Agricultural Workers' Diseases ; Humans ; Musculoskeletal Diseases

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

Xanthan gum is a microbial, natural high molecular weight polysaccharide produced by a the bacterium Xanthomonas campestris. Due to its exceptional rheological properties, its numerous areas of application cover a broad range. This review focuses on various aspects of xanthan production, properties, degradation, and application.

Objective To study and assess clinical application of two Nickel-Titanium(NiTi)rotary instru- ments,namely ProTaper and Hero 642,in preparation of root canals.Methods 125 teeth were divided into three groups and respectively instrumented by stainless K-files,ProTaper or Hero 642 rotary instruments.All teeth were obturated with lateral condensation method.The efficiency of preparation and obturation was analyzed with radio- graphs.Results NiTi rotary instruments were better in keeping the curvature and flow of curved canals than stain- less K files.There was no transportation,apical blockage and ledge in NiTi groups.The operative time was shorter and endodontic flare-up seldom occurred in NiTi groups.Conclusion The NiTi rotary instrumentation technique could be used to prepare curved root canals effectively and quickly.The future use of NiTi engine-driven rotary in- strument appeared to be promising.

In this study, a combination of metabolomics and network pharmacology was used to study the pharmacodynamic substances and mechanism of action of Yiyi Fuzi powder (YYFZ) on rheumatoid arthritis (RA) rats. The animal experiments were conducted in accordance with the requirements of the Experimental Animal Ethics Committee of Tianjin University of Traditional Chinese Medicine (approval number: TCM-LAEC2021241). The metabolomic analysis using UPLC-Q-TOF/MS technique identified 22 metabolites, including arachidonic acid, tryptophan, linoleic acid, phenylalanine, as significant biomarkers for the treatment of RA with YYFZ, and they were significantly regressed after YYFZ treatment. The analysis of YYFZ blood components also revealed that 11 blood components, including hypaconitine, benzoylhypaconitine, and deoxyaconitine, may be the components that exert direct pharmacological effects in YYFZ in vivo, and further network pharmacological analysis of blood components obtained that YYFZ may exhibit anti-inflammatory effects through acting on PI3K/Akt signaling pathway, estrogen signaling pathway, vascular endothelial growth factor (VEGF) signaling pathway. The results of this study provide implications for the clinical application of YYFZ.

To reveal the possible mechanism of silica deposition in higher plants, lignin was isolated from rice straw following a modified method to conduct a simulation experiment in vitro. UV and infrared absorption spectra showed that the substance had the unique characteristics of pure lignin. The presence of silicon in the precipitation was revealed by TEM (transmission electron microscopy) with EDXA (energy dispersive X-ray analysis) device. It was found that in the borax solution where lignin precipitation occurred silica-lignin co-precipitation was produced but not in the DMSO solution where lignin was broken into its composition compounds and did not precipitate. This means that it is macromolecular lignin itself but not its compounds that could induce silica deposition in higher plants.
Lignin ; analysis ; chemistry ; metabolism ; Microscopy, Electron, Transmission ; Oryza ; chemistry ; metabolism ; Silicon Dioxide ; analysis ; chemistry ; metabolism ; Spectrophotometry, Infrared ; Spectrophotometry, Ultraviolet

OBJECTIVETo compare the effect of massage alone or in combination with pricking collateral blood therapy (PCBT) on mild carpal tunnel syndrome (CTS).

METHODSSixty cases with mild CTS were randomly assigned into two groups, the control group treated with massage therapy alone, and the treated group with PCBT at the six Jing points of the affected upper limb combined with massage, 30 cases in each group. The therapy was conducted in all patients for 3 courses. The therapeutic effect was observed and the differential value (D-value) of sensory conduction velocity (SCV) between the median and ulnar nerves of the fourth digit (D4) was detected before and after each course of therapy.

RESULTSAfter 3 treatment courses, excellent, therapeutic effect was shown in 26 patients and good effect in 4 of the treated group, while in the control group, the effect was excellent in 8, good in 20, and improved in 2. The effect was better in the former group than that in the latter (P<0.05). Marked difference in the D-value of SCV was shown at the end of each course in both groups (P<0.05). As compared it between the two groups, significant difference was shown at the end of the 1st and 3rd course respectively (P<0.05).

CONCLUSIONCombined treatment of massage and PCBT showed better effect on mild CTS than massage therapy alone, and the onset time may positively correlated to the frequency of applying PCBT.

Acupuncture Therapy ; methods ; Adult ; Carpal Tunnel Syndrome ; therapy ; Combined Modality Therapy ; Female ; Humans ; Male ; Massage ; Middle Aged

Objective:To investigate the proliferation of rat glomerular mesangial cells (GMC) influenced by different ethanol extracts of Tripterygium wilfordii HooK F.(TWHF).Methods:An HPLC method was used to establish the fingerprints of 5 different ethanol extracts of TWHF,and GMC was chosen to study the effects of different ethanol extracts of TWHF on cell proliferation.After statistical analysis,the spectrum-activity relationship was analyzed by using partial least squares regression(PLSR).Results:The HPLC fingerprints of the 5 different ethanol extracts of TWHF were established,and 32 characteristic peaks were characterized by the HPLC fingerprints.60%,70% and 95% ethanol extracts and glycosides tablets showed dose-effect relationship,and with the increase of dose,the more significant inhibition of cell proliferation was exhibited.The absorbance values of the 60% ethanol extracts at medium and high doses were lower than those of the other extracts at the same dose.The proliferation inhibition rate of GMC was used as the potency index and analyzed by PLSR,and 20 peaks were potency peaks at high dose(40 μg·L-1),17 ones were potency peaks at medium dose(20 μg·L-1) and 15 ones were potency peaks at low dose(10 μg·L-1).Conclusion:Part of the potency peaks has regular dose-effect relationship with the changes of dose.

Objective To investigate the echocardiographic features and clinical significance of prenatal diagnosis of total anomalous pulmonary venous connection (TAPVC). Methods Fetal echocardiographic images of 13 fetuses with TAPVC conifrmed by pathology or postnatal echocardiography were reviewed. Echocardiographic features and clinical signiifcance of prenatal diagnosis of TAPVC were summarized. Results Twelve fetuses with TAPVC were diagnosed prenatally by fetal echocardiography, including seven cases of supracardiac type, three cases of infracardiac type and two cases of intracardiac type. The common echocardiographic characteristics of 12 fetuses with TAPVC included slightly size discrepancy of left heart and right heart, large foramen ovale with increased shunting at the atrial level, increased distance between left atrium (LA) and descending aorta, absent insertions of pulmonary veins in the LA, presence of pulmonary venous conlfuence on the top of LA and dilatation of vessels where pulmonary venous conlfuence drained. One case was missed prenatally and intracardiac type TAPVC was diagnosed by postnatal echocardiography. Among the 13 cases, three were isolated and the other ten were all in association with other abnormalities. Conclusions There are fetal echocardiographic characteristics of TAPVC. Fetal echocardiography plays an important role in prenatal diagnosis of TAPVC.

OBJECTIVE Transient outward potassium current (Ito) plays a crucial role in cardiac phase 1 repolarization and the channels are assembled by pore-forming α-subunits (Kv4.2 or Kv4.3) and auxiliary subunits (KChIP2 and DPP6). Previous studies have found that the compound NS5806 increases Ito in canine ventricular cardiomyocytes through slowing current decay. Here, we reported that NS5806 produced an acute inhibitory action on Ito in mouse ventricular cardiomyocytes and human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CM). METHODS Whole-cell patch-clamp was used to record Ito in native myocytes and in HEK cells expressing cloned Kv4.x/KChIP2/DPP6 channels; Western-blot detected the channel protein expression. RESULTS In isolated mouse ventricular cardiomyocytes, NS5806 (0.1-30 μmol·L-1) inhibited Ito in a concentration-dependent manner with IC50 of 6.6±1.9 μmol·L-1. The current decay was significantly accelerated with a time constant from 53.8±5.5 to 41.8±3.0 ms at +60 mV (P<0.01). Similarly, NS5806 concentration-dependently reduced the Ito peak current amplitude with an acceleration of current decay. In addition, NS5806 increased IKv4.2/KChIP2 and delayed current decay, but decreased IKv4.2/KChIP2/DPP6 with the acceleration of current decay. The inhibitory action on the current was more potent if DPP6 expression level was increased from Kv4.2/KChIP2/DPP61:1:1 to 1:1:3. Western-blot showed a higher expression of DPP6 protein in mouse heart and in hiPS- CM compared to canine heart. Moreover, specific knock- down DPP6 expression by siRNA antagonized the inhibitory action of NS5806 in hiPS-CM. Our results pointed to an important role of DPP6 subunit in the regulation of NS5806 on the channel. By using molecular docking simulation, five interaction sites with high possibility between KChIP2 and DPP6 were identified. Mutations of those sites changed the inhibitory action of NS58056 into excitatory effect on the current with the delay of current decay. CONCLUSION NS5806 significantly inhibits Ito by accelerating current decay in mouse cardiomyocytes and hiPS-CM. The effect depends on the interaction between DPP6 and KChIP2 subunits.