The voltage-gated potassium channels Kvl.5 is widely expressed in the plasma membranes of numerous tumor cells and it can contribute to a variety of cellular functions such as proliferation,ap optosis,and cell cycle.Several types of antineoplastic drugs can change the expression of Kv1.5 and then affect the biological processes.Kv1.5 is identified as a novel target for therapy in human cancer.The researches of Kv1.5 will contribute to gain a further understanding of the molecular mechanism of tumor and provide therapeutic opportunity for the prevention and treatment of cancer.

Objective To detect the expression of HERG (human ether-à-go-go-related gene ) potassium channel in human osteosarcoma,and explore the effects of silencing HERG by small interfering RNA (siRNA)on the proliferation and apoptosis of osteosarcoma cells and the mechanisms responsible for HERG regulation.Methods The expressions of HERG in osteosarcoma MG-63 cells and tissues were detected by reverse transcription polymerase chain reaction (RT-PCR),Western blotting and immunohistochemistry.Next, osteosarcoma cells were divided into three groups:HERG-siRNA group,control-siRNA group and blank group. CCK-8,colony formation,flow cytometry and Tunel assay were used to measure the proliferation and apoptosis of the osteosarcoma cells.Finally,Western blotting analysis was performed to detect the expression of nuclear factor-κB (NF-κB)pathway in osteosarcoma cells treated with HERG siRNA.Results Osteosarcoma cells and tissues were found to highly express HERG.Inhibition of HERG in the osteosarcoma cells significantly inhibited the cell proliferation and induced cell apoptosis.Compared to control-siRNA group or blank group,HERG-siRNA could inhibit the proliferation of MG-63 cells significantly [HERG-siRNA group:(75.34 ± 4.45)%;compared to control-siRNA group:(100.60 ±5.31)%;t =3.64,P =0.007;compared to blank group:(100.00 ±5.66)%;t =3.43,P =0.009].The similar results were obtained from colony formation assay (HERG-siRNA group:134.30 ±11.82;compared to control-siRNA group:225.30 ±11.56;t =5.51, P =0.002;compared to blank group:232.80 ±12.21;t =5.80,P =0.001).HERG-siRNA transfected MG-63 cells demonstrated a significant increase of apoptotic rate compared to control-siRNA transfected cells or untreated cells [HERG-siRNA group:(28.10 ±2.21 )%;compared to control-siRNA group:(9.36 ± 2.42)%;t =5.72,P =0.005;compared to blank group:(10.92 ±2.51)%;t =5.14,P =0.007].This resultwas further confirmed by Tunel assay.The cells transfected with HERG-siRNA (31.57 ±2.08)% dem-onstrated extensive apoptosis,compared with the control-siRNA group [(10.35 ±1.82)%;t =7.69,P =0.002)]or blank group [(7.96 ±0.88)%;t =10.48,P =0.001].Silencing HERG gene down-regulated the cIAP-1,XIAP,Bcl-2,Survivin,P-IκBαand NF-κB p65 expression,compared to the control groups. Conclusion HERG is highly expressed in osteosarcoma.HERG silencing can suppress osteosarcoma progres-sion through NF-κB pathway and suggest that HERG may be a novel molecular target for osteosarcoma therapy and diagnosis.

Objective To research a new type of gastrointestinal anastomat -- anastomosis device with three rows of staples (Pa-tent No .2012200583213) ,and clarify its associated functions through esophagogastric anastomosis operations on pigs ,so that we could provide experiences for its clinical application in future .Methods Compared with domestic anastomat with two rows of sta-ples ,we designed and manufactured a new type of anastomat with three rows of staples and researched its function .Choosing 12 do-mestic pigs ,about 60 kg ,gastroesophageal anastomosis was taken twice with each case by anastomat with three or two rows of sta -ples randomly in sequence .According to the different types of anastomats ,cases were divided into two groups :group A ,used anas-tomat with three rows of staples ,including 12 cases of anastomosis ;group B ,used anastomat with two rows of staples ,including an-other 12 cases of anastomosis .Results Compared with group B ,cases of group A have less bleeding sites (t = 7 .00 ,P < 0 .01) . Without reinforcement and with 0 .5 kg of tension ,fewer of outermost staples exposed(t= 6 .17 ,P< 0 .01) .And the shape of used staples of group A is double circles ,which has bigger mechanical strength than that of group B (t= 6 .57 ,P < 0 .01) .Conclusion The function of anastomat with three rows of staples surpasses that of traditional anastomat with two rows of staples in pig esopha -gogastrostomy surgery .

Objective To establish and assess the utility of 99Tcm-sulfur colloid (SC) salivary imaging in the routine evaluation of pulmonary aspiration in adult patients with respiratory tract diseases.Methods Eight patients (7 men,1 woman; age range 68 to 80 years,mean age (76 ± 4) years) with respiratory tract disease and history of aspiration by clinical assessment were evaluated prospectively by 99Tcm-SC salivary imaging from April to July 2012.A dose of 74.0 MBq 99Tcm-SC was added to 20 ml saline,mixed well,and administered orally to patients.Dynamic imaging was acquired with posterior projection for 30 min at a rate of 30 s per frame.Two experienced physicians assessed all examination results and reached consensus for final diagnosis.Radioactivity detected at either the bronchi or within the lung fields was reported as positive for aspiration.This study was approved by the institutional review board of Hospital Ethical Committee,and the written informed consent was obtained from patients or their guardians.Results All patients were positive for aspiration by 99Tcm-SC salivary imaging (8/8).Aspiration into bilateral main bronchus was seen in 2 cases,right main bronchus and branch in 4 cases,and left main bronchus and branch in 2 cases.Aspirated tracer could be visualized as early as 3 min,latest at 24 min,and the median was 19 min.Conclusion 99Tcm-SC salivary imaging is useful for the detection of aspiration in adult patients with respiratory tract diseases.

Objective To study the effect of endothelial progenitor cells (EPCs) transplantation on CCl4 induced hepatic cirrhosis in rats. Methods Eight male SD rats were used as normal control. Thirty rats were induced liver cirrhosis by feeding with 25% CCl4/olive oil for 12 weeks, and then were subdivided into cirrhosis group (n = 10), EPCs transplanted group (n = 10) and saline control group (n = 10). EPCs were transplanted into the portal vein for 4 weeks in EPCs transplanted group. Rats in EPCs nontransplanted group were sacrificed at the beginning of the 12th week. Rats in EPCs transplanted group and saline control group were killed at the beginning of 16th week. Serum biochemical parameters were examined. The degree of liver cirrhosis was evaluated by Masson staining and by detecting the expression of α-SMA, Collagen Ⅲ and Ki67. Results The volumes of liver in cirrhosis group were twice as much as that in normal rats. 12 weeks after CCl4 administration, compared with saline control group, in EPCs transplanted group, hepatic activity index (HAI) ( F = 75. 062, P < 0. 01 ), the levels of ALT( F = 29. 942, P<0.05), AST(F=16.618,P<0.05) and TBIL(F=9.911 ,P<0.05) in serum decreased, the level of Alb ( F = 4. 944, P < 0. 05 ) and Ki67 ( F = 45. 966, P < 0. 01 ) was increased, the expression of α-SM A ( F = 7.86,P<0.05) and collagen Ⅲ (F = 135.787,P <0.01) decreased (P <0.05). Compared with untransplanted group, in EPCs transplanted group, the levels of ALT, AST and TBIL in serum were lower; In saline control group, the levels of ALT, AST and TBIL in serum were higher, the level of Alb and Ki67was lower, the expression of α-SMA and collagen Ⅲ were higher( P < 0. 05 ). Compared with normal rats, in saline control group, the levels of INR were higher (P < 0. 05 ). Conclusion EPCs transplantation improves hepatocye regeneration and ameliorates established hepatic cirrhosis.

Objective To compare the disinfection efficacy of different disinfectants on dental unit waterlines (DUWLs). Methods 18 sets of DUWLs were randomly divided into 4 groups,and disinfected or treated with hydrogen peroxide (H2 O2 )disinfectant,sodium hypochlorite (NaClO)disinfectant,hydrogen peroxide silver ion disinfectant(Sanosil),and distilled water (DW)respectively.Water specimens from triple syringes and high-speed handpieces were taken,bacterial count before and after disinfection were compared.Results Before disinfection,no significant differences in bacterial counts were found among four groups (all P >0.05),bacterial counts of DUWLs of all groups severely exceeded the standard(all>3 000 CFU/mL).After disinfection,except DW group,bacterial counts of DUWLs of the other groups declined dramat-ically (all <100 CFU/mL),bacterial count after disinfection were all obviously lower than before disinfection (all P <0.001 ).One week after disinfection,bacterial counts among three disinfectant groups in different time periods were statisti-cally different (triple syringes:Day1—Day5,all P <0.05;high-speed handpieces:Day2,Day3 and Day5,all P <0.05). Day3 after disinfection of triple syringes by H2 O2 and NaClO,Day4 after disinfection of high-speed handpieces by H2 O2 and NaClO,and Day5 of triple syringes and high-speed handpieces by Sanosil all exceeded the standard of Center for Disease Control and Prevention of America.One week after disinfection,bacterial counts of three disinfection groups all exceeded or approximated to that before disinfection.Conclusion Three types of disinfectants can all effectively reduce bacterial load in DUWLs.Compared with other disinfectants,Sanosil has advantage of inhibiting bacterial growth after disinfection.

Objective Routine perioperative intravenous antimicrobial agents,was administered as surgical prophylaxis.However,whether balanced ultrafiltration during extracorporeal circulation can remove antimicrobial agent remains unclear.The concentrations of antimicrobial agent in plasma and ultrafiltrate samples were measured in this pseudo-extracorporeal circulation model.Methods Extracorporeal circulation consisted of cardiotomy reservoir (Ningbo Fly Medical Healthcare CO.,LTD.Ningbo,China),D902 Lilliput 2 membrane oxygenator (Sorin Group Asia Pte Ltd,Beijing,China) and Capiox (R) AF02 pediatric arterial line filter (Terumo Corporation,Beijing,China).HEMOCONCENTRATOR BC 20 plus (MAQUET Cardiopulmonary AG,Hirrlingen,Germany) was placed between arterial purge line and oxygenator venous reservoir.Fresh donor human whole blood was added into the circuit and mixed with Ringer's solution to obtain a final hematocrit of 24％-28 ％.After 30 minutes of extracorporeal circulation,zero-balanced ultrafiltration was initiated and arterial line pressure was maintained at approximately 100 mm Hg(1 mm Hg =0.133 kPa) with Hoffman clamp.The rate of ultrafiltration (12 ml/min) was controlled by ultrafiltrate outlet pressure.Identical volume of plasmaslyte A was dripped into the circuit to maintain stable hematocrit during 45 minutes of experiment.Plasma and ultrafiltrate samples were drawn every 5 minutes and concentrations of antimicrobial agent (including Cefmetasole and cefotiam) were measured with high performance liquid chromatography.Results All these two antimicrobial agents were detected in ultrafiltrate,demonstrating hemoconcentration may remove antimicrobial agent.The concentration of plasma antimicrobial agent decreased lineally with the increase of ultrafiltrate volume.At end of balanced ultrafiltration,the concentration of plasma cefotiam was (104.96 ± 44.36) μg/ml,which is about (44.38 ± 7.42) ％ of the initial concentration (238.95 ± 101.12) μg/ml; the concentration of plasma cefmetazole decreased linearly to (25.76 ± 14.78) μg/ml,which is about (49.69 ± 10.49) ％ of the initial concentration (51.49 ± 28.03) μg/ml.The total amount of cefotiam in ultrafiltrate is (27.16 ± 12.17)％ of the total dose administered,whereas cefmetasole in ultrafiltrate is (7.74 ±4.17)％.Conclusion Balanced ultrafiltration may remove antimicrobial agent from serum and has significant influence on plasma concentration of antimicrobial agent.The strategy of surgical prophylaxis should consider this unique technique during extracorporeal circulation.

Objective To detect the influence of miRNA-34a (miR-34a) on the proliferation of osteosarcoma and the mechanisms responsible for miR-34a regulation.Methods The osteoblastic cell line MG-63 and Saos-2,human osteoblastic cell line hFOB 1.19,10 osteosarcoma tissues and 10 normal bone tissues were selected.The expression of miRNA-34a in osteosarcoma cells and tissues was detected by quantitative real-time polymerase chain reaction (qPCR).Next,a eukaryotic expression vector named pcDNA/miR-34a was constructed.Then,osteosarcoma cells were transfected with this eukaryotic expression vector and the effects of miR-34a overexpression on the proliferation and growth of osteosarcoma were measured using CCK-8,colony formation and xenograft model of nude mice.Finally,Western blot analysis was used to detect the expression of ether-à-go-go 1 (Eag1) gene in osteosarcoma cells after transfected with pcDNA/miR-34a or a miR-34a inhibitor miR-34a-2'-O-Methyl antisense oligoribonucleotide (miR-34a-2'-O-Me).Results Compared with normal bone tissues and osteoblastic cell line,miR-34a was down-regulated in osteosarcoma cell lines and tissues.Compared with the blank group and the control group,the cell survival rates of miR-34a group of the two cell lines were significantly lower [MG-63 72 h:blank group (40.05±4.82) ％,control group (36.88± 4.66) ％,miRNA-34a group (26.24±6.22) ％;MG-63 96 h:blank group (83.55±5.95) ％,control group (80.13± 4.48) ％,miRNA-34a group (30.21±7.26) ％;Saos-2 72 h:blank group (46.45±8.15) ％,control group (43.33± 6.89) ％,miRNA-34a group (26.81±3.17) ％;Saos-2 96 h:blank group (84.79±4.10) ％,control group (80.14± 3.11) ％,miRNA-34a group (31.77±5.17) ％].The similar results were obtained from colony formation assay (MG-63:blank group 83.40±3.29,control group 80.00±3.06,miR-34a group 24.40±2.71;Saos-2:blank group 85.00±3.32,control group 80.60±3.29,miR-34a group 30.40±4.94).The tumor volumes of osteosarcoma xenograft in the miR-34a group was significantly smaller than that in the blank group and control group after 21 days treatment (all P ＜ 0.001).Overexpression of miR-34a could decrease Eag1 expression in osteosarcoma cell lines while inhibition of miR-34a induced the of expression Eag1 (P ＜ 0.001).Conclusion MiR-34a plays a tumor suppressor role in osteosarcoma and could suppress the proliferation and growth of osteosarcoma through the regulation of Eag1.Moreover,it may be a novel target for osteosarcoma therapy.

Objective To investigate the clinical efficacy of our self-designed adjustable weight-bearing brace for AO type B tibial shaft fractures managed by interlocking intramedullary nail.Methods A total of 68 consecutive patients with AO type 42-B tibial shaft fracture who had been managed from April 2013 to March 2015 hy interlocking intramedul]ary nail were recruited into our study.They were randomized into 2 equal groups (n =34).Group A received conventional therapy after operation while group B received auxiliary mauagement with our self-designed adjustable weight-bearing brace after conventional postoperative therapy for one week.The 2 groups were compared at postoperative 1,3 and 6 months and at the final follow-up in terms of visual analogue scale (VAS),weight-bearing status of the affected limb,time for fracture union,Radiographic Union Score for Tibial Fractures (RUST) and Johner-Wruhs scale.Results Of this series,62 cases were followed up for 12 to 18 months (average,14.7 months),5 ones were lost to the follow-up and one withdrew.The mean VAS scores at 3-month and 6-month follow-ups for group B were 2.5 ± 0.8 and 0.9 ± 0.6 respectively,significantly lower than those for group A (3.0 ± 0.9 and 1.4 ± 0.8 respectively) (P ＜ 0.05).In group A at 1-month,3-month and 6-month follow-ups,the weight-bearing status was 44.1％ ± 17.5％,72.0％ ±17.4％ and 86.4％ ±12.5％ while the mean RUST scores were 5.4±1.4,8.7±1.1 and 10.3 ± 1.1,respectively.In group B at 1-month,3-month and 6-month follow-ups,the weight-bearing status was 53.8％ ± 11.0％,84.1％ ± 12.2％ and 94.4％ ± 10.6％ while the mean RUST scores were 6.5 ± 0.8,9.9 ± 0.9 and 11.3 ± 0.8,respectively.There were significant differences between the 2 groups in the above indexes (all P ＜ 0.05).Group B achieved clinical fracture union after an average of 3.3 ±0.7 months,significantly faster than group A (3.9 ± 1.0 months) (P ＜ 0.05).According to the Johner-Wruhs scoring,group A had 19 excellent cases and 12 good ones while group B had 27 excellent ones and 4 good ones,showing a significant difference between the 2 groups (P ＜ 0.05).Conclusions Early application of our self-designed adjustable weight-bearing brace for patients with AO type B tibial shaft fracture managed by interlocking intramedullary nail can reduce postoperative pain,accelerate callus growth,shorten bony healing time and achieve satisfactory functional recovery.

Objective:To investigate the influence of all-trans-retinoic acid (ATRA) on the proliferation and invasion of osteosarcoma 143B cells and its possible regulatory mechanism.Methods:Different concentrations of ATRA were used to treat human osteosarcoma 143B cells, and the optimal concentration and treatment time those affected cell proliferation were selected. The MTS method, Transwell migration and invasion experiments were used to detect the changes in the proliferation, migration and invasion of 143B cells after ATRA treatment. The real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression changes of miRNA-34a (miR-34a), E2F1 and Eag1 in osteosarcoma 143B cells after ATRA treatment. Then miR-34a was interfered and E2F1 was overexpressed, and the abilities of cell proliferation, invasion and migration abilities as well as the expression changes of miR-34a, E2F1 and Eag1 in 143B cells were detected.Results:The proliferation inhibition of 143B cells was most obvious when 143B cells were treated with 10 μmol/L ATRA for 72 h. The cell migration and invasion numbers when 143B cells were treated with 10 μmol/L ATRA for 72 h were lower than those in the negative control group [(73±3) cells vs. (182±5) cells, t = 21.46, P<0.01; (94±3) cells vs. (203±7) cells, t = 13.70, P<0.01]. 10 μmol/L ATRA could promote the expression of miR-34a in 143B cells and inhibit the expressions of Eag1 and E2F1 (all P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+miR-34a interference group was restored after 72 h of treatment [cell survival rate (41.0±2.2)% vs. (25.0±3.6)%, t = 108.68, P<0.01]. Compared with ATRA group, the abilities of cell migration and invasion in ATRA+miR-34a interference group were restored [(122±14) cells vs. (64±10) cells, t = 21.06, P<0.01; (103±10) cells vs. (59±8) cells, t = 24.27, P<0.01), and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Compared with ATRA group, the ability of cell proliferation in ATRA+E2F1 overexpression group was restored [cell survival rate (40.0±3.4)% vs. (24.0±3.1)%, t = 108.74, P<0.01]; the abilities of cell migration and invasion in ATRA+E2F1 overexpression group were restored [(78±12) cells vs. (29±8) cells, t = 13.52, P<0.01; (75±12) cells vs. (49±10) cells, t = 6.28, P<0.01], and the mRNA and protein expressions of Eag1 and E2F1 in cells were promoted (both P<0.01). Conclusion:ATRA inhibits the proliferation and invasion of osteosarcoma cells via regulating miR-34a-E2F1-Eag1 signaling pathway, and it may become one of the effective treatment drugs for osteosarcoma.