Computed Tomography Image Features under Deep Learning Algorithm Applied in Staging Diagnosis of Bladder Cancer and Detection on Ceramide Glycosylation

The research is aimed at investigating computed tomography (CT) image based on deep learning algorithm and the application value of ceramide glycosylation in diagnosing bladder cancer. The images of ordinary CT detection were improved. In this study, 60 bladder cancer patients were selected and performed with ordinary CT detection, and the detection results were processed by CT based on deep learning algorithms and compared with pathological diagnosis. In addition, Western Blot technology was used to detect the expression of glucose ceramide synthase (GCS) in the cell membrane of tumor tissues and normal tissues of bladder. The comparison results found that, in simple CT clinical staging, the coincidence rates of T1 stage, T2a stage, T2b stage, T3 stage, and T4 stage were 28.56%, 62.51%, 78.94%, 84.61%, and 74.99%, respectively; and the total coincidence rate of CT clinical staging was 63.32%, which was greatly different from the clinical staging of pathological diagnosis ( P < 0.05 ). In the clinical staging of algorithm-based CT test results, the coincidence rates of T1 stage and T2a stage were 50.01% and 91.65%, respectively; and those of T2b stage, T3 stage, and T4 stage were 100.00%; and the total coincidence rate was 96.69%, which was not obviously different from the clinical staging of pathological diagnosis ( P > 0.05 ). Therefore, it could be concluded that the algorithm-based CT detection results were more accurate, and the use of CT scans based on deep learning algorithms in the preoperative staging and clinical treatment of bladder cancer showed reliable guiding significance and clinical value. In addition, it was found that the expression level of GCS in normal bladder tissues was much lower than that in bladder cancer tissues. This indicated that the changes in GCS were closely related to the development and prognosis of bladder cancer. Therefore, it was believed that GCS may be an effective target for the treatment of bladder cancer in the future, and further research was needed for specific conditions.

Impact of Short-Incubation MALDI-TOF MS on Rapid Identification of Pathogens and Antibiotic Therapy

Background: It can be a critical point for reducing pathogen identification time and accurate antibiotic treatment for patients with blood circulation infection since it causes high mortality. Objective: The objectives of this study were to evaluate the time differences between conventional identification and MALDI-TOF conventional identification and short-incubation MALDI-TOF identification for positive blood cultures, and to explore the impact of short-incubation MALDI-TOF identification on empirical antibiotic therapy. Methods: Positive blood cultures were collected in our hospital from 2017 to 2019, clinical data were collected from the medical records, which were analyzed retrospectively to determine the empirical antibiotic therapy. Results: Compared with the conventional identification method, the short-incubation MALDI-TOF identification time to initial identification of Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecium, and E. faecalis decreased by 22.28 h, 22 h, 23.59 h, 23.63 h, 22.63 h, 23.92 h, and 21.59 h, respectively (P < 0.05). The time to final reporting was decreased by 48.85 h, 47.99 h, 55.40 h, 51.07 h, 49.60 h, 51.78h, and 51.73h, respectively (P < 0.05). However, the antimicrobial susceptibility test time of E. coli, A. baumannii, and S. aureus increased to 2.02 h, 2.19 h, and 3.86 h, respectively (P < 0.05). The coincidence rate of antimicrobial susceptibility was 98.48% between short-incubation MALDI-TOF identification and conventional identification method of all Gram-negative bacilli, and there were no extremely major errors or major errors. The coincidence rate of antimicrobial susceptibility of Gram-positive cocci was 99.53%, one strain of E. faecium and S. aureus had major errors. Patients received earlier correct empirical antibiotic 19.89 h earlier by short-incubation MALDI-TOF identification than the conventional identification method (P < 0.001). Conclusions: The short-incubation MALDI-TOF identification significantly shortens the pathogen identification time and the final report time, it is a reliable method for rapid identification of positive blood cultures; the results of antimicrobial susceptibility are highly consistent, which significantly lead to earlier appropriate empirical therapy of bacteremia.

Development of a reverse transcription recombinase-aided amplification assay for detection of Getah virus

GETV, an arbo-borne zoonotic virus of the genus Alphavirus, which causes diarrhea and reproduction disorders in swine, lead to serious economic losses to the swine industry in China. At present, the existing methods for GETV detection are time-consuming and low sensitivity, so, a rapid, accurate and sensitive GETV detection method is urgently needed. In this study, a fluorescent reverse transcription recombinase-assisted amplification method (RT-RAA) was successfully established for the rapid detection of GETV. The sensitivity of this method to GETV was 8 copies/reaction and 20 TCID50/reaction. No cross-reaction with other viruses. A total of 118 samples were prepared for GETV detection using fluorescent RT-RAA and SYBR Green I RT-qPCR, the coincidence rate of the two methods was 100%. The results suggest that the RT-RAA method is rapid, sensitive and specific for GETV detection and can be applied in the clinical.

A Rapid and Sensitive Detection Method for Pseudomonas aeruginosa Using Visualized Recombinase Polymerase Amplification and Lateral Flow Strip Technology

Pseudomonas aeruginosa is a common opportunistic pathogen that causes acute nosocomial necrotizing pneumonia and is the predominant source of chronic lung infections in patients with the genetic disorder cystic fibrosis. Early diagnosis in infected patients and monitoring P. aeruginosa contamination is therefore of great importance in controlling disease spread and development with timely drugs intervention treatment and cut off infection source. Traditional culture-biochemical methods are time consuming and highly dependent on technicians and expensive instruments. To address these challenges, the present study aimed to develop a rapid, sensitive, and specific, on-site detection method for P. aeruginosa based on recombinase polymerase amplification (RPA) combined with lateral flow strip (LFS) technology. The experimental process included screening and modification of primer and probe sets targeting the unique virulence gene elastase B (lasB); specificity detection in 29 strains of P. aeruginosa and 23 closely-related pathogenic bacteria; sensitivity measurements with gradient-diluted P. aeruginosa genomic DNA and probit regression analysis; and clinical application evaluation using 574 patients samples and calculating coincidence rate and kappa index value in comparison with the culture-biochemical method. The P. aeruginosa RPA-LFS assay could complete the amplification process at 37°C constant temperature within 30 min and results could be visualized by the naked eye within 10 min on LFS. The assay displayed high sensitivity with a limit of detection of 3.05 CFU/reaction. It also demonstrated high specificity by showing no cross reaction with other pathogenic bacteria, and rapidness by being completed in less than an hour. Furthermore, when used with clinical samples, the assay had a coincidence rate of 98.26% with the culture-biochemical method and a kappa index value of 0.9433. These data indicate that the RPA-LFS assay represents a major improvement for P. aeruginosa detection, especially in resource-limited areas.

Clinical value of contrast-enhanced computed tomography (CECT) combined with contrast-enhanced ultrasound (CEUS) for characterization and diagnosis of small nodular lesions in liver

Objectives: To explore the clinical value of contrast-enhanced computed tomography (CECT) combined with contrast-enhanced ultrasound (CEUS) for characterization and diagnosis of small nodular lesions in the liver and investigate the association between such small nodular lesions and the degree of tumor differentiation. Methods: Combined imaging modalities were performed on 120 patients who were admitted by Linyi Maternal and Child Health hospital from December 2018 to December 2020 and diagnosed with hepatic nodular lesions. The CT scans were interpreted by two senior imageologists while the ultrasound scans were analyzed by two senior sonographers. A comparative analysis was carried out on different scan modes and the postoperative or post-puncture pathological results using the t-test, the χ2 test, and the Pearson’s correlation analysis. Results: Compared to the pathological results, definite diagnoses of 55 malignant cases were made using CECT alone, with the coincidence rate of 78.6%; CECT combined with CEUS formed correct diagnoses in 64 cases, and the coincidence rate was up to 91.4%. The difference between the two scan modes was statistically significant (p= 0.03). Based on pathological diagnosis, seventy out of the 120 cases of small nodular lesions were identified as malignant, while the other 50 cases were benign. The single imaging modality diagnosed 63 malignant and 57 benign nodules, whereas the combined modalities identified 68 malignancies and 52 benign conditions. Compared to CECT as a single imaging modality, the combined modalities showed a higher degree of sensitivity and accuracy, and the difference was statistically significant (sensitivity: p= 0.03; accuracy: p= 0.02); in the malignant cases, the magnitudes of contrast enhancement of CT and ultrasound imaging decreased with an increase in the degree of differentiation, indicating a negative correlation between these factors. Conclusions: CECT combined with CEUS has a higher coincidence rate, greater sensitivity, and better diagnostic accuracy when being used for characterization and diagnosis of small nodular lesions in the liver. A higher degree of tumor differentiation means a decreased magnitude of contrast enhancement and a blurrier boundary, which indicates that CECT and CEUS are complementary to each other in classifying malignant liver nodules. The use of the combined imaging modalities shows clinical value for characterizing small liver nodules and predicting the degree of malignancy. doi: https://doi.org/10.12669/pjms.37.7.4306 How to cite this:Liu J, Bao D, Xu Z, Zhuge X. Clinical value of contrast-enhanced computed tomography (CECT) combined with contrast-enhanced ultrasound (CEUS) for characterization and diagnosis of small nodular lesions in liver. Pak J Med Sci. 2021;37(7):---------. doi: https://doi.org/10.12669/pjms.37.7.4306 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Single Multiple Cross Displacement Amplification for Rapid and Real-Time Detection of Porcine Circovirus 3

Since 2016, a novel porcine circovirus, PCV3, has been infecting pigs, causing significant economic losses to the pig industry. In recent years, the infection rate of PCV3 has been increasing, and thus rapid and accurate detection methods for PCV3 are essential. In this study, we established a novel probe-based single multiple cross displacement amplification (P-S-MCDA) method for PCV3. The method was termed as P-S-MCDA. The P-S-MCDA uses seven primers to amplify the capsid gene, and the assay can be performed at 60°C for 30 min, greatly shortening the reaction time. The results of P-S-MCDA can not only be monitored in real time through fluorescence signals but also be determined by observing the fluorescence of the reaction tubes using a smartphone-based cassette. This method demonstrated good specificity and the same sensitivity as qPCR, with a minimum detection limit of 10 copies. In 139 clinical samples, the coincidence rate with qPCR was 100%. The P-S-MCDA can be widely applied in PCV3 detection in laboratories or in rural areas.

Combination IETA Ultrasonographic Characteristics Simple Scoring Method With Tumor Biomarkers Effectively Improves the Differentiation Ability of Benign and Malignant Lesions in Endometrium and Uterine Cavity

ObjectivesTo evaluate International Endometrial Tumor Analysis (IETA) ultrasonographic characteristics simple scoring method and tumor biomarkers for the diagnosis of uterine cavity and endometrial lesions.MethodsWe classified and scored the normalized description of IETA ultrasonic characteristics, according to IETA expert consensus literature, previous IETA-related research articles, and the previous research experience of this project group. We conducted a retrospective analysis of the ultrasound images of 594 patients enrolled from January 2017 to June 2020, scored them item by item, and finally calculated the total score of each case. Meanwhile, we combined the results of seven tumor biomarkers. The objective was to evaluate the sensitivity, specificity, coincidence rate, and the area under receiver operating characteristic (ROC) curve of IETA ultrasonographic characteristics simple scoring method and tumor biomarkers for benign and malignant uterine cavity or endometrial lesions. The diagnostic efficiency between the combined method and the single method was compared.ResultsA total of 594 cases were confirmed by postoperative pathology or surgery records, including 475 benign lesions and 119 malignant lesions. In the simple ultrasound scoring method, the average score of benign lesions was 3.879 ± 1.279 and that of malignant lesions was 9.676 ± 4.491. If ≥6.5 points was taken as the cutoff value for the judgment of malignant lesions, the sensitivity, specificity, coincidence rate, and the area under receiver operating characteristic (ROC) curve (AUC) were 76.5%, 96.0%, 92.1%, and 0.935, respectively. The difference in tumor antigen 19-9 (CA19-9) and human epididymal protein 4 (HE4) between benign and malignant lesions was statistically significant (all p ≤ 0.01). The other five tumor biomarkers (CA125, CA15-3, SCC-Ag, AFP, and CEA) showed no statistically significant difference in benign and malignant lesions. If the value of CA19-9 ≥13.96 U/ml was taken as cutoff value, the sensitivity, specificity, and coincidence rate of the diagnosis of endometrial benign and malignant lesions were 54.8%, 74.7%, and 70.7%, respectively, and the AUC was 0.620. If the value of HE4 ≥ 39.075 pmol/L was taken as cutoff point, the sensitivity, specificity, coincidence rate, and AUC were 77.4%, 67.9%, 69.8%, and 0.796, respectively. The sensitivity was increased to 97.6% and the AUC was 0.939 when IETA ultrasound characteristics simple scoring method combined CA19-9 and HE4 in parallel test.ConclusionsIn IETA ultrasound characteristics simple scoring method, with ≥6.5 points as the cutoff value, it could quickly and accurately assess the benign and malignant in uterine cavity and endometrial lesions, with high diagnostic value. The diagnostic efficacy of seven tumor biomarkers was all mediocre. Combining with these two methods, the comprehensive diagnosis could improve sensitivity and accuracy and reduce the risk of missed diagnosis.

A Simple Scoring Scale for Predicting the Risk of Diabetic Retinopathy over 50 Years Old in China

Purpose: To construct and evaluate a simple scoring scale for predicting the risk of diabetic retinopathy (DR).Methods: Based on the chronic disease management database of Yancheng City, Jiangsu Province, China, 1896 diagnosed patients over the age of 50 with diabetes were randomly selected and subjected to the self-designed epidemiological questionnaire survey and ocular clinical examination. Single-factor and multi-factor logistic regression analysis was used to screen the relevant influencing factors of DR and then according to the reference value principle, the weights were assigned, and a simple scoring scale was constructed. A receiver operating characteristic curve (ROC) was developed and the accuracy and validity of the simple scale were evaluated.Results: The DR detection rate was 34.8 %. Multivariate analysis showed that family history of diabetes (odds ratio [OR]: 1.322, 95 % confidence interval [CI]: 1.012-1.727), diabetes treatment method (OR: 2.074, 95 % CI: 1.696-2.537), diabetes duration (OR: 1.113, 95 % CI: 1.089-1.138), and hemoglobin (Hb)A1c (OR: 1.276, 95 % CI: 1.099-1.482) were independent DR risk factors. After excluding confounding factors and based on the β coefficient of the multivariate logistic analysis, the scale had a maximum score of 12 points. The area under the receiver operating characteristic curve was 0.753, the cut-off value was 4, the sensitivity was 66.5 %, and the coincidence rate was 70.9 %. To improve sensitivity, the cut-off value was lowered to 3; the sensitivity was 79.7 %, and the coincidence rate was 65.0 %.Conclusion: The simple scale had good sensitivity and the ability to identify DR patients.

There Is a Third Condition Aside from Survival and Death that Affects Outcome Statistics: Terminal Discharge

Background: Some patients discharged automatically are classified as terminal discharge, while their clinical outcome is survival, disrupting the results of clinical research. Methods: The data of this study were taken from inpatients admitted to the ICU of the First Medical Center of the People's Liberation Army General Hospital, Beijing, China from 2008-2017. We collected the data regarding medications used over the three days before discharge from the group of patients who survived and the group of patients who died, and the outcomes of all patients were recalculated by three classification algorithms (AdaBoosting, Pearson correlation coefficient, observed to expected ratio-weighted cosine similarity). Our basic assumption is that if the classification result is death but the actual in-hospital outcome is survival, the associated patient was likely terminally discharged. Results: The coincidence rate of the outcomes calculated by the AdaBoosting algorithm was 98.1%, the coincidence rate calculated by the Pearson correlation coefficient was 61.1%, and the coincidence rate calculated by the observed to expected ratio-weighted cosine similarity was 93.4%. When the three classification methods were combined, the accuracy reached 98.56%. Conclusion: The combination of clinical rules and classification methods has a synergistic effect on judgments of patients’ discharge outcomes, greatly saving time on manual retrieval and reducing the negative influence of statistics or rules.

Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis

Mycoplasma capricolumsubsp.subsp. capripneumonia (Mccp) and Mycoplasma mycoidessubsp.sbusp. capri (Mmc) cause caprine pleuropneumonia (CCPP) and mycoplasmal pneumonia in goats and sheep (MPGS), respectively. These diseases cannot be identified on clinical symptoms alone and it is laborious to distinguish them using biochemical methods. It is therefore important to establish a simple, rapid identification method for Mccp and Mmc. Here, we report a high-resolution melting (HRM) curve analysis using specific primers based on the Mmc 95010 strain MLC_0560 and Mccp F38 strain MCCPF38_00984 gene sequences. The method was highly specific with intra- and inter-batch coefficients of variation < 1%. The lower limit of detection for Mccp and Mmc was 55 copies/μL and 58 copies/μL, respectively. HRM and fluorescence qPCR results were compared using 106 nasal swabs and 47 lung tissue samples from goats (HRM-qPCR coincidence rate 94.8%; 145/153). Mycoplasma isolation and identification was performed on 30 lung tissue samples and 16 nasal swabs (HRM-culturing coincidence rate 87.0%; 40/46). HRM analysis was more sensitive than fluorescence qPCR and Mycoplasma isolation, indicating the practicality of HRM for accurate and rapid identification of Mccp and Mmc, and diagnosis and epidemiology of CCPP and MPGS.