The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Glucuronidase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Scutellaria baicalensis ; chemistry ; enzymology ; genetics ; Sequence Alignment

AIM: To establish HPLC fingerprint to make the difference between pre-and post processed Radix et Rhizoma Gentiana rigescen with wine-fried method. METHODS: Chromatographic fingerprint between pre-and post processed Radix et Rhizoma Gentiana rigescen with wine-fried was investigated by RP-HPLC and the gradient elution with acetonitrile and 0.1% phosphoric acid was applied to chromatographic separation(Alltima C_18(4.6 mm ?250 mm,5 ?m) column) for 65 min,and detection wavelength of 225 nm.Data were analysed by fingerprint similarity evaluation software to compare the similarity among the samples. RESULTS: The similitude degree of the10 batches of pre-and post processed Radix et Rhizoma Gentiana rigescen with wine-fried method were between 0.935-0.997 and 0.964-0.999,respectively.Comparing with the common processing of the HPLC fingerprint of Radix et Rhizoma Gentiana rigescen,the similarity of Radix et Rhizoma Gentiana rigescen processed with wine-fried was greater than 0.900. CONCLUSION: There have no significant deviation between semipolarity components of the pre-and post processed Radix et Rhizoma Gentiana rigescen with wine-fried method. The proportion of the main relative peak area has changed.

Among various ECL systems,such as 9,10-diphenylanthraeene,lucigenin,tris(2,2'-bipyridyl) ruthenium,peroxyoxalate,luminol,graphene,and nanocrystals,Ru(bpy)_3~(2+) ECL is one of the most widely studied ECL systems in recent years due to its broad applications in immunoassays,DNA probe assays,core-actants analysis,and aptasensors. In this review,the progress in Ru(bpy)_3~(2+) ECL has been summarized on the whole,and the future research trends have been proposed.

Objective To evaluate emergency nursing methods to permanent pacemaker patients with severe complication and acute hemodynamic changes during implantation. Methods The emergency nursing methods were analyzed retrospectively in 9 of 2 027 patients over 6 years, who had serious complication occurred during the pacemaker implantation Results There were 4 patients with pneumothorax, 2 with pericardial tamponed, 1 with ventricular fibrillation and 2 with acute left ventricular failure in 9 patients (0.4%). Observed the changes of symptoms and signs carefully,found the severe complications at the first time and performed the specific nursing based on different kinds of conditions and improved the prognosis of these patients. Conclusions The serious complications during the implantation of permanent pacemaker were life-threatening, and the specific nursing scheme to different types of complications could increase the successful rate of emergency service.

Objective To investigate the effects of Inula Britannica on myocardial caspase-3 and cytochrome c ( cyt c) following overtraining-induced acute myocardial injury in rats.Methods Forty-eight male Wistar rats weighing 200-220 g were randomly divided into 3 groups:group control (group C,n =8) ; group exhausting swim (group E,n =24) and group Inula Britannica (group IB,n =16).The animal model of overtraining-indnced acute myocardial injury was developed by exhausting swim.The animals were forced to swim until they were exhausted.The animals sank to the bottom and no righting reflex or escape response was elicited when they were taken out of water in groups E and IB.In group IB oral Inula Britannica 25 ml/kg was given 24 h and immediately before overtraining.Blood samples were taken from inferior vena cava immediately and at 6,24 h after overtraining in group E and at 6,24 h after overtraining in group IB for determination of serum cardiac troponin I (cTnI) concentration (by ELISA).The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and determination of caspase-3 and cyt c expression (by immuno-histochemistry).Results Overtraining significantly increased serum cTnI concentration and up-regulated myocardial caspase-3 and cyt c expression in group E as compared with group C.Oral Inula Britannica significantly attenuated overtraining-induced increase in serum cTnI concentration and myocardial caspase-3 and cyt c expression in group IB as compared with group E.Conclusion Inula Britannica can reduce overtraining-induced acute myocardial injury by down-regulating caspase-3 and cyt c expression.

Objective:To evaluate the heart hemodynamics in fetuses with premature ductus arteriosus constriction or closure using fetal heart quantification (FHQ).Methods:The clinical data of 50 singleton fetuses with ductus arteriosus constriction ( n=35) or ductus arteriosus closure ( n=15) who underwent echocardiography in Department of Ultrasound, Beijing Anzhen Hospital were retrospectively analyzed, from May 2013 to January 2020. Fifty healthy singleton fetuses were randomly selected as the control group. The ductus arteriosus diameter (DA), pulsatility index (PI), diameter of the left atrium(LA) and right atrium(RA), diameter of the left ventricle (LV) and right ventricle (RV), tricuspid regurgitation/right atrium area ratio (TR/RA Ratio), pressure gradient of tricuspid regurgitation (PG of TR), and heart/chest ratio were measured using conventional fetal echocardiography; the correlations among the parameters were analyzed. Speckle-tracking analysis was used to analysis and compute the LV and RV global spherical index (GSI), fractional area change (FAC) and global strain (GS), the LV ejection fraction(EF) and stroke volume (SV). These variables and their correlations were compared and analyzed. Results:Compared with the control group, the GS and FAC of the LV and RV in the ductus arteriosus constriction or closure groups were lower ( P<0.05) while the LV-SV was higher ( P<0.05). The FAC, GS, and EF values of the LV were higher in the premature ductus arteriosus closure group than in the ductus arteriosus constriction group ( P<0.05), while the RV-FAC was lower ( P<0.05), the RV-GS and LV-SV showed no significant changes ( P>0.05). Correlation analyse showed that the PI was positively correlated with DA( r=0.364, P<0.05); the PG of TR was negatively correlated with DA( r=-0.414, P<0.05); the TR/RA Ratio was negatively linearly correlated with PI( r=-0.388, P<0.05), and positively correlated with RV/LV Ratio ( r=0.369, P<0.05); the other parameters were not significantly correlated with the DA or PI ( P>0.05). Conclusions:Fetal heart hemodynamics in the premature ductus arteriosus constriction or closure groups change significantly, FHQ can provide valuable information for the evaluation of the fetal heart with ductus arteriosus constriction or closure.

Blepharospasm is a focal dystonia of the orbicularis oculi muscles, producing excessive eye closure. The etiology and pathogenesis is still unclear now. lt is usually appearing in adult period and predominant in females. The symptoms are typically triggered by stress, fatigue, intense light or individual factors. At advanced stages patients develop functional blindness. At present the main treatments include: botulinum toxin ( BTX ) , surgical procedures, systemic and ocular drugs and traditional Chinese medicine treatment. BTX administration has been an effective treatment. Surgical procedures have good effect but should be limited to the rare patients that do not respond to botulinum toxin treatment. A great variety of drugs have poor results. Chinese medicine has a certain therapeutic effect. Transcranial magnetic stimulation can improve symptoms. The epidemiology, anatomy, physiology, clinical manifestations, differential diagnosis, pathogenesis and treatment system were reviewed in this paper.

To observe the effect of various doses of oil of Piper longum unsaponifiable matter (OPUM) to cholesterol gallstones in experimental mice. C57BL/6 mice (n = 60) were randomly divided into 6 groups: control group, model group, OPUM (15, 30 and 60 mg x kg(-1)) group and ursodeoxycholic acid (UDCA, 60 mg x kg(-1)) group, administered for 10 weeks. The level of serum lipid and liver function enzymes were tested. The gallbladder was removed and bile was obtained by centrifugation. Next, the levels of the bile total cholesterol (TC), phospholipid (PL) and bile acid (TBA) were measured. The indicators of lipid peroxidation were determined and cholesterol saturation index (CSI) was calculated. The liver histological changes were observed by HE staining. The results showed that serum TC, TG (triglycerides) and AST (aspartate transaminase) contents, gallbladder cholesterol crystallization and CSI increased significantly (P < 0.05). In addition, the activity of SOD decreased significantly and MDA content increased significantly in liver (P < 0.05). HE staining results showed that the hepatic cord disorder and intracellular lipid droplets increased significantly. All results indicate that lithogenic diet lead to the formation of cholesterol gallstones. In OPUM (30 and 60 mg x kg(-1)) group, serum TC, TG and AST content, gallbladder cholesterol crystallization and CSI decreased significantly, the activity of SOD increased significantly and MDA content decreased significantly. HE staining results showed that OPUM can improve the morphology of liver cell, reduce the degree of hepatic cord disorders and restore the cell morphology close to normal. The cause of OPUM prevents cholesterol gallstone formation maybe due to protect the integrity of the liver cells, lower CSI, and reduce cholesterol crystal formation and hence prevent cholesterol gallstone formation.

AlM: To investigate the relationship between the subfoveal choroidal thickness ( SFCT) and both choroidal hemodynamic index and glycosylated hemoglobin in diabetic subjects.METHODS:Seventy-eight type 2 diabetic patients (156 eyes) from ophthalmology and endocrinology ward of our hospital were enrolled in this study, including 39 females and 39 males, with a mean age of (59. 8±6. 2)years. According to early treatment diabetic retinopathy study ( ETDRS) grading method, all samples were divided into diabetic retinopathy ( DR ) group, mild or moderate nonproliferative diabetic retinopathy group, severe nonproliferative diabetic retinopathy ( NPDR) group and proliferative diabetic retinopathy ( PDR ) group. The SFCT and choroidal hemodynamic index were measured by enhanced depth imaging optical coherence tomography ( EDl-OCT ) and Color Doppler lmaging. Recording glycosylated hemoglobin content of all samples. Using multivariate linear regression to analyse the relationship between the SFCT and both choroidal hemodynamic index and glycosylated hemoglobin.RESULTS: The end diastolic velocity ( EDV ) was significant higher and the SFCT was significant thinner in no diabetic retinopathy ( NDR) group than other groups. There was no significant difference of peak systolic velocity ( PSV ) between four groups. The resistance index ( Rl) was significant higher in severe NPDR group than NDR group and mild or moderate group, the Rl in PDR group was hihgest than other group with statistically significance. The SFCT was correlated positively ( b =0. 540,P<0. 001) with the glycosylated hemoglobin. No significant correlation was found between the SFCT and the choroidal hemodynamic index (DR,P=0. 341;PSV,P=0. 770;EDV,P=0. 131;Rl,P=0. 084).CONCLUSlON: Our results suggest that there is no significant correlations between the SFCT and the choroidal hemodynamic index; glycosylated hemoglobin is one of the factors that affect the SFCT in diabetic patients.

OBJECTIVETo investigate the therapeutic effect of IL-6 mAb on experimental autoimmune myocarditis (EAM) in rats, and search the mechanism of the role of IL-6, helper T cells 17 (Th17) and regulative T cells (Treg) in EAM pathogenesis.

METHODSThirty-four Lewis rats were divided into three groups randomly, i.e. control group (n = 6), EAM group (n = 12), and IL-6 mAb intervention group (n = 16). Rats in EAM group and IL-6 mAb intervention group were injected intracutaneously with myosin to establish EAM model. Rats in IL-6 mAb intervention group were injected intraperitoneally with 1 mg IL-6 mAb on 1st, 7th to 20th day after cardiac myosin immune injection. Myocardial inflammation was examined by HE stain, Masson stain, and TdT assay (TUNEL reaction) on 21st and 84th day after IL-6 mAb therapy in order to assess the therapeutic role. Spleen cells were analyzed by flow cytometry to illustrate Th17 and Treg cells? number and function. The serum concentration of IL-6, IL-10, IL-17, and TGF-beta in each group was measured by ELISA, concentration of STAT3, RORgammat, and Foxp3 mRNA in each group was determined with RT-PCR. Spleen cells derived from EAM were stimulated by IL-6 mAb in vitro, and the concentration of IL-10, IL-17 and TGF-beta was measured by ELISA.

RESULTSInflammation score, fibrosis score, and apoptosis index in IL-6 mAb intervention group were significantly decreased as compared with those in EAM group (P < 0.01). The number of Th17 and Treg cells in EAM group on the 21st day (experimental acute peak stage) were increased, and those in intervention group on the 21st day were significantly inhibited (P < 0.01). The concentration of serum IL-6, IL-10, IL-17 and TGF-beta in intervention group on the 21st day was decreased dramatically in comparison with that in EAM group on the same day (P < 0.01). The levels of peripheral blood STAT3, RORgammat, Foxp3 mRNA in intervention group on the 21st day was decreased significantly as compared with that in EAM group (P < 0.01). The expression of IL-10, IL-17 and TGF-beta was increased significantly (P < 0.01) by stimulation of IL-6 mAb on spleen cells derived from EAM in vitro.

CONCLUSIONSIL-6 mAb could neutralize IL-6, and ameliorate myocarditis and reduce heart autoimmune responses. IL-6 mAb has significantly protective effects on EAM by suppressing Th17 and Treg cells.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Autoimmune Diseases ; drug therapy ; immunology ; Disease Models, Animal ; Forkhead Transcription Factors ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-17 ; metabolism ; Interleukin-6 ; immunology ; Male ; Myocarditis ; drug therapy ; immunology ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Rats ; Rats, Inbred Lew ; STAT3 Transcription Factor ; metabolism ; Th17 Cells ; immunology ; Transforming Growth Factor beta1 ; metabolism