Objective To evaluate the limit of blank (LoB),limit of detection (LoD),limit of quantitation(LoQ)and functional sensitivity (FS)of prealbumin (PA)detected by the Roche Modular P automatic biochemical analyzer.Methods According to the EP17A file of the American Clinical and Laboratory Standards Institute (CLSI),saline as the blank sample and a series of low con-centration samples were detected by the Roche Modular P automatic biochemical analyzer for determining LoB,LoD and LoQ.And FS was determined based on the domestic universal method .Results LoB of PA was 16.35 mg/L,LoD was 18.23 mg/L,LoQ was temporarily unable to evaluate and FS was 25.00 mg/L.The report scope and the report mode in clinic were affirmed by combi-ning with the low value of the reportable scope.Conclusion LoD of PA detected by the Roche Modular P automatic biochemical an-alyzer is established,which provides more valuable information for clinical diagnosis and treatment.Conducting the comparison of different evaluation methods determines the advantages and limitation of the practical application of different methods.

BACKGROUND: Ischemia-reperfusion injury in the myocardium after cardiac arrest (CA) and cardiopulmonary resuscitation (CPR) is an important pathologic basis of post-cardiac arrest of syndrome (PCAS), and apoptosis is one of the major mechanisms in myocardial ischemia-reperfusion injury. To lessen myocardial ischemia-reperfusion injury after cardiac arrest and CPR, it is important to reduce energy consumption and to increase energy supply in the myocardium. This study aimed to observe changes of cell apoptosis and expression of Bcl-2 and Bax protein on the myocardium after CPR in rats, and the protective effects of different doses of exogenous phosphocreatine (creatine phosphate, CP) on them. METHODS: A total of 32 male adult Sprague-Dawley rats were randomly divided into 4 groups: control group (group A), CPR group (group B), low-dose CP group (group C, CP 0.5 g/kg at beginning of CPR and 1.0 g/kg at 2 hours after CPR) and high-dose CP group (group D, CP 1.0 g/kg at beginning of CPR and 2.0 g/kg at 2 hours after CPR). Cardiac arrest was induced by asphyxiation and CPR started at 7 minutes after asphyxiation in groups B, C and D. Myocardium samples were taken at 24 hours after CPR. Cardiomycytic apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method. The expression of Bcl-2 and Bax protein was measured by immunohistochemistry. RESULTS: Cardiomyocytic apoptosis index (AI) and expression of Bcl-2 and Bax protein increased more significantly in groups B, C and D than in group A (P<0.01), but Bcl-2/Baxratio significantly decreased (P<0.01). Cardiomyocytic AI and expression of Bcl-2 and Bax protein decreased more significantly in groups C and D than in group B (P<0.01), but Bcl-2/Bax ratio increased more significantly (P<0.01). Cardiomyocytic AI and expression of Bcl-2 and Bax protein decreased more significantly in group D than in group C (P<0.05), but Bcl-2/Bax ratio increased more significantly (P<0.05). CONCLUSION: Exogenous phosphocreatine, especially at a large dose, could inhibit cardiomyocytic apoptosis and alleviate myocardial injury after CPR in rats.

Humans ; Liver Transplantation ; Transplantation, Autologous

AIM:To investigate the role of hydrogen molecule on apoptosis-related proteins in glomerular me-sangial cells cultured with high glucose and to explore its possible mechanism.METHODS:Mouse glomerular mesangial cells cultured in vitro were divided into 4 groups:normal control group(C group,5.5 mmol/L glucose),mannitol group(G group,5.5 mmol/L glucose+19.5 mmol/L mannitol),high glucose group(H group,25 mmol/L glucose),high glu-cose+hydrogen-rich water group(HH group,25 mmol/L glucose+hydrogen-rich water),and cultured for 48 h.The pro-tein levels of Bax,Bcl-2,cleaved caspase-3,nuclear factor E2-related factor-2(Nrf2),heme oxygenase-1(HO-1)and NAD(P)H:quinone oxidoreductase-1(NQO-1)were determined by Western blot ,and the mRNA expression of HO-1 and NQO-1 was determined by RT-PCR.The level of intracellular reactive oxygen species(ROS)was detected by dihydro-ethidium method,and the activity of superoxide dismutase(SOD)was measured by WST-8 assay.RESULTS:Compared with C group,the protein levels of Bax and cleaved caspase-3 were up-regulated,and Bcl-2 was down-regulated in H group(P <0.05).No significantly difference of the protein levels mentioned above between C and HH group was observed. Compared with H group,the protein levels of Bax and cleaved caspase-3 were down-regulated,and Bcl-2 was up-regulated in HH group(P <0.05).The level of intracellular ROS was higher and the activity of SOD was lower in H group than those in C group(P<0.05).However,there was no difference of the SOD activity between C group and HH group.The level of intracellular ROS decreased and the activity of SOD increased in HH group as compared with H group(P<0.05). Compared with C group,clearly reduced protein expression of Nrf2,HO-1 and NQO-1,and decreased mRNA expression of HO-1 and NQO-1 in H group were observed(P<0.05).Compared with H group,the protein levels of Nrf2,HO-1 and NQO-1 as well as the mRNA levels of HO-1 and NQO-1 were obviously increased in HH group(P<0.05 ).CONCLU-SION:Hydrogen molecule inhibits the expression of pro-apoptotic proteins and induces the expression of anti-apoptotic pro-teins in glomerular mesangial cells cultured with high glucose.The mechanism may be related to activation of Nrf 2 signaling pathway.

This study was purposed to investigate the clinical significance of the amount and activated status of T cell subsets, B cells, NK cells in peripheral blood from patients with myelodysplastic syndrome (MDS). The proportion of T cells, B cells, NK cells in peripheral blood from 30 patients with MDS and their surface activation markers of CD28, CD45RA, CD45RO, CD69, HLA-DR were analyzed by flow cytometry. Twenty-two patients were in the low risk group (RA + RAS) while eight patients were in the high risk group (RAEB + RAEBT). The result showed that the amounts of T cells (CD3+ cells) in peripheral blood from patients with MDS were lower than those in control group. The amounts of naive CD4+ cells (CD4+ CD45RA+ cells) in MDS patients were lower than those in control. The expression rates of early activation marker (CD69) and late activation marker (HLA-DR) on CD3+ cells in MDS patients were significantly higher than those in control. The abnormalities of the immunologically competent cells were mainly observed in the low risk group (RA + RAS), and were characterized by the high expression rates of CD69+ and HLA-DR+ on CD3+ cells, the decrease of B cell amounts. The amount abnormalities of T cell subsets were mainly observed in high risk group (RAEB + RAEBT), and were characterized by the decrease of CD3+ cells and CD3+ CD4+ CD8- cells (Th cells) amounts without high expression of the CD69 and HLA-DR, the decrease of NK cells amounts. It is concluded that there are the abnormalities of T cell subsets and function in the patients with MDS and may change with disease progression, so the measurement of amount and activated status of T cell subsets in peripheral blood from MDS patients can have predictive role for diagnosis of disease progression and guide of therapy.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; immunology ; Antigens, Differentiation, T-Lymphocyte ; immunology ; B-Lymphocytes ; immunology ; CD3 Complex ; immunology ; Female ; HLA-DR Antigens ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Lectins, C-Type ; Lymphocyte Activation ; immunology ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; T-Lymphocyte Subsets ; immunology

OBJECTIVETo evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSChimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR).

RESULTSOn day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred.

CONCLUSIONSerial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

Adolescent ; Adult ; Child ; Chimerism ; Female ; Graft Rejection ; immunology ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Male ; Middle Aged ; Recurrence ; T-Lymphocytes ; immunology ; Young Adult

OBJECTIVETo evaluate the association of C-reactive protein/albumin ratio (CAR) with the prognosis of patients with colorectal cancer and compare the prognostic value of CAR with other inflammation-based prognostic scoring systems.

METHODSWe retrospectively evaluated 163 newly diagnosed colorectal cancer patients in Nanfang Hospital between January, 2007 and December, 2014. All recommended cutoff values of the clinicopathological factors were defined using receiver- operating characteristic (ROC) curve analyses. We evaluated the prognostic value of CAR in comparison with Glasgow Prognostic Score (GPS) and neutrophil lymphocyte ratio (NLR) with the area under the ROC curve. Univariate and multivariate analyses using the Cox proportional hazards model were performed to identify the factors closely associated with overall survival of the patients. Kaplan-Meier analysis was used to compare overall survival curves between patients with a high CAR and those with a low CAR.

RESULTSThe recommended cutoff value of CAR was 0.132. Kaplan-Meier analysis and log rank test demonstrated a significant difference in the overall survival between patients with a low CAR (<0.132) and those with a high CAR (≥0.132) (2157.0∓395.3 vs 1661.0∓136.4 days, P<0.001). The area under the ROC curve of CAR, NLR and GPS was 0.656, 0.550 and 0.642, respectively, indicating a better prognostic value of CAR. Univariate analyses showed that age, C-reactive protein, albumin, CAR, NLR, GPS, platelet, TMN stage, Dukes stage and chemotherapy regimens were associated with the overall survival of the patients (P<0.05). Multivariate analyses showed that TMN stage [HR=1.689 (95%CI: 1.146-2.488), P=0.008] and Dukes stage [HR=2.447 (95%CI: 1.349-4.441), P=0.003] were associated with the overall survival of the patients.

CONCLUSIONSSimilar to the previously reported inflammation-based prognostic systems (GPS and NLR), CAR is useful for predicting the survival of patients with colorectal cancer and can be complementary to the two prognostic scoring systems.

OBJECTIVETo analyze related factors which affect GPA mutation frequency of workers exposed to benzene, with the Glycophorin A (GPA) mutation assay and explore the possibility of GPA mutation frequency as an index of predicting the risk of benzene poisoning.

METHODSThe erythrocytes were bound with fluorescent-labeled monoclonal antibody after isolated and fixed from the peripheral blood, and then the GPA mutation assay was performed using the flow cytometry (FCM). The related factors of GPA mutation frequency were analyzed by statistical methods.

RESULTSThe GPA mutation frequency of chronic benzene poisonings was significantly higher than that of their controls (P < 0.05). Significant direct correlation was found between age, length of service, accumulative exposure score and the GPA mutation frequency of workers exposed to benzene (P < 0.01). However, there was significantly inverse correlation between the 3AB index and the GPA mutation frequency (GPAN0: r(s) = -0.589, P < 0.01, GPANN: r(s) = -0.615, P < 0.01). In the multiple factor regression analysis on GPA mutation frequency, benzene exposure and individual susceptibility both entered model of multiple factors analysis, the coefficient of determination of benzene-exposed workers was 0.819.

CONCLUSIONExposure to benzene and individual susceptibility are the most important factors that affect GPA mutation frequency. GPA mutation frequency increases with the benzene exposure and individual susceptibility.

Benzene ; poisoning ; Glycophorin ; genetics ; Humans ; Mutation ; Mutation Rate ; Occupational Exposure

OBJECTIVETo investigate the effect of ceftriaxone on the intestinal epithelium and microbiota in mice in the early-life stage, as well as the recovery of the intestinal epithelium and reconstruction of intestinal microbiota in adult mice.

METHODSA total of 36 BALB/C neonatal mice were randomly divided into control group and experimental group, with 18 mice in each group. The mice in the experimental group were given ceftriaxone 100 mg/kg every day by gavage within 21 days after birth. Those in the control group were given an equal volume of normal saline by gavage. Immunohistochemistry was used to measure the expression of Ki67, Muc2, and ZO-1 in the intestinal epithelium. qPCR and next-generation sequencing were used to analyze the overall concentration and composition of fecal bacteria.

RESULTSAfter 21 days of ceftriaxone intervention, the experimental group had a significant reduction in body weight, a significant reduction in the expression of Ki67 and ZO-1 and a significant increase in the expression of Muc2 in intestinal epithelial cells, a significant reduction in the overall concentration of fecal bacteria, and a significant increase in the diversity of fecal bacteria compared with the control group (P<0.05). Firmicutes was the most common type of fecal bacteria in the experimental group, and there were large amounts of Staphylococcus and Enterococcus. The experimental group had a certain degree of recovery of the intestinal epithelium, but there were still significant differences in body weight and the structure of intestinal microbiota between the two groups at 56 days after birth (P<0.05).

CONCLUSIONSEarly ceftriaxone intervention significantly affects the development of the intestinal epithelium and the construction of intestinal microbiota in the early-life stage. The injury of the intestinal microbiota in the early-life stage may continue to the adult stage and affect growth and development and physiological metabolism.

Animals ; Animals, Newborn ; Anti-Bacterial Agents ; pharmacology ; Ceftriaxone ; pharmacology ; Female ; Gastrointestinal Microbiome ; drug effects ; Intestinal Mucosa ; drug effects ; Ki-67 Antigen ; analysis ; Mice ; Mice, Inbred BALB C ; Mucin-2 ; analysis ; Zonula Occludens-1 Protein ; analysis

Objective To investigate the effects of Ghrelin on the expression of acyl coenzyme A: cholesterol acyhransferases-1 (ACAT-1) in THP-1 derived foam cells. Methods The human monocytic leukemia cell line (THP-1) was chosen in our study. The differentiation of THP-1 cells into macrophages was induced by phorbol 12-myristate 13-acetate. Macrophages were then incubated with oxidized LDL (ox-LDL) to generate foam cells. Ghrelin and [D-Lys3]-GHRP-6, the special antagonist of growth hormone secretagogue receptor (GHS-R), were treated during foam cells formation. The ACAT-1 protein and mRNA levels were detected by Western blot and RT-PCR. The effect of variance of cholesterol content was measured by zymochemistry via-fluorospectrophotometer. Results Ghrelin reduced the content of cholesterol ester in foam cells obviously. ACAT-1 protein and mRNA levels were also decreased. The antagonist of GHS-R inhibited the effects of Ghrelin on ACAT-1 expression in dose-dependent manner. The ACAT-1 mRNA levels of the GHS-R specific antagonist groups (10~(-5), 5×10~(-5), 10~(-4) mol/L) were 1.14±0.04、1.58±0.03、 2.40±0.16, significantly higher than that of the Ghrelin group (0.89±0.05). And the protein expressions were 1.25±0.09,1.77±0.11,2.30±0.09, also higher than that of the Ghrelin group (0.86±0.08). Conclusions Ghrelin might interfere atherosclerosis by down-regnlating the expression of ACAT-1 via GHS-R pathway.