AIM: To develop a method to detemine pinostrobin in Weitengning Talets (Lindera reflexae Hemsl.) by HPLC. METHODS:A C_(18) column was used,methyl alcohol-water(80∶20) was used as a mobile phase and the wavelength of UV detector was set at 290 nm. RESULTS:The linearity of this method was good with the average recovery of 97.9%,RSD was 0.74%(n=5). CONCLUSION:The methed is simple,reliable,sensitivity,and with good reproducibility.It can be used in quality control of Weitengning Tablets.

OBJECTIVE: To explore the expression levels of three T-type calcium channel receptors (α1G; α1H; α1I) in the cochlea and spiral ganglion neurons of C57BL/6J mice with different ages. METHOD: Thirty cases of C57BL/6J mice were divided into three groups (6-8 W, 24-26 W, 42-44 W) according to the age. The expressions of three T-type calcium channel receptors were quantified by RT-PCR after hearing thresholds measured by ABR. RESULT: Three receptors were detected in the cochlea and spiral ganglion neurons of 6-8 W C57BL/6J mice. The quantitative results showed that the expression levels of α1H and α1I were highest among three receptors in spiral ganglion neurons and in the cochlea respectively. The expression levels of three receptors significantly decreased with age,especially at the age of 4244 W. CONCLUSION The expression of T-type calcium channel receptors reduced with age in the inner ear of C57BL/6J mice.
Animals ; Calcium Channels ; Calcium Channels, T-Type ; biosynthesis ; Cochlea ; metabolism ; Ear, Inner ; Mice ; Mice, Inbred C57BL ; Neurons ; Receptors, Calcium-Sensing ; Spiral Ganglion ; metabolism

This research is to establish an HPLC method for the simultaneous determination of ophiopogonin D, ophiopogonin D', ophiopogonin C, deacetylophiopojaponin A and ophiogenin-3-O-α-L-rhamnosyl-(1-->2)-β-D-glucoside in Ophiopogonis Radix. HPLC-ELSD analysis was performed on a Kromasil 100-5 C₁₈ column (4.6 mm x 250 mm, 5 µm), with the mobile phase of acetonitrile (A) -water (B) in gradient elution mode (0-45 min, 35%-55% A), at a flow rate of 1 mL · min⁻¹. The column temperature was 35 °C and the drift tube temperature was 100 °C in a gas flow rate of 3.0 L · min⁻¹. The result showed that baseline of all the 5 constituents was well separated, and every constituent had wide linearity range and good linear relation (r > 0.999). The recovery rate was between 95.75% and 103.1%. The new established method for simultaneous determination of saponin constituents in Ophiopogonis Radix was sensitive and has good, repeatability. It could be applied to quality evaluation of Ophiopogonis Radix.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Aim To explore the anti-apoptotic function of cardiac progenitor cells(CPCs)-derived exosome in vitro.Method CPCs were isolated from mouse heart using Magnetic Cell Sorting(MACS)system.Flow Cy-tometry(FC)determine the purity of stem cell surface antigen-1 positive(Sca-1 +)CPCs.Exosome was puri-fied from conditional medium,and confirmed by West-ern blot using CD63 as a marker,Nanoparticle Traffic-king Analysis(NTA)was used to detect the diameters and concentration of exosome.Then the cells were di-vided into control groups and CPC-exosome pre-protec-tion groups.H2 O2 was added into H9c2 cells to induce oxidative stress.Western blot was adopted to determine the expression of cleaved caspase-3.Results ① Im-munofluorescence showed that CPCs isolated by MACS were positively expressing Sca-1 protein;FC analysis showed that typical purity of Sca-1 +CPCs from the first
preparations was more than 95%.② WB demonstrated that CD63 of exosome isolated from CCMwas positively expressed,and NTA results showed that the diameters of exosome were (82.33 ±3.06)nm(n =3).Micro-scope detected PKH-26 labeled exosome appeared in the cytoplasma of H9c2 cells.③ Western blot showed the CPC-exosome pre-protection groups significantly down-regulated the levels of cleaved caspase-3 com-pared to the control groups(P <0.05).Conclusion CPC can secrete exosome which carries many important cargos,which can effectively gather in H9c2 cells. CPC-exosome can protect H9c2 cells from the oxidative stress induced by H2 O2 .Our results highlight a new perspective strategy for cardiac disease.

[ ABSTRACT] AIM:To investigate the role of encapsulated protein transfected into human bone marrow mesen-chymal stem cells ( hBMSCs) by polyethyleneimine ( PEI) , and to optimize the best mole ratio of PEI-proteins.METH-ODS:6 groups of DNase I-PEI complexes were constructed and the best mole ratio was explored by laser scattering analy-sis.The appearance of complexes was presented under transmission electron microscope.Meanwhile, 4 groups of construc-ted GFP-PEI complexes were utilized to transfect into the hBMSCs, which were isolated and expand in vitro.The fluores-cence intensity of transfected cells was observed under confocal microscope.In addition, the cytotoxicity of the complexes on the cell proliferation was detected by MTT assay.The activity of the intracellular proteins was testified by aβ-galactosi-dase staining experiment.RESULTS:When the mole ratio of PEI and protein was adjusted to 4∶1, the complex transfec-tion efficiency was the best, and β-galactosidase color test turned blue.CONCLUSION:PEI has the character of encap-sulating various proteins to nano-complexes.The proteins transfected into bone marrow mesenchymal stem cells are con-firmed to have functional activity.As a protein carrier, PEI is of high efficiency and low toxicity, thus providing a new way for stem cell reprogramming.

Objective To quantitatively assess the cardiopulmonary exercise function of spinal cord injury (SCI) patients and observe the effect of aerobic exercise on cardiopulmonary function, motor function and activities of daily living. Methods From December, 2014 to June, 2016, 34 incomplete SCI patients (ASIA C and D) and 23 healthy controls received cardiopulmonary exercise test (CPET). SCI pa-tients were randomly divided into conventional rehabilitation group (n=17) and aerobic exercise group (n=17). The aerobic exercise group received aerobic exercise for four weeks. They were assessed with CPET, motor and sensory function, walking index for spinal cord injury II (WISCI II) and spinal cord independence measure (SCIM) before and four weeks after training. Results Oxygen uptake (VO2)peak, anaerobic threshold (AT), metabolic equivalent of energy (METpeak), VO2/heart rate (HR)peak, respiratory exchange rate (RER)peak, minute ventilation (VE)peak, work rate (WR)peak and systolic blood pressure (SBP)peak were lower in the patients than in the controls (t>2.714, P<0.05). VO2peak、AT、METpeak、VO2/HRpeak、WRpeak increased in the aerobic exercise group after training (t>2.431, P<0.05). METpeak and WRpeak improved in the conventional rehabilitation group after training (t>3.282, P<0.01). The scores of motor in ASIA and SCIM improved in both groups after training (t>2.985, P<0.05). Conclusion The cardiopulmonary function decreased in incomplete SCI patients, which could be improved by moderate intensity aerobic exercise.

Near-infrared (NIR) diffuse reflectance spectroscopy was used to analyze the impact of multi-class particle size of Chinese material medica (CMM) based on the spectral characteristics in overtone and combination region. Several types of CMM (60, 80, 100,120 mesh) were subjected to NIR spectra analysis. Spectral reproducibility was examined after sample repackage. The result showed that the effects of particle size on the NIR spectra were different according to different bands, in the combination region and first combination-overtone region. Spectroscopy intensity was proportional to the particle size and influence of particle size was greater as the wavelength increased. While in the second combination-overtone region, it was inversely proportional to particle size. To the sampling loading error, the result indicated that when the mesh number was larger than 60 mesh, the error was small. The appropriate particle size was clarified to guarantee the accuracy and reliability of NIR diffuse reflectance spectroscopy in CMM.
Drugs, Chinese Herbal ; analysis ; Materia Medica ; analysis ; Particle Size ; Spectroscopy, Near-Infrared ; methods

The on-line monitor for the changes in the content of baicalin in Scutellariae Radix formula particles during the extraction process was conducted by using near infrared spectroscopy (NIR). High performance liquid chromatography (HPLC) was used as a reference method. Kennard-Stone (KS) was used to divide sample sets, so as to compare different pretreatment methods. The synergy interval partial least squares (SiPLS) was used to screen out modeling wave band to establish partial least-squares models. The relative error method was applied to predict forecast set samples of Scutellariae Radix in three extraction phases. The results showed that the model established by Savitzky-Golay smoothing with 11 points (SG11 points) was the best, with the root mean square with cross validation (RMSECV), root mean square error of correction (RMSEC) and root mean square error of prediction (RMSEP) of baicalin of 0.092 7, 0.134 4 and 0.114 8, respectively, the determination coefficient R2 of greater than 0.99, and the relative error of baicalin content of less than 5%. This indicates that the on-line near infrared reflectance spectroscopy could be applied in on-line monitor and quality control of the extraction process of Scutellariae Radix formula particles.
Automation ; methods ; Drugs, Chinese Herbal ; chemistry ; standards ; Quality Control ; Scutellaria baicalensis ; chemistry ; Spectroscopy, Near-Infrared ; methods

Near-infrared (NIR) was used as rapid analysis method to identify the sulfur fumigated Puerariae Lobatae Radix. NIR spectra of the cross-sectional and longitudinal selection of samples were acquired. Principal component analysis was conducted. The samples were randomly selected. The different pretreatment methods were compared. Discriminant models were established for every type of spectra to calculate the recognition rate. The orthogonal test and nonparametric test were used to test data normality. The result showed that absorbance values of different sections were different due to the different structure, and the raw spectra were analyzed by PCA method. The result founded that the cumulative contribution rate was arrived at 99.2% while the PC numbers were arrived at 3. The pretreatment method based on the MSC + 1D + Savitzky-Golay was the best to establish the model. For the 50 models constructed with cross-section and longitudinal spectra and total spectra, the recognition rate were (94.4 ± 0.66)%, (94.4 ± 0.66)%, (95.3 ± 0.65)%, respectively, and no difference was observed. The NIR method could be used to identify the sulfur fumigated Puerariae Lobatae Radix.
Chemistry, Pharmaceutical ; methods ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Fumigation ; Pueraria ; chemistry ; Spectroscopy, Near-Infrared ; Sulfur ; chemistry