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 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.[

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 investigate the effect of sustentaculum tali screw placement on outcomes following open reduction and internal fixation (ORIF) of intraarticular calcaneal fractures.Methods A retrospective analysis was made on 139 patients with intraarticular calcaneal fractures treated by ORIF from April 2008 to January 2012.According to Sanders classification,there were 9 patients with type Ⅱ fractures,87 with type Ⅲ fractures and 43 with type Ⅳ fractures.Fracture reduction followed by placement of calcaneal anatomic plate or anatomic locking plate was performed via calcaneal lateral L-shape incision.All the patients received X-rays and CT scans within postoperative 2 weeks.Placement of screws in sustentaculum tail was detected by CT volume rendering to visualize implants combined with multiplanar reconstruction imaging (MPR).Patients were divided into accurate fixation group (Group A,n =28),marginal fixation group (Group B,n =58),and non-fixation group (Group C,n =53) based on radiological results.Intra-and peri-operative parameters,time to partial weight-bearing,time to full weight-bearing,fracture healing time,and clinical outcome were compared among groups.Functional outcome was assessed using American Orthopedic Foot and Ankle Society (AOFAS) Ankle Hindfoot Scale.Results Postoperative X-rays indicated accurate fixation of sustentaculum tail in 64 patients,marginal fixation in 22,and non-fixation in 53,but CT images manifested accurate fixation in 28 patients,marginal fixation in 58,and non-fixation in 53.Group A had operation duration of (93 ±11) min and intraoperative blood loss of (252 ± 27) ml,longer or higher than those in Group B [(85 ±8) min,(194±16) ml] and Group C [(57±6) min,(136 ±13) ml] (P＜O.05).There was no significant difference in mean hospital stay among the three groups (P ＞ 0.05).Ninety-two patients were followed-up for 12-38 months (mean 18.5 months) and 85 patients completed foot and ankle exercise as planned.Moreover,no significant differences were observed among groups concerning time to partial and full weight-bearing,fracture healing time and AOFAS score at the final follow-up (P ＞ 0.05).Conclusion Sustentaculum tali screw placement has no apparent effect on the short-term outcome of surgical treatment for calcaneal fractures.

Objective To assess the clinical application value and advantages of DSCT Angiography on artery injury in Extremities. Methods Seven suspected patients with artery injury in extremities underwent DSCT Angiography, including three cases on artery injury of upper extremity and four cases on artery injury of lower extremity. All the seven patients were performed with DSCT Angiography after the operation. The data of thin slice cross section were reconstructed by MPR, MIP and VR, employing Somatom Definition, German. Contrast-concentration was 350 or 370mg i/ml, total amount was 80-100ml and injection rates was 4.0 ml/s. Results All the images of angiography displayed many clear blood vessels and smooth vessels verge, contrasting with surrounding issues well. They also displayed no ladder artifact or truncation artifact. The image quality was good enough to meet the requirements of clinical diagnosis. Meanwhile the coincidence between preoperative diagnosis and postoperative diagnosis was 100%. Conclusion To suspected patients with artery injury in extremities, DSCT Angiography may be the first choice. Three-dimensional reconstruction angiographys by means of different reconstruction techniques are very helpful in diagnosing and formulating treatment perscription.

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.

Objective To analyze changes of three periodical circulation systems,erythrocyte sedimentation rate and bone marrow cell morphology in children with malaria.Methods The routine tests of hematology by Sysmex KX-21 Counter, erythrocyte sedimentation rate by Westergren method and bone marrow cell morphology were analyzed. Results In 22 cases of malaria the ratio of Hb level below 110 g/L,WBC below 4?10~9/L and PLT below 100?10~9/L was 68.2%, 41.0%, and 77.3%,respectively. The ratio of children with all three parameters (Hb, WBC and PLT) abnormal was 36.4%, with two parameters abnormal was 63.6%. Ninty-five point five percent of malaria children′s erythrocyte sedimentation rate was abnormal. Fifty-nine point one percent of malaria children had hyperplasia anemia bone marrow morphology, 77.3% secondary thrombocytopenia and 54.5% with both of two bone marrow morphology.Conclusions Three periodical circulation systems of malaria children alter notably, especially in PLT and Hb. The majority has erythrocyte sedimentation rate abnormal, and bone marrow cell morphology shows hyperplasia anemia and thrombocytopenia.

Objective To evaluate Roux-en-Y gastric bypass operation on carbohydrate and lipid metabolism in type 2 diabetes mellitus patients with BMI range of 24 -29. Methods Thirty seven cases of type 2 diabetes mellitus patients undergoing Roux-en-Y gastric bypass operation were studied. Body mass index (BMI), glycosylated hemoglobin ( GHbAlc), fasting glucose ( FPG), fasting insulin (FIns) and C-peptide( FC-p), HOMA-IR, oral glucose tolerance (OGTT) including 2 hour insulin (2hIns) and C-peptide (2hC-p) , plasma levels of total cholesterol (TC), triglycerides(TG), high density lipoprotein( HDL-c)and low density lipoprotein ( LDL-c) were measured preoperatively and on 3 months, 6 months, later postoperatively. Result There was no statistically significant difference between BMI values measured preoperatively and postoperatively (P>0. 05 ). Serum levels measured in pre-operative and third and sixth post-operative months were: FPG (8. 8 ± 0. 9, 7. 0 ± 2. 0, 6. 3 ± 0. 6, P<0. 01) ( mmol/L) , GHbAlc (8.2±1.2, 7.0±0.8, 6.2±0.7, P<0.01)(%), FIns(10. 6 ±1. 2, 9.0±0.9, 9.0±0.8, P<0.05)(mU/L), FC-p(1.9±0.5, 1.2 ±0.6, 1.2 ±0.4, P<0. 01) (nmol/L), TG(3.3 ±0.8, 2.7 ±0.9,2.6±0.7, P<0.05)(mmol/L), TC(6.5±1.8, 4.6±0.9, 4.2 + 1.0, P<0. 05) (mmol/L)and LDL-c (3. 6 ±1.2, 2. 8 ±0.8, 2. 7 ±0.2, P<0.01) (mmol/L), 2 hour glucose after OGTT(2hPG) (18. 6 ±3.0, 12.7 ±2.3, 11.4±2.0, P<0. 01) (mmol/L), HOMA-IR(3. 2 ± 1. 7, 2.6±1.6, 2. 5 ±1.3, P<0. 05). Postoperative levels of HDL-c (1. 2 ± 0. 1, 1. 4 ± 0. 4, 1. 4 ± 0. 2, P<0. 01) ( mmol/L) , 2hIns (17. 2 ±3.4, 26. 3 ±4.7, 28. 6 ±4.1, P<0. 01) (mU/L)and 2hC-p(4. 2 ± 1. 0, 6. 3 ± 1. 5, 6. 2 ± 1.4,P<0. 01 ) ( nmol/L) were significantly higher than that of the pre-operative values ( P<0. 01 ).Conclusions Roux-en-Y gastric bypass significantly improves the metabolism of carbohydrate and lipid in type 2 diabetes patients with BMI 24-29, and the effects are not associated with weight loss.

Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.