Structures Characteristics and Bioactivity of Polysaccharide CALB-2 from Aurantii Fructus / 中国实验方剂学杂志

Objective:To isolate and purify a polysaccharide CALB-2 fraction from Aurantii Fructus，and analyze its basic chemical structure, morphological characteristics and bioactivity. Method:A refined CALB-2 was obtained from Aurantii Fructus by hot water extraction，then separated and purified by ion exchange resin，ion exchange agarose gel and propylene dextran gel to obtain homogeneous polysaccharide CALB-2. The molecular mass of CALB-2 was determined by high performance liquid chromatography （HPLC）. Monosaccharide composition analysis of CALB-2 was conducted by methylation analysis and Smith degradation. Structural analysis and morphological characterization were conducted by infrared scanning （IR） and scanning electron microscopy （SEM） analysis. Antioxidant activity of CALB-2 was studied by using H2O2-induced cardiomyocyte oxidative damage model. Result:CALB-2 was a homogeneous polysaccharide and the molecular weight of CALB-2 was estimated to be 3.57×107 Da，which was proved to be a kind of highly branched acidic polysaccharides in IR analysis, methylation analysis and Smith degradation, mainly present in form of 1→3，4 bonds. Through SEM observations，we indicated that the molecular morphology of CALB-2 was amorphous solid. The in vitro activity test showed that CALB-2 had obvious protective effects on injury of H9c2 myocardial cells induced by H2O2. Conclusion:CALB-2 is a kind of homogeneous polysaccharide extracted from Aurantii Fructus， with an anti-cardiomyocyte oxidative damage effect, laying a theoretical foundation for further study of Aurantii Fructus polysaccharides．

Study on anti-radiation effect of Citrus aurantium L. polysaccharides-B / 中国药理学通报

Aim To investigate the protective effect of Citrus aurantium L. polysaccharides-B(CALB) on ra-diation induced by 60Co γ-ray in mice. Methods The BALB/c mice were randomly divided into blank control group, radiation model group, CALB administration group (high, medium and low dose), and positive control group(black fungus polysaccharide,HP). The mice were administered orally for 30 days. After the last administration for three hours,the survival rates on the 2nd day and the 14th day of the blank control group and the irradiated mice after the single radioac-tive irradiation (7 Gy) with 60Co γ-ray were meas-ured. In addition, DNA content and micronucleus of bone marrow cells, SOD, GSH-Px activities, MDA content in serum, liver and brain tissues in mice, TChE activity in brain tissues and spleen and thymus index of mice were detected after one-time whole body irradiation with 60Co γ-ray (3 Gy). Results Each dose group of CALB could significantly improve the survival rate of irradiated mice,increase the DNA con-tent of mouse bone marrow cells and reduce the number of micronuclei in bone marrow cells. In addition, CALB could also increase the thymus and spleen index and the levels of SOD and GSH-Px in serum,brain and liver tissues of mice,and reduce the content of MDA. Conclusion CALB has protective effect on radiation injury,which can be used for further development and utilization of Fructus aurantii.

Ectopic expression of keratin 19 and integrin beta1 during wound healing process after microskin grafting in rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the characteristics of keratin 19 and integrin beta1 expressions in the wound after microskin grafting , and to investigate the healing mechanism.</p><p><b>METHODS</b>Full layer skin defects were created in twenty Sprague-Dawley rats and they were divided into two groups, i.e., A group (with grafting of autologous microskin accounting 10% in weight of epidermis loss from skin defect), B group (with grafting of autologous microskin and allogeneic microskin, accounting 10% and 40% weight of epidermis loss respectively in skin defect). The wound healing rate and contraction rate were observed at 2,3,4 post-grafting week (PGW), and the expression and distribution of keratin 19 and integrin beta1 were observed at 2 and 4 PGW.</p><p><b>RESULTS</b>The wound healing rate in the B group on 2 and 3 PGW was obviously higher than that in A group [(85 +/- 5)% vs. (53 +/- 10)%, (84 +/- 8)% vs. (65 +/- 9)%, P < 0.01]. No obvious difference in wound contraction rate between the two groups was observed on the 2, 3 and 4 PGW (P > 0.05). Cells with expression of keratin 19 and integrin beta1 were observed in the suprabasal layers of the epidermis in healing wound, but not in the basal membrane. Integrin beta1 positive expression cells were not observed in the suprabasal layers until 4 PGW.</p><p><b>CONCLUSION</b>Mixed grafting with autogenous and allogenous microskin can improve wound healing. Ectopic expression of keratin 19 and integrin beta1 exists during wound healing process after microskin grafting.</p>

The Procaryotic Expression, Purification and Activity Analysis of VIP-sTNFRII / 中国生物工程杂志

A prokaryotic expression plasmid containing VIP (vasoactive intestinal peptide) and sTNFRII(soluble tumor necrosis factor receptor II ) genes was constructed. The sTNFRII was cloned by PCR by using special primers which contained VIP gene ORF and a linker in its forward primer. The amplified fragment was inserted into the expression vector pET32a between BamHI and Hind III restriction sites. Transformed E.coli DH5 by pET32a-VIP- sTNFRIIexpressed the fusion protein. After being identified, the protein was purified by ion exchange chromatography and by hydrophobic interaction chromatography. The reconstructed protein showed high bio-activity and could be applied for further use.

Influence of mixed grafting of autologous and allogeneic microskin on burn wound healing in rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the influence of mixed grafting of autologous and allogeneic microskin on burn wound healing.</p><p><b>METHODS</b>Autologous microskin grafting (expansion rate 5:1) was employed as control. Autologous microskin mixed with the allogeneic microskin with the thickness of 0.3 mm and 0.6 mm, respectively, were designated as experimental groups 1 and 2 (EP1 and EP2). The wound healing rate, wound contraction rate, and histological changes were observed on the 2nd, 3rd and 4th weeks after the grafting.</p><p><b>RESULTS</b>The wound healing rate in two experimental groups (94.58 +/- 3.99)% in EP1, and (95.28 +/- 1.93)% in EP2 was significantly higher than that in the control group (88.28 +/- 6.85)% at the end of the 2nd week after the grafting (P < 0.05) The wound healing rate in experimental group 2 (94.55 +/- 3.47)% was obviously higher than that in control (88.51 +/- 5.59)% and experimental group 1 (89.51 +/- 4.70)% at the end of the 3rd week after grafting (P < 0.05). There was no obvious difference in wound healing rate among the three groups at the end of the 4th week after grafting. Obvious lymphocytic infiltration was observed by histological examination between epidermis and dermis in the two experimental groups at the end of the 2nd week after grafting. But there was no obvious difference among the three groups 4 weeks after grafting.</p><p><b>CONCLUSION</b>The wound healing could be improved by mixed skin grafting with appropriate quantity of allogeneic and autologous microskin. Furthermore, the wound contraction could be ameliorated if the thickness of allogeneic dermis was increased in the mixed grafting even with the same proportion of allogeneic to autologous microskin.</p>

The surveillance and molecular epidemiology of Aspergillus in the environment and patients at liver transplant department and intensive care units / 中华皮肤科杂志

Objective To explore the possible relationship between environmental contamination by Aspergillus and invasive aspergillosis.Methods From November 2005 to October 2006,samples were collected from the environment (air in corridors,air in wards,surfaces and tap water) twice a month,and from patients (nose,pharynx and sputum) at a liver transplantation department (LTD),neurologic surgery intensive care unit (NSICU) and central intensive care unit (CICU) in our hospital,and subjected to fungal culture.The Aspergillus density was determined in these environments.The isolates of Aspergillus flavus were genotyped by random amplification of polymorphic DNA (RAPD) assay to investigate the origin of infection.Results The mean aspergillus density was 12,10.75,0 and 20 cfu/m~3 at LTD,NSICU,CICU and corridors respectively.The five most prevalent species of aspergillus in these environments in decreasing order were Aspergillus flavus,Aspergillus fumigatus,Aspergillus niger,Aspergillus versicolor and Aspergillus clavatus.RAPD demonstrated that the genotypes ofA.flavus isolated from two patients were identical to those of the environmental strains in NSICU.The A.flavus genotypes from 3 patients in CICU were all different from those of the environment strains in CICU,but the genotypes were identical from two of the three patients.Conclusions Aspergillus contamination of different degree does exist at LTD,NSICU and CICU. The genotypes of A.flavus are identical from patients and environment in NSICU,suggesting that the clinical infection may originate from hospital environment.