Manufacturing, quality control, and GLP-grade preclinical study of nebulized allogenic adipose mesenchymal stromal cells-derived extracellular vesicles.

BACKGROUND: Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, quality control (QC) and preclinical safety of clinical-grade haMSC-EVs. METHODS: haMSC-EVs were isolated from the conditioned medium of human adipose MSCs incubated in 2D containers. Purification was performed by PEG precipitation and differential centrifugation. Characterizations were conducted by nanoparticle tracking analysis, transmission electron microscopy (TEM), Western blotting, nanoflow cytometry analysis, and the TNF-α inhibition ratio of macrophage [after stimulated by lipopolysaccharide (LPS)]. RNA-seq and proteomic analysis with liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to inspect the lot-to-lot consistency of the EV products. Repeated toxicity was evaluated in rats after administration using trace liquid endotracheal nebulizers for 28 days, and respiratory toxicity was evaluated 24 h after the first administration. In vivo therapeutic effects were assessed in an LPS-induced ALI/ acute respiratory distress syndrome (ARDS) rat model. RESULTS: The quality criteria have been standardized. In a stability study, haMSC-EVs were found to remain stable after 6 months of storage at - 80°C, 3 months at - 20 °C, and 6 h at room temperature. The microRNA profile and proteome of haMSC-EVs demonstrated suitable lot-to-lot consistency, further suggesting the stability of the production processes. Intratracheally administered 1.5 × 108 particles/rat/day for four weeks elicited no significant toxicity in rats. In LPS-induced ALI/ARDS model rats, intratracheally administered haMSC-EVs alleviated lung injury, possibly by reducing the serum level of inflammatory factors. CONCLUSION: haMSC-EVs, as an off-shelf drug, have suitable stability and lot-to-lot consistency. Intratracheally administered haMSC-EVs demonstrated excellent safety at the tested dosages in systematic preclinical toxicity studies. Intratracheally administered haMSC-EVs improved the lung function and exerted anti-inflammatory effects on LPS-induced ALI/ARDS model rats.

Cytosolic Cadherin 4 promotes angiogenesis and metastasis in papillary thyroid cancer by suppressing the ubiquitination/degradation of ß-catenin.

BACKGROUND: Although the long-term prognosis of papillary thyroid cancer (PTC) is favorable, distant metastasis significantly compromises the prognosis and quality of life for patients with PTC. The Cadherin family plays a pivotal role in tumor metastasis; however, the involvement of Cadherin 4 (CDH4) in the metastatic cascade remains elusive. METHODS: The expression and subcellular localization of CDH4 were determined through immunohistochemistry, immunofluorescence, and western blot analyses. The impact of CDH4 on cell migration, invasion, angiogenesis, and metastasis was assessed using transwell assays, tube formation assays, and animal experiments. Immunoprecipitation assay and mass spectrometry were employed to examine protein associations. The influence of CDH4 on the subcellular expression of ß-catenin and active ß-catenin was investigated via western blotting and immunofluorescence. Protein stability and ubiquitination assay were employed to verify the impact of CDH4 on ß-catenin degradation. Rescue experiments were performed to ensure the significance of CDH4 in regulating nuclear ß-catenin signaling. RESULTS: CDH4 was found to be significantly overexpressed in PTC tissues and predominantly localized in the cytoplasm. Furthermore, the overexpression of CDH4 in tumor tissues is associated with lymph node metastasis in PTC patients. Cytosolic CDH4 promoted the migration, invasion, and lung metastasis of PTC cells and stimulated the angiogenesis and tumorigenesis of PTC; however, this effect could be reversed by Tegavivint, an antagonist of ß-catenin. Mechanistically, cytosolic CDH4 disrupted the interaction between ß-catenin and ß-TrCP1, consequently impeding the ubiquitination process of ß-catenin and activating the nuclear ß-catenin signaling. CONCLUSIONS: CDH4 induces PTC angiogenesis and metastasis via the inhibition of ß-TrCP1-dependent ubiquitination of ß-Catenin.

Added value of arterial enhancement fraction derived from dual-energy computed tomography for preoperative diagnosis of cervical lymph node metastasis in papillary thyroid cancer: initial results.

OBJECTIVES: To explore the added value of arterial enhancement fraction (AEF) derived from dual-energy computed tomography CT (DECT) to conventional image features for diagnosing cervical lymph node (LN) metastasis in papillary thyroid cancer (PTC). METHODS: A total of 273 cervical LNs (153 non-metastatic and 120 metastatic) were recruited from 92 patients with PTC. Qualitative image features of LNs were assessed. Both single-energy CT (SECT)-derived AEF (AEFS) and DECT-derived AEF (AEFD) were calculated. Correlation between AEFD and AEFS was determined using Pearson's correlation coefficient. Multivariate logistic regression analysis with the forward variable selection method was used to build three models (conventional features, conventional features + AEFS, and conventional features + AEFD). Diagnostic performances were evaluated using receiver operating characteristic (ROC) curve analyses. RESULTS: Abnormal enhancement, calcification, and cystic change were chosen to build model 1 and the model provided moderate diagnostic performance with an area under the ROC curve (AUC) of 0.675. Metastatic LNs demonstrated both significantly higher AEFD (1.14 vs 0.48; p < 0.001) and AEFS (1.08 vs 0.38; p < 0.001) than non-metastatic LNs. AEFD correlated well with AEFS (r = 0.802; p < 0.001), and exhibited comparable performance with AEFS (AUC, 0.867 vs 0.852; p = 0.628). Combining CT image features with AEFS (model 2) and AEFD (model 3) could significantly improve diagnostic performances (AUC, 0.865 vs 0.675; AUC, 0.883 vs 0.675; both p < 0.001). CONCLUSIONS: AEFD correlated well with AEFS, and exhibited comparable performance with AEFS. Integrating qualitative CT image features with both AEFS and AEFD could further improve the ability in diagnosing cervical LN metastasis in PTC. CLINICAL RELEVANCE STATEMENT: Arterial enhancement fraction (AEF) values, especially AEF derived from dual-energy computed tomography, can help to diagnose cervical lymph node metastasis in patients with papillary thyroid cancer, and complement conventional CT image features for improved clinical decision making. KEY POINTS: • Metastatic cervical lymph nodes (LNs) demonstrated significantly higher arterial enhancement fraction (AEF) derived from dual-energy computed tomography (DECT) and single-energy CT (SECT)-derived AEF (AEFS) than non-metastatic LNs in patients with papillary thyroid cancer. • DECT-derived AEF (AEFD) correlated significantly with AEFS, and exhibited comparable performance with AEFS. • Integrating qualitative CT images features with both AEFS and AEFD could further improve the differential ability.

Influence of sex, age and thyroid function indices on dual-energy computed tomography-derived quantitative parameters of thyroid in patients with or without Hashimoto's thyroiditis.

PURPOSE: To study the influence of sex, age and thyroid function indices on dual-energy computed tomography (DECT)-derived quantitative parameters of thyroid in patients with or without Hashimoto's thyroiditis (HT). MATERIAL AND METHODS: A total of 198 consecutive patients who underwent DECT scan of neck due to unilateral thyroid lesions were retrospectively enrolled. Iodine concentration (IC), total iodine content (TIC) and volume of normal thyroid lobe were calculated. Influences of sex, age and thyroid function indices on DECT-derived parameters in overall study population, subgroup patients with, and those without HT were assessed using Mann-Whitney U test, Student's T-test, and Spearman correlation analyses, respectively, as appropriate. RESULTS: HT group showed significantly lower IC and TIC, while higher volume than No-HT group (all p < 0.001). The volume was larger in male than that in female in overall study population and No-HT group (p = 0.047 and 0.010, respectively). There was no significant difference in any DECT-derived parameters between low (≤ 35 years) and high (> 35 years) age group in all three groups (all p > 0.05). TPOAb and TgAb correlated positively with IC and TIC, and negatively with volume in overall study population (all p < 0.05). TPOAb and TgAb also correlated positively with IC in HT group (p = 0.002 and 0.007, respectively). CONCLUSION: DECT-derived parameters of thyroid differed significantly between patients with and without HT. Sex and thyroid function indices could affect the DECT-derived parameters. Aforementioned physiological factors should be considered when analyzing the DECT-derived parameters of thyroid.

Cell diversity and immune infiltration in the parathyroid tumour microenvironment.

Tumour microenvironment has been recognized as a crucial factor influencing disease progression. However, relevant features and functions are insufficiently understood in parathyroid neoplasia. Single-cell RNA sequencing was performed to profile the transcriptome of 27,251 cells from 4 parathyroid adenoma (PA) tissue samples. External transcriptomic datasets and immunofluorescence staining of a tissue microarray were set for expression validation. Eight major cell types and various subpopulations were finely identified in PA. We found that a subcluster of tumour endocrine cells with low copy number variation probably presented as a resting state. Diverse infiltrating immune cell subtypes were identified, constructing an immunosuppressive microenvironment. Tumour-associated macrophages, which indicated an anti-inflammatory phenotype, were significantly increased in PA. Inflammatory tumour-associated fibroblasts (iTAFs) were newly verified and highlighted on the role of stromal-immune crosstalk. Positive correlation between iTAFs and increased CD163+ macrophages was uncovered. Moreover, CXCL12 receptor signalling is important for tumour angiogenesis and immune infiltration. Our findings provide a comprehensive landscape interpreting tumour cell heterogeneity, cell diversity, and immune regulation in parathyroid neoplasia. The valuable resources may promote the understanding of parathyroid tumour microenvironment.

Right time to detect urine iodine during papillary thyroid carcinoma diagnosis and treatment: A case report.

BACKGROUND: This is the first documentation of a spontaneous and nonspecific chemical reaction of an iodinated contrast media with ammonium persulfate used in As3+-Ce4+ catalytic spectrophotometry for urine iodine concentration (UIC) detection. CASE SUMMARY: We herein report an incidental case who had a dual source computed tomography examination for papillary thyroid carcinoma diagnosis. Serial spot urine specimens were collected during her hospitalization and were measured by As3+-Ce4+ catalytic spectrophotometry on a Beckman Coulter AU5800. The reacted solutions were "brownish", and the results showed extremely high iodine concentrations despite serial dilutions. The patient claimed no dietary habit of iodized salt or iodine-containing medical history, which strongly pointed to iodinated contrast media (ICM) via intravenous injection. Even with 0.01% ICM, its interruption is still profound on the desired urine iodine reaction with ammonium persulfate, leading to inaccurate UIC and possibly inappropriate treatment. CONCLUSION: The following laboratory suggestions should be considered: (1) As3+-Ce4+ catalytic spectrophotometry is only suitable for UIC measurement after confirmed ICM renal clearance; (2) A mass spectrometry-based method can be applied as an alternative during the ICM clearance period; and (3) The UIC baseline can be confirmed after ICM injection by consecutive detection for at least 2 mo.

Extracellular Volume Fraction Derived From Dual-Layer Spectral Detector Computed Tomography for Diagnosing Cervical Lymph Nodes Metastasis in Patients With Papillary Thyroid Cancer: A Preliminary Study.

Objectives: The current study evaluates the performance of dual-energy computed tomography (DECT) derived extracellular volume (ECV) fraction based on dual-layer spectral detector CT for diagnosing cervical lymph nodes (LNs) metastasis from papillary thyroid cancer (PTC) and compares it with the value of ECV derived from conventional single-energy CT (SECT). Methods: One hundred and fifty-seven cervical LNs (81 non-metastatic and 76 metastatic) were recruited. Among them, 59 cervical LNs (27 non-metastatic and 32 metastatic) were affected by cervical root artifact on the contrast-enhanced CT images in the arterial phase. Both the SECT-derived ECV fraction (ECVS) and the DECT-derived ECV fraction (ECVD) were calculated. A Pearson correlation coefficient and a Bland-Altman analysis were performed to evaluate the correlations between ECVD and ECVS. Receiver operator characteristic curves analysis and the Delong method were performed to assess and compare the diagnostic performance. Results: ECVD correlated significantly with ECVS (r = 0.925; p <0.001) with a small bias (-0.6). Metastatic LNs showed significantly higher ECVD (42.41% vs 22.53%, p <0.001) and ECVS (39.18% vs 25.45%, p <0.001) than non-metastatic LNs. By setting an ECVD of 36.45% as the cut-off value, optimal diagnostic performance could be achieved (AUC = 0.813), which was comparable with that of ECVS (cut-off value = 34.99%; AUC = 0.793) (p = 0.265). For LNs affected by cervical root artifact, ECVD also showed favorable efficiency (AUC = 0.756), which was also comparable with that of ECVS (AUC = 0.716) (p = 0.244). Conclusions: ECVD showed a significant correlation with ECVS. Compared with ECVS, ECVD showed comparable performance in diagnosing metastatic cervical LNs in PTC patients, even though the LNs were affected by cervical root artifacts on arterial phase CT.

Nebulized exosomes derived from allogenic adipose tissue mesenchymal stromal cells in patients with severe COVID-19: a pilot study.

BACKGROUND: Existing clinical studies supported the potential efficacy of mesenchymal stromal cells as well as derived exosomes in the treatment of COVID-19. We aimed to explore the safety and efficiency of aerosol inhalation of the exosomes derived from human adipose-derived MSCs (haMSC-Exos) in patients with COVID-19. METHODS: The MEXCOVID trial is a phase 2a single-arm, open-labelled, interventional trial and patients were enrolled in Jinyintan Hospital, Wuhan, China. Eligible 7 patients were assigned to receive the daily dose of haMSCs-Exos (2.0 × 108 nano vesicles) for consecutively 5 days. The primary outcomes included the incidence of prespecified inhalation-associated events and serious adverse events. We also observed the demographic data, clinical characteristics, laboratory results including lymphocyte count, levels of D-dimer and IL-6 as well as chest imaging. RESULTS: Seven severe COVID-19 related pneumonia patients (4 males and 3 females) were enrolled and received nebulized haMSC-Exos. The median age was 57 year (interquartile range (IQR), 43 year to 70 year). The median time from onset of symptoms to hospital admission and administration of nebulized haMSC-Exos was 30 days (IQR, 15 days to 40 days) and 54 d (IQR, 34 d to 69 d), respectively. All COVID-19 patients tolerated the haMSC-Exos nebulization well, with no evidence of prespecified adverse events or clinical instability during the nebulization or during the immediate post-nebulization period. All patients presented a slight increase of serum lymphocyte counts (median as 1.61 × 109/L vs. 1.78 × 109/L). Different degrees of resolution of pulmonary lesions after aerosol inhalation of haMSC-Exos were observed among all patients, more obviously in 4 of 7 patients. CONCLUSIONS: Our trial shows that a consecutive 5 days inhalation dose of clinical grade haMSC-Exos up to a total amount of 2.0 × 109 nano vesicles was feasible and well tolerated in seven COVID-19 patients, with no evidence of prespecified adverse events, immediate clinical instability, or dose-relevant toxicity at any of the doses tested. This safety profile is seemingly followed by CT imaging improvement within 7 days. Further trials will have to confirm the long-term safety or efficacy in larger population. TRIAL REGISTRATION: MEXCOVID, NCT04276987.

Proactive exploration of inferior parathyroid gland using a novel meticulous thyrothymic ligament dissection technique.

INTRODUCTION: The inferior parathyroid gland (IPTG) is widely distributed; effective techniques for its safe exploration and protection during thyroid surgery have not been documented. The thyrothymic ligament (TTL) is a connective tissue located between the thymic tongue and thyroid. This study aims to introduce a novel meticulous thyrothymic ligament dissection technique and assess its role in proactive exploration and situ preservation of IPTG. MATERIALS AND METHODS: 737 patients undergoing initial thyroid surgery between 2017 and 2021 in the Department of General Surgery of the First Affiliated Hospital of Nanjing Medical University were retrospectively recruited for this clinical study. In 391 of the recruited patients, the TTL was dissected, and the number and location of IPTG were recorded. Among them, 214 patients underwent total/near-total thyroidectomy (TT) plus central neck dissection (CND) were assigned to the observation group. The control group included 346 consecutive patients who underwent conventional TT plus CND. After 1:1 propensity score matching, each group contained 206 patients. The incidence of postoperative hypoparathyroidism was recorded. RESULTS: Among the 391 patients, 596 sides were dissected, out of which 436 sides (73.2%) had TTL, and approximately 90.1% of IPTG were located and identified. A statistically significant difference in incidence of temporary (27.7 vs. 49.0%, P < 0.001) and permanent hypoparathyroidism (0 vs. 8.2%, P = 0.047) was noted between the observation group and the control group. CONCLUSION: The meticulous thyrothymic ligament dissection technique helps to protect IPTG in situ and reduce the incidence of postoperative hypoparathyroidism.

Radiomics from Primary Tumor on Dual-Energy CT Derived Iodine Maps can Predict Cervical Lymph Node Metastasis in Papillary Thyroid Cancer.

RATIONALE AND OBJECTIVES: To develop and validate 2 iodine maps based radiomics nomograms for preoperatively predicting cervical lymph node metastasis (LNM) and central lymph node metastasis (CLNM) in papillary thyroid cancer (PTC). MATERIALS AND METHODS: A total of 346 patients with PTC were enrolled and allocated to training (242) and validation (104) sets. Radiomics features were extracted from arterial and venous phase iodine maps, respectively. Aggregated machine-learning strategy was applied for features selection and construction of 2 radiomics scores (LN rad-score; CLN rad-score). Logistic regression model was employed to establish two radiomics nomograms (nomogram 1: predicting LNM; nomogram 2: predicting CLNM) after incorporating LN or CLN rad-score with clinical predictors. Nomograms performance was determined by discrimination, calibration and clinical usefulness. RESULTS: Nomogram 1 incorporated LN rad-score, age (categorized by 55) and CT reported LN status; Nomogram 2 incorporated CLN rad-score, capsule contact >25% and CT reported CLN status. 2 nomograms both showed good discrimination and calibration in the training (AUC = 0.847; AUC = 0.837) and validation cohorts (AUC = 0.807; AUC = 0.795). Significant improved AUC, net reclassification index (NRI) and integrated discriminatory improvement (IDI) confirmed additional great predictive value of 2 rad-scores, compared with clinical models without radiomics. Decision curve analysis indicated clinical utility of nomograms. 2 nomograms both demonstrated favorable predictive efficacy in CT reported LN or CLN negative subgroup (AUC = 0.766; AUC = 0.744). CONCLUSION: The presented 2 radiomics nomograms are useful tools for preoperative prediction of LNM and CLNM in PTC.

Long noncoding RNA FER1L4 promotes the malignant processes of papillary thyroid cancer by targeting the miR-612/ Cadherin 4 axis.

BACKGROUND: Long noncoding RNAs (lncRNAs) have emerged as crucial regulators in various cancers. However, the functional roles of most lncRNA in papillary thyroid cancer (PTC) are not detailly understood. This study aims to investigate the biological function and molecular mechanism of lncRNA Fer-1 like family member 4 (FER1L4) in PTC. METHODS: The expression of FER1L4 in PTC was determined via operating quantitative real-time PCR assays. Meanwhile, the clinical significance of FER1L4 in patients with PTC was described. The biological functions of FER1L4 on PTC cells were evaluated by gain and loss of function experiments. Moreover, animal experiments were performed to reveal the effect on tumor growth. Subcellular distribution of FER1L4 was determined by fluorescence in situ hybridization and subcellular localization assays. Luciferase reporter assay and RNA immunoprecipitation assay were applied to define the relationship between FER1L4, miR-612, and Cadherin 4 (CDH4). RESULTS: Upregulated expression of FER1L4 in PTC tissues was positively correlated with lymph node metastasis (P = 0.020), extrathyroidal extension (P = 0.013) and advanced TNM stages (P = 0.013). In addition, knockdown of FER1L4 suppressed PTC cell proliferation, migration, and invasion, whereas ectopic expression of FER1L4 inversely promoted these processes. Mechanistically, FER1L4 could competitively bind with miR-612 to prevent the degradation of its target gene CDH4. This condition was further confirmed in the rescue assays. CONCLUSIONS: This study first demonstrates FER1L4 plays an oncogenic role in PTC via a FER1L4-miR-612-CDH4 axis and may provide new therapeutic and diagnostic targets for PTC.

Feasibility study of using virtual non-contrast images derived from dual-energy CT to replace true non-contrast images in patients diagnosed with papillary thyroid carcinoma.

OBJECTIVE: To assess the feasibility of using virtual non-contrast (VNC) images derived from dual-energy computed tomography (DECT) to replace true non-contrast (TNC) images of papillary thyroid carcinoma (PTC) patients. METHODS: Images of 96 PTC patients were retrospectively analyzed. TNC images were acquired under the single-energy mode of DECT after the plain scanning. The arterial and venous phase VNC (VNC-a and VNC-v) images were generated by the post-processing algorithm from the arterial phase and venous phase of contrast-enhanced CT images, respectively. Mean attenuation values, image noise, number and length of calcification were measured. Radiation dose was also calculated. Last, subjective score of image quality was evaluated by a 5-point scale. RESULTS: Signal-to-noise ratio (SNR) of each tissue in TNC images is significantly higher than that of VNC images (p<0.050). Contrast-to-noise ratio (CNR) of fat, muscle, thyroid nodules and internal carotid artery in TNC images is significantly higher than that of VNC images, while CNR in TNC images is lower for cervical vertebra (p<0.001). Calcification is detected on TNC images of 44 patients, while it is omitted on VNC images of 14 patients (31.8%). The subjective score of TNC images is higher than VNC images (p<0.001). The effective dose reduction is 47.6% by avoiding plain scanning. CONCLUSIONS: Considering the different attenuation value, SNR, CNR and especially reduced detection rate of calcification, we deem that VNC images cannot be directly used to replace TNC images in PTC patients, despite the reduced radiation dose.

The direct miR-874-3p-target FAM84A promotes tumor development in papillary thyroid cancer.

With the improvement in diagnostic technology, the incidence of thyroid cancer (TC) is on the rise. Papillary thyroid carcinoma (PTC) is the most common pathological type of thyroid cancer; therefore, it is important to explore some valuable molecular targets to improve the treatment and prognosis of PTC. Studies have shown that family with sequence similarity 84, member A (FAM84A) is involved in the development of various tumors. However, the role of FAM84A in PTC remains unknown. Herein, we explored the biological function and specific molecular mechanism of FAM84A in PTC. Results indicated that FAM84A was upregulated in PTC tissues and cells. In addition, patients with higher FAM84A expression tended to possess larger tumor size, higher lymph node metastasis rate, and advanced TNM stage. Further studies indicated that downregulation of FAM84A could inhibit the development of PTC in vitro and in vivo by repressing the epithelial-mesenchymal transition (EMT) and Wnt/ß-catenin signaling pathway. Moreover, FAM84A was confirmed to be negatively regulated by tumor suppressor miR-874-3p. In conclusion, our findings suggest that FAM84A may act as a potential diagnostic and therapeutic target for PTC.

Management of Sonographically Suspicious Thyroid Nodules 1 cm or Smaller and Candidacy for Active Surveillance: Experience of a Tertiary Center in China.

OBJECTIVE: Our objective was to investigate the management of patients with asymptomatic suspicious thyroid nodules ≤1 cm. METHODS: We retrospectively reviewed medical records of patients with sonographically suspicious thyroid nodules ≤1 cm and without distant metastases, suspicious lymph node metastasis (LNM), or extrathyroidal extension (ETE). RESULTS: Of the 386 enrolled patients, 174 (45.1%) had immediate surgery (IS), while 212 (54.9%) underwent active surveillance (AS). In the IS group, 166 (95.4%) patients were confirmed as having papillary thyroid microcarcinoma. LNM and ETE were observed in 24.7% and 2.4% cases, respectively. In the AS group, nodule size increased by ≥3 mm in 11 (5.2%) patients and 39 (18.4%) had a >50% increase in nodule volume after a median follow-up of 12 months. Nodules with smaller volume at diagnosis were more likely to increase in volume later. Newly suspicious LNM was detected in 23 (10.8%) patients. Delayed surgery (DS) was performed in 101 patients, with 27 showing disease progression. ETE and LNM were detected in 3% and 36%, respectively, of patients with papillary thyroid microcarcinoma. Compared with IS, tumors in the DS group more frequently showed lateral LNM and capsular invasion (P < .05). No patient had recurrence or died of thyroid cancer during postoperative follow-up (median 26 [4-60] months). CONCLUSIONS: IS or DS of patients with asymptomatic suspicious thyroid nodules ≤1 cm was relatively high in China. The inertia of low-risk nodules and the effectiveness of DS for those that progressed make AS a feasible strategy.

Epidemiological characteristics of four common respiratory viral infections in children.

BACKGROUND: Viruses are the main infectious agents of acute respiratory infections in children. We aim to describe the epidemiological characteristics of viral pathogens of acute respiratory tract infections in outpatient children. METHODS: From April 2018 to March 2019, the results of viral detection using oral pharyngeal swabs from 103,210 children with acute respiratory tract infection in the outpatient department of the Children's Hospital, Zhejiang University School of Medicine, were retrospectively analyzed. Viral antigens, including adenovirus (ADV), influenza A (FLUA), influenza B (FLUB) and respiratory syncytial virus (RSV), were detected by the colloidal gold method. RESULTS: At least one virus was detected in 38,355 cases; the positivity rate was 37.2%. A total of 1910 cases of mixed infection with two or more viruses were detected, and the positivity rate of multiple infection was 1.9%. The ADV positivity rate was highest in the 3-6-year-old group (18.7%), the FLUA positivity rate was highest in the > 6-year-old group (21.6%), the FLUB positivity rate was highest in the > 6-year-old group (6.6%), and the RSV positivity rate was highest in the < 1-year-old group (10.6%). There was a significant difference in the positivity rate of viral infection among different age groups (χ2 = 1280.7, P < 0.001). The rate of positive viral infection was highest in winter (47.1%). The ADV infection rate was highest in spring (18.2%). The rates of FLUA and FLUB positivity were highest in winter (28.8% and 3.6%, respectively). The rate of RSV positivity was highest in autumn (17.4%). The rate of positive viral infection in different seasons was significantly different (χ2 = 6459.1, P < 0.001). CONCLUSIONS: Viral infection rates in children differ for different ages and seasons. The positivity rate of ADV is highest in the preschool period and that of RSV is highest in infants; that of FLU increases with age. The total positive rate of viral infection in different seasons is highest in winter, as is the rate of FLU positivity.

Texture analysis of dual-phase contrast-enhanced CT in the diagnosis of cervical lymph node metastasis in patients with papillary thyroid cancer.

BACKGROUND: Computed tomography texture analysis (CTTA) provides objective and quantitative information regarding tumor heterogeneity beyond visual inspection. However, no study has yet used CTTA to differentiate metastatic from non-metastatic cervical lymph node in patients with papillary thyroid cancer (PTC). PURPOSE: To evaluate the value of texture analysis of dual-phase contrast-enhanced CT images in diagnosing cervical lymph node metastasis in patients with PTC. MATERIAL AND METHODS: Metastatic (n = 27) and non-metastatic (n = 32) cervical lymph nodes were analyzed retrospectively. Texture analyses were performed on both arterial (A) and venous (V) phase CT images. Texture parameters, including mean gray-level intensity, skewness, kurtosis, entropy, and uniformity, were obtained and compared between groups. Receiver operating characteristic (ROC) curves analyses and multivariate logistic regression analysis were used in our study. RESULTS: Metastatic lymph nodes showed significantly higher A-mean gray-level intensity, A-entropy, and lower A-kurtosis and V-kurtosis (all P < 0.001) than non-metastatic mimics. The ROC curve analyses indicated that A-kurtosis demonstrated an optimal diagnostic area under the curve (AUC; 0.884) and specificity (92.59%), while the A-mean gray-level intensity showed optimal diagnostic sensitivity (90.62%). Multivariate logistic regression analysis showed that A-mean gray-level intensity (P = 0.006, odds ratio [OR] = 24.297) and V-kurtosis (P = 0.014, OR = 19.651) were the independent predictor for metastatic cervical lymph node. CONCLUSION: Dual-phase contrast-enhanced CCTA-especially A-mean gray-level intensity and V-kurtosis-may have the potential to diagnose metastatic cervical lymph node in patients with PTC.

Investigation of anxiety levels of 1637 healthcare workers during the epidemic of COVID-19.

OBJECTIVE: This study aimed to investigate the anxiety levels of healthcare workers and to provide guidance on potential accurate social and psychological interventions for healthcare workers during the epidemic of COVID-19 in Zhejiang Province, China. METHODS: Healthcare workers from five hospitals in Zhejiang Province were randomly selected into this study. Zung Self-Assessment Scale for Anxiety (SAS) was used to evaluate the anxiety status of the included 1637 healthcare workers. RESULTS: The total anxiety score of healthcare workers in Zhejiang Province was 30.85 ± 6.89. The univariate analysis showed that the anxiety level of healthcare workers was related to gender, education, occupation, physical condition, job risk coefficient, and with family members on the first-line combating COVID-19 (P <0.05). The multivariate analysis showed that physical condition and job risk coefficient were predictors of anxiety levels of healthcare workers. CONCLUSIONS: During the epidemic of COVID-19, 1637 healthcare workers generally had an increased tendency to have anxiety. Individualized assessment of the anxiety level of healthcare workers should be provided, and different interventions should be given based on the evaluation results.

[Application of three-in-one intelligent screening in outpatient department of children's hospital during COVID-19 epidemic].

OBJECTIVE: To evaluate the application of three-in-one intelligent screening in outpatient pre-inspection in children's hospital. METHODS: We randomly enrolled 100 children pre-screened by traditional method in the outpatient department of Children's Hospital of Zhejiang University from February 6th to 16th, 2020, and another 100 children by the intelligent three-in-one mode from February 17th to 27th, 2020. The traditional triage was conducted by nurses based on face-to-face, one-by-one interview of the epidemiological history and consultation department, and the temperature was measured before manual triage. The intelligent three-in-one model combined online rapid pre-inspection and triage, on-site manual confirmation, as well as synchronized online health education system. For on-line registered patients, the system automatically sent the COVID-19 epidemiological pre-screening triage questionnaire one hour before the appointment, requiring parents to complete and submit online before arriving at the hospital. The on-site registered patients were controlled at 100 m away from the hospital entrance. The nurses guided the parents to scan the QR code and fill in the COVID-19 epidemiological pre-examination triage questionnaire. At the entrance of the hospital, the nurse checked the guidance sheet and took the temperature again. The children with red guidance sheet were checked again and confirmed by pre-examination nurses, and accompanied to the isolation clinic through COVID-19 patients-only entrance. The children with yellow guidance sheet were guided to fever clinic. The children with green guidance sheet could go with their parents to the designated area, and then went to the corresponding consultation area. Health education was carried out throughout the treatment, and the system automatically posted the corresponding outpatient instructions and education courses. Parents would read the courses on their mobile phones and counsel online. The time of pre-examination and the coincidence rate of triage were compared between the two groups. RESULTS: The three-in-one intelligent pre-inspection mode took an average of (25.6±8.0) s for each child, which was significantly shorter than the traditional pre-inspection mode (74.8±36.4) s (t=13.182, P<0.01). The triage coincidence rate of the intelligent pre-inspection model was 98%, which was similar to that of traditional model (97%, χ2=0.251, P>0.05). CONCLUSIONS: The three-in-one intelligent pre-inspection model can effectively shorten the patient pre-check time, with similar triage coincidence rate to traditional model.

Radiomics analysis of dual-energy CT-derived iodine maps for diagnosing metastatic cervical lymph nodes in patients with papillary thyroid cancer.

OBJECTIVE: To investigate the value of radiomics analysis of dual-energy computed tomography (DECT)-derived iodine maps for preoperative diagnosing cervical lymph nodes (LNs) metastasis in patients with papillary thyroid cancer (PTC). METHODS: Two hundred and fifty-five LNs (143 non-metastatic and 112 metastatic) were enrolled and allocated to training and validation sets (7:3 ratio). Radiomics features were extracted from arterial and venous phase iodine maps, respectively. Radiomics signature was constructed based on reproducible features using the least absolute shrinkage and selection operator (LASSO) logistic regression algorithm with 10-fold cross-validation. Logistic regression modeling was employed to build models based on CT image features (model 1), radiomics signature (model 2), and the combined (model 3). A nomogram was plotted for the combined model and decision curve analysis was applied for clinical use. Diagnostic performance was assessed and compared. Internal validation was performed on an independent set containing 78 LNs. RESULTS: Model 3 showed optimal diagnostic performance in both training (AUC = 0.933) and validation set (AUC = 0.895), followed by model 2 (training set, AUC = 0.910; validation set, AUC = 0.847). Both these two models outperformed model 1 in both training (AUC = 0.763) (p < 0.05) and validation set (AUC = 0.728) (p < 0.05). CONCLUSION: Radiomics analysis of DECT-derived iodine maps showed better diagnostic performance than qualitative evaluation of CT image features in preoperative diagnosing cervical LN metastasis in PTC patients. Radiomics signature integrated with CT image features can serve as a promising imaging biomarker for the differentiation. KEY POINTS: • Conventional CT image features have limited value for the diagnosis of metastatic LNs in PTC patients. • Radiomics analysis of dual-energy CT-derived iodine maps significantly outperformed qualitative CT image features in differentiating metastatic from non-metastatic LNs. • Radiomics signature integrated with qualitative CT image features can serve as a useful tool in judging LNs status, thus aiding clinical decision-making.

Ultrasonic scalpel with knot tying protects parathyroid function for total thyroidectomy with central neck dissection.

BACKGROUND: Hypoparathyroidism might be due to collateral thermal injury induced by harmonic scalpel (HS) after total thyroidectomy (TT) with central neck dissection (CND). The current study aimed to evaluate whether a clamp and tie (CAT) technique used to preserve parathyroid glands in situ could reduce damage to parathyroid function in HS patients undergoing TT plus unilateral or bilateral CND. METHODS: Medical records of 537 HS-operated patients with papillary thyroid carcinoma (PTC) undergoing TT plus unilateral or bilateral CND were retrospectively evaluated. Patients were divided into HS and HS-CAT groups based on whether the CAT techniques were used near parathyroid glands. Patients' characteristics such as age, sex, tumour size, operative time, pre- and post-operative levels of parathyroid hormone (PTH) and complications were analysed. RESULTS: For patients undergoing TT plus unilateral CND, operative time was significantly shorter (P<0.001), but there were significantly higher incidences of transient hypoparathyroidism (P=0.002) on postoperative day 1 and incidental parathyroidectomy (P=0.036) in the HS group. There was no significant difference in permanent hypoparathyroidism. For patients undergoing TT plus bilateral CND, a significantly shorter operative time and sharper postoperative PTH decline was found in the HS group (P=0.029). However, there were no significant differences regarding incidences of incidental parathyroidectomy, transient or permanent hypoparathyroidism. CONCLUSIONS: The CAT technique applied near parathyroid glands was a practical surgical application for decreasing the incidence of postoperative transient hypoparathyroidism for TT plus unilateral CND.