Clinical application of real-time fluorescence quantitative PCR for the detection of fecal tumor M2-pyruvate kinase in colorectal cancer patients / 国际检验医学杂志

Objective To investigate the application valve of real-time fluorescence quantitative polymerase chain reaction(RT-PCR) for the detection of tumor M2-pyruvate kinase(tM2-PK) DNA in patients with colorectal cancer(CRC).Methods Fragment of tM2-PK DNA(162 bp) was amplified and inserted into PGM-T vector to construct recombinant plasmid,which was used to develop RT-PCR method.Sensitivity,specificity and repeatability of RT-PCR for the detection of tM2-PK were analyzed.From Jan.2014 to Jun.2016,200 CRC patients and 100 healthy subjects were enrolled and detected for fecal and serum tM2-PK DNA by using RT-PCR,and the detected results were compared with those detected by using enzyme linked immunosorbent assay(ELISA).Results Recombinant plasmid was successfully constructed,which was certified by sequencing.The sensitivity of RT-PCR for the detection of tM2-PK DNA was 10 copy/mL,with high specificity and 0.3%-2.9% of coefficient of variation.In patients,the positive rate of fecal tM2-PK DNA,detected by RT-PCR,was 92.50%,and that of ELISA to detect tM2-PK was 80.00%.Fecal and serum levels of tM2-PK were correlated with the pathologic stages of tumour.Conclusion Self-established RT-PCR could be specificity and sensitivity for the detection of fecal tM2-PK,which could be used for the early diagnosis of CRC.

Application of quantum dots immune fluorescent labelling in colorectal cancer tissues / 国际检验医学杂志

Objective To investigate different antigens detected by a novel labelled reagent‐quantum dots(QDs) in the colorectal cancer tissues microarray(TMA) .Methods Depend on QDs streptavidin conjugate(QDs‐SA) combined specially with biotinylation IgG ,immune of luorescent histochemistry was utilized to examine expression of K‐ras ,matrix‐remodeling associated 5(MXRA5) proteins in the colorectal cancer TMA ,where the protein accurate location was observed .Results K‐ras ,matrix‐remodeling associ‐ated 5(MXRA5) proteins were high expressed in colorectal cancer tissue and located accurately in the cell membrane and nucleus of colorectal cancer cells ,respectively .Conclusion QDs exhibit excellent photostability ,broad emission spectrum and long fluorescence lifetime .Modified with streptavidin could accurately detect different protein locations in the colorectal cancer TMA .This is a novel approach for studying targeted imaging of colorectal cancer in vivo and vitro clinical diagnosis .

Effect of genetic polymorphism on disposition of calcineurin inhibitors in solid organ transplantation / 中南大学学报(医学版)

Calcineurin inhibitors, tacrolimus and cyclosporine, characterized by a narrow therapeutic index and a high variability in pharmacokinetic behaviors, are 2 basic immunosuppressive drugs widely used in solid organ transplantation. Tailoring of immunosuppressive drug therapy to the specific requirements of individual patients to optimize the efficacy and minimize the toxicity remains one of the biggest challenges for doctors in solid organ transplantation. Pharmacogenetic and pharmacogenomic researches, studying the effect of genetic polymorphism encoding drug metabolizing enzymes, drug transporters and pharmacological target molecules on drug disposition and action, hold promise to produce useful clinical tools for individualizing immunosuppressive therapy.

Effects of CYP3A5 * 3 polymorphisms on tacrolimus pharmacokinetic in renal transplantation recipients by PyrosequencingTM / 中华器官移植杂志

Objective To study the influence of CYP3A5 genetic polymorphism on tacrolimus metabolism in renal transplantation recipients and evaluate the methods of detecting CYP3A5* 3 polymorphism. Methods Ninety-seven renal recipients receiving the triple immunosuppressive (tacrolimus + mycophenolate mofetil + prednison) were genotyped for CYP3A5* 3 polymorphisms by the Pyrosequencing TM assays. Tacrolimus trough concentration of the patients was measured by enzyme multiplied immunoassay technique, and concentration/adjusted dose ratio (C/D) was investigated at the 7th day, 14th day, 1st month, 3rd month and 6th month after renal transplantation. Results The Pyrosequencing TM assays allowed for quick and accurate detection of CYP3A5* 3 genotypes. There were CYP3A5* 1/* 1 genotype in 17 cases (17. 5%), * 1/* 3 in 35cases (36. 1 %) and *3/* 3 in 45 cases (46.4 %) identified in 97 renal recipients. The C/D of CYP3A5 * 1/* 1 and * 1/* 3 patients was significantly lower than that of * 3/* 3 patients (P<0. 01)after renal transplantation. Conclusion The CYP3A5* 3 polymorphisms are significantly correlated with tacrolimus pharmacokinetic in renal transplant recipients. Detecting the CYP3AS* 3 genotype of the recipient before the transplantation by the Pyrosequencing TM assays can be used to help determine the optimal initial dosage after transplantation, resulting in individualized drug therapy.