Improving the detection capability and efficiency of SARS-CoV-2 RNA specimens by the specimen turn-around process with multi-department cooperation.

Objective: Improving the detection capability and efficiency of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA specimens is very important for the prevention and control of the outbreak of Coronavirus disease 2019 (COVID-19). In this study, we evaluated the detection capability and efficiency of two outbreaks of COVID-19 before and after the process re-engineering in April and July 2022. Methods: This retrospective cross-sectional study involved 359,845 SARS-CoV-2 RNA specimens 2 weeks before and 2 weeks after the two outbreaks of COVID-19 in April and July. The number, transportation time and detection time of specimens, and the number of reports of more than 24 h were analyzed by SPSS software. Results: While 16.84% of people chose nasopharyngeal swabs (NPS) specimens, 83.16% chose oropharyngeal swabs (OPS) specimens to detect SARS-CoV-2 RNA. There were significant upward trends in the percentage of 10 sample pooling (P-10) from April before process re-engineering to July after process re-engineering (p < 0.001). Compared with April, the number of specimens in July increased significantly not only 2 weeks before but also 2 weeks after the outbreak of COVID-19, with an increase of 35.46 and 93.94%, respectively. After the process re-engineering, the number of reports more than 24 h in the 2 weeks before and after the outbreak of COVID-19 in July was significantly lower than that in April before process re-engineering (0% vs. 0.06% and 0 vs. 0.89%, both p < 0.001). Conclusion: The present study shows that strengthening the cooperation of multi-departments in process re-engineering, especially using the P-10 strategy and whole process informatization can improve the detection capability and efficiency of SARS-CoV-2 RNA specimens.

Diagnostic and prognostic value of blood inflammation and biochemical indicators for intrahepatic cholestasis of pregnancy in Chinese pregnant women.

Intrahepatic cholestasis of pregnancy (ICP) is a common liver disease during pregnancy, that has serious complications. This study aimed to compare the blood inflammation and biochemical markers of pregnant women with ICP in Southwest China and analyse their diagnostic value for ICP. A controlled cross-sectional study was conducted, and routine blood and biochemical indicators of 304 diagnosed ICP patients and 363 healthy pregnant women undergoing routine prenatal examination were assessed. The blood inflammatory indicators and biochemical indicators were compared between the ICP groups and normal groups. In this study, the levels of the ALT, AST, GGT, TBIL and DBIL biochemical indicators and the levels of WBC, neutrophils, NLR and PLR inflammatory indicators in the ICP group were significantly higher than those in healthy pregnant women (p < 0.001). The PA and lymphocytes of the ICP group were significantly lower than those of the normal group (p < 0.001). ROC curves showed that ALT and the NLR had higher predictive value for ICP. The GGT, TBA and NLR of pregnant women with ICP in the preterm group were significantly higher than those in the term group, and the combined NLR and TBA had a certain predictive value for preterm birth.

Molecular epidemiology of Streptococcus pneumoniae isosslated from children with community-acquired pneumonia under 5 years in Chengdu, China.

Streptococcus pneumoniae (S. pneumoniae) is one of the most common community-associated pathogens responsible for pneumonia in children. This retrospective study aimed to investigate the molecular characteristics of S. pneumoniae isolated from children with community-acquired pneumonia (CAP) under 5 years in Chengdu, China. Molecular characteristics of S. pneumoniae included serotype and virulence factor performed by using PCR method and sequence types (STs) determined by sequencing seven housekeeping genes. In addition, the potential relationships between molecular characteristics were depicted by minimum spanning tree and correspondence analysis. The prevailing serotypes were 19F (18.52%), 6B (17.59%), 19A (13.89%), 6A (6.48%) and 23F (5.56%) among 108 isolates. The overall coverage rates of 7-valent, 10-valent, 13-valent, 15-valent and 20-valent pneumococcal conjugate vaccines (PCVs) were 47.32, 48.1, 75, 75 and 78.7%, respectively. Meanwhile, the coverage rates of PCV13 among the isolates from CAP patients aged <1 year were high up to 84.2%. MLST analysis results showed that there were 56 different STs identified, of which the dominant STs were ST271 (22.22%) and ST320 (12.04%). Five international clones among STs were observed, including Spain23F-1, Spain6B-2, Taiwan19F-14, Netherlands3-31 and Denmark14-32. Additionally, most of the isolates carried ply, psaA, nanA, pavA, piaA and CC271 isolates expressed more of nanA than non-CC271 isolates. Moreover, there were strong relevant relationships among STs, serotypes and virulence factors. Considering serotypes and virulence factors together can be used as the foundation for the formulation of vaccine strategy.

Site-specific insertion of endonuclease recognition sites into amplicons to improve post-PCR analysis sensitivity of gene mutation.

Precise detection of low-frequency gene mutations surrounded by excess wild-type DNA is important in many aspects of medical fields. Most hybridization-based methods for high-resolution mutant allele analysis are hindered by competition of the complementary strand with single-strand probes for the target strand. Here, we demonstrate that site-specific insertion of endonuclease recognition sites into amplicons allows post-PCR generation of short dsDNA or ssDNA, whereby improves the sensitivity of both melting temperature analysis (MTA) and end-point detection following up. Using a three-staged PCR protocol, enrichment of target gene and incorporation of specific restriction sites in amplicons were ensued with hardly any loss in amplification efficiency and specificity. It enables simultaneous discrimination among a panel of totally 11 EGFR 19 exon deletion mutations via MTA after post-PCR digestion by either FokI only or cooperated with CRISPR-Cas12a, using SYBR green I. By replacement of one double-strand cleavage site with a nickase binding domain post-PCR generation of ssDNA of interest via strand displacement amplification (termed as iSDA) is realized. Our preliminary investigation shows that iSDA permits analysis of single nucleotide variants down to 0.1% allelic-frequency using end-point detection. Given the good compatibility with the majority of mutant-enrich PCR methods, we envision it would advance the current gene profiling technologies to a large extent.

Gestational diabetes mellitus is associated with blood inflammatory indicators in a Chinese pregnant women population.

OBJECTIVE: This study aimed to investigate relationships between blood inflammatory and metabolic biomarkers in pregnant Chinese women with gestational diabetes mellitus (GDM) in Southwest China and to compare the contributions of blood inflammatory indicators to GDM identification. SUBJECTS AND METHODS: In total, 1,347 pregnant women were divided by gestational week into early-, middle- and late-pregnancy groups. Fasting samples were collected for complete blood count (CBC) and biochemical indicator measurements. Receiver operating characteristic (ROC) curves were used to predict GDM. Multivariate logistic regression was used to examine the associations between blood inflammatory indicators and GDM prevalence in pregnant women. RESULTS: Significant differences were found in blood inflammatory indicators and metabolic markers in different pregnancy trimesters. For the pregnant women with GDM, FPG, 1 h-PG, 2 h-PG levels, NLR and PLR, WBC and neutrophil counts were statistically differences when compared with non-GDM groups. ROC curve analysis indicated that the NLR, PLR, WBC and neutrophil counts have predictive value for GDM from 24-28 gestational weeks. The FPG levels in 24th-28th weeks of pregnant women were positively correlated with the NLR, PLR, 1 h-PG, 2 h-PG levels and negatively correlated with lymphocyte count. Following adjustment for the age, PLR, and FPG level, multivariate logistic regression showed that the NLR was an independent factor predicting GDM in the 24th-28th weeks of pregnancy. CONCLUSION: The blood inflammatory indicators NLR, PLR, and WBC and neutrophil counts were higher in GDM women than in healthy pregnant women, suggesting that blood inflammatory indicators are associated with GDM.

Study on the Value of Blood Biomarkers NLR and PLR in the Clinical Diagnosis of Influenza a Virus Infection in Children.

BACKGROUND: The aim is to explore the value and significance of changes in neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) in the diagnosis and treatment of patients with influenza virus infection. METHODS: A total of 1,330 cases of influenza A diagnosed with the nucleic acid testing were collected according to the 2019 version of the influenza diagnosis and treatment regimen in our hospital from September 2020 to December 2020. During the same period, 1,330 cases of healthy subjects were used as controls. The colloidal gold method and fluorescent PCR were used to detect influenza A virus. The sysmex800i was used for routine blood test, and statistical analysis was then performed. RESULTS: Comparing the relevant indicators between the research group and the healthy control group showed that the differences in NLR, PLR, PLT, absolute lymphocyte values, etc. were all statistically significant (p < 0.001). Among them, the average results of NLR and PLR of the research group were all larger than those of the control group; the mean values of the absolute value of lymphocytes (x 109) and PLT (x 109) in the research group were all smaller than those of the control group. The NLR, PLR, LMR, age, and other parameters of the research group and the healthy control group were analyzed to determine whether there was influenza infection according to a binary logistic regression model. The results showed that the differences were not statistically significant except for age and LMR (p > 0.05) and did not enter the regression model. The differences in other parameters such as NLR and PLR were all statistically significant (all p < 0.001), which were all entered into the regression model. They were very significant for predictive diagnosis of influenza A. The areas under the ROC curve of NLR and PLR were 0.961 (95% CI: 0.953-0.968) and 0.749 (95% CI: 0.730-0.769), respectively; the sensitivity and specificity of NLR were 88.4% and 93.1%, respectively, and Youden's index was 0.815, the optimal diagnostic cutoff value was 1.478; the sensitivity and specificity of PLR were 56.70% and 89.60%, respectively; the Youden's index was 0.463, and the optimal diagnostic cutoff value was 124. CONCLUSIONS: NLR and PLR had a certain degree of accuracy in the diagnosis of viral infections in children with influenza A. The diagnostic effect of NLR was particularly good. In the early stage of the disease, cheap and easily available blood biomarkers can be used to diagnose influenza A. However, LMR had no diagnostic value for influenza A because the area under the curve was too small.

The Experience of the Fight Against COVID-19 in Clinical Laboratory Departments from Chengdu, China.

OBJECTIVE: In order to fight against coronavirus disease 2019 (COVID-19) better and to share our experience as a reference for clinical laboratory departments. METHODS: This was a retrospective study conducted in the clinical laboratory department of Chengdu Women's and Children's Central Hospital in Chengdu, China, from April 2020 to January 2021. The number of nucleic acid and antibody testing specimens of suspected COVID-19 cases was analyzed. The key points of suspected-case sample processing and detection in the clinical laboratory department were summarized. The laboratory was directly involved in the sample processing and testing of suspected cases, the release of reports, and the transfer of specimens to the fever clinic. RESULTS: The number of COVID-19 nucleic acid test specimens in our laboratory ranged from 102 to 2170 per day, and the number of antibody test specimens ranged from 24 to 391 per day. There were four main considerations in the treatment and detection of suspected-case specimens in the clinical laboratory: biosafety management in clinical laboratory departments, measures to ensure the health of the staff, the eight time points for processing suspected-case samples (turn-around time), and key points for the detection of suspected case specimens. CONCLUSION: The laboratory developed a protective process for COVID-19 antibody and nucleic acid detection during the pandemic. At present, the detection of COVID-19 antibodies and nucleic acids in the clinical laboratory department is orderly, and there have been no cases of laboratory infection.

Establishment of assay method- and trimester-specific reference intervals for thyroid hormones during pregnancy in Chengdu, China.

BACKGROUND: The reference intervals of thyroid hormone will change at different stages of pregnancy because of physiological alterations. On the other hand, the reference intervals of thyroid hormone will also change in different detection systems due to the manufacturer's methodology as well as a different race. The objective of this study was to establish the assay method- and trimester-specific reference intervals for thyroid-stimulating hormone, free thyroxine and free triiodothyronine for pregnant women in Chengdu. METHODS: A prospective, population-based cohort study involved 23,701 reference samples of pregnant women during the three trimesters and 8646 non-pregnant women with pre-pregnancy clinical and laboratory tests. The 2.5th and 97.5th percentiles were calculated as the reference intervals for thyroid-stimulating hormone, free thyroxine and free triiodothyronine at each trimester of pregnant women according to ATA Guidelines. RESULTS: The reference interval of thyroid-stimulating hormone in the 2.5th and 97.5th percentiles has a significant increasing trend from the first trimester, to second trimester and to third trimester, which was 0.08-3.79 mIU/L for the first trimester, and 0.12-3.95 mIU/L for the second trimester and 0.38-4.18 mIU/L for the third trimester, respectively (p < 0.001). However, the reference intervals of free thyroxine and free triiodothyronine in the 2.5th and 97.5th percentiles have significant decreasing trends from the first trimester, to second trimester and to third trimester, which were 11.87-18.83 pmol/L and 3.77-5.50 pmol/L for the first trimester, and 11.22-18.19 pmol/L and 3.60-5.41 pmol/L for the second trimester, and 10.19-17.42 pmol/L and 3.37-4.79 pmol/L for the third trimester, respectively (both p < 0.001). CONCLUSION: It is necessary to establish assay method- and trimester-specific reference intervals for thyroid-stimulating hormone, free thyroxine, and free triiodothyronine because the reference intervals of these thyroid hormones are significantly different at different stages of pregnancy.

Genetic investigation of haemoglobinopathies in a large cohort of asymptomatic individuals reveals a higher carrier rate for ß-thalassaemia in Sichuan Province (Southwestern China).

The incidence of haemoglobinopathy is high in China, especially south of the Yangtze River. However, the exact status of haemoglobinopathy in Sichuan is unknown. To carry out a detailed research of haemoglobinopathy in individuals living in Sichuan, 13,298 subjects without clinical symptoms who were living in Sichuan Province, with an age distribution of 5-73 years, were included in this study. Between March 2014 and July 2017, these subjects received examinations at the Medical Lab of Chengdu Women's & Children's Central Hospital. Mean corpuscular volume (MCV) < 82 fL or mean corpuscular haemoglobin (MCH) < 27 pg was used to indicate haemoglobinopathy carriers. Abnormal haemoglobin was screened by electrophoresis, and genes were sequenced to identify genotypes. Genotype diagnosis of alpha- and beta-thalassaemia was carried out by using PCR and shunt hybridization. There were 638 suspected haemoglobinopathy carriers (4.80%, 638/13,298). DNA sequencing identified 6 subjects with abnormal haemoglobin genotypes and 15 subjects with Hb E. The frequency of heterozygosity for thalassaemia was 4.12% (1.48% for α-thalassaemia and 2.61% for ß-thalassaemia) in Sichuan Province. The mutation spectrum of α-thalassaemia consisted of the five most common mutations: --SEA, -α3.7, -α4.2, αCS, and αQS. Seven types of ß-thalassaemia mutation were found in this study: CD41-42 (-TTCT) was the most frequent (28.47%), followed by 17 (A > T), -28 (A > G), and IVS-II-654 (C > T). The main abnormal haemoglobin genotype (HbE) and thalassaemia genotype (--SEA, CD41-42 (-TTCT)) were consistent with those in other regions of China, but the carrier rate of ß-thalassaemia in Sichuan was higher than that of α-thalassaemia.

Resource Recycling of Mn-Rich Sludge: Effective Separation of Impure Fe/Al and Recovery of High-Purity Hausmannite.

Groundwater treatment sludge is a Fe/Mn-rich waste generated in mass production in a groundwater treatment plant for potable water production. The conventional disposal of sludge, such as direct discharge into river/lake, sea, and landfill, is not environmentally sustainable. Herein, a novel method was proposed to effectively separate Fe/Al and recover Mn via a combined hydrochloric acid leaching and hydrothermal route. The sludge contained 14.6% Fe, 6.3% Mn, and 11.5% Al and was first dissolved in 5 M HCl to prepare a leaching solution. Second, the leaching solution was hydrothermally treated, in which 97.1% Fe and 94.8% Al were precipitated as hematite and boehmite and more than 98% Mn was kept. Increasing the reaction temperature to 270 °C was beneficial for Fe/Al removal. With the consumption of abundant H+, the reaction of added glucose and nitrate accelerated as the temperature increased. An optimal pH was utilized for Fe/Al hydrolysis and crystallization, leading to extensive removal of Fe/Al. Third, the residual solution was adjusted to pH 8.3 with NaOH, and approximately, 99.2% Mn was removed as hausmannite with a Mn content of 63.6%. This method exhibited efficient separation of impure Fe/Al from Mn-rich groundwater treatment plant iron mud, and the recycled high-purity hausmannite was a marketable active pharmaceutical ingredient.

Serum homocysteine and folate concentrations in early pregnancy and subsequent events of adverse pregnancy outcome: the Sichuan Homocysteine study.

BACKGROUND: Hyperhomocysteinemia may be a risk factor for endothelial dysfunction. Folate and vitamin B12 regulate the homocysteine metabolic process. This study aimed to evaluate the associations between subsequent events of adverse pregnancy outcome and early variables of homocysteine, folate, and vitamin B12 in pregnant women. METHODS: This multicenter, retrospective, case-control study involved 563 pregnant women with adverse pregnancy outcome and 600 controls. Adverse pregnancy outcomes included one or more of the following events: preeclampsia, preterm birth, low birth weight, and stillbirth. The associations between subsequent events of adverse pregnancy outcome and early variables of homocysteine, folate, and vitamin B12; metabolic parameters; inflammatory markers; anthropometrics; and lifestyle habits at 11-12 weeks of gestation were analyzed using the logistic regression model. RESULTS: Compared to the lower quartile homocysteine concentrations, the upper quartile homocysteine concentrations were associated with preeclampsia, preterm birth and low birth weight. On the contrary, the lower quartile folate concentrations were associated with preeclampsia, preterm birth and low birth weight compared with the upper quartile folate concentrations. The incidence of adverse pregnancy outcome increased progressively from the first to fourth homocysteine quartiles but decreased progressively from the first to fourth folate quartiles. After adjusting for confounding factors, multivariate logistic regression analysis showed that besides systolic blood pressure, diastolic blood pressure, body mass index and age, homocysteine (IV vs I quartile, aOR 5.89, 95% CI 4.08-8.51, P < 0.001), folate (IV vs I quartile, aOR 0.35, 95% CI 0.25-0.50, P < 0.001), folate supplementation (yes vs no, aOR 0.55, 95% CI 0.35-0.86, P = 0.010) during early pregnancy were independently associated with subsequent events of adverse pregnancy outcome, and vitamin B12 was rejected. Of these, the homocysteine revealed the highest odds ratio in all risk variables, and folate showed the lowest odds ratio in all protective variables. CONCLUSIONS: Higher homocysteine concentration and lower folate level during early pregnancy were associated with adverse pregnancy outcome. However, no association was found between vitamin B12 and adverse pregnancy outcome. Supplementation with folate in early pregnancy may reduce adverse pregnancy outcome.

Serum high concentrations of homocysteine and low levels of folic acid and vitamin B12 are significantly correlated with the categories of coronary artery diseases.

BACKGROUND: Homocysteine (Hcy) has been considered as an independent risk factor for coronary artery disease (CAD). Folic acid and vitamin B12 are two vital regulators in Hcy metabolic process. We evaluated the correlations between serum Hcy, folic acid and vitamin B12 with the categories of CAD. METHODS: Serum Hcy, folic acid and vitamin B12 from 292 CAD patients, including 73 acute myocardial infarction (AMI), 116 unstable angina pectoris (UAP), 103 stable angina pectoris (SAP), and 100 controls with chest pain patients were measured, and the data were analyzed by SPSS software. RESULTS: Compared to SAP patients, patients with AMI and UAP had higher Hcy levels with approximately average elevated (4-5) µmol/L, while SAP patients were approximately higher 8 µmol/L than controls. However, the levels of folic acid and vitamin B12 had opposite results, which in AMI group was the lowest, while in controls was the highest. CAD categories were positively correlated with Hcy (r = 0.286, p < 0.001), and negatively correlated with folic acid (r = -0.297, p < 0.001) and vitamin B12 (r = -0.208, p < 0.001). There were significant trend toward increase in the prevalence of high Hcy, low folic acid and vitamin B12 from controls, to SAP, to UAP, and to AMI. CONCLUSIONS: The present study provide the valuable evidence that high concentrations of Hcy and low levels of folic acid and vitamin B12 are significantly correlated with CAD categories.

Molecular Epidemiological Investigation of Thalassemia in the Chengdu Region, Sichuan Province, Southwest China.

Thalassemia is the most common inherited disease in southern China. However, this disorder is usually ignored by the health system in the Sichuan Province due to the lack of epidemiological data. To provide basic epidemiological data for thalassemia screening, genetic counseling, and prenatal diagnosis (PND) in the Chengdu region, a total of 3262 healthy subjects were assessed by complete blood count (CBC), reverse dot-blot gene chip, gap-polymerase chain reaction (gap-PCR), and PCR-DNA sequencing. A frequency of heterozygous thalassemia of 3.43% (112/3262) was found, of which 2.21% (72/3262) patients carried α-thalassemia (α-thal), 1.19% (39/3262) ß-thalassemia (ß-thal) and 0.3% (1/3262) hereditary persistence of fetal hemoglobin (Hb) (HPFH)/δß-thalassemia (δß-thal). Four types of α-thal mutations were found, the most prevalent being - -(SEA) (68.06%), followed by -α(3.7) (rightward deletion, 25.0%), Hb Quong Sze (Hb QS; HBA2: c.377 T > C) (4.17%), and -α(4.2) (leftward deletion, 2.78%). The seven ß-thal mutations included: codons 41/42 (-TTCT), HBB: c.126_129delCTTT (13/39, 33.33%); codon 17 (A > T), HBB: c.52 A > T (11/39, 28.95%); IVS-II-654 (C > T), HBB: c.316-197 C > T (9/39, 23.68%); -28 (A > G), HBB: c.-78 A > G (3/39, 7.69%); -29 (A > G), HBB: c.-79 A > G (1/39, 2.56%); codons 27/28 (+C), HBB: c.84_85insC (1/39, 2.56%), and the rare IVS-II-850 (G > T), HBB: c.316-1 G > T (1/39, 2.56%). Only one case of the Southeast Asian HPFH deletion was found. This is the first detailed molecular epidemiological survey of thalassemia in the Chengdu region, Sichuan Province, People's Republic of China (PRC).

Hyperhomocysteinemia as a metabolic disorder parameter is independently associated with the severity of coronary heart disease.

OBJECTIVES: To study the associations between hyperhomocysteinemia (HHcy) and the severity of coronary heart disease (CHD). METHODS: We retrospectively analyzed metabolic parameters, anthropometric variables, and life style habits in 292 CHD patients of different categories, and 100 controlled non-CHD patients with chest pain symptoms who were hospitalized in the Department of Cardiovascular Medicine, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, China between October 2013 and September 2014. RESULTS: The prevalence of HHcy in CHD patients was 79.1%, while only 5% of non-CHD patients had elevated serum homocysteine (Hcy) concentrations. The prevalence of HHcy significantly increased from 5% in non-CHD controls to 66% in the stable angina pectoris (SAP) group, to 81.9% in the unstable angina pectoris group, and to 93.15% in the acute myocardial infarction (AMI) group (p less than 0.001). After adjusting for confounding factors, multivariate logistic regression analysis showed that HHcy was independently associated with CHD category (AMI versus SAP, odds ratio [6.38], 95% confidence interval; 1.18-34.46). The Hcy was negatively correlated with folic acid (r=-0.67, p less than 0.001) and vitamin B12 (r=-0.56, p less than 0.001). Of the CHD patients with HHcy, 51.1% had low folic acid and 42% had low vitamin B12, 7 or 5 times higher than that of CHD patients with normal-low Hcy concentrations (p less than 0.001). CONCLUSION: Hyperhomocysteinemia is independently associated with the severity of CHD, and significantly correlated with low status of folic acid and vitamin B12 in CHD patients.

Elevated serum myeloperoxidase activities are significantly associated with the prevalence of ACS and High LDL-C levels in CHD patients.

AIM: Recent researches have shown that myeloperoxidase (MPO) is a potential inflammatory risk factor for coronary heart disease (CHD). In the present study, the possible associations of MPO with acute coronary syndrome (ACS), low density lipoprotein cholesterol (LDL-C) and other risk factors in CHD patients were investigated. METHODS: Five hundred thirty-six CHD patients [363 ACS and 173 stable angina pectoris (SAP)] and 181 non-CHD patients confirmed by coronary angiography were enrolled in this study. The association study was performed by logistic regression analysis. RESULTS: ACS patients had significantly higher MPO activities than SAP and non-CHD patients (p < 0.001). The area under the receiver operating characteristic (ROC) curve of MPO for diagnosing ACS was 0.84 (95% CI: 0.80-0.88, p < 0.001). The optimal cut-off value (sensitivity; specificity) of MPO was 164.77 ng/mL (79.1% and 82.1%). LDL-C (III versus I tertile, OR: 3.24, 95% CI: 1.67-6.29, p = 0.001) and ACS (yes versus no, OR 2.74, 95% CI: 1.71-4.39, p < 0.001) were significantly associated with elevated serum MPO activities, which had the highest odds ratio in quantitative and qualitative variables, respectively. There were significant increase trends in the prevalence of ACS and high LDL-C levels from I to III MPO tertile, which were 46.4% and 8.9% for I, 68.0% and 31.5% for II, 88.8% and 59.8% for III tertile, respectively (p < 0.001). CONCLUSION: The present study provides epidemiological evidence that elevated serum MPO activities are significantly associated with the prevalence of ACS and high LDL-C levels in CHD patients, and MPO may be a potential early warning marker for ACS.

Ultrasensitive electrochemical immunosensor for HE4 based on rolling circle amplification.

A novel electrochemical immunoassay system for the detection of human epididymis-specific protein 4 (HE4) was developed. A chitosan-titanium carbide (TiC) nanocomposition film was first electrodeposited onto a tin-doped indium oxide (ITO) electrode at a constant potential. Gold (Au) nanoparticles were then electrodeposited on the surface of the chitosan-TiC film by cyclic voltammetry (CV). The capture antibody (anti-HE4) was adsorbed onto the Au and TiC nanoparticles. After a specific sandwich immunoreaction among the capture antibody, HE4, and biotinylated secondary antibody, biotinylated primer DNA was immobilized on the secondary antibody by biotin-streptavidin system. Appropriate amounts of circular template DNA and biotinylated primer DNA were used for rolling circle amplification (RCA) under optimal conditions. The RCA products provided a large number of sites to link DNA detection probes. Doxorubicin hydrochloride intercalated the CG-GC steps between the RCA products and the DNA detection probes, which was monitored by differential pulse voltammetry (DPV) based on the current signal of doxorubicin hydrochloride. With the above-mentioned amplification factors, the current responded to HE4 linearly in the concentration range of 3-300 pM under optimal detection conditions, with a detection limit of 0.06 pM. Stepwise changes in the microscopic features of the surfaces and electrochemical properties upon the formation of each layer were confirmed by scanning electron microscopy (SEM), atomic force microscopy (AFM), and electrochemical impedance spectroscopy (EIS). This system was successfully employed for the detection of HE4 with good accuracy and renewable ability.

Blockade of NKG2D synergized with CTLA4-Ig in promoting long-term graft survival in murine models of cardiac transplantation.

BACKGROUND: Blockade of B7-CD28 interaction with CTLA4-Ig could prolong allograft survival in cardiac transplantation. NKG2D is an activating or coactivating receptor on NK cells, γδ T, and CD8 T cells and played an important role in transplant immunity. METHODS: C57BL/6 (wild type and γδ or interleukin (IL)-17) mice were transplanted with allogeneic BALB/c hearts and treated with CTLA4-Ig alone or in combination with anti-NKG2D monoclonal antibodies. The survival of grafts was monitored daily by abdominal palpation until the complete cessation of cardiac contractility. RESULTS: We found that wild type recipient treated with anti-NKG2D monoclonal antibodies plus CTLA4-Ig showed significantly prolonged survival cardiac allograft (>90 days, P<0.001). These in vivo results in combined therapy group correlated with low expression of interferon-γ whereas increased expression of IL-4 and alternatively activated macrophage markers. Furthermore, with blockade of NKG2D, the number of IL-17-producing γδ T cells was significantly reduced, which was demonstrated as the main source of IL-17 production. And in our γδ and IL-17 murine cardiac transplantation models, we found that γδ or IL-17 deficiency could significantly prolong cardiac allograft survival. CONCLUSION: Blockade of NKG2D is effective in synergizing with CTLA4-Ig to promote long-term cardiac allograft survival in mice, and this effect is associated with decreased infiltration of IL-17-producing γδ T cells.