Objective: To prepare sinomenine hydrochloride (SIN-HCl) liposomes with high entrapment efficiency (EE) and to illustrate the effects of drug quantity and particle size on EE. Methods: Centrifugation sedimentation-centrifugation ultrafiltration was employed to determine EE of liposomes. Thin film hydration (TFH), reverse phase evaporation (REV), and ether injection (EI) were screened based on EE and formability of liposomes. The effects of water type, pH value, ion concentration of hydration liquid, pH gradient active drug loading, lecithin-cholesterol ratio, and drug-lipid ratio on EE of liposomes were investigated. The relationship between EE and the factors affecting the drug quantity and particle size was probed with a comprehensive design experiment. The stability of typical liposomes was evaluated at 4 °C. Results: The optimal preparation technology was TFH for SIN-HCl liposomes and citrate buffer solution (CBS) was the best hydration liquid. The liposome EE increased with the increase of pH values of CBS. When the pH value of CBS was fixed, the EE increased as a result of decrease in the ion concentration of CBS. pH gradient active drug loading led to increase of EE. The preferable hydration liquid for liposomes was CBS with pH value of 2.5. The optimal ratio of soybean lecithin to cholesterol was 6:1. Increasing ratios of SIN-HCl to soybean lecithin from 1:6 to 6:6 led to a slight decrease in EE of liposomes without probe signification. A quantitative relationship was established between the EE and drug quantity and liposome size. The EE of SIN-HCl liposomes prepared by certain particle size and drug quantity could reach over 80%. The typical liposomes showed a good stability. Conclusion: The technology of pH gradient active drug loading is able to prepare SIN-HCl liposomes with high EE.

Objective: To develop a method for determining the entrapment efficiency of sinomenine hydrochloride (SM-HCl) liposomes and to illuminate the drug retention property in the liposomes. Methods: Thin film hydration method was employed to prepare SM-HCl liposomes. HPLC was used to determine drug content of the liposomes. A Kromasil ODS C18 column (250 mm x 4.6 mm, 5 μ) was used with an isocratic elution composed of methanol, water, and ethylenediamine in the ratio of 55 : 45 : 0.225 at a flow rate of 1.0 mL/min. The column was maintained at 30 °C. The UV detector was set at 265 nm. Centrifugation sedimentation combined with centrifugation ultrafiltration was used to determine drug entrapment efficiency of the liposomes. The entrapment efficiencies of an SM-HCl liposome sample (hydrated with citric buffer solution at pH 7.0) and its diluted sample were compared. Results: The pharmaceutical excipients and solvents for analysis had no interference with the determination of sinomenine. Sinomenine had a good linear relation in the range of 9.82-78.6 μg/mL (r = 0.999 7), the intra-day and inter-day precisions were with RSD≤2.1% and the averaged recovery was within 99.29%-100.8%. SM-HCl solution (50 μL) was able to saturate the drug absorption of ultrafiltration films. The entrapment efficiencies of the SM-HCl liposome sample (hydrated with citric buffer solution at pH 7.0) and its double-volume diluted sample were 33.16% and 14.75%, respectively. Conclusion: HPLC and centrifugation sedimentation combined with centrifugation ultrafiltration are able to determine the entrapment efficiency of SM-HCl liposomes efficiently and accurately. Initial filtrate (50 μL) should be discarded in the process of ultrafiltration in order that the drug concentration in filtrate may be equal to that of external aqueous phase of liposomes. The retention of sinominine in the liposomes is poor, although it has considerable affinity to the lipid bilayers.

A method was proposed for the separation and determination of paraffin waxes in food by HPLC-evaporative light scattering detection (ELSD). A normal-phase column was used to separate nonparaffinic and paraffinic materials without resolving the latter into individual components. The t-test method was adopted for the evaluation of mean difference between response factors of n-alkanes in paraffin waxes on ELSD detector. No mean difference was obtained between response factors, which can be used for quantitative determination of paraffin waxes in food. The determination results obtained by HPLC-ELSD were compared with those by GC-MS. The linear range for the determination of paraffin waxes was in the range from 10 to 500 mg/L with a correlation coefficient of 0.9988, and the limit of detection was 1.0 mg/L. With the spiking level of 10, 50 and 100 mg/kg, the recovery ranged from 84.6% to 105.4% and the relative standard deviation ranging from 5.4% to 7.2%. The proposed method is simple, fast and sensitive.

OBJECTIVETo develop an HPLC method to determine vitexin-rhamnoside in plasma of Beagle dogs and study the pharmacokinetics and bioavailability of Yixintong sustained release tablets in Beagle dogs.

METHODA newly-developed HPLC method using C18 column and methanol-acetonitrile-tetrahydrogenfuran-0.5% acetic acid (1:1:19.4:78.6) as mobile phase was validated, and then was employed to determine vitexin-rhamnoside in plasma of Beagle dogs after oral administration of Yixintong sustained release tablets and general tablets. The main pharmacokinetic parameters were estimated by pharmacokinetic program 3p87. The non-compartmental pharmacokinetic parameters were also calculated on basis of the statistic moment theory.

RESULTThe pharmacokinetic profiles of Yixintong sustained release tablets and the general tablets were fitted to a one-and two-compartment open model, respectively. The T1/2, Tmax, AUC0-infinity and MRT for Yixintong sustained release tablets were 5.22 h, 4.0 h, 6,792.75 ng x h x mL(-1) and 8.4 h, respectively, compared with 8.94 h, 1.0 h, 5,880.4 ng x h x mL(-1) and 6.1 h for the general tablets. The relative bioavailability of the Yixintong sustained release tablets was 115.5% in Beagle dogs.

CONCLUSIONThe sustained-release characteristic of Yixintong sustained release tablets were confirmed by pharmacokinetic study.

Administration, Oral ; Animals ; Apigenin ; chemistry ; Biological Availability ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Female ; Male ; Plasma ; chemistry ; drug effects ; Tablets ; administration & dosage ; pharmacokinetics

AIMTo prepare ondansetron hydrochloride osmotic pump tablets (OND-OPT) and investigate their in vitro drug release behavior.

METHODSOND-OPT were prepared with a single punch press and pan coating technique. Osmotic active agents and plasticizer of coating film were chosen by drug release tests. The effects of the number, position and direction of drug release orifice on release behavior were investigated. The relation between drug release duration and thickness of coating film, PEG content of coating film and size of drug release orifice was established by uniform design experiment. The surface morphological change of coating film before and after drug release test was observed by scanning electron microscopy. The osmotic pumping release mechanism of OND-OPT was confirmed by drug release test with high osmotic pressure medium.

RESULTSLactose-mannitol (1:2) was chosen as osmotic active agents and PEG400 as plasticizer of coating film. The direction of drug release orifice had great effect on the drug release of OND-OPT without HPMC, and had no effect on the drug release of OND-OPT with HPMC. The OND-OPT with one drug release orifice at the centre of the coating film on one surface of tablet released their drug with little fluctuation. The drug release duration of OND-OPT correlated with thickness of coating film and PEG content of coating film, and didn't correlate significantly with the size of drug release orifice. OND-OPT released their drug with osmotic pumping mechanism predominantly.

CONCLUSIONOND-OPT are able to realize ideal controlled drug release.

Antiemetics ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Hypromellose Derivatives ; Lactose ; Mannitol ; Methylcellulose ; analogs & derivatives ; Ondansetron ; chemistry ; Osmotic Pressure ; Polyethylene Glycols ; Solubility ; Tablets ; beta-Cyclodextrins

This paper reviews the lastest progress on oral prolonged-release preparation of traditional Chinese medicine. Four materials, include component, effective parts, single drug, and compound drugs of traditiong Chinese medicine, have been used to produce oral prolonged-release preparation. The main contents are study of preparation and evaluation of in vitro release. There are also some research works on integrative evaluation, pharmacokinetics and pharmacological activity of the prolonged-release preparation. It believes that the study on oral prolonged-release preparation of traditional Chinese medicine will have good prospect.
Animals ; Delayed-Action Preparations ; pharmacokinetics ; toxicity ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; toxicity ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo investigate the effect and influence factors on knee joint peripheral fractures and/or dislocations with an associated vascular injury through retrospectively study.

METHODSFrom March 2002 to November 2007 31 patients with knee joint peripheral fractures and/or dislocations with an associated vascular injury were treated, including 24 males and 7 females with a mean age of 41 years (range from 21 to 62 years). Definite diagnosis of vascular injury by combining colored ultrasonic, CTA, operative exploration with clinical signs, fixing fractures and/or dislocations with fixators, plates and screws, reconstructing blood circulation based on the condition of the vascular injury by vascular repair, homograft vein or artificial vascular grafting separately and analysing the effects of PSI, diagnosis and treatment methods on salvage lower extremities.

RESULTSSuccessful reconstruction was carried out in 31 cases, however there were 1 death because of mult-fractures and brain injury and 6 amputation, 24 cases successful salvage followed up mean 24.2 months, 6 cases bone nonunion and infected bone defect were cured by delayed bone planting or bone transportation. Ligaments repair reconstruction of 7 cases knee joint dislocation were done in delayed 3 or 4 weeks after first operation, the good functional rate was 71.4%.

CONCLUSIONSThe patients of PSI under 10 grades in knee joint peripheral fractures and/or dislocations with an associated vascular injury should been carried out treatment, early definite diagnosis and blood circulation reconstruction are the key factors of successful salvage treatment.

Adult ; Female ; Follow-Up Studies ; Fractures, Bone ; complications ; diagnosis ; surgery ; Humans ; Knee Dislocation ; complications ; diagnosis ; surgery ; Knee Injuries ; diagnosis ; surgery ; Male ; Middle Aged ; Popliteal Artery ; injuries ; Retrospective Studies ; Treatment Outcome ; Vascular Grafting ; Vascular System Injuries ; complications ; diagnosis ; surgery ; Young Adult

The aim of study was to set up a suitable method of isolation, culture and identification of endothelial progenitor cells (EPC) derived from rabbit bone marrow. Density gradient centrifugation was used to isolate mononuclear cells from bone marrow, the isolated mononuclear cells were cultured with specific culture medium for EPCs. EPCs were identified by cellular morphologic observation, immunohistochemistry testing, flow cytometry and the function test of taking up Dil-ac-LDL and FITC-UEA-1. The results indicated that the newly isolated bone marrow-derived mononuclear cells exhibited a round appearance, following culture for 48 hours, adherent cells grew in colony cluster, presenting with round or irregular appearance, and nuclear division was obvious. On day 7, flaky cell colonies mutually connected together, presenting with spindle-shaped cells. Immunohistochemistry testing in the EPCs showed CD133(+), CD34(+), VIII factor(++), KDR(++); flow cytometry testing showed that the positive rate of CD133 was (18.23+/-7.12)%, the positive rate of CD34 was 47.71+/-14.85%, the positive rate of CD31 was (71.61+/-13.51)%, the positive rate of KDR was (87.24+/-11.40)%. And more than 80% EPC could take up both Dil-acLDL and FITC-UEA-1. It is concluded that the mononuclear cells isolated from bone marrow by density gradient centrifugation can differentiate into EPCs under special culture situation.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; cytology ; Rabbits ; Stem Cells ; cytology

OBJECTIVETo identify protein markers for the early diagnosis of pancreatic cancer by a comparative proteomic method.

METHODSComparative analysis on the pancreatic peripheral blood protein profiling from 20 pancreatic cancer patients, 10 chronic pancreatitis patients and 20 cancer-free controls from May 2007 to September 2008 was carried out by two-dimensional fluorescence electrophoresis (2D-DIGE). Differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The significance difference proteins were confirmed by Western-blot.

RESULTSA differentially expressed proteins: complement 3 (C3) was identified. The gray level of C3 in pancreatic cancer tissue, chronic pancreatitis, and normal control group were 1.63 ± 0.28, 0.65 ± 0.13 (t = 11.81, P = 0.00) and 0.88 ± 0.19 (t = 9.93, P = 0.00), respectively. C3 was high expression in pancreatic cancer group compared with normal control group. The expression of C3 was higher in pancreatic cancer group than in chronic pancreatitis group. The high expression of C3 in pancreatic carcinoma was confirmed by Western blot.

CONCLUSIONS2D-DIGE and MALDI-TOF-MS technology is a quick, easy and practical method to screen for specific biomarkers in serum of patients with pancreatic carcinoma. The identified protein C3 in this study may be as specific serum biomarkers of pancreatic carcinoma.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Case-Control Studies ; Complement C3 ; analysis ; Early Diagnosis ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; Pancreatitis, Chronic ; blood ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Two-Dimensional Difference Gel Electrophoresis

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