BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results of BMSCs/ HAB composite. RESULTS: ① Iliac bone block cells were cleaned with good reservation of bone matrix. ② The level of ALP and OCN of MSCs was higher after in ducing for 8 days than that in control group [MSCs after induction: (181.54±40.01) nkat/L, (7.2±1.3) μg/L. There was no method to detect the level in control group, P ＜ 0.05]. ③ BMSCs were adhered and grew well in HAB scaffold. CONCLUSION: HAB loaded with induced BMSCs has an excellent os teogenic function in vitro and shows an effective potential as a good bone tissue engineering material.

Objective To investigate the role of heat shock poteins(HSPs)in the pathogenesis of psoriasis.Methods Expression of HSP27,HSP70and HSP60was detected by immunohistochemical and image analysis in epidermal keratinocytes of pre-and post-treatment psoriatic lesional,and non-lesional skin in25psoriatic patients and6healthy controls.Results There was constitutive expression of HSP27and HSP70in epidermal keratinocytes of psoriatic non-lesional skin and normal controls,and the weak expression in psoriatic lesions.There was expression of HSP60in keratinocytes of psoriatic skin,but no expression of HSP60in non-lesional and normal skin.Expression of HSP27and HSP70recovered gradually in post-treatment psoriatic lesions,and no expression of HSP60was observed in lesional skin after treatment.Conclusion HSPs may play a certain role in psoriatic stress protective mechanism.

Objective To investigate the effects of pituitary adenylate cyclase activating polypeptide (PACAP) and its receptor (PACAP-R) on the pathogenesis of psoriasis. Methods The expression of PACAP and PACAP-R in the skin from 10 normal controls, 25 psoriatic lesions and non-lesional skins was measured by immunohistochemical technique. Results The expression of PACAP and PACAP-R was significantly lower in the psoriatic lesional skins than that of the non-lesional skins. The area density and mean absorbance of PACAP and PACAP-R in the lesional skins were significantly lower compared with those in the non-lesional skins (P

Objective Little research has been done on how Cx37 changes the current density of mononuclear macrophage in atherosclerosis.The purpose of this study was to detect the effects of Cx37 on the current density of mononuclear macrophage in atherosclerosis.Methods A total of 30 Wistar mice were randomly divided into Cx37+ group and Cx37-group equally.The atherosclerosis model was constructed by a high-fat diet.According to different parts of sample collection, these two groups were subdivided into Cx37+ plaque group, Cx37-plaque group, Cx37+blood group and Cx37-blood group.RT-PCR was applied to detect the expression of Cx37 in different body parts.The mononuclear macrophages were cultured after being separated from blood and plague in both groups.The current density of mononuclear macrophage was detected by the whole cell recording.Results The relative expression of Cx37 in Cx37 + plaque group was higher than that in plaque group ([1.10±0.02] vs [0.60±0.03]).Energy Spectrum CT was used to detect the carotid artery plaque in both Cx37 + and Cx37-groups, which verified the successful model construction.At 80,120 and 160ms, the current density in Cx37 + plaque group([0.61± 0.06], [0.67±0.07], [0.91±0.03]A/cm2) was significantly higher than those in Cx37 + blood group([0.49±0.02], [0.61±0.03], [0.67±0.02]A/cm2) , Cx37-plaque group([0.48±0.02], [0.60±0.02], [0.64±0.02]A/cm2) and Cx37-blood group([0.49±0.02], [0.59±0.02], [0.64±0.02]A/cm2).The same goes for those at 200, 240, 320ms(P<0.05).Conclusion Cx37 has more significant impact on the current density in the plaque of mononuclear macrophage than in the peripheral blood in promoting macrophages activation and atherosclerosis progress.

Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 μmol·L(-1)) on angiotensin II (Ang II, 0.1 μmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.
Angiotensin II ; metabolism ; Antioxidants ; pharmacology ; Carotenoids ; pharmacology ; Cell Adhesion ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Monocytes ; cytology ; NF-kappa B ; metabolism ; Reactive Oxygen Species ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.

Objective It aims to investigate the relationships among the categories of Comprehensive Version for Stroke as described in the International Classiifcation of Functioning, Disability and Health (ICF) Core Set, and to provide new supports for Judicial Appraisal of functioning in stroke by ICF functioning mapping.Methods The variables of 59 categories of ICF assessment scale and the samples of 106 persons’ are selected and used in the least absolute shrinkage and selection operator (LASSO) for mining dependencies among those variables. The graphical modeling and analyzing with the software Gephi provides a visual map of the correlations among those classiifcations. Results 59 interconnected categories which organized into the functioning mapping. b340, b735, b175 and b152 are centrally positioned categories because of their high correlation.Conclusion Functioning mapping by graphical modeling can reveal complex relational structures embedded in functioning classiifcations, which provides the support for using ICF to appraisal stroke.

Objective:To explore the effect of adipose derived mesenchymal stem cells to regulate the differentiation of macrophage RAW264.7.Methods:First,we used RAW264.7 cells to simulate macrophage and induced them to M 1 macrophage with lipopolysaccharide ( LPS,1 μg/ml) .Then we cultured these RAW264.7 cells in culture mediums which were previously used to culture adipose derived mesenchymal stem cells to imitate the transplantation of ADMSC .Last,the mRNA relative expression of IL-10, IGF-1,Arg-1,TNF-α,FIZZ1,SPHK-1 was detected by real-time PCR.The protein expression of IL-12 p40,IL-27 Rα,IL-10 was detected by Western blot.Results:After been cultured in ADMSCCM and induced by LPS ,M1 markers (TNF-αmRNA,IL-12 p40;P<0.05) of the RAW264.7 cells declined while M2 markers (IGF-1 mRNA,IL-10 mRNA,IL-10;P<0.05) rose.Conclusion: ADMSC can secrete soluble cytokines to induce the RAW264.7 cell,which have been induced to the M1 macrophages,to differentiate towards M2 macrophages.

OBJECTIVETo clarify drug properties of components in Euodiae Fructus.

METHODThe rat cold syndrome model was induced by cold water stress method. The content of neurotransmitters sand hormones such as DA, 5-HT, NE, AChE and 17-OHCS in serum of model rats were taken as the indexes to evaluate drug properties of components in Euodiae Fructus.

RESULTEuodiae Fructus and its components could correct or relief the content of energy metabolism and substance metabolism-related neurotransmitters sand hormones in serum of model rats with water-stressed cold syndrome.

CONCLUSIONEuodiae Fructus and its components are proved to show hot property.

Animals ; Cold Temperature ; adverse effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gastric Mucosa ; drug effects ; pathology ; Male ; Medicine, Chinese Traditional ; Neurotransmitter Agents ; blood ; Rats ; Rats, Wistar ; Stomach Ulcer ; blood ; drug therapy ; physiopathology ; Stress, Physiological