Objective: To investigate the enzymatic hydrolysis of gypenosides by β-glucanase 26130 CN to prepare some hydrolyzed secondary saponins, so as to provide material basis for further biological studies. Methods: Using β-glucanase 26130 CN, the total saponins from Herba Gynostemmatis were hydrolyzed with the enzyme catalysis, and the hydrolytic products were analyzed by ultra high- performance liquid chromatography coupled with quadrupole time- of- flight mass spectrometry(UHPLC- Q- TOF/MSE)to identify the converted products. Then, the main components of Herba Gynostemmatis, gypenosides XLIX and A, were used as substrates of the β-glucanase 26130 CN for convertsion to secondary saponin products. The products were separated by preparative highperformance liquid chromatography(HPLC)and identified by NMR and MS. Results: Twenty eight triterpenoid saponins were identified in the total saponin hydrolysate on the basis of their high-resolution MS data, by comparison with the data in the literature, and seven of them were validated to be the converted products. It was found that the β-glucanase 26130 CN could hydrolyze the glycosidic bond of terminal glucose or xylose in the molecule of gypenosides. By the enzymatic hydrolysis of gypenoside XLIX and gypenoside A, gypenoside I(the one glucosyl-lost gypenoside XLIX)and gypenoside UL1(the one xylosyl-lost gypenoside A)were obtained via the preparative HPLC separation of the gypenoside XLIX and gypenoside A hydrolysates, respectively. Conclusion: β-glucanase 26130 CN could effectively catalyze the hydrolysis of terminal glucosyl and xylosyl groups in gypenosides, with a relatively high hydrolytic conversion rate, which could be used to prepare some secondary saponins or aglycones.

Objective:To investigate the effect of methylprednisolone that decrease plasminogen activator inhibitor-1(PAI-1) in plasma through mediating the PA/plasmin system.Methods:Employed immunohistochemical technique-ELISA to determine PAI-1 level in plasma.Results:①PAI-1 in LN patients is significantly higher than that in the control group (P0.05).Conclusion:It is thus proved that methylprednisolone is capable of decreasing PAI-1 in plasma through mediating the PA/plasmin system and two shocks is the minimum for effective cure. [

Objective: To study the mechanism of differentiation of rat ectomesenchymal cells to odontoblasts. Methods: Ectomesenchymal cells were cultured in three-dimension culture model using collagen gel as frame, and the change of phenotype of ectomesenchymal cells were observed and detected by phase-contrast microscopy and immunohistochemistry after the cells had been treated by 10 ng/ml of bFGF or/and 100 ng/ml IGF-1. Results: 4 days after treatment by bFGF and IGF-1, the cells appeared to be odontoblast-like cells aligned parallelly and polarized with long cytoplasmic processes attached to one end of the cell body.The cells were positive for DSP expression. However, the cells were DSP negative and aligned disorderly in other groups. Conclusion: Ectomesenchymal cells can be induced to differentiate to odontoblast-like cells in three-dimension culture model with the treatment by bFGF and IGF-1.

Objective: To compare the concentration of IL-1? in the apical exudates of phoenix abscess of chronic periapical periodontitis and to examine the correlation of IL-I? concentration with clinical and radiographic findings of the involved teeth. Methods: 35 single-rooted teeth diagnosed as phoenix abscess and 35 as chronic periapical periodontitis were examined. The periapical signs and symptoms were recorded. Radiographs were taken and periapical radiolucent areas were calculated with the help of the AutoCAD software. The standard paper-point sampling method was used to collect and quantify the periapical exudates. IL-1? in the exudates was detected by enzyme-linked immunosorbent assay (ELISA). All statistical analyses were finished with SPSS 10.0 software. Results: The phoenix abscess group showed significantly lower concentration(5.65?2.76) ng/ml of IL-1? in the exudates and larger radiolucent areas(32.10?13.82) mm 2 on the X-ray films than the chronic periapical periodontitis group[(12.51?5.15) ng/ml and (6.51?3.56) mm 2 respectively] (P

BACKGROUNDWhile echocardiography has been a pivotal screening test in pulmonary arterial hypertension (PAH), the presence of structural cardiac defects may affect the ability to reliably predict pulmonary artery pressures (PAPs). This study sought to evaluate the accuracy of Doppler echocardiography (DE) for estimating PAPs in adult atrial septal defect (ASD) patients with PAH.

METHODSA prospective study was carried out to compare the echocardiographic assessment of PAP with the same pressures obtained by right heart catheterization (RHC) in adult ASD patients with PAH who underwent simultaneous DE and RHC. Bland-Altman analyses were performed to evaluate the agreement between DE and RHC measurements of PAPs.

RESULTSTwo hundred and fifty-seven patients were included in the study. A significant overestimation of the systolic pulmonary arterial pressure (sPAP) and mean pulmonary artery pressure (mPAP) was reported by echocardiography compared with those by catheterization ((81.8 ± 26.9) mmHg vs. (72.9 ± 26.9) mmHg, P < 0.01; (51.9 ± 16.4) mmHg vs. (41.4 ± 17.2) mmHg, P < 0.01, respectively). Twenty-one percent (55/257) of the patients had PAH when estimated by echocardiography whereas showed normal results in the subsequent catheterization test. Using Bland-Altman analytic methods, the bias for the echocardiographic assessment of the sPAP was 9.1 mmHg with 95% limits of agreement ranging from -24.4 to 42.6 mmHg. For mPAP measurement, the bias was 10.5 mmHg with 95% limits of agreement ranging from -12.4 to 33.4 mmHg. On multiple linear regression analysis, age, gender, body surface area, ASDs' diameter, PVR, diastolic blood pressure, and echocardiographic assessment of right atrial pressure (RAP) explained 68.8% of the total variability in the model (r(2) = 0.688, P < 0.01).

CONCLUSIONInaccuracy was frequently reported in Doppler echocardiographic assessment of the PAP in adult ASD patients with PAH and was often associated with age, gender, body surface area, ASDs' diameter, pulmonary vascular resistance, diastolic blood pressure and echocardiographic estimation of RAP.

Adolescent ; Adult ; Aged ; Echocardiography, Doppler ; methods ; Female ; Heart Septal Defects, Atrial ; diagnosis ; Humans ; Hypertension, Pulmonary ; diagnosis ; Male ; Middle Aged ; Prospective Studies ; Pulmonary Artery ; pathology ; Young Adult

OBJECTIVE: To investigate the inhibitory effects of novel phosphodiesterase 4 inhibitor ZL-n-91 to the proliferation of leukemia cells L1210 and K562. METHODS: CCK-8 method was used to detect the effect of ZL-n-91 to the proliferation of L1210 and K562 cells, and the proliferation rate, IC RESULTS: ZL-n-91 showed a significant inhibitory effect to the proliferation of leukemia cells L1210 and K562 in a dose-dependent manner (P<0.001). After treated by ZL-n-91, the leukemia cells L1210 and K562 in the S-phase in cell cycle decreased significantly compared with those in control group (P<0.01). The apoptosis of leukemia cells L1210 and K562 could be induced by ZL-n-91 (P<0.001), and the expression level of apoptosis related protein BAX significantly increased. In the animal experiment, the result showed that ZL-n-91 could significantly inhibit the growth of subcutaneously transplantation tumor (P<0.05). CONCLUSION The novel phosphodiesterase 4 inhibitor ZL-n-91 can effectively inhibit the proliferation of leukemia cells L1210 and K562, which has the potential of anti-leukemia drug development.
Animals ; Cell Proliferation ; Humans ; K562 Cells ; Leukemia ; Mice ; Mice, Nude ; Phosphodiesterase 4 Inhibitors/pharmacology*

BACKGROUNDPercutaneous balloon pulmonary valvuloplasty (PBPV) is the preferred therapy for pulmonary valve stenosis (PVS). This study retrospectively reviewed recent PBPV outcomes in infants with PVS. The aim of this study was to evaluate factors associated with immediate therapeutic outcomes and restenosis during medium-term follow-up.

METHODSThe study included 158 infants with PVS who underwent PBPV from January 2009 to July 2015. Demographic characteristics and patient records were reviewed, including detailed hospitalization parameters, hemodynamic data before and immediately after balloon dilation, cineangiograms, and echocardiograms before PBPV and at each follow-up. All procedures were performed by more than two experienced operators.

RESULTSImmediately after balloon dilation, the pressure gradient across the pulmonary valve decreased from 73.09 ± 21.89 mmHg (range: 43-151 mmHg) to 24.49 ± 17.00 mmHg (range: 3-92 mmHg; P < 0.001) and the right ventricular systolic pressure decreased from 95.34 ± 23.44 mmHg (range: 60-174 mmHg) to 52.07 ± 18.89 mmHg (range: 22-134 mmHg; P < 0.001). Residual transvalvular pressure gradients of 67.31 ± 15.19 mmHg (range: 50-92 mmHg) were found in 8.2% of patients, indicating poor therapeutic effects; 6.4% of patients had variable-staged restenosis at follow-up and 3.8% underwent reintervention by balloon dilation or surgical repairs. Further analysis demonstrated that the balloon/annulus ratio showed statistically significant differences (P < 0.05) among groups with different therapeutic effects and between the restenosis and no-stenosis groups. Binary logistic regression analysis further revealed that higher balloon/annulus ratio (odds ratio: 0.005, 95% confidence interval: 0-0.39) was an independent protective factor for restenosis. The rate of severe complications was 1.9%.

CONCLUSIONSPBPV is a definitive therapy for infants with PVS based on its effectiveness, feasibility, and safety. Restenosis upon medium-term follow-up is relatively rare.

BACKGROUNDDrug resistance to targeted therapies occurs in lung cancer, and resistance mechanisms related to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are continuously being discovered. We aimed to establish a novel method for highly parallel multiplexed detection of genetic mutations related to EGFR TKI-resistant lung cancer using Agena iPLEX chemistry and matrix-assisted laser desorption ionization time-of-flight analysis on the MassARRAY mass spectrometry platform.

METHODSA review of the literature revealed 60 mutation hotspots in seven target genes (EGFR, KRAS, PIK3CA, BRAF, ERBB2, NRAS, and BIM) that are closely related to EGFR TKI resistance to lung cancer. A total of 183 primers comprised 61 paired forward and reverse amplification primers, and 61 matched extension primers were designed using Assay Design Software. The detection method was established by analyzing nine cell lines, and by comparison with LungCarta™ kit in ten lung cancer specimens. EGFR, KRAS, and BIM genes in all cell lines and clinical samples were subjected to Sanger sequencing for confirming reproducibility.

RESULTSOur data showed that designed panel was a high-throughput and robust tool, allowing genotyping for sixty hotspots in the same run. Moreover, it made efficient use of patient diagnostic samples for a more accurate EGFR TKIs resistance analysis. The proposed method could accurately detect mutations in lung cancer cell lines and clinical specimens, consistent with those obtained by the LungCarta™ kit and Sanger sequencing. We also established a method for detection of large-fragment deletions based on single-base extension technology of MassARRAY platform.

CONCLUSIONSWe established an effective method for high-throughput detection of genetic mutations related to EGFR TKI resistance based on the MassARRAY platform, which could provide more accurate information for overcoming cancers with de novo or acquired resistance to EGFR-targeted therapies.

Administration, Inhalation ; Adult ; Carbolines ; administration & dosage ; therapeutic use ; Female ; Heart Defects, Congenital ; drug therapy ; Humans ; Hypertension, Pulmonary ; drug therapy ; Iloprost ; administration & dosage ; therapeutic use ; Male ; Tadalafil ; Young Adult

OBJECTIVETo investigate the mutation and background of SLC22A5 in 6 patients with primary carnitine deficiency (PCD) who only presented as cardiomyopathy.

METHODGenomic DNA were abstracted from the blood of the patients and their parents. Using high-throughput sequencing to determine the mutation site.Using Sanger method to confirm the mutated alleles in PCD patients and detect the corresponding sequences in their patients. Using SIFT and PolyPhen to predict the function of protein for detected missense mutations.

RESULTThree different mutations were identified, including 2 nonsense mutations (R254X and R289X), 1 missense mutation (C113Y), R254X was the most frequently seen mutation. Four patients had compound heterozygous mutations and 2 patients had homozygous mutations. Their parents were found to have heterozygous mutations in corresponding alleles.

CONCLUSIONR254X, R289X and C113Y might be associated with primary carnitine deficiency.

Adolescent ; Base Sequence ; Cardiomyopathies ; genetics ; Carnitine ; deficiency ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; Female ; Genotype ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Hyperammonemia ; genetics ; Infant ; Male ; Muscular Diseases ; genetics ; Mutation ; Organic Cation Transport Proteins ; genetics ; Pedigree ; Solute Carrier Family 22 Member 5