The basic principle of high resolution melting ( HRM ) is that DNA -saturated fluorescent dye is added into the PCR reaction system;the PCR amplicon is denatured by heating in a certain temperature range;specialized instrumentsdetect degeneration double-stranded DNA fluorescence signal and draw the melting curve and then wild-type and heterozygous mutants are differentiatedaccording to different melting curve shape .HRM technology is widely used in gene mutation scanning because of its advantages of simplicity, accuracy, fastness, low cost, short cycle time and high throughput .In addition, compared with other mutation screening methods , HRM analysis is a real closed-tube method , which indicates that high-resolution melting analysis is directly performed after PCR reaction and HRM technology is more convenient . In conclusion , HRM technology is a new technology for mutation screening and genotyping .

With the development of the concept of precision medicine, under the background of the new coronavirus pneumonia epidemic, the clinical diagnosis and treatment of infectious diseases has received more and more attention. The experimental diagnosis technology with molecular biology as the core is used as important means for the clinical laboratory diagnosis of infectious diseases. This lcind of technology is paid special attention. In recent years, advances in nanomaterials, applied chemistry, photophysics, and biosensing technologies have also ushered in revolutionary and creative developments in molecular diagnostic technology. This article reviews the application and development of the latest molecular diagnostic technologies, such as next-generation quantitative PCR technology and gene sequencing technology, isothermal amplification technology, biochip and biosensor technology in the clinical diagnosis of infectious diseases.

With the advancement of medical science, point-of-care testing (POCT), which is simple to operate, quick to respond, and does not rely on equipment and professional and technical personnel, is expected to realize continuous monitoring, diagnosis, management and screening of patients. It is an important development direction of the in vitro diagnostic industry. Combining the principles of new technology, POCT is inevitably developing towards the transformation from qualitative to precise quantitative under the premise of improving sensitivity and specificity. During the development of the POCT platform, the simple, fast, low-cost loop-mediated isothermal amplification (LAMP) technology with high-efficiency amplification characteristics plays an increasingly important role. This article summarizes the mechanism of the LAMP technology, the research progress, and the clinical application of the POCT platform based on the LAMP. The current deficiencies and future development directions of the POCT platform based on the LAMP are also discussed.

Cultivating comprehensive personnel with competent professional ethics, technical skills and scientific research capabilities is the goal and task of today's laboratory medical education. How to make full use of diversified teaching materials, tools and methods to improve teaching quality is worthy of exploration and thinking in laboratory medical teaching units. In this paper, using mind map as a tool, taking the COVID-19 epidemic as an example, we intend to design a set of multidisciplinary and all-round integrated curriculum of medical laboratory education, and discuss the application status and prospect of integrated curriculum and mind map in medical laboratory education. Through doing this, we aim to optimize the training goals of laboratory medicine education, innovate the teaching methods and boost the training efficiency of laboratory medicine education to keep up with the times.

OBJECTIVE: To assess the influence of FLT3 expression on the prognosis of patients with acute myeloid leukemia (AML) by cell experiment and clinical data analysis. METHODS: Models for FLT3 over-expression and interference-expression in AML cells were constructed. The level of BAK gene expression and its protein product was determined, along with the proliferation and apoptosis of leukemia cells. FLT3 gene expression and FLT3-ITD variant were determined among patients with newly diagnosed AML. RESULTS: Compared with the interference-expression group, the level of BAK gene expression and its protein in FLT3 over-expression AML cells was significantly lower (P < 0.001), which also showed significantly faster proliferation (P < 0.001) and lower rate of apoptosis (P < 0.001). The expression level of FLT3 gene among patients with newly diagnosed AML was also significantly higher compared with the healthy controls (P < 0.001). The FLT3 gene expression of FLT3-ITD positive AML patients was higher than that of FLT3-WT patients (P = 0.002). Survival analysis showed that AML patients with high FLT3 expression in the medium-risk group had a lower complete remission rate and overall survival rate compared with those with a low FLT3 expression (P < 0.001). CONCLUSION Over-expression of FLT3 may influence the course of AML by promoting the proliferation of leukemia cells and inhibiting their apoptosis, which in turn may affect the prognosis of patients and serve as a negative prognostic factor for AML.
Humans ; Apoptosis/genetics* ; Data Analysis ; Leukemia, Myeloid, Acute/genetics* ; Gene Expression ; fms-Like Tyrosine Kinase 3/genetics*

Objective To observe the changes of liver and kidney function in hepatitis B virus(HBV)carriers after renal transplantation. Methods A total of 116 patients with HBV infection undergoing renal transplantation and 348 counterparts without HBV infection were recruited in this clinical trial. The liver function parameters including alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and renal function parameter including serum creatinine(Scr)level were measured before and at 1, 3, 6, 12, 18, 24, 36 and 60 months after renal transplantation. Preoperative and postoperative changes of liver and kidney function were statistical y compared between the hepatitis B surface antigen(HBsAg)(+)and HBsAg(–)groups. According to the results of preoperative HBV serology, preoperative quantitative detection of HBV DNA and preoperative liver function test, 116 HBsAg(+)patients undergoing renal transplantation were divided into(HBsAg, HBeAg and anti-HBc all positive)and(HBsAg, anti-HBe and anti-HBc all positive)groups, HBV DNA(+)and HBV DNA(–)groups, and normal and abnormal liver function groups.Preoperativeandpostoperativechangesofliverandkidneyfunctionwerestatisticalycomparedbetweendifferentsubgroups. Results(1)Preoperative ALT and AST levels in HBsAg(+)patients were significantly higher compared with those in their HBsAg(–)counterparts. In 36 months after renal transplantation, liver function parameters significantly differed between two groups(al P<0.05), whereas no statistical significance was noted at postoperative 60 months(al P>0.05). Before and in 60 months after renal transplantation, no statistical significance was observed in the Scr levels between the HBsAg(+)and HBsAg(–)groups(all P>0.05).(2)Before and in 60 months after renal transplantation, no statistical significance was observed in the liver and kidney function parameters between the(HBsAg, HBeAg and anti-HBc all positive)and(HBsAg, anti-HBe and anti-HBc all positive)groups, and HBV DNA(+)and HBV DNA(–)groups(all P>0.05).(3)The ALT levels before and at 1, 3, 6 and 12 months after renal transplantation significantly differed between the normal and abnormal liver function groups(al P<0.05), whereas no statistical significance was observed at other time points(all P>0.05). The AST levels before and at 1 month after renal transplantation significantly differed between two groups(both P<0.05), whereas did not significantly differ at alternative postoperative time points(all P>0.05). No statisticalsignificancewasobservedinthekidneyfunctionparametersbeforeandat60monthsfolowingrenaltransplantation between two groups(al P>0.05). Conclusions HBV infection cannot exert significant effect upon kidney function within 5 years after renal transplantation, whereas it can affect short-term postoperative liver function.

The detection technology based on micro-nanofluidics has been developed fast with wide applications in biochemical analysis and clinical diagnostics because of its small sample volume requirement, rapid detection and portability. Recently, nanofluidics emerges for DNA sequencing and biomarker detection. With nanopore technology,whole genome sequence in 24 hours with a cost of lower than $1000 could be realized. However, improvement in the detection accuracy and repeatability are still desired for clinic diagnostics. Nanopore technology could be powerful tools for clinical diagnostics, providing new opportunities for laboratory medicine.

During the outbreak of coronavirus disease-19 (COVID-19), the clinical laboratories of hospitals designated for the disease treatment is undertaking a lot of clinical testing work of infectious specimens. How to manage the biosafety risk is a major problem that the clinical laboratory and the nosocomial infection control department are facing. This article introduces the hierarchical prevention and control biosafety measures in the clinical laboratory from the perspective of the laboratory, with a view to provide reasonable and feasible methods for the clinical laboratories of hospitals at various levels during the outbreak.

Adult ; China ; DNA Repeat Expansion ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; RNA, Long Noncoding ; genetics ; Spinocerebellar Degenerations ; genetics ; physiopathology

Tongue squamous cell carcinoma is highly malignant and has a poor prognosis. In this study, we aimed to combine whole-genome sequencing, whole-genome methylation, and whole-transcriptome analyses to understand the molecular mechanisms of tongue squamous cell carcinoma better. Oral tongue squamous cell carcinoma and adjacent normal tissues from five patients with tongue squamous cell carcinoma were included as five paired samples. After multi-omics sequencing, differentially methylated intervals, methylated loop sites, methylated promoters, and transcripts were screened for variation in all paired samples. Correlations were analyzed to determine biological processes in tongue squamous cell carcinoma. We found five mutated methylation promoters that were significantly associated with mRNA and lncRNA expression levels. Functional annotation of these transcripts revealed their involvement in triggering the mitogen-activated protein kinase cascade, which is associated with cancer progression and the development of drug resistance during treatment. The prognostic signature models constructed based on WDR81 and HNRNPH1 and combined clinical phenotype-gene prognostic signature models showed high predictive efficacy and can be applied to predict patient prognostic risk in clinical settings. We identified biological processes in tongue squamous cell carcinoma that are initiated by mutations in the methylation promoter and are associated with the expression levels of specific mRNAs and lncRNAs. Collectively, changes in transcript levels affect the prognosis of tongue squamous cell carcinoma patients.
Humans ; Biomarkers, Tumor ; Nerve Tissue Proteins ; Prognosis ; Squamous Cell Carcinoma of Head and Neck/pathology* ; Tongue Neoplasms/pathology*