ABSTRACT
The pathogenesis and treatment strategies of acute myeloid leukemia (AML) are different for disparate gene mutations. Therefore, precise molecular testing plays a vital role in its diagnosis. However, when clinical laboratories perform molecular testing, internal quality control materials similar to clinical samples for molecular testing are lacking. At the same time, there is no related external quality assessment system to evaluate clinical laboratory test results. In order to improve the accuracy and credibility of molecular testing in clinical laboratories, we used the CRISPR/Cas9 technology to construct the DNMT3A (R882H, 2645G > A) HEK293T cell line for the quality control of AML molecular testing. We replaced the cas9 protein recognition site AGG in the ssODN with AGA to prevent the homologous recombination cell line from being cleaved by the cas9 protein again, thereby increasing the success rate of homologous recombination cell line production. It has been verified that the DNMT3A (R882H, 2645G > A) cell line can be inherited stably. The mutation frequency of the external quality assessment sample made by the DNA extraction of DNMT3A (R882H, 2645G > A) HEK293T cell line was very stable, tested by two Sanger sequencing instruments and three NGS instruments. The results above showed that the DNA extraction of DNMT3A (R882H, 2645G > A) HEK293T cell line can not only be used as internal quality control, but also be used as external quality assurance samples for monitoring different manufacturers and platforms, thereby improving the accuracy and credibility of molecular testing in clinical laboratories.
ABSTRACT
Objective:To explore evaluative value of ultrasound contrast for carotid atherosclerotic plaque angiogene-sis in patients with coronary heart disease (CHD).Methods:A total of 62 CHD patients (CHD group)and 62 healthy subjects undergoing physical examination (healthy control group)were selected.Both groups received ultra-sound contrast examination.Plaque condition was observed:its time to peak (TTP),area under the curve (AUC) and ratio of maximum peak intensity (IMAX%)were compared between two groups.Results:Compared with healthy control group,there were significant rise in detection rates of soft plaque (8.06% vs.56.45%),hard plaque (3.23% vs.25.81%)and mixed plaque (1.61% vs.17.74%),P =0.001 all;significant reduction in TTP [(22.04±3.41)s vs.(17.69±3.62)s],and significant rise in IMAX% [(50.11±4.12)% vs.(68.25±6.29)%] and AUC [(1485.62±254.13)Db/s vs.(2312.15±352.69)Db/s]in CHD group,P =0.001 all.Ultrasound con-trast indicated that carotid low echo plaque showed diffuse enhancement and carotid ulcer plaque showed linear en-hancement of ulcer boundary in CHD group,but there was no significant enhancement in carotid plaque image in healthy control group.Conclusion:Ultrasound contrast can determine the nature of atherosclerotic plaques,assess degree of angiogenesis,and provide clinical reference.
ABSTRACT
Objective:To explore evaluative value of ultrasound contrast for carotid atherosclerotic plaque angiogene-sis in patients with coronary heart disease (CHD).Methods:A total of 62 CHD patients (CHD group)and 62 healthy subjects undergoing physical examination (healthy control group)were selected.Both groups received ultra-sound contrast examination.Plaque condition was observed:its time to peak (TTP),area under the curve (AUC) and ratio of maximum peak intensity (IMAX%)were compared between two groups.Results:Compared with healthy control group,there were significant rise in detection rates of soft plaque (8.06% vs.56.45%),hard plaque (3.23% vs.25.81%)and mixed plaque (1.61% vs.17.74%),P =0.001 all;significant reduction in TTP [(22.04±3.41)s vs.(17.69±3.62)s],and significant rise in IMAX% [(50.11±4.12)% vs.(68.25±6.29)%] and AUC [(1485.62±254.13)Db/s vs.(2312.15±352.69)Db/s]in CHD group,P =0.001 all.Ultrasound con-trast indicated that carotid low echo plaque showed diffuse enhancement and carotid ulcer plaque showed linear en-hancement of ulcer boundary in CHD group,but there was no significant enhancement in carotid plaque image in healthy control group.Conclusion:Ultrasound contrast can determine the nature of atherosclerotic plaques,assess degree of angiogenesis,and provide clinical reference.