Adipocytes ; cytology ; Animals ; Cell Proliferation ; Cells, Cultured ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology

OBJECTIVETo establish the method for determining polysorbate 80 in Reduning injection by HPLC-ELSD, and to control the mass of polysorbate 80 in Reduning injection.

METHODIt was performed by HGPC-ELSD with TOSHTSK-GEL G4000PWxl (7.8 mm x 300 mm, 10 μm). Water was used as mobile phase, the flow rate was 0.7 mL x min(-1), and the temperature was set at 30°C. The evaporated light scattering detector was adopted. The drift tube temperature was 55°C, and nitrogen was used as carrier gas, with the flow rate of 2.0 L x min(-1) and gain of 1.0.

RESULTThe calibration curve showed good linearity of polysorbate 80 in the test range from 1.01 to 15.20 g x L(-1) (r2 = 0.999 3). The recovery rate was 98.10% with RSD of 2.0%.

CONCLUSIONThe method is simple, rapid, accurate and reliable and suitable for the determination of polysorbate 80 in Reduning injection.

Calibration ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Injections ; Polysorbates ; analysis ; Reproducibility of Results ; Time Factors

This study was aimed to acquire the dynamic changes of chlorogenic acid and luteoloside of honeysuckle at different collecting periods in order to decide the best harvesting time of honeysuckle in Donghai County, Jiangsu Province. The content determination method used in the detection of chlorogenic acid and luteoloside of honeysuckle was from the 2010 edition of the Chinese Pharmacopoeia. The skills of HPLC fingerprint characteristic features, and the yield of pressed flowers were combined in the evaluation of honeysuckle at the three-green period, two-white pe-riod, great-white period, silver-flower period and the golden-flower period. The results showed that the content of honeysuckle at different blossoming stages had obvious changes in content of chlorogenic acid and luteoloside, as well as the pressed flower quality and yield of the flower. It was concluded that the optimal harvest time of honey-suckle was for the two-white period and the great-white period, which was consistent with the real origin.

Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

Aim To investigate the effects of U50,488H(a selective ?-opioid receptor agonist)on ventricular arrhythmias induced by myocardial ischemia and reperfusion in rats and to elucidate their mechanisms.Methods The contents of CK(creatine phosphokinase)and LDH(lactate dehydrogenase)were measured; The isolated heart was perfused using langendorff equipment. Heart rate(HR), arterial blood pressure(ABP), left ventricular pressure (LVP), cardiac function (?dp/dtmax), rate of ventricular tachycardia(VT)and ventricular fibrillation(VF)were examined in rats in vitro. The incidence of ventricular arrhythmias and arrhythmia score were also determined; The change of sodium currents (INa) induced by U50,488H was detected by whole-cell recording mode using patch clamp.Results ① In comparison with I/R group, the contents of CK、LDH in plasma of rats in U50,488H+I/R group were significantly lowered(P

OBJECTIVETo establish an analytical method for the fingerprint of triterpenoid constituents of Poria by HPLC and compare the fingerprints of different medicinal parts of Poria in order to provide basis for controlling Poria quality.

METHODThe HPLC chromatographic conditions were Waters Symmetry C18 column (4.6 mm x 250 mm, 5 μm), 0.1% phosphoric acid (A) and acetonitrile (B) as gradient mobile phases, flow rate being 1.0 mL x min(-1), column temperature at 30 degrees C, The detection wavelength was set at 210 nm; The cluster analysis was carried on by SPSS 15.0.

RESULTThe HPLC fingerprints of triterpenoid constituents of Poria were set up. There were 16 common peaks in different medicinal parts. The results of method validation met technical requirement of fingerprints; Triterpenoid constituents in White Poria and Poria cum Radix Pini were different from Poria. The content of pachymic acid was the highest in Poria. The effect of habitat on the quality was no obvious difference.

CONCLUSIONThe method is stable, reliable, reproducible, and can be used as an effective means of Poria quality evaluation.

Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Poria ; chemistry ; Triterpenes ; analysis

Taking guizhi fuling capsule (GZFL) for instance, a new method about reference Chinese medicine preparation which was used as standard substance for the quality evaluation of complex Chinese medicine preparation by the fingerprint of reference preparation instead of standard fingerprint was proposed. It could eliminate the errors from different instruments, chromatographic columns and solve the problem of similarity matching in the absence of standard fingerprint. The qualification of reference GZFL was evaluated according to the quality control method of GZFL from Chinese Pharmacopoeia. Then multiple batches of GZFL were estimated, taking fingerprint of reference preparation and standard fingerprint as references, respectively, at different instruments and chromatographic columns. Finally, the packaging and expiration date for reference GZFL were confirmed according to the results of stability investigation. The results indicated that the fingerprint of reference GZFL could be used to assess the quality of GZFL better than standard fingerprint. The data of accelerated stability and long-term stability test demonstrated that reference GZFL was stable in the conditions of double blister package. Therefore, reference GZFL can be used as standard substance in quality control of GZFL.
Capsules ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; standards ; Reference Standards

Near infrared (NIR) spectroscopy as a kind of rapid process analysis technology has been successfully applied in Chinese medicine pharmaceutical process. In this research, the technology was adopted to establish the rapid quantitative analysis models of main indicators from the Lonicera japonica and Artemisia annua alcohol precipitation process of Reduning injection. On-line NIR spectra of 142 samples from alcohol precipitation process were collected and the content of main indicators for each sample were detected through off-line HPLC. With eliminating outliers, determination of spectra pretreatment method and selecting optimal band, the NIR quantitative calibration model for each indicator was established using partial least squares (PLS). These models were used to predict the unknown samples from precipitation process of Reduning injection to achieve the goal of rapid detection. The results showed that the models were ideal. The correlation coefficients of models for neochlorogenic acid, chlorogenic acid, 4-O-caffeoylquinic acid and secoxyloganin were 0.973 872, 0.985 449, 0.975 509 and 0.979 790, respectively and their relative standard errors of prediction (RSEP) were 2.922 49%, 2.341 37%, 2.930 40% and 2.184 60%, respectively. This study indicated that the NIR quantitative calibration model showed good stability and precision, and it can be used in rapid quantitative detection of main indicators of efficacy in order to on-line monitor the alcohol precipitation process of Reduning injection.
Chemical Precipitation ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Lonicera ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods

AIMTo study the pharmacokinetics of spinosin in rat plasma after oral administration of Suanzaoren extract using sulfamethoxazole (SMZ) as internal standard by RP-HPLC method.

METHODSPlasma samples were deproteined with acetonitrile, followed by evaporation of the acetonitrile to dryness. The residual was then resolved in mobile phase and HPLC separation was achieved on a Hypersil C18 (5 microns, 200 mm x 4.6 mm ID) column at 35 degrees C. The mobile phase consisted of acetonitrile-water-acetic acid (15:85:1) at a flow rate of 0.7 mL.min-1. The UV detection wavelength was set at 334 nm.

RESULTSThe calibration curve was shown to be linear over the range from 18.1 to 903.5 micrograms.L-1 (r2 > or = 0.995). Mean recovery was 94.5%. Within-day and between-day precisions RSD were less than 9.0%. The limit of quantitation was 18.1 micrograms.L-1. The plasma spinosin was stable at -20 degrees C.

CONCLUSIONThe simple, sensitive and accurate HPLC method developed has been applied to determine the pharmacokinetics of spinosin in rat plasma after having taken Suanzaoren extract at a single dose.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Flavonoids ; blood ; isolation & purification ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Seeds ; chemistry ; Ziziphus ; chemistry

OBJECTIVETo study the effectiveness of sequential occlusal adjustment for kinetic food impaction.

METHODS36 patients who claiming food impaction with normal proximal contact were examined and analyzed about their occlusal relationship and configuration. Sequential occlusal adjustment was made to reduce pestle-mortar-like cusp, to create food escaping groove and to decrease mesial surface of the distal tooth cusp. One week, two weeks and six months later, the patients were reexamined and their oral conditions were evaluated.

RESULTSAn elimination of food impaction was claimed for 32 patients in one week appointment and all 36 patients in two-week appointment. Six months later, no patient reported food impaction.

CONCLUSIONThe use of sequential occlusal adjustment results in an effective elimination of some kind of kinetic food impaction.

Food ; Humans ; Mastication ; Occlusal Adjustment ; Tooth