Shuo Yang

Complete blood count reference intervals and age- and sex-related trends of North China Han population.

Abstract Background: Defining common reference intervals (RIs) are encouraging. The aim of this study is to establish RIs for complete blood count (CBC) in a Chinese Han population and probe their age- and sex-related CBC trends. Additionally, we will compare the CBC RIs of Han with those of other races. Methods: In total 1259 Han individuals (584 male and 675 female) were recruited in North China. CBC was processed on Sysmex XE-2100, Coulter LH750 and Mindray BC5800 whose traceability was well verified. The non-parametric 2.5th–97.5th percentiles RIs were calculated. Results: The RIs for CBC parameters did not show apparent analyzer-specificity, apart from mean cellular volume (MCV), mean platelet volume (MPV), plateletcrit (PCT) and platelet distribution width (PDW). Red blood cell (RBC), hemoglobin (HBG), hematocrit (HCT), mean cellular hemoglobin (MCH), and mean cellular hemoglobin concentration (MGHC) are higher in males; and their male mean values tend to drop after 40 years; conversely, the female mean values tend to rise. Platelet (PLT) is higher in females and tends to drop after 40 years in both sexes. White blood cell (WBC) and absolute count of neutrophils (NE) and monocytes (MO) are higher in males, but there is no apparent change with age. Lymphocytes (LY) absolute count declines with age in males, but the same change in females is not obvious. RIs for HBG and HCT are similar among Han, Nordic, US European and US Mexican populations and are lower in US Africans. WBC RIs for Han and US African populations are lower than that for US Europeans and US Mexicans. Conclusions: RIs for major blood cell parameters are not method-dependent; variations obviously exist in age, sex and race. Consequently, common RIs for most CBC parameters appear inapplicable.

Establishment of reference intervals of 24 chemistries in apparently healthy adult Han population of Northern China.

BACKGROUNDnThe availability of reference intervals is essential for physicians to interpret laboratory results. Most of our laboratory reference intervals are derived from data on the foreign population. We have studied the reference intervals of 24 common laboratory biochemical tests in an apparently healthy adult Han population of Northern China.nnnMETHODSn1364 healthy individuals between 20 and 79 years old were recruited. 24 different chemical analytes were tested by the Roche Modular P 800 analyzer. We stratified the populations by gender and age through applying exclusion criteria, and the reference intervals were obtained by statistical analysis.nnnRESULTSnReference intervals for 24 common analytes in a gender and age appropriate adult Han population of Northern China are reported.nnnCONCLUSIONnOur study provides adequate laboratory data on reference intervals. The results emphasize the significance of establishing population-based reference intervals for a clinical laboratory.

Protective Effects of Neutrophil Gelatinase–Associated Lipocalin on Hypoxia/Reoxygenation Injury of HK-2 Cells

BACKGROUNDnNeutrophil gelatinase-associated lipocalin (NAGL) was first extracted from neutrophil granules. Our previous study showed that the expression of NGAL mRNA and protein can be induced by hypoxia/reoxygenation. This study was designed to investigate the relationship between NGAL and hypoxia/reoxygenation injury pathologies in HK-2 cells.nnnMETHODSnThe effect of NGAL on the proliferation of HK-2 cell lines was analyzed with a MTT colorimetric assay. Cell-cycle distribution and measurement of the percentage of apoptotic cells were performed by flow cytometry after stained with propidium iodide and annexin V-fluorescein isothiocyanate. The expression of genes for apoptotic proteins Bcl-2, Bax, and caspase-3 was analyzed with real-time reverse-transcription polymerase chain reaction (RT-PCR). The expression of NGAL mRNA and protein was analyzed with real-time RT-PCR or Western blot, respectively.nnnRESULTSnHK-2 cells were treated with hypoxia/reoxygenation. HK-2 cells exhibited an increase in the number of cells in the G0/G1 phase and a decrease in the number of cells in the S and G2/M phases. The proliferation index is decreased. When HK-2 cells were treated with 200 ng/mL recombinant NGAL and hypoxia/reoxygenation, there were no effects on cell-cycle distribution. The ratio of early apoptotic cells in the control and hypoxia/reoxygenation groups were 1.1% and 26.5%, respectively. After the addition of 200 ng/mL recombinant NGAL, the ratio of early apoptotic cells in the hypoxia/reoxygenation group dropped to 19.6%. The expression of Bax/Bcl-2 ratio and caspase-3 were significantly higher in the hypoxia/reoxygenation compared with the control group. After the addition of 200 ng/mL recombinant NGAL, the levels of Bax/Bcl-2 ratio and caspase-3 decreased significantly compared with the control group.nnnCONCLUSIONSnThe action of NGAL against hypoxia/reoxygenation injury was due to inhibiting the apoptosis via inhibition of the expression of the genes of proapoptotic proteins Bax, Bcl-2, and caspase-3.

Autophagy activation attenuates renal ischemia-reperfusion injury in rats

Ischemia-reperfusion (I/R) injury is a leading cause of acute kidney injury (AKI), which is a common clinical complication but lacks effective therapies. This study investigated the role of autophagy in renal I/R injury and explored potential mechanisms in an established rat renal I/R injury model. Forty male Wistar rats were randomly divided into four groups: Sham, I/R, I/R pretreated with 3-methyladenine (3-MA, autophagy inhibitor), or I/R pretreated with rapamycin (autophagy activator). All rats were subjected to clamping of the left renal pedicle for 45u2009min after right nephrectomy, followed by 24u2009h of reperfusion. The Sham group underwent the surgical procedure without ischemia. 3-MA and rapamycin were injected 15u2009min before ischemia. Renal function was indicated by blood urea nitrogen and serum creatinine. Tissue samples from the kidneys were scored histopathologically. Autophagy was indicated by light chain 3 (LC3), Beclin-1, and p62 levels and the number of autophagic vacuoles. Apoptosis was evaluated by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method and expression of caspase-3. Autophagy was activated after renal I/R injury. Inhibition of autophagy by 3-MA before I/R aggravated renal injury, with worsened renal function, higher renal tissue injury scores, and more tubular apoptosis. In contrast, rapamycin pretreatment ameliorated renal injury, with improved renal function, lower renal tissue injury scores, and inhibited apoptosis based on fewer TUNEL-positive cells and lower caspase-3 expression. Our results demonstrate that autophagy could be activated during I/R injury and play a protective role in renal I/R injury. The mechanisms were involved in the regulation of several autophagy and apoptosis-related genes. Furthermore, autophagy activator may be a promising therapy for I/R injury and AKI in the future.

Trimester-specific coagulation and anticoagulation reference intervals for healthy pregnancy

BACKGROUNDnDue to the normal physiological need of pregnancy and childbirth, the haemostatic system of pregnant women is different from that of healthy non-pregnant women. The aim of this study was to establish trimester-specific reference intervals of coagulation screening tests and thrombophilia markers in pregnancies without complications of females with Han ethnicity from North China.nnnMETHODSnIn total 744 Han healthy pregnant women (first trimester 207 cases, second trimester 222 cases and third trimester 315 cases) and 121 healthy non-pregnant women were recruited in North China. Eight tests-activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen (Fib), d-Dimer, antithrombin (AT), protein C (PC) and free protein S (fPS)-were processed on ACL TOP automated coagulation analyzer. The non-parametric 2.5th-97.5th percentiles reference intervals were calculated to establish trimester-specific reference intervals.nnnRESULTSnThe reference intervals for APTT, PT, TT, Fib, d-Dimer, AT, PC, and fPS at first trimester were 26.4-41.9s, 9.7-12.5s, 11.7-17.0s, 2.38-4.44g/L, 0.01-0.31μg/mL, 72-120%, 29-150%, 21-143%, respectively. At second trimester, the reference intervals were 24.4-35.8s, 8.5-13.2s, 10.0-16.0s, 2.40-5.97g/L, 0.05-0.73μg/mL, 68-125%, 20-138%, 24-155%, respectively. At third trimester, the reference intervals were 25.6-34.9s, 8.6-12.4s, 11.1-15.5s, 2.79-5.91g/L, 0.14-2.82μg/mL, 56-119%, 20-134%, 17-140%, respectively. From the first trimester to the third trimester, APTT, PT and TT presented shortened trends, Fib and d-Dimer presented increasing trends, AT, PC and fPS activity presented decreasing trends, respectively.nnnCONCLUSIONSnThe trimester-specific reference intervals of coagulation screening tests and thrombophilia markers in pregnancies without complications of females with Han ethnicity from North China are presented in this study, which may provide effective evidence for doctors to accurately diagnose and treat the disease during pregnancy.

Establishment of reference intervals for complete blood count parameters during normal pregnancy in Beijing

To observe the changes of complete blood count (CBC) parameters during pregnancy and establish appropriate reference intervals for healthy pregnant women.

Neutrophil gelatinase-associated lipocalin worsens ischemia/reperfusion damage of kidney cells by autophagy.

Abstract This study aimed to explore the influence of neutrophil gelatinase-associated lipocalin on autophagy and its role in ischemia/reperfusion injury in human kidney-2 (HK-2) cells during acute kidney injury (AKI). HK-2 cells were given hypoxia/reoxygenation treatment for different times to simulate ischemia/reperfusion injury. Autophagy was evaluated by western blot and immunofluorescence of GFP-LC3. Cell viability was tested to reflect the degree of cell damage. The autophagy inhibitor 3-MA was used to inhibit autophagy and determine the role of autophagy in ischemia/reperfusion injury. HK-2 cells were hypoxia for 1u2009h, followed by reoxygenation treatment for 24u2009h. These cells were then exposed to human recombinant protein neutrophil gelatinase-associated lipocalin (NGAL) (50, 100, 200, 400, or 1000u2009ng/mL) with or without 3-MA. Our results showed that autophagy was induced by hypoxia treatment and was further enhanced by reoxygenation after hypoxia treatment. Cell viability was decreased with the inhibition of autophagy in the process. Autophagic flux was further induced with NGAL (>200u2009ng/mL), while cell viability declined in this condition. Cell viability was recovered when autophagy was inhibited. These results indicate that autophagy plays, in part, a protective role in renal ischemia/reperfusion injury. Furthermore, the data suggest that NGAL strengthens the level of autophagy in this process. Overall, a large quantity of NGAL produced by renal proximal tubular epithelial cells may induce excessive autophagy and increase renal ischemia/reperfusion injury in acute kidney injury.

Tissue Factor-bearing MPs and the risk of venous thrombosis in cancer patients: A meta-analysis.

Cancer patients with Tissue Factor (TF)-bearing MPs have been presented association with increased risk of venous thromboembolism (VTE), but results of these studies have not been consistent. We aimed to conduct a meta-analysis to assess the relationship between TF-bearing MPs and risk of VTE in patients with cancer. PubMed, Web of Science and EMBASE Databases were systematically retrieved up to1th June 2017. Two case-control studies and four cohort studies met the entry requirements in this analysis. The summary odd ratio (OR) were estimated by a random effect model. The overall OR was 1.76 (95% CI: 1.21–2.56, I2u2009=u200962.0%). The OR of case-control studies was 3.41 (95% CI: 1.45–8.02, I2u2009=u20090.0%) and that of cohort studies was1.53 (95% CI: 1.05–2.24, I2u2009=u200966.1%). The association between TF-bearing MPs and the risk of VTE in cancer patients was found in this meta-analysis. Publication bias testing and sensitivity subgroup analysis suggested that results of this meta-analysis were robustness. In conclusion, TF-bearing MPs were associated with increased risk of VTE in patients with cancer. Whereas, more well-designed studies and more comprehensive adjustments for confounders in further studies are warranted to affirm the association.

Age and Gender Tailored Cutoff Value of hs-cTnT Contributes to Rapidly Diagnose Acute Myocardial Infarction in Chest Pain Patients

BACKGROUNDnHs-cTnT concentrations are age and gender related, therefore establishment of age and gender optimized cutoff values for hs-cTnT might be essential. We aimed to define the age and gender specific hs-cTnT 99th percentile in reference population and assess its diagnostic and prognostic performance in patients with suspected AMI.nnnMETHODSnIn this prospective study, 1725 healthy individuals (18 - 97 years old) and 812 patients (15 - 98 years old) with chest pain with suspected AMI presenting to the emergency department were enrolled. The measurement of biomarkers was performed at presentation and at 3 - 4 hours according to clinical requirement. We stratified the reference population by age and gender through applying strict exclusion criteria. Clinical follow-up was obtained after 2 years.nnnRESULTSnThe 99th percentile of hs-cTnT according to age and gender in adult Han population of Northern China is reported. The cutoff values of hs-cTnT in for subjects < 70 years, 70 - 79 years, and ≥ 80 years was 16 ng/L, 38 ng/L, and 57 ng/L, respectively. Among the 812 patients with chest pain, according to the age and gender tailored cutoff value, the specificity and positive predictive value of AMI diagnosis were increased from 53.9% to 72.2% and 48.6% to 60.8%, compared to 14 ng/L commonly recommended. Cumulative 2-year survival rate for patients with hs-cTnT levels above 14 ng/L was 93.3% compared to 99.5% in patients below that level (p < 0.001). The same was observed for the age and gender tailored cutoff value which was 92.5% compared to 98.5%, respectively (p < 0.001).nnnCONCLUSIONSnAge and gender tailored cutoff value for hs-cTnT provides better diagnostic information, but yields no additional prognostic performance for risk prediction of death or major adverse cardiovascular events.

Angiotensin-Converting Enzyme Inhibitors' Influence on Antiplatelet Therapy of Clopidogrel in ACS.

BACKGROUNDnClopidogrel is a prodrug, the minority of which is converted to an active metabolite by hepatic cytochrome P450 (CYP2C19), however, most of it is metabolized to inactive substance by hepatic carboxylesterase1 (CES1). Meanwhile angiotensin-converting enzyme inhibitors (ACEIs) are mostly metabolized by CES1. We aimed to assess the impact of ACEIs on platelet inhibition by clopidogrel.nnnMETHODSnWe genotyped variants CES1, CYP2C19*2 and *3 in 502 patients with acute coronary syndrome (ACS) receiving clopidogrel therapy, and analyzed the effects of ACEIs on responsiveness to clopidogrel by the vasodilator-stimulated phosphoprotein (VASP) phosphorylation assay and ADP-stimulated impedance whole blood platelet aggregation assay.nnnRESULTSnIt showed that the allele frequency of CES1 c.428A was 0% in these patients. 45.22% (227/502) of these patients were carriers of CYP2C19*2 or CYP2C9*3 loss-of-function alleles. Among them, 57.71% (131/227) of the patients with CYP2C19 variants received ACEIs therapy. In a total of 502 patients, there was no difference in the VASP-PRI or the impedance whole blood platelet aggregation assay between the ACEIs group and non-ACEIs group [56.26 ± 14.55% versus 57.76 ± 13.56%, p = 0.241; 0 (0 - 2) Ω vs. 0 (0 - 2) Ω, p = 0.856]. In the CYP2C19 variant patients, there was no difference in the VASP-PRI or the impedance whole blood platelet aggregation assay between ACEIs group and non-ACEIs group [57.24 ± 15.12% versus 58.07 ± 13.90%, p = 0.667; 0 (0 - 2) Ω versus 0 (0 - 2) Ω, p = 0.536]. In the subgroups of ACS patients (unstable angina, non-ST-segment elevation myocardial infarction, ST-segment elevation myocardial infarction), there was no difference in the VASP-PRI between the ACEIs group and non-ACEIs group [55.81 ± 15.24% versus 58.37 ± 13.31%, p = 0.103; 55.76 ± 15.20% versus 49.09 ± 15.22%, p = 0.098; 58.13 ± 11.48% versus 61.87 ± 10.34%, p = 0.221], and there was no difference in the impedance whole blood platelet aggregation assay between ACEIs group and non-ACEIs group [0 (0 - 2) Ω versus 0 (0 - 2) Ω, p = 0.936; 0 (0 - 2) Ω versus 0 (0 - 2) Ω, p = 0.625; 0 (0 - 1.25) Ω versus 0 (0 - 1.5) Ω, p = 0.788].nnnCONCLUSIONSnIn our study, when ACEIs were used with clopidogrel, platelet response to clopidogrel was not affected. These findings suggest that the drug interaction between clopidogrel and ACEI is of little relevance in platelet function.