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.

Agricultural Workers' Diseases ; Humans ; Musculoskeletal Diseases

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.

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

OBJECTIVETo investigate the expression of microRNA-10a-5p (miR-10a) in laryngeal epithelial premalignant lesions (LEPL) and to analyze the correlations of its dysregulation with clinicopathologic parameters of LEPL.

METHODSAccording to the WHO classification (2005), 94 cases of LEPL were grouped into mild dysplasia (MID), moderate dysplasia (MOD), severe dysplasia (SD) and carcinoma in situ ( CIS). The expression of miR-10a in 94 cases of LEPL was detected by quantitative real-time polymerase chain reaction (qRT-PCR), and correlated with the clinical and follow-up data of all LEPL patients.

RESULTSmiR-10a was down-regulated in LEPL with increasing grade of dysplasia. There was significantly statistical difference between low-risk ( MID + MOD) and high-risk ( SD + CIS) lesion groups (P = 0.03). The linear regression analysis showed that miR-10a was correlated with grade and gender of LEPL (P < 0.05).

CONCLUSIONSDown regulation of miR-10a may be a useful molecular marker for grading of LEPL and diagnosis of early laryngeal cancer.

Biomarkers ; Carcinoma in Situ ; metabolism ; pathology ; Down-Regulation ; Humans ; Hyperplasia ; pathology ; Laryngeal Neoplasms ; metabolism ; pathology ; Larynx ; pathology ; MicroRNAs ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; RNA, Neoplasm ; metabolism ; Real-Time Polymerase Chain Reaction

Objective:To construct double copy and x gene deleted HBV expression plasmid and study its expression in Hep3B cell line. Methods:The double copy HBV DNA ( adr Ⅰ) was used to inactivate HBV x gene by inserting mutation and gene recombination. The inserted 55 bp DNA sequence was synthesized artificially; the insertion point was ApaL Ⅰ of x gene area. After recombination, an x gene defected HBV plasmid containing single P, S and C gene was constructed,which can express in mammalian cell line. Another plasmid carrying double copy HBV DNA with normal x gene was constructed as contrast. Both were used to transfect Hep3B cells. Then the cells were screened by G418 and HBV virus in culture medium were isolated and detected by fluorescence quantitative PCR. Results: Plasmids pcDNA3 KN F1F2 and pcDNA3 ES HBV2 were constructed successfully. After cell transfection, the HBV DNA was highly expressed with both plasmids on the 3 rd, 6 th,14th day. Conclusion: The plasmids constructed can express in Hep3B cell line and cause HBV replication; x gene defected HBV gene has no effect on HBV replication in Hep3B cell line.

OBJECTIVE: To study whether negative-pressure septal drainage could be an alternative to packs after septoplasty. METHOD: This was a randomized controlled trial. The study involved 60 patients who underwent septoplasty. Patients were randomly divided into two groups, one with anterior nasal packs and the other with negative-pressure septal drainage. Patients were asked to record pain levels using a visual analogue scale (VAS). Postoperative symptoms and complications were compared during 24 h and 48 h postoperative period including pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage. VAS scores and incidence were evaluated during 1 week and 6 weeks postoperative period including pain, bleeding, haematoma, septal perforation, synechiae and septal perforation. RESULT: Patients of negative-pressure septal drainage suffered from less pain than patients of nasal packs during the first 24 h and 48 h postoperative period. The results for pain, drying sensation of mouth, sleep difficulty, conjunctival congestion, haemorrhage were different between groups (P < 0.05), especially the amount of bleeding during 48 h postoperatively in patients undergoing negative pressure drainage [(0.52 ± 0.63)ml] was significantly less than the group who received anterior nasal packs [(21.03 ± 5.88) ml] (P < 0.01). On the other hand, haematoma, synechiae and perforation were not statistically different between groups during 1 week and 6 weeks follow-up period (P > 0.05). CONCLUSION Using negative-pressure drainage instead of nasal packs after septoplasty seems a more reasonable option. The negative-pressure drainage technique may be the preferred option to provide higher patient satisfaction and has the same level of postoperative complica.tion to nasal packs as for septoplasty surgery.
Drainage ; Humans ; Nasal Septum ; surgery ; Nasal Surgical Procedures ; Negative-Pressure Wound Therapy ; methods ; Nose ; Pain Measurement ; Patient Satisfaction ; Postoperative Period ; Tampons, Surgical

AIM:To observe the influence of Lycium barbarum polysaccharide (LBP) on the PI3K/Akt/eNOS signaling pathways in ovariectomized rat myocardium .METHODS:Female SD rats (n=30) were divided into sham oper-ation group , ovariectomized group , progynova group , high-dose LBP group and low-dose LBP group .The serum levels of estradiol, lactate dehydrogenase (LDH) and creatine kinase (CK) were measured by ELISA.The myocardial contents of H2 S and oxidative stress injury-related indicators were also detected .The morphological changes of the myocardium were observed with HE staining.The expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium was deter-mined by Western blot .RESULTS: Compared with sham operation group , the serum level of estradiol , the content of H2 S, the activity of GSH-Px, and the expression of eNOS and PI3K/Akt pathway-related proteins in the myocardium in ovariectomized group were all decreased , and the levels of ROS and MDA in the myocardium were increased (P<0.05). The serum levels of LDH and CK were also increased .The arrangement of the myocardial cells was disordered , and the in-tercellular space was also increased in the ovariectomized group .Compared with ovariectomized group , the serum level of estradiol, the myocardial levels of H2S and GSH-Px, and the protein levels of eNOS and phosphorylated Akt were all in-creased in high dose group, while the levels of ROS and MDA in the myocardium were decreased (P<0.05).The serum levels of LDH and CK were also decreased .The morphological changes of the rat myocardium were improved in high dose group.CONCLUSION: LBP prevents and treats postmenopausal cardiovascular lesions through regulating PI 3K/Akt/eNOS signaling pathways in ovariectomized rats .