AIMTo study the effect of hyaluronic acid chitosan-based microemulsion (HAC-ME) on the permeability of blood brain barrier( BBB) by using Evans blue (EB) as the indicator.

METHODSA formamide extraction-ultraviolet spectrophotometry method was employed to determine the concentrations of EB in each of the tissues. The in vivo distribution of HAC-ME groups containing EB in mice and the fluorescence intensity and diffusion domain of brain slices were all studied.

RESULTSContrasting to the common microemulsion (ME), HAC-ME at the lower concentration of HAC (<5 mg x mL(-1)) could further improve the transporting of EB across the BBB while EB concentration in other tissues decreased, and Tmax was delayed about 30 min.

CONCLUSIONHAC-ME could facilitate the transporting of EB across the BBB and it was concentration dependent. While the brain targeting absorptive capability of HAC-ME was enhanced.

Animals ; Blood-Brain Barrier ; drug effects ; metabolism ; Brain ; metabolism ; Chitosan ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Emulsions ; Evans Blue ; chemistry ; pharmacokinetics ; Female ; Hyaluronic Acid ; chemistry ; pharmacokinetics ; pharmacology ; Male ; Mice ; Particle Size ; Tissue Distribution

Objective To detect the expression of matrix metallproteinases(MMPs) in aortic valve of children who suffered from rheumatic heart disease(RHD) and to explore the pathological role of MMPs in children′s rheumatic aortic valve disease.Methods RHD group composed of 18 aortic valves from children suffered from RHD.Controls were 8 children who were died accidentally without cardiovascular system diseases.Hematoxylin and eosin stain observing the histological characteristic of the 2 groups.Immunohistochemistry was used to detect expression of MMP2 and MMP9 on aortic valves in 2 groups.Results Hematoxylin and eosin stain showed:in RHD the valves′ structure were destroyed along with fibrous tissue proliferation,mucinous degeneration,collagen and fiber hyalinization,blood vessel and blood capillary proliferation,lymphocyte,plasmocyte,monocyte infiltration.Immunohistochemistry showed that MMP2 and MMP9 expression were significantly higher than those in the aortic of RHD(68.85?13.08,64.35?9.59) compared with control group(107.31?23.39,116.28?6.99)(t=3.92,10.18 all P

AIMTo investigate the mechanisms of action of transportation of liposomes and chitosan-coated liposomes containing leuprolide across rat intestine and Caco-2 cell.

METHODSEverted-gut technique and Caco-2 cell were used to study the transport properties of free leuprolide, liposomes and chitosan-coated liposomes containing leuprolide. Caco-2 cell was used to study the effect of chitosan concentration and the order of addition on the permeation of liposomes.

RESULTSThe transport of leuprolide was passive diffusion. Probably because the entrapment by liposomes prevents the transport of leuprolide across the rat intestine and Caco-2 cell, the permeation amount of leuprolide from liposomes was lower than that of the free drug. However, liposomes protected the leuprolide from degradation. Chitosan promoted the transport of leuprolide from liposomes and there was no obvious difference in enhancement effect from the concentration of 0.1% to 0.5%. On the other hand, the incubation of chitosan with liposomes may weak the enhancement effect of chitosan.

CONCLUSIONChitosan-coated liposomes showed both protection and enhancement effect, therefore, they may promote the oral absorption of leuprolide.

Animals ; Antineoplastic Agents, Hormonal ; administration & dosage ; pharmacokinetics ; Biological Transport ; drug effects ; Caco-2 Cells ; Chitosan ; chemistry ; pharmacology ; Drug Carriers ; Drug Delivery Systems ; Humans ; Jejunum ; metabolism ; Leuprolide ; administration & dosage ; pharmacokinetics ; Liposomes ; Particle Size ; Permeability ; Rats ; Rats, Sprague-Dawley

AIMTo prepare the breviscapine liposomes and study the pharmacokinetics of breviscapine liposomes in Beagle dogs.

METHODSThe cross-over design (two periods) was employed. Six Beagle dogs were administrated a single intravenous dosage of 28 mg of breviscapine liposomes and reference preparation, respectively, scutellarin in plasma of 6 dogs at different sampling time was determined by RP-HPLC. The pharmacokinetic parameters were calculated by 3P97 program and compared by statistic analysis.

RESULTSThe mean concentration-time curves of breviscapine liposomes and reference preparation were both fitted to two-compartment model with the main pharmacokinetic parameters as follows: T 1/2 alpha were (4.4 +/- 0.7) min and (1.8 +/- 1.3) min respectively; T 1/2 beta were (55 +/- 27) min and (28 +/- 23) min respectively; V(c) were (1 580 +/- 265) mL and (2 460 +/- 2 200) mL respectively; CL(s) were (88 +/- 10) mL x min(-1) and (324 +/- 69) mL x min(-1) respectively; and AUC(0-720) were (363 +/- 42) microg x min x mL(-1) and (102 +/- 19) microg x min x mL(-1) respectively. The T 1/2 alpha, CL(s) and AUC(0-720) of breviscapine liposomes all had significant difference from those of reference preparation, after the data were examined by a one-way analysis of variance (ANOVA).

CONCLUSIONCompared with the reference preparation, breviscapine liposomes had a much more higher concentration in plasma and contained characteristic of sustained-release, which ameliorated the pharmacokinetic properties of scutellarin.

Animals ; Apigenin ; blood ; Area Under Curve ; Brain ; metabolism ; Cross-Over Studies ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; Drug Stability ; Erigeron ; chemistry ; Female ; Flavonoids ; administration & dosage ; isolation & purification ; pharmacokinetics ; Glucuronates ; blood ; Injections, Intravenous ; Liposomes ; Male ; Plants, Medicinal ; chemistry

AIMTo evaluate the characteristics, the hypoglycemic efficacy and the pharmacokinetics of the insulin-liposomes double-coated by chitosan (CH) and chitosan-EDTA conjugates (CEC).

METHODSInsulin-liposomes were prepared by reversed-phase evaporation. The protection of insulin against peptic and tryptic digestion was studied with HPLC. The hypoglycemic effects of insulin-liposomes were investigated using the glucose oxidase method after oral administration to rats. Serum insulin concentration in rats were determined by radio-immunoassay, and were assessed by Pkanalyst computer program.

RESULTSThe insulin-liposomes double-coated by CH and CEC was shown to protect insulin against digestion of pepsin, trypsin and gastrointestinal contents. In glucose tolerance test in normal rats, as compared with phosphate buffer solution control group, the insulin-liposomes coated by CH and CEC could reduce the glucose-induced peak of hyperglycemia. The reduction of the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes. When administered intragastrically to normal rats, the insulin-liposomes coated by CH and CEC could reduce glycemia measured after an overnight fast. The hypoglycemic effect the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes, and the dosage of 50 mu x kg(-1) decreased by 45.98% of initial blood glucose level at 1 h. As compared with subcutaneous injection, the relative pharmacological bioavailability was 17.02% calculated by area under the curve of glucose level versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats. The serum insulin concentration-time curves were found to best fit the one-compartment open model. As compared with subcutaneous injection, the relative bioavailability was 8.91% calculated by the area under the curve of serum insulin concentration versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats.

CONCLUSIONThe stability and absorption of insulin-liposomes double-coated by CH and CEC was superior to that of the insulin-liposomes coated either by CH, or by CEC respectively.

Administration, Oral ; Animals ; Biological Availability ; Blood Glucose ; metabolism ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Edetic Acid ; chemistry ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; pharmacology ; Insulin ; administration & dosage ; pharmacokinetics ; pharmacology ; Liposomes ; Male ; Nanotechnology ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Technology, Pharmaceutical ; methods

AIMTo study the effect of the liposomes coated by chitosan and its derivatives as oral dosage form for peptide drugs on the gastrointestinal (GI) transit of drugs.

METHODSInsulin-liposomes were prepared by reversed-phase evaporation. The in situ perfusion experiment was used to investigate the enteral absorption of insulin. The hypoglycemic effects of insulin were investigated using the glucose oxidase method after administration in rats. The insulin concentrations of serum and enteral tissues were determined by radio-immunoassay in rats.

RESULTSIn in situ local intestinal perfusion experiment, the duodenum was the best segment for the absorption of the insulin liposomes coated by chitosan (CH) or chitosan-EDTA conjugates (CEC) , and double-coated by CH-CEC; the colon was the best segment for the absorption of the insulin solution from rat intestine; but the best segment for the absorption of the uncoated and N-trimethyl chitosan chloride (TMC) coated insulin liposomes was unclear. In all segments, the enteral absorption of the insulin liposomes double-coated by CH-CEC was superior to that of other insulin liposomes.

CONCLUSIONThe insulin-liposomes coated by chitosan and its derivatives can enhance enteral absorption of insulin and increase stability of insulin in GI tract.

Animals ; Area Under Curve ; Chitosan ; chemistry ; Colon ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Duodenum ; metabolism ; Edetic Acid ; chemistry ; Gastrointestinal Transit ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; Insulin ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; Liposomes ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relation between the epidemiological strains of meticillin-resistant Staphylococcus aureus (MRSA) and the strains isolated from the nasal fossa of the medical staff and inpatients.

METHODSThe MRSA strains were isolated from the nasal fossa of the medical staff and inpatients in the Department of Neurosurgery. The genes of the isolated strains were amplified by randomly amplified polymorphic DNA (RAPD) assay.

RESULTSThree and 12 MRSA strains were isolated from the nasal fossa of the medical staff and patients who were hospitalized for more than 1 week, respectively, and RAPD assay revealed high homology between the isolated strains.

CONCLUSIONCross infection can be present between the medical staff, inpatients, and the infected patients.

China ; epidemiology ; Cross Infection ; microbiology ; DNA, Bacterial ; genetics ; isolation & purification ; Humans ; Infectious Disease Transmission, Professional-to-Patient ; Inpatients ; Medical Staff ; Methicillin Resistance ; Nasal Cavity ; microbiology ; Phylogeny ; Random Amplified Polymorphic DNA Technique ; Staphylococcal Infections ; epidemiology ; microbiology ; Staphylococcus aureus ; classification ; genetics ; isolation & purification

OBJECTIVETo investigate the dynamic changes of the anti-HBs level among immunized newborn infants born to HBsAg-positive and HBsAg-negative mothers in hyper-endemic area of Hepatitis B.

METHODSInfants who were regularly vaccinated with Hepatitis B vaccine and tested to be anti-HBs positive were divided into two groups according to HBsAg-positive or negative mothers in Long-an, Guangxi. Each subject was followed up 3 times during age 5 to 8. SPRIA was used to test HBsAg, anti-HBs and anti-HBc. Results During the follow-up period, positive rates of anti-HBs in children born to HBsAg-positive mothers ranged between 52.00% and 78.00%, and those with HBsAg-negative mothers was between 43.84% and 54.74%. GMT in two groups was between 55.36 mIU/ml and 95.66 mIU/ml as well as between 39.90 mIU/ml and 65.47 mIU/ml, respectively. There was no statistical significance in both positive rates and GMT between age groups. The anti-HBs level in the follow-up period of children born to HBsAg-positive mothers was higher than that of those born to HBsAg-negative mothers in the same age group. In the age group of 6-8 years with HBsAg-negative mothers, the positive rates in the follow-up period of children with high anti-HBs titers in the primary vaccination were 2.29-2.84 times of those with low titers. The anti-HBs titer of children in a follow-up period was lower than that in the primary vaccination, no matter whether they were born to HBsAg-positive mothers. However, the decline rate of children born to HBsAg-negative mothers was significantly higher than those born to HBsAg-positive mothers (84.91% vs. 61.54%; chi2 = 28.7982, P = 0.000). The incidence rate (25.64%) of a 4-fold increase in antibody titers of children born to HBsAg-positive mothers was significantly higher than that of children born to HBsAg-negative mothers (7.37%) from the primary vaccination to the follow-up period (chi2 = 6.7661, P = 0.009) with was 3.5 times of the latter. Subjects with HBsAg seroconvertion were those with low anti-HBs titers in primary vaccination.

CONCLUSIONThe anti-HBs level decreased slowly in successfully immunized children from age 5 to 8. The chance of natural booster yielded by natural infection increased in immunized children born to HBsAg-positive mothers. The anti-HBs level in the primary vaccination played an important role in prevention of seroconversion of HBsAg.

Child ; Child, Preschool ; Female ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Surface Antigens ; blood ; immunology ; Hepatitis B Vaccines ; immunology ; Humans ; Infant, Newborn ; Male

OBJECTIVETo investigate the relationship between -308 genotype polymorphism in the promoter region of the tumor necrosis factor alpha (TNFalpha) gene and asthenospermia in infertile men.

METHODSAllele-specific polymerase chain reaction (ASPCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to analyze the genotype at position -308 in the promoter region of the TNFalpha gene in 187 infertile male patients, who were divided into Groups A (asthenospermia, n = 60), B (oligoasthenozoospermia, n = 65) and C (infertile patients with normal sperm, n = 62). The levels of TNFalpha in the seminal plasma from these patients were measured by radioimmunoassay, and all the data were statistically analyzed by SPSS16.0.

RESULTSGroups A and B exhibited significant differences from C in the frequency of GA/AA at position 308 in the promoter region of the TNFalpha gene (21.67% and 26.15% versus 8.06%, P < 0.05). Spearman analysis showed a negative correlation between the GA + AA type of the TNFalpha-308 allele and the percentage of grade a + b sperm (r = -0.690, P < 0.05). The level of TNFalpha in the seminal plasma was significantly elevated in Groups A ([4.23 +/- 0.45] ng/ml) and B ([4.29 +/- 0.47] ng/ml) as compared with C ([4.03 +/- 0.66] ng/ml, P < 0.05), but with no significant differences between Groups A and B (P > 0.05). It was also significantly higher in the GA+AA ([4.61 +/- 0.29] ng/ml) than in the GGtype ([4.06 +/- 0.45] ng/ml, P < 0.05).

CONCLUSIONRegardless of sperm density, the frequently of TNFalpha-308 GA/AA is negatively correlated with the percentage of grade a + b sperm, which may be associated with the level of TNFalpha in the seminal plasma. Accordingly, anti-TNFalpha therapy might be effective for asthenospermia, and the measurement of the TNFalpha level in the seminal plasma can be an auxiliary diagnostic marker for male infertility.

Adult ; Alleles ; Asthenozoospermia ; genetics ; Case-Control Studies ; Gene Frequency ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo study isolation, identification and differentiation characteristics of dermal multipotent stem cells from human of different age in vitro culture.

METHODSSkin samples( 1 cm x 1 cm) were harvested from fetus, infant, adult and elderly. The original clones were screened in stem cell medium. The diameter and number of clones were recorded. Analysis of each clone and determination of the expression of various related proteins were carried out.

RESULTSThe number of suspended clones from normal skins of fetus, infant, adult and the elderly were (20. 1 +/-2. 5) x 102 , (15. 8 +/-5. 7) x 102, (10. 8 +/-1.3) x 10(2), (6.2 +/- 1.4) x 10(2), respectively ( P <0.01), while the diameter of the clones from them were (83 +/-12) microm, (55 +/- 10) microm, (46 +/- 12) Lm, (42 +/-8) microm, respectively ( P <0.05). Cloned cells from fetus, infant, adult and elderly could differentiate into neuron cell , neuroglia cell, smooth muscle cell, and adipocyte. The clones from fetus were inclined to differentiate into neuron cells, but those from infant were inclined to differentiate into neuroglia cells, and those from adult and elderly were inclined to differentiate into adipocytes. After 1 month of culture, the clone forming rate of the cells from fetus, infant, adult and elderly were 41. 1% , 25.5% ,17.7% ,15.2% , respectively. The individual clone cells also showed ability of multidirectional differentiation. Nestin, fibronectin, c-Myc, STAT3 and hTERT protein were expressed in all clones.

CONCLUSIONMultipotent stem cells with multi-direction differentiation and proliferation can be efficiently isolated from dermis of human of different age in stem cell culture medium. The number, proliferation and differentiation of dermal multipotent stem cells can be affected by age.

Aborted Fetus ; cytology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Child ; Child, Preschool ; Dermis ; cytology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Multipotent Stem Cells ; cytology ; Pregnancy ; Pregnancy Trimester, Second