Deep learning-assisted diagnosis of benign and malignant parotid tumors based on ultrasound: a retrospective study.

BACKGROUND: To develop a deep learning(DL) model utilizing ultrasound images, and evaluate its efficacy in distinguishing between benign and malignant parotid tumors (PTs), as well as its practicality in assisting clinicians with accurate diagnosis. METHODS: A total of 2211 ultrasound images of 980 pathologically confirmed PTs (Training set: n = 721; Validation set: n = 82; Internal-test set: n = 89; External-test set: n = 88) from 907 patients were retrospectively included in this study. The optimal model was selected and the diagnostic performance evaluation is conducted by utilizing the area under curve (AUC) of the receiver-operating characteristic(ROC) based on five different DL networks constructed at varying depths. Furthermore, a comparison of different seniority radiologists was made in the presence of the optimal auxiliary diagnosis model. Additionally, the diagnostic confusion matrix of the optimal model was calculated, and an analysis and summary of misjudged cases' characteristics were conducted. RESULTS: The Resnet18 demonstrated superior diagnostic performance, with an AUC value of 0.947, accuracy of 88.5%, sensitivity of 78.2%, and specificity of 92.7% in internal-test set, and with an AUC value of 0.925, accuracy of 89.8%, sensitivity of 83.3%, and specificity of 90.6% in external-test set. The PTs were subjectively assessed twice by six radiologists, both with and without the assisted of the model. With the assisted of the model, both junior and senior radiologists demonstrated enhanced diagnostic performance. In the internal-test set, there was an increase in AUC values by 0.062 and 0.082 for junior radiologists respectively, while senior radiologists experienced an improvement of 0.066 and 0.106 in their respective AUC values. CONCLUSIONS: The DL model based on ultrasound images demonstrates exceptional capability in distinguishing between benign and malignant PTs, thereby assisting radiologists of varying expertise levels to achieve heightened diagnostic performance, and serve as a noninvasive imaging adjunct diagnostic method for clinical purposes.

Faecal Microbiota Transplantation Alleviates Ferroptosis after Ischaemic Stroke.

Ischaemic stroke can induce changes in the abundance of gut microbiota constituents, and the outcome of stroke may also be influenced by the gut microbiota. This study aimed to determine whether gut microbiota transplantation could rescue changes in the gut microbiota and reduce ferroptosis after stroke in rats. Male Sprague-Dawley rats (6 weeks of age) were subjected to ischaemic stroke by middle cerebral artery occlusion (MCAO). Fecal samples were collected for 16S ribosomal RNA (rRNA) sequencing to analyze the effects of FMT on the gut microbiota. Neurological deficits were evaluated using the Longa score. triphenyl tetrazolium chloride (TTC) staining was performed in the brain, and kits were used to measure malondialdehyde (MDA), iron, and glutathione (GSH) levels in the ipsilateral brain of rats. Western blotting was used to detect the protein expression levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), and the transferrin receptor 2 (TFR2) in the ipsilateral brain of rats. Stroke induced significant changes in the gut microbiota, and FMT ameliorated these changes. TTC staining results showed that FMT reduced cerebral infarct volume. In addition, FMT diminished MDA and iron levels and elevated GSH levels in the ipsilateral brain. Western blot analysis showed that FMT increased GPX4 and SLC7A11 protein expression and decreased TFR2 protein expression in the ipsilateral brain after stroke. FMT can reverse gut microbiota dysbiosis, reduce cerebellar infarct volume, and decrease ferroptosis after stroke.

Deep learning to assist composition classification and thyroid solid nodule diagnosis: a multicenter diagnostic study.

OBJECTIVES: This study aimed to propose a deep learning (DL)-based framework for identifying the composition of thyroid nodules and assessing their malignancy risk. METHODS: We conducted a retrospective multicenter study using ultrasound images from four hospitals. Convolutional neural network (CNN) models were constructed to classify ultrasound images of thyroid nodules into solid and non-solid, as well as benign and malignant. A total of 11,201 images of 6784 nodules were used for training, validation, and testing. The area under the receiver-operating characteristic curve (AUC) was employed as the primary evaluation index. RESULTS: The models had AUCs higher than 0.91 in the benign and malignant grading of solid thyroid nodules, with the Inception-ResNet AUC being the highest at 0.94. In the test set, the best algorithm for identifying benign and malignant thyroid nodules had a sensitivity of 0.88, and a specificity of 0.86. In the human vs. DL test set, the best algorithm had a sensitivity of 0.93, and a specificity of 0.86. The Inception-ResNet model performed better than the senior physicians (p < 0.001). The sensitivity and specificity of the optimal model based on the external test set were 0.90 and 0.75, respectively. CONCLUSIONS: This research demonstrates that CNNs can assist thyroid nodule diagnosis and reduce the rate of unnecessary fine-needle aspiration (FNA). CLINICAL RELEVANCE STATEMENT: High-resolution ultrasound has led to increased detection of thyroid nodules. This results in unnecessary fine-needle aspiration and anxiety for patients whose nodules are benign. Deep learning can solve these problems to some extent. KEY POINTS: • Thyroid solid nodules have a high probability of malignancy. • Our models can improve the differentiation between benign and malignant solid thyroid nodules. • The differential performance of one model was superior to that of senior radiologists. Applying this could reduce the rate of unnecessary fine-needle aspiration of solid thyroid nodules.

Cost-effectiveness of active surveillance versus early surgery for thyroid micropapillary carcinoma based on diagnostic and treatment norms in China.

Objectives: In this study, we compared the cost-effectiveness comparison of the active surveillance (AS) and early surgery (ES) approaches for papillary thyroid microcarcinoma (PTMC) from the perspective of the Chinese healthcare system. Methods: We performed a cost-effectiveness analysis using a Markov model of PTMC we developed to evaluate the incremental cost-effectiveness ratio of AS and ES. Our reference case was of a 40-year-old woman diagnosed with unifocal (<10 mm) PTMC. Relevant data were extracted after an extensive literature review, and the cost incurred in each state was determined using China Medicare data on payments for ES and AS. The willingness-to-pay threshold was set at ¥242,928/quality-adjusted life-year (QALY) gained. Sensitivity analyses were performed to account for any uncertainty in the model's variables. Additional subgroup analyses were performed to determine whether AS was cost-effective when different initial monitoring ages were used. Results: ES exhibited an effectiveness of 5.2 QALYs, whereas AS showed an effectiveness of 25.8 QALYs. Furthermore, the incremental cost-effectiveness ratio for ES versus AS was ¥1,009/QALY. The findings of all sensitivity analyses were robust. Compared with ES, AS was found to be the cost-effective strategy at initial monitoring ages of 20 and 60 years, with an incremental cost-effectiveness ratio of ¥3,431/QALY and -¥1,316/QALY at 20 and 60 years, respectively. AS was a more cost-effective strategy in patients with PTMC aged more than 60. Conclusions: With respect to the norms of the Chinese healthcare system, AS was more cost-effective for PTMC over lifetime surveillance than ES. Furthermore, it was cost-effective even when the initial monitoring ages were different. In addition, if AS is incorporated into the management plan for PTMC in China at the earliest possible stage, a predicted savings of ¥10 × 108/year could be enabled for every 50,000 cases of PTMC, which indicates a good economic return for future management programs. The identification of such nuances can help physicians and patients determine the best and most individualized long-term management strategy for low-risk PTMC.

Combined conventional ultrasonography with clinicopathological features to predict axillary status after neoadjuvant therapy for breast cancer: A case-control study.

OBJECTIVES: This study aimed to evaluate the value of a model combining conventional ultrasonography and clinicopathologic features for predicting axillary status after neoadjuvant therapy in breast cancer. METHODS: This retrospective study included 329 patients with lymph node-positive who underwent neoadjuvant systemic treatment (NST) from June 2019 to March 2022. Ultrasound and clinicopathological characteristics of breast lesions and axillary lymph nodes were analyzed before and after NST. The diagnostic efficacy of ultrasound, clinicopathological characteristics, and combined model were evaluated using multivariate logistic regression and receiver operator characteristic curve (ROC) analyses. RESULTS: The area under ROC (AUC) for the ability of the combined model to predict the axillary pathological complete response (pCR) after NST was 0.882, that diagnostic effectiveness was significantly better than that of the clinicopathological model (AUC of 0.807) and the ultrasound feature model (AUC of 0.795). In addition, eight features were screened as independent predictors of axillary pCR, including clinical N stage, ERBB2 status, Ki-67, and after NST the maximum diameter reduction rate and margins of breast lesions, the short diameter, cortical thickness, and fatty hilum of lymph nodes. CONCLUSIONS: The combined model constructed from ultrasound and clinicopathological features for predicting axillary pCR has favorable diagnostic results, which allowed more accurate identification of BC patients who had received axillary pCR after NST. ADVANCES IN KNOWLEDGE: A combined model incorporated ultrasound and clinicopathological characteristics of breast lesions and axillary lymph nodes demonstrated favorable performance in evaluating axillary pCR preoperatively and non-invasively.

Ultrasound and clinicopathological characteristics-based model for prediction of pathologic response to neoadjuvant chemotherapy in HER2-positive breast cancer: a case-control study.

BACKGROUND: The objective of this study was to develop a model combining ultrasound (US) and clinicopathological characteristics to predict the pathologic response to neoadjuvant chemotherapy (NACT) in human epidermal growth factor receptor 2 (HER2)-positive breast cancer. MATERIALS AND METHODS: This is a retrospective study that included 248 patients with HER2-positive breast cancer who underwent NACT from March 2018 to March 2022. US and clinicopathological characteristics were collected from all patients in this study, and characteristics obtained using univariate analysis at p < 0.1 were subjected to multivariate analysis and then the conventional US and clinicopathological characteristics independently associated with pathologic complete response (pCR) from the analysis were used to develop US models, clinicopathological models, and their combined models by the area under the receiver operating characteristic (ROC) curve (AUC), accuracy, sensitivity, and specificity to assess their predictive efficacy. RESULTS: The combined model had an AUC of 0.808, a sensitivity of 88.72%, a specificity of 60.87%, and an accuracy of 75.81% in predicting pCR of HER2-positive breast cancer after NACT, which was significantly better than the clinicopathological model (AUC = 0.656) and the US model (AUC = 0.769). In addition, six characteristics were screened as independent predictors, namely the Clinical T stage, Clinical N stage, PR status, posterior acoustic, margin, and calcification. CONCLUSION: The conventional US combined with clinicopathological characteristics to construct a combined model has a good diagnostic effect in predicting pCR in HER2-positive breast cancer and is expected to be a useful tool to assist clinicians in effectively determining the efficacy of NACT in HER2-positive breast cancer patients.

Value of Artificial Intelligence in Improving the Accuracy of Diagnosing TI-RADS Category 4 Nodules.

OBJECTIVE: Considerable heterogeneity is observed in the malignancy rates of thyroid nodules classified as category 4 according to the Thyroid Imaging Reporting and Data System (TI-RADS). This study was aimed at comparing the diagnostic performance of artificial intelligence algorithms and radiologists with different experience levels in distinguishing benign and malignant TI-RADS 4 (TR4) nodules. METHODS: Between January 2019 and September 2022, 1117 TR4 nodules with well-defined pathological findings were collected for this retrospective study. An independent external data set of 125 TR4 nodules was incorporated for testing purposes. Traditional feature-based machine learning (ML) models, deep convolutional neural networks (DCNN) models and a fusion model that integrated the prediction outcomes from all models were used to classify benign and malignant TR4 nodules. A fivefold cross-validation approach was employed, and the diagnostic performance of each model and radiologists was compared. RESULTS: In the external test data set, the area under the receiver operating characteristic curve (AUROC) of the three DCNN-based secondary transfer learning models-InceptionV3, DenseNet121 and ResNet50-were 0.852, 0.837 and 0.856, respectively. These values were higher than those of the three traditional ML models-logistic regression, multilayer perceptron and random forest-at 0.782, 0.790, and 0.767, respectively, and higher than that of an experienced radiologist (0.815). The fusion diagnostic model we developed, with an AUROC of 0.880, was found to outperform the experienced radiologist in diagnosing TR4 nodules. CONCLUSION: The integration of artificial intelligence algorithms into medical imaging studies could improve the accuracy of identifying high-risk TR4 nodules pre-operatively and have significant clinical application potential.

Relationship between inflammatory-related cytokines with aortic dissection.

Aortic dissection, characterized by severe intramural hematoma formation and acute endometrial rupture, is caused by excessive bleeding within the aortic wall or a severe tear within the intimal layer of the aorta, which subsequently promotes the separation or dissection in the layers of the aortic wall. Epidemiological surveys showed that aortic dissection was most observed among those patients from 55 to 80 years of age, with a prevalence of approximately 40 cases per 100,000 individuals per year, posing serious risks to future health and leading to high mortality. Other risk factors of aortic dissection progression contained dyslipidemia, hypertension, and genetic disorders, such as Marfan syndrome. Currently, emerging evidence indicates the pathological progression of aortic dissection is significantly complicated, which is correlated with the aberrant infiltration of pro-inflammatory cells into the aortic wall, subsequently facilitating the apoptosis of vascular smooth muscle cells (VSMCs) and inducing the aberrant expression of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and interferon (IF). Other pro-inflammatory-related cytokines, including the colony-stimulating factor (CSF), chemotactic factor, and growth factor (GF), played an essential function in facilitating aortic dissection. Multiple studies focused on the important relationship between pro-inflammatory cytokines and aortic dissection, which could deepen the understanding of aortic dissection and further guide the therapeutic strategies in clinical practice. The present review elucidated pro-inflammatory cytokines' functions in modulating the risk of aortic dissection are summarized. Moreover, the emerging evidence that aimed to elucidate the potential mechanisms wherebyvarious pro-inflammatory cytokines affected the pathological development of aortic dissection was also listed.

Porcine Reproductive and Respiratory Syndrome Virus Modulates the Switch of Macrophage Polarization from M1 to M2 by Upregulating MoDC-Released sCD83.

Porcine reproductive and respiratory syndrome virus (PRRSV), the most economically important infectious disease of pigs, elicits poor innate and adaptive immune responses. Soluble CD83 (sCD83), a secretion from various immune cell populations, especially MoDCs, is involved in negatively regulating the immune response. We speculate sCD83 may be a critical factor in the process of PRRSV-coordinated macrophage polarization. In this study, we found that PAMs co-cultured with PRRSV-infected MoDCs inhibited the M1 macrophage while enhancing the M2 macrophage. This was accompanied by a decrease in the pro-inflammatory cytokine TNF-α and iNOS and an increase in the anti-inflammatory cytokine IL-10 and Arg1. Meanwhile, sCD83 incubation causes the same specific effects lead to a switch in macrophage from M1 to M2. Neutralization of sCD83 removes the inhibitory effects of PRRSV on PAMs. Using reverse genetics, we generated recombinant PRRSVs with mutations in N protein, nsp1α, and nsp10 (knockout sCD83-concerned key amino acid site). Four mutant viruses lost the suppression of M1 macrophage markers, in contrast to the restriction of the upregulation of M2 macrophage markers. These findings suggest that PRRSV modulates the switch of macrophage polarization from M1 to M2 by upregulating the MoDC-induced secretion of CD83, providing new insights into the mechanism by which PRRSV regulates host immunity.

Apolipoprotein A1 Inhibits Adipogenesis Progression of Human Adipose-Derived Mesenchymal Stem Cells.

BACKGROUND: According to the reports, the most vital characteristic of obesity is an aberrant accumulation of triglycerides (TG) in the adipocyte. On the other hand, circulating concentrations of apolipoprotein A1 (apoA1) have been demonstrated to be strongly correlated with the prevalence and the pathological development of obesity. Nevertheless, the underlying mechanisms whereby apoA1 modulates the pathogenesis of obesity is still not fully elucidated. METHODS: Adipose-derived mesenchymal stem cells (AMSCs, isolated from the hospitalized patients were combined with 15 µg/ml recombined human apoA1 protein. The effects of apoA1 on modulating the intracellular levels of TG and the expression contents of adipogenic related cytokines were also analyzed. Furthermore, whether apoA1 modulated the adipogenesis progression via sortilin was also explored in the current research. RESULTS: During the adipogenesis progression, apoA1 could significantly lower the quantity of intracellular lipid droplets (LDs). Meanwhile, apoA1 could decrease the intracellular levels of TG and down-regulate the expression contents of several vital adipogenic related cytokines, such as CCAAT enhancer-binding proteins α/ß (C/EBPα/ß), fatty acid synthetase (FAS), and fatty acid-binding protein 4 (FABP4). Moreover, the inhibitory effect of apoA1 was further verified to be induced through upregulating the SORT1 gene expression which subsequently increased sortilin protein. Consistent with these findings, silencing the SORT1 gene expression could induce the loss-of-function (LOF) of apoA1 in modulating the adipogenesis progression of AMSCs. CONCLUSION: In conclusion, apoA1 could suppress the adipogenesis progression of human AMSCs through, at least partly, up-regulating the SORT1 gene expression which subsequently increases the sortilin protein content. Thereby, the present research sheds light on a novel pathogenic mechanism by which apoA1 regulates adipogenesis progression and proposes that apoA1 embraces the function to treat obesity in clinical practice.

Peptide GL13K releasing hydrogel functionalized micro/nanostructured titanium enhances its osteogenic and antibacterial activity.

Bacterial infection and the subsequent lack of osseointegration are two of the main problems encountered during the early stage of implantation. To resolve these issues, in this study, the micro/nanostructure on titanium surface was fabricated by dual-acid treatment and anodic oxidation, and then GelMA hydrogel loaded with GL13K was coated onto the micro/nanostructured titanium. The surface properties of different titanium substrates were characterized by scanning electron microscopy, atomic force microscope, water contact angle measurement, and in vitro release of GL13K. The results demonstrated that peptide GL13K releasing hydrogel was successfully modified onto the micro/nanostructured titanium and GL13K displayed an effective controlled release. In vitro cell experiments including cytoskeleton observation, cell viability, phosphatase activity, mineralization and the expression of osteogenic related genes proved that the peptide GL13K releasing hydrogel modified micro/nanostructured titanium had good biocompatibility, promoted osteoblast differentiation. In vitro antibacterial experiments showed that it also can prevent the growth of gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria by the release of GL13K. This study demonstrates titanium surface modification that allows for the development of anti-bacterial titanium implants.

MiR-424-5p targets HSP90AA1 to facilitate proliferation and restrain differentiation in skeletal muscle development.

MiR-424-5p was found to be a potential regulator in the proliferation, migration, and invasion of various cancer cells. However, the effects and functional mechanism of miR-424-5p in the process of myogenesis are still unclear. Previously, using microRNA (miRNA) sequencing and expression analysis, we discovered that miR-424-5p was expressed differentially in the different skeletal muscle growth periods of Xuhuai goats. We hypothesized that miR-424-5p might play an important role in skeletal muscle myogenesis. Then, we found that the proliferation ability of the mouse myoblast cell (C2C12 myoblast cell line) was significantly augmented, whereas the C2C12 differentiation was repressed after increasing the expression of miR-424-5p. Mechanistically, HSP90AA1 presented a close interrelation with miR-424-5p, which was predicted as a target gene in the progression of skeletal muscle myogenesis, using transcriptome sequencing, dual luciferase reporter gene detection, and qRT-PCR. The silencing of HSP90AA1 obviously increased C2C12 proliferation and diminished differentiation, which is consistent with the ability of miR-424-5p in C2C12. Altogether, our findings indicated the role of miR-424-5p as a novel potential regulator via HSP90AA1 during muscle myogenesis progression.

Association of MBOAT7 rs641738 polymorphism with hepatocellular carcinoma susceptibility: A systematic review and meta-analysis.

BACKGROUND: The MBOAT7 rs641738 single-nucleotide polymorphism (SNP) has been proven to influence various liver diseases, but its association with hepatocellular carcinoma (HCC) susceptibility has been debated. To address this discrepancy, we conducted the current systematic review and meta-analysis. AIM: To perform a systematic review and meta-analysis on association of MBOAT7 SNP and HCC susceptibility. METHODS: We performed a systematic review in PubMed, Web of Science, Scopus, and EMBASE; applied specific inclusion and exclusion criteria; and extracted the data. Meta-analysis was conducted with the meta package in R. Sensitivity and subgroup analyses were also performed. This meta-analysis was registered in PROSPERO (CRD42023458046). RESULTS: Eight studies were included in the systematic review, and 12 cohorts from 6 studies were included in the meta-analysis. Our meta-analysis revealed an association between the MBOAT7 SNP and HCC susceptibility in both the dominant [odds ratio (OR): 1.14, 95% confidence interval (95%CI): 1.02-1.26, P = 0.020] and recessive (OR: 1.21, 95%CI: 1.05-1.39, P = 0.008) models. Subgroup analysis revealed that stratification of the included patients by geographical origin showed a significant association in Asia (OR: 1.20, 95%CI: 1.03-1.39). CONCLUSION: This meta-analysis underscores the contribution of the MBOAT7 rs641738 SNP to hepatocarcinogenesis, especially in Asian populations, which warrants further investigation.

Clinical analysis of 13 cases of primary squamous-cell thyroid carcinoma.

Objective: To analyze the clinical features, ultrasonographic manifestations, pathological features, treatment and prognosis of primary thyroid squamous cell carcinoma (PSCTC) and summarize the experience in diagnosis and treatment of this condition. Methods: A retrospective analysis was conducted on patients who were admitted to Zhejiang Cancer Hospital from 2007 to 2021 due to thyroid nodules or thyroid malignant tumors that were ultimately confirmed by postoperative pathology as primary thyroid squamous cell carcinoma. We summarize the general situation, clinical information, laboratory examination, ultrasonic image characteristics, pathological examination, clinical treatment and prognosis of the patients. Results: PSCTC is most often seen in older men and progresses rapidly. In laboratory tests, some patients had elevated levels of tumor markers (CA199, squamous cell carcinoma antigen level), thyroglobulin levels and tumor-related substances, but all these indicators lacked specificity. The ultrasound features of PSCTC are mainly hypoechoic, hard, substantial nodules with gross borders and a grade 1-2 blood flow signal, sometimes with signs of necrosis and calcification. In terms of treatment, PSCTC is mainly surgically resected, though some patients in this study underwent iodine-131 radiation therapy, local radiotherapy, and chemotherapy with unclear results. None of the patients survived for very long after treatment, but the prognosis of patients with highly differentiated squamous carcinoma was significantly better than that of patients with poorly differentiated squamous carcinoma. Papillary thyroid carcinoma may be one of the causes of PSCTC. Conclusion: PSCTC is a malignant tumor with high malignancy and rapid clinical progression. Treatment options are mainly based on surgical resection and can be supplemented with radiotherapy and chemotherapy, but there is still a lack of a standardized treatment management system, and more cases and reports are needed to accumulate data.

Improved Soluble Expression in Escherichia coli and Easily Purified Recombinant Human Bone Morphogenetic 7-2 Fusion Protein.

BACKGROUND: Bone morphogenetic protein (BMP) is a cysteine-rich growth factor and plays a key role in early bone tissue development and bone defect repair. However, the low yield, high cost and complicated process in BMP significantly limit its clinical application. OBJECTIVE: In this study, we developed an efficient method for soluble expression and preparation of recombinant human bone morphogenetic 7-2 fusion protein (rhBMP7-2) and determined its molecular weight and biological activity. METHODS: The fusion gene for rhBMP-2 and rhBMP-7 was inserted into the pET-ELP expression vector. Correct DNA sequence was confirmed, the rhBMP7-2-ELP was transformed into Escherichia coli strain BL21 (DE3), and the rhBMP7-2 was produced in the recombinant E. coli. Recombinant BMP7-2 purify was identified using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). The cell proliferation and biological activity of rhBMP7-2 were measured by Cell Counting Kit-8 and Alkaline Phosphatase assay using C2C12 cells, respectively. RESULTS: The result of digestion of NdeI, BamHI and XhoI enzymes showed that the rhBMP7-2-ELP was correctly constructed. The recombinant BMP7-2 was successfully expressed in soluble form; the purified rhBMP7-2 showed biological activity and significantly promoted cell proliferation and differentiation in a dose-dependent manner. CONCLUSION: The rhBMP7-2 fusion protein with osteogenic activity was prepared through a lowcost and time-efficient method. Our preparation method presents the potential to be applied to the large-scale production of rhBMP7-2 and is expected to play a significant role in clinical treatment.

Integration of Interleukin-6 Improves the Diagnostic Precision of Metagenomic Next-Generation Sequencing for Infection in Immunocompromised Children.

The performance of metagenomic next-generation sequencing (mNGS) in identifying pathogens in immunocompromised children was not very clear. The purpose of this study is to assess the performance of mNGS in this population and to investigate whether the integration of serum cytokines and mNGS assay could improve diagnostic accuracy. We retrospectively collected the clinical data of pediatric patients who suffered febrile diseases and underwent mNGS determination simultaneously in the department of hematology/oncology between January 2019 and March 2021. Specimens were sent for conventional microbiological test (CMT), mNGS, and serum cytokine measurement in parallel. A total of 258 episodes of febrile diseases were enrolled, mNGS was positive in 224 cases, while CMT was positive in 78 cases. mNGS and CMT were both positive in 70 (27.1%) cases and were both negative in 26 (10.1%) cases. There were 154 (59.7%) cases positive by mNGS only while 8 (3.1%) were positive by CMT only. It was common that two or more pathogens were simultaneously detected by mNGS in a single specimen, with only 61 tests identified a single organism. Whether the organisms reported by mNGS were the microbiological etiology of infection was evaluated. Of the 224 cases with positive mNGS results, 135 (58.4%), 30 (13.0%), and 59 (28.6%) were considered as "probable," "possible," and "unlikely," respectively. Patients with high IL-6 (≥ 390 pg/ml) were likely to be bacterial infection. Although mNGS reported mixed pathogens, 84.6% (33/39) and 83.3% (10/12) of patients presenting high IL-6 were confirmed as bacterial infection in the training and validation cohort, respectively. In conclusion, mNGS analysis demonstrates promising diagnostic potential in rapidly identifying clinically relevant pathogens. Given the detection of many clinically irrelevant organisms, the integration of IL-6 improves the precision of mNGS results interpretation.

Ex Vivo Expansion and CRISPR-Cas9 Genome Editing of Primary Human Natural Killer Cells.

Natural killer (NK) cells are potent innate immune cells that provide the surveillance and elimination of infected, stressed, and malignant cells. The unique immune recognition mechanisms and functions of NK cells make them an attractive cell type for immunology research and adoptive immunotherapy. However, primary NK cells are challenging to culture ex vivo and lack efficient genetic tools, hindering the research of NK cells and the development of NK cell therapeutics. Here we describe methods for the freeze-thaw process, feeder-free ex vivo expansion, CRISPR-Cas9 genome editing, and functional characterizations of primary human NK cells. Our protocol enables ∼30-fold and ∼2000-fold average expansion rates from 1 × 107 cryopreserved NK cells in 14 and 28 days, respectively. We also detail methods for CRISPR gene knockout and knockin by nucleofection of Cas9 ribonucleoproteins (RNP) and DNA repair templates. Gene knockout by Cas9 RNP nucleofection can be multiplexed to simultaneously target three genes. The CRISPR-edited cells can be cryopreserved and rethawed with high viability for future studies. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Thawing of natural killer cells Basic Protocol 2: Ex vivo expansion of natural killer cells Basic Protocol 3: Cryopreservation of expanded natural killer cells Basic Protocol 4: Characterization of natural killer cells: Flow cytometry and surface marker analysis Basic Protocol 5: Cytotoxicity and degranulation assays Basic Protocol 6: Preparation of homology-directed repair templates Basic Protocol 7: Nucleofection of CRISPR-Cas9 ribonucleoproteins Basic Protocol 8: Genotyping of gene-edited natural killer cells Basic Protocol 9: Phenotyping of gene-edited natural killer cells.

The Effect of 5A nursing intervention on living quality and self-care efficacy of patients undergoing chemotherapy after hepatocellular carcinoma surgery.

OBJECTIVES: This paper explored and analyzed the effect of 5A nursing intervention on living quality and self-care efficacy of patients undergoing postoperative chemotherapy after hepatocellular carcinoma surgery. METHODS: A total of 97 patients with hepatocellular carcinoma hospitalized from March 2017 to June 2019 were chosen as subjects and separated to the control group (n=47) and observation group (n=50) based on the admission time. The control-group received a conventional nursing care mode, and the observation-group was given the 5A nursing care mode in addition to the conventional care mode in the control group. Self-care efficacy, SF-36 quality of life, cancer-related fatigue degree and satisfaction with nursing work of patients pre- and post-intervention were compared between the two groups. RESULTS: The self-care efficacy of the two groups post-intervention was critically higher than during pre-intervention (P<0.05), and the index of the observation group was significantly higher than that of the control group (P<0.05). The scores in each dimension of life quality in both groups after intervention were remarkably higher than those for pre-intervention (P<0.05), and the index in observation group was notably higher than that in the control group (P<0.05). Cancer-related fatigue degree of the two groups of patients during post-intervention was critically higher than those in pre-intervention (P<0.05), while during post-intervention, the index in control group was notably higher than in observation group (P<0.05). The satisfaction degree of nursing skills, service attitude, health training and hospital condition in the observation group were significantly higher than those in the control group (P<0.05). CONCLUSION: The model of 5A nursing intervention can ameliorate the self-care efficacy and living quality of patients undergoing chemotherapy after hepatocellular carcinoma surgery. The intervention can also help alleviate cancer-related fatigue during treatment, and improve the satisfaction degree of patients with nursing care, which is worthy of clinical promotion and application.

Optimal timing for hospice-shared care initiation in terminal cancer patients.

PURPOSE: The existing concept suggests early palliative and hospice therapy for a better quality of care (QOC) and less medical expense in terminal cancer patients, but the time points of "early" initiation were defined by pre-set study protocol rather than the real-world data. The study aimed to determine the optimal timing of initiating palliative care for patients with terminal cancer. METHODS: This retrospective population-based study was conducted using a nationwide database. We extracted patients with cancer who were in their last year of lives in the period from 1 January 2010 to 31 December 2013 and categorized them into two groups ("hospice-shared care" (HSC) group and "usual care" (UC) group) after a matching process. Subsequently, we used a generalized linear mixed-effects model to compare the QOC and medical expenses between groups. RESULTS: After the selection and matching process, we enrolled 1714 patients (67.7 ± 13.2 years, 62.7% male) categorized into the HSC and UC groups (n = 857 in each group). The HSC groups showed generally better QOC in the four indices (with emergency room visit, hospitalization, intensive care unit admission, and receiving chemotherapy) than the UC group in those who initiated HSC 8-60 days before death. The HSC group also had significantly lower medical expenses than the UC group in those who initiated HSC 15-90 days before death. CONCLUSIONS: Among patients with terminal cancer, HSC initiation before the last 8 days and 15 days of lives can effectively improve QOC and save medical expenses, respectively.

Surface Expression of Kynurenine 3-Monooxygenase Promotes Proliferation and Metastasis in Triple-Negative Breast Cancers.

Kynurenine 3-monooxygenase (KMO) is the pivotal enzyme in the kynurenine pathway and is located on the mitochondrial outer membrane. The dysregulation of KMO leads to various neurodegenerative diseases; however, it is rarely mentioned in cancer progression. Our previous study showed that KMO overexpression in canine mammary gland tumors (cMGT) is associated with poor prognosis in cMGT patients. Surprisingly, it was also found that KMO can be located on the cell membranes of cMGT cells, unlike its location in normal cells, where KMO is expressed only within the cytosol. Since cMGT and human breast cancer share similar morphologies and pathogenesis, this study investigated the possibility of detecting surface KMO in human breast cancers and the role of surface KMO in tumorigenesis. Using immunohistochemistry (IHC), flow cytometry (FC), immunofluorescence assay (IFA), and transmission electron microscopy (TEM), we demonstrated that KMO can be aberrantly and highly expressed on the cell membranes of breast cancer tissues and in an array of cell lines. Masking surface KMO with anti-KMO antibody reduced the cell viability and inhibited the migration and invasion of the triple-negative breast cancer cell line, MDA-MB-231. These results indicated that aberrant surface expression of KMO may be a potential therapeutic target for human breast cancers.