The pharmacokinetics of a long-circulating PEGylated Radix Ophiopogonis polysaccharide (ROP) was investigated in rats following i.v. or s.c. administration at three dose levels (9, 20, 50 mg x kg(-1)). A moderate coupling reaction between the hydroxyl-activated ROP and the amino-terminated mPEG was chosen to produce PEGylate ROP. The grafting degree of the prepared conjugate was 1.03, and the molecular mass of mPEG used was 20 kDa. High-performance gel permeation chromatorgraphy with fluorescein isothiocyanate prelabeling was established to determine levels of the conjugate in plasma. The results showed that the elimination half-life of the conjugate following s.c. administration was basically identical to that after iv administration. An accurate linear correlation was observed between administration doses and areas under the curve of plasma conjugate level vs. time profile, regardless of the administration route. The absolute bioavailability of the conjugate following sc administration was approximately 56%, and the mean in vivo residence time was 52.1 h, increased 2.4 times compared to those of iv administration. In general, linear pharmacokinetics was observed for the conjugate within the dose range studied, and sc should be a promising administration route for the conjugate.
Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Half-Life ; Injections, Intravenous ; Injections, Subcutaneous ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polyethylene Glycols ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

To evaluate flow-cytometry and Nageotte method for counting residual WBC in thrombocytaphoresis concentrates, their accuracies were determined by dilution studies separately; the repeatability was determined by measuring the interassay coefficient of variation for 14 replicates of a sample with known leukocyte concentration. 102 samples of leukocyte-depleted thrombocytaphoresis concentrates were detected by the above mentioned two methods, and the results were compared each other. The results showed that no difference was observed between two methods over a range of leukocyte concentrations from 0.8 to 10 WBC/microl (P > 0.05). In conclusion, flow-cytometry and Nageotte methods can be used for quality control of WBC-reduced blood components.
Blood Component Transfusion ; Evaluation Studies as Topic ; Flow Cytometry ; Humans ; Leukocyte Count ; methods ; Leukocyte Reduction Procedures ; methods ; Plateletpheresis

OBJECTIVEThe choice of surgical approaches for salvage surgery based on the location and invasion of recurrent and residual lesions of nasopharyngeal carcinoma (NPC), surgical results, complications, and survival were assessed.

METHODSThirty-seven cases with recurrent and residual lesions of NPC underwent salvage surgery between March 1991 and January 2005 were analysed retrospectively. Of 37 patients, 23 were men and 14 women, with a median age of 46.5 years (26 - 57 years); 4 were at stage I, 10 at stage II, 14 at stage III, and 9 at stage IV; 5 cases were with cervical metastasis, including 3 cases of N1 and 2 cases N2. All recurrent and residual lesions of NPC were determined by biopsy. On the location and invasion of recurrent and residual lesions of NPC, 8 cases underwent endoscopic resection of lesions, 12 cases of the palate nasopharyngectomy, 5 cases of maxillary swing, 4 cases of maxillary swing plus prerenal approach, 2 cases of lateral rhinotomy plus coronal flap approach, and 6 cases transfacial plus nasal pyramid swing approach. Five cases with cervical metastasis received neck dissection in addition to the operations for recurrent and residual lesions of NPC. Postoperatively 31 cases received radiotherapy with dosage of 60 Gy, among them 15 cases with concurrent chemoradiation therapy, and 6 cases with clear surgical margin did not received radiotherapy or chemotherapy. The cases were followed up for 12 - 72 months, with a median of 45 months.

RESULTSTotal resection for the recurrent and residual lesions of NPC accounted for 91.8% (34/37) and subtotal resection for 8.2% (3/37). The accident of perioperative complications was 24.3% (9/37). The 3- and 5-year overall disease-free survival rates (DFSR) were 62.1% and 43.3%, respectively. The 3- and 5-year overall survival rates (OSR) were 72.9% and 51.3%, respectively. The 5 year DFSR of cases at stage I-IV were 100%, 40%, 28% and 11% (χ(2) = 10.0, P < 0.01), respectively. The 5 year OSR were 100%, 70%, 35% and 28% (χ(2) = 11.5, P < 0.01), respectively.

CONCLUSIONSSalvage surgery is a justified treatment for the recurrent and residual lesions of NPC, by which some patients with recurrent and residual lesions of NPC can be salvaged.

Adult ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Neoplasm Recurrence, Local ; surgery ; Neoplasm Staging ; Neoplasm, Residual ; Prognosis ; Retrospective Studies ; Salvage Therapy ; methods

OBJECTIVETo observe the effect of ligand of peroxisome proliferators-activated receptor gamma (PPAR gamma) 15d-PGJ2 on the proliferation and activation of hepatic stellate cells (HSC) and to study the role played by PPAR gamma during the process of HSC activation.

METHODSBy using RT-PCR and cell culture, we investigated the effects of 5 micro mol/L and 10 micro mol/L 15d-PGJ2 on culture-activated HSC and on PDGF-induced HSC proliferation, production of extracellular matrix and expression of chemokines.

RESULTSThe expression of alpha-SMA was significantly suppressed by 5mumol/L 15d-PGJ2, and the expression of PPAR gamma was significantly higher in the 15d-PGJ2 treated group than in the untreated group (0.64+/-0.03 vs 0.09+/-0.01, t=36.0517, P<0.01); PDGF-induced HSC proliferation was dose-dependently suppressed by 15d-PGJ2; the expressions of PPAR gamma in 5 micro mol/L and also in 10 micro mol/L 15d-PGJ2 plus PDGF pre-treated group increased much more than those in the PDGF-treated group (0.03+/-0.02 vs 0.60+/-0.03, t=42.6616, P<0.01 and 0.03+/-0.02 vs 0.69+/-0.04, t=33.83, P<0.01); the expressions of alpha-SMA, alpha 1 (I)-collagen and MCP-1 were suppressed.

CONCLUSIONActivation of PPAR gamma can modulate pro-fibrotic and pro-inflammatory roles of HSC and the increased expression of PPAR gamma may become a new target for antifibrosis.

Animals ; Cell Differentiation ; Cell Proliferation ; drug effects ; Cells, Cultured ; Hepatic Stellate Cells ; cytology ; metabolism ; Male ; PPAR gamma ; metabolism ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVEIsolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF.

METHODSColo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical. To induce cell differentiation, tumour spheres were cultivated without EGF and bFGF in the presence of 10% serum. Then we analysed expressions of stem cell surface markers CD133 and CD44 among undifferentiated cell, post-differentiated cells and routine Colo205 cells under serum-supplemented culture condition by flow cytometry. At last we compared cell cycle and spectral karyotype between two groups.

RESULTSIn SFM consisting of EGF and bFGF, a minority of Colo205 cells could survive, proliferate and form the suspended tumor spheres. We detected high Musashi-1 expression in these cells. Compared with the SSM group and the post-differentiation SFM group, the expressions of CD133 and CD44 were significantly increased in the undifferentiated SFM group (P<0.05). There was no statistical difference in the expression of CD133 and CD44 between the post-differentiation SFM group and the SSM group (P>0.05). Cell cycle analysis indicated that tumor spheres were of a high proliferation state.We could not find any noticeable difference in the number of chromatosomes between the SFM group and the SSM group.

CONCLUSIONTumor spheres in which enriched cancer stem cells can be generated under serum-free culture condition with EGF and bFGF.

AC133 Antigen ; Antigens, CD ; metabolism ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Cell Proliferation ; Culture Media, Serum-Free ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Neoplastic Stem Cells ; cytology ; metabolism ; Nerve Tissue Proteins ; metabolism ; Peptides ; metabolism ; RNA-Binding Proteins ; metabolism ; Spheroids, Cellular ; cytology

Confocal laser scanning microscopy(CLSM) is a new technique for microscopic imaging, which can collect the transverse section image of the samples and produce three-dimensional reconstruction and present higher spatial resolution than the conventional light microscope. As a precision instrument for the microscopic image, it plays an important role in forensic pathology. The article reviews the recent research achievements from sudden cardiac death, bullet wound and nervous system damage, etc, and explores the potential applications of the forensic pathology research and forensic practice.
Calcium/metabolism* ; Craniocerebral Trauma/pathology* ; Death, Sudden, Cardiac/pathology* ; Forensic Pathology/methods* ; Humans ; Image Processing, Computer-Assisted ; Microscopy, Confocal ; Myocardium/ultrastructure* ; Sensitivity and Specificity ; Trauma, Nervous System/pathology* ; Wounds and Injuries/pathology* ; Wounds, Gunshot/pathology*

Two new phenylpropanoids(1 and 2), together with thirteen known compounds(3-15), have been isolated from the root of Paeonia lactiflora by using various chromatographic techniques. Their structures were identified by spectroscopic data analysis(MS,IR,1D and 2D NMR)as(+)-(7R,8R)-1-guaiacyl-1,2-propanediolacetonide(1),(-)-(7R,8S)-1-guaiacyl-1,2-propanediolacetonide(2),O-senecioyllomatin(3),O-angeloyllomatin(4),(+)-cis-3'-senecioyloxy-4'-angeloyloxy-3',4'-dihydroseselin(5),columbianadin(6), benzyl 2,5-dihydroxybenzoate(7),3,6-dimethyl-5-hydroxyBenzo-furan(8),(S)-evofolin-A(9),2,3-dihydroxy-4-methoxyacetophenone(10), 2,5-dihydroxy-4-methoxyacetophenone(11), 2,5-dihydroxy-4-methyl acetophenone(12),ethyl 4-hydroxybenzoate(13), vanillic acid(14),and 4-hydroxy-3-methoxybenzaldehyde(15).Compounds 1 and 2 were new compounds,and compounds 3-9 were obtained from the genus Paeonia for the first time.

Twenty five known aromatic glycosides (1-25) and three known sesquiterpene glycosides (26-28) have been isolated from the twigs of Litsea cubeba by using various chromatographic techniques. Their structures were identified by spectroscopic data analysis (MS, IR, 1D and 2D NMR) as (7S,8R)-dehydrodiconiferyl alcohol 4,9'-di-O-β-D-glucopyranoside(1),(7S,8R)-5-methoxydihydrodehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside(2), (7S,8R)-urolignoside(3), (7R,8S)-dihydrodehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside(4), saposide B(5), lanicepside A(6), matairesinol-4-O-β-D-glucopyranoside (7), tyraxjaponoside B(8), (+)-lyoniresinol-9'-O-β-D-glucopyranoside (9), alaschanisoside A (10), syringin (11), psoralenoside (12), isopsoralenoside (13), scopolin(14), 2,6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside (15), 3-hydroxy-4,5-dimethoxyphenyl-β-D-glucopyranoside (16), 2-(3,4-dihydroxyphenyl)ethyl-β-D-glucopyrnoside (17), 2-(4-dihydroxyphenyl)ethyl-β-D-glucopyranoside (18), (+)-catechin-7-O-β-D-glucopyranoside (19), 3'-O-methylepicatechin-7-O-β-D-glucopyranoside (20), kaempferitrin (21), quercetin-3-O-α-L-rhamnopyranside (22), kaempferol-3-O-β-D-glucopyranoside (23), kaempferol 3-O-β-D-glucopyranosyl(1→2)-O-β-D-galactopyr anoside-7-O-α-L-rhamnopyranoside (24), quercetin 3-O-α-L-rhamnopyranosyl(1→6)-O-β-D-glucopyranosyl(1→3)-O-α-L-rhamnopyranosyl(1→2)-O-β-D-glucopyranoside (25), staphylionoside D(26), vomifoliol 9-O-β-D-glucopyranoside (27), dihydrovomifoliol-O-β-D-glucopyranoside (28). Compounds 1-21 and 24-28 were obtained from this genus for the first time.