Patients with advanced hepatocellular carcinoma (HCC) often have poor prognosis, and early diagnosis and effective treatment measures can significantly improve the survival rate of patients. At present, alpha-fetoprotein (AFP) is the most widely used serum marker for the diagnosis of HCC in the world; however, no increase in AFP was found in some HCC patients. This article analyzes the application potential of AFP, hepatitis B core-related antigen, liquid biopsy technology, microRNA, long non-coding RNA, and exosomes in the early diagnosis of HCC and reviews related research advances, so as to provide a basis for exploring new methods for the early diagnosis of HCC.

The incurable chronic infection caused by hepatitis B virus (HBV) is the major health burden worldwide. Covalently closed circular DNA (cccDNA) exists in the nucleus of infected cells as a stable minichromosome, and when a new therapy realizes inactivation or eliminates persistent cccDNA in infected hepatocytes, the natural process of chronic infection and long-term antiviral therapy will no longer exist. This article introduces the methods targeting cccDNA, such as gene editing and epigenetic modification, so as to achieve the complete cure of HBV infection.

UNLABELLEDWe designed a pair of specific primers and a TaqMan fluorescent probe targeting the toxR gene of Vibrio parahaemolyticus (VP). After optimizing the conditions, the specialty, sensitivity and reproducibility of the detection method were evaluated.

RESULTS(1) the developed real-time PCR assay protocol detected only VP and was not affected by other normal food pathogens such as Staphylococcus aureus, Salmonela, Listeria monocytogenes. (2) the limit of detection was 25 copies of toxR gene in the detected samples, and the sensitivity of pure cultures and simulated food samples was 21 cfu/mL and 210 cfu/g. (3) the developed protocol of real-time PCR assay had a high reproducibility, and the sample's variation was 0.9% and 1.3% within the same sample and between tests. (4) the standard curve had a good linearity when the gene quantity was between 2.5x10(1) and 2.5x10(6) copies. The developed detection assay targeting the toxR gene can quantitatively detect VP in only 3 hours, and thus is an efficacious method for the detection of Vibrio parahaemolyticus.

Bacterial Proteins ; genetics ; DNA-Binding Proteins ; genetics ; Fluorescent Dyes ; metabolism ; Gene Targeting ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Taq Polymerase ; metabolism ; Transcription Factors ; genetics ; Vibrio parahaemolyticus ; genetics ; isolation & purification

We formulated a selective enrichment broth (SVV) for simultaneous growth of Salmonella, Vibrio parahaemolyticus, and Vibrio cholerae by single factor experiment and response surface method. We evaluated the enrichment effect of SVV by conventional culture method and real-time PCR assay. We obtained the SVV broth by supplementting the Buffered Peptone Water (BPW) with bile salt no. 3, potassium tellurite, and sodium citrate as inhibitors, and glucose, mannitol, snhydrous sodium sulfite and sodium pyruvate as accelerants. We also modified the concentration of sodium chloride in BPW. When mixed at equal or varied proportions, the target pathogens had a great accumulation (10(5)-10(8) CFU/mL) after incubated in SVV for 18 h at 37 degrees C with shaking. It can also effectively inhibit the competitive microflora. We detected 10 artificial simulated samples and 608 real samples using SVV with real-time PCR. After enriched in SVV for 18 h, the quantity of the bacteria in samples were above the detection limit. The SVV with PCR assay showed higher tested positive (4.06%) compared to that of the conventional detection method (3.78%) and there was no false report. In summary, SVV is a promising new multiplex selective enrichment broth that can be used in detection of seafood.
Culture Media ; Food Microbiology ; Salmonella ; growth & development ; Vibrio cholerae ; growth & development ; Vibrio parahaemolyticus ; growth & development

OBJECTIVETo detect potential mutation of iduronate-2-sulfatase (IDS) gene in a family affected with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).

METHODSFor the proband and his unaffected mother, the whole coding sequence of the IDS gene was analyzed with PCR and bidirectional Sanger sequencing.

RESULTSA novel splicing mutation, c.709-1G>A, was detected in the proband, for which his mother was heterozygous.

CONCLUSIONThe c.709-1G>A splicing mutation of the IDS gene is probably causative for the MSP Ⅱ in the proband. Prenatal diagnosis for the mutation may avoid birth of further child affected with this disease.

Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Glycoproteins ; genetics ; metabolism ; Heterozygote ; Humans ; Iduronate Sulfatase ; genetics ; metabolism ; Male ; Mothers ; Mucopolysaccharidosis II ; diagnosis ; enzymology ; genetics ; Mutation

OBJECTIVETo analyze mutations of IDUA gene in two pedigrees affected with mucopolysaccharidosis type I and provide prenatal diagnosis for them.

METHODSThe 14 exons of the IDUA gene were subjected to PCR amplification and Sanger sequencing.

RESULTSFor pedigree 1, the proband was found to harbor compound heterozygous mutations c.46-57delTCGCTCCTGGCC (p.Ser16_Ala19del) of exon 1 and c.1147delC (p.Arg383Alafs*57) of exon 8 of the IDUA gene, which were inherited from his father and mother, respectively. The latter was unreported previously. Prenatal diagnosis suggested that the fetus has carried a heterozygous c.46-57delTCGCTCCTGGCC mutation. For family 2, the proband was also found to carry compound mutations of the IDUA gene, namely c.721T to C (p.Cys241Arg) of exon 6 and c.1491delG (p.Thr497fs27) of exon 8, which were inherited from her mother and father, respectively. Neither mutation was reported previously. Prenatal diagnosis suggested that the fetus has carried a heterozygous c.721T to C mutation.

CONCLUSIONMutations of the IDUA gene probably underlie the MPS-I in both pedigrees. Above results have enriched the spectrum of IDUA gene mutations and facilitated prenatal diagnosis for both families.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; China ; DNA Mutational Analysis ; Female ; Fetal Diseases ; diagnosis ; genetics ; Heterozygote ; Humans ; Iduronidase ; genetics ; Male ; Molecular Sequence Data ; Mucopolysaccharidosis I ; diagnosis ; embryology ; genetics ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Sequence Deletion