RESUMO
BACKGROUND: The Rh blood group antigens are encoded by the RHD and RHCE genes, which possess a remarkable degree of polymorphism owing to their high homologous structures. These variants of the RH genes can lead to absence or weak expression of antigens. METHODS: Analysis of RHCE genotyping by Polymerase Chain Reaction (PCR-SSP) method specific to detect c.48G, c.48C, 109 bp insertion of IVS2, c.201A and c.307C and RhCE phenotyping, were conducted in 316 Chinese patients in previous study. One patient with discrepancy typing result was collected for further RhCE serologic typing using microcolumn gel method and tube method in saline using monoclonal antibodies. PacBio sequencing was performed for RHCE, RHD and RHAG complete sequence analysis. 3D molecular models of the protein with the wild-type and mutant residue were generated using the DynaMut web server. The effect of the mutation on the protein function was predicted by PolyPhen-2 software. RESULTS: One male patient of Chinese Han was detected with RHCE*C allele showed by PCR-SSP method but ccEE phenotype. Further PacBio sequencing identified one normal RHCE*cE allele and one RHCE*Ce allele carried a novel c.829G > A (p.Gly277Arg) variant, which the encoded amino acid located in the ninth transmembrane segment of RhCE protein. Crystallisation analysis of 3D molecular models revealed that the substitution at Arg277 leads to the formation of additional hydrogen bonds, including weak hydrogen bonds between multiple atoms. It also results in hydrophobic ion interactions between Arg277 and Ala244. This mutation is predicted to have a damaging effect on protein function. CONCLUSION: One novel RHCE*Ce allele with c.829G > A (p.Gly277Arg) variant was identified to resulting in the absence or weak expression of C and e antigens.
RESUMO
【Objective】 To explore the prognosis of critically ill patients with coagulation dysfunction using thrombelastogram(TEG) and coagulation four items combined with APACHEⅡ score. 【Methods】 From March 2017 to March 2020, 287 critically ill patients with coagulation dysfunction in our hospital were selected as the study group, and 303 patients with normal coagulation function during the same period were set as the control. The study group was divided into low-risk group(group A), intermediate-risk group(group B) and high-risk group (group C) based on the APACHEⅡ score, and into survival group and death group according to the prognosis. The difference of TEG, coagulation four items, and APACHEⅡ scores between the two groups were analyzed. The correlation and difference between TEG, coagulation four items and APACHE II score in the study group were analyzed. The ROC curve was drawn to analyze the prognostic predictive value of research indicators. 【Results】 Blood coagulation function related indicators in the study group fluctuated significantly: in comparison to the control, the CI value, MA value, and α angle were smaller, while the K time and R time were longer; among the coagulation four items, PT, APTT and TT were higher; Fg level was lower, and the APACHE Ⅱ score was higher(P0.05). There were significant differences between the TEG and coagulation function related index levels in patients with different prognosis. Compared with the survivals, the CI value, MA value and α angle of the dead group were smaller, while the K time and R time were longer; and among the coagulation four items, PT, APTT, and TT were higher, the Fg level was lower, and the APACHEⅡ score was higher (PP4\\P5>APACHE Ⅱ score>P1>P2. 【Conclusion】 TEG, coagulation four items, and APACHE Ⅱ score can be used to assess the severity of patients with severe coagulation dysfunction. and the combined application of the 3 indicators are of high value in predicting the prognosis of such patients, and can provide reference for clinical formulation or adjustment of intervention programs to correct coagulation dysfunction.