In this paper, a nucleic acid protein analyzer based on Lambert-Beer law and ultraviolet spectrophotometry is introduced, which is composed of ultraviolet monochromatic light generator, photoelectric signal detection module, vortex mixer, touch screen and embedded central controller. For ultra-micro measurement, a continuous-wavelength full-spectrum spectrophotometric detection circuit is designed in the hardware part. The transmitted light signal is collected by silicon photodiode, amplified and processed by subsequent circuit, and then transmitted to a single chip computer STM32F407VGT6 with CortexTM-M4 core after A/D conversion. The concentration and purity of nucleic acid protein are evaluated by assistant software detection algorithm. The instrument has the characteristics of compact size, flexible use, simple operation, high sensitivity and high detection efficiency. The experimental results show that the instrument has good sensitivity, repeatability and accuracy, and is suitable for the ultra-micro measurement of nucleic acid sample concentration, purity and protein concentration.
Algorithms ; Nucleic Acids/analysis* ; Proteins ; Software ; Spectrophotometry, Ultraviolet

The aim of this study was to build a gene chip system with surface plasmon resonance (SPR) technique, for which Gamma-peptide nucleic acid (Gamma-PNA) functioned as a probe, in order to improve sensitivity and its specificity. With the use of self-assembled monolayer (SAM) technology, surface chemistry of two-dimensional structure was used. Gamma-PNA was designed according to the bioinformatics, and was plated on the SPR chip modified by SAM. Subsequently, relevant parameters of the experiment were ensured and optimized. The results showed that the performances of Gamma-PNA probe was little affected by the ion concentration of buffer, and it had a strong light signal in a stable state. As the ion concentration was 0, there were still good hybrid reactions; pH value had less influence upon Gamma-PNA probe, and acid environment of buffer could be better. Gamma-PNA probe combined with sensor technologies achieved made the probe with dispensable labels and real-time detection. It also improved the efficiency of the hybridization and the stability, providing the foundation for clinical application.
Nucleic Acid Hybridization ; Nucleic Acid Probes ; Oligonucleotide Array Sequence Analysis ; methods ; Peptide Nucleic Acids ; genetics ; Sensitivity and Specificity ; Surface Plasmon Resonance

OBJECTIVETo develop a solid phase PCR method by covalent single point immobilization for recycle utilization of human genome.

METHODSPolymethacrylamide gel was selected as a solid PCR carrier based on DNA-hydrogel copolymer chemistry presented by Mirzabekov. (CH2)6NH2 amino-modified PCR product and randomly fractured formic acid-modified plasmid pGEM-T-HLA-G were used as templates. The specificity of the attachment chemistry was characterized by acrylamide gel electrophoresis, and the thermal stability of method was demonstrated by PCR. This method was applied for the recycle utilization of human genome. Sequencing was used to exclude the possibility of introduced mutations during modification and immobilization procedures.

RESULTSThe PCR detections of plasmid DNA and human genome DNA immobilized by polymethacrylamide gel was successful. The thermal stability of method was successfully demonstrated by performing PCR after 16 rounds of standard 36 PCR cycles. And the sequencing was found no mutation.

CONCLUSIONThe DNA immobilization method with polymethacrylamide gel as a solid phase carrier is stable and specific, which can be a possible approach for realizing recycle utilization of human genome for whole-genome sequencing and SNP detection.

Electrophoresis, Polyacrylamide Gel ; Genome, Human ; Humans ; Hydrogels ; Immobilized Nucleic Acids ; analysis ; Polymerase Chain Reaction ; methods

Non-invasive prenatal screening using cell-free fetal DNA (NIPS) has been integrated into the prenatal health care only in a short span of five years, and the guidelines provided by professional bodies have been continuously updated. The American College of Medical Genetics and Genomics has made a statement on NIPS in July, 2016, suggesting that the NIPS can replace conventional screening for Patau, Edwards and Down syndromes in a continuum of gestational age and for any maternal age, except those who are significantly obese. The scope of target diseases of NIPS are also growing. Meanwhile, pre- and post-test counseling for NIPS has put forward a greater challenge for medical professionals.
Cell-Free Nucleic Acids ; analysis ; DNA Copy Number Variations ; Female ; Humans ; Pregnancy ; Prenatal Diagnosis ; methods

PURPOSE: Raman spectroscopy is a vibrational spectroscopic technique, which is capable of providing details on the chemical composition, molecular structure and molecular interactions in cells and tissues. The primary objective of this study was to explore Raman spectroscopy for the detection of spectral changes between normal and cancer tissue in the stomach. METHODS: Tissue specimens were obtained from the resected stomach of advanced gastric cancer patients. The normal gastric and cancer tissues were harvested from the middle, lower portion of the stomach and from the tumor mass, respectively. 19 sets (antrum, body and cancer) of spectral data, with clearly defined histopathological findings, were selected in this study. FT-Raman spectroscopy (Bruker Inc., Karsruhe, Germany) was used for tissue Raman studies, with excitation at 1, 064 nm. The Raman spectra from the gastric tissue specimens were obtained with a 20 minute signal acquisition time. RESULTS: In the range 700~1, 900 cm-1, the Raman spectra of gastric antral tissue were dominated by a number of vibrational modes of biomolecules, such as proteins, lipids and nucleic acids. The Raman spectrum pattern of gastric body tissue was similar to that of the antrum, suggesting the structure and composition between the gastric antrum and body are much the same. The Raman spectra differed significantly between the normal and malignant cancer tissues, with cancers showing higher percentage signals for protein, lipid and nucleic acid compared to normal tissue (P<0.05). Difference were observed in the shapes of the Raman spectra between the normal and cancer tissues, particularly in the spectral ranges 1, 250~1, 255, 1, 330~1, 340 and 1, 440~1, 450 cm-1, which contain signals relating to protein and lipid conformations and CH2 bending mode of nucleic acids. CONCLUSION: This study demonstrates the ability of Raman spectroscopy to detect biochemical changes in malignant gastric tissue, and may become a useful adjunct to pathological diagnosis allowing guided biopsies and assessment of adequacy of resection margins.
Biopsy ; Diagnosis ; Humans ; Molecular Structure ; Nucleic Acids ; Pyloric Antrum ; Spectrum Analysis ; Spectrum Analysis, Raman* ; Stomach Neoplasms ; Stomach*

With the development of liquid biopsy technology, plasma cell-free DNA (cfDNA) becomes one of the research hotspots. Whole-genome bisulfite sequencing of plasma cell-free DNA has shown great potential medical applications such as cancer detection. However, the practical stability evaluation is still lacking. In this study, plasma cell-free DNA samples from two volunteers at different time were collected and prepared for sequencing in multiple laboratories. The library preparation strategies include pre-bisulfite, post-bisulfite and regular whole-genome sequencing. We established a set of quality control references for plasma cell-free DNA sequencing data and evaluated practical stability of blood collection, DNA extraction, and library preparation and sequencing depth. This work provided a technical practice guide for the application of plasma cfDNA methylation sequencing for clinical applications.
Cell-Free Nucleic Acids ; DNA Methylation ; High-Throughput Nucleotide Sequencing ; Humans ; Sequence Analysis, DNA ; Sulfites ; Whole Genome Sequencing

OBJECTIVETo investigate the characteristics of freshly resected laryngeal carcinoma by Fourier transform infrared spectroscopy (FTIR).

METHODSFTIR was applied to the study of the cancerous tissues and adjacent normal tissues in 32 patients.

RESULTSCompared with pathological diagnosis results, one benign specimen was classified as a malignant, the accuracy was 98.4%. Significant differences were seen in the FTIR spectra between the normal and malignant laryngeal tissues. The peak at 1085 cm(-1) shift to 1114 cm(-1) showed that the relative contents of DNA in laryngeal carcinoma cells was increased. The peak at 1397 cm(-1) was stronger than 1451 cm(-1) in normal tissues, while it was not obvious in cancer tissues. I(2926)/I(2870) in carcinoma cells was lower than that in normal tissues. The wave numbers of the bands of amide I and amide II, symmetric and asymmetric stretching bands of CH(3), stretching vibration bands of C-OH and NH band were shifted to higher number in cancer tissues.

CONCLUSIONThe study shows that the malignant and normal laryngeal tissues have different FTIR spectra, which are mainly due to changes in protein, nucleic acid and phospholipids. FTIR may become a new method for the diagnosis of laryngeal carcinoma in clinical practice.

Humans ; Laryngeal Neoplasms ; chemistry ; diagnosis ; pathology ; Larynx ; chemistry ; pathology ; Neoplasm Proteins ; analysis ; Nucleic Acids ; analysis ; Phospholipids ; analysis ; Spectroscopy, Fourier Transform Infrared ; methods

OBJECTIVETo develop a new sampling medium for detecting of bioaerosols.

METHODSThe sampling media were tested by using Escherichia coli, Staphylococcus aureus and Serratia marcescens under static and active conditions, preliminary applications were performed using AGI-10 and high volume sampler.

RESULTSThe average recovery rates were raised to 24.7%, 58.2%, 40.5%, 44.1%, 20.5%, and 15.4%, respectively in six consecutive experiments under static condition for 60 min at room temperature. Four kinds of sampling media were singled out after static experiments, which were referred to as "samplutions" PD1, PX2, TD1, and TX2, respectively. Under the active condition, the protective efficacy of PD1, PX2, TD1, and TX2 was 226% (153/47), 553% (111/17), 150% (120/48), and 268% (419/114), respectively.

CONCLUSIONThe samplutions have some effects on the subsequent nucleic acid detection, which could be avoided by employing standard nucleic acid extraction procedure. The newly developed samplution can be applied to the detection of bioaerosols.

Aerosols ; analysis ; Air Microbiology ; Air Pollutants ; analysis ; Environmental Monitoring ; methods ; Escherichia coli ; isolation & purification ; Nucleic Acids ; isolation & purification ; Sampling Studies ; Serratia marcescens ; isolation & purification ; Staphylococcus aureus ; isolation & purification

As the demand for large-scale analysis of gene expression using DNA arrays increases, the importance of the surface characterization of DNA arrays has emerged. We compared the efficiency of molecular biological applications on solid-phases with different surface polarities to identify the most optimal conditions. We employed thiol-gold reactions for DNA immobilization on solid surfaces. The surface polarity was controlled by creating a self-assembled monolayer (SAM) of mercaptohexanol or hepthanethiol, which create hydrophilic or hydrophobic surface properties, respectively. A hydrophilic environment was found to be much more favorable to solid-phase molecular biological manipulations. A SAM of mercaptoethanol had the highest affinity to DNA molecules in our experimetns and it showed greater efficiency in terms of DNA hybridization and polymerization. The optimal DNA concentration for immobilization was found to be 0.5 microM. The optimal reaction time for both thiolated DNA and matrix molecules was 10 min and for the polymerase reaction time was 150 min. Under these optimized conditions, molecular biology techniques including DNA hybridization, ligation, polymerization, PCR and multiplex PCR were shown to be feasible in solid-state conditions. We demonstrated from our present analysis the importance of surface polarity in solid-phase molecular biological applications. A hydrophilic SAM generated a far more favorable environment than hydrophobic SAM for solid-state molecular techniques. Our findings suggest that the conditions and methods identified here could be used for DNA-DNA hybridization applications such as DNA chips and for the further development of solid-phase genetic engineering applications that involve DNA-enzyme interactions.
Chimera ; DNA ; Gene Expression ; Genetic Engineering ; Immobilization ; Immobilized Nucleic Acids ; Ligation ; Mercaptoethanol ; Molecular Biology ; Multiplex Polymerase Chain Reaction ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Polymerization ; Polymers ; Reaction Time ; Surface Properties

Multiple endocrine neoplasia type 1 is an autosomal dominant disease caused by the MEN1 germline mutation. A 25-year-old male was admitted for loss of consciousness. Initial laboratory data showed hypoglycemia and hypercalcemia. The image study showed two insulinoma in the pancreas head and body. MIBI scan was positive in the left lower parathyroid gland. After diagnosis of insulinoma and hyperparathyroidism, MEN1 was suspected, but there was no family history of endocrine disease. Enucleation of the insulinoma in the pancreatic head and body was performed. After the operation, the blood sugar level was normalized and no hypoglycemic symptoms were observed. Testing of germline mutations of the MEN1 gene was performed by direct DNA sequence analysis after obtaining informed consent. In the genetic study, a frameshift mutation was found in exon 2 which deleted 16 nucleic acids (c.326_341del16) and resulted in a truncation at codon 113. This mutation was not reported previously. We found a novel and de novo mutation of the MEN1 gene. Genetic study is necessary in case of young-age, multiple endocrine tumors.
Adult ; Blood Glucose ; Codon ; Diagnosis ; Endocrine System Diseases ; Exons ; Frameshift Mutation ; Germ-Line Mutation* ; Head ; Humans ; Hypercalcemia ; Hyperparathyroidism* ; Hypoglycemia ; Informed Consent ; Insulinoma* ; Male ; Multiple Endocrine Neoplasia Type 1* ; Nucleic Acids ; Pancreas ; Parathyroid Glands ; Sequence Analysis, DNA ; Unconsciousness