Higher ETCOc predicts longer phototherapy treatment in neonatal hyperbilirubinemia.

Objective: This study aimed to evaluate the predictive performance of end-tidal carbon monoxide corrected to ambient carbon monoxide (ETCOc) values phototherapy in neonates with significant hyperbilirubinemia. Methods: A prospective study was conducted on neonates with significant hyperbilirubinemia who received phototherapy between 3 and 7 days of life. The breath ETCOc and serum total bilirubin of the recruited infants were measured on admission. Results: The mean ETCOc at admission in 103 neonates with significant hyperbilirubinemia was 1.70 ppm. The neonates were categorized into two groups: phototherapy duration ≤72 h (n = 87) and >72 h (n = 16) groups. Infants who received phototherapy for >72 h had significantly higher ETCOc (2.45 vs. 1.60, P = 0.001). The cutoff value of ETCOc on admission for predicting longer phototherapy duration was 2.4 ppm, with a sensitivity of 62.5% and specificity of 88.5%, yielding a 50% positive predictive value and a 92.7% negative predictive value. Conclusion: ETCOc on admission can help predict the duration of phototherapy in neonates with hyperbilirubinemia, facilitate clinicians to judge disease severity, and make clinical communication easier and more efficient.

MCIBI++: Soft Mining Contextual Information Beyond Image for Semantic Segmentation.

Co-occurrent visual pattern makes context aggregation become an essential paradigm for semantic segmentation. The existing studies focus on modeling the contexts within image while neglecting the valuable semantics of the corresponding category beyond image. To this end, we propose a novel soft mining contextual information beyond image paradigm named MCIBI++ to further boost the pixel-level representations. Specifically, we first set up a dynamically updated memory module to store the dataset-level distribution information of various categories and then leverage the information to yield the dataset-level category representations during network forward. After that, we generate a class probability distribution for each pixel representation and conduct the dataset-level context aggregation with the class probability distribution as weights. Finally, the original pixel representations are augmented with the aggregated dataset-level and the conventional image-level contextual information. Moreover, in the inference phase, we additionally design a coarse-to-fine iterative inference strategy to further boost the segmentation results. MCIBI++ can be effortlessly incorporated into the existing segmentation frameworks and bring consistent performance improvements. Also, MCIBI++ can be extended into the video semantic segmentation framework with considerable improvements over the baseline. Equipped with MCIBI++, we achieved the state-of-the-art performance on seven challenging image or video semantic segmentation benchmarks.

Feasibility and basic acoustic characteristics of intelligent long-term bowel sound analysis in term neonates.

Objective: Bowel dysfunction continues to be a serious issue in neonates. Traditional auscultation of bowel sounds as a diagnostic tool in neonatal gastrointestinal problems is limited by skill and inability to document and reassess. Consequently, in order to objectively and noninvasively examine the viability of continuous assessment of bowel sounds, we utilized an acoustic recording and analysis system to capture bowel sounds and extract acoustic features in term neonates. Methods: From May 1, 2020 to September 30, 2020, 82 neonates who were hospitalized because of hyperbilirubinemia were included. For 20 h, a convolutional neural network-based acoustic recorder that offers real-time, wireless, continuous auscultation was employed to track the bowel sounds of these neonates. Results: (1) Usable data on five acoustic parameters of bowel sound was recorded for 68 neonates, and the median values were as follows: The rate was 25.80 times/min [interquartile range (IQR): 15.63-36.20]; the duration was 8.00 s/min (IQR: 4.2-13.20); the amplitude was 0.46 (IQR: 0.27-0.68); the frequency was 944.05 Hz (IQR: 848.78-1,034.90); and the interval time was 2.12 s (IQR: 1.3-3.5). (2) In comparison to the parameters of the bowel sounds recorded from the right lower abdomen in 68 infants, the acoustic parameters of the 10 out of 68 infants from chest controls and blank controls were considerably different. (3) The 50%-75% breast milk intake group had the highest rate, the longest duration, and the highest amplitude of bowel sounds, while the >75% breast milk intake group had the highest frequency of bowel sounds. (4) Compared with neonates without hyperbilirubinemia, there was no significant difference in the five parameters of bowel sounds in hyperbilirubinemia infants; nor was there a significant effect of phototherapy and non-phototherapy status on the parameters of bowel sounds during bowel sound monitoring in hyperbilirubinemia patients. (5) A mild transient skin rash appeared on the skin of three infants. No other adverse events occurred. Conclusion: The acoustic recording and analysis system appears useful for monitoring bowel sounds using a continuous, invasive, and real-time approach. Neonatal bowel sounds are affected by various feeding types rather than hyperbilirubinemia and phototherapy. Potential influencing factors and the significance of their application in neonatal intestinal-related disorders require further research.

Comparisons of care practices for very preterm infants and their short-term outcomes in two tertiary centers in northwest and south China: A retrospective cohort study.

BACKGROUND: Care practices for very preterm infants and the mortality and morbidity of the infants vary widely among countries and regions with different levels of economic development, including the different areas in China. We aimed to compare the obstetric and delivery room practices of two representative tertiary newborn centers in the northwestern and southern regions of China and the mortality and morbidity of their very preterm infants. METHODS: A retrospective cohort study was conducted. Very preterm infants born between 220/7 and 316/7 weeks of gestation, and admitted to Qinghai Red Cross Hospital (QHH) and Shenzhen Baoan Women's and Children's Hospital (SZH) from January 1, 2018 to December 31, 2020, were included. The infants' characteristics and short-term outcomes, and the hospitals' care practices were compared between the two cohorts. RESULTS: Three hundred and two infants in QHH and 505 infants in SZH were enrolled, and the QHH cohort was more mature than the SZH cohort was (gestational age 30.14 (29.14-31.14) vs. 29.86 (27.86-31.00 weeks, respectively), p < 0.001). Fewer antenatal steroids and more tracheal intubations were used in QHH than in SZH [(73.8% vs. 90.9%, p < 0.001) and (68.2% vs. 35.0%, p < 0.001, respectively)]. The odds of mortality [aOR = 10.31, 95%CI: (6.04, 17.61)], mortality or major morbidity [aOR = 5.95, 95%CI: (4.05, 8.74)], mortality despite active treatment [aOR = 3.14, 95%CI: (1.31, 7.53)], mortality or major morbidity despite active treatment [aOR = 3.35, 95%CI: (2.17, 5.17)], moderate or severe bronchopulmonary dysplasia [aOR = 3.66, 95%CI: (2.20, 6.06)], and severe retinopathy of prematurity [aOR = 3.24, 95%CI: (1.19, 8.83)] were higher in the QHH cohort. No significant difference in the rate of severe neurological injury or necrotizing enterocolitis ≥ Stage 2 was found between the cohorts. CONCLUSION: Obstetric and delivery room care practices used in the management of very preterm infants differed considerably between the QHH and SZH cohorts. Very preterm infants born in QHH have higher odds of mortality or severe morbidity compared with those born in SZH.

Prediction Models for Bronchopulmonary Dysplasia in Preterm Infants: A Systematic Review.

Objective: To provide an overview and critical appraisal of prediction models for bronchopulmonary dysplasia (BPD) in preterm infants. Methods: We searched PubMed, Embase, and the Cochrane Library to identify relevant studies (up to November 2021). We included studies that reported prediction model development and/or validation of BPD in preterm infants born at ≤32 weeks and/or ≤1,500 g birth weight. We extracted the data independently based on the CHecklist for critical Appraisal and data extraction for systematic Reviews of prediction Modelling Studies (CHARMS). We assessed risk of bias and applicability independently using the Prediction model Risk Of Bias ASsessment Tool (PROBAST). Results: Twenty-one prediction models from 13 studies reporting on model development and 21 models from 10 studies reporting on external validation were included. Oxygen dependency at 36 weeks' postmenstrual age was the most frequently reported outcome in both development studies (71%) and validation studies (81%). The most frequently used predictors in the models were birth weight (67%), gestational age (62%), and sex (52%). Nearly all included studies had high risk of bias, most often due to inadequate analysis. Small sample sizes and insufficient event patients were common in both study types. Missing data were often not reported or were discarded. Most studies reported on the models' discrimination, while calibration was seldom assessed (development, 19%; validation, 10%). Internal validation was lacking in 69% of development studies. Conclusion: The included studies had many methodological shortcomings. Future work should focus on following the recommended approaches for developing and validating BPD prediction models.

SAFNet: A Semi-anchor-free Network with Enhanced Feature Pyramid for Object Detection.

In recent years, the field of object detection has made significant progress. The success of most of the state-of-the-art object detectors is derived from the use of feature pyramid and the carefully designed anchor boxes. However, the current methods of constructing feature pyramid usually blindly integrate multi-scale representations on each feature hierarchy. Furthermore, these detectors also suffer from some drawbacks brought by the hand-designed anchors. To mitigate the adverse effects caused thereby, we introduce a one-stage object detector, named as the semi-anchor-free network with enhanced feature pyramid (SAFNet). Specifically, to better construct feature pyramid, we propose a novel enhanced feature pyramid generation paradigm, which mainly consists of two modules, i.e., adaptive feature fusion module (AFFM) and self-enhanced module (SEM). The paradigm adaptively integrates multi-scale representations in a non-linear method meanwhile suppress the redundant semantic information for each pyramid level, such that a clean and enhanced feature pyramid could be obtained. In addition, an adaptive anchor generator (AAG) is designed to yield fewer but more suitable anchor boxes for each input image. Benefiting from the enhanced feature pyramid, AAG is capable of generating more accurate anchor boxes by introducing few priors. Thus, AAG has the ability to alleviate the drawbacks caused by the preset anchor hyper-parameters and helps to decrease the computation cost. Extensive experiments demonstrate the effectiveness of our approach. Profited from the proposed modules, SAFNet significantly boosts the detection performance, i.e., achieving 2 points and 2.1 points higher Average Precision (AP) than RetinaNet (our baseline) on PASCAL VOC and MS COCO respectively. Codes will be publicly available soon.

A novel TLR7 agonist as adjuvant to stimulate high quality HBsAg-specific immune responses in an HBV mouse model.

BACKGROUND: The global burden of hepatitis B virus (HBV) infection in terms of morbidity and mortality is immense. Novel treatments that can induce a protective immune response are urgently needed to effectively control the HBV epidemic and eventually eradicate chronic HBV infection. METHODS: We designed and evaluated an HBV therapeutic vaccine consisting of a novel Toll-like receptor 7 (TLR7) agonist T7-EA, an Alum adjuvant and a recombinant HBsAg protein. We used RNA-seq, ELISA and hTLR7/8 reporting assays to characterize T7-EA in vitro and real-time PCR to evaluate the tissue-retention characteristics in vivo. To evaluate the adjuvant potential, we administrated T7-EA intraperitoneally in a formulation with an Alum adjuvant and HBsAg in normal and HBV mice, then, we evaluated the HBsAg-specific immune responses by ELISA and Elispot assays. RESULTS: T7-EA acted as an hTLR7-specific agonist and induced a similar gene expression pattern as an unmodified TLR7 ligand when Raw 264.7 cells were exposed to T7-EA; however, T7-EA was more potent than the unmodified TLR7 ligand. In vivo studies showed that T7-EA had tissue-retaining activity with stimulating local cytokine and chemokine expression for up to 7 days. T7-EA could induce Th1-type immune responses, as evidenced by an increased HBsAg-specific IgG2a titer and a T-cell response in normal mice compared to mice received traditional Alum-adjuvant HBV vaccine. Importantly, T7-EA could break immune tolerance and induce persistent HBsAg-specific antibody and T-cell responses in an HBV mouse model. CONCLUSIONS: T7-EA might be a candidate adjuvant in a prophylactic and therapeutic HBV vaccine.

Perinatal Antibiotic Exposure Affects the Transmission between Maternal and Neonatal Microbiota and Is Associated with Early-Onset Sepsis.

Intrapartum antibiotic prophylaxis reduces the risk of infection to a mother and neonate, but antibiotic-mediated maternal and neonatal microbiota dysbiosis increases other health risks to newborn infants. We studied the impact of perinatal antibiotic prophylaxis on the microbiota in mothers and newborns with full-term or preterm delivery. Ninety-eight pregnant women and their neonates were divided into the following four groups: full term without antibiotic exposure (FT), full term with antibiotic exposure (FTA), preterm without antibiotic exposure (PT), and preterm with antibiotic exposure (PTA). Bacterial composition was analyzed by sequencing the 16S rRNA gene from maternal vaginal swabs (V) and neonatal meconium (F). The results showed that in maternal vaginal and neonatal meconium microbiota, FT and PT groups had a higher load of Lactobacillus spp. than did the FTA and PTA groups. In addition, whether in the mother or newborn, the dissimilarity in microbiota between FT and PT was the lowest compared to that between other groups. Compared to the FT and PT groups, the dissimilarity in microbial structures between the vagina and meconium decreased in the FTA and PTA groups. The health outcome of infants reveals an association between early-onset sepsis and antibiotic-mediated microbiota dysbiosis. In conclusion, perinatal antibiotic exposure is related to the establishment of gut microbiota and health risks in newborns. Promoting the rational usage of antibiotics with pregnant women will improve neonatal health.IMPORTANCE Perinatal antibiotic prophylaxis is an effective method for preventing group B Streptococcus (GBS) infection in newborns. Antibiotic exposure unbalances women's vaginal microbiota, which is associated with the establishment of the newborn gut microbiota. However, the influence of perinatal antibiotic exposure on neonatal gut microbiota colonization and health outcomes remains unclear. In this study, we found that perinatal antibiotic exposure induced microbiota dysbiosis in a woman's vagina and the neonatal gut, and we highlight a significant decrease in the abundance of Lactobacillus spp. The influence of antibiotic use on the microbiota was greater than that from gestational age. Additionally, full-term newborns without antibiotic exposure had no evidence of early-onset sepsis, whereas in full-term or preterm newborns with antibiotic exposure before birth, at least one infant was diagnosed with early-onset sepsis. These results suggest an association between perinatal antibiotic exposure and microbial dysbiosis in maternal vaginal and neonatal gut environments, which may be related to the occurrence of early-onset sepsis.

Suppressive effects of sunitinib on a TLR activation-induced cytokine storm.

The cytokine storm includes a clinically heterogeneous set of life-threatening conditions that are manifested by extremely elevated serum cytokine levels and related symptoms (e.g., septic shock) and is devilishly mediated by Toll-like receptor (TLR) agonists in most situations. A tyrosine kinase inhibitor (TKIs), sunitinib, was screened in our group previously and showed antagonistic activity for cytokine release in a TLR7 stimulation model. In this paper, we further studied its mechanisms on interesting phenomena. In vitro, nearly all of the eleven TKIs decreased the TNF-α levels induced by the TLR7 agonist, especially sunitinib. Furthermore, sunitinib displayed potent inhibition of the cytokine levels triggered by several types of TLR ligands, including TLR3, TLR4, TLR7/8 and TLR9, in mouse spleen lymphocytes, mouse BMDCs and human PBMCs. The in vivo results showed that sunitinib efficiently depressed the LPS-induced cytokine storm, i.e., rapid and intense production of TNF-α and IL-6. Sunitinib further increased the survival time and decreased damage to mice. As for the immunosuppressive mechanisms of sunitinib, at least the PDGFR-activated ERK and p38 pathways were critical, although we could not rule out the possibility of other pathways being involved. In conclusion, our study demonstrated the inhibitory actions of TKIs on the cytokine storm induced by TLR ligands, primarily through PDGFR pathways, which could be potentially used to reduce cytokine storms in septic shock.

Combining thioridazine and loratadine for the treatment of gastrointestinal tumor.

In 2015, the American Society of Clinical Oncology announced that strategies of using combination therapies have been indicated to be effective against many types of cancer. In the present study, thioridazine (THZ) was used in a combination therapy with loratadine (LOR) to target gastrointestinal tumor, with the aim of investigating whether combined therapy was superior to monotherapy in its antitumor effects. The antiproliferative effects on CT26.WT and MFC cells were analyzed using cell-counting kit-8 assay, and synergistic effect was assessed by combination index (Fig. 1). Annexin V and propidium iodide staining indicated the combination therapy was able to induce apoptosis and that this may be mediated via caspase-3, -9 and poly (ADP-ribose) polymerase (PARP) (Fig. 2). Antitumor activity was also evaluated in CT26.WT xenografts in BALB/c mice (Fig. 3). Furthermore, as expected, combination therapy was able to successfully inhibit the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling pathway (Fig. 4). These findings suggest that the combination therapy with THZ and LOR may provide a promising therapy for gastrointestinal cancer.

Immunomodulatory and Antitumor Effects of a Novel TLR7 Agonist Combined with Lapatinib.

As new treatment approaches, both immunotherapy and targeted treatments have been used in the clinical treatment of cancers. These therapies are different from traditional surgery, chemotherapy and radiotherapy. Use of a combination of immunotherapy and targeted treatments may improve tumor clearance. We investigated the feasibility of combining tyrosine kinase inhibitors (TKIs, targeted drugs) and SZU-101 (a novel TLR7 agonist synthesized by our laboratory). Thirteen different TKIs were combined with or without SZU-101 and studied to determine their effects on immunocytes. On the basis of the distinctive results, lapatinib and sunitinib were selected for further tumor-inhibition investigation and determination of the underlying mechanism. Interestingly, we found lapatinib to work better with SZU-101, enhancing tumor clearance in vivo, without affecting the TLR7-NF-κB pathway activated by the TLR7 agonist in mouse spleen lymphocytes and bone marrow dendritic cells (BMDCs).

Ethacrynic acid improves the antitumor effects of irreversible epidermal growth factor receptor tyrosine kinase inhibitors in breast cancer.

Prolonged treatment of breast cancer with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) often results in acquired resistance and a narrow therapeutic index. One strategy to improve the therapeutic effects of EGFR TKIs is to combine them with drugs used for other clinical indications. Ethacrynic acid (EA) is an FDA approved drug that may have antitumor effects and may enhance the cytotoxicity of chemotherapeutic agents by binding to glutathione and inhibiting WNT signaling. While the α,ß-unsaturated-keto structure of EA is similar to that of irreversible TKIs, the mechanism of action of EA when combined with irreversible EGFR TKIs in breast cancer remains unknown. We therefore investigated the combination of irreversible EGFR TKIs and EA. We found that irreversible EGFR TKIs and EA synergistically inhibit breast cancer both in vitro and in vivo. The combination of EGFR TKIs and EA induces necrosis and cell cycle arrest and represses WNT/ß-catenin signaling as well as MAPK-ERK1/2 signaling. We conclude that EA synergistically enhances the antitumor effects of irreversible EGFR TKIs in breast cancer.

[Combination of TLR7 agonist T7-ethacrynic acid conjugate with ROR1 has a stronger anti-breast cancer effect].

Objective To investigate the synergistic anti-breast cancer effect of Toll-like receptor 7 agonist T7-ethacrynic acid conjugate (T7-EA) in combination with receptor-tyrosine-kinase-like orphan receptor 1 (ROR1). Methods ROR1 cytotoxic T lymphocyte (CTL) epitope was predicted using Syfpeithi online software. Mouse spleen lymphocytes and bone marrow dendritic cells (DCs) were separately stimulated with 4 µmol/L T7-EA and 4 µmol/L ROR1 alone or in combination. ELISA assay was used to measure the levels of interferon-γ (IFN-γ), interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α). Xenograft model was established via subcutaneous injection of mouse breast cancer 4T1 cells. The mice were weekly treated through intraperitoneal administration of 3 mg/kg T7-EA, 15 mg/kg ROR1 or the combination of T7-EA and ROR1. After four rounds of treatment, tumor tissues were weighed. Serum level of anti-4T1 tumor protein IgG was measured by ELISA. Specific CTL activity was detected by lactate dehydrogenase (LDH) assay. Results The peptide PYCDETSSV was chosen as an antigen epitope of breast cancer. The T7-EA highly activated in vitro lymphocytes in a dose-dependent manner, which wasn't affected by other relevant peptides. The combination of T7-EA and ROR1 stimulated the secretion of IFN-γ and IL-12 by lymphocytes and TNF-α by bone marrow DCs. The growth of tumor in vivo was significantly inhibited by T7-EA combined with ROR1 compared with T7-EA or ROR1 alone. The specific CTL activity triggered by T7-EA combined with ROR1 was much stronger than that triggered by T7-EA or ROR1 alone. The titer of anti-4T1 tumor protein IgG induced by T7-EA combined with ROR1 was higher than that induced by T7-EA or ROR1. Conclusion The combination of T7-EA and ROR1 has a better killing effect on breast cancer.

Antitumor activity of a novel small molecule TLR7 agonist via immune response induction and tumor microenvironment modulation.

Immunotherapy is emerging as a powerful and active tumor-specific approach against cancer via triggering the immune system. Toll-like receptors (TLRs) are fundamental elements of the immune system, which facilitate our understanding of the innate and adaptive immune pathways. TLR agonists used as single agents can effectively eradicate tumors due to their potent stimulation of innate and adaptive immunity. We examined the effects of a novel adenine type of TLR7 agonists on both innate and adaptive immune activation in vitro and in vivo. We established the local and distant tumor­bearing mice derived from murine mammary carcinoma cell line (4T1) to model metastatic disease. Our data demonstrated that SZU101 was able to stimulate innate immune cells to release cytokines at the very high level compared with LPS at the same or lower concentration. Locally intratumoral SZU101 injection can elicit a systemic antitumor effect on murine breast tumor model. SZU101 affected the frequency of intratumoral immune cell infiltration, including the percentage of CD4+ and CD8+ increase, and the ratio of Tregs decrease. Our data reveal that the antitumor effect of SZU101 is associated with multiple mechanisms, inducing tumor­specific immune response, activation of innate immune cells and modulation of the tumor microenvironment.