Iron-Confined CRISPR/Cas9-Ribonucleoprotein Delivery System for Redox-Responsive Gene Editing.

Clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 9 (Cas9) is a promising gene editing tool to treat diseases at the genetic level. Nonetheless, the challenge of the safe and efficient delivery of CRISPR/Cas9 to host cells constrains its clinical applicability. In the current study, a facile, redox-responsive CRISPR/Cas9-Ribonucleoprotein (RNP) delivery system by combining iron-coordinated aggregation with liposomes (Fe-RNP@L) is reported. The Fe-RNP is formed by the coordination of Fe3+ with amino and carboxyl groups of Cas9, which modifies the lipophilicity and surface charge of RNP and alters cellular uptake from primary endocytosis to endocytosis and cholesterol-dependent membrane fusion. RNP can be rapidly and reversibly released from Fe-RNP in response to glutathione without loss of structural integrity and enzymatic activity. In addition, iron coordination also improves the stability of RNP and substantially mitigates cytotoxicity. This construct enabled highly efficient cytoplasmic/nuclear delivery (≈90%) and gene-editing efficiency (≈70%) even at low concentrations. The high payload content, high editing efficiency, good stability, low immunogenicity, and ease of production and storage, highlight its potential for diverse genome editing and clinical applications.

Inhibition of FoxO1 alleviates polycystic ovarian syndrome by reducing inflammation and the immune response.

The aim of this study was to explore the role of forkhead box transcription Factor O1 (FoxO1) in chronic inflammation in polycystic ovary syndrome (PCOS). A PCOS rat model was constructed as an in vivo model by letrozole induction, and granulosa cells (GCs) from PCOS rats were isolated and cultured as an in vitro cellular model. FoxO1 was knocked down by shRNA and siRNA in the PCOS rat model and GCs model, respectively. H&E staining was conducted to evaluate the effect of FoxO1 inhibition on ovarian pathology and dysfunction in PCOS rats. The levels of inflammatory cytokines in the ovaries and uterus of PCOS rats and in GCs were assessed by ELISA. Flow cytometry was used to evaluate the changes in the contents of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats. CCK-8 assays and Annexin V-FITC/PI staining were performed to evaluate the proliferation and apoptosis of GCs. The expression of genes and proteins related to the TLR4/NF-κB/NLRP3 pathway in GCs was determined by RT-qPCR and Western blotting. The results indicated that FoxO1 was highly expressed in PCOS rat model. Inhibition of FoxO1 significantly mitigated the pathological changes and dysfunction in the ovaries of PCOS rats while also suppressing inflammation and fibrosis in the ovaries and uterus. Moreover, knocking down FoxO1 facilitated the restoration of the normal ratio of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats and promoted M2 polarization of macrophages. Additionally, inhibition of FoxO1 promoted the proliferation of GCs and inhibited the inflammatory response in GCs. Furthermore, FoxO1 knockdown inhibited the activation of the NF-κB pathway and the formation of the NLRP3 inflammasome in GCs. In conclusion, inhibition of FoxO1 can alleviate PCOS by inhibiting the TLR4/NF-κB/NLRP3 pathway to reduce inflammation and the immune response.

Clinical characteristics, risk factors, and outcomes of patients with polymicrobial Pseudomonas aeruginosa bloodstream infections.

BACKGROUND: Previous studies on polymicrobial Pseudomonas aeruginosa bloodstream infections (Pa-BSIs) are dated, and it is necessary to reanalyze polymicrobial Pa-BSIs. The aim of this study was to investigate clinical characteristics and risk factors for polymicrobial Pa-BSI in comparison with monomicrobial Pa-BSI. METHODS: A double-center retrospective observational study was performed between January 1, 2013 and June 30, 2022, in two tertiary hospitals. All patients with Pa-BSI were enrolled, and their clinical data were collected by reviewing electronic medical records. RESULTS: A total of 278 patients with Pa-BSI were enrolled, including 77 patients (27.7%) with polymicrobial Pa-BSI. Compared with monomicrobial Pa-BSI, the main source of polymicrobial Pa-BSI was pneumonia (49.4% vs 31.3%, p < 0.01), whereas the main source of monomicrobial Pa-BSI was primary BSIs (21.9% vs 2.6%, p = 0.04). In multivariate analysis, a history of cerebrovascular accident (CVA) (adjusted odds ratio [OR], 3.62; 95% CI, 1.46-8.92) was independently associated with polymicrobial Pa-BSI. Primary BSI was associated with monomicrobial Pa-BSI (OR, 0.08; 95% CI, 0.02-0.38). Patients with polymicrobial Pa-BSI had a longer intensive care unit (ICU) length of stay after onset of BSI than those with monomicrobial Pa-BSI (2 [2, 16] vs 13 [3.75, 29], p = 0.02). CONCLUSION: Patients with Pa-BSI and the presence of CVA need to be alert to the possibility of polymicrobial BSI occurrence. Prolonged ICU stay and pneumonia as a source of BSI warrant clinician attention for polymicrobial Pa-BSI, and primary BSIs are likely associated with monomicrobial BSIs.

Histidine modulates amyloid-like assembly of peptide nanomaterials and confers enzyme-like activity.

Amyloid-like assembly is not only associated with pathological events, but also leads to the development of novel nanomaterials with unique properties. Herein, using Fmoc diphenylalanine peptide (Fmoc-F-F) as a minimalistic model, we found that histidine can modulate the assembly behavior of Fmoc-F-F and induce enzyme-like catalysis. Specifically, the presence of histidine rearranges the ß structure of Fmoc-F-F to assemble nanofilaments, resulting in the formation of active site to mimic peroxidase-like activity that catalyzes ROS generation. A similar catalytic property is also observed in Aß assembled filaments, which is correlated with the spatial proximity between intermolecular histidine and F-F. Notably, the assembled Aß filaments are able to induce cellular ROS elevation and damage neuron cells, providing an insight into the pathological relationship between Aß aggregation and Alzheimer's disease. These findings highlight the potential of histidine as a modulator in amyloid-like assembly of peptide nanomaterials exerting enzyme-like catalysis.

Combine with RNA-seq Reveals the Effect of Melatonin in the Synthesis of Melanin in Primary Melanocytes of Silky Fowls Black-Bone Chicken.

(1) Background: It was found that the melanin of black-bone chicken has various effects such as scavenging DPPH free radicals and anti-oxidation, and the synthesis of melanin is affected by various factors including hormones. In addition, several studies have found that melatonin affects the melanoma cell synthesis of melanin, which has not been reported in chicken primary melanocytes; so, relevant studies were conducted. (2) Methods: In this study, chicken primary melanocytes were isolated and characterized, and then melanocytes were treated with different concentrations of melatonin to investigate the effects of melatonin on melanin synthesis in chicken melanocytes in terms of melanin synthesis-related genes, melanin content, and tyrosinase activity, and combined with RNA seq to detect the change in gene expression level of chicken melanocytes after melatonin treatment. (3) Results: We isolated and characterized primary melanocytes, and indirect immunofluorescence assay results showed positive melanocyte marker genes. RT-qPCR results showed that melatonin decreased the expression of melanin synthesis-related genes. In addition, melatonin reduced the melanin content and decreased the tyrosinase activity of melanocytes in the treated group. A total of 1703 differentially expressed genes were screened by RNA-seq, and in addition, in the KEGG results, the signaling pathway associated with melanin synthesis, and the mTOR signaling pathway were enriched. (4) Conclusions: Melatonin could decrease the synthesis of melanin in chicken primary melanocytes.

The First Crested Duck Genome Reveals Clues to Genetic Compensation and Crest Cushion Formation.

The Chinese crested (CC) duck is a unique indigenous waterfowl breed, which has a crest cushion that affects its survival rate. Therefore, the CC duck is an ideal model to investigate the genetic compensation response to maintain genetic stability. In the present study, we first generated a chromosome-level genome of CC ducks. Comparative genomics revealed that genes related to tissue repair, immune function, and tumors were under strong positive selection, indicating that these adaptive changes might enhance cancer resistance and immune response to maintain the genetic stability of CC ducks. We also assembled a Chinese spot-billed (Csp-b) duck genome, and detected the structural variations (SVs) in the genome assemblies of three ducks (i.e., CC duck, Csp-b duck, and Peking duck). Functional analysis revealed that several SVs were related to the immune system of CC ducks, further strongly suggesting that genetic compensation in the anti-tumor and immune systems supports the survival of CC ducks. Moreover, we confirmed that the CC duck originated from the mallard ducks. Finally, we revealed the physiological and genetic basis of crest traits and identified a causative mutation in TAS2R40 that leads to crest formation. Overall, the findings of this study provide new insights into the role of genetic compensation in adaptive evolution.

Circ_PIAS1 Promotes the Apoptosis of ALV-J Infected DF1 Cells by Up-Regulating miR-183.

(1) Background: circRNAs are closed circular molecules with covalent bonds generated by reverse shearing, which have high stability and have different manifestations in different tissues, cells, or physiological conditions and play important roles in various disease processes and physiological processes. In addition, circ_PIAS1 has been screened out and verified, and the bioinformatics analyzed in previous studies. In this study, we investigated the function of circ_PIAS1 and studied its role in ALV-J infection to provide a basis for the role of circRNA in ALV-J infection. (2) Methods: the effect of circ_PIAS1 on apoptosis during ALV-J infection was studied by flow cytometry and detection of apoptotic gene expression, and miR-183 was screened by a biotin-labeled RNA pull-down technique. After overexpression and inhibition of miR-183, the effect of miR-183 on apoptosis in the process of ALV-J infection was studied by flow cytometry and detection of apoptotic gene expression. (3) Results: after overexpression of circ_PIAS1, flow cytometry and apoptotic gene expression showed that circ_PIAS1 promoted apoptosis. The results of RNA pull-down showed that 173 miRNAs could bind to circ_PIAS1, and circ_PIAS1 up-regulated the expression of miR-183. On the other hand, the same results were obtained whether miR-183 was overexpressed or inhibited that miR-183 affected ALV-J infection by promoting cell apoptosis. (4) Conclusions: circ_PIAS1 up-regulated the expression of miR-183 and influenced ALV-J infection by promoting cell apoptosis.

Evaluation of non-invasive gene detection in preimplantation embryos: a systematic review and meta-analysis.

BACKGROUND: Genetic abnormalities in embryos are responsible for most miscarriages and repeated embryo implantation failures, so a reliable preimplantation genetic screening method is urgently needed. Non-invasive preimplantation genetic testing (niPGT) is a potential method for embryo genetic diagnosis. However, the value of its application is controversial. This meta-analysis aimed to investigate and validate the diagnostic value of niPGT in patients undergoing in vitro fertilization (IVF). METHODS: This review used the "Preferred Reporting Items" as a systematic review and meta-analysis of the diagnostic test accuracy (PRISMA-DTA) statement. We searched PubMed, Embase, Web of Science Core Collection, and Cochrane Library up to May 2022 to retrieve non-invasive preimplantation gene detection studies. The eligible research quality was evaluated following the quality assessment study-2 system for diagnostic accuracy. The pooled receiver operator characteristic curve (SROC) and the area under SROC (AUC) were used to evaluate diagnostic performance quantitatively. Threshold effect, subgroup analysis, and meta-regression analysis were used to explore the source of heterogeneity. Deeks' funnel plots and sensitivity analyses were used to test the publication bias and stability of the meta-analysis, respectively. FINDINGS: Twenty studies met the inclusion criteria. The pooled sensitivity, specificity, and AUC were 0.84 (95% CI 0.72-0.91), 0.85 (95% CI 0.74-0.92), and 0.91 (95% CI 0.88-0.93), respectively. Subgroup analysis showed that the spent culture medium (SCM) subgroup had higher sensitivity and lower specificity than the SCM combined with the blastocoel fluid (BF) subgroup. Subgroup analysis showed that the study sensitivity and specificity of < 100 cases were higher than those of ≥ 100. Heterogeneity (chi-square) analysis revealed that sample size might be a potential source of heterogeneity. Sensitivity analysis and Deeks' funnel plots indicated that our results were relatively robust and free from publication bias. INTERPRETATION: The present meta-analysis indicated that the pooled sensitivity, specificity, and AUC of niPGT in preimplantation genetic testing were 0.84, 0.85, and 0.91, respectively. niPGT may have high detection accuracy and may serve as an alternative model for embryonic analysis. Additionally, by subgroup analysis, we found that BF did not improve the accuracy of niPGT in embryos. In the future, large-scale studies are needed to determine the detection value of niPGT.

Impact of Visceral Obesity on Structural and Functional Alterations of Gut Microbiota in Polycystic Ovary Syndrome (PCOS): A Pilot Study Using Metagenomic Analysis.

Objective: We aimed to identify structural and functional alterations of gut microbiota associated with visceral obesity in adult women with polycystic ovary syndrome (PCOS). Methods: Twenty-seven adults with PCOS underwent stool and fasting blood collection, oral glucose tolerance testing, and visceral fat area (VFA) measurement via dual-bioimpedance technique. Metagenomic analysis was used to analyze gut microbiota. Results: PCOS patients were divided into three groups: visceral obesity group (PCOS-VO, n=9, age 28.33±5.68 years, BMI 37.06±4.27 kg/m2, VFA 128.67±22.45 cm2), non-visceral obesity group (PCOS-NVO, n=10, age 25.40±4.53, BMI 30.74±3.95, VFA 52.00±24.04), normal BMI group (PCOS-NB, n=8, age 27.88±2.53, BMI 21.56±2.20, VFA 27.00±21.18), with no statistical difference in age (P>0.05) and significantly statistical differences in BMI and VFA (P<0.05). The groups showed a significant difference in microbial ß-diversity between PCOS-VO and PCOS-NVO (P=0.002) and no difference between PCOS-NVO and PCOS-NB (P=0.177). Bacteroidetes was the phylum with the highest relative abundance among all patients, followed by Firmicutes. Those with visceral obesity had a higher abundance of Prevotella, Megamonas, and Dialister genera, positively correlated with metabolic markers (r>0.4, P<0.05), and lower abundance of Phascolarctobacterium and Neisseria genera, negatively correlated with metabolic markers (r<-0.4, P<0.05). Functional annotation analysis showed significant differences in relative abundance of ribosome pathway, fatty acid biosynthesis pathway, and sphingolipid signaling pathway between groups, affecting lipid homeostasis and visceral fat accumulation. Conclusion: Alteration in ß-diversity of gut microbiota exists in PCOS with visceral obesity versus those without visceral obesity and relates to functional differences in ribosomes, fatty acid biosynthesis, and sphingolipid signaling pathways.

Identification, biogenesis, and function prediction of a novel circRNA_3238 of chicken.

With the development of high-throughput sequencing, circular RNA has come into people's vision and attracted more and more attention. Many studies have found that circular RNA plays an important role in a variety of biological processes and the occurrence and development of diseases. According to the previous sequencing results, circRNA_3238 was differentially expressed in ALV-J infected group and the non-infected group was selected for subsequent verification and analysis. We found that circRNA_3238 is a stable, circular transcript, which mainly exists in the cytoplasm. And it is widely expressed in various tissues of chickens, and highly expressed in lung, lymph, and bursa of fabricius. Bioinformatics results show that circRNA_3238 and the predicted target genes enriched MAPK signaling pathway, Notch signaling pathway, and other pathways related to disease or immune, revealing circRNA_3238 may indirectly regulate the process of ALV-J infection by regulating target genes.

Prevalence, risk factors, and clinical outcomes of remote intracerebral hemorrhage after intravenous thrombolysis in acute ischemic stroke: a systematic review and meta-analysis.

BACKGROUND: Hemorrhagic transformation (HT) is a common complication of alteplase treatment. However, the prevalence rate, risk factors, and clinical outcomes of remote intracerebral hemorrhage (rICH) after intravenous thrombolysis in acute ischemic stroke are not well understood. METHODS: Following a previously registered protocol, the PubMed, Web of Science, and Embase databases were systematically searched to identify relevant literature up to June 2022. Cohort studies reporting thrombolysis-related rICH in patients with acute ischemic stroke were included. Random effects models were used to calculate pooled prevalence rate, mean difference (MD) or odds ratio (OR) with corresponding 95% confidence interval (CI). RESULTS: Fourteen studies with 52,610 patients were included in this meta-analysis. The pooled rICH prevalence was 3.2% (95% CI 3.1-3.4%). Compared to patients without intracerebral hemorrhage (ICH), those with rICH were older, more likely to be female, and had a higher proportion of prior stroke, chronic heart failure and cardioembolism, and higher diastolic blood pressure. Small vessel disease markers (e.g., white matter hyperintensities and cerebral microbleeds) were strongly associated with rICH. The presence of rICH decreased the likelihood of favorable outcomes (OR 0.36, 95% CI 0.31-0.41) and increased the risk of mortality (OR 4.37, 95% CI 2.86-6.67). CONCLUSIONS: Although rICH is uncommon after intravenous thrombolysis, its presence can lead to worse functional outcomes and higher mortality in acute ischemic stroke. Patients at high risk of rICH must be identified based on potential risk factors.

The 5-Aminolevulinic Acid (5-ALA) Supplement Enhances PSII Photochemical Activity and Antioxidant Activity in the Late Growth Promotion of Pseudostellaria heterophylla.

This study focused on the physiological regulation and mechanism of exogenous 5-aminolevulinic acid (5-ALA) in the late growth of P. heterophylla. In the middle of May, different concentrations of 5-ALA (0, 10, 20, 50 mg·L-1) were sprayed on the leaves. The effects of 5-ALA on tuberous root growth, antioxidant enzyme system, gas exchange, photosynthetic pigment contents and photosynthetic characteristics were measured from 23 May to 13 June. A concentration of 20 mg·L-1 of 5-ALA led to a significant increase in the yield of fresh root and biomass allocation at 38.12% and 25.07%, respectively, in comparation with the control (0 mg·L-1). The moderate concentration of 5-ALA statistically stimulated antioxidation activities. 5-ALA treatment enhanced photosynthetic activity and reduced photodamage. Compared to the control, there were increases in the chlorophyll fluorescence parameters of P. heterophylla under 5-ALA treatment. Moreover, 20 mg·L-1 of 5-ALA significantly changed the kinetic parameters of fluorescence. It enhanced the light absorption and distribution efficiency of PSII and the activities of leaves, resulting in alleviating photoinhibition by the excess excitation energy. The correlation indicated that there was a significant positive correlation between the yield of tuberous roots and biomass allocation, Pn and catalase (CAT), and a negative correlation between the yield of tuberous roots and malondialdehyde (MDA). The appropriate 5-ALA concentration in the late growth stage of P. heterophylla effectively enhanced the net photosynthetic capacity, mainly resulting from the enhancement of PSII photochemical activity to promote the increases in excitation energy absorption, capture and electron transfer efficiency of the leaves. Finally, 5-ALA treatment can increase the photochemical activity of PSII in the whole leaf and ultimately delay the senescence of P. heterophylla.

Selection of fertilization strategies for different sperm parameters in vitro fertilization.

Background: Fertilization is a prerequisite for successful human reproduction. The choice of clinical fertilization strategy is crucial and directly affects clinical outcomes. This study analyzes the most appropriate assisted reproductive technology (ART) strategy based on sperm parameters. Methods: Semen samples were divided into six groups based on semen progressive motility (PR) and semen density (SD): HMLD (high motility-low density) (PR ≥32% and sperm density <15×106/mL, n=60), HMID (high motility-intermediate density) (PR ≥32% and 15×106/mL ≤ SD <30×106/mL, n=106), HMHD (high motility-high density) (PR ≥32% and SD ≥30×106/mL, n=1,009), LMLD (low motility-low density) (PR <32% and SD <15×106/mL, n=99), LMID (low motility-intermediate density) (PR <32% and 15×106/mL ≤ SD <30×106/mL, n=77), and LMHD (low motility-high density) (PR <32% and SD ≥30×106/mL, n=164). We analyzed hyaluronic acid binding (HAB) assay and acrosin activity, along with fertilization, embryonic development, and pregnancy outcomes, to demonstrate the correlation of sperm parameters with fertilization function. Results: In the PR <32% groups, the rate of intracytoplasmic sperm injection (ICSI) treatment decreased with increasing sperm concentration. Specifically, approximately 10% of in vitro fertilization (IVF) treatment cycles required a rescue ICSI when sperm PR was <32% accompanied by SD ≥15×106/mL and PR ≥32% accompanied by SD <30×106/mL, which was significantly higher than HMHD group, P<0.001. Sperm acrosin activity and HAB ability were significantly higher in the groups with good sperm parameters, P<0.05. Conclusions: The findings of this study suggest, fertilization ability of sperm is closely related to sperm motility and density. In clinical practice, IVF strategies should be refined based on male sperm parameters.

Maternal anxiety affects embryo implantation via impairing adrenergic receptor signaling in decidual cells.

Recurrent implantation failure (RIF) is defined as the failed pregnancy after good embryo transfer over 3 cycles during in vitro fertilization (IVF).The human endometrium plays a vital role in providing the site for embryo implantation, with several factors implicated in unsatisfactory endometrial receptivity in RIF. Our present results revealed that women with pregnancy loss or infertility have a higher serum epinephrine level, indicating a potential correlation between psychological stress and pregnancy failure. RNA-sequencing of the tissues collected at the endometrial receptive phase in normal and RIF women showed that stress hormones could affect the functional status of endometrial receptivity. Subsequent analysis revealed that the epinephrine signaling acts as an important regulator of endometrial receptivity through the PI3K-AKT and FOXO1 signaling pathways. We also found that patients with RIF show attenuated expression of the alpha-2C-adrenergic receptor (ADRA2C) and that its down regulation induced by high level epinephrine could inhibit the decidualization. Early pregnant mice treated with stress showed high serum epinephrine levels, defective uterine adrenergic receptor expression, and low pregnancy rates. Altogether, our findings indicate that mental stress during early pregnancy can alter the functional status of endometrial receptivity.

Identification, biogenesis and function prediction of novel circRNA during the chicken ALV-J infection.

Circular RNA (circRNA) is a new non-coding RNA with a highly conserved and stable covalently closed loop structure, and it plays an important role in a variety of biological processes and the occurrence of diseases. Based on the sequencing results, circRNA_3079 had the most significant difference between the infected group and normal group, up to about 8 times. It has attracted our attention and was selected for further verification and analysis. Though the characteristics analysis of circRNA_3079 in chicken, we found circRNA_3079 is a stable, circular transcript, which mainly exists in the cytoplasm. And it is widely expressed in various tissues of chickens, and highly expressed in lung, spleen, lymph and bursa of fabricius. Bioinformatics analysis results showed that circRNA_3079 and the predicted target genes are enriched in many pathways related to immunity or tumors, such as p53 signaling pathway, Jak-STAT signaling pathway and NOD-like receptor signaling pathway, which revealed that circRNA_3079 may indirectly regulate the ALV-J infection process through the regulation of target genes.HIGHLIGHTSCircRNA_3079 is an abundant and stable circular RNA expressed in different tissues and cells in chicken.The circularization of circRNA_3079 does not depend on the reverse complementary repetitive sequence of the flanking sequence.CircRNA_3079 may indirectly regulate the ALV-J infection process.

CircDLGAP4 overexpression relieves oxygen-glucose deprivation-induced neuronal injury by elevating NEGR1 through sponging miR-503-3p.

Circular RNAs (circRNAs) have been reported to play vital regulatory roles in human diseases. However, the functions of circRNAs in ischemic stroke (IS) are limited. In this study, we aimed to explore the functions and mechanisms of circRNA DLG associated protein 4 (circDLGAP4) in IS development. Oxygen-glucose deprivation (OGD)-treated HCN-2 cells were used to mimic IS environment in vitro. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to detect the levels of circDLGAP4, microRNA-503-3p (miR-503-3p) and neuronal growth regulator 1 (NEGR1) mRNA. RNase R assay was conducted to analyze the stability of circDLGAP4. Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis were adopted for cell viability and death, respectively. Western blot assay was performed for protein levels. Enzyme-linked immunosorbent assay (ELISA) kits were used to examine the concentrations of inflammatory cytokines. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull-down assay were employed to analyze the relationships among circDLGAP4, miR-503-3p and NEGR1. CircDLGAP4 level was declined in HCN-2 cells after OGD treatment. CircDLGAP4 overexpression promoted cell viability and suppressed cell death and inflammatory cytokine concentrations in OGD-treated HCN-2 cells. CircDLGAP4 acted as the sponge for miR-503-3p and the impacts of circDLGAP4 overexpression on cell viability, death and inflammation in OGD-treated HCN-2 cells were reversed by miR-503-3p elevation. Furthermore, NEGR1 was the target gene of miR-503-3p. MiR-503-3p inhibition ameliorated OGD-induced HCN-2 cell impairments, but NEGR1 knockdown abolished the effects. CircDLGAP4 alleviated OGD-induced HCN-2 cell damage by regulating miR-503-3p/NEGR1 axis.

Clinical effectiveness of orthodontic miniscrew implantation guided by a novel cone beam CT image-based computer aided design and computer aided manufacturing (CAD-CAM) template.

BACKGROUND: Although miniscrews are widely used in orthodontic treatment as temporary anchorage devices, their correct and safe placement has attracted little attention. This study aimed to introduce a novel cone beam CT (CBCT) image-based computer aided design and computer aided manufacturing (CAD-CAM) template for orthodontic miniscrew implantation and to evaluate the effectiveness of miniscrews implanted under the guidance of this template. METHODS: The CBCT scans of ten patients requiring miniscrews as anchorage were analyzed in NNT software to predetermine the insertion sites of miniscrew implants. The DICOM data of the scans, along with virtual miniscrews acquired in Solidworks software, were imported into Mimics software to construct three-dimensional (3D) images of teeth and bone and to determine the virtual position of miniscrews and 3D virtual templates were designed following consideration of the virtual implantation plans. A STL (Stereolithography) file of the virtual template was output, and the resin template was then fabricated with a stereolithographic appliance (SLA). 24 Miniscrews were then implanted guided by the template and clinical evaluation of their safety and stability, as well as their placement deviations, were made. A dental casts model and cephalometric analysis before and after orthodontic treatment were made to assess the dentomaxillofacial changes. RESULTS: All 24 miniscrews had no contact with adjacent roots. 18 miniscrews had a grade I safety score and six had a grade II. The miniscrews were stable at 1, 3, 6, and 9 months after implantation, although there was mild inflammation around two miniscrews. Implantation deviation of miniscrew in the crown was (1.03±0.65) and (1.26±0.72) mm in the apex, on average. Satisfactory dentomaxillofacial changes in 10 patients with these 24 miniscrews as anchorage were acquired. CONCLUSIONS: Miniscrews could be implanted in the targeted position safely and precisely when guided by the novel templates, and remained stable during orthodontic treatment. Patients treated with these miniscrews as anchorage in orthodontic treatment acquired satisfactory dentomaxillofacial changes.

Cost-effectiveness of second-line nivolumab for platinum-treated advanced non-small-cell lung cancer.

Aim: To analyze the economic impact of nivolumab and chemotherapy in patients with non-small-cell lung cancer (NSCLC) who developed disease progression after platinum-containing dual-drug chemotherapy. Materials & methods: The partitioned survival model was used to analyze the cost-utility of two NSCLC treatments by nivolumab and docetaxel. The clinical data resulted from the Phase III clinical trial. The cost parameters were derived from our previous studies, and the utility parameters were derived from the literature. Results: The quality-adjusted life-years of nivolumab and docetaxel were 0.778 and 0.336. The lifetime direct medical expenses of nivolumab and docetaxel were US$44,707.17 and US$12,826.72. The incremental cost-effectiveness ratio was $72,127.71/quality-adjusted life-year. Conclusion: The combination of chemotherapy, nivolumab is not a cost-effective choice in the second-line treatment of NSCLC.

Expedient Discovery for Novel Antifungal Leads Targeting Succinate Dehydrogenase: Pyrazole-4-formylhydrazide Derivatives Bearing a Diphenyl Ether Fragment.

The pyrazole-4-carboxamide scaffold containing a flexible amide chain has emerged as the molecular skeleton of highly efficient agricultural fungicides targeting succinate dehydrogenase (SDH). Based on the above vital structural features of succinate dehydrogenase inhibitors (SDHI), three types of novel pyrazole-4-formylhydrazine derivatives bearing a diphenyl ether moiety were rationally conceived under the guidance of a virtual docking comparison between bioactive molecules and SDH. Consistent with the virtual verification results of a molecular docking comparison, the in vitro antifungal bioassays indicated that the skeleton structure of title compounds should be optimized as an N'-(4-phenoxyphenyl)-1H-pyrazole-4-carbohydrazide scaffold. Strikingly, N'-(4-phenoxyphenyl)-1H-pyrazole-4-carbohydrazide derivatives 11o against Rhizoctonia solani, 11m against Fusarium graminearum, and 11g against Botrytis cinerea exhibited excellent antifungal effects, with corresponding EC50 values of 0.14, 0.27, and 0.52 µg/mL, which were obviously better than carbendazim against R. solani (0.34 µg/mL) and F. graminearum (0.57 µg/mL) as well as penthiopyrad against B. cinerea (0.83 µg/mL). The relative studies on an in vivo bioassay against R. solani, bioactive evaluation against SDH, and molecular docking were further explored to ascertain the practical value of compound 11o as a potential fungicide targeting SDH. The present work provided a non-negligible complement for the structural optimization of antifungal leads targeting SDH.

Impact of adult sons' marriage squeeze on the mental health of parents in rural China / Impacto do casamento dos filhos adultos na saúde mental dos pais na China rural

ABSTRACT: Studying the influencing factors and interventions of mental health in rural China has important practical significance for promoting personal development and social progress. Using the 2016 waves of the China Family Panel Studies data, this paper examined the effect of adult sons' marriage squeeze on parents' mental health in rural China and its corresponding mechanisms. Our study showed a significant negative association between adult sons' marriage squeeze and the mental health status of rural parents. Moreover, the negative association of adult sons' marriage squeeze with mental health is larger for female parents, parents who are less than 60 years old and parents living with children. In addition, we investigated possible mechanisms of the effect, including intergenerational support, intergenerational relationship and neighborhood relationship, and that adult sons' marriage squeeze mainly damages the mental health of rural parents by increasing the downstream intergenerational support of rural parents and worsening intergenerational and neighborhood relationships.

RESUMO: O estudo dos fatores e intervenções de saúde mental na China rural tem um significado prático importante para promover o desenvolvimento pessoal e o progresso social. Usando dados de 2016 provenientes do China Family Panel Studies (CFPS), este artigo estudou o efeito do "aperto do casamento" de um filho adulto, na saúde mental dos pais, na zona rural da China e seus mecanismos correspondentes. O termo aperto do casamento refere-se a um desequilíbrio entre o número de homens e mulheres disponíveis para se casar em uma determinada sociedade. Nesse estudo evidencia-se uma associação significativamente negativa entre o "aperto no casamento" do filho adulto e o estado de saúde mental dos pais rurais. Além disso, a associação negativa do "aperto do casamento" de um filho adulto com a saúde mental é maior para mães, pais com menos de 60 anos e pais que vivem com filhos. Além disso, investigamos possíveis efeitos, incluindo apoio intergeracional, relacionamento intergeracional e relacionamento de vizinhança, e descobrimos que o aperto do casamento do filho adulto prejudica principalmente a saúde mental dos pais rurais, aumentando o apoio intergeracional e piorando relações intergeracionais e de vizinhança.