A Cas12a-based fluorescent microfluidic system for rapid on-site human papillomavirus diagnostics.

Persistent infection with human papillomavirus (HPV) is the leading cause of cervical cancer, and early diagnosis is crucial for clinical management. However, the easy and rapid on-site diagnostic for HPV genotyping remains challenging. Here, we develop a Cas12a-based fluorescent microfluidic detection system for diagnosing six HPV subtypes (HPV6, HPV11, HPV16, HPV18, HPV31, and HPV33). A panel of crRNAs and recombinase polymerase amplification (RPA) primers targeting the HPV L1 gene was screened for sensitive and specific detection. Furthermore, a one-pot RPA reaction was developed to amplify the six HPV subtypes without cross-reactivity. For on-site detection, we integrated the RPA-Cas12a detection into a microfluidic device, enabling the detection of processed clinical samples within 35 minutes. The assay was validated using 112 clinical swab samples and obtained consistent results with the qPCR assay, with a concordance rate of 99.1%. Overall, our diagnostic method offers a rapid, sensitive, and easy-to-use on-site assay for detecting HPV genotypes and holds promise for improving cervical cancer screening and prevention. KEY POINTS: • The Cas12a-based fluorescent microfluidic detection system for the diagnosis of six HPV subtypes. • A one-pot RPA reaction for amplifying the six HPV subtypes without cross-reactivity. • The RPA-Cas12a-microfluidic system provides results within 35 minutes for on-site detection.

Development and Validation of the Missed Nursing Care Tool for Pre-Operative Patients with Lung Cancer in China.

Introduction: Current research suggests that missed nursing care is widespread in preoperative lung cancer patients in China, and preoperative airway management nursing care for lung cancer patients is not standardized. Missed nursing care for preoperative lung cancer patients, on the other hand, is rarely investigated, particularly from the patient's perspective. This study aimed to develop and validate the MISSCARE Survey for pre-operative patients with lung cancer in China. Methods: This study generated the preliminary draft of the MISSCARE Survey - Lung Cancer Patient (MS-LCP) and tested its reliability and stability through three rounds of lung cancer resection (494, 50, and 309 cases, respectively). 20 patients and 6 experts determined the face and content validity. EFA and CFA assessed construct and convergent validity. Internal consistency, including Cronbach's alpha, Spearman-Brown reliability, and re-test reliability, was also examined. Results: The scale contained 15 items, including specific care, communicative care, timely care, and basic care. KMO was 0.932 (> 0.6), and Bartlett's Test of Sphericity showed P = 0.000 (<0.05). The attribution factor's item loads ranged from 0.765 to 0.853, accounting for 82.20% of the variation. The scale's Cronbach's alpha, Spearman-Brown, and retest reliability were 0.945, 0.879, and 0.824. CFA showed goodness of fit (RMSEA = 0.021, χ2/df = 1.138, GFI = 0.900, AGFI = 0.945, CFI = 0.996, NFI = 0.967, IFI = 0.996). For each dimension, AVE ranged from 0.555 to 0.717 (>0.50) and CR from 0.861 to 0.904 (>0.70). Conclusion: The MS-LCP was reliable and valid in this study, making it appropriate for the Chinese lung cancer patient population. This tool is more objective in its presentation of missed nursing care, assisting nursing staff in optimizing nursing procedures before surgery, increasing the implementation of higher-quality tumor care, and promoting the recovery of lung cancer surgery patients.

Analysis of sternal healing after median sternotomy in low risk patients at midterm follow-up: retrospective cohort study from two centres.

BACKGROUND: For low risk patients undergoing median sternotomies, no midterm follow-up studies involving sternal healing have been conducted. In this study we evaluated sternal healing in low risk patients by chest CT scan and the risk factors associated with poor healing were analyzed. METHODS: Patients who underwent sternal median incision heart surgery from September 2014 to March 2015 were recruited. The clinical information of these patients during hospitalization was collected, and the CT scan data were submitted to the two chief physicians of the Radiology Department for radiographical sternal healing score determination. Based on the method of wound closure, the patients were divided into sternum plate (Plates) and wire groups (Wires). RESULTS: Forty-four patients were recruited. The mean CT examination time was 17.27 ± 2.30 months postoperatively. Twenty-nine (65.9%) patients met the criteria for radiographic sternal healing. Three segments, including the aortopulmonary window, the main pulmonary artery, and the aortic root, had healed less in comparison to the manubrium segment. Compared to patients in whom 6-7 metal wires were used for sternal closure, healing of the lower sternum was worse in patients in whom five wires were used, but the difference in healing was not statistically significant. Univariate analysis of sternal healing showed that patient age was a risk factor for sternal non-healing. When the patient age was > 45 years, the predicted risk of radiographic sternal non-union was 1.833 (95% CI: 1.343-2.503). CONCLUSIONS: At the mid-term follow-up, 65.9% of patients undergoing median sternotomies demonstrated radiographic sternal healing. Age, but not closure device, was a risk factor for sternal non-healing in low risk patients. Use of more wires had a positive impact on sternal healing. TRIAL REGISTRATION: researchregistry4918, registered 28 May 2019, retrospectively registered.

PCA-Based Multiple-Trait GWAS Analysis: A Powerful Model for Exploring Pleiotropy.

Principal component analysis (PCA) is a potential approach that can be applied in multiple-trait genome-wide association studies (GWAS) to explore pleiotropy, as well as increase the power of quantitative trait loci (QTL) detection. In this study, the relationship of test single nucleotide polymorphisms (SNPs) was determined between single-trait GWAS and PCA-based GWAS. We found that the estimated pleiotropic quantitative trait nucleotides (QTNs) ß * ^ were in most cases larger than the single-trait model estimations ( ß 1 ^ and ß 2 ^ ). Analysis using the simulated data showed that PCA-based multiple-trait GWAS has improved statistical power for detecting QTL compared to single-trait GWAS. For the minor allele frequency (MAF), when the MAF of QTNs was greater than 0.2, the PCA-based model had a significant advantage in detecting the pleiotropic QTNs, but when its MAF was reduced from 0.2 to 0, the advantage began to disappear. In addition, as the linkage disequilibrium (LD) of the pleiotropic QTNs decreased, its detection ability declined in the co-localization effect model. Furthermore, on the real data of 1141 Simmental cattle, we applied the PCA model to the multiple-trait GWAS analysis and identified a QTL that was consistent with a candidate gene, MCHR2, which was associated with presoma muscle development in cattle. In summary, PCA-based multiple-trait GWAS is an efficient model for exploring pleiotropic QTNs in quantitative traits.

Impedimetric determination of Staphylococcal enterotoxin B using electrochemical switching with DNA triangular pyramid frustum nanostructure.

An electrochemical switching strategy is presented for the sensitive determination of Staphylococcus enterotoxin B (SEB). It is based on the use of DNA triangular pyramid frustum nanostructure (TPFDNA) consisting of (a) three thiolated probes, (b) one auxiliary probe, and (c) an aptamer against SEB. The TPFDNA was assembled on the gold electrode, with the SEB aptamer designed on top of the TPFDNA. The electron transfer to hexacyanoferrate acting as an electrochemical probe is strongly inhibited in the TPFDNA-modified electrode. This is assumed to be due to the formation of a 3D TPFDNA structure that limits access of hexacyanoferrate to the electrode. Therefore, the Faradaic impedance is large. However, in the presence of SEB, it will bind to the aptamer and dehybridize the hybrid formed between aptamer and its complementary sequence. As a result, the TPFDNA nanostructure changes to an equilateral triangle DNA nanostructure. This results in a more efficient electron transfer and a smaller Faradaic impedance. The method has a detection limit of 0.17 ng mL-1 of SEB (at an S/N of 3) and a dynamic range that covers the 0.2-1000 ng mL-1 concentration range. The applicability and reliability of the method was demonstrated by anayzing (spiked) milk samples, and the results were compared to those obtained with an ELISA kit. The relative standard deviations between the two methods range between -6.59 and 9.33%. Graphical abstract An electrochemical switching strategy is presented for the sensitive detection of Staphylococcus enterotoxin B based on 3D DNA structure conversion of nanostructure from triangular pyramid frustum to equilateral triangle.

Rapid diagnosis of sepsis with TaqMan-Based multiplex real-time PCR.

BACKGROUND: The survival rate of septic patients mainly depends on a rapid and reliable diagnosis. A rapid, broad range, specific and sensitive quantitative diagnostic test is the urgent need. Thus, we developed a TaqMan-Based Multiplex real-time PCR assays to identify bloodstream pathogens within a few hours. METHODS: Primers and TaqMan probes were designed to be complementary to conserved regions in the 16S rDNA gene of different kinds of bacteria. To evaluate accurately, sensitively, and specifically, the known bacteria samples (Standard strains, whole blood samples) are determined by TaqMan-Based Multiplex real-time PCR. In addition, 30 blood samples taken from patients with clinical symptoms of sepsis were tested by TaqMan-Based Multiplex real-time PCR and blood culture. RESULTS: The mean frequency of positive for Multiplex real-time PCR was 96% at a concentration of 100 CFU/mL, and it was 100% at a concentration greater than 1000 CFU/mL. All the known blood samples and Standard strains were detected positively by TaqMan-Based Multiplex PCR, no PCR products were detected when DNAs from other bacterium were used in the multiplex assay. Among the 30 patients with clinical symptoms of sepsis, 18 patients were confirmed positive by Multiplex real-time PCR and seven patients were confirmed positive by blood culture. CONCLUSION: TaqMan-Based Multiplex real-time PCR assay with highly sensitivity, specificity and broad detection range, is a rapid and accurate method in the detection of bacterial pathogens of sepsis and should have a promising usage in the diagnosis of sepsis.

Genome-wide analysis reveals differential selection involved with copy number variation in diverse Chinese Cattle.

Copy number variations (CNVs) are defined as deletions, insertions, and duplications between two individuals of a species. To investigate the diversity and population-genetic properties of CNVs and their diverse selection patterns, we performed a genome-wide CNV analysis using high density SNP array in Chinese native cattle. In this study, we detected a total of 13,225 CNV events and 3,356 CNV regions (CNVRs), overlapping with 1,522 annotated genes. Among them, approximately 71.43 Mb of novel CNVRs were detected in the Chinese cattle population for the first time, representing the unique genomic resources in cattle. A new V i statistic was proposed to estimate the region-specific divergence in CNVR for each group based on unbiased estimates of pairwise V ST . We obtained 12 and 62 candidate CNVRs at the top 1% and top 5% of genome-wide V i value thresholds for each of four groups (North, Northwest, Southwest and South). Moreover, we identified many lineage-differentiated CNV genes across four groups, which were associated with several important molecular functions and biological processes, including metabolic process, response to stimulus, immune system, and others. Our findings provide some insights into understanding lineage-differentiated CNVs under divergent selection in the Chinese native cattle.

Associations Between Inflammatory Markers, Hemostatic Markers, and Microvascular Complications in 182 Chinese Patients With Type 2 Diabetes Mellitus.

OBJECTIVE: To examine the associations between inflammatory markers, coagulation and fibrinolysis parameters, and microvascular complications in 182 Chinese patients with type 2 diabetes mellitus (T2DM) who sought treatment at a large hospital in Zhejiang province, China. METHODS: We investigated the relationships of blood inflammatory markers with hemostatic markers in 87 patients with T2DM who did not have complications and 95 patients with T2DM who had microvascular complications. RESULTS: C-reactive protein (CRP) and interleukin-6 (IL-6) were significantly correlated with fibrinogen, thrombin-antithrombin III complex (TAT III), plasminogen activator inhibitor-1 (PAI-1), von Willebrand factor (vWF), and coagulation factors (F) VII in patients with T2DM who had microvascular complications (P <.05). Based on logistic regression analysis, the highest-tertile groups of fibrinogen, FVII, and FVIII, corresponded to a greater risk of high CRP, whereas risk of high IL-6 was significantly greater in the groups with highest-tertile values for fibrinogen, FVII, TAT III, PAI-1, and activated protein C (APC). CONCLUSIONS: Elevated levels of CRP and IL-6 might be associated with increased coagulability and a tendency towards thrombus formation in patients with T2DM who have microvascular complications.

Relationship between Serum Sialic Acid and Hemostatic Markers in Patients with Type 2 Diabetes Mellitus.

BACKGROUND: Serum sialic acid (SA) is an acute phase response marker. Additionally, it is an independent risk factor for atherosclerotic disease and is higher in patients with type 2 diabetes mellitus (T2DM). The objective of this study was to determine the association of SA with blood coagulation and fibrinolysis in T2DM patients. METHODS: The levels of serum SA and blood coagulation and fibrinolysis markers were measured in patients with T2DM. The associations of SA with hemostatic markers and other variables were assessed. RESULTS: There were significant correlations between SA and fibrinogen, D-dimer, factor (F) IX, and platelet (PLT) that were independent of smoking, body mass index (BMI), waist-to-hip ratio, hemoglobin A(1c) (HbA1c), fasting plasma glucose (FPG), triglycerides (TG), total cholesterol (TC) to high-density lipoprotein cholesterol (HDL-C) ratio, and antithrombotic therapy history. Based on logistic regression analyses, the highest tertile groups of fibrinogen, D-dimer, F VIII, F IX, and PLT showed a significantly increased risk of high SA compared with the lowest tertiles after adjusting for age, gender, and antithrombotic therapy history. SA also significantly correlated with duration of diabetes, BMI, FPG, TG, low-density lipoprotein cholesterol (LDL-c), TC-to-HDL-C ratio, and albumin-to-creatinine ratio (ACR). CONCLUSIONS: Elevated serum SA levels are associated with increased coagulability and higher risk of thrombus formation in T2DM patients.

Intravenous leiomyomatosis of the uterus with extension to the right heart.

A 42-year-old woman admitted with debilitation and engorgement both lower extremities. Transthoracic two-dimensional echocardiography, abdominal ultrasound and computerized tomography revealed a lobulated pelvic mass, a mass within right internal iliac vein, both common iliac vein, as well as the inferior vena cava, extending into the right atrium. In addition, echocardiography and abdominal ultrasound showed the tumor of right atrium and inferior vena cave has no stalk and has well-demarcated borders with the wall of right atrium and inferior vena cave. Hence, the presumptive diagnosis of IVL was made by echocardiography and abdominal ultrasound and the presumptive diagnosis of sarcoma with invasion in right internal iliac vein, both common iliac vein, the inferior vena cava, as well as the right atrium was made by multi-detector-row computerized tomography. The patient underwent a one-stage combined multidisciplinary thoraco-abdominal operation under general anaesthetic. Subsequently the pathologic report confirmed IVL.