Establishing and evaluating an auto-verification system of thalassemia gene detection results.

The manual verification of gene tests is time-consuming and error prone. In this study, we try to explore a high-efficiency, clinically useful auto-verification system for gene detection of thalassemia. A series of verification elements were rooted in the auto-verification system. Consistency check was applied initially as one of the essential elements in our study. One hundred twenty-four archived cases were used to choose the consistency-check rules' indices from routine blood examination and hemoglobin electrophoresis by the receiver operating characteristic curves. Rule 1 and rule 2 established by the chosen indices were compared by their passing rate, consistency with manual validation, and error rate. Finally, 748 cases were used for verifying the system's feasibility by evaluating the passing rate, turn-around time (TAT), and error rate. The rule 2 had a higher passing rate (67.7% vs. 50.8%) and consistency (0.623 vs. 0.364) than the rule 1 with an error rate of zero. In a "live" valuation, the auto-verification system can reduce the TAT and error rate of verification by 51.5% and 0.13%, respectively, with a high passing rate of 82.8%. The auto-verification system for gene detection of thalassemia in this study can shorten the validation time, reduce errors, and enhance efficiency.

Residual Negative Pressure in Vacuum Tubes Might Increase the Risk of Spurious Hemolysis.

BACKGROUND: We planned a study to establish whether spurious hemolysis may occur when negative pressure remains in vacuum tubes. METHODS: Four tubes with different vacuum levels (-54, -65, -74, and -86 kPa) were used to examine blood drawn from one healthy volunteer; the tubes were allowed to stand for different times (1, 2, 3, and 4 hours). The plasma was separated and immediately tested for free hemoglobin (FHb). Thirty patients were enrolled in a verification experiment. RESULTS: The degree of hemolysis observed was greater when the remaining negative pressure was higher. Significant differences were recorded in the verification experiment. CONCLUSIONS: The results suggest that residual negative pressure might increase the risk of spurious hemolysis.

Molecular Epidemiological Characterization and Health Burden of Thalassemias in the Chaoshan Region, People's Republic of China.

Thalassemia is one of the most prevalent inherited disease in southern China. However, there have been only a few epidemiological studies of thalassemia in the Chaoshan region of Guangdong Province, People's Republic of China (PRC). A total of 6231 unrelated subjects in two main geographical cities of the Chaoshan region was analyzed for thalassemia. Seven hundred and thirty-six cases of suspected thalassemia carriers with microcytosis [mean corpuscular volume (MCV) <82.0 fL] were found by complete blood cell (CBC) count, and were tested by reverse dot-blot gene chip to reveal a total of 331 mutant chromosomes, including 278 α-thalassemia (α-thal) alleles and 53 ß-thalassemia (ß-thal) alleles. The most common α-thal mutations were the Southeast Asian (- -(SEA)), followed by the -α(3.7) (rightward) and -α(4.2) (leftward) deletions. The two most common ß-thal mutations were HBB: c.316-197C>T and HBB: c.126_129delCTTT, accounting for 69.81% of the ß-thal defects in the studied individuals. In addition, a rare mutation, Cap +1 (A>C) (HBB: c.-50A>C) was described for the first time in the Chaoshan region. Our results gave a heterozygote frequency of 5.31% for common α- and ß-thal in the Chaoshan region, and also indicated a higher prevalence of thalassemia with a heterozygote frequency of 6.29% in Chaozhou, followed by Shantou (3.37%). This study provided a detailed prevalence and molecular characterization of thalassemia in the Chaoshan region, and will be valuable for developing a strategy for prevention of thalassemia and reducing excessive health care costs in this area.

Assessment of Antiviral Properties of Peramivir against H7N9 Avian Influenza Virus in an Experimental Mouse Model.

The H7N9 influenza virus causes a severe form of disease in humans. Neuraminidase inhibitors, including oral oseltamivir and injectable peramivir, are the first choices of antiviral treatment for such cases; however, the clinical efficacy of these drugs is questionable. Animal experimental models are essential for understanding the viral replication kinetics under the selective pressure of antiviral agents. This study demonstrates the antiviral activity of peramivir in a mouse model of H7N9 avian influenza virus infection. The data show that repeated administration of peramivir at 30 mg/kg of body weight successfully eradicated the virus from the respiratory tract and extrapulmonary tissues during the acute response, prevented clinical signs of the disease, including neuropathy, and eventually protected mice against lethal H7N9 influenza virus infection. Early treatment with peramivir was found to be associated with better disease outcomes.

Genetic diversity of the 2013-14 human isolates of influenza H7N9 in China.

BACKGROUND: Influenza H7N9 has become an endemic pathogen in China where circulating virus is found extensively in wild birds and domestic poultry. Two epidemic waves of Human H7N9 infections have taken place in Eastern and South Central China during the years of 2013 and 2014. In this study, we report on the first four human cases of influenza H7N9 in Shantou, Guangdong province, which occurred during the second H7N9 wave, and the subsequent analysis of the viral isolates. METHODS: Viral genomes were subjected to multisegment amplification and sequenced in an Illumina MiSeq. Later, phylogenetic analyses of influenza H7N9 viruses were performed to establish the evolutionary context of the disease in humans. RESULTS: The sequences of the isolates from Shantou have closer evolutionary proximity to the predominant Eastern H7N9 cluster (similar to A/Shanghai/1/2013 (H7N9)) than to the Southern H7N9 cluster (similar to A/Guangdong/1/2013 (H7N9)). CONCLUSIONS: Two distinct phylogenetic groups of influenza H7N9 circulate currently in China and cause infections in humans as a consequence of cross-species spillover from the avian disease. The Eastern cluster, which includes the four isolates from Shantou, presents a wide geographic distribution and overlaps with the more restricted area of circulation of the Southern cluster. Continued monitoring of the avian disease is of critical importance to better understand and predict the epidemiological behaviour of the human cases.

Reduced risk of dyslipidaemia with oolong tea consumption: a population-based study in southern China.

Experimental studies have suggested that tea consumption could lower the risk of dyslipidaemia. However, epidemiological evidence is limited, especially in southern China, where oolong tea is the most widely consumed beverage. We conducted a population-based case-control study to evaluate the association between consumption of tea, especially oolong tea, and risk of dyslipidaemia in Shantou, southern China, from 2010 to 2011. Information on tea consumption, lifestyle characteristics and food consumption frequency of 1651 patients with newly diagnosed dyslipidaemia and 1390 controls was obtained using a semi-quantitative questionnaire. Anthropometric variables and serum biochemical indices were determined. Drinking more than 600 ml (2 paos) of green, oolong or black tea daily was found to be associated with the lowest odds of dyslipidaemia risk (P< 0.001) when compared with non-consumption, but only oolong tea consumption was found to be associated with low HDL-cholesterol levels. A dose-response relationship between duration of tea consumption and risk of dyslipidaemia (OR 0.10, 95% CI 0.06, 0.16), as well as that between amount of dried tea leaves brewed and risk of dyslipidaemia (OR 0.34, 95% CI 0.24, 0.48), was found. Moreover, consumption of oolong tea for the longest duration was found to be associated with 3.22, 11.99 and 6.69% lower blood total cholesterol, TAG and LDL-cholesterol levels, respectively. In conclusion, the present study indicates that long-term oolong tea consumption may be associated with a lower risk of dyslipidaemia in the population of Shantou in southern China.

Building and evaluating the autoverification of coagulation items in the laboratory information system.

BACKGROUND: The increasing number of specimens, the decreasing number of proficient staff, and clinical demands make autoverification important in the laboratory's future development. However, although autoverification has been widely applied, a full coverage of its technique is hard to achieve in a laboratory. The establishment of the autoverification rules and parameters is still unclear. Therefore, the aim of this study was to fill the vacancy by the method described below. METHODS: First, all logic processes and autoverification rules were established with reference to the Clinical and Laboratory Standards Institute (CLSI) document Auto10-A-Autoverification of Clinical Laboratory Test Results-Approved Guideline. Second, we established rules in the LIS, collected clinical specimens for validation, and assessed the results in a "live" environment. RESULTS: The whole passing rate of autoverification was 82.0% and the manual intervention rate was 18.0%. The highest passing rate of a single project was activated partial thromboplastin time (APTT), approximately 96.1%, followed by prothrombin time (PT), approximately 95.1%, and fibrinogen, approximately 90.9%. The turnaround time (TAT) was shortened by 31.8% (90 minutes vs. 132 minutes). CONCLUSIONS: Through implementing the autoverification, which accelerated verification and decreased the TAT and the odds of human review errors in the released results, we can save more time and concentrate on verifying the abnormal results and proceeding emergency tests.

Effectiveness of a standardized quality control management procedure for COVID-19 RT-PCR testing: a large-scale diagnostic accuracy study in China.

BACKGROUND: The study aimed to construct a standardized quality control management procedure (QCMP) and access its accuracy in the quality control of COVID-19 reverse transcriptase-polymerase chain reaction (RT-PCR). METHODS: Considering the initial RT-PCR results without applying QCMP as the gold standard, a large-scale diagnostic accuracy study including 4,385,925 participants at three COVID-19 RT-PCR testing sites in China, Foshan (as a pilot test), Guangzhou and Shenyang (as validation sites), was conducted from May 21, 2021, to December 15, 2022. RESULTS: In the pilot test, the RT-PCR with QCMP had a high accuracy of 99.18% with 100% specificity, 100% positive predictive value (PPV), and 99.17% negative predictive value (NPV). The rate of retesting was reduced from 1.98% to 1.16%. Its accuracy was then consistently validated in Guangzhou and Shenyang. CONCLUSIONS: The RT-PCR with QCMP showed excellent accuracy in identifying true negative COVID-19 and relieved the labor and time spent on retesting.

Glucose-6-phosphate dehydrogenase (G6PD) gene mutations detection by improved high-resolution DNA melting assay.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited disorder worldwide including southern China. G6PD gene mutations cause deficiency of the enzyme and a large spectrum of diseases. High-resolution DNA melting (HRM) assay was recently introduced as a rapid, inexpensive and effective method for genotyping. But there was a shortcoming of this method that hemizygous and homozygous genotypes were not easily distinguished from wild-types. Here we used improved HRM method for a small-scale screening of G6PD-deficient variants among people of Meizhou region. Then all amplicons were ascertained by direct DNA sequencing. These results indicated that HRM method was a major technical advance for G6PD mutations screening.

Diagnostic accuracy of autoverification and guidance system for COVID-19 RT-PCR results.

Background: To date, most countries worldwide have declared that the pandemic of COVID-19 is over, while the WHO has not officially ended the COVID-19 pandemic, and China still insists on the personalized dynamic COVID-free policy. Large-scale nucleic acid testing in Chinese communities and the manual interpretation for SARS-CoV-2 nucleic acid detection results pose a huge challenge for labour, quality and turnaround time (TAT) requirements. To solve this specific issue while increase the efficiency and accuracy of interpretation, we created an autoverification and guidance system (AGS) that can automatically interpret and report the COVID-19 reverse transcriptase-polymerase chain reaction (RT-PCR) results relaying on computer-based autoverification procedure and then validated its performance in real-world environments. This would be conductive to transmission risk prediction, COVID-19 prevention and control and timely medical treatment for positive patients in the context of the predictive, preventive and personalized medicine (PPPM). Methods: A diagnostic accuracy test was conducted with 380,693 participants from two COVID-19 test sites in China, the Hong Kong Hybribio Medical Laboratory (n = 266,035) and the mobile medical shelter at a Shanghai airport (n = 114,658). These participants underwent SARS-CoV-2 RT-PCR from March 28 to April 10, 2022. All RT-PCR results were interpreted by laboratorians and by using AGS simultaneously. Considering the manual interpretation as gold standard, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy were applied to evaluate the diagnostic value of the AGS on the interpretation of RT-PCR results. Results: Among the 266,035 samples in Hong Kong, there were 16,356 (6.15%) positive, 231,073 (86.86%) negative, 18,606 (6.99%) indefinite, 231,073 (86.86%, negative) no retest required and 34,962 (13.14%, positive and indefinite) retest required; the 114,658 samples in Shanghai consisted of 76 (0.07%) positive, 109,956 (95.90%) negative, 4626 (4.03%) indefinite, 109,956 (95.90%, negative) no retest required and 4702 (4.10%, positive and indefinite) retest required. Compared to the fashioned manual interpretation, the AGS is a procedure of high accuracy [99.96% (95%CI, 99.95-99.97%) in Hong Kong and 100% (95%CI, 100-100%) in Shanghai] with perfect sensitivity [99.98% (95%CI, 99.97-99.98%) in Hong Kong and 100% (95%CI, 100-100%) in Shanghai], specificity [99.87% (95%CI, 99.82-99.90%) in Hong Kong and 100% (95%CI, 99.92-100%) in Shanghai], PPV [99.98% (95%CI, 99.97-99.99%) in Hong Kong and 100% (95%CI, 99.99-100%) in Shanghai] and NPV [99.85% (95%CI, 99.80-99.88%) in Hong Kong and 100% (95%CI, 99.90-100%) in Shanghai]. The need for manual interpretation of total samples was dramatically reduced from 100% to 13.1% and the interpretation time fell from 53 h to 26 min in Hong Kong; while the manual interpretation of total samples was decreased from 100% to 4.1% and the interpretation time dropped from 20 h to 16 min at Shanghai. Conclusions: The AGS is a procedure of high accuracy and significantly relieves both labour and time from the challenge of large-scale screening of SARS-CoV-2 using RT-PCR. It should be recommended as a powerful screening, diagnostic and predictive system for SARS-CoV-2 to contribute timely the ending of the COVID-19 pandemic following the concept of PPPM.

Identification and Analysis of Potential Key Genes Associated With Hepatocellular Carcinoma Based on Integrated Bioinformatics Methods.

BACKGROUND: Hepatocellular carcinoma (HCC) is a type of primary liver tumor with poor prognosis and high mortality, and its molecular mechanism remains incompletely understood. This study aimed to use bioinformatics technology to identify differentially expressed genes (DEGs) in HCC pathogenesis, hoping to identify novel biomarkers or potential therapeutic targets for HCC research. METHODS: The bioinformatics analysis of our research mostly involved the following two datasets: Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). First, we screened DEGs based on the R packages (limma and edgeR). Using the DAVID database, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of DEGs were carried out. Next, the protein-protein interaction (PPI) network of the DEGs was built in the STRING database. Then, hub genes were screened through the cytoHubba plug-in, followed by verification using the GEPIA and Oncomine databases. We demonstrated differences in levels of the protein in hub genes using the Human Protein Atlas (HPA) database. Finally, the hub genes prognostic values were analyzed by the GEPIA database. Additionally, using the Comparative Toxicogenomics Database (CTD), we constructed the drug-gene interaction network. RESULTS: We ended up with 763 DEGs, including 247 upregulated and 516 downregulated DEGs, that were mainly enriched in the epoxygenase P450 pathway, oxidation-reduction process, and metabolism-related pathways. Through the constructed PPI network, it can be concluded that the P53 signaling pathway and the cell cycle are the most obvious in module analysis. From the PPI, we filtered out eight hub genes, and these genes were significantly upregulated in HCC samples, findings consistent with the expression validation results. Additionally, survival analysis showed that high level gene expression of CDC20, CDK1, MAD2L1, BUB1, BUB1B, CCNB1, and CCNA2 were connected with the poor overall survival of HCC patients. Toxicogenomics analysis showed that only topotecan, oxaliplatin, and azathioprine could reduce the gene expression levels of all seven hub genes. CONCLUSION: The present study screened out the key genes and pathways that were related to HCC pathogenesis, which could provide new insight for the future molecularly targeted therapy and prognosis evaluation of HCC.

Prognostic Model for Predicting Overall and Cancer-Specific Survival Among Patients With Cervical Squamous Cell Carcinoma: A SEER Based Study.

BACKGROUND: Cervical squamous cell carcinoma (CSCC) is the most common histological subtype of cervical cancer. The purpose of this study was to assess prognostic factors and establish personalized risk assessment nomograms to predict overall survival (OS) and cancer-specific survival (CSS) in CSCC patients. METHODS: CSCC patients diagnosed between 1988 and 2015 were identified in the Surveillance, Epidemiology, and End Results (SEER) database. Univariate and multivariate Cox proportional hazard regression models were applied to select meaningful independent predictors and construct predictive nomogram models for OS and CSS. The concordance index (C-index), calibration curve, and receiver operating characteristic (ROC) curve were used to determine the predictive accuracy and discriminability of the nomogram. RESULTS: A total cohort (n=17962) was randomly divided into a training cohort (n=11974) and a validation cohort (n=5988). Age, race, histologic grade, clinical stage, tumor size, chemotherapy and historic stage were assessed as common independent predictors of OS and CSS. The C-index value of the nomograms for predicting OS and CSS was 0.771 (95% confidence interval 0.762-0.780) and 0.786 (95% confidence interval 0.777-0.795), respectively. Calibration curves of the nomograms indicated satisfactory consistency between nomogram prediction and actual survival for both 3-year and 5-year OS and CSS. CONCLUSION: We constructed nomograms that could predict 3- and 5-year OS and CSS of CSCC patients. These nomograms showed good performance in prognostic prediction and can be used as an effective tool to evaluate the prognosis of CSCC patients, thus contributing to clinical decision making and individualized treatment planning.

Residual Negative Pressure in Vacuum Blood-Collection Tube and Hemolysis in Pediatric Blood Specimens.

OBJECTIVE: To determine a method to reduce specimen hemolysis rates in pediatric blood specimens. METHODS: A total of 290 blood specimens from pediatric patients were classified into the capped group or uncapped group. The hemolysis index and levels of lactate dehydrogenase (LDH) were measured using an automated biochemical analyzer. Also, we performed a paired test to measure the concentration of free hemoglobin in specimens from 25 randomly selected healthy adult volunteers, using a direct spectrophotometric technique. RESULTS: The hemolytic rate of capped specimens was 2-fold higher than that of uncapped specimens. We found significant differences for LDH. Also, there was a significant difference in the concentration of free hemoglobin in the random-volunteers test. CONCLUSIONS: Eliminating the residual negative pressure of vacuum blood-collection tubes was effective at reducing the macrohemolysis and/or microhemolysis rate.

High expression of forkhead box M1 (FOXM1) is a poor prognostic biomarker in lung adenocarcinoma.

BACKGROUND: Forkhead box M1 (FOXM1) is closely related to the formation and development of cancer. Because of differences in cellular origin, lung adenocarcinoma (ADC) and lung squamous cell carcinoma (SCC) usually exhibit different signatures. Therefore, it is essential to investigate the abnormalities of FOXM1 in the two subtypes separately. METHODS: Through the Oncomine and TCGA databases, we investigated the expression of FOXM1 mRNA, its prognostic value and possible mechanisms leading to its dysregulation. Furthermore, networks involving FOXM1 and its significantly altered neighboring genes were identified using the cBioPortal database. GO and KEGG enrichment analyses were performed using DAVID. RESULTS: Expression of FOXM1 mRNA was higher in lung tumor tissues than in normal tissues, and higher in SCC tissues than in ADC tissues. FOXM1 mRNA expression was correlated with N stage, TNM stage, age, sex and smoking history in ADC, but only correlated with N stage, age and sex in SCC. Survival analysis indicated that high expression of FOXM1 mRNA resulted to poor overall survival (OS) for ADC patients, but not for SCC patients. Cox regression analysis confirmed that FOXM1 mRNA expression was an independent prognostic indicator for ADC patients, and regression analysis identified a moderately positive correlation between FOXM1 mRNA levels and copy number alterations (CNAs), but a weakly negative association with DNA methylation. FOXM1 was mainly involved in cell cycle regulation, G2/M transition, G1/S transition and p53, PI3K-Akt and TGF-beta signaling pathway. CONCLUSIONS: High expression of FOXM1 mRNA might be an independent biomarker of poor OS in ADC patients.

Designing and evaluating autoverification rules for thyroid function profiles and sex hormone tests.

Purpose Following the analytical phase, the current practice of many hospital laboratories involves the manual verification of all test results followed by the production of the report. However, manual verification is a time-consuming and tedious process. In this paper, we provide a detailed description of how to design autoverification rules for thyroid function test profiles and sex hormones. Materials and methods We used DM2 (Data manager 2) to construct the algorithm and build the database for autoverification of thyroid function test profiles and sex hormones, with reference to Boolean logic, Auto 10-A and CLSI'88. The rules consist of checking quality control, instrument error flags, critical values, the analytical measurement range (AMR), the limit range, consistency check and delta check. Firstly, we established the rules in the DM2, collected clinical specimens for validation, then tested the rules in a 'live' environment. Results Agreement was achieved between manual verification by two senior laboratory personnel and verification using the autoverification rules in 99.78% of the cases. The total autoverification rate for all tests was 77.06%. Following implementation of the rules, the laboratory turnaround time (TAT) was reduced by 54.55% and staffing numbers fell from three to two whole time equivalents (WTE). Statistical analysis resulted in a kappa statistic of 0.99 ( P < 0.001). Moreover, after implementing the autoverification rules, the error rate fell to 0.04%, indicating that errors were almost completely eliminated. Conclusion Implementing autoverification rules can reduce TAT, minimize the number of samples that require manual verification and allow for a reduction in staffing numbers. It also allows laboratory staff to devote more time and effort to the handling of problematic test results and contributing to improved patient care.

Establishing and Evaluating Autoverification Rules with Intelligent Guidelines for Arterial Blood Gas Analysis in a Clinical Laboratory.

Arterial blood gas (ABG) analysis is important for acutely ill patients and should be performed by qualified laboratorians. The existing manual verifications are tedious, time-consuming, and prone to send wrong reports. Autoverification uses computer-based rules to verify clinical laboratory test results without manual review. To date, no data are available on the use of autoverification for ABG analysis. All autoverification rules were established according to AUTO10-A. Additionally, the rules were established using retrospective patient data, and then validated by actual clinical samples in a "live" environment before go-live. The average autoverification passing rate was 75.5%. The turnaround time (TAT) was reduced by 33.3% (27 min vs 18 min). Moreover, the error rate fell to 0.05% after implementation. Statistical analysis resulted in a kappa statistic of 0.92 ( p < 0.01), indicating close agreement between autoverification and senior technician verification, and the chi-square value was 22.4 ( p < 0.01), indicating that the autoverification error rate was lower than the manual verification error rate. Results showed that implementing autoverification rules with intelligent guidelines for ABG analysis of patients with critical illnesses could decrease the number of samples requiring manual verification, reduce TAT, and eliminate errors, allowing laboratorians to concentrate more time on abnormal samples, patient care, and collaboration with physicians.

miR-590 promotes the proliferation of HUMSCs and induces ECM synthesis by targeting Smad7.

MicroRNA (miR)-590 has been established to be a promoter of cell proliferation, migration and invasion, and an inhibitor of apoptosis in numerous cancer cell lines. However, its effects on non-cancer cells remain to be elucidated. miR-590 was transfected into human umbilical cord mesenchymal stem cells (HUMSCs), and the cell proliferation rate was determined using a Cell-Light 5-ethynyl-20-deoxyuridine Apollo 567 kit and the presence of extracellular matrix (ECM) proteins were detected using western blot analysis and immunofluorescence microscopy. Using bioinformatic analysis and dual-luciferase assays, the novel target miR-590 was identified. In addition, the effects of miR-590 on cell proliferation and ECM enhancement were also evaluated. The results of the present study demonstrated that miR-590 interacts directly with the 3'-untranslated region of Mothers Against Decapentaplegic Homolog 7 (Smad7), which is an important factor in transforming growth factor-ß signaling pathway. Overexpression of miR-590 downregulated Smad7 expression at the mRNA and protein level, and subsequently resulted in cell proliferation and ECM accumulation. Additionally, the transfection of small interfering RNA targeting Smad7 in HUMSCs produced similar effects on cell proliferation and ECM to the overexpression of miR-590. The results of the present study indicated that miR-590 affects HUMSC proliferation by directly targeting Smad7.

Development and Implementation of Autoverification Rules for ELISA Results of HBV Serological Markers.

Autoverification is a process of using computer-based rules to verify clinical laboratory test results without manual review. But to date, there are few published articles on the use of autoverification over the course of years in a clinical laboratory. In our study, we firstly described the development and implementation of autoverification rules for enzyme-linked immunosorbent assay (ELISA) results of hepatitis B virus (HBV) serological markers in a clinical immunology laboratory. We designed the autoverification rules for HBV by using Boolean logic on five clinically used serological markers in accordance with the framework of AUTO-10A, issued by the American Clinical Laboratory Standards Institute in 2006. The rules were written into the laboratory information system (LIS) and installed in the computer, so we could use the LIS to screen the test results. If the results passed the autoverification rules, they could be sent to doctors immediately. To evaluate the autoverification rules, we applied the real-time data of 11,585 patients with the autoverification rules. The autoverification rate of the five HBV serological markers was 79.5%. Furthermore, the turnaround time (TAT) was reduced by 38% (78 minutes vs. 126 minutes). The error rate was nearly eliminated. These results show that using LIS with autoverification rules can shorten TAT, enhance efficiency, and reduce manual review errors.

Effective expansion of forkhead box P3⁺ regulatory T cells via early secreted antigenic target 6 and antigen 85 complex B from Mycobacterium tuberculosis.

The expansion of CD4+ CD25+ forkhead box (FOX)P3+ regulatory T (Treg) cells has been observed in patients with Mycobacterium (M.) tuberculosis; however, the mechanism of expansion remains to be elucidated. The aim of the present study was to examine the role of the early secreted antigenic target 6(ESAT­6) and antigen 85 complex B (Ag85B) from M. tuberculosis on Treg cell expansion. To investigate the sensitivity of peripheral blood cultures to the M. tuberculosis ESAT­6 and Ag85B antigens, the proportion of circulating CD4+ CD25+ FOXP3+ Treg cells was determined using flow cytometry and the levels of FOXP3 mRNA were determined using reverse transcription quantitative polymerase chain reaction. The mRNA levels of FOXP3 and the proportion of circulating CD4+ CD25+ FOXP3+ Treg cells were increased in multiplicitous drug­resistant tuberculosis patients compared with those in healthy controls and patients with latent tuberculosis (TB) infection (LTBI) (P<0.001). The mycobacterial antigens ESAT­6 and Ag85B increased the expansion of the CD4+ CD25+ FOXP3+ Treg cells and the mRNA levels of FOXP3 in healthy controls and LTBI patients compared with the effect of Bacillus Calmette­Guerin (P<0.05). Additionally, the mRNA levels of FOXP3 were elevated in the LTBI patients following stimulations with the mycobacterial antigens (P=0.012). Therefore, the M. tuberculosis antigens ESAT­6 and Ag85B induced CD4+ CD25+ FOXP3+ Treg­cell expansion, particularly in patients with LTBI. These findings indicated that CD4+ CD25+ FOXP3+ Treg cells may have a primary role in the failure of the host immune system to eradicate M. tuberculosis.