Objective To investigate the effects of prostaglandin E1 (PGE1) on renal cell apoptosis in rats with diabetic nephropathy(DN). Methods Totally 55 male Wistar rats were intraperitoneally injected with streptozotocin (STZ) to develop DN model.46 successfully established DN rat models were randomly divided into 4 groups:PGE1 group received PGE1 intravenously at dose of 10 μg · kg-1 · d-1 for 10 d (n=12),angiotensin converting enzyme inhibitors(ACEI) group given ACEI orally at dose of 10 mg kg-1·d-1 for 8 W(n=12),PGE1+ACEI group given both PGE1 and ACEI (n=11),DN control group(n=11) and normal control group(n=10) given saline only.All rats were killed after 8 weeks and blood samples or kidney tissue were collected.Blood urea nitrogen (BUN),serum creatine(Scr),albuminuria of 24 h were detected.Renal pathological morphology and apoptosis of renal cells were observed by HE staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) test. Results At 8 week after treatment,the 24-hour urinary albumin levels were decreased significantly in the following order:DN control group＞ PGE1 group ＞ ACEI group ＞PGE1+ ACEI group＞ control group[(374.6±54.1)μg,(570.0±72.5)μg,(253.1±28.9)μg vs.(1123.4±106.2)μg,P＜0.01 or P＜0.05].BUN[(9.3±2.6)mmol/L,(11.0±3.5)mmol/L,(8.4±2.2)mmol/Lvs.(15.1±4.0)mmol/L]and Scr [(74.5±19.2) umol/L,(83.5± 15.8)μmol/L,(64.6±17.3) μmol/L vs.(117.7±33.0)μmol/L]levels after treatment were also reduced in PGE1,ACEI and PGE1 + ACEI groups as compared with DN control group (P＜0.01or P＜ 0.05). Pathological manifestations of all treatment groups showed better results than DN group,and PGE1 + ACEI group was the best.There was no obvious apoptosis in glomerular area with on significant differences between groups.While apoptosis of renal tubules was observed in DN rats. Conclusions Renal tubule but not glomerular cell apoptosis may play some role in Prostaglandin E1 reduciug albuminuria.

Objective To evaluate the performance of parallel test in detecting malaria infection for returned person from malaria endemic area.Methods The blood samples of 484 returnees from malaria endemic area were analyzed and detected by thick blood smear,rapid diagnostic test (RDT) and nest PCR in four companies involving the African labor dispatching.Results The sensitivi ty of thick blood smear and RDT was 0.628 and 0.744 respectively,which of the parallel test was 0.930.On the other hand,the area under the curve (AUC) of parallel test was 0.930 (95％CI:0.895-0.986),which was higher than thick blood smear[0.814 (95％CI:0.724-0.904)]and RDT[0.847 (95％CI:0.769-0.926)].Conclusion Thick blood smear and RDT,which consist of parallel test,could improve the detection sensitivity and accuracy for returnees from malaria epidemical area effectively.This approach is worthy of popularization and application.

Objective To investigate the effects of all trans-retinoic acid (ATRA) on the expressions of iron metabolism-related genes and their products in K562 cells and the possible relationship. Methods (1) The characteristics of K562 leukemic cell differentiation induced by ATRA was evaluated by Benzidine, Wright's, NSE and NBT staining.(2) The expression levels of cellular surface antigens (CD71 and CD 13) in K562 cells cultured with ATRA were measured by flow cytometry. (3) IRP/IRE binding activity was assessed by RNA/protein band-shift assay.(4) Ferritin was determined by radioimmunoassay.(5) The mRNA expression levels of H-Fn, TfR and IRP2 in K562 cells cultured with different concentrations of ATRA were delineated by RT-PCR method, confirmed by sequencing of RT-PCR products. Results K562 cells could be induced to differentiate into neutrophils by ATRA, confirmed by cytochemical staining. The expression of CD71 decreased while CD13 increased. The mRNA expression levels of TfR and IRP2 were decreased while mRNA expression level of H-Fn was increased in K562 cells cultured with ATRA, compared to that in control cells. Concomitantly,IRP binding activity was significantly decreased but the level of ferritin was significantly increased in K562 cells cultured with ATRA. Conclusions During the course of K562 cells induction and differentiation to myelocytes by ATRA, the expression level of iron metabolism-related genes and products were changed but the upstream-regulation mechanism still remains unclear.

Objective To investigate the effects of heparanase and E-cadherin on the invasion and metastasis of gastric cancer. Methods Fifty specimens of gastric cancer which had been resected at Cancer Hospital of Liaoning Province from February 2005 to May 2007 were collected. The expression of heparanase mRNA and E-cadherin mRNA in these gastric cancer specimens was detected by RT-PCR, and the expression of E-cadherin in these gastric cancer specimens was detected by immunohistochemistry. Data were analyzed by t-test and variance analysis, and the enumeration data analyzed by chi-square test. Results There were significant differences in the expression of heparanase and E-cadherin between gastric cancer cells with high and low differentiation, presence and absence of metastasis, and TNM stages Ⅰ and Ⅱ versus Ⅲ and Ⅳ (t = 1.999, 4.258, 1.735 ; 1.286, 6.794, 3.091; χ~2 =6.273, 9.397, 5.640, P <0.05). The co-expression of heparanase (+) and E-cadherin (-) was correlated with tumor undifferentiation, lymph node metastasis and advanced TNM staging (χ~2 =11.306, 10.208, 8.420, P <0.05). Conclusions Heparanasc shows high expression while E-cadherin shows low expression in gastric cancer tissue. There is a synergistic effect between the abnormal expression of heparanase and E-cadherin, and the gastric cancer cells with coexpression of heparanase and E-cadherin have more malignant potential.

BACKGROUND: Proliferation and differentiation of mesenchymal stem cells (MSCs) is associated with platelet concentration in platelet-rich plasma (PRP). Low enrich multiple cannot reach proper effects, but high level had inhibitory effects on osteoanagenesis.OBJECTIVE: To observe the effect of different volume fraction of PRP on dog BMSC proliferation.DESIGN: A cytological in vitro study.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: Dog BMSCs of 5 passage were adjusted to 3×10~8/L, and incubated in a 96-well plate at 200 μL per well. Following 24 hours of routine culture, primary medium and non-adherent cells were discarded. Prepared PRP gel was mixed with serum-free low-glucose DMEM containing penicillin and streptomycin, and then diluted into 5%, 6.25%, 7.5%, 8.75%, 10% volume fraction. 200 μL above-described liquid was added into the 96-well plate, which was subsequently placed in a incubator.We set up a blank control.MAIN OUTCOME MEASURES: MTT was used to investigate effect of different volume fraction of PRP on dog BMSC proliferation.RESULTS: Compared with the blank control group, various volume fraction of PLP could promote dog BMSC proliferation in early stage. With prolonged time, proliferation speed began to increase at day 6 in the 8.75% and 10% PRP groups, entering platform stage. BMSC number was increased rapidly in the 5% and 6.25% PRP groups, especially in the 6.25% PRP group.CONCLUSION: PRP gel could promote BMSC proliferation markedly and proliferation strength of BMSCs was correlated to the density of PRP. BMSC proliferation would be accelerated by the low density of PRP.

BACKGROUND: Platelet-rich plasma (PRP) contains abundant growth factors that were needed for osteanagenesis. Moreover,the proportion of each growth factor formed by an organism, with good synergism.OBJECTIVE: To explore the influence of PRP on osteogenetic activity of canine bone marrow mesenchymal stem cells (BMSCs) after induction in vitro.DESIGN, TIME AND SETTING: The in vitro cytological experiment was performed at the Central Laboratory of Xiangya Hospital of Central South University from June 2007 to February 2008.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: The 3~(rd) generation BMSCs were collected and divided into 4 groups. BMSCs in the control group were incubated in standard medium. BMSCs in the osteogenetic induction medium group were incubated in high-glucose DMEM containing fetal calf serum, dexamethasone, beta-sodium glycerophosphate and vitamin C. BMSCs in the PRP group were incubated in low-glucose DMEM containing 6.25% PRP. BMSCs in the combination group were incubated in high-glucose DMEM containing 6.25% PRP, dexamethasone, beta-sodium glycerophosphate, and vitamin C.MAIN OUTCOME MEASURES: Alkaline phosphatase activities were measured. Expression of collagen type I was examined by immunocytochemical staining. Calcium tuberoses were labeled using modified Von Kossa staining. Expression of osteocalcin mRNA was examined by RT-PCR.RESULTS: Levels of alkaline phosphates of all groups became increased along with time. The alkaline phosphates level of combination group was strongest (P < 0.05). Following 7 and 14 days of induction, type I collagen expressed positively in the osteogenetic induction medium and combination groups, but negatively in the PRP and control groups. Following 14 days,formation of calcium nodules were found in the osteogenetic induction medium and combination groups. Following 7 and 14 days,expression of osteocalcin mRNA were similar between the control and PRP groups (P > 0.05), which was significantly lower than the osteogenetic induction medium and combination groups (P < 0.05). Expression of osteocalcin mRNA was significantly lower in the osteogenetic induction medium group compared with the combination group (P < 0.05).CONCLUSION: PRP gel can effectively promote osteoblastic effect of BMSCs after induction in vitro following induction in osteogenetic medium.

Objective To explore the operative method of open distal tibial pilon fractures, and to evaluate the outcome of ankle joint function postoperatively. Methods From March 2003 to March 2007, 24 patients with open Pilon fractures were treated with one-stage open reduction and internal fixation (18 males and 6 females) . The average age was 37. 6 years (14 ～ 53 years) . All 24 patients had open fracture, 12 of whom combined fibular fracture. According to AO comprehensive classification system, the fractures was classified as C1 in 4, C2 in 9, and C3 in 11. According to Gustilo-Anderson classification method, the fracture was classified as Type Ⅰ in 3, Type Ⅱ in 5, Type Ⅲ A in 4, Type Ⅲ B in 10, and Type Ⅲ C in 2. All tibial pilon fractures were treated by radical debridement, one - stage open reduction and internal fixation. Soft tissue defection was covered by a vascularized flap and continually washed by pipes under the flap. Results All patients were followed-up at an average of 2. 3 years(1～3.8 years) after the surgery. All the fractures healed at an average of 22. 3 weeks (16 ～54 wk) postoperatively. According to the scoring system of Con-roy, 17 were excellent (62. 5%), 4 good (25%), and 3 poor (12.5%), The excellence rate was 87.5% . According to the ankle score of Teeny and Wiss, there were 11 excellent (37. 5%), 7 good (37.5%), 3 fair (16.7%), and 3 poor (8.3%) and the excellence rate was 75% . Conclusion One-stage management for open Pilon fracture has the advantages of fewer complications, lower infectious rate, and better ankle joint function.

Objective To explore the effect of ACE-inhibitor perindopril on the expression of scavenger receptor A (SR-A) gene in the kidney of diabetic rats.Methods Diabetes were induced in male Sprague-Dawley rats by peritoneal injection with streptozotocin (60mg/kg).The rats were then random di vided into normal control group, diabetes group and ACEI treatment group [4mg/(kg·d) for 24 weeks].Blood glucose concentration and 24h urinary albumin excretion were determined.The renal morphological change was observed.Immunohistochemistry was used to analyze CD68 positive macrophages,and the Mrna of SR-A in renal tissue was detected by quantitative real-time PCR.Results Compared with normal control group,blood glucose concentration,24h urinary albumin excretion and the number of CD68 positive macrophages were significantly increased [(5.3 ± 0.6) mmol/L vs (26.7 ± 3.3) mmol/L;(2.7 ± 1.3) mg/24h vs (26.7 ± 1.8)mg/24h;(0.77 ±0.24)/gcs vs (2.55 ±0.46)/gcs;(6.13 ±0.50)/HPF vs (11.9 ±2.12)/HPF;P ＜0.05],and the expression of SR-A Mrna were significantly up-regulated in diabetes group [ (5.6 ± 1.2 vs 1.5 ±0.2),P ＜0.05].After intervention with ACE-inhibitor,the up-regulations of the above mentioned parameters,except blood glucose concentration,were all significantly inhibited [ (3.6 ±1.4)mg/24h;(1.03±0.37)/gcs;(8.28±1.19)/HPF;3.4±0.7;P ＜0.05].Conclusion ACE-inhibitor might have renoprotective effects of diabetic nephropathy,it probably was associated with inhibiting the expression of SR-A gene.

Objective To construct a detailed, 3-dimensional, anatomically accurate finite element (FE) model of lumbar L4-L5 segment from CT data with a new kind of computer aided design (CAD) method. Methods A modified "no-seed region segmentation" was done to extract the interest region in the CT scan images and produce a binary image. "Best cross-section planes" accounting for the preferential direction dictated by lumbar spine were placed on the initial iso-surface model, forming a "non-regular piecewise subspace". This subspace and the embedded iso-surface mode were transformed by local affine transforms to a "regular subspace", in which a surface mesh of high quality was generated quickly. Finally a reverse transform procedure was employed to recover the shape feature of the lumbar surface mesh of lumbar L4-L5 in the original 3-dimensional space, which was then imported into ANSYS for the 3-dimensional FE mesh construction. Results All complicated anatomical features of the L4-L5 segment were explicitly represented in the unprecedented finite element model. The predicted results for compression, flexion and extension correlated well with experimental data under similar loading configurations. Conclusion The presented CAD method containing advanced algorithm implements fast and accurate simulation of such complicated geometry with fine mesh representation for lumbar FE analysis.

Objective The purpose of this study was to evaluate the protective potential of the Aspergillus fumigatus thiore -doxin reductase GliT ( TR) antigen by establishing and optimizing ELISPOT assay for TR antigen-specific T cells ( TR/AST) secreting IFN-γand IL-4 in peripheral blood mononuclear cells ( PBMCs ) and explore the role of TR/AST in invasive aspergillosis ( IA ) . Methods We optimized the reaction conditions of ELISPOT by preliminary checkerboard titration and determined the frequencies of positive spot-forming cells ( SFCs) specifically secreting IFN-γand IL-4 in the PBMCs of 20 healthy individuals with TR as specific stimulant and with PHA and PMA as positive controls ,. Results Checkerboard titration demonstrated the best result of ELISPOT with the TR antigen at the final concentration of 10μg/well and PBMCs at 3 ×105/well.The median frequency of IFN-γSFCs was sig-nificantly higher (15 [3.5, 59.5]) than that of IL-4 SFCs (0 [0, 0]) (P<0.001).TR induced IFN-γresponses in all the 20 healthy donors, including 9 cases of strong IFN-γresponse (SFCs>20/3 ×105 PBMCs), accounting for 45%, but failed to induce IL-4 response in 19 of the healthy individuals . Conclusion The Aspergillus fumigatus TR antigen could induce an immunodominant Th1 response , and therefore might be a potential protective antigen .