Clinical diagnostic value of procalcitonin detection in local infection and sepsis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the value of procalcitonin (PCT) detection in the diagnosis of local infection and sepsis.</p><p><b>METHODS</b>PCT, C-reactive protein (CRP), white blood cell count (WBC), neutrophil ratio (neu%) and lymphocyte ratio (lym%) were measured in patients with negative or positive blood culture test. The receiver operating characteristic (ROC) curves were constructed for PCT CRP, WBC, neu%, lym%, and the diagnostic model using SPSS software. Based on the binary logistic regression model, the predictors or probabilities were obtained and applied to establish the empirical and binormal model of the ROC curves to compare the area under the curve (AUC).</p><p><b>RESULTS</b>A highly significant difference in PCT concentrations was noted between the two groups (chi(2)=52.52, P<0.001), and the diagnostic criteria at <2 of the ROC curves resulted in the greatest Youden index with a sensitivity of 63.3% and specificity of 86.8%. The AUC of PCT, CRP, WBC, neu% and lym% were 0.700, 0.765, 0.636, 0.618 and 0.648, respectively; the combined predicted ROC AUC was 0.776. The maximum Youden index was acquired at the optimal cutoff point of 0.566 with a diagnosis sensitivity and specificity of 63.8% and 84.7%, respectively.</p><p><b>CONCLUSIONS</b>The PCT level is a valuable predictor for a rapid and reliable early diagnosis of sepsis. The diagnostic model based on the laboratory parameters, using the combined predictors of PCT, CRP and lym%, can be a useful means for predicting early-onset sepsis.</p>

Inhibition of tissue factor expression in endothelial cells by lentivirus mediated shRNA / 中南大学学报(医学版)

OBJECTIVE@#To construct the recombinant lentivirus RNAi vector, and to determine whether the lentivirus mediated short hairpin RNA (shRNA) can inhibit the tissue factor (TF) expression in endothelial cells.@*METHODS@#Two short hairpin RNAs targeting to human TF were cloned into pENTRTM/U6 plasmid to obtain an entry clone, and the positive clones were verified by sequencing. A recombination reaction was performed between the pENTR/U6 entry construction and pLenti6/BLOCKiTTM-DEST vector, and then the positive clones were confirmed by sequencing. The 293FT cell line was transfected by the above recombined plasmid and lentivirus packing materials, the culture supernatant was harvested, and the virus titer was determined. RT-PCR and ELISA were used to observe the inhibition of TF gene expression after the lentivirus transduction in human umbilical vein endothelial cells.@*RESULTS@#The shRNA sequences targeting to human TF were cloned into the vectors, and an entry clone and an expression clone were constructed successfully, which were proved by sequence determination. Viral particles were packaged in the 293FT cell line, all virus stocks were collected, and the transfection titer was 5*10(5)/transduced unit. RT-PCR and enzyme linked immunosorbent assay demonstrated that the lentivirus stocks could suppress the TF expression in endothelial cells remarkably.@*CONCLUSION@#Lentivirus RNAi vectors containing human TF gene are successfully constructed, and lentivirus mediated shRNA can inhibit the TF expression in endothelial cells, which may provide a highly effective method for the prevention and treatment of thrombo-embolic diseases.

Relationship between surface molecules and RelB gene expression in DC2.4 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between the expression of the nuclear factor-kappaB transcription factor RelB gene and the surface molecules in DC2.4 cell line.</p><p><b>METHODS</b>DC2.4 cell line was cultured in complete RPMI 1640 medium, whose morphology was observed with optical microscope and the intracellular structures with transmission electron microscope. Flow cytometry was performed to analyze the surface markers of the cells, including MHC-II, CD86 and CD40, and RelB mRNA expression was detected by RT-PCR.</p><p><b>RESULTS</b>Under optical microscope, the cells appeared irregular in shape with obvious dendritic cell processes, and electron microscopy revealed homogenous fat drops and phagocytic vesicles in the cytoplasm. Flow cytometry demonstrated low expression levels of MHC-II and CD40, but high level of CD86 molecules. Low-level expression of RelB mRNA was detected by RT-PCR, resembling its expression level in bone marrow-derived DC with immature phenotype.</p><p><b>CONCLUSION</b>DC2.4 is a mouse bone marrow dendritic cell line with strong phagocytic capacity, and the low expression of both RelB gene and surface CD40 molecules suggests an immature dendritic cell line.</p>

Relationship between polymorphisms of interleukin 10 promoter and serum levels of lipoprotein in the Chinese Han population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the relationship between polymorphisms of interleukin 10 (IL10QX) promoter and serum levels of lipoprotein in the healthy Chinese Han population.</p><p><b>METHODS</b>PCR restriction fragment length polymorphism assay was used to detect the distribution of genotypes of IL10 -592,-819,-1082 in 200 healthy Chinese Han subjects. Serum levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and very low-density lipoprotein (VLDL) in all subjects were measured to analyze the relationship with the polymorphisms of IL10 promoter.</p><p><b>RESULTS</b>Comparing with AA genotype, the group with GA genotype at IL10 promoter -1082 position had a significant elevation of serum HDL-C level [(1.514+/-0.501) mmol/L vs. (1.261+/-0.346) mmol/L, t=-2.225, P=0.028] and a lower serum TG level[(1.701+/-1.836) mmol/L vs. (0.981+/-0.314) mmol/L,Z=-2.096,P=0.036]. The TG, TC, HDL-C, LDL-C and VLDL levels did not show any statistically significant differences among different genotypes (CC, AA, CA) of the IL10 -592, as well as the genotypes (TT, TA, AA) ofIL10 -819 (P>0.05).</p><p><b>CONCLUSION</b>The results suggest that in the Chinese Han population, the polymorphism at position -1082 in the promoter region of IL10 gene may be associated with the serum HDL-C level and TG level.</p>

Heart-type fatty acid-binding protein detection for early diagnosis of acute coronary syndrome / 南方医科大学学报

<p><b>OBJECTIVE</b>To estimate the reliability of heart-type fatty acid-binding protein (H-FABP) for identifying acute coronary syndrome (ACS) in the early stage of chest pain onset.</p><p><b>METHODS</b>This investigation was conducted based on a small population consisting of 40 healthy individuals, 19 established AMI patients and 20 unstable angina pectoris (UAP) patients. Serum H-FABP concentrations were measured in these subjects by sandwich ELISA, and receiver operating characteristics (ROC) curves for H-FABP for diagnosing AMI and UAP against normal subjects were then generated respectively. The areas under curve (AUCs) were calculated, and 0.5 was defined as the critical value of AUC to evaluate the diagnostic ability.</p><p><b>RESULTS</b>The concentrations of H-FABP in healthy individuals, AMI patients and UAP patients were 1.29+/-0.64, 24.45+/-32.40 and 1.95+/-3.11 ng/ml, respectively; AUC (AMI) and AUC UAP were 0.978 (95%CI: 0.948-1.000) and 0.503 (95% CI: 0.334-0.671) respectively, and the former was significantly greater than 0.5.</p><p><b>CONCLUSIONS</b>In the early stage of chest pain onset H-FABP detection is sufficient in distinguishing AMI patients from healthy individuals, but not capable of distinguishing UAP patients from healthy individuals. H-FABP may be used as a diagnostic biochemical marker in the early stage of AMI.</p>