Objective To explore whether Helq deletion affect the pluripotency of stem cells. Methods Helq knockout embryonic stem cells were obtained by CRISPR?Cas9 gene editing technique. Results The results of immunoflu?orescence analysis showed that the expression of Oct4 and Nanog had no obvious difference to that of the control cells. The Helq-/ - embryonic stem cells could produce viable pups by tetraploid complementation, indicating that their pluripotency was not affected. Meanwhile, we found that day 2 epiblast?like cells also were obtained through differentiation of the Helq-/ - embryonic stem cells in vitro. Immunostaining and real?time PCR analysis showed that the gene expression of Helq-/ - epiblast cells were similar to the wild type cells. Conclusions Taken together, it is proved that the genomic in?stability caused by Helq deletion does not affect the pluripotency of pluripotent stem cells.

OBJECTIVE:To study the chemical constituents in antioxidant activity part of Semiaquilegia adoxoides. METH-ODS:The antioxidant activity part of S. adoxoides were isolated and purified by chromatography on silicagel and Sephadex LH-20 column,and compound structures were identified physicochemical properties and spectral data analysis. RESULTS:Nine com-pounds were isolated from the antioxidant activity part of S. adoxoides,namely magnoflorine (1),griffonilide (2),salidroside (3),ferulic acid(4),genistein(5),2,4-dihydroxybenzoic acid(6),chlorogenic acid(7),caffeic acid(8)and p-coumaric acid (9). CONCLUSIONS:The study confirms the active material basis in antioxidant activity part of S. adoxoides for the first time.

BACKGROUND:Studies have shown that the finite element method could preferably simulate the biomechanical analysis for the object with complicated structures and irregular shapes. The similarities for the finite element model have great influences on the results of the analysis. However, to construct an ideal model is the most time-consuming and complicated portion for the finite element analysis. OBJECTIVE:To construct a finite element model for the maxilary first molar and the periodontal tissue, and to provide a basis of biomechanical researches of the maxilary first molar. METHODS: A volunteer with complete mandibular dentition and healthy periodontal tissue was selected in this study. Cone-beam CT was scanned. The images were saved as DICOM format. These images were imported to the medical modeling software Mimics. The surface model for the maxilary first molar and the alveolar bone was constructed. The model was then imported to GiD for pre-processing. Thus, the complete three-dimensional finite element model for the maxilary first molar and the periodontal tissue was constructed. RESULTS AND CONCLUSION:A finite element model for bilateral maxilary first molar, periodontal ligament and maxilary alveolar bone was constructed, including 896 035 nodes and 4 881 067 elements. This model has restored the geometric shape and the structure of the research object. This study successfuly constructed finite element models of maxilary first molar and the periodontal tissue, which can be a basis of biomechanical researches for the maxilary first molar and the periodontal tissue under the effect of different clinical orthodontic forces.

Objective To investigate the apoptotic effect of microRNA-1 (miR-1) on hypoxemic cardiomyocytes. Methods The cultured H9C2 cells were divided into 5 groups:normal control group, negative control group, H2O2 group, miR-1 group and H2O2+miR-1 group. After verified the success of transfection by real time PCR, MTT and flow cytometry methods were used to test the cell vitality and apoptotic rate, while the mRNA and protein expression level of Bcl-2 were de-tected by real time PCR and Western blot methods. Results Compared with normal control group, there were no significant differences in all indexes in negative control group. The application of H2O2 and miR-1 respectively or together significantly increased the miR-1 level and apoptotic rate, and reduced the cell vitality and Bcl-2 expression level. Conclusion mi-croRNA-1 can induce cardiomyocyte apoptosis by downregulating anti-apoptosis factor Bcl-2.

Objective To explore the clinical significance of the brain natriuretic peptide(BNP) in evaluating the cardiotoxicity caused by daunorubicin(DNR) through studying the changes of the plasma BNP levels in children with acute leukemia who accepted the chemotherapy with DNR.Methods Thirty-one children with acute lymphoblastic leukemia(ALL) admitted in the year of 2002-2004 underwent the chemotherapy in DVLP project.The plasma level of BNP was measured by enzyme linked immunosorbent assay(ELISA) and the left ventricular end diastolic diameter(LVEDD) by color Doppler respectively before and after the administration of DNR.Simultaneously,electrocardiography(ECG) and cardiac muscle enzymes(LDH1,CK-MB) were measured as routine.Results The plasma level of BNP increased from(3.97?2.41) ng/L to(18.25?7.63) ng/L(P

The Ca²⁺-activated monovalent cation selective transient receptor potential melastatin 4 (TRPM4) channel expressed in pancreatic β-cells is implicated in the β-cell function and insulin secretion, but how pharmacological function of TRPM4 channel affects membrane excitability of β-cells remains largely unknown. Here, we report that pharmacological inhibition of TRPM4 by specific inhibitor 9-phenanthrol attenuates electrical activities of pancreatic β-cells. In whole-cell current clamp recordings, 9-phenanthrol results in inhibition of action potential frequency induced by tolbutamide of the INS-1 pancreatic β-cells in a dose-dependent manner with an IC₅₀ value of 14.99 ± 7.93 μmol·L^-1. Similarly, 9-phenanthrol also inhibited action potential firing in INS-1 cells stimulated by current injection. Further recordings of β-cells demonstrate the significant inhibitory effects on action potential peak and action potential amplitude by 9-phenanthrol. Taken together, our results show the involvement of TRPM4 channel function in pancreatic β-cells depolarization and action potential, it provides pharmacological experimental methods and theoretical support for the study of TRPM4 channel in pancreatic β-cells.

OBJECTIVETo investigate the effect of Zige lyophilized powder for injection in improving the acute cerebral microcirculation disturbance in rats.

METHODWindow craniotomy was performed for rats after the drug administration for 14 days. The experimental microcirculation disturbance model was duplicated with high molecule dextran. After the drug administration, the micro-vein diameters of cerebral pla mater of various groups were observed and recorded under the biological microscope. The blood flow volume was monitored by laser Doppler flow-meter. HCT was measured by the electric resistance method. The hemorheological indexes were detected by the auto-hemorheological instrument.

RESULTZige lyophilized powder for injection (16.40, 32.70, 65.40 mg x kg(-1)) could significantly expand the micro-vein diameter of cerebral pla mater, improve the downward trend of the blood flow volume, and reduce the various hemorheological indexes.

CONCLUSIONZige lyophilized powder for injection shows the effect in improving the cerebral microcirculation.

Animals ; Brain ; blood supply ; drug effects ; Brain Ischemia ; drug therapy ; physiopathology ; Cerebrovascular Circulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Powders ; administration & dosage ; Rats ; Rats, Sprague-Dawley

A fast and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of prodrug of paclitaxel (Pro-PTX) and paclitaxel (PTX) in rat plasma was developed. The plasma samples were subjected to protein precipitation with acetonitrile (0.1% formic acid), and then separated by LC with an Ultimate AQ-C18 column (50 mm × 3.0 mm, 3 μm) and acetonitrile-1 mmol·L^-1 ammonium formate (containing 0.1% formic acid) as the mobile phase. Multiple reaction monitoring (MRM) scanning mode was used to detect the ion responses m/z 983.4→415.2 (Pro-PTX), m/z 854.4→286.1 (PTX) and m/z 808.3→527.2 (Docetaxel, internal standard) by using a triple quadrupole tandem mass spectrometer with electrospray ionization source and positive ion mode. The method validation results show that the linear ranges of Pro-PTX and PTX in plasma were 2.00-500 ng·mL^-1 (r > 0.99) and 4.00-1 000 ng·mL^-1 (r > 0.99), the lower limit of quantification was 2.00 ng·mL^-1 and 4.00 ng·mL^-1, respectively; the quality control samples with low, medium and high concentrations of Pro-PTX and PTX were within the batch, the relative standard deviation (RSD) between batches were all less than 9%; the relative deviation (RE) was within the range of ± 9%; In the stability test, both Pro-PTX and PTX in plasma were stable under various storage conditions. The method was sensitive, specific, and reproducible, and was suitable for the pharmacokinetic study of Pro-PTX in rats. Animal experiments were approved by the Ethics Committee of Laboratory Animal Management and Animal Welfare, Institute of Materia Medica, Chinese Academy of Medical Sciences (No.: 00003552).

OBJECTIVETo observe the efficacy of integrative medical sequential method in treating cerebral palsy (CP) children's intelligence development, muscular tension, serum interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-alpha).

METHODSTotally 111 CP children were randomly assigned to the control group (50 cases) and the treatment group (61 cases). All patients received comprehensive rehabilitation training and intravenous dripping of Monosialotetrahexosylganglioside Sodium Injection for 10 days. But those in the treatment group additionally received Chinese medical enema for brain resuscitation, relieving rigidity of muscles and activating collaterals for 14 days. Then they started another medication cycle and lasted for a total of 6 cycles. Serum IL-6 levels and TNF-alpha contents were determined before treatment. Scoring for muscular tension, Gesell score for intelligence development, contents of serum IL-6 and TNF-alpha were assessed before and after treatment in the two groups.

RESULTSCompared with before treatment in this group, muscular tension, Gesell scores for intelligence development all decreased in the two groups (P < 0.05). As for inter-group comparison, the decrement was more obvious in the treatment group than in the control group (P < 0.05). The total effective rate was 86.9% in the treatment group and 76.0% in the control group (P < 0.05). The contents of IL-6 and TNF-alpha were obviously reduced in the treatment group and the control group after treatment (P < 0.01). The decrement was more obvious in the treatment group (P < 0.05).

CONCLUSIONThe two treatment methods were effective for CP children, but the efficacy was superior in the treatment group than in the control group, indicating integrative medical methods could play a synergistic effect and optimize the treatment program for CP.

Cerebral Palsy ; drug therapy ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gangliosides ; therapeutic use ; Humans ; Infant ; Integrative Medicine ; Intelligence ; Interleukin-6 ; blood ; Male ; Phytotherapy ; Tumor Necrosis Factor-alpha ; blood

Objective To compare and analyze the similarities and differences of the blood perfusion characteristics of the renal pelvic carcinoma displayed by contrast-enhanced ultrasonography(CEUS) and contrast-enhanced computed tomography(CECT).Methods The characteristics of CEUS and CECT were anzlyzed retrospectively in 40 confirmed renal pelvic carcinoma cases by operation and pathology.The modality and phases of enhancement,including wash-in and washout time,as well as the perfusion appearances,were observed by CEUS and CECT.Results CEUS detected the cortical phase enhancement in all renal pelvic tumors,including synchronously enhancement in 14 tumors and delayed enhancement in 26 tumors.At peak times,hypoechogenicity compared to the normal renal cortex was shown in 31 tumors,hyperechogenicity in 4 tumors and isoechogenicity in 5 tumors.Fast wash-out in medulla phase was displayed in 35 tumors,isochronously wash-out in 2 tumors and delayed wash-out in 3 tumors.The minimum diameter in 40 renal pelvic tumors was 1.5 cm.CECT showed the enhancement in 38 tumors,mostly mild to moderate homogeneous enhancement,and enhancement was lower than the surrounding renal parenchyma.The diagnostic accuracy of CEUS was 77.5％ (31/40),and the diagnostic accuracy of CECT was 82.5 ％ (33/40).The difference was not statistically significant between CEUS and CECT ( P ＞0.05).Conclusions CEUS and CECT have higher diagnostic accuracy of the renal pelvic carcinoma,so the joint detection can increase the detection rate of early diagnosis of the renal pelvic carcinoma.