Allergens ; immunology ; Animals ; Asthma ; etiology ; Enzyme Activation ; Lung ; pathology ; Matrix Metalloproteinases ; physiology ; Mice ; Respiratory Syncytial Virus Infections ; enzymology ; pathology

Objective To evaluate the precise,accurate and specific of two direct methods for measuring low-density lipoprotein cholesterol(LDL-C)based on the principle of selective hydrolysys. Methods Both DAIICHI and Randox reagents were compared with PVS method and the ultracentrifugally separated HDL and LDL fractions were used.Results Both methods all had good precise,the total CV was 3.96%～4.42%(Daiichi)and 0.78%～3.19%(Randox),repectively.The average concentrations of 48 serum samples were 3.68±1.23 mmol/L(PVS method),3.25±1.11 mmol/L(DAIICHI method)and 3.37±1.21 mmol/L(Randox method),respectively.There was no statistics difference between the results from PVS method and other two direct methods.Furthermore it indicated that the results determined by both direct methods had good relationship with that by PVS method.It also indicated that both direct methods had good specific to LDL-C.The dilute test demonstrated that there were good linearity between both direct methods of LDL-C and the linearity range was 9.28 mmol/L at least.Conclusion The direct methods for determining LDL-C based on the principle of selective hydrolysis possessed good precise and accurate and specific to LDL-C,and was meet with clinical application.

Objective To establish the methods of efficiently isolating chondrocytes from epiphyseal cartilage and observe the basic biological characteristics of the cultured ones. Methods The improved isolation method through three-step enzymatic digestion using 1 g/L trypsase in 1 g/L ethylenediaminetetraacetic acid, 1 g/L hyaluronidase and 2 g/L type Ⅰ collagenase was established to isolate the primary passage chondrocytes from epiphyseal cartilage of young rabbits and its efficiency evaluated. The cellular biological characteristics of the chondrocytes were observed under the inverted and light microscopes. Results (1) All chondrocytes were isolated from the matrix after it was completely digested by three above-mentioned enzymes. The harvested chondrocytes significantly and positively correlated with the wet weight of cartilage. The number of chondrocytes from 1 g of wet cartilage was 32.3?106 with a viability of 97.6% on average. (2) The chondrocytes of the primary and the first passages showed triangle or multi-angle shapes (oval during growth and fusion) with the positive immunohistochemical stain of collagen type Ⅱ and the strong heterochromia to toluidine blue. The chondrocytes after the third passage became spindle-shaped with the negative stain of collagen type Ⅱ and the weak heterochromia to toluidine blue. Conclusions (1) The chondrocyte-isolating method through three-step enzymatic digestion has advantages of high efficiency in harvesting primary chondrocytes, high cellular viability and simple manipulation. (2) The chondrocytes differentiate and lose their special phenotypes gradually with passage.

Objective To develop a new kind of high efficiency and energy efficiency viewbox for medical diagnostic image.Methods By constructing an array of many LED with super bright white light to form viewbox of arrangement reasonable and brightness symmetrical.Results Through all contrast with application in many sorts LED,different permutation and combination and controlling current,its design requirements can be achieved at last.Conclusion The surface-intensity lights and even-degree of the viewbox can satisfy the needs of medical diagnostic image and it is a new kind of high performance and energy-saving viewer.

Objective To establish the technique of biphasic seeding of articular chondrocyte s in vitro for cartilage engineering in u sing a self designed biological gel a nd the three-dimensional scaffold a nd observe the efficiency of cartilage regeneration in vitro-tissue engin eered articular cartilage from cell scaffold complex.Methods The articular chondrocytes were iso lated enzymatically from the epiphyseal cartilage of young rabbits,and were then plated i nto the tissue culture flasks and were cultivated.The first passage ar-ticular chondrocytes were collecte d and mixed fully with the self-made l iquid biological gel-matrix at appr oxi-mately 2.5?10 7 cells /ml to form cell-gel fluid.The cell-gel fluid was dropped into the p orous CPPf /PLLA(calcium polyphosphate fiber /poly -L -lactic acid)scaffold,and a cell-gel-scaffold c omplex was formed after being solidified.The complexes were cultivated for 4weeks.The changes of complexes in morphology and synthesis of collagen typeⅡandⅠand aggregates were investigated under the gross and the phase and light microscope.The GAG sulfate content in complexes was quantitatively mea sured by the modified dimethyl-methylene blue method.Results1)After feeded,the porous CPPf /PLLA s caffolds were completely filled with the liquid chondrocyte-gel and the chondrocyte-gel-scaffold comp lexes were well formed by solidification.During the cultivation,the complex es could keep its original shape and m aintain the stable homogeneous three-dimensional distribution of chondrocytes in themselves without cell falling.In the same time,the com-plexes were gradually increasing th e consistency with the elasticity an d lubrication surface.2)The chondro-genesis began in the periphery area a nd extended to the central area of the complexes with the passage of cultivation period.After the 2nd we ek,the complexes were gradually reorganized into the mature engineered cartilage with typical cartilaginous histological structure,with ric h typeⅡcollagen and the strong typical het-erochromia to toluidine blue,but wi th gradually fading negative immunological stain of collagen typeⅠ.Meantime,the scaffold was graduall y de-graded in the complexes.The average content of GAG sulfate in the enginee red cartilages at 4th week was(15.70?2.00)mg /g(wet weight ),and was over 30%of the natural articular cartila ge.Conclusion The technique of chondrocytes biphasic seeding for three-dimensional scaffold has advantages of simple manipulation,fix ing cell stably in the scaffold,main-taining the original shape of the com plexes and the stable homogeneous th ree-dimensional distribution of chondrocytes in the scaffold,and ad vancing the regeneration and matura tion of tissue-engineered articula r cartilage.[

OBJECTIVETo observe the effect of anisodamine (Ani) injection on the survival rate and histologic result of flaps with ischemia-reperfusion injury, so as to demonstrate the protective effect of Ani on the flap survival.

METHODSA total of 48 healthy male Wistar rats were randomly divided into model control, normal saline(NS) and anisodamine groups, with 16 rats in each group. An 3 cm x 6 cm axial flap was formed at the right lower abdomen with abdominal superficial blood vessel as the pedicle. 0.5 cm x 0.5 cm skin tissue was taken from the middle part of flaps in each group, immediately after operation, 12, 18, 24 h after operation. The superoxide dismutase (SOD), nitric oxide (NO), nuclear factor-kappaB contents in the specimens were detected. The histologic study was also performed. The flap survival rate was recorded 7 days after operation.

RESULTSFlap survival rate was (78.6 +/- 7.3) % in Anisodamine group. 12, 18, 24 h after reperfusion injury, the SOD was (103.3 +/- 3.9), (82.6 +/- 3.8), (67.5 +/- 4.6) U/mg; the NO was (5.33 +/- 2.05), (4.75 +/- 1.68), (4.15 +/-1.59) nmol/mg; the NF-kappaB was 0.211 +/- 0.039, 0.313 +/- 0.033, 0.096 +/- 0.028. The contents of SOD, NO and NF-kappaB had the statistical difference of at different time. The skin pathological changes in Anisodamine group was obviously better than those in NS group. Flap survival rate in Anisodamine group was significantly higher than that in NS group.

CONCLUSIONSIn the flap with ischemia-reperfusion injury, Anisodamine can reduce the damage of free radical, increase the blood flow, reduce the production of NF-KB, decrease inflammatory reaction. So Anisodamine can increase the survival rate of flaps with ischemia reperfusion injury.

Animals ; Graft Survival ; drug effects ; Male ; NF-kappa B ; analysis ; Nitric Oxide ; analysis ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Solanaceous Alkaloids ; therapeutic use ; Superoxide Dismutase ; analysis ; Surgical Flaps ; blood supply ; pathology ; Vasodilator Agents ; therapeutic use

Objective:To investigate the effects of N-acetylcysteine(NAC)on the number of Clara cells and secretion of Clara cell16000(CC16)protein in murine asthmatic model.Methods:The murine asthmatic model was established by sensitiz-ing and challenging BALB/c mice with ovalbumin(OVA).Thirty mice were divided into control,asthmatic and NAC groups (n=10). The number of Clara cells and synthesis of CC16were determined by immunohistochemistry.The CC16level in bronchoalveolar lavage fluid(BALF)was determined by Western blot.Results:The proportions of Clara cells in terminal and respiratory bronchioles were(58.05?3.75)%and(63.70?1.79)%in the asthmatic group,(74.54?5.81)%and (78.46?1.68)% in the control(P

0.05).Conclusion The local relapse rate and the survival rate within 3 years has no significant difference between patients with lower rectal cancer of Duck stage A after sphincter-preserving surgery and Miles surgery.

Whether InfusaSleeve(IS) catheter can deliver antisense oligodeoxynucleotide (ODNs) following arterial denuation is unknown. We evaluate the feasibility of local endoluminal delivery of C-myc ODNs to the site of arterial denudation by using IS catheter and to determine the biological importance of these effects. IS catheter was introduced into right side of iliac artery of 21 rabbits after angioplasty of iliac artery. Animals were randomized to the control group (n＝6) receiving saline injection and the treated group receiving c-myc antisense (n＝15, 1 mg ODNs per vessel). In two weeks and 40 days following the operation, angiography was performed. Morphometric analyses were carried out in balloon-denuded iliac arteries. The expression of c-myc protein was detected by using a mouse monoclonal antibody to c-myc. Morphometric analyses carried out at 40 days after transcatheter c-myc antisense oligomer administration. The results showed that maximal neointimal area was reduced from 7.66±3.7(×105 μm2) in the control group (n＝6) to 4.04±1.02(×105 μm2) in the antisense treated group (n＝6, P＜0.05). These changes in vascular remodeling following denuding injury resulted in an increase in residual luman from 20～50% in the control group to 70～90% in the antisense-treated group. C-myc protein expression was virtually undetectable at baseline in locally ODNs-delivered arteries and detectable in control denuded arteries. The results show that: ①Single IS transcatheter administration allowed endoluminal delivery of ODNs to the site of arterial injury; ② c-myc antisense oligomer reduced the formation of neointime in denuded arteries, implying a therapeutic potential of this approach.