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.

Objective:To develop an HPLC-MS/MS method for the determination of rosuvastatin in plasma and study the relative bioavailability and bioequivalence of the capsules and tablets in Chinese healthy volunteers. Methods: A single oral dose (20 mg of the test or reference preparation) was given to 24 male healthy volunteers in a randomized crossover study. The plasma concentration of rosuvastatin was determined by HPLC-MS/MS. The pharmacokinetic parameters were calculated and the bioavailability and bioequiva-lence of the two preparations were evaluated by DAS 3. 0 software. Results:After a single dose, the pharmacokinetic parameters of ro-suvastatin capsules and tablets were as follows:Tmax was (3. 56 ± 1. 68) h and (3. 63 ± 1. 56) h, Cmax was (21. 17 ± 13. 74) ng· ml-1 and (26.33 ±23.22) ng·ml-1, t1/2 was (10.68 ±5.50) h and (9.04 ±6.00) h, AUC0-t was (219.31 ±146.09) ng·h· ml-1 and (252. 43 ± 194. 96) ng·h·ml-1 , AUC0-∞ was (225. 32 ± 146. 76) ng·h·ml-1 and (257. 24 ± 194. 61) ng·h·ml-1 , respectively. The 90% confidential interval of AUC0-t, AUC0-∞ and Cmax was 81. 1%-106% , 81. 8%-105. 4% and 77. 9%-104. 5%, respectively. The mean relative bioavailability of the test preparation(the capsules) to the reference preparation(the tablets) was (100. 7 ± 54. 1)%. Conclusion:The test and reference preparations are bioequivalent.

Objective To detect the PML-RARα fusion gene expression of APL patients with FISH and fluorescent quantitative PCR and discuss the sensitivity and specificity of two techniques. Methods The detection of the PML-RARα fusion gene expression of 75 APL patients with FISH and fluorescent quantitative PCR were carried out simultaneously. Results Eighty eight patients of primary and relapse or remission phases were examinated and total conformity rate was 96.59 %. Fourteen primary patients were detected and conformity rate was 100 %. Seventy four relapse or remission patients were detected and conformity rate was 95.95 %. Stem cell essays were detected for six times and conformity rate was 100 %. Conclusion The sensitivity of FISH and fluorescent quantitative PCR is identical for primary APL patients and FISH is more sensitive. But the sensitivity of FISH is weaker than that of fluorescent quantitative PCR for detection of residue disease.

In order to investigate the effect of nerve growth factor (NGF) on the proliferation of rabbit corneal endothelial cells and epithelial cells, the in vitro cultured rabbit corneal endothelial cells and epithelial cells were treated with different concentrations of NGF. MTT assay was used to examine the clonal growth and proliferation of the cells by determining the absorbency values at 570 nm. The results showed that NGF with three concentrations ranging from 5 U/mL to 500 U/mL enhanced the proliferation of rabbit corneal endothelial cells in a concentration-dependent manner. 50 U/mL and 500 U/mL NGF got more increase of proliferation than that of 5 U/mL NGF did. Meanwhile, 50 U/mL and 500 U/mL NGF could promote the proliferation of the rabbit corneal epithelial cells significantly in a concentration-dependent manner. However, 5 U/mL NGF did not enhance the proliferation of epithelial cells. It was suggested that exogenous NGF can stimulate the proliferation of both rabbit corneal endothelial and epithelial cells, but the extent of modulation is different.
Cell Proliferation/*drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelium, Corneal/*cytology ; Epithelium, Corneal/*cytology ; Nerve Growth Factor/*pharmacology

AIM:To analyze the killing efficiency of six kinds of contact lens solutions and solutions with arilin on free living Acanthamoeba culturedin vitroMETHODS:Six kinds of contact lens solutions were added into 96-well microtiter plates,respectively,with each care solutions used 48 holes of them.Suspension of Acanthamoeba were added into 24 of these holes.and arilin gutta and suspension of Acanthamoeba were added into the other 24 holes.After standing in room ternperature for 8 hours,the morphologic change and quantity of the remnant Acanthamoeba were observed under the jnverted microscope.The remnant Acanthamoeba were cultivanted in peptone-yeast extract-glucose (PYG)-culture medium for 5 days.Their variation of appearance,activity and reproductive activity were observed.RESULTS:In the six experimental groups using contact lens solutions only.the detection rate of Acanthamoeba of were 0%,80.3%,29.1%,41.7%,62.5% and 79.2%,respectively.After arilin was added,the detection rate of Acanthamoeba of the six groups were 0%,0%,4.2%,8.3%,16.7% and 16.7%,respectively.From group 3 to group 6,after arilin was added,the differences of the killing efficiency of contact lens solutions have statistical significance(X2=3.75,7.11,10.54 and 18.78;P＜0.05).Cultivation of the remnant Acanthamoeba showed a reduction in the activity and proliferative ability.CONCLUSION:The killing efficiency of some contact lens solutions on free-living Acanthamoeba were not satisfying.Arilin can improve the killing efficiency of contact lens solutions.

AIM: To compare the adherent ability of Acanthamoeba to three kinds of contact lens (CL) and survey the effect to remove the Acanthamoeba from the surface of contact lens after blowing with stroke-physiological saline solution (SPSS) repeatedly.METHODS: Three kinds of contact lens were chosen in this experiment, including: rigid gas permeable (RGP) contact lens, soft contact lens (SCL) and colored contact lens. They were co-cultivated with Acanthamoeba suspension for 16 hours. Then the number of Acanthamoeba adhered on CL was compared. Comparison was also made between different zones of colored CL. We also compared the number of Acanthamoeba adhered on CL in three groups after blowing with SPSS.RESULTS: The number of Acanthamoeba adhered to colored CL was more than RGP group and SCL group (P<0.05). The differences between RGP group and SCL group had no statistical significance (P>0.05). In colored CL group, the number of Acanthamoeba adhered to the colored zone was more than the Uncolored zone (P<0.01). In all the three groups, after blowing with SPSS, there was statistically significant decrease of the number of Acanthamoeba adhered to CL (P<0.01).CONCLUSION: Compared with RGP group and SCL group, the colored contact lens was more vulnerable to be adhered by Acanthamoeba. After being blown by SPSS repeatedly, the effect to eliminate Acanthamoeba has been improved greatly.

The ability to induce hypersensity on leaves of tomato and the stability of double-plasmid of an harpin-producing, nitrogen-fixing engineered strain E4 were tested. Hypelsensitivity-inducing experiment indicated that the time and density of hypersensitivity-induction of E4 was similar to those of DH5, the positive control of pCPP430. Although E4 took the same time to induce hypersensitivity as 308R, another positive control of pCPP430, it induced weaker hypersen- sitivity on tobacco leaves. On tomato leaves, there was no difference in time and density of hypersensitivity between E4 and 308R (pCPP430). Results revealed that the two plasmids, pCPP430 and pMC73A, were unstable in host bacteria, with the losing rate of 100% at the 48th generation. The emergence probability of bacteria with either pCPP430 or pMC73A was almost the same.

Objective To investigate the effect of sodium arsenite by Wnt signaling pathway on proliferation and apoptosis of oral squamous cell carcinoma.Methods Cell proliferation was detected after 1.25,2.5,5,10,20μmol/L sodium arsenite treatment human oral squamous cell carcinoma cell line Tca8113 for 24,48,72 hours by CCK8 experiment.0 and 14μmol/L sodium arsenite was used to treatment Tca8113 cells with 48h,cell apoptosis were detected by flow cytometry,Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by Western blot.Tca8113 cells were divided into control group,sodium arsenite group,activating agent+sodium arsenite group,all treated for 48hour,cell proliferation,apoptosis and Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by CCK8 assay,flow cytometry and Western blot.Results Tca8113 cell proliferation was inhibited significantly with the increase of treatment time and sodium arsenite concentration,and has a time and concentration dependent manner(P<0.05 or P<0.01).10μmol/L sodium arsenite as a follow-up study according to the IC50.Cell inhibition rate,apoptosis rate and Cleaved Caspase3 protein expression in 10μmol/L group were significantly higher than that of 0 mol/L group,the expression of β-catenin,Cyclin D1 protein was significantly lower than that of 0 mol/L group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in sodium arsenite group and activating agent+sodium arsenite group were significantly higher than control group,the expression of β-catenin and Cyclin D1 protein were significantly lower than control group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in activating agent + sodium arsenite group were significantly lower than that of sodium arsenite group,the expression of β-catenin and Cyclin D1 protein were significantly higher than that of sodium arsenite group(P<0.01).Conclusion Sodium arsenite can inhibit the proliferation of oral squamous cell carcinoma and promote apoptosis,and the mechanism was related to regulation of Wnt signaling pathway.

Objective To explore the correlation between hepatitis B virus large surface protein(HBV-LP)and hepatitis B virus deoxyribonucleic acid(HBV-DNA),analyze the clinical significance of detecting the two index dynamically for diagnosis and treatment of hepatitis B.Methods Enzyme linked immunosorbent assay(ELISA)and fluorescent quantitation polymerase chain reaction(FQ-PCR)were used to detect the levels of HBV-LP and HBV-DNA in serum specimen of 230 hepatitis B patients.Results There was a positive correlation between the content of HBV-LP and copy numbers of HBV-DNA in serum of hepatitis B positive cases(r=0.84,P＜0.01).The positive rate was 84.78％ in HBV-LP and 84.35％ in HBV-DNA of 230 HBV positive cases.As a result there was no significant difference(P＞0.05)in this study.Therefore,the positive rate was 63.63 ％(35/55)in HBV-LP and 58.18％(32/55)in HBV-DNA of 55 HBV negative cases.There was also no significant difference(P＞0.05)in this study.However,the positive rate of HBV-LP(82.47％)was higher than HBE(52.06％)in the 194 HBV-DNA positive cases.There was significantly different(P＜0.01).Conclusion HBV-LP and HBV-DNA are the significant index of the degree of hepatitis B virus replication in hepatitis B positive cases,especially in determining the virus replication and prognosis of treatment in HBe Ag negative,providing the reliable laboratory data for the clinical diagnosis and treatment.

Objective To investigate the impact of applying patient-centered care concept on easing the negative moods among the parents of the neonates hospitalized in NICU.Methods The parents of neonates (76 people) hospitalized in NICU from March to May in 2012 were selected as the control group,they conducted normal nursing.The parents of neonates hospitalized in NICU during June and August in 2012 were named as the observation group (81 people).In addition to the normal nursing,they were given nursing intervention according to patient-centered care concept.The anxiety and depression moods of the two groups was analyzed.Results Before the patient-centered care concept intervention,there was no statistical difference of SAS,SDS scores for both groups.For the observation group,the SAS,SDS scores after intervention were significantly lower than those before the intervention.While for the control group,there was no statistical difference of SAS,SDS scores.After intervention,the SAS、SDS scores of the observation group were significantly lower than those of the control group,and the difference was statistically significant.Conclusions The negative moods of the parents of hospitalized neonates can be eased significantly by applying patient-centered care concept intervention.