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

Agricultural Workers' Diseases ; Humans ; Musculoskeletal Diseases

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.

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.

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.

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 To assess the left atrial early function in pilots with essential hypertension by strian rate imaging (SRI). Methods Twenty-five hypertensive pilots without left ventricular hypertrophy and expansion (LVN) were selected. Twenty-ifve healthy pilots were included as control group. At the left ventricular apical four-chamber view, three-chamber view, two-chamber view, the systolic, early and late diastolic peak strain rate (SRs, SRe, SRa) curves were acquired by SRI in the basal section, the middle section and the upper section of the wall of left atrium. The average strain rate (mSRs, mSRe, mSRa) was calculated. The LA maximal, minimal and pre-systolic volume (LAVmax , LAVmin and LAVp), LA passive ejection fraction (LAPEF), LA active ejection fraction (LAAEF) , left atrial stroke volume (LASV), E/A and E/E′ were calculated. Results The LAPEF, LAAEF, LASV, E/A, E/E′were (38.96±6.43)%, (61.11±5.86)%, (14.74±5.33)mm, 1.23±0.08, 8.63±1.77 in the control group, and were (34.18±5.72)%, (63.23±6.89)%, (10.12±3.23)mm, 1.04±0.26, 7.16±0.84 in the study group. Compared with the control group, signiifcant difference was detect in LAPEF, LAAEF, LASV, E/E′(t=4.840, 2.690, 4.801, 2.950, all P<0.05). No signiifcant difference was found in E/A between the study and control group (t=-1.424, P=0.178). There was signiifcant positive correlation between SRa and E/E′(r=0.632, P<0.05). Conclusion SRI could be applied in the evaluation of early left atrial function. In pilots with essential hypertension, the changes of strain rate in left atrial wall, which is prior to the modeling geometry of left ventricular, could be detected by SRI.

Objective To detect the expression of chemokine receptors CXCR3 mRNA in the peripheral blood mononuclear cells (PBNCs) of patients with Rheumatoid Arthritis (RA) and to analyze the relationship between the expression and the disease activity. Methods mRNA was extracted from PBNCs and the expression of CXCR3 mRNA was detected by real-time fluorescence quantitative PCR (RFQ-PCR) in 51 RA patients and 32 controls. T-test, x2-test, ANOVA were used for statistial analysis. Results Comparison between the two groups had shown that the expression levels of CXCR3 mRNA in clinical active RA group were higher than those of the RA patients in remission and healthy controls (P<0.05). The expression levels of CXCR3 mRNA were positively correlated with serum levels of ESR and CRP in clinical active RA group (r=0.824, r=0.765, P<0.05). In addition, RF titer, APF, AKA, and anti-CCP had no significant correlation with the expression levels of CXCR3 mRNA in RA patients (P>0.05). Conclusion RFQ-PCR is a sensitive,reproducible and practical test. The mRNA expressions of CXCR3 are significantly elevated in RA patients,which suggest that CXCR3 may be involved in the pathogenesis of RA. The mRNA expressions of CXCR3 in active RA patients are higher than those of RA patients in remission. These results indicate that CXCR3 may play an important role in the pathogenesis and progression of RA, and CXCR3 may be considered as an indicator for disease activity, therapeutic efficacy and prognosis of RA.

OBJECTIVETo analyze the reliability and validity of the Fatigue Self-assessment Scale (FSAS).

METHODSThe scale was applied among the participants assigned to 4 groups, the differences in types, degrees and characteristics of fatigue of them were compared, and the reliability and constitutional validity of ESAS were assessed by internal consistency analysis, exploratory factor analysis and confirmatory factor analysis using the statistical software of SPSS and LISREL.

RESULTSStatistical differences of types, degrees and characteristics of fatigue presented in the participants of the 4 groups. The Cronbach's alpha of various factors in the scale were 0.772-0.908; the indexes for the section of assessing type, and degree of fatigue were RMSEA=0.065, NNFI=0.95, CFI=0.96; and those for the section of assessing characteristics of fatigue were: RMSEA=0.10, NNFI=0.93, CFI=0.96.

CONCLUSIONThe FSAS has good differentiability, reliability and constitutional validity for assessing the type, degree and characteristics of fatigue in various populations. In order to explore the relationship of TCM syndrome patterns with the type, degree and characteristics of fatigue, its future application for evaluation of fatigue and intervention effect of anti-fatigue should be combined with TCM syndrome differentiation.

Adolescent ; Adult ; Fatigue ; diagnosis ; epidemiology ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Models, Statistical ; Self-Examination ; Software ; Surveys and Questionnaires ; Young Adult

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