet_mid 4-11, and rCatLet_top ≥12, respectively. Cross-validation confirmed a reasonably good agreement between the observed and predicted risks. RESULTS: Of 308 patients analyzed, the rates of MACCE, all-cause death, and cardiac death were 20.8%, 18.2%, and 15.3%, respectively. Kaplan-Meier curves for all endpoints showed increasing outcome events with the increasing tertiles of the rCatLet score, with P values <0.001 on trend test. For MACCE, all-cause death, and cardiac death, the area under the curves (AUCs) of the rCatLet score were 0.70 (95% confidence intervals [CI]: 0.63-0.78), 0.69 (95% CI: 0.61-0.77), and 0.71 (95% CI: 0.63-0.79), respectively; the AUCs of the CVs-adjusted rCatLet score models were 0.83 (95% CI: 0.78-0.89), 0.87 (95% CI: 0.82-0.92), and 0.89 (95% CI: 0.84-0.94), respectively. The performance of CVs-adjusted rCatLet score was significantly better than the stand-alone rCatLet score in terms of outcome predictions. CONCLUSION: The rCatLet score has a predicting value for clinical outcomes for AMI patients and the incorporation of the three CVs into the rCatLet score will enhance its predicting ability. TRIAL REGISTRATION http://www.chictr.org.cn , ChiCTR-POC-17013536.
Humans ; Coronary Artery Disease/complications* ; Death ; Myocardial Infarction/etiology* ; Percutaneous Coronary Intervention ; Retrospective Studies ; Risk Assessment ; Risk Factors ; Treatment Outcome

Osteoporotic vertebral compression fracture (OVCF) can lead to lower back pain and may be even accompanied by scoliosis, neurological dysfunction and other complications, which will affect the daily activities and life quality of patients. Vertebral augmentation is an effective treatment method for OVCF, but it cannot correct unbalance of bone metabolism or improve the osteoporotic status, causing complications like lower back pain, limited spinal activities and vertebral refracture. The post-operative systematic and standardized rehabilitation treatments can improve curative effect and therapeutic efficacy of anti-osteoporosis, reduce risk of vertebral refracture, increase patient compliance and improve quality of life. Since there still lack relevant clinical treatment guidelines for postoperative rehabilitation treatments following vertebral augmentation for OVCF, the current treatments are varied with uneven therapeutic effect. In order to standardize the postoperative rehabilitation treatment, the Spine Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized relevant experts to refer to relevant literature and develop the "Guideline for postoperative rehabilitation treatment following vertebral augmentation for osteoporotic vertebral compression fracture (2022 version)" based on the clinical guidelines published by the American Academy of Orthopedic Surgeons (AAOS) as well as on the principles of scientificity, practicality and advancement. The guideline provided evidence-based recommendations on 10 important issues related to postoperative rehabilitation treatments of OVCF.

Tumor cells along with a small proportion of cancer stem cells exist in a stromal microenvironment consisting of vasculature, cancer-associated fibroblasts, immune cells and extracellular components. Recent epidemiological and clinical studies strongly support that vitamin D supplementation is associated with reduced cancer risk and favorable prognosis. Experimental results suggest that vitamin D not only suppresses cancer cells, but also regulates tumor microenvironment to facilitate tumor repression. In this review, we have outlined the current knowledge on epidemiological studies and clinical trials of vitamin D. Notably, we summarized and discussed the anticancer action of vitamin D in cancer cells, cancer stem cells and stroma cells in tumor microenvironment, providing a better understanding of the role of vitamin D in cancer. We presently re-propose vitamin D to be a novel and economical anticancer agent.

Neoadjuvant chemotherapy has become an indispensable weapon against high-risk resectable cancers, which benefits from tumor downstaging. However, the utility of chemotherapeutics alone as a neoadjuvant agent is incapable of generating durable therapeutic benefits to prevent postsurgical tumor metastasis and recurrence. Herein, a tactical nanomissile (TALE), equipped with a guidance system (PD-L1 monoclonal antibody), ammunition (mitoxantrone, Mit), and projectile bodies (tertiary amines modified azobenzene derivatives), is designed as a neoadjuvant chemo-immunotherapy setting, which aims at targeting tumor cells, and fast-releasing Mit owing to the intracellular azoreductase, thereby inducing immunogenic tumor cells death, and forming an in situ tumor vaccine containing damage-associated molecular patterns and multiple tumor antigen epitopes to mobilize the immune system. The formed in situ tumor vaccine can recruit and activate antigen-presenting cells, and ultimately increase the infiltration of CD8⁺ T cells while reversing the immunosuppression microenvironment. Moreover, this approach provokes a robust systemic immune response and immunological memory, as evidenced by preventing 83.3% of mice from postsurgical metastasis or recurrence in the B16-F10 tumor mouse model. Collectively, our results highlight the potential of TALE as a neoadjuvant chemo-immunotherapy paradigm that can not only debulk tumors but generate a long-term immunosurveillance to maximize the durable benefits of neoadjuvant chemotherapy.