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

Objective To investigate the change of serum leptin and ghrelin levels in patients with active ulcerative colitis (UC) and their clinical significance.Methods Fifty-four patients with active UC and 25 normal controls were enrolled in this study,and serum levels of leptin and ghrelin were determined by enzyme-linked immunosorbent absorption.The correlations between serum leptin/ghrelin and the main clinicopathological features of patients with active UC were analyzed.The nutritional status of these patients were assessed by using the Nutritional Risk Screening 2002 ( NRS 2002).Results The serum levels of leptin and ghrelin in patients with active UC were significantly higher than those in normal controls [leptin:(245.51 ± 129.72) vs.( 183.80 ±59.67) pg/ml,P =0.029; ghrelin:(1477.96 ±501.86) vs.(1188.00±474.51) pg/ml,P=0.041]; and both of them were correlated with the disease severity ( leptin:r =0.715,P =0.019 ; ghrelin:r =0.667,P =0.027),but were not correlated with the clinical type,the extent of lesion,and the endoscopic grading (all P ＞ 0.05).In the UC patients with nutritional risk,the serum levels of leptin and ghrelin were also significantly higher than those in subjects without nutritional risk [ leptin:(332.82 ± 172.92) vs.( 194.15 ± 49.71 ) pg/ml,P =0.026 ; ghrelin: ( 1848.60 ±326.48) vs.(1259.93±459.24) pg/ml,P=0.021].Conclusion Patients with active UC have remarkably higher serum leptin and ghrelin levels,which is associated with the disease severity and the nutritional risk.

Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

To review the application,acceptance,evaluation and funding of the General,Youth and Regional Science Fund supported by Chinese National Natural Science Foundation in traditional Chinese medicine in 2017.There were 4 472 applications in the subject,114 of which failed in the formal examination,4 358 applications were accepted in the next assessment step;1 403 applications got more than 2 recommendations after peer assessment and 637 applications were funded in the end.The major problems in every step of Science Fund were analyzed in this paper.The author hopes all applicants should pay attention to the problems,and then improve the quality of applications in the field.

BACKGROUNDTo investigate the clinical features of hepatitis A in 1 629 children under 14 years of age treated in our department at various periods of time.

METHODSThe patients were divided into two groups: 1. Group A consisted of 883 patients treated from January 1984 to December 1990; 2. Group B consisted of 746 patients treated from January 1991 to December 2000. The clinical data of all the patients were retrospectively analyzed.

RESULTS1. The average age was 7.17+/-3.27 and 8.78+/- 3.28 years (chi2=0.54, P>0.05) and the mean course of disease 26.25+/-16.96 and 25.65+/-12.58 days (chi2=0.29, P>0.05). 2. Double peak or multi-peak serum ALT was found in 89 patients. Four peaks of serum ALT was found in one patient. 3. HBsAg was found positive in 143 patients (8.80%). The mean course of disease was 34.40+/-25.86 and 25.20+/-15.43 days (chi2=146.5, P<0.001) in HBsAg positive and negative patients, respectively. 4. Liver puncture biopsy in 26 patients with hepatitis A showed that there was piecemeal necrosis in 2 patients.

CONCLUSIONS1. There was no significant delay in age of children with HAV infection in 1990s. There was no marked difference in the course of disease between the patients simultaneously receiving various drugs and those receiving one or two drugs. 2. The double peak or multi-peak of serum ALT in patients with hepatitis A might be related to liver damage caused by HAV and immune mechanism. 3. The major type of virus for combined infection in patients with hepatitis A is HBV. The course of disease was prolonged with combined infection of HBV. 4. Piecemeal necrosis might be seen in the liver of a small proportion of patients with hepatitis A alone, which may not be enough to suggest chronicity.

Adolescent ; Alanine Transaminase ; blood ; Child ; Child, Preschool ; Female ; Hepatitis A ; diagnosis ; therapy ; virology ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; blood ; Humans ; Infant ; Liver Function Tests ; Male ; Retrospective Studies ; Superinfection

Objective To study knowledge of healthy volunteers in-volved in Phase ⅠClinical Studies to clinical researches and related fac-tors.Methods The public awareness of research for therapeutic ad-vancements through knowledge and empowerment (PARTAKE) question-naires, designed by Duke University , were administered to 100 partici-pants in phaseⅠclinical study in Peking university third hospital.Re-sults The following perceptions were reported: research benefited our society (96.0%) , human experiment was necessary for developing new therapeutic method (91.0%), confidentiality and voluntary participating were adequately approved ( 90.0%) , all the undesirable effects were caused by the research ( 18.0%) , and volunteers would be paid for any undesirable adverse reactions completely ( 81.0%).Conclusion Re-sults suggest that the healthy volunteers have a good command of clinical research generally.They are aware of protections to participants , but they do not have a full understanding of compensation for damages .

Diabetes,a chronic metabolic disease with hyperglycemia,has a high incidence in the modern time that put many patient families into troubles. At present,insulin or oral hypoglycemic agents are used to treat diabetes,though several side effects such as hypoglycemia and various complications mainly caused by these drugs. Antibody with high selectivity and high therapeutic index is employed in many disease treatments. In this paper,the recent study of different antibodies against certain targets of T cells,B lymphocytes,glucagon receptors and vascular endothelial growth factor are involved,which could supply new insight into the treatment of diabetes.

Objective To evaluate left ventricular function in patients with hypertensive hypertrophic cardiomyopathy(HHC)using real-time 3-dimensional echocardiography(RT-3DE).Methods Thirty patients with HHC and 32 control subjects were studied.Full-volume RT-3DE data from apical window were acquired,and regional volumetric time curves of 17 segments were obtained by fast 3-dimensional border detection software.Several left ventricular function parameters were calculated semiautomatically,including global left ventricular end-diastolic volume(EDV),end-systolic volume(ESV),left ventricular ejection fraction(LVEF),the ratio of ESV/EDV of 17 segments,the standard deviation(SD)and difference(Dif)(adjusted by the R-R interval) of time to minimum systolic volume(Tmsv)in 16 segments(Tmsv16-SD and Tmsv16-Dif).Results EDV and ESV were significantly larger in patients with HHC than that in control subjects[(88±29)ml vs (72±15) ml,t=-2.680,P=0.008;(28±10)ml vs (22±6 )ml,t=-2.613,P=0.01].HHC had a higher ratio of ESV/EDV at interventricular septum(IVS)compared with control group[mid-segments of anterior IVS:(40.51±20.28)% vs (26.43±10.10)%,t=-3.378,P=0.002;mid-segments of posterior IVS:(41.44±23.55)% vs (24.46±8.12)%,t=-3.688,P=0.001;apical segments of IVS:(30.96±21.31)% vs (19.53±7.33)%,t=-2.745,P=0.01].In patients with HHC,Tmsv16-SD and Tmsv16-Dif were significantly longer[(2.48±1.38)% vs (1.16±0.26)%,t=-5.117,P＜0.001;(7.67±5.07)% vs (3.95±1.48)%,t=-3.865,P＜0.001].And the prevalence of left ventricular dyssynchrony was higher than that in control subjects(43% vs 3%).Conclusions HHC patients may have regional left ventricular systolic dysfunction before global changes,and have a higher prevalence of left ventricular dyssynchrony.RT-3DE is a useful imaging modality for assessing left ventricular systolic function.

Objective To evaluate the segmental myocardium of left ventricular wall in patients with myocardial hypertrophic cardiomyopathy (HCM) by TDI-Q, explore whether the segmental myocardium contractile function is changed or not and determine the myocardial mechanics parameters variation. Methods Thirty-two healthy volunteers and twenty-one patients with hypertrophic cardiomyopathy were included and the standard dynamic two-dimensional tissue Doppler imaging (TDI) of mitral, papillary muscle and apical short axis view were collected in three consecutive cardiac cycles. The mechanical parameters variation and characteristics of systolic radial peak displacement (RD) and time to peak in left ventricle subendocardial, mid-myocardium and epicardial myocardium at different level and segment were analyzed.Results In healthy control group, at left ventricular basal, apical and papillary muscle level, there was no significant difference for time to peak and systolic radial peak displacement (F=0.74, 1.28 and 1.79, all P>0.05). In patients with HCM, time to peak of systolic RD at left ventricular different level was asynchronous. Time to peak of RD in septum at papillary muscle levels and apical lateral wall were longer than those of other segments. In healthy control group, except for apical inferior and lateral wall, RD of subendocardial myocardium was significantly greater than that of epicardial myocardium at different segments (t=-1.903, 4.574,-3.552,-2.614,-1.728,-1.790,-1.836,-2.794 and 2.733, all P<0.05 ). In patients with HCM, RD of subendocardial myocardium was significantly greater than that of epicardial myocardium in posterior wall, septum at basal level and in inferior wall, posterior wall and lateral wall at papillary muscle level (t=-2.305,-2.148, 3.550,-1.182 and-3.602, all P < 0.05). At the same segment, transmural RD of subendocardial and subepicardial myocardium in healthy subjects were greater than that in patients with HCM. In inferior wall, posterior wall, lateral wall and septum at basal level, in inferior wall, posterior wall and septum at papillary muscle level, and in lateral wall and septum at apical level, differences of transmural RD were statistically significant (t=-3.787,-2.983,-4.325,-6.972,-2.352, 2.823,-3.274,-1.338 and-2.857, all P<0.05). Conclusions In patients with HCM, synchrony of left ventricular motion at different level was abnormal and transmural RD of endocardial and epicardial myocardium was decreased, which suggested regional systolic dysfunction. Ultrasound assessment of left ventricular segmental transmural mechanics can further reveal left ventricular mechanical characteristics in patients with hypertrophic cardiomyopathy.