Ear Diseases ; congenital ; diagnosis ; surgery ; Ear, External ; abnormalities ; Ear, Middle ; abnormalities ; Humans

Biomedical Research ; Humans ; Multicenter Studies as Topic ; Vestibular Diseases ; diagnosis ; therapy

Evidence-Based Medicine ; Humans ; Tinnitus ; diagnosis ; Vertigo ; diagnosis

Humans ; Vertigo

Audiology ; Deafness ; genetics ; Humans ; Otolaryngology ; trends

Anthropometry ; Female ; Humans ; Male ; Postural Balance ; Vision, Ocular ; Young Adult

Objective To discuss the probability of extracting the effective part of Yinhuang prescription with membrane separation method. Methods To extract and concentrate Yinhuang decoction with amb-class UF and first class NF, baicalin and chlorogenic acid were detected with HPLC, and polysaccharide with anthrone agent colorimetry. Results The content of polysaccharide in final membrane fraction was 3.41 mg/mL. The diversion ratio of baicalin and chlorogenic acid from primal decoction was 77.24%, 79.02%. Conclusion The effective part of Yinhuang decoction can be extracted with membrane separation method. Multiple active components including polysaccharide were retained and the method is simple.

Objective To optimize the water extractive process of Guben Zhike Electuary. Methods To arrange experiments with uniform design for optimizing suitable extractive proces, with the content of polysaccharide and astragaloside Ⅳ as the detecting indexes, inspecting the extacting times, lasting time and additional water which would affect the extraction rate of the two effective components. Result The best process was that water as extraction solvent, extracted for three times with two hours each time, and the amount of water added up to nine times of herbs. Conclusion The best process could extract most of the effective components out of the herbs, which is scientific.

To understand the structure of the soluble region of Enterovirus 68 3A protein, we construct a prokaryotic expression vector expressing the soluble region of EV-D68 3A protein, and identify the forms of expression product after purification. The EV-D68 3A(1-61) gene was amplified by PCR and then cloned into the expression vector pET-28a-His-SUMO. The recombinant plasmid was transformed into Escherichia coli BL21 induced by IPTG to express the fusion protein His-SUMO-3A(1-61). The recombinant protein was purified by Ni-NTA Agarose and cleaved by ULP Protease to remove His-SUMO tag. After that, the target protein 3A(1-61) was purified by a series of purification methods such as Ni-NTA, anion exchange chromatography and gel filtration chromato- graphy. Chemical cross-linking reaction assay was taken to determine the multiple polymerization state of the 3A soluble region. A prokaryotic expression vector pET28a-His-SUMO-3A(1-61) expressing the solution region of EV-D68 3A was successfully constructed and plenty of highly pure target proteins were obtained by multiple purification steps . The total protein amount was about 5 mg obtained from 1L Escherichia coli BL21 with purity > 95%. At the same time, those results determined the homomultimer form of soluble 3A construct. These data demonstrated that the expression and purification system of the soluble region of 3A were successfully set up and provide some basic konwledge for the research about 3A crystal structure and the development of antiviral drugs targeted at 3A to block viral replication.
Amino Acid Sequence ; Enterovirus D, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Models, Molecular ; Molecular Sequence Data ; Protein Structure, Tertiary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics ; metabolism