Aim To explore the expression of CD1a＋ DC in fair follicle of normal human skin and psoriasis patients. Methods Expressions of CD1a＋ DC in 10 fair follicles of psoriasis patients and skins of 8 normal human were detected by SABC immunohistochemical staining. Results The numbers of CD1a＋ DC in upper limb skin and scalps's fair follicle were 13 and 15 respectively. The number of CD1a＋ DC in fair follicle of psoriasis patients was 16,slightly more than that in skins. There was obviously higher density of CD1a＋ DC in psoriasis fair follicle and its number was 46± 15/mm2. Conclusion The area of fair follicle is connected with the start and redistribution of CD1a＋ DC,and the fair follicle may have important effect on development of vulgaris psoriasis.

Objective To investigate the protective effects of propofol in clinical relevant concentration on TNF-induced apoptosis in endothelial cells derived from human umbilical vein (HUVECs) .Methods The third and fourth passages of the cultured endothelial cells were divided into 5 groups. 0, 12.5, 25, 50 or 100 ?mol/L propofol was added to the cultured cells respectively and the cells were incubated for 30 min. Then TNF-? was added to the cultured cells (the final concentration of TNF-? was 2000 U.ml-1 ) which was incubated for 24h. Apoptosis was detected by using "terminal deoxynucleotidyl transferase (tdt )-mediated deoxyuridine triphosphate (dUTP )-biotin nick end-labelling" (TUNED . Morphologic changes were examined by means of electron microscope.Results Apoptosis expression was very high in control group (no propofol was added) . The process can be inhibited by pre-treatment with propofol (P

An HPLC-DAD-MS/MS method was developed for rapid analysis and identification of degradation products of buagafuran. Buagafuran and degradation products were separated on a Zorbax C8 column (5 microm, 4.6 mm x 150 mm) using acetonitrile-water (78 : 22) as mobile phase. The elutes were detected with diode array detector and tandem mass spectrometer via electrospray ionization source in positive ion mode. According to analysis of the retention time, UV spectra and MS, MS/MS data, combined with the possible degradation reaction of buagafuran, the structures of main degradation products were inferred. The results showed that six main degradation products were oxidation or peroxidation productions of buagafuran. Degradation product A was a double bond epoxidation product of buagafuran, degradation products B, C, D and E were the further oxidation products of degradation product A, degradation product F was a peroxidation product of buagafuran. The results indicated that the established method was effective in the rapid identification of the degradation products of buagafuran.

BACKGROUND:At present, there is still lack of related reports about the aging process of in vitro cultured epidermal cells, since epidermal cells are seed cells necessary for the construction of tissue engineered skin, this articleis is aimed to investigate the biological property of normal human epidermal cells during aging process so as to provide a foundation for the selection of seed cells for tissue engineered skin OBJECTIVE: To observe the in vitro proliferation and aging property of human epidermal cells in order to provide a foundation for the proper selection of seed cells for tissue engineered skin.DESIGN: A self-comparative experiment.SETTING: Orthopedic Surgery Research Instioute of Weifang Medical College and the General Surgery Department of Weifang Medical College Affiliated Hospital.MATERIALS: This experiment was carried out at the Orthopedic Surgery Research Institute of Weifang Medical College, between September 2000and September 2002. Healthy foreskin tissue was obtained from 20 normal boys of 6-8 years old who received peritomy at the General Surgery Department of Weifang Medical College Affiliated Hospital.METHODS: Epidermal cells were obtained from normal young people for subculture. Cells were collected from different culture passages and taken as subjects, and their aging characteristics were assessed through morphological observation, population doubling time (PDT), immune cytochemistry and beta-galactosidase staining. MAIN OUTCOME MEASURES: ① The changes of the epidermal cell growth characteristics. ② The morphological changes of the epidermal cells. ③ The epidermal cell phenotypic changes. RESULTS: ① The clanges of the epidermal cell growth characteristics: Cells were in vitro cultured by monolayer for 9 passages, and PDT of P2 was the shortest. The cells showed strong proliferation in the first 5 passages.From P6, PDT was obviously prolonged, but the cells from P8 did not proliferate any longer. ② The morphological changes of epidermal cells: The primary cultured cells began to proliferate 3 days later, which accelerated 4 days later. The cells became approximately fused in about 1 week. The growth of epidermal cells was identified with a microscope and the immuno histological techniques. ③ The epidermal cell phenotypic changes: Along with the consecutive subculture, histological expression of beta-galactosidase was found to show an increasing tendency from weak expression (occupying 9% of the young cells) to strong expression (occupying 65% of aging cells), and the positive expression rate of beta-galactosidase was found to be remarkably correlated with cell passage age (r=0.87, P ＜ 0.01). CONCLUSION: ① Compared with young cells, aging cells displayed more obvious aging morphology and enzyme cyto-chemical characteristics.During the cell aging process, the PDT of cells showed an increasing tendency. ②Compared with young cells, the expression of beta-galactosidase in aging cells was remarkably increased, and this increase paralleled with the appearance of cell aging phenotype and the loss of cell proliferation capability, and reflects the aging degree of cells. ③ The in vitro cultured normal human epidermal cell aging model was established in this experiment. The results of this experiment indicated that epidermal cells from the 1st -5th passage (donators aged 16-18 years old) can be taken as the optimal seed cells for tissue engineered skin construction.

BACKGROUND: Fibroblasts are considered as seed cells necessary for the construction of tissue engineering skin. The ultrastructure of cells of various generations was observed under the electron microscope in the hope of providing foundation for proper selection of seed cells for tissue engineering skin.OBJECTIVE: To observe the ultrastructural changes of normal human fibroblasts during in vitro culture.DESIGN: Self-control observation.SETTING: Institute of Plastic Surgery, Weifang Medical College; Department of General Surgery affiliated to Weifang Medical College.MATERIALS: This experiment was carried out in the Institute of Plastic Surgery, Weifang Medical College, between September 2000 and September 2002. Healthy prepuce specimens were collected during posthetomy from normal boys aged 6-8 years after the informed consent was obtained from their guardians.METHODS: The normal human diploid fibroblasts were used to carry out consecutive subculture; cells were collected from different generations for morphological and ultrastructural observation under the inverted phase contrast microscope and transmission electron microscope.ture under the transmission electron microscope.microscope: Cells could pass on for 65-70 generations and survive for 280-300 days. Cells within 45 generations could grow rapidly, but gradually grew slowly after the 45th generation, and even displayed no proliferaunder the transmission electron microscope: There were no obvious changes in cell ultrastructure within 40 generations, but cells presented inward tolds of nucleus membrane from the onset of generations 41-65, with the ratio of cell nuclear/plasma reduced as well as cell surface process and microvilli also reduced.CONCLUSION: The ultrastructural change of in vitro cultured fibroblasts varied between different generations, which became obvious after the 41st generation, suggesting that fibroblasts within 40 generations are considered preferable seed cells for the construction of tissue engineering skin.

Objective To study the role of cutaneous nerve and protease activated receptor 2 (PAR2)in the development of pruritus in atopic dermatitis(AD).Methods Dermal sheets were prepared from chronically pruritic skin lesions of 7 patients with AD,as well as from the normal skin of 7 healthy human controls.Double labeled immunofluorescence was performed using mouse anti-protein gene product 9.5(PGP9.5)monoclonal antibody and rabbit anti-substance P(SP)polyclonal antibody to observe the morphological changes in cutaneous nerve fibers,and Image-Pro Plus 6 software was used to semiquantitively assess the length,diameter of nerve fibers,integral optimal density of PAR2 and SP in dermal sheets.Results Immunofluoresence double staining showed that PAR2 co-expressed with PGP9.5 or SP in cutaneous nerve fibers.Compared with the normal control skin,both the total length and average diameter of PGP 9.5-expressing nerve fibers were increased(11051.8±1900.9 μm vs 7264.0±2659.9 μm,4.23±0.15 μm vs 3.95±0.15 μm,both P＜0.01)in pruritic lesions,while only the average diameter of SP-expressing nerve fibers was up-regulated(3.99±0.20 μm vs 3.80±0.07 μm,P＜0.05),and the total length of them remained unchanged(4304.7±1455.0 μm vs 3380.0±1735.4 μm,P＞0.05).Also,increased integral optimal density was observed for SP and PAR in pruritic lesions in comparison with the normal control skin (27.71±16.52 vs 12.63±4.31.35.99±8.63 vs 22.69±9.56.both P＜0.05).Conclusion Our results indicate a hyper-plasia of cutaneous nerve fibers in chronic itchv skin lesions of AD and an increase in the expression of PAR2 and SP in the cutaneous nerve fibers,suggesting that the signal enhancement in PAR2 pathway may be related to the mechanism of pruritus in patients with AD.

BACKGROUND: As the seed cells for construction of tissue engineered skin, fibroblasts directly decide the quality of tissue-engineered skin. During in vitro culture, collagen gene expression and response to epidermal growth factor (EGF) stimulation of the fibroblasts in different passages can be indicative of their proliferative capability for use as the seed cells for skin tissue engineering.OBJECTIVE: To observe the expression of type Ⅰ and Ⅲ collagen mRNA in fibroblasts cultured in vitro and fibroblast response to EGF stimulation, and thereby providing reference for the selection of optimal seed cells for tissue engineering.DESIGN: Self-controlled experiment.SETTING: Institute of Plastic Surgery, Weifang Medical College.MATERIALS: This experiment was carried out at the Institute of Plastic Surgery, Weifang Medical College between September 2000 and June 2002. The specimens of normal prepuce tissues excised by circumcision were obtained from 20 healthy boys at the age between 6 and 8 years on a voluntarily basis in the Department of General Surgery, Affiliated Hospital of Weifang Medical College.surgically excised prepuce by trypsin and type Ⅰ collagenase digestion. After cultured till 80% confluence, the cells were digested with mixed digescontrast microscope was used for dynamic observation of the cell morphology and growth status, and transmission electron microscopy and anti-vigen gene expression: Reverse transcriptase PCR (RT-PCR) was performed for amplification of type Ⅰ and Ⅲ collagen cDNA derived from the total sis of fibroblast response to EGF stimulation: The fibroblasts of P10 and P60passage were divided into treatment group with stimulation by the conditioned medium containing EGF and control group with treatment with only the conditioned medium. 3H-TdR incorporation assay was performed for analyzing the growth of the fibroblasts in response to EGF stimulation.lasts of different passages to EGF stimulation.decreased with cell passaging and 3H-TdR incorporation was lower in P60cells without significant difference between the treatment group and control group (132.5±23.6 vs 124.9±16.8, P ＞ 0.05) than in P10 cells with,however, significant difference between the two groups (512.8±56.4 vs 306.4±22.5, P ＜ 0.01).EGF stimulation is weaker than P10 cells, moreover additional EGF in the condition medium has no obvious regulation on the proliferation of P60cell growth, but extremely remarkable on P10 cells, implying along with the increase of cell passage, tritium-thymidine incorporation reduced and regulative capability of EGF on aging fibroblastic growth was also attenuated.

Objective To study the dynamic change of polysaccharide from Asarum insigne in Guizhou during different harvest period and its hemostatic effect. Methods Asarum insigne polysaccharide was extracted by water isolation and alcohol precipitation. We measured the polysaccharide content by UV spectrophotometry after impurity and purification and detected the bleeding and clotting time by tail cutting and slide methods in mice. Results There was significant variation in polysaccharide content of Asarum insigne at different harvest time, which was at a higher level in June(1. 78%-1. 82%). The bleeding time in mice of normal control group was (6. 73±1. 21) min,and that in mice treated with refined polysaccharide at high dose was (4. 91±1. 58) min,the difference between two groups was statistically significant(P<0. 01). The clotting time in mice of normal control group was (7. 27±2. 09) min,and that in the refined polysaccharide at middle and high dose groups was (3. 96±1. 78) min and (3. 27±1. 61) min,respectively. The latter two groups were obviously different from the normal control group(P<0. 05 or P<0. 01). Conclusion The polysaccharide is an active hemostatic substance in Asarum insigne and the optimum harvest time of it is in June for the clinical use.

The pharmacokinetics and tissue distributions of the novel paclitaxel microemulsion based on the L-OH lipid complex made in our laboratory were studied in this article with the commercial paclitaxel injection in cremophor as reference preparation by injected intravenously with single dose of 5 mg x kg(-1) in rats. LC-MS/MS method was used to determine the drug concentration in plasma and calculate the pharmacokinetic parameters. [3H]-paclitaxel was used to reveal the tissue distributions of different organs in 0.5 h, 3 h, 24 h and 120 h. The results indicated that the AUC of the emulsion group descended to 42.55%, with the CLz and Vz increased by 2.27 times and 3.81 times respectively. Tissue distribution results revealed that the emulsion showed a significantly increase in liver and spleen with a peak concentration up to 5 times; a slightly increase was observed in lung with no statistical differences; a significantly decrease in heart, kidney, gastrointestinal tract, bone marrow, aorta, thymus, pancreas, fat, muscle, skin, seminal vesicle, reproductive organs and brain with a drop of 40%-80%. These results indicated that paclitaxel microemulsion based on L-OH lipid complexes can remarkably reduced the blood exposure, accelerate plasma clearance rate and increase distribution volume. The fact that paclitaxel microemulsion tended to be uptake by reticuloendothelial system (RES) contributed to the target in liver, spleen and lung, and help to reduce the toxicity in blood, heart, kidney and gastrointestinal tract.

Injectable lipid emulsions have been routinely used in patients since 1960s as a nutritional supplement for patients requiring parenteral nutrition. In recent years, lipid injectable emulsions have been extensively studied as a kind of novel drug carrier, also the quality problems of the lipid emulsion attract more and more attentions gradually. Large diameter tail of injectable lipid emulsions as a significant quality control indicator should pay more attention. Regarding to the defect of detecting large diameter tail of lipid injectable emulsions in our country, the purpose of this article is to summarize the techniques of detecting large diameter tail, illustrate the impacts of large lipid droplet on the quality of lipid injectable emulsions, emphasize the importance of detecting large diameter tail in lipid emulsions and provide guidance for researching and developing lipid emulsions in domestic market.