Objective To determine whether or not the bacterial DNA which was detected by PCR comes from viable bacteria.Methods Observe the effection of middle ear effusion(MEE) on DNA viscosity and enzymatic digestion of DNA.Results The middle ear effusion and DNA are stable and DNase 1 rapidly digests DNA,The effusion does not seem to degrade DNA.The middle ear effusion signifcantly inhibits DNase 1.Conclusions Middle ear effusion provides an inhibition of the enzymatic digestion of purified DNA.Thus any DNA found in effusion by PCR techniques could well be fossilized remains and chronic otitis media with effusion may not be the bacterial infection that recent studies have suggested.

ObjectiveTo investigate the influence of cobalt chloride ( CoCl2 )-mimetic hypoxia on theproliferation,apoptosis and migration of human pancreatic cancer cell fine PANC1.MethodsPANC1 cells were treated with 0(control),100,200,400,800 μmol/L CoCl2 respectively for 24 h.Real-time RT-PCR and Western blot were used to determine hypoxia induced factor ( HIF)-1o mRNA and protein expression respectively,and cell counting kit-8(CCK-8) assays,flow cytometry and cell scratch test were used to examine the proliferation,apoptosis and migration of PANC1 cells,respectively.ResultsIn the control group and 100,200,400 and 800 μmol/L CoCl-2 groups,the expressions of HIF-1t mRNA were 1,1.08 ±0.12,1.12 ± 0.09,1.04±0.11,0.66 ±0.07,and the expressions of VEGF mRNA were 1,2.69±0.35,4.81 ±0.54,2.19 ± 0.21,0.79 ± 0.08,while the expressions of HIF-1 α protein were 0.23 ± 0.03,0.36 ± 0.04,1.15 ± 0.11,1.08 ± 0.09,0.44 ± 0.04; and the expressions of VEGF protein were 0.14 ± 0.02,0.12 ± 0.01,0.95 ±0.09,0.87 ±0.09,0.55 ±0.06; and cell viability rates were 100％,(98.43 ±2.88)％,(76.15 ± 0.70)％,(53.87 ±0.77)％,(35.23 ±0.67)％ ; while cell apoptotic rates were (5.2 ±1.12)％,(5.74 ± 1.07)％,(6.82 ± 1.85)％,(12.09 ±3.53)％,(31.88 ±6.95)％ ; the cell migration distance of PANC1 cells were (43.24 ±3.67)％,(59.46 ±5.39)％,(80.56 ±8.05)％,(63.89 ±5.96)％,(9.09 ± 1.59 ) ％.Compared with those of control group,the expressions of VEGF mRNA,VEGF and HIF-1 α protein,cell migration distance showed a two-way variation ( ascending first and descending later) (P ＜0.05 ),and the expression of HIF-1α mRNA and cell proliferation rate was decreased in a dose-dependent manner,while the cell apoptosis was increased in a dose-dependent manner.Conclusions CoCl2 significantly inhibits the proliferation and promotes apoptosis of PANC1 cells at certain level.CoCl2 has a two-way effect on the migration of PANC1 cells,and it may be related to the direct injury of high concentration of CoCl2 on cells.

Objective To investigate the expressions of RGC-32 and E-cadherin in pancreatic cancer and analyze their clinicopathological significance and the correlation with each other.Methods SP immunohistochemistry was used to detect the expressions of RGC-32 and E-cadherin in 42 cases of pancreatic cancer tissues,12 cases of chronic pancreatitis tissues and 8 cases of normal pancreatic tissues.Results The positive staining for RGC-32 was predominantly observed in the cytoplasm of pancreatic acinar cells.The positive staining for E-cadherin was mainly observed in the cytomembrane of normal pancreatic and chronic pancreatitis acinar cells,but aberrant expression ( cytoplasm expression and ( or ) weaker expression) could be found in pancreatic cancer cells.The positive expression rate of RGC-32 and aberrant expression rate of E-cadherin were 78.6％ (33/42) and 54.8％ (23/42),respectively,in pancreatic cancer tissues,which were significantly higher than those in normal pancreatic tissues [37.5％ (3/8) and 0] and chronic pancreatitis [41.7％ (5/12)and 8.3％ (1/12) with statisticai significance,P ＜0.05].The expression of RG C-32 in pancreatic cancer was associated with lymph node metastasis and TNM staging (P =0.016,0.025,respectively),but not with age,gender and differentiation degree ( P =0.831,1.000,0.629,respectively).The aberrant expression of E-cadherin was associated with differentiation degree,lymph node metastasis and TNM staging ( P =0.024,0.004,0.004,respectively),but not with age and gender ( P =0.970,1.000,respectively).A significantly positive correlation was found between positive expression rate of RGC-32 and aberrant expression rate of E-cadherin (r =0.458,P ＜0.01 ).Conclusions Both positive expression rate of RGC-32 and aberrant expression rate of E-cadherin are up-regulated significantly in pancreatic cancer tissues and RGC-32 may be involved in the invasion and metastasis of pancreatic cancer by regulating epithelial mesenchymal transition.

OBJECTIVEAccidents are an important cause of childhood injury. It is hypothesized that safety education programs can reduce accidents in primary school-aged children. This study aimed to determine whether child and parent safety education programs can decrease the incidence of accidental injury in children when compared with controls.

METHODSThe study population (aged 7-13 years) were recruited from four local primary schools, and randomly assigned into an Intervention or a Control group. The Intervention group received child and parent safety education and was taught injury prevention strategies. The Control group received no injury prevention education or intervention. The incidence of accidental injury was compared between the two groups.

RESULTSIn the first year after intervention the incidence of accidental injury was 262 cases in the Intervention group (8.26%) and 234 cases (8.67%) in the Control group (P > 0.05). In the second year after intervention, however, the incidence of accidental injury was significantly less in the Intervention group (211 cases, 6.54%) compared with the Control group (229 cases, 8.63%) (P < 0.01).

CONCLUSIONSInjury prevention strategies and child and parent safety education can reduce risks of accidental injury in children.

Accident Prevention ; methods ; Adolescent ; Child ; Female ; Humans ; Male ; Parents ; Safety ; Wounds and Injuries ; prevention & control

This paper is to evaluate the biocompatibility and cytotoxicity of a new Ni-free Zr-based bulk metallic glass (BMG), Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, by comparing it with conventional Ti6Al4V alloy. According to ISO 10993-5: 1999 and GB/T 16886.5-1997 standards, Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3, pure Zr and Ti6Al4V materials were extracted with surface area of sample/volume of medium ratio being 1 cm2/mL and 0.5 cm2/mL, respectively. The viabilities of MG-63 cells (Human osteosarcoma cell line) cultured in the BMG medium extracts for 1, 3 and 5 days were determined by CCK-8 assay. The cellular morphology of MG-63 cells cultured on the surface of samples for 3 days was tested through laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM). The relative growth rate (RGR) of MG-63 cells cultured in Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and pure Zr were both more than 85%, indicating that the cytotoxicity of BMG was relatively low and met the national biomedical material eligibility standard. There was insignificant difference in the morphology of MG-63 cells cultured in the BMG medium extracts and the control group through LSCM and SEM, which showed the BMG had excellent biological compatibility. The Zr-based bulk metallic glass Zr60.14 Cu22.31 Fe4.85 Al9.7 Ag3 and the conventional Ti6Al4V alloy both had no obvious cytotoxicity to MG-63 cells. These results provided evidence that the new Zr-based bulk metallic glass could be potential replacement material for the orthopedic surgical implant.
Alloys ; Biocompatible Materials ; Cell Line ; Glass ; Humans ; Microscopy, Electron, Scanning ; Nickel ; Titanium ; Zirconium

Objective To investigate the effect of genistein on Notch-1, SHH and HHIP gene expression and on the cell cycle and proliferation of of BxPC3 cells. Methods Human pancreatic cancer cell line BxPC3 was cultured. The BxPC3 cells were treated with genistein and then the total RNA and protein were extracted. RT-PCR was used to detect the expression of Notch-1 mRNA, SHH mRNA and HHIP mRNA. Noteh-1 and SHH protein was determined by western blotting. MTT assay was used to detect proliferation of BxPC3 cells. The cell cycle of BxPC3 cells was measured by Propidium iodide (PI) and flow cytometry. Results The inhibiting rate was 67.17%±2.32% when BxPC3 cell lines were treated by 20μg/ml genistein for 48 hours. Notch-1 mRNA was down-regulated from 2.454±0.068 to 1.304±O.169 ; SHH mRNA was down-regulated from 0.959±0.023 to O.472±0.077 ; HHIP mRNA was up-regulated from 0.625±O.158 to 1.761±0.121. Notch-1 protein expression was down-regulated from 1.361±0.109 to 0.760±0.114; SHH protein expression was down-regulated from 0.265±0.018 to 0.129±0.013. (52.77±9.47)% cells were hindered in G2/M stage. Conclusions Genistein could down-regulate Notch-1 expression and inactivate Hedgehog signaling pathway and inhibit the proliferation of pancreatic cancer cells.

To evaluate the status of diabetes-related knowledge and training requirement among general physicians,a questionnaire survey was conducted in 676 general physicians from 32 primary medical institutions and 5 secondary hospitals of Shanghai Pudong New Area.48.2% (261/541) of the surveyees had bachelor degree and most of them worked in secondary hospitals or primary medical institutions in affluent areas.The awareness rate of diabetic knowledge among the surveyeos was 51.6%,and there were significant differences in the awareness rate of diabetes-related knowledge between different levels of hospitals and physicians with different academic background.Only 26.6% of physicians received diabetes-related continuing education or training,there was a popular request for professional training from the surveyed physicians.

Objective To observe the expression of survivin and Aurora B in human pancreatic cancer BXPC3 cells after the treatment of sulindac and to explore the potential mechanism. Methods MTr assay was used to determine the effect of sulindac on the proliferation of the BXPC3 cells. RT-PCR was used to detect the expression of mRNA level of survivin and Aurora B, western blot was used to detect protein expression of survivin and Aurora B Thr-232. Cell cycle and apoptosis were detected by flow eytometry (FCM). Results The BXPC3 cells were inhibited by sulindac in a dose and time-dependent manner; the expression of mRNA of survivin and Aurora B were both significantly decreased from 1.5644 and 0.6554 to 0. 4372 and 0.1132 (P< 0.01), the expression of survivin protein and the phosphorylation of Aurora B Thr-232 were also decreased from 1.2735 and 0.4680 to 0.2126 and 0.2546 (P<0.01); the proportion of cells in the G0/G1 phase was increased from (56.65±1.93)% to (70.58±3.21)% (P<0.01). Conclusions Sulindac had inhibitory effects on the growth of BXPC3 cells, the possible mechanism was via decreasing the expression of survivin which depressed the activity of Aurora B, then the CPC was influenced. The most of the cells were blocked in the G0/G1 phase, and the cells' mitosis was inhibited.

Objective To observe prospectively the role of endoscopic diagnosis and treatment of biliary leakages in patients with liver transplantation, and the incidence of bile duct stricture after healing of the leakage. Methods Six eases of T-tube leakage and seven cases of anastomosis leakage complicating liver transplantation were enrolled in this prospective study. Six patients treated by endoscopic plastic stent placement , 2 by naso-biliary catheter drainage, 2 by papillosphincterotomy and 3 by naso-biliary catheter drainage combined with plastic stent placement. Some patients received growth hormone treatment. Results The bile leak resolution time was between 10-35 days in 10 patients with complete document. The median time of leak resolution was 15. 3 days. Four cases of anastomosis stricture, three cases of common hepatic duct and one ease of multiple bile duct stenosis were observed by followed-up nasobiliary catheter cholangiography or ER-CP. Conclusion Endoscopic nasobiliary catheter or plastic stent placement is a safe and effective treatment for bile duct stricture occurred after bile leak resolution in most of liver transplantation patients. Naso-biliary catheter combined with plastic stent placement maybe the best choice for treating bile leak, because, theoretically, it may prevent serious condition happened at accidental nasobiliary catheter dislocation, and it may have prophylactic effect on upcoming bile duct stricture and should be further confirmed.

Objective To observe the influence ofgypenosides on hydrogen sulfide in liver tissue and plasma of rat with type 2 diabetes mellitus and nonalcoholic fatty liver disease.Methods 58 SPF male SD rats,with body mass 220～250 g,were randomly divided into a blank control group (group N,n=7),and a NAFLD and T2DM model group (Group M,n=51).Group N was fed with ordinary diet in the first four weeks,group M was fed with diets of high fat and sugar,injected with 40 mg/kg STZ overnight,and the same diets for the next four weeks.The rat model with T2DM and NAFLD was build.NAFLD and T2DM model group were divided into three groups:a high dose GPS group (JH,n=9) injected with 1 g/kg · d-1 GPS,a low dose GPS group (JL,n=9) injected with 0.5 g/kg · d-1 GPS,and a model group (M,n=9) injected with the same volume of water,and high fat diet at the same time.The treatment period was six weeks,and the experiment period was fourteen weeks.TG,TC,BS,and H2S in the plasma of rat were tested,and H2S in the liver tissue of the rat was tested.Results ①The changes of H2S in plasma:group JH [(4.30±0.43) μmol/L] and JL [(3.83 ±0.47) μmol/L] was lower than group M [(2.67 ± 0.41) μmol/L],there was a significant difference.②The changes of H2S in the liver tissue:group JH [(333.52±37.94) pmol/min/mg/protein] and JL [(275.81 ±36.07)pmol/min/mg/protein] was lower than group M [(237.8± 33.05) pmol/min/mg/protein],there was a significant difference.③BS levels:group JH(10.86±3.46)mmol/L,group JL (14.78±3.39)mmol/L,group M(18.84±4.24) mmol/L,group JH and JL was lower than group M,there was a significant difference (P＜0.01).④The plasma TG level:group N (0.96±0.09) mmol/L,group JH (2.82± 0.66) mmol/L,group JL (1.83± 0.56) mmol/L,group M (3.97 ± 0.64) mmol/L.group JH and JL was lower than group M,there was a significant difference (P＜0.01).Conclusion Gypenoside can reduce the blood sugar,triglycerides,and total cholesterol in rat with with type 2 diabetes mellitus and nonalcohol fatty liver disease.H2S concentrations in plasma and liver tissue of the rats with T2DM and NAFLD were increased by GPS,showing dose dependence.Gypenosides can also improve metabolism of blood glucose and lipid in rats with T2DM and NAFLD.