ABSTRACT
Objective To evaluate the value of procalcitonin(PCT) and endotoxin(ET) in early diagnosis and prognosis in critically illness patients with sepsis.Methods A total of 104 cases with suspected sepsis were prospectively collected in this study from February 2015 to October 2016 in ICU.There were 31 cases of systemic inflammatory response syndrome (SIRS group),30 cases of sepsis 24 cases of severe sepsis and 19 cases of septic shock.Prognosis was observed in patients with sepsis discharged,57 cases were survival (survival group) while 16 cases were dead (death group).The level of PCT and ET and the clinical characteristics were recorded,The the ROC curve was applied to evaluate the discriminative power of variables.Results The levels of PCT,APACHE-Ⅱ score,ET of cases with sepsis,severe sepsis and septic shock were significantly higher than those in SIRS group(P0.05).ROC curve showed that AUC of PCT was 0.867,which was significantly higher than that for APACHE-Ⅱ score (AUC=0.762) (P>0.05).Conclusion Compared with APACHE-Ⅱ score and ET,PCT can help clinicians to early diagnose and treat cases with sepsis.
ABSTRACT
Objective To establish a double TaqMan real-time fluorescence nested PCR method for rapid detection of α-thalassemia SEA deletion.Methods One hundred blood samples for thalassemia screening were collected from May to July of 2010 in the Tianhe Maternal and Child Health Hospital of Guangzhou.Seven fetal specimens for prenatal diagnosis were collected from December 2010 to February 2011 in Dongguan TungWah Hospital(2 villi and 5 amniotic fluid specimens).Fifty samples of α-thalassemia SEA deletion with genotyping results were selected from the sample bank of our laboratory.The double TaqMan real-time fluorescence nested PCR was applied to detect the truncated sequence of SEA deletion and the normal sequence within deletion range simultaneously for all these samples with the same detecting system.The genotype of α-thalassemia SEA deletion was accurately acquired according to the positive result of fluorescent PCR combined with the Ct value difference.Meanwhile,the accuracy and feasibility of this method were verified and analyzed by parallely detecting these samples with routine gapPCR for α-thalassemia SEA deletion.The genotype could be obtained according to PCR amplification and agarose gel electrophoresis.Results Two amplification efficiencies of the optimized dual TaqMan real-time fluorescence nested PCR system established in this study were both close to 100% with the slops of -3.153 and -3.182,respectively.The results of 50 samples of α-thalassemia SEA deletion with genotyping results showed that this method could not only realize rapid diagnosis,but also effectively avoid false negative or false-positive misdiagnosis by accurately determine the external contamination in the sample.Among 100 blood samples,eleven samples with SEA deletion were detected respectively and the diagnosis coincidence rate of these two methods was 100%,3 samples with SEA deletion were detected by gap-PCR,but 2 samples with SEA deletion and 1 villi sample with normal genotype but contaminated by SEA were detected by this method among 7 fetal samples.Conclusions A double TaqMan real-time fluorescence nested PCR method for α-thalassemia SEA deletion was developed.The method is a rapid and reliable test with simple operation,and is suitable for large-scale population screening and routine molecular diagnosis.
ABSTRACT
Objective To investigate whether conventional protein kinase C (cPKC ) βⅡ-interacting collapsin response mediator protein-2 (CRMP-2) provides neuroprotection against cerebral ischemic (I) injuries. Methods Male BALB/c mice were randomly divided into normoxic control (Nor) , HPC, Nor + Sham, HPC + Sham, Nor + I and HPC + I groups (n = 6 per group). Using our HPC and MCAO mouse models, we applied immunoprecipita-tion, two-dimensional electrophoresis and mass spectrometry to characterize cPKCβⅡ-interacting proteins and combined with SDS-PAGE and Western blot to quantitatively analyze CRMP-2 phosphorylation and degradation levels in the brain of mice after HPC and MCAO. Results The expression level of 10 cPKCβⅡ-interacting proteins changed obviously in cerebral cortex of HPC mice when compared with Nor group. One of these proteins, CRMP-2 protein level increased in particulate fraction and decreased in cytosolic fraction of cerebral cortex of HPC mice. CRMP-2 phosphorylation level in ischemic core (Ic) of cerebral cortex decreased significantly ( P < 0. 05 , n = 6) as compared with that of Nor + sham group, but CRMP-2 phosphorylation level in HPC +I group increased significantly as compared with that of Nor +I group ( P < 0. 05, n = 6). In ischemic cortex, CRMP-2 degradation (proteolysis) was observed as the appearance of 55 ku breakdown products (BDP). However, the CRMP-2 degradation level, BDPs products decreased significantly in penumbra ( P) of ischemic cortex from HPC +I group when we compared with that of Nor +I group (P < 0. 05, n = 6 ). Conclusion CRMP-2 is involved in attenuating the decrease of CRMP-2 phosphorylation in ischemic core and in inhibiting its degradation in penumbra of cerebral cortex of mice thereby to lessen the ischemic injuries.