Effects of elevated levels of intracellular nitric oxide on Pseudomonas aeruginosa biofilm in chronic skin wound and slow-killing infection models

Nitric oxide (NO), produced through the denitrification pathway, regulates biofilm dynamics through the quorum sensing system in Pseudomonas aeruginosa. NO stimulates P. aeruginosa biofilm dispersal by enhancing phosphodiesterase activity to decrease cyclic di-GMP levels. In a chronic skin wound model containing a mature biofilm, the gene expression of nirS, encoding nitrite reductase to produce NO, was low, leading to reduced intracellular NO levels. Although low-dose NO induces biofilm dispersion, it is unknown whether it influences the formation of P. aeruginosa biofilms in chronic skin wounds. In this study, a P. aeruginosa PAO1 strain with overexpressed nirS was established to investigate NO effects on P. aeruginosa biofilm formation in an ex vivo chronic skin wound model and unravel the underlying molecular mechanisms. Elevated intracellular NO levels altered the biofilm structure in the wound model by inhibiting the expression of quorum sensing–related genes, which was different from an in vitro model. In Caenorhabditis elegans as a slow-killing infection model, elevated intracellular NO levels increased worms’ lifespan by 18%. Worms that fed on the nirS-overexpressed PAO1 strain for 4 h had complete tissue, whereas worms that fed on empty plasmid–containing PAO1 had biofilms on their body, causing severe damage to the head and tail. Thus, elevated intracellular NO levels can inhibit P. aeruginosa biofilm growth in chronic skin wounds and reduce pathogenicity to the host. Targeting NO is a potential approach to control biofilm growth in chronic skin wounds wherein P. aeruginosa biofilms are a persistent problem. (AU)

A deep-learning-based framework for identifying and localizing multiple abnormalities and assessing cardiomegaly in chest X-ray.

Accurate identification and localization of multiple abnormalities are crucial steps in the interpretation of chest X-rays (CXRs); however, the lack of a large CXR dataset with bounding boxes severely constrains accurate localization research based on deep learning. We created a large CXR dataset named CXR-AL14, containing 165,988 CXRs and 253,844 bounding boxes. On the basis of this dataset, a deep-learning-based framework was developed to identify and localize 14 common abnormalities and calculate the cardiothoracic ratio (CTR) simultaneously. The mean average precision values obtained by the model for 14 abnormalities reached 0.572-0.631 with an intersection-over-union threshold of 0.5, and the intraclass correlation coefficient of the CTR algorithm exceeded 0.95 on the held-out, multicentre and prospective test datasets. This framework shows an excellent performance, good generalization ability and strong clinical applicability, which is superior to senior radiologists and suitable for routine clinical settings.

Climate warming worsens thermal resource utilization for practical rice cultivation in China.

Rice production is sensitive to climate change and significantly affected by warming in recent years. To what extent climate warming shifted rice phenology and varied thermal resource condition were explored across five agro-ecological zones in China, based on up-to-date observations of meteorology and rice cultivation in 1981-2020. It was clearly signaled that there was a general advance of 0.3-3.8 days in observed sowing date and a delay of 0.4-3.5 days in observed maturity date in 2001-2020 relative to 1981-2000 in major zones. A vacant time slice of 2.6-28.8 days between observed sowing date and potential sowing date, and a lag of 15.4-56.7 days in potential maturity date compared to observed maturity date were identified in 2001-2020. Within longer growing season, useful accumulated temperature increased by 76.7-117.6 °C·d in 2001-2020 relative to 1981-2000, while disactive accumulated temperature also increased. In Northeast China, actual rice cultivation was undergoing earlier sowing date than potential sowing date and longer growing duration than potential duration, yet leading to upward disactive accumulated temperature. The decrease in the thermal resource utilization in 2001-2020 relative to 1981-2000 was highlighted at 55.3-78.3% stations in major zones, with a decrement of 0.006-0.018 in average magnitude. The changes in thermal resource utilization unveiled that the shifts in actual rice cultivation still could not compensate for the suitability in thermal resource utilization benefited from climate warming.

Relationship between LMAN2 expression in HR-positive breast cancer tissues and patient prognosis and its effect on proliferation and migration of MCF-7 cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究甘露糖结合凝集素2（LMAN2）在激素受体（HR）阳性乳腺癌组织中的表达水平与乳腺癌患者预后的关系及其对MCF-7 细胞增殖和迁移的影响。方法：通过TCGA、Bc-GenExMiner、GEPIA和Kaplan-Meier Plotter数据库分析LMAN2在乳腺癌组织和正常乳腺组织中的差异性表达及其与患者预后的关系。采用小RNA干扰技术将si-LMAN2#1、si-LMAN2#2及si-NC转染至MCF-7细胞，将过表达LMAN载体（pc-LMAN）及空载体pcDNA3.1阴性对照（pc-NC）转染至MCF-7细胞，实验分为si-LMAN2#1、si-LMAN2#2、si-NC、pc-LMAN2和pc-NC组。通过qPCR和WB实验检测各组细胞中LMAN2 mRNA和蛋白的表达水平，CCK-8、克隆形成、Transwell迁移、WB等实验检测敲低和过表达LMAN 2对MCF-7细胞增殖、克隆形成、迁移及AKT信号通路相关蛋白表达的影响。结果：LMAN2在乳腺癌组织中的表达水平显著高于正常乳腺组织（P<0.001）。HR阳性乳腺癌组织中LMAN2表达水平显著高于HR阴性乳腺癌组织（P<0.001）；LMAN2高表达与HR阳性乳腺癌患者不良预后有关联。敲低LMAN2可显著降低MCF-7细胞的增殖和迁移能力（P<0.01或P<0.001），过表达LMAN2可显著提高MCF-7细胞的增殖和迁移能力（均P<0.001）。敲低LMAN2组MCF-7细胞中PTEN和P21蛋白表达水平均显著升高，p-AKT蛋白表达水平显著降低（均P<0.01）。结论：LMAN2在乳腺癌组织和HR阳性乳腺癌组织中高表达，且与不良预后有关联。LMAN2高表达与MCF-7细胞增殖和迁移有关联，其作用机制可能涉及AKT信号通路。

TREX2 enables efficient genome disruption mediated by paired CRISPR-Cas9 nickases that generate 3'-overhanging ends.

Paired SpCas9 nickases (SpCas9n) are an effective strategy to reduce off-target effect in genome editing. However, this approach is not efficient with 3'-overhanging ends, limiting its applications. In order to expand the utility of paired SpCas9n in genome editing, we tested the effect of the TREX2 3'-5' exonuclease on repair of 3'-overhanging ends. We found ectopic overexpression of Trex2 stimulates the efficiency of paired SpCas9n in genome disruption with 3'-overhanging ends up to 400-fold with little stimulation of off-target editing. TREX2 overexpressed preferentially deletes entire 3' overhangs but has no significant effect on 5' overhangs. Trex2 overexpression also stimulates genome disruption by paired SpCas9n that potentially generate short 3'-overhanging ends at overlapping SpCas9n target sites, suggesting sequential nicking of overlapping target sites by SpCas9n. This approach is further simplified with improved efficiency and safety by fusion of TREX2 and particularly its DNA-binding-deficient mutant to SpCas9n. Junction analysis at overlapping targets revealed the different extent of end resection of 3' single-stranded DNA (ssDNA) by free TREX2 and TREX2 fused to SpCas9n. SpCas9n-TREX2 fusion is more convenient and safer than overexpression of free TREX2 to process 3'-overhanging ends for efficient genome disruption by paired SpCas9n, allowing practical use of this TREX2-based strategy in genome editing.

Assessment of key parameters of normal uterus in women of reproductive age.

Currently, the precise and detailed anatomical data of the normal uterus, especially the myometrium thickness in various parts of the uterus, are lacking. This study aims to provide normal references for uterine size in healthy reproductive-aged Chinese women to facilitate the application of hysteroscopic surgery. A total of 298 women of reproductive age with normal uterine were included. Parity was significantly correlated with uterine measurements (P < 0.05), and age impacted several measurements (P < 0.05). At each uterine site examined, the myometrium was thinner in nulliparous women than in parous or primiparous women (P < 0.001). Similarly, the extrauterine measurements for parous or primiparous women were larger than those for nulliparous women. Weight affected some external measurements but not myometrial thicknesses, while height did not affect uterine measurements (P > 0.05). There was a positive correlation between body mass index (BMI) and extrauterine measurements as well as myometrial thickness (P < 0.05). The mathematical model of the uterine size for women of reproductive age was constructed stratified by parity. The study is the first to provide a detailed statistical description of the accurate anatomical parameters of the uterus in Chinese reproductive-aged women and has great significance for improving the safety and effectiveness of hysteroscopic surgery for patients.

Vulvar squamous intraepithelial neoplasia epithelial thickness in hairy and non-hairy sites: a single center experience from China.

Introduction: A large-sample study focusing on VIN lesions of a more precise thickness is needed to help guide clinical treatment. This study aimed to investigate the depth of vulvar intraepithelial neoplasia (VIN) and involved skin appendages to provide evidence for laser surgery. Methods: The study retrospectively enrolled and analyzed the clinical characteristics of VIN patients in the obstetrics and gynecology department of a university hospital between January 1, 2019 and December 30, 2021. The study further explored the thickness of epithelium and skin appendages of 285 women with low-grade VIN (VIN1) and 285 women with high-grade VIN (VIN2/3). Results: The study included 1,139 (80%) VIN1 and 335 (20%) VIN2/3 cases. The VIN1 and VIN2/3 groups showed a significant difference in human papillomavirus infection (P<0.01) but not in cytology (P = 0.499). Most (89.90%, 1,325) cases occurred in one area of the vulva, whereas 10.11% were multifocal. VIN commonly occurred on the posterior fourchette (76.85%), labia majora (11.61%), and labia minora (9.92%). The VIN2/3 group reported a significantly higher positive rate for concurrent cervical and vaginal intraepithelial neoplasia (160 of 285) than the VIN1 group (321 of 953) (P=0.000). The involved epithelial thicknesses in VIN2/3 and VIN1 were 0.69 ± 0.44 and 0.49 ± 0.23 mm, respectively, both of which were greater than the corresponding noninvolved epithelial thickness (0.31 ± 0.19 and 0.32 ± 0.10 mm, P<0.001 and P<0.001, respectively). In cases of appendage involvement, the VIN thickness was 1.98 ± 0.64 mm. Conclusions: VIN thickness was generally ≤1 mm for the superficial lesions in non-hairy areas. However, for lesions extending onto hairy areas, the thickness was approximately 3 mm, leading to the destruction of involved skin appendages.

Alkaline phosphatase LapA regulates quorum sensing-mediated virulence and biofilm formation in Pseudomonas aeruginosa PAO1 under phosphate depletion stress.

IMPORTANCE: Our previous study demonstrated that the expression of lapA was induced under phosphate depletion conditions, but its roles in virulence and biofilm formation by Pseudomonas aeruginosa remain largely unknown. This study presents a systematic investigation of the roles of lapA in virulence induction and biofilm formation by constructing a lapA-deficient strain with P. aeruginosa PAO1. The results showed that deletion of the lapA gene evidently reduced elastase activity, swimming motility, C4-HSL, and 3-oxo-C12-HSL production, and increased rhamnolipid production under phosphate depletion stress. Moreover, lapA gene deletion inhibited PAO1 biofilm formation in porcine skin explants by reducing the expression levels of las and rhl quorum sensing systems and extracellular polymeric substance synthesis. Finally, lapA gene deletion also reduced the virulence of PAO1 in Caenorhabditis elegans in fast-kill and slow-kill infection assays. This study provides insights into the roles of lapA in modulating P. aeruginosa virulence and biofilm formation under phosphate depletion stress.

Neoadjuvant Chemotherapy of Taxanes With or Without Anthracyclines in Different Molecular Subtypes of Breast Cancer: A Propensity Score Matching Study.

PURPOSE: To compare the efficacy of taxane (T) based neoadjuvant chemotherapy (NAC) with T and anthracycline (A) based NAC in different molecular types of breast cancer (BC). METHODS: We retrospectively analyzed the date of NAC for BC from 20 hospitals in China from January 2010 to December 2020, 7870 cases were enrolled. The propensity score matching was used to equalize the baseline characteristics. Pathological complete response (pCR) rate, clinical response rate and breast-conserving rate were analyzed. RESULTS: The efficacy of 2 regimens were similar in luminal A subtype. The breast-conserving rate was higher in T-based NAC in luminal B subtype (17.9% vs. 10.2%, P = .043).The pCR (T0/isN0M0) and tpCR (T0N0M0) rates in T-based NAC were higher than those in TA-based NAC for triple-negative subtype (pCR: 34.5% vs. 25.8%, P = .041, tpCR: 26.9% vs. 17.1%, P = .008). For HER2+(HR-) subtype, the pCR, and tpCR rates were higher in T-based NAC in insufficient anti-HER2 therapy (P < .05), and those were higher in TA-based NAC in dual-target anti-HER2 therapy (pCR: 69.2% vs. 53.8%, P = .254, tpCR: 61.5% vs. 42.3%, P = .165). For HER2+(HR+) breast cancer, both pCR and tpCR rates were higher in TA group, regardless of the adequacy of anti-HER2 treatment. CONCLUSIONS: T-based NAC could replace TA-based NAC for luminal A, luminal B, and triple-negative early-stage BC, but anthracyclines cannot be abandoned in HER2+ breast cancer. The development of anthracyclines with lower adverse reactions is one of the directions for the treatment of HER2+ breast cancer.

Design, strategies, and therapeutics in nanoparticle-based siRNA delivery systems for breast cancer.

Utilizing small interfering RNA (siRNA) as a treatment for cancer, a disease largely driven by genetic aberrations, shows great promise. However, implementing siRNA therapy in clinical practice is challenging due to its limited bioavailability following systemic administration. An attractive approach to address this issue is the use of a nanoparticle (NP) delivery platform, which protects siRNA and delivers it to the cytoplasm of target cells. We provide an overview of design considerations for using lipid-based NPs, polymer-based NPs, and inorganic NPs to improve the efficacy and safety of siRNA delivery. We focus on the chemical structure modification of carriers and NP formulation optimization, NP surface modifications to target breast cancer cells, and the linking strategy and intracellular release of siRNA. As a practical example, recent advances in the development of siRNA therapeutics for treating breast cancer are discussed, with a focus on inhibiting cancer growth, overcoming drug resistance, inhibiting metastasis, and enhancing immunotherapy.

Effects of elevated levels of intracellular nitric oxide on Pseudomonas aeruginosa biofilm in chronic skin wound and slow-killing infection models.

Nitric oxide (NO), produced through the denitrification pathway, regulates biofilm dynamics through the quorum sensing system in Pseudomonas aeruginosa. NO stimulates P. aeruginosa biofilm dispersal by enhancing phosphodiesterase activity to decrease cyclic di-GMP levels. In a chronic skin wound model containing a mature biofilm, the gene expression of nirS, encoding nitrite reductase to produce NO, was low, leading to reduced intracellular NO levels. Although low-dose NO induces biofilm dispersion, it is unknown whether it influences the formation of P. aeruginosa biofilms in chronic skin wounds. In this study, a P. aeruginosa PAO1 strain with overexpressed nirS was established to investigate NO effects on P. aeruginosa biofilm formation in an ex vivo chronic skin wound model and unravel the underlying molecular mechanisms. Elevated intracellular NO levels altered the biofilm structure in the wound model by inhibiting the expression of quorum sensing-related genes, which was different from an in vitro model. In Caenorhabditis elegans as a slow-killing infection model, elevated intracellular NO levels increased worms' lifespan by 18%. Worms that fed on the nirS-overexpressed PAO1 strain for 4 h had complete tissue, whereas worms that fed on empty plasmid-containing PAO1 had biofilms on their body, causing severe damage to the head and tail. Thus, elevated intracellular NO levels can inhibit P. aeruginosa biofilm growth in chronic skin wounds and reduce pathogenicity to the host. Targeting NO is a potential approach to control biofilm growth in chronic skin wounds wherein P. aeruginosa biofilms are a persistent problem.

Three-dimensional High-Resolution Dark-Blood Late Gadolinium Enhancement Imaging for Improved Atrial Scar Evaluation.

Background Radiofrequency ablation (RFA) is a widely used treatment for atrial fibrillation, reducing the risk of cardiac arrhythmia. Detailed visualization and quantification of atrial scarring has the potential to improve preprocedural decision-making and postprocedural prognosis. Conventional bright-blood late gadolinium enhancement (LGE) MRI can help detect atrial scars; however, its suboptimal myocardium to blood contrast inhibits accurate scar estimation. Purpose To develop and test a free-breathing LGE cardiac MRI approach that simultaneously provides high-spatial-resolution dark-blood and bright-blood images for improved atrial scar detection and quantification. Materials and Methods A free-breathing, independent navigator-gated, dark-blood phase-sensitive inversion recovery (PSIR) sequence with whole-heart coverage was developed. Two coregistered high-spatial-resolution (1.25 × 1.25 × 3 mm3) three-dimensional (3D) volumes were acquired in an interleaved manner. The first volume combined inversion recovery and T2 preparation to achieve dark-blood imaging. The second volume functioned as the reference for phase-sensitive reconstruction with built-in T2 preparation for improved bright-blood contrast. The proposed sequence was tested in prospectively enrolled participants who had undergone RFA for atrial fibrillation (mean time since RFA, 89 days ± 26 [SD]) from October 2019 to October 2021. Image contrast was compared with conventional 3D bright-blood PSIR images using the relative signal intensity difference. Furthermore, native scar area quantification obtained from both imaging approaches was compared with measurements obtained with electroanatomic mapping (EAM) as the reference standard. Results A total of 20 participants (mean age, 62 years ± 9; 16 male) who underwent RFA for atrial fibrillation were included. The proposed PSIR sequence successfully acquired 3D high-spatial-resolution volumes in all participants, with a mean scan time of 8.3 minutes ± 2.4. The developed PSIR sequence improved scar to blood contrast compared with conventional PSIR sequence (mean contrast, 0.60 arbitrary units [au] ± 0.18 vs 0.20 au ± 0.19, respectively; P < .01) and correlated with EAM regarding scar area quantification (r = 0.66 [P < .01] vs r = 0.13 [P = .63]). Conclusion In participants who had undergone RFA for atrial fibrillation, an independent navigator-gated dark-blood PSIR sequence produced high-spatial-resolution dark-blood and bright-blood images with improved image contrast and native scar quantification compared with conventional bright-blood images. © RSNA, 2023 Supplemental material is available for this article.

What Matters in Radiological Image Segmentation? Effect of Segmentation Errors on the Diagnostic Related Features.

Segmentation is a crucial step in extracting the medical image features for clinical diagnosis. Though multiple metrics have been proposed to evaluate the segmentation performance, there is no clear study on how or to what extent the segmentation errors will affect the diagnostic related features used in clinical practice. Therefore, we proposed a segmentation robustness plot (SRP) to build the link between segmentation errors and clinical acceptance, where relative area under the curve (R-AUC) was designed to help clinicians to identify the robust diagnostic related image features. In experiments, we first selected representative radiological series from time series (cardiac first-pass perfusion) and spatial series (T2 weighted images on brain tumors) of magnetic resonance images, respectively. Then, dice similarity coefficient (DSC) and Hausdorff distance (HD), as the widely used evaluation metrics, were used to systematically control the degree of the segmentation errors. Finally, the differences between diagnostic related image features extracted from the ground truth and the derived segmentation were analyzed, using the statistical method large sample size T-test to calculate the corresponding p values. The results are denoted in the SRP, where the x-axis indicates the segmentation performance using the aforementioned evaluation metric, and the y-axis shows the severity of the corresponding feature changes, which are expressed in either the p values for a single case or the proportion of patients without significant change. The experimental results in SRP show that when DSC is above 0.95 and HD is below 3 mm, the segmentation errors will not change the features significantly in most cases. However, when segmentation gets worse, additional metrics are required for further analysis. In this way, the proposed SRP indicates the impact of the segmentation errors on the severity of the corresponding feature changes. By using SRP, one could easily define the acceptable segmentation errors in a challenge. Additionally, the R-AUC calculated from SRP provides an objective reference to help the selection of reliable features in image analysis.

Construction and validation of a deterioration model for elderly COVID-19 Sub-variant BA.2 patients.

Rationale: COVID-19 pandemic has imposed tremendous stress and burden on the economy and society worldwide. There is an urgent demand to find a new model to estimate the deterioration of patients inflicted by Omicron variants. Objective: This study aims to develop a model to predict the deterioration of elderly patients inflicted by Omicron Sub-variant BA.2. Methods: COVID-19 patients were randomly divided into the training and the validation cohorts. Both Lasso and Logistic regression analyses were performed to identify prediction factors, which were then selected to build a deterioration model in the training cohort. This model was validated in the validation cohort. Measurements and main results: The deterioration model of COVID-19 was constructed with five indices, including C-reactive protein, neutrophil count/lymphocyte count (NLR), albumin/globulin ratio (A/G), international normalized ratio (INR), and blood urea nitrogen (BUN). The area under the ROC curve (AUC) showed that this model displayed a high accuracy in predicting deterioration, which was 0.85 in the training cohort and 0.85 in the validation cohort. The nomogram provided an easy way to calculate the possibility of deterioration, and the decision curve analysis (DCA) and clinical impact curve analysis (CICA)showed good clinical net profit using this model. Conclusion: The model we constructed can identify and predict the risk of deterioration (requirement for ventilatory support or death) in elderly patients and it is clinically practical, which will facilitate medical decision making and allocating medical resources to those with critical conditions.

Construction and evaluation of intrauterine adhesion model in rats by different methods of mechanical injury.

PURPOSE: The study aimed to establish a stable and effective animal model for the experimental study of intrauterine adhesion (IUA) by evaluating various mechanical injury methods. METHODS: A total of 140 female rats were divided into four groups according to the extent and area of endometrial injury: group A (excision area: 2.0 × 0.5 cm2), group B (excision area: 2.0 × 0.25 cm2), group C (endometrial curettage) and group D (sham operation). On the 3rd, 7th, 15th and 30th day after the operation, the tissue samples of each group were collected, and the uterine cavity stenosis and histological changes were recorded by HE and Masson staining. Immunohistochemistry of CD31 was applied to visualize microvessel density (MVD). The pregnancy rate and the number of gestational sacs were used to evaluate the reproductive outcome. RESULTS: The results showed that endometrium injured by small-area endometrial excision or simple curettage could be repaired. The ratio of fibrosis in groups A and B was higher than that in groups C and group D 30 days after modeling (P < 0.001). The number of endometrial glands and MVD in group A was significantly lower than those in groups B, C and D (P < 0.05). The pregnancy rate in group A was 20%, which was lower than that in groups B (33.3%), C (89%) and D (100%) (P < 0.05). CONCLUSION: Full-thickness endometrial excision has a high rate of success in constructing stable and effective IUA models in rats.

miR-9 targeting RUNX1 improves LPS-induced alveolar hypercoagulation and fibrinolysis inhibition through NF-κB inactivation in ARDS.

BACKGROUND: Acute respiratory distress syndrome (ARDS) is a clinical and pathophysiological complex syndrome with high mortality. Alveolar hypercoagulation and fibrinolytic inhibition constitute the core part of the pathophysiology of ARDS. miR-9 (microRNA-9a-5p) plays an important role in the pathogenesis of ARDS, but whether it regulates alveolar pro-coagulation and fibrinolysis inhibition in ARDS remains to be elucidated. We aimed to determine the contributing role of miR-9 on alveolar hypercoagulation and fibrinolysis inhibition in ARDS. METHODS: In the ARDS animal model, we first observed the miR-9 and runt-related transcription factor 1 (RUNX1) expression in lung tissue, the effects of miR-9 on alveolar hypercoagulation and fibrinolytic inhibition in ARDS rats, and the efficacy of miR-9 on acute lung injury. In the cell, alveolar epithelial cells type II (AECII) were treated with LPS, and the levels of miR-9 and RUNX1 were detected. Then we observed the effects of miR-9 on procoagulant and fibrinolysis inhibitor factors in cells. Finally, we explored whether the efficacies of miR-9 were associated with RUNX1; we also preliminarily examined the miR-9 and RUNX1 levels in plasma in patients with ARDS. RESULTS: In ARDS rats, miR-9 expression decreased, but RUNX1 expression increased in the pulmonary tissue of ARDS rats. miR-9 displayed to attenuate lung injury and pulmonary wet/dry ratio. Study results in vivo demonstrated that miR-9 ameliorated alveolar hypercoagulation and fibrinolysis inhibition and attenuated the collagen III expressions in tissue. miR-9 also inhibited NF-κB signaling pathway activation in ARDS. In LPS-induced AECII, the expression changes of both miR-9 and RUNX1 were similar to those in pulmonary tissue in the animal ARDS model. miR-9 effectively inhabited tissue factor (TF), plasma activator inhibitor (PAI-1) expressions, and NF-κB activation in LPS-treated ACEII cells. Besides, miR-9 directly targeted RUNX1, inhibiting TF and PAI-1 expression and attenuating NF-κB activation in LPS-treated AECII cells. Clinically, we preliminarily found that the expression of miR-9 was significantly reduced in ARDS patients compared to non-ARDS patients. CONCLUSION: Our experimental data indicate that by directly targeting RUNX1, miR-9 improves alveolar hypercoagulation and fibrinolysis inhibition via suppressing NF-κB pathway activation in LPS-induced rat ARDS, implying that miR-9/RUNX1 is expected to be a new therapeutic target for ARDS treatment.

C-reactive protein to lymphocyte ratio is a significant predictive factor for poor short-term clinical outcomes of SARS-CoV-2 BA.2.2 patients.

Objective: The aim of the present study is to assess the utility of C-reactive protein to Lymphocyte Ratio (CLR) in predicting short-term clinical outcomes of patients infected by SARS-CoV-2 BA.2.2. Methods: This retrospective study was performed on 1,219 patients with laboratory-confirmed SARS-CoV-2 BA.2.2 to determine the association of CLR with short-term clinical outcomes. Independent Chi square test, Rank sum test, and binary logistic regression analysis were performed to calculate mean differences and adjusted odds ratios (aORs) with their 95% CI, respectively. Results: Over 8% of patients admitted due to SARS-CoV-2 BA.2.2. were critically ill. The best cut-off value of CLR was 21.25 in the ROC with a sensitivity of 72.3% and a specificity of 86%. After adjusting age, gender, and comorbidities, binary logistic regression analysis showed that elevated CLR was an independent risk factor for poor short-term clinical outcomes of COVID-19 patients. Conclusion: C-reactive protein to Lymphocyte Ratio is a significant predictive factor for poor short-term clinical outcomes of SARS-CoV-2 BA.2.2 inflicted patients.

Clinical Diagnostic Value of Atypical Glandular Cells in Cervical Cytology: A Single Center Experience From China.

OBJECTIVE: This study aimed to investigate the diagnostic value of atypical glandular cells (AGCs) by analyzing the prevalence and histopathology of AGCs according to cervical cytology. METHODS: The authors retrospectively reviewed and analyzed the demographic characteristics and histopathological outcomes including pathological diagnosis, pathological site, and epithelial distribution of the AGC cases that were diagnosed by cervical cytology. RESULTS: A total of 387 AGC patients with follow-up records were included. Among them, the prevalence of AGC-not otherwise specified (NOS) and AGC-favor neoplastic (FN) was 73.39% (284/387) and 26.62% (103/387), respectively. The high-risk human papillomavirus (hr-HPV)-positive rate was higher in AGC-FN than in AGC-NOS ( p = .002). The difference in pathological severity was statistically significant between hr-HPV-positive and negative AGC patients ( p = .010). Hr-HPV-positive AGC mainly occurs in cervical diseases, whereas hr-HPV-negative AGC is mainly related to endometrial lesions. Precancerous or malignant lesions were found in 36.43% (141/387) of AGC cases and were more commonly seen in AGC-FN than AGC-NOS ( p < .001). The histopathological severity and the incidence of uterine disease were higher among AGC women aged 40 years and older than those younger than 40 years ( p < .05). The possibility of the abnormal origin of glandular epithelial was higher than that of squamous epithelial in AGC patients aged 40 years and older ( p = .0003). CONCLUSIONS: The management of AGC women by age triage is reasonable because the incidence of the glandular epithelial lesion and uterine disease increases in AGC patients 40 years or older. Standardized clinical diagnosis and regular follow-up are recommended for all AGC patients.

Proximal binding of dCas9 at a DNA double strand break stimulates homology-directed repair as a local inhibitor of classical non-homologous end joining.

In CRISPR/Cas9 genome editing, the tight and persistent target binding of Cas9 provides an opportunity for efficient genetic and epigenetic modification on genome. In particular, technologies based on catalytically dead Cas9 (dCas9) have been developed to enable genomic regulation and live imaging in a site-specific manner. While post-cleavage target residence of CRISPR/Cas9 could alter the pathway choice in repair of Cas9-induced DNA double strand breaks (DSBs), it is possible that dCas9 residing adjacent to a break may also determine the repair pathway for this DSB, providing an opportunity to control genome editing. Here, we found that loading dCas9 onto a DSB-adjacent site stimulated homology-directed repair (HDR) of this DSB by locally blocking recruitment of classical non-homologous end-joining (c-NHEJ) factors and suppressing c-NHEJ in mammalian cells. We further repurposed dCas9 proximal binding to increase HDR-mediated CRISPR genome editing by up to 4-fold while avoiding exacerbation of off-target effects. This dCas9-based local inhibitor provided a novel strategy of c-NHEJ inhibition in CRISPR genome editing in place of small molecule c-NHEJ inhibitors, which are often used to increase HDR-mediated genome editing but undesirably exacerbate off-target effects.