Validation of a single specimen pneumatic tube system in the clinical laboratory.

The single-specimen pneumatic tube (PTS) is a commonly used rapid specimen delivery system in modern clinical laboratories. However, its impact on sample integrity and laboratory test results remains controversial. The installation and configuration of single-specimen PTS are unique to their institution. We sought to validate our single-specimen PTS by comparing routine chemistry, immunology, and hematology results with a repeat sample integrity index for manual transport. In 2023, 30 employees were randomly selected from the company medical examination, and three tubes of procoagulant serum samples and three tubes of EDTA anticoagulant blood samples were collected from each of them. Group A uses a single specimen PTS at 8 m/s, Group B uses a single specimen PTS at 15 m/s, and Group C uses manual transfer. Specimens from all three groups were simultaneously analysed for ALT, AST, TG, TC, LDL, K, NA, CI, TSH, hs-cTnT, NSE, Cyfra21-1 and haematological analysis. The differences between the three groups of NSE and Cyfra21-1 were statistically significant (P < 0.05). The differences of the rest of the items were not statistically significant. The difference in NSE was not statistically significant between groups A and B (P = 0.401), B and C, and C and A (P < 0.05). The difference in Cyfra21-1 was not statistically significant between groups A and B (P = 0.897), B and C (P = 0.052), and C and A (P = 0.145). Individual sample PTS should be validated for testing prior to use to ensure the results' accuracy.

Comparative study on the quality control effectiveness of AI-PBRTQC and traditional PBRTQC model in identifying quality risks.

Introduction: We compared the quality control efficiency of artificial intelligence-patient-based real-time quality control (AI-PBRTQC) and traditional PBRTQC in laboratories to create favorable conditions for the broader application of PBRTQC in clinical laboratories. Materials and methods: In the present study, the data of patients with total thyroxine (TT4), anti-Müllerian hormone (AMH), alanine aminotransferase (ALT), total cholesterol (TC), urea, and albumin (ALB) over five months were categorized into two groups: AI-PBRTQC group and traditional PBRTQC group. The Box-Cox transformation method estimated truncation ranges in the conventional PBRTQC group. In contrast, in the AI-PBRTQC group, the PBRTQC software platform intelligently selected the truncation ranges. We developed various validation models by incorporating different weighting factors, denoted as λ. Error detection, false positive rate, false negative rate, average number of the patient sample until error detection, and area under the curve were employed to evaluate the optimal PBRTQC model in this study. This study provides evidence of the effectiveness of AI-PBRTQC in identifying quality risks by analyzing quality risk cases. Results: The optimal parameter setting scheme for PBRTQC is TT4 (78-186), λ = 0.03; AMH (0.02-2.96), λ = 0.02; ALT (10-25), λ = 0.02; TC (2.84-5.87), λ = 0.02; urea (3.5-6.6), λ = 0.02; ALB (43-52), λ = 0.05. Conclusions: The AI-PBRTQC group was more efficient in identifying quality risks than the conventional PBRTQC. AI-PBRTQC can also effectively identify quality risks in a small number of samples. AI-PBRTQC can be used to determine quality risks in both biochemistry and immunology analytes. AI-PBRTQC identifies quality risks such as reagent calibration, onboard time, and brand changes.

Influential factors on urine EV DNA methylation detection and its diagnostic potential in prostate cancer.

The value of Extracellular vesicles (EVs) diagnostic markers is widely recognized. However, current research on EV DNA remains limited. This study investigates the biological properties, preprocessing factors, and diagnostic potential of EV DNA. We found that DNA positive vesicles account for 23.3% ± 6.7% of the urine total EV, with a large amount of DNA attached to the outside. EV DNA fragments are large, there is no significant effect on uEV DNA when store urine less than 6 h at 4°C. In addition, the influence of different EV extraction methods on methylation detection is also minor. More importantly, RASSF1A methylation in urine total EV DNA can distinguish between PCa and BPH, with an AUC of 0.874. Our results suggest the potential of urine EV DNA as a novel marker for PCa diagnosis. This provides a new idea for the study of urinary tumor markers.

Metabolic profiling in tissues and urine of patients with prostatic lesions and the diagnostic value of urine extracellular vesicles metabolites in prostate cancer.

BACKGROUND: Prostate cancer (PCa) lacks convenient and highly specific diagnostic markers. Although the value of extracellular vesicles (EV) in oncology is widely recognized, the diagnostic value of EV metabolites requires further exploration. This study aimed to explore the diagnostic value of urine EV (u-EV) metabolomics in PCa. METHODS: We first detected metabolites in paired tissues cells (cells), tissue EV (t-EVs), u-EVs, and urine samples in cohort 1 (8 PCa vs. 5 benign prostatic hypertrophy, BPH) to prob the feasibility of EV metabolites as diagnostic markers. We then analyzed the value of u-EVs as markers for PCa diagnosis and typing in the expanded sample cohort (60 PCa vs. 40 BPH). RESULTS: U-EV metabolites were more consistent with those in tissue-derived samples (cells and t-EVs) than those in urine, and more differential metabolites between BPH and PCa were identified in u-EV. Subsequently, we used a random forest model to construct a panel of six metabolites for PCa, which showed an area under the curve (AUC) of 0.833 in training cohort and 0.844 in validation cohort. We also found significantly differentially expressed metabolites between PCa subtypes (Gleason ≤ 7 vs. Gleason > 7 and localized vs. metastasis), demonstrating the value of EV metabolites in PCa typing and prognostic assessment. CONCLUSION: Metabolomic analysis of u-EVs is a promising source of noninvasive markers for PCa diagnosis.

Analysis between ABO blood group and clinical outcomes in COVID-19 patients and the potential mediating role of ACE2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become the most common coronavirus that causes large-scale infections worldwide. Currently, several studies have shown that the ABO blood group is associated with coronavirus disease 2019 (COVID-19) infection and some studies have also suggested that the infection of COVID-19 may be closely related to the interaction between angiotensin-converting enzyme 2 (ACE2) and blood group antigens. However, the relationship between blood type to clinical outcome in critically ill patients and the mechanism of action is still unclear. The current study aimed to examine the correlation between blood type distribution and SARS-CoV-2 infection, progression, and prognosis in patients with COVID-19 and the potential mediating role of ACE2. With 234 patients from 5 medical centers and two established cohorts, 137 for the mild cohort and 97 for the critically ill cohort, we found that the blood type A population was more sensitive to SARS-CoV-2, while the blood type distribution was not relevant to acute respiratory distress syndrome (ARDS), acute kidney injury (AKI), and mortality in COVID-19 patients. Further study showed that the serum ACE2 protein level of healthy people with type A was significantly higher than that of other blood groups, and type O was the lowest. The experimental results of spike protein binding to red blood cells also showed that the binding rate of people with type A was the highest, and that of people with type O was the lowest. Our finding indicated that blood type A may be the biological marker for susceptibility to SARS-CoV-2 infection and may be associated with potential mediating of ACE2, but irrelevant to the clinical outcomes including ARDS, AKI, and death. These findings can provide new ideas for clinical diagnosis, treatment, and prevention of COVID-19.

A reference interval study of ferritin and transferrin levels in donors from two blood centers.

Blood donors not only save the lives of patients but also play an important role in the development of medical and health services. Therefore, it is particularly important to pay attention to the blood health of blood donors who are at a high risk of iron deficiency. Detection of serum ferritin and transferrin is an important basis for the diagnosis of iron deficiency anemia. However, to the best of our knowledge, the levels of serum ferritin and transferrin, and the influencing factors, such as age and type of donation, in blood donors have not been clarified. In the present study, the serum ferritin and transferrin levels of donors from two blood centers were investigated. Demographic data were collected from the donors, and their serum ferritin and transferrin levels were tested. A total of 1,817 donors were enrolled and were eligible for evaluation. Reference intervals (RIs) for ferritin and transferrin were obtained from blood donors, and it was revealed that the ferritin and transferrin levels of blood donors were associated with age. Furthermore, serum transferrin levels were associated with the type of donation; the serum transferrin RI level was significantly higher in platelet-only donors compared with in whole blood donors. It was also demonstrated that ferritin levels were negatively associated with transferrin levels. The present study identified RIs for ferritin and transferrin levels in blood donors, and indicated that age and type of donation were important factors affecting ferritin and transferrin levels in blood donors. These findings may prove useful for blood donation recruitment and screening strategies in China, and could promote the health of blood donors.

Association between recipient survival and blood donor age after blood transfusion in a surgery intensive care unit: a multicenter randomized controlled trial study protocol.

BACKGROUND: Blood from younger individuals has been shown to improve physiological function in recipients in laboratory research, and many proteins from human peripheral blood show antisenescence capabilities. Thus, researchers have questioned whether blood from young donors is superior to blood from older donors. Blood transfusion is a key supportive therapy for trauma patients, and recent studies have reported the influence of blood donor age on recipient patient prognosis. Although some retrospective results found that blood from young donors improves survival, no influence of blood donor age was observed on outcomes in other study groups. The reasons for this discrepancy are complicated, but the fact that data were not obtained from randomized controlled trial (RCT) data should be considered. The current protocol and analysis method provide a feasible RCT design to evaluate the prognosis of severely ill surgery patients who were transfused with blood products from blood donors of different ages. METHODS: The current study is a pragmatic multicenter RCT (open, parallel-group, non-masked, superiority trial). Recruited surgery intensive care unit patients will be randomized into three groups and transfused with blood products from male donors of different ages (< 25, 25-45, and > 45 years). Survival time will be measured within 28 days. The survival characteristics, possible interaction between variables, and potential factors associated with death will be analyzed by Kaplan-Meier analysis, two-way ANOVA, and Cox proportional hazards model, respectively. TRIAL REGISTRATION: ChiCTR: ChiCTR190002. Registered on 22 March 2019. http://www.chictr.org.cn/showproj.aspx?proj=36867 .

A multicenter reference interval study of thromboelastography in the Chinese adult population.

BACKGROUND: Imprecise reference intervals (RIs) adversely impact the determination of the need for blood transfusion and clinical diagnosis and treatment of coagulopathy. However, there are few RI studies of thromboelastography (TEG) based on a standard protocol. The present multicenter study aimed to establish RIs for the adult Chinese population. METHODS: Healthy participants were recruited from 6 medical centers by non-probability sampling. Blood samples were subjected to laboratory TEG analysis. The Ichihara method, 2-level nested analysis of variance (ANOVA) (2N-ANOVA), and the latent abnormal values exclusion (LAVE) were used to define the RIs following recommendations of the Clinical and Laboratory Standards Institute and International Federation of Clinical Chemistry and Laboratory Medicine, Committee on Reference Intervals and Decision Limits. Multiple regression analysis was performed to explore sources of variation. RESULTS: A total of 507 healthy participants were enrolled into the study cohort. Twenty-five individuals with potential coagulopathy were secondarily excluded by LAVE. Smoking was related to reaction time, α angle, and coagulation index in the TEG test (P < 0.05). 2N-ANOVA revealed that the RIs of all 5 test items of TEG needed to be partitioned by age and sex. Finally, TEG RIs were derived both parametrically and nonparametrically for males or females and different age Groups, respectively. CONCLUSIONS: TEG RIs were established for the adult Chinese population using up-to-date methodology. The results will provide a useful and essential comparator for patients in the assessment of coagulation state, goal-directed blood transfusion therapy, and monitoring of the pharmacodynamic effects of anticoagulant drugs.

Performance of urinary neutrophil gelatinase-associated lipocalin, clusterin, and cystatin C in predicting diabetic kidney disease and diabetic microalbuminuria: a consecutive cohort study.

BACKGROUND: Tubular biomarkers have been regarded as emerging and promising markers for early diagnosis of diabetic kidney disease (DKD). The study was to determine the diagnostic capabilities of tubular biomarkers (urinary neutrophil gelatinase-associated lipocalin [NGAL], clusterin, and cystatin C) for DKD and diabetic microalbuminuria, and whether or not the tubular biomarkers appear earlier than microalbuminuria. METHODS: In this consecutive cohort study, 146 type 2 diabetes mellitus (T2DM) patients with a disease duration of ≥6 years were enrolled. Thirty age- and gender-matched subjects without any systemic diseases were recruited as the control group. Urinary samples collected before treatment were tested for NGAL, clusterin, and cystatin C. RESULTS: The levels of biomarkers were higher in patients with DKD (p < 0.001); and positively correlated with the urinary albumin creatinine ratio (UACR; p < 0.001). With respect to the diagnosis of DKD, the areas under the receiver operating characteristic curve (AUCs) for urinary NGAL, clusterin, and cystatin C were 0.816 (95% confidence interval [CI], 0.741-0.891), 0.775 (95% CI: 0.694-0.857), and 0.803 (95% CI: 0.722-0.884), respectively. The levels of urinary NGAL and cystatin C in the normoalbuminuria group (UACR <30 mg /g•Cr) were elevated compared with the control group, unlike urinary clusterin. There was no statistical difference in the levels of the three biomarkers between groups with different levels of haemoglobin A1C (HbA1c). The diagnostic AUCs for urinary NGAL, clusterin, and cystatin C in patients with diabetic microalbuminuria were 0.841 (95% CI: 0.775-0.907), 0.783(95% CI: 0.710-0.856), and 0.805 (95% CI: 0.733-0.877), respectively. CONCLUSIONS: Urinary NGAL, clusterin, and cystatin C may be promising biomarkers for diagnosing DKD and diabetic microalbuminuria. It is possible that urinary NGAL and cystatin C increase before the onset of microalbuminuria in T2DM patients.

Exosomal microRNA-141 is upregulated in the serum of prostate cancer patients.

PURPOSE: Novel biomarkers for the diagnosis of prostate cancer (PCa) are urgently required. Increasing evidence suggests that exosomal microRNAs (miRNAs or miRs) in serum may be potential noninvasive biomarkers for certain diseases. The objective of the present study was to investigate and assess whether exosomal miR-141 is an effective biomarker for human PCa. METHODS: In the present study, exosomes were isolated from the serum of patients with PCa, patients with benign prostate hyperplasia (BPH), and healthy volunteers. The total RNA was extracted from the exosomes and the level of miR-141 was analyzed by quantitative reverse transcription-polymerase chain reaction. The expression levels of miR-141 were compared between the whole serum and the serum exosomes of the three groups. Subsequently, the relevance of the exosomal expression of miR-141 to the clinicopathological factors in PCa was investigated. RESULTS: The expression of miR-141 was higher in exosomes compared with whole serum (control group, P=0.0003; BPH group, P=0.0016; PCa group, P<0.0001). The level of serum exosomal miR-141 was significantly higher in the patients with PCa compared with the patients with BPH and the healthy controls (3.85-fold, P=0.0007 and 4.06-fold, P=0.0005, respectively). In addition, the expression levels were significantly higher in metastatic PCa compared with localized PCa (P<0.0001). Receiver-operating characteristic curve revealed that the serum exosomal miR-141 yielded an area under the curve of 0.8694, with 80% sensitivity and 87.1% specificity in discriminating patients with metastatic PCa from the patients with localized PCa. CONCLUSION: Serum exosomes may serve as a more suitable material compared with the whole serum for measuring circulating miR-141 levels in patients with PCa. Exosomal miR-141 is upregulated in the serum from patients with PCa compared with patients with BPH or the healthy volunteers, and it may be a useful potential biomarker for the diagnosis of metastatic PCa.

Transcriptional and proteomic analyses of two-component response regulators in multidrug-resistant Mycobacterium tuberculosis.

Two-component systems (TCSs) have been reported to exhibit a sensing and responding role under drug stress that induces drug resistance in several bacterial species. However, the relationship between TCSs and multidrug resistance in Mycobacterium tuberculosis has not been comprehensively analysed to date. In this study, 90 M. tuberculosis clinical isolates were analysed using 15-loci mycobacterial interspersed repetitive unit (MIRU)-variable number tandem repeat (VNTR) typing and repetitive extragenic palindromic (rep)-PCR-based DNA fingerprinting. The results showed that all of the isolates were of the Beijing lineage, and strains with a drug-susceptible phenotype had not diverged into similar genotype clusters. Expression analysis of 13 response regulators of TCSs using real-time PCR and tandem mass spectrometry (MS/MS) proteomic analysis demonstrated that four response regulator genes (devR, mtrA, regX3 and Rv3143) were significantly upregulated in multidrug-resistant (MDR) strains compared with the laboratory strain H37Rv as well as drug-susceptible and isoniazid-monoresistant strains (P<0.05). DNA sequencing revealed that the promoter regions of devR, mtrA, regX3 and Rv3143 did not contain any mutations. Moreover, expression of the four genes could be induced by most of the four first-line antitubercular agents. In addition, either deletion or overexpression of devR in Mycobacterium bovis BCG did not alter its sensitivity to the four antitubercular drugs. This suggests that upregulation of devR, which is common in MDR-TB strains, might be induced by drug stress and hypoxic adaptation following the acquisition of multidrug resistance.

A C-terminal truncated mutation of licC attenuates the virulence of Streptococcus pneumoniae.

LicC has been identified as a virulence factor of Streptococcus pneumoniae. However, its role in virulence is still not fully understood because deletion of licC is lethal for the bacterium. In this study, a mutant with 78-bp truncation at the C-terminus of licC was obtained from a signature-tagged mutagenesis (STM) library. The mutant was viable with a large reduction in enzymatic activity as CTP:phosphocholine cytidylyltransferase detected in vitro using a firefly luciferase assay. The mutation attenuated the adhesion and invasion of S. pneumoniae ST556 (serotype 19F) to epithelial cells by 72% and 80%, respectively, and increased the phagocytosis by macrophages for 16.5%, compared to the parental strain. When the mutation was introduced into the encapsulated D39 strain (serotype 2), it led to attenuated virulence in mouse models either by intranasal colonization or by intraperitoneal infection. In addition, the phosphocholine (PCho) on cell surface was decreased, and the choline binding proteins (CBPs) were impaired, which may explain the attenuated virulence of the mutant. These observations indicate that C-terminus of licC is accounted for the main activity of LicC in PCho metabolism and is essential for the virulence of S. pneumoniae, which provides a novel target for drug design against pneumococcal infection.

Performance of urinary NGAL and L-FABP in predicting acute kidney injury and subsequent renal recovery: a cohort study based on major surgeries.

BACKGROUND: Acute kidney injury (AKI) is a frequent complication of major surgery. The current study evaluated the power of two biomarkers [urinary neutrophil gelatinase-associated lipocalin (NGAL) and liver-type fatty acid binding protein (L-FABP)] to detect the occurrence of AKI and to predict the recovery from renal dysfunction in a major surgery cohort. METHODS: In this prospective study, 199 patients undergoing major surgery were enrolled. Urinary samples collected from participants before surgery, and 0, 4, and 12 h and 1, 2, 7, and 14 days after surgery were tested for NGAL and L-FABP. RESULTS: Thirty-seven (18.6%) subjects developed AKI. Urinary NGAL and L-FABP were significantly increased from the time surgery was completed (p<0.05). The peak levels of NGAL and L-FABP occurred 12 and 4 h postoperatively (16.4- and 172.0-fold compared to baseline) in AKI group, respectively. The area under the receiver operating characteristic (ROC) curve (AUC) in NGAL (at 12 h), L-FABP (at 4 h), the most predictive model (NGAL at 12 h+L-FABP at 4 h), and the best combination at the same time point (12 h) was 0.83 [95% confidence interval (CI), 0.74-0.91], 0.85 (95% CI 0.77-0.93), 0.94 (95% CI 0.89-0.98), and 0.91 (95% CI 0.85-0.97), respectively. However, the largest AUC of single and combined biomarkers for predicting non-recovery after AKI only reached 0.70. CONCLUSIONS: Urinary NGAL and L-FABP can be used to detect AKI and combining NGAL and L-FABP may improve the diagnostic performance; however, NGAL and L-FABP may be poor predictors for renal recovery after AKI.

[Identification of LicC activity in pneumococcal infection].

AIM: To clone and express licC and its truncated form genes (25 amino acids from 3'-terminus were deleted and named deltaLicC) of Streptococcus pneumoniae, analyze the enzymatic activities of the proteins. METHODS: The recombinant plasmid pQE80-licC, pQE80-deltalicC were constructed, and the target proteins were expressed in E. coli BL21 under isopropy-beta-D-thiogalactoside (IPTG) induction. The proteins' activities were determined using bioluminescence test based on firefly luciferase assay system. RESULTS: The prokaryotic expression vector pQE80-licC, pQE80-deltalicC were successfully constructed and identified.The soluble proteins were obtained through inducing expression in E. coli BL21. It was showed that the activity of deltaLicC was markedly lower than that of LicC (P<0.05). CONCLUSION: The homemade bioluminescence assay method can miner the activity of LicC reliably and accurately. The important role of 25 amino acids from 3'-terminus for the activity of LicC was confirmed, and it was suggested that suppressing of LicC maybe was a useful method for treatment S.pneumococcal infection, especially drug resistant strain.