Objective To investigate the effect of simvastatin on the mobilization and migration of endothelial progenitor cells(EPCs).Methods EPCs were harvested from the bone marrows of two rabbits,cultured with M199,and identified by immunohistochemistry.The identified EPCs were then treated with simvastatin with different concentrations(0,0.01,0.1,1.0 ?mol/L),and their migration induced by simvastatin was determined with Transwell chamber assay.Six rabbits models of cranial bone defect were established and divided into control and experiment groups(3 in each).In order to elicit the effects of simvastatin on mobilization of EPCs,simvastatin was embedded in polylactic acid compound,and implanted into the cranial bone defect area in the experiment group.Meanwhile,polylactic acid was implanted in the control animals.After 10 days,the expression rate of CD34+/CD133+ EPCs in the rabbit peripheral blood was counted by flow cytometry to determine the motivating effect of simvastatin.Results In Transwell experiment,16 hours after adding simvastatin(0,0.01,0.1 or 1.0 ?mol/L),the cell migration ability was obviously increased showing a dose-dependent trend(OD value:0.077?0.014 in control group and 0.075?0.013 in 0.01 ?mol/L group vs 0.097?0.011 in 0.1 ?mol/L group and 0.099?0.019 in 1.0 ?mol/L group,P

Objective To investigate the effect of simvastatin-polylactic acid compound on critical calvarial defects in rats.Methods Twenty male SD rats(150 g?10 g),were used to establish critical cranial defect(10 mm in diameter)model.The animals were randomly divided into control and experiment groups(10 in each).In the control group,40 mg of polylactic acid were implanted into the defect area;whereas in the experiment group,simvastatin-polylactic acid compound were used(20 mg simvastatin and 40 mg polylactic acid).Four and eight weeks after the implantation,the defect area of the rats was observed by X-ray and toluidine blue staining.Results Eight weeks after the operation,X-ray examination showed high-density regions in the defect area in the experiment group,while low-density regions in the control group.The radiopacity of cranial defects were 27.33%?2.54% in the control group,and 74.63%?2.42% in the experimental group(n=5,t=-30.148,P=0.000).Toluidine blue staining showed a few new bone tissues at 4 weeks and fully filled bone defect at 8 weeks in the experiment group.Meanwhile,in the control group,only a small quantity of new bone tissue could be seen on the edge of the cranial defects.Conclusion Locally implanted simvastain-polylactic acid compound is a promising method for the treatment of bone defect owing to its high osteogenic ability.

ve To compare the effects of coronary-artery-bypass (CAB) surgery with and without cardiopulmonary bypass(CPB) on hemodynamics and the function of the grafts. Methods Thirty-five patients undergoing elective CAB surgery were studied. CAB was performed either with hypothermic CPB (n = 15) or without CPB(off-pump, n = 20) . The patients were premedicated with intramuscular pethidine 50 mg and scopolamine O.3mg. Anesthesia was induced with intravenous midazolam 5-15mg, fentanyl 5-20?g?kg and pancuronium 0.1 mg?kg-1 or pipecuronium 0.1 mg?kg-1 and maintained with iv infusion of fentanyl 6-10?g?kg-1, propofol 2-4mg?kg-1?h-1 and intermittent boluses of pancuronium, midazolam supplemented with 1%-1.5% isoflurane inhalation. In off-pump group naso-pharyngeal T?was maintained at 37.2℃ during operation. The amount of heparin used was equal to about one-third of amount used during CPB and ACT was maintained above 250 seconds. MAP was maintained at 70-90 mm Hg. While blood vessel was being grafted onto the coronary arteries heart rate was maintained at 60-80 bpm, otherwise esmolol 10-20mg was given iv every 5 min until it was satisfactorily controlled. In CPB group, during CPB naso-pharyngeal T?was maintained at 32℃-34℃, MAP at 50-70 mm Hg and blood gases and electrolytes within normal range. Right radial artery was cannulated and 7.5F Swan-Ganz catheter was inserted via internal jugular vein into pulmonary artery for hemodynamic monitoring and blood gas analysis. ECG, SpO2 were continuously monitored during operation. At the end of operation in patients with internal mammary artery used as graft, the flow rate of grafts was measured with 3mm Doppler probe.Results (1) After CAB cardiac index (CT) increased significantly in off-pump group(P0.05) . (3) There was no significant difference in the blood flow of artery graft and myocardial oxygen delivery (MDO2), and consumption ( MVO2) as well as MDO2/MVO2 between the two groups. Conclusions Off-pump CAB surgery has less effects on hemodynamics but systemic and myocardial oxygen delivery and consumption are similar between the two groups.

AIM: To established an HPLC/MS/MS method for the study of pharmcokinetics of PMEA-Na (the mono-sodium salts of 9-[2-(phosphonomethoxy) ethyl] adenine) in beagle dogs. METHODS: PMEA-Na and internal standard 9-(3-phosphony-methoxypropyl) adenine were isolated from plasma by protein precipitation with methanol, and then analyzed adopting multiple reaction monitoring (MRM) mode. Using Xterra MS column, the mobile phases consisted of methanol:water:formic acid (25:75:0.5) at a flow rate of 0.25 ml·min-1. Beagle dogs received the intravenous dosage of PMEA-Na at 1.0, 3.0 and 6.0 mg·kg-1. Pharmacokinetic parameters were obtained from concentration-time curves by non-linear least-squares regression using the program DAS. RESULTS: The linear calibration curve was obtained in the concentration range of 0.02 to 20 mg·L-1 (r=0.999), and the limit of quantitition was 20 μg·L-1. The within-day and internal-day precisions (RSD) were less than 6.5% and 10.8%, respectively. The accuracy was 97.1%～107.3%. After a single dose studies in dogs the AUC were 2.3±0.5, 8.2±1.3 and 18.5±1.3 mg·L-1·h; the t1/2 were 3.9±1.8, 8.4±1.5 and 8.9±0.6 h; the CL were 0.44±0.09, 0.35±0.05 and 0.31±0.03 ml·h-1·kg-1 at the dose level of 1.0, 3.0 and 6.0 mg·kg-1 respectively. CONCLUSION: The analytical method is sensitive and specific for investigation the pharmacokintics of PMEA-Na in beagle dogs.

BACKGROUND: Active bone morphogenetic protein (BMP-2) reduction can cause severe symptoms like osteoporosis. In addition, there is estrogen receptor (ER) in osteoblast, osteoclast and bone marrow stromal cells, indicating a direct effect of estrogen on bone tissues.OBJECTIVE: To investigate the effect of Chinese medicine of Jiangzhi Zhuanggu on BMP-2 and ER expression in rats with prey.hyperlipidemia induced osteoporosis.METHODS: A total of 27 adult SD rats, of either gender, weighing 180-230 g, were randomly divided into three groups. In the normal control group, rats were intragastrically infused with 5 mL/kg normal saline every morning and afternoon. In the model group, the rats were infused with 5 mL/kg high-fat diet in the morning and 5 mL/kg normal saline in the afternoon. In the Chinese medicine group, 5 mL/kg high-fat diet was infused in the morning and 5 mL/kg Chinese medicine of Jiangzhi Zhuanggu water extract in the afternoon. After 8 weeks, expression levels of BMP-2 and ER in bone tissue was detected with immunohistological methods, and ER mRNA level of bone tissue in rats was detected by in situ hybridization.RESULTS AND CONCLUSION: Compared with the model group, the BMP-2 and ER expression in the bone tissue was significantly increased (P ＜ 0.01), and ER mRNA level increased following Chinese medicine treatment. Results show that Chinese medicine of Jiangzhi Zhuanggu could increase BMP-2 and ER expression in the osteoporosis bone tissue, and improve osteoporosis effectively.

AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.

Objective To study osteoporosis associated with hyperlipidemia;To research high cholesterol se-rum and Jiangzhizhuanggu-containing serum intervention rat bone marrow stromal cells BMP-2 expression changes. Methods Adult rat femur by density gradient centrifugation separation of bone marrow stromal cells, cultured cells to the third generation, and was identified by flow cytometry. The third generation cells, were randomly divided into three groups. Normal serum control group: two percent blank senun (volume ratio); High cholesterol serum injury group: fi-nal concentration of 4 mmol/L, high cholesterol serum; medicine + pretreatment serum high cholesterol serum injury, group :2% senun Chinese medicine(volume ratio) after pretreatment 2 hours + final concentration of 4 mmol/L, high cholesterol serum. Total RNA was isolated from cells recovered. BMP-2 mRNA expression was detected. And analyzed statistically. Results (1) Blank serum compared with the control group,high cholesterol serum cultured bone mar-row stromal cells proliferation was inhibited. BMP-2 mRNA expression significantly decreased(P < 0.01) . (2) Jiang-zhizhuanggu solution containing serum protection two hours later to join a high cholesterol serum group, the cell growth conditions improved significantly, BMP-2 mRNA expression significantly increased than pure high cholesterol serum injured group(P <0.01) . Conclusion Solution's Jiangzhizhuanggu containing serum can enhance high cholesterol serum environment bone manow stromal cells BMP-2 mRNA expression, significantly improved bone marrow stromal cell growth state is conducive to the treatment of osteoporosis.

Objective To investigate the effect of portal vein ligation combined with in situ splitting on liver regeneration in rats .Methods Seventy-five healthy male Sprague-Dawley rats were selected and randomly assigned into sham operation group ( S) , portal vein ligation group ( PVL) and portal vein ligation combined with in situ splitting group ( ALPPS) .On 1 d, 3 d, 7 d, 10 d, 14 d after operation , the hepatic regeneration rate ( HRR) of right median lobe was calculated , the serum alanine aminotransferase ( ALT) , aspartate aminotransferase (AST), IL-6, HGF, VEGF were detected.mRNA of IL-6, HGF, TNF-α, TGF-βwas assayed by real-time PCR, and the hepatic proliferating cell nuclear antigen ( PCNA) labeling index was evaluated by immunohistochemistry .Results Comparing with PVL group , the HRR of the right median lobe obviously increased on day 3, 7, 10 and 14 in ALPPS group (P<0.05), and ALT and AST level were increased on 1 d (P<0.05).On day 1 and 3, the content of serum IL-6, HGF and VEGF were all in-creased in ALPPS group [(70.7 ±14.6) pg/ml vs.(134.2 ±31.4) pg/ml; (0.70 ±0.04) ng/ml vs. (0.74 ±0.02) ng/ml;(82.1 ±12.6) pg/ml vs.(103.5 ±14.7) pg/ml], respectively (P<0.05).The mRNA expression of IL-6, HGF, TNF-α, TGF-βand the PCNA labeling index were also increased in ALPPS group in comparison with those in PVL group on day 1 and 3 (P<0.05).All the indexes in the two groups were all higher than those in the group S ( P<0 .05 ) .Conclusions Portal vein ligation combined with in situ splitting could significantly enhance liver regeneration .The possible mechanisms were related to the inflammation reaction and stress response caused by in situ splitting and up-regulation of cytokines in the regenerating lobe after portal vein ligation combined with in situ splitting , especially IL-6, HGF and TNF-α.

Objective To investigate the protection of Liuwei Dihuang Jiawei Capsula(LDJ Capsula,Rehmanniae Capsula of Six Ingredients) on diabetic nephropathy(DN) rats′ kidney and effect on renal protein kinase C(PKC) activity and connective tissue growth factor(CTGF) of DN rats.Methods The DN rat models were induced by ip injection of streptozotocin(STZ).The rats were randomly divided into five groups: control group; DN model group;Lotensin group;LDJ Capsula group;and Lotensis and LDJ Capsula combination group.Drug intervention term was 12 weeks.Renal ultrastructure was observed by transmission electron microscope and Masson staining.Relative kidney weight,blood glucose level,serum and urine creatinine content,creatinine clearance,excretion rate of the 24 hour urine protein,renal PKC activity,and CTGF expression in renal cortex were measured by immunohistochemistry.Results Deposition of collagen in renal of DN rats was conspicuous.Relative kidney weight,blood glucose level,serum and urine creatinine content,creatinine clearance,excretion rate of 24 h urine protein,renal PKC activity and CTGF expression of DN rats increased obviously.All Lotensin,LDJ Capsula,and the combination of these two drugs could decrease renal PKC activity and CTGF expression and ameliorate proteinuria and renal function of DN rats.At the same time they all could abate the deposition of collagen in renal of DN rats.Combination of these two drugs could decrease renal PKC activity and CTGF expression more ob-viously and at the same time had more notable protective effect on kidney of DN rats.Conclusion All Lotensin,LDJ Capsula,and the combination of these two drugs could protect kidney of DN rats.The combination of these two drugs has more obviously protective effect than using Lotensin only.

OBJECTIVE To explore the clinical efficacy of adenoidectomy in treating pediatric secretory otitis media,sinusitis and snoring. METHODS A retrospective study was conducted to analyze the clinical features,therapeutic methods and prognosis of 68 children in hospital who underwent adenoidectomy to treat secretory otitis media,sinusitis or snoring. RESULTS The conditions of the 68 children were marked improved after the removal of hypertrophic adenoids. The total clinical effectiveness rate was 100 % and the cure rate was 86.8 %. CONCLUSION Hypertrophic adenoids is a fundamental cause of pediatric secretory otitis media,sinusitis and snoring. The removal of hypertrophic adenoids is a safe and effective method for treating pediatric secretory otitis media,sinusitis and snoring.