ABSTRACT
Objective@#Hyperbaric oxygen treatment (HBOT) has demonstrated efficacy in improving hearing levels of patients with idiopathic sudden sensorineural hearing loss (ISSHL); however, the underlying mechanisms are not well understood. HBOT alleviates the inflammatory response, which is mediated by Toll-like receptor (TLR) 4 and nuclear factor (NF)-κB. In this study we investigated whether HBOT attenuates inflammation in ISHHL patients alteration of TLR4 and NF-κB expression.@*Methods@#ISHHL patients ( = 120) and healthy control subjects ( = 20) were enrolled in this study. Patients were randomly divided into medicine group treated with medicine only ( = 60) and HBO group receiving both HBOT and medicine ( = 60). Audiometric testing was performed pre- and post-treatment. TLR4, NF-кB, and TNF-α expression in peripheral blood of ISSHL patients and healthy control subjects was assessed by ELISA before and after treatment.@*Results@#TLR4, NF-κB, and TNF-α levels were upregulated in ISSHL patients relative to healthy control subjects; the levels were decreased following treatment and were lower in the HBO group than that in the medicine group post-treatment ( < 0.05 and < 0.01).@*Conclusion@#HBOT alleviates hearing loss in ISSHL patients by suppressing the inflammatory response induced by TLR4 and NF-κB signaling.
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , China , Hearing Loss, Sensorineural , Therapeutics , Hearing Loss, Sudden , Therapeutics , Hyperbaric Oxygenation , Inflammation , Genetics , Therapeutics , NF-kappa B p50 Subunit , Genetics , Metabolism , Toll-Like Receptor 4 , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To establish a comprehensive and simple assay using denaturing high performance liquid chromatography (DHPLC) for the diagnosis of most common mutations and deletions of α-thalassemia gene in Southeast Asians and Southern Chinese.</p><p><b>METHODS</b>This assay has included a duplex polymerase chain reaction (PCR) followed by DHPLC analysis. An improved PCR was also performed followed by DHPLC analysis. With this assay, a blinded study of 160 samples was screened for three common mutations and three common deletions.</p><p><b>RESULTS</b>The duplex PCR-DHPLC combined with the improved PCR-DHPLC analysis has detected all mutations and the wild-type allele. The results were consistent with those by the original methods.</p><p><b>CONCLUSION</b>This molecular assay may be used for the diagnosis of α-thalassemia patients from this geographical region. The method is accurate, rapid, semi-automatic and cost-effective, which makes it suitable for large-scale screening.</p>
Subject(s)
Humans , Chromatography, High Pressure Liquid , Methods , DNA Mutational Analysis , Methods , Gene Order , Genotype , alpha-Globins , Genetics , alpha-Thalassemia , Diagnosis , GeneticsABSTRACT
To construct a safer and more efficient gene engineering Lactococcus Lactis for expressing phenylalaine ammonia lyase (PAL) which will be benefit for PKU therapy, pal cDNA of Parsly and synthesized sequence based on Lactococcus Lactis bias codons were recombined into two Lactococcus Lactis NICE systems. The activities of the expressed PAL were detected, and the effect of Lactococcus Lactis bias codons on the expression of exterior protein was analyzed. The results showed that the expression level of PAL was increased by using Lactococcus Lactis bias codons in both Lactococcus Lactis NICE systems. Through which several safer andmore efficient strains of the gene engineering Lactococcus Lactis were obtained.
Subject(s)
Cloning, Molecular , Codon , Genetics , Genetic Vectors , Genetics , Lactococcus lactis , Genetics , Metabolism , Phenylalanine Ammonia-Lyase , Genetics , Recombinant Proteins , Metabolism , Transformation, BacterialABSTRACT
To construct a new high effective genetic engineering strain which can express active PAL enzyme in Lactococcus lactis (L.L), and acquire better effect on curing hyperphenylalaninemia rats, Firstly translational fusion vector and transcriptional fusion vector were constructed in E. coli MC1061, and then PAL cDNA was transformed into L.L. Two kinds of high effect strain were compared with their enzyme activity and animal experiment was carried out. The results showed: (1) Two kinds of engineering L.L. were obtained and translational fusion strain has higher level enzyme activity. (2) The amount of transcinnamic aicd reach peak when induced for 6 hours. (3) The blood phe level of the treated rats was significantly reduced compared with non-treated rats when receiving fresh p(NZ8048-PAL)1/NZ9000. The engineering L.L(translational fusion strain) can significantly reduce the blood phe level of the hyperphenylalaninemia rats, which has more superiority than pMG36e-PAL/L. L.