An adeno-associated virus variant enabling efficient ocular-directed gene delivery across species.

Recombinant adeno-associated viruses (rAAVs) have emerged as promising gene therapy vectors due to their proven efficacy and safety in clinical applications. In non-human primates (NHPs), rAAVs are administered via suprachoroidal injection at a higher dose. However, high doses of rAAVs tend to increase additional safety risks. Here, we present a novel AAV capsid (AAVv128), which exhibits significantly enhanced transduction efficiency for photoreceptors and retinal pigment epithelial (RPE) cells, along with a broader distribution across the layers of retinal tissues in different animal models (mice, rabbits, and NHPs) following intraocular injection. Notably, the suprachoroidal delivery of AAVv128-anti-VEGF vector completely suppresses the Grade IV lesions in a laser-induced choroidal neovascularization (CNV) NHP model for neovascular age-related macular degeneration (nAMD). Furthermore, cryo-EM analysis at 2.1 Å resolution reveals that the critical residues of AAVv128 exhibit a more robust advantage in AAV binding, the nuclear uptake and endosome escaping. Collectively, our findings highlight the potential of AAVv128 as a next generation ocular gene therapy vector, particularly using the suprachoroidal delivery route.

Development and validation of a prediction model for early screening of people at high risk for colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is a serious threat worldwide. Although early screening is suggested to be the most effective method to prevent and control CRC, the current situation of early screening for CRC is still not optimistic. In China, the incidence of CRC in the Yangtze River Delta region is increasing dramatically, but few studies have been conducted. Therefore, it is necessary to develop a simple and efficient early screening model for CRC. AIM: To develop and validate an early-screening nomogram model to identify individuals at high risk of CRC. METHODS: Data of 64448 participants obtained from Ningbo Hospital, China between 2014 and 2017 were retrospectively analyzed. The cohort comprised 64448 individuals, of which, 530 were excluded due to missing or incorrect data. Of 63918, 7607 (11.9%) individuals were considered to be high risk for CRC, and 56311 (88.1%) were not. The participants were randomly allocated to a training set (44743) or validation set (19175). The discriminatory ability, predictive accuracy, and clinical utility of the model were evaluated by constructing and analyzing receiver operating characteristic (ROC) curves and calibration curves and by decision curve analysis. Finally, the model was validated internally using a bootstrap resampling technique. RESULTS: Seven variables, including demographic, lifestyle, and family history information, were examined. Multifactorial logistic regression analysis revealed that age [odds ratio (OR): 1.03, 95% confidence interval (CI): 1.02-1.03, P < 0.001], body mass index (BMI) (OR: 1.07, 95%CI: 1.06-1.08, P < 0.001), waist circumference (WC) (OR: 1.03, 95%CI: 1.02-1.03 P < 0.001), lifestyle (OR: 0.45, 95%CI: 0.42-0.48, P < 0.001), and family history (OR: 4.28, 95%CI: 4.04-4.54, P < 0.001) were the most significant predictors of high-risk CRC. Healthy lifestyle was a protective factor, whereas family history was the most significant risk factor. The area under the curve was 0.734 (95%CI: 0.723-0.745) for the final validation set ROC curve and 0.735 (95%CI: 0.728-0.742) for the training set ROC curve. The calibration curve demonstrated a high correlation between the CRC high-risk population predicted by the nomogram model and the actual CRC high-risk population. CONCLUSION: The early-screening nomogram model for CRC prediction in high-risk populations developed in this study based on age, BMI, WC, lifestyle, and family history exhibited high accuracy.

Adenosine A2A receptor antagonist KW6002 protects against A53T mutant alpha-synuclein-induced brain damage and neuronal apoptosis in Parkinson's disease mice by restoring autophagic flux.

BACKGROUND: Protein misfolding and inclusion body aggregation caused by α-Syn mutations in the brain often cause neurodegeneration and cognitive impairment, among which the A53T point mutation is more common. Inhibition of adenosine A2A receptor (A2AR) can alleviate the pathological symptoms of brain dysfunction caused by A53T-α-Syn protofibrils, but the mechanism of action is still unclear. AIM: This studies aimed to investigate the potential therapeutic role of the A2AR inhibitor KW6002 in a mouse model of brain synucleinopathy. METHODS: A53T-α-Syn fibre precursor cell nuclear protein was injected into the bilateral prefrontal cortex of mice to establish a synucleinopathy animal model, and the A2AR inhibitor KW6002 (5 mg/kg) was injected intraperitoneally to intervene. RESULT: The intracerebral injection of A53T-α-Syn protofibrils triggers the formation of inclusion bodies in the brain, leading to astrocyte activation, an increased number of apoptotic cells, and suppression of autophagic flux. The administration of KW6002 significantly reversed these phenomena. In vitro experiments revealed that A53T-α-Syn protofibrils inhibited HT-22 autophagy in mouse hippocampal neuronal cells, whereas KW6002 increased cellular autophagic flux, upregulated the expression of LAMP2A and Hsc70 proteins and inhibited the expression of SQSTM1 protein. The present study suggests that KW6002 reduces the level of α-Syn phosphorylation by inhibiting A2AR protein, at the same time, enhances the autophagic flux of neuronal cells, resulting in the degradation of A53T-α-Syn protofibrils and thus reducing the neuronal toxicity and apoptosis induced by A53T-α-Syn protofibrils. CONCLUSION: KW6002 has a significant protective effect on neuronal injury induced by A53T-α-Syn.

Associations between composite dietary antioxidant index and estimated 10-year atherosclerotic cardiovascular disease risk among U.S. adults.

Background: Atherosclerotic cardiovascular disease (ASCVD) remains the leading cause of death and disability both in U.S. and worldwide. Antioxidants have been proved critical in mitigating the development of atherosclerosis. This study aimed to investigate the associations between composite dietary antioxidant index (CDAI) and estimated 10-year ASCVD risk among U.S. adults. Methods: Data extracted from the National Health and Nutrition Examination Survey were analyzed. A total of 10,984 adults aged 18 years and above were included in this study. CDAI was calculated based on the dietary intake reported in their 24-h recall interviews. The estimated 10-year ASCVD risk was calculated via Pooled Cohort Equations (PCE). Results: After adjusting potential confounders, it was indicated that CDAI score was negatively correlated with 10-year ASCVD risk (OR 0.97, 95% CI 0.95-0.99). Stratify CDAI score by quartile, results showed that participants in the second, third, and fourth quartiles had lower ASCVD odds ratio (Q2: OR 0.87, 95% CI 0.69-1.09; Q3: OR 0.78, 95% CI 0.62-0.98; Q4: OR 0.74, 95% CI 0.59-0.94) than those in the first quartile (Q1, lowest CDAI score group), which was confirmed by the trend test as well (p < 0.05). Subgroup analyses stratified by sex, age, race/ethnicity, and smoking status did not show significant effect modification. Conclusion: Higher dietary antioxidants intake is associated with lower ASCVD risk among U.S. adults, for which policymakers and healthcare professionals may consider increasing the consumption of antioxidant-rich foods as a preventive strategy for ASCVD.

Kinase-independent role of mTOR and on-/off-target effects of an mTOR kinase inhibitor.

mTOR, as a serine/threonine kinase, is a widely pursued anticancer target. Multiple clinical trials of mTOR kinase inhibitors are ongoing, but their specificity and safety features remain lacking. Here, we have employed an inducible kinase-inactive D2338A mTOR knock-in mouse model (mTOR-/KI) together with a mTOR conditional knockout model (mTOR-/-) to assess the kinase-dependent/-independent function of mTOR in hematopoiesis and the on-/off-target effects of mTOR kinase inhibitor AZD2014. Despite exhibiting many similar phenotypes to mTOR-/- mice in hematopoiesis, the mTOR-/KI mice survived longer and showed differences in hematopoietic progenitor cells compared to mTOR-/- mice, suggesting a kinase-independent function of mTOR in hematopoiesis. Gene expression signatures in hematopoietic stem cells (HSCs) further revealed both kinase-dependent and independent effects of mTOR. AZD2014, a lead mTOR kinase inhibitor, appeared to work mostly on-target in suppressing mTOR kinase activity, mimicking that of mTOR-/KI HSCs in transcriptome analysis, but it also induced a small set of off-target responses in mTOR-/KI HSCs. In murine and human myeloid leukemia, besides kinase-inhibitory on-target effects, AZD2014 displayed similar off-target and growth-inhibitory cytostatic effects. These studies provide new insights into kinase-dependent/-independent effects of mTOR in hematopoiesis and present a genetic means for precisely assessing the specificity of mTOR kinase inhibitors.

Multimodal ultrasonography of male primary breast lymphoma.

① Gray scale ultrasound showed multiple hypoechoic masses of varying sizes in the right breast. The larger one was 1.8 × 0.7 cm (arrow), oval in shape, with clear boundaries, and lymphatic hilar-like structures. ② Color Doppler ultrasonography showed blood flow signals within the hypoechoic mass, and the larger (arrow) mass showed blood flow signals similar to lymphatic hilum. ③ Elastography showed the mass was soft and blue (short arrow) or green (long arrow) in texture, and the surrounding tissue was hard and red in texture. ④ Contrast-enhanced ultrasound showed that after 19 s of contrast agent injection, the whole breast showed a 'snowflake' high enhancement, but no enhancement was observed in local areas (arrow). ⑤ The ultrasound-guided puncture image clearly showed the puncture needle (arrow) insert into the hypoechoic mass for biopsy. ⑥ The arrow in the pathological image (HE, 20 × 10 times) showed tumor cells.

Loss of phosphatase CTDNEP1 potentiates aggressive medulloblastoma by triggering MYC amplification and genomic instability.

MYC-driven medulloblastomas are highly aggressive childhood brain tumors, however, the molecular and genetic events triggering MYC amplification and malignant transformation remain elusive. Here we report that mutations in CTDNEP1, a CTD nuclear-envelope-phosphatase, are the most significantly enriched recurrent alterations in MYC-driven medulloblastomas, and define high-risk subsets with poorer prognosis. Ctdnep1 ablation promotes the transformation of murine cerebellar progenitors into Myc-amplified medulloblastomas, resembling their human counterparts. CTDNEP1 deficiency stabilizes and activates MYC activity by elevating MYC serine-62 phosphorylation, and triggers chromosomal instability to induce p53 loss and Myc amplifications. Further, phosphoproteomics reveals that CTDNEP1 post-translationally modulates the activities of key regulators for chromosome segregation and mitotic checkpoint regulators including topoisomerase TOP2A and checkpoint kinase CHEK1. Co-targeting MYC and CHEK1 activities synergistically inhibits CTDNEP1-deficient MYC-amplified tumor growth and prolongs animal survival. Together, our studies demonstrate that CTDNEP1 is a tumor suppressor in highly aggressive MYC-driven medulloblastomas by controlling MYC activity and mitotic fidelity, pointing to a CTDNEP1-dependent targetable therapeutic vulnerability.

CircRTN1 stimulates HMGB1 to regulate the malignant progression of papillary thyroid cancer by sponging miR-101-3p.

BACKGROUND: The important role played by circular RNA (circRNA) in promoting the progression of papillary thyroid cancer (PTC) is attracting ever more attention among medical researchers. However, what the precise contribution is of circRTN1 in PTC progression remains unclear. The study was designed to analyze the role and mechanism of circRTN1 in regulating PTC progression. METHODS: Human PTC cell lines (TPC-1 and IHH-4) and human thyroid normal cells (Nthy-ori 3-1) were used for in vitro assays. mRNA or protein expression of circRTN1, miR-101-3p, and high mobility group box 1 (HMGB1) were detected by quantitative real-time polymerase chain reaction or western blot. Cell proliferation was investigated by cell counting kit-8 assay, cell colony formation assay, and 5-ethynyl-2'-deoxyuridine assay. Wound-healing assay and transwell invasion assay were conducted to evaluate cell migration and invasion. Dual-luciferase reporter assay and RNA immunoprecipitation assay were applied to verify the target relations between circRTN1, miR-101-3p, and HMGB1. A xenograft tumor model was established to demonstrate the effect of circRTN1 on tumor formation in vivo. An immunohistochemistry assay was used to detect protein expression of HMGB1, ki-67, E-cadherin, and vimentin. RESULTS: In comparison with healthy thyroid tissues and cells, PTC tissues and cells displayed high circRTN1 RNA expression and high HMGB1 mRNA and protein expression but low miR-101-3p expression. Silencing of circRTN1 suppressed PTC cell proliferation, migration, and invasion in vitro. MiR-101-3p was a target of circRTN1, and the knockdown of miR-101-3p relieved circRTN1 absence-mediated suppressive effects on PTC cell malignancy. HMGB1 was identified as a target gene of miR-101-3p, and overexpressed HMGB1 almost reverted the inhibitory impacts induced by miR-101-3p mimic in PTC cells. Moreover, circRTN1 silencing hampered tumor formation in vivo. CONCLUSION: CircRTN1 depletion impeded PTC cell malignancy via the miR-101-3p/HMGB1 pathway, which provided a possible circRNA-targeted therapeutic strategy for PTC.

miR-204 suppresses uveal melanoma cell migration and invasion through negative regulation of RAB22A.

Uveal melanoma (UM), a frequently seen adulthood primary ocular malignancy, shows high aggressiveness. Accumulating studies have revealed the crucial effects of microRNAs (miRNAs) on tumorigenesis and development in various human tumors. miR-204, the cancer-associated miRNA, shows dysregulation and is related to several human malignancies, but its effect on UM remains unknown. The present work focused on exploring miR-204's effect on UM and elucidating its possible molecular mechanisms. According to our results, miR-204 expression markedly increased within both UM tissues and cell lines. As revealed by functional analysis, miR-204 suppressed UM cell invasion and migration. Besides, RAB22A expression decreased through directly binding miR-204 into the corresponding 3' untranslated region (3'UTR) in UM cells. Furthermore, the RAB22A mRNA level increased, which was negatively related to the miR-204 level within UM samples. As revealed by mechanical research, miR-204 exerted its inhibition on the invasion and migration of UM cells via RAB22A. Taken together, this study suggested the tumor-suppressing effect of miR-204 on UM through down-regulating RAB22A. Thus, miR-204 may serve as the new anti-UM therapeutic target.

The relationship among pregnancy-related anxiety, perceived social support, family function and resilience in Chinese pregnant women: a structural equation modeling analysis.

BACKGROUND: Accumulating evidence suggests that pregnancy-related anxiety (PRA) has adverse impacts on maternity health and infant development. A substantial body of literature has documented the important influence of family function, perceived social support and resilience on PRA. However, research identifying the mediating mechanisms underlying this relationship in China are still lacking. Therefore, the current study aimed to investigate the prevalence of PRA under the three-child policy in China, and also explore the interrelationships among perceived social support, family function, resilience, and PRA. METHODS: In this cross-sectional study, a convenient sampling method was used to select 579 pregnant women who underwent prenatal examination at the maternity outpatient departments of the First Affiliated Hospital of Chongqing Medical University in China from December 2021 to April 2022. Participants were required to complete the following questionnaires: the demographic form, the Chinese Pregnancy-related Anxiety scale, the 10-item Connor-Davidson Resilience Scale, the APGAR Family Care Index Scale, and Multidimensional Scale of Perceived Social Support. Pearson correlation analysis was utilized to examine the rudimentary relationship among the study variables. Bootstrapping analyses in the structural equation modeling were applied to identify the significance of indirect effects. RESULTS: There were 41.4% of pregnant Chinese women indicating PRA. Correlational analyses indicated that perceived social support, family function and resilience were negatively associated with PRA (r = - 0.47, P < 0.01; r = - 0.43, P < 0.01; r = - 0.37, P < 0.01, respectively). The results of bootstrapping analyses demonstrated significant indirect effects of perceived social support (ß = - 0.098, 95% CI [- 0.184, - 0.021]) and family function (ß = - 0.049, 95% CI [- 0.103, - 0.011]) on PRA via resilience. CONCLUSIONS: Chinese pregnant women are suffering from high levels of PRA. Better family function and perceived social support might reduce the occurrence of PRA, as well as by the mediating effects of resilience. Healthcare providers must be concerned about PRA and perform corresponding actions to reduce it. By strengthening social support and improving family function, antenatal care providers could effectively reduce or prevent PRA. And more importantly, implementing resilience-promoting measures are also essential to relieve anxiety and support mental health in pregnant women.

PRMT3 drives glioblastoma progression by enhancing HIF1A and glycolytic metabolism.

Glioblastoma (GBM) is the most common and aggressive primary brain tumor, but the mechanisms underlying tumor growth and progression remain unclear. The protein arginine methyltransferases (PRMTs) regulate a variety of biological processes, however, their roles in GBM growth and progression are not fully understood. In this study, our functional analysis of gene expression networks revealed that among the PRMT family expression of PRMT3 was most significantly enriched in both GBM and low-grade gliomas. Higher PRMT3 expression predicted poorer overall survival rate in patients with gliomas. Knockdown of PRMT3 markedly reduced the proliferation and migration of GBM cell lines and patient-derived glioblastoma stem cells (GSC) in cell culture, while its over-expression increased the proliferative capacity of GSC cells by promoting cell cycle progression. Consistently, stable PRMT3 knockdown strongly inhibited tumor growth in xenograft mouse models, along with a significant decrease in cell proliferation as well as an increase in apoptosis. We further found that PRMT3 reprogrammed metabolic pathways to promote GSC growth via increasing glycolysis and its critical transcriptional regulator HIF1α. In addition, pharmacological inhibition of PRMT3 with a PRMT3-specific inhibitor SGC707 impaired the growth of GBM cells. Thus, our study demonstrates that PRMT3 promotes GBM progression by enhancing HIF1A-mediated glycolysis and metabolic rewiring, presenting a point of metabolic vulnerability for therapeutic targeting in malignant gliomas.

Precise dapagliflozin delivery by cardiac homing peptide functionalized mesoporous silica nanocarries for heart failure repair after myocardial infarction.

Myocardial infarction (MI) may cause irreversible damage or destroy to part of the heart muscle, affecting the heart's ability and power to pump blood as efficiently as before, often resulting in heart failure (HF). Cardiomyocyte death and scar formation after MI may then trigger chronic neurohormonal activation and ventricular remodeling. We developed a biocompatible and mono-dispersed mesoporous silica nanoparticles (MSN) divergent porous channel for dapagliflozin (DAPA) loading. After surface modification of the cardiac-targeting peptides, the novel drug delivery system was successfully homed, and precisely released drugs for the hypoxic and weak acid damaged cardiomyocytes. Our biocompatible MSN- based nanocarriers for dapagliflozin delivery system could effective cardiac repair and regeneration in vivo, opening new opportunities for healing patients with ischemic heart disease in clinical.

Prognostic value of nutritional status in patients with human immunodeficiency virus infection-related lymphoma.

Objective: To investigate the predictive value of nutritional status on the prognosis of patients with human immunodeficiency virus (HIV) infection-related lymphoma. Materials and methods: A total of 149 patients with HIV infection-related lymphoma who were admitted to our hospital from August 2012 to May 2022 were selected as research subjects. Based on the patient prognosis, they were divided into a poor prognosis group (n = 30) and a good prognosis group (n = 119). General data from patients in both groups were collected, and the nutritional status of the patients was evaluated using the Controlling Nutritional Status (CONUT) score. Factors affecting the prognosis of HIV infection-related lymphoma were analyzed using univariate and multivariate analyses, and a prediction model was developed based on the analyzed factors. The receiver operating characteristic (ROC) curve was used to analyze the prediction model of the CONUT score alone and included the CONUT score in the prognosis of patients with HIV infection-related lymphoma. The predictive value of the data was assessed, and a survival curve was drawn to compare the survival of patients with different nutritional statuses. Results: There were significant differences in age, B symptoms, treatment conditions, International Prognostic Index (IPI), pathological stage, Eastern Collaborative Tumor Group physical status score (ECOG PS), CD4+ cell count, ß2 microglobulin, and lactate dehydrogenase (LDH) between the poor prognosis group and the good prognosis group (p < 0.05). The CONUT score of the poor prognosis group was higher than that of the good prognosis group, and the difference was statistically significant (p < 0.05). A univariate analysis demonstrated that the age, B symptoms, treatment status, IPI, pathological stage, ECOG PS, CD4+ cell count, ß2 microglobulin, LDH, and CONUT score were prognostic factors for patients with HIV infection-related lymphoma (p < 0.05). The results of a multivariate regression analysis demonstrated that the age, B symptoms, treatment status, IPI, pathological stage, ECOG PS, and CONUT score were independent risk factors for the prognosis of patients with HIV infection-related lymphoma (p < 0.05). The prediction model was constructed according to the multivariate Cox regression analysis results. The model formula was as follows: Logit(p) = -10.687 + 1.728 × age + 1.713 × B symptoms + 1.682 × treatment status + 1.810 × IPI + 1.643 × pathological stage + 1.584 × ECOG PS + 1.779 × CONUT score. The ROC curve was used to analyze the predictive value of the CONUT score alone and the predictive model including the CONUT score on the prognosis of patients with HIV infection-related lymphoma. The predictive value of the prognosis of patients with tumors was higher (p < 0.05). According to the results of the ROC curve analysis, the patients were divided into a high CONUT group (CONUT > 6.00 points, n = 31) and a low CONUT group (CONUT ≤ 6.00 points, n = 118) based on the Optimum threshold of the CONUT score. The survival curve showed that the survival rate of the high CONUT group was lower than that of the low CONUT group (p < 0.05). Conclusion: The poor prognosis of HIV infection-related lymphoma is related to nutritional status, which is an independent risk factor affecting the prognosis of patients and can be used as a practical indicator to predict the prognosis of patients.

Human fetal cerebellar cell atlas informs medulloblastoma origin and oncogenesis.

Medulloblastoma (MB) is the most common malignant childhood brain tumour1,2, yet the origin of the most aggressive subgroup-3 form remains elusive, impeding development of effective targeted treatments. Previous analyses of mouse cerebella3-5 have not fully defined the compositional heterogeneity of MBs. Here we undertook single-cell profiling of freshly isolated human fetal cerebella to establish a reference map delineating hierarchical cellular states in MBs. We identified a unique transitional cerebellar progenitor connecting neural stem cells to neuronal lineages in developing fetal cerebella. Intersectional analysis revealed that the transitional progenitors were enriched in aggressive MB subgroups, including group 3 and metastatic tumours. Single-cell multi-omics revealed underlying regulatory networks in the transitional progenitor populations, including transcriptional determinants HNRNPH1 and SOX11, which are correlated with clinical prognosis in group 3 MBs. Genomic and Hi-C profiling identified de novo long-range chromatin loops juxtaposing HNRNPH1/SOX11-targeted super-enhancers to cis-regulatory elements of MYC, an oncogenic driver for group 3 MBs. Targeting the transitional progenitor regulators inhibited MYC expression and MYC-driven group 3 MB growth. Our integrated single-cell atlases of human fetal cerebella and MBs show potential cell populations predisposed to transformation and regulatory circuitries underlying tumour cell states and oncogenesis, highlighting hitherto unrecognized transitional progenitor intermediates predictive of disease prognosis and potential therapeutic vulnerabilities.

Nursing Skill Assessment of Hospital Nurses in Management of Critically Ill Patients.

Objective: To analyze the application of standardized nursing procedures in critically ill patients' nursing evaluation. Methods: 90 cases of critically ill patients aged from 18 to 65 who were treated in our hospital from April 2020 to April 2021 were selected and divided into the control group and observation group, respectively, with 45 cases according to the drawing method. The rescue time, blood pressure, heart rate before and after nursing, adverse mood, length of stay, incidence of adverse events, ICU transfer and death, and satisfaction of 2 groups were statistically analyzed and compared. Results: The rescue time of cardiopulmonary resuscitation, oxygen inhalation, venous opening, and endotracheal resuscitation in the observation group was 3.24 ± 1.01, which is lower than that in the control group, 6.65 ± 2.11, with statistical significance (P < 0.05). Similarly, the vital signs in the observation group were 2.45 ± 0.44, which is also significantly lower than that in the control group, 5.67 ± 1.56. After nursing, the blood pressure and heart rate in the observation group were lower than those in control group, with statistical significance (P < 0.05). The adverse mood of the observation group after nursing was lower than that of the control group, with statistical significance (P < 0.05). The length of stay, incidence of adverse events, intensive care unit (ICU) transfer, and death in the observation group were lower than those in the control group, with statistical significance (P < 0.05). The length of stay in the observation group was 8.87 ± 2.11, while 11.34 ± 2.45 in the observation group. The incidence of adverse events in the observation group was 1, while 8 in the observation group. The length of stay in the observation group was 8.87 ± 2.11, while 11.34 ± 2.45 in the observation group. The ICU transfer in the observation group were 2, while 9 in the observation group. There was no death in the observation group, however, 4 in the observation group. Nursing satisfaction in the observation group was higher than that in the control group, with statistical significance (P < 0.05). The number of patients that are very satisfied in the observation group was 28, while 20 in the control group. The number of patients that are satisfied in the observation group was the same as in the control group, both 15. However, the number of patients that are dissatisfied in the observation group was 2, while 10 in the control group. Conclusion: The application of the standardized nursing process in the nursing of critically ill patients can not only effectively reduce the self-rating anxiety scale (SAS) and sarcoidosis diagnostic score (SDS) of patients but also reduce the incidence of complications and improve the nursing satisfaction of patients.

Conserved and Distinct Functions of the Autism-Related Chromatin Remodeler CHD8 in Embryonic and Adult Forebrain Neurogenesis.

The chromatin remodeler CHD8 represents a high-confidence risk factor in autism, a multistage progressive neurologic disorder, however the underlying stage-specific functions remain elusive. In this study, by analyzing Chd8 conditional knock-out mice (male and female), we find that CHD8 controls cortical neural stem/progenitor cell (NSC) proliferation and survival in a stage-dependent manner. Strikingly, inducible genetic deletion reveals that CHD8 is required for the production and fitness of transit-amplifying intermediate progenitors (IPCs) essential for upper-layer neuron expansion in the embryonic cortex. p53 loss of function partially rescues apoptosis and neurogenesis defects in the Chd8-deficient brain. Further, transcriptomic and epigenomic profiling indicates that CHD8 regulates the chromatin accessibility landscape to activate neurogenesis-promoting factors including TBR2, a key regulator of IPC neurogenesis, while repressing DNA damage- and p53-induced apoptotic programs. In the adult brain, CHD8 depletion impairs forebrain neurogenesis by impeding IPC differentiation from NSCs in both subventricular and subgranular zones; however, unlike in embryos, it does not affect NSC proliferation and survival. Treatment with an antidepressant approved by the Federal Drug Administration (FDA), fluoxetine, partially restores adult hippocampal neurogenesis in Chd8-ablated mice. Together, our multistage functional studies identify temporally specific roles for CHD8 in developmental and adult neurogenesis, pointing to a potential strategy to enhance neurogenesis in the CHD8-deficient brain.SIGNIFICANCE STATEMENT The role of the high-confidence autism gene CHD8 in neurogenesis remains incompletely understood. Here, we identify a stage-specific function of CHD8 in development of NSCs in developing and adult brains by conserved, yet spatiotemporally distinct, mechanisms. In embryonic cortex, CHD8 is critical for the proliferation, survival, and differentiation of both NSC and IPCs during cortical neurogenesis. In adult brain, CHD8 is required for IPC generation but not the proliferation and survival of adult NSCs. Treatment with FDA-approved antidepressant fluoxetine partially rescues the adult neurogenesis defects in CHD8 mutants. Thus, our findings help resolve CHD8 functions throughout life during embryonic and adult neurogenesis and point to a potential avenue to promote neurogenesis in CHD8 deficiency.

ANGPTL4 regulates the osteogenic differentiation of periodontal ligament stem cells.

The molecular mechanism of mechanical force regulating the osteogenic differentiation of periodontal ligament stem cells (PDLSCs) has not been clearly elucidated. In this study, two mRNA-seqs, GSE106887 and GSE109167, which contained several samples of PDLSCs under mechanical force, were downloaded from Gene Expression Omnibus. Differential expression analysis was firstly taken between GSE106887 and GSE109167, then the common 84 up-regulated genes and 26 down-regulated genes were selected. Function enrichment analysis was used to identify the key genes and pathways in PDLSCs subjected to the tension and compression force. PDLSCs were isolated from human periodontal ligament tissues. The effects of ANGPTL4 knockdown with shRNA on the osteogenic differentiation of PDLSCs were studied in vitro. Then, the orthodontic tooth movement (OTM) rat model was used to study the expression of HIF-1α and ANGPTL4 in alveolar bone remodeling in vivo. ANGPTL4 and the HIF-1 pathway were identified in PDLSCs subjected to the tension and compression force. alizarin red staining, alcian blue staining, and oil red O staining verified that PDLSCs had the ability of osteogenic, chondrogenic, and adipogenic differentiation, respectively. Verification experiment revealed that the expression of ANGPTL4 in PDLSCs significantly increased when cultured under osteogenic medium in vitro. While ANGPTL4 was knocked down by shRNA, the levels of ALPL, RUNX2, and OCN decreased significantly, as well as the protein levels of COL1A1, ALPL, RUNX2, and OCN. During the OTM, the expression of HIF-1α and ANGPTL4 in periodontal ligament cells increased on the tension and compression sides. We concluded the positive relationship between ANGPTL4 and osteogenic differentiation of PDLSCs.

Outcome of teeth restored with CAD/CAM zirconium dioxide post-cores: a retrospective study with a follow-up period of 3-6 years.

STATEMENT OF PROBLEM: Computer aided design/computer aided manufacturing (CAD/CAM) zirconia post-cores is one of the options of post crown restoration materials due to their esthetic properties and superior mechanical strength. However, the clinical effect on aesthetics and strength properties is unclear due to the lack of results of their long-term follow-up. PURPOSE: This retrospective clinical study aims to analyze the survival rate, clinical manifestations, and failure factors after CAD/CAM zirconia post-core restoration. MATERIAL AND METHODS: Clinical and radiographic examinations were performed on 342 patients with 400 teeth for 3-6 years postsurgical follow-up examination. The patients were all received CAD/CAM zirconia post-cores and all-ceramic crowns at the Department of Prosthodontics in the public hospital. The retrospective outcomes were conducted after zirconia post restoration, including survival rate by Kaplan-Meier analysis and findings of manifestations and failure factors. The effects of gender and dental position on survival rate were analyzed by Cox-Mantel Test. RESULTS: This study retrospectively evaluated 261 teeth from 229 patients with a 35% drop-out rate. The survival rate was 96.0%, and the success rate was 92.4%. According to the tooth position classification, the survival rate was 100% for 101 anterior teeth, 95.4% for 69 premolars, and 88.3% for 91 molars. According to gender, the survival rate of the male group was 92.3%, while that of the female group was 98.0%, with a significant difference (P < 0.01). The complications included crown fracture (1.9%) periapical inflammation (1.9%), crown debonding (1.1%), percussion abnormal (1.9%) and root fracture (0.8%). CONCLUSIONS: Within the limitations of this retrospective study, it can be concluded that CAD/CAM zirconia post-cores are clinically promising. Compared with the posterior teeth, CAD/CAM zirconia post-cores are more suitable for anterior teeth.

A platform method for plasmid isoforms analysis by capillary gel electrophoresis.

In the production of novel biological products, plasmids are often engineered into delivery vectors for target genes, which can be used directly as vaccines or as intermediate products for gene/cell therapy. Plasmid DNA exists in several topological forms such as supercoiled, linear, and open circular. As supercoiled plasmid shows the highest efficiency in transfecting eukaryotic cells, the content of supercoiled plasmids becomes an important indicator of plasmid quality. CGE is an effective analysis method for separating different topological structures of plasmids. For the purpose of providing plasmid manufacturers and regulatory agencies with an efficient and readily used tool for monitoring the quality of plasmids, this article identifies the optimal separation and detection conditions of CGE, presents a platform-based plasmid analytical method, and uses plasmid of different sizes to verify the feasibility of this method. In terms of detector, the LIF detector has obvious advantages over the ultraviolet detector in sensitivity and resolution. Using the optimal CE condition (10× gel buffer), baseline separation of different topological forms and impurities can be achieved for different plasmid sizes (5.9, 7.8, 15.4 kb). In addition, 6.5 kb plasmid was used to compare the different separation technologies such as CGE-LIF, ion exchange chromatography and agarose gel electrophoresis. The result shows that CGE-LIF can provide better resolution and quantitation accuracy than ion exchange chromatography and agarose gel electrophoresis. CGE-LIF, as a quick and convenient method to separate and quantify plasmids, has the advantages of high sensitivity, high resolution, and high quantitative accuracy. Therefore, it is ideal for analysis of plasmids with different sizes, and it can also be used as a platform method for manufacturers and regulatory agencies to monitor the purity and stability of plasmids.

Tumour-suppressing functions of the lncRNA MBNL1-AS1/miR-889-3p/KLF9 axis in human breast cancer cells.

This study aimed to explore the role and potential mechanism of the long non-coding (lncRNA) MBNL1-AS1 in human breast cancer. We included 80 patients with breast cancer in this study. Breast cancer cell lines, including MCF7, SKBR3, MDA-MB-231 and MDA-MB-415, and the normal human breast cell line MCF10A were used in this study. MBNL1-AS1, miR-889-3p mimics, si-Krüppel-like factor 9 (KLF9) or their controls were transfected in the cells. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry assay were performed to detect the expression of MBNL1-AS1, miR-889-3p and KLF9. Cell proliferation, invasion and migration were detected. Luciferase reporter gene and pull-down assay were performed to verify the target relationship among MBNL1-AS1, miR-889-3p and KLF9. Glycolysis was also detected after transfection. The expression of the lncRNA MBNL1-AS1 was low in the breast cancer tissues and cells. Lower expression levels of the lncRNA MBNL1-AS1 were associated with poor prognosis of breast cancer. Overexpression of the lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer cells. The lncRNA MBNL1-AS1 could interact with miR-889-3p, and KLF9 was the downstream target of miR-889-3p. Moreover, miR-889-3p was negatively correlated with KLF9 and lncRNA MBNL1-AS1. Both miR-889-3p and si-KLF9 could reverse the overexpression of lncRNA MBNL1-AS1 in breast cancer development. The lncRNA MBNL1-AS1 decreased proliferation, invasion, migration and glycolysis of breast cancer via the miR-889-3p/KLF9 axis, which might be a potential biomarker for the diagnosis of breast cancer.