ObjectiveTo test the expression of epidermal growth factor-like domain 7 (EGFL7),microvessel density (MVD) and foeal adhesion kinase pY397 (FAKpY397) in human glioma tissues,and to evaluate their relationship.MethodsThe expression of EGFL7 and FAKpY397 in 56 cases of human glioma and 8 cases of normal brain tissues were detected by immunohistochemistry test,and MVD was detected by CD34 staining.ResultsThere was a significant difference of the positive rates of EGFL7 between normal brain tissue (0) and gliomas (75％),χ2 =17.45,P ＜0.01.With the increased pathological grade,the expression level of EGFL7 increased (χ2 =26.24,P ＜ 0.01 ).There was a significant difference of the positive rates of FAKpY397 between normal brain tissue ( 12.5％ ) and gliomas (73.2％),χ2 =6.23,P ＜ 0.05.With the increased pathological grade,the expression level of FAKpY397 increased (χ2 =6.71,P ＜ 0.01 ). MVD on normal brain was( 15 ± 4 )/HP,on Ⅰ - Ⅱ grade and Ⅲ -Ⅳ grade gliomas was ( 27 ± 3 )/HPand ( 60 ± 4 )/HP respectively,there was a significant difference on MVD between normal brain tissue and gliomas (P ＜ 0.01 ).Higher level of MVD was found in gliomas with higher grade ( P ＜ 0.01 ).There was a positive correlation between EGFL7 and FAKpY397 expressions in gliomas (r =0.314,P ＜0.01 ).There was a significant difference on MVD between positive and negative expression of EGFL7 ( t =26.55,P ＜ 0.01 ). MVD was (56 ± 4 )/HP and (25 ± 3 )/HP respectively.ConclusionThe expression of EGFL7 of human gliomas has a favorable positive correlation with the degree of malignancy,MVD and FAKpY397.It is indicated that EGFL7not only palys an important regulative role in glioma neovascularization,but also it may participate directly in glioma occurrence and invasion.

Objective To explore the efficacy of intraductal chilled saline perfusion (ICSP) to reduce the thermal bile duct injury during the treatment of radiofrequency ablation (RFA) associating with occlusion of hepatic blood supply in rabbits.Methods 16 healthy New Zealand white rabbits were divided into 2 groups.Rabbits of the ICSP group were placed tubes in the common bile duct after laparotomy,and ICSP was performed during the RFA procedure accompanied with hepatic blood occlusion.While for rabbits of the non-ICSP group,hepatic blood occlusion and RFA were performed without ICSP.RFA electrodes were placed about 5 mm away from the hilus hepatis approximately.Contrast-enhanced ultrasonography (CEUS) was administrated to evaluate the sizes of the ablative zones after the procedure.On post-procedure 6 week,ultrasonography was prerformed to evaluate the changes of the biliary structure,and liver specimens of rabbits wcrc obtained for histopathologic observation of main bile ducts.Results Post-procedure CEUS examination showed that there was no significant difference in the size of the ablative zone between the groups (P ＞0.05).On post-procedure 6 week,rabbits of the ICSP group appeared with biliary dilatation more frequently by ultrasonography (P ＜0.05),and a higher degree of the injury of main bile duct by histopathologic observation (P ＜0.05).Conclusions In treatment of RFA accompanied with hepatic blood occlusion,RFA-induced bile duct injury may be decreased significantly with ICSP.

There is no defined legislation on paternity testing in present China,thus the testing status quo is in a mess to some extent.Together with other unfavorable factors including the unstable marital status in modern society,increasing extramarital sexual behaviors and illegitimate children,and the ever-heated testing competition among testing institutes driven by the economic interests and so forth,the paternity testing is on a sharp rise these years,which leads to a increasing chaos in paternity testing field.Social ethical crisis is partially responsible for the current problem,thus related ethical issues to paternity testing are discussed in this article.

Objective To analyze the characteristics of blood loss for laboratory test of critically ill premature infants,and to seek feasible measures to reduce the blood loss.Methods Two hundred and forty-six cases of critically ill premature infants admitted to the neonatal intensive care unit from April 2012 to April 2013 were analyzed the blood loss for test during the hospitalization and the clinical features of blood loss with different gestational age,different weight within the first four weeks after admission.Then the application of blood loss according to test category was described.The blood volume demanded in theory was determined by the formula B =5 (∑ s + 0.1),then calculated the phlebotomy overdraw on the basis of the practical blood loss and analyzed the characteristics of overdraw per patients per day in first two weeks after admission.Results Among 246 patients,The median blood loss figure was 25.57 ml for each infants with the range between 7.10 ml ～ 119.20 ml,and the blood loss concentrated in first four weeks,which showed a decreasing trend with time.There was a statistically significant difference(P ＜0.05)that the smaller gestational age,the lower birth weight,the more daily blood loss per patient per day in first two weeks,but no significant differences(P ＞ 0.05) between the third and fourth week.The largest proportion of the blood samples was used for clinical chemical tests(31.49％),followed by blood gas analysis (19.03％),immunoassays (12.69％),blood cultures (12.63％),hematology (12.28％).The practical blood loss was about twice times of blood volume demanded for tests in theory,which the median was 7.8 times to the latter(25.57 ml vs 3.26 ml).It showed statistically significant difference(P ＜0.05) between blood overdraw per patients per day in the first week and the second week.Compared with different gestational age and birth weight,the difference of overdraw was also statistically significant(P ＜0.05).According to test category,blood culture was the most significant samples of phlebotomy overdraw,followed by biochemical,other,blood gas analysis,the percentage was 76％,64％,45％ and 41％ respectively.Conclusion The blood loss for laboratory test and the phenomenon of blood waste is serious in critically ill preterm infants.The smaller the gestational age is,the lower the weight is,the amount of blood loss and phlebotomy overdraw are more significantly.Biochemical and blood gas analysis are the main items of blood loss.

Objective:To evaluate the measurement uncertainty in the determination of sodium valproate tablets by GC with an in -ternal standard method , and determine the main sources of uncertainty .Methods:A GC internal standard method was selected to sys-tematically analyze the uncertainty in the determination of sodium valproate tablets , including the sample quantity , dilution ratio, purity and area repeatability of chromatographic peaks .Results: The expanded uncertainty of sodium valproate tablets was 2.7%, and the determination range of sodium valproate tablets was (96.3 ±2.7)%(k=2).Conclusion:The established GC internal standard meth-od for the uncertainty evaluation is reliable , which is helpful to improving the quality evaluation and control of sodium valproate tablets .

OBJECTIVE:To investigate the similarity of dissolution curves between generic preparations and reference prepara-tions of Bisacodyl enteric-coated tablets in various dissolution mediums,and to provide reference for improving production technolo-gy and internal quality of generic preparations. METHODS:Paddle method was adopted with rotation speed of 75 r/min. The disso-lution test was performed using 1000 mL pH 6.0 phosphate buffer solution,pH 6.8 phosphate buffer solution,water containing 2% sodium lauryl sulfate. HPLC method was used to determine average accumulative dissolution of main components from 3 kinds of generic preparations and reference preparations at different time points to draw out dissolution curves. Similarity factor(f2)meth-od was used to the similarity of dissolution curves. RESULTS:Dissolution curves of reference preparations were basically the same in 3 kinds of dissolution mediums. But the dissolution curve f2 of one generic preparation among 3 manufactures to dissolution curve of reference preparation were ≥50,namely the similarity. CONCLUSIONS:The quality of generic Bisacodyl enteric-coat-ed tablets produced by different manufacturers is obviously different;the generic preparations needs to be further improved in the production technology and internal quality. For domestic generic preparation,it is necessary to strengthen the real-time monitoring of its dissolution curve,to ensure the drug quality.

OBJECTIVE:To establish the purity determination of bisacodyl by differential scanning calorimetry(DSC)and the valuation of uncertainty. METHODS:DSC was conducted to detect the purity of bisacodyl and determine the optimal testing condi-tions. According to related standards,indium enthalpy change values,measurement repeatability,weighing process,instrument tem-perature deviation and system software deviation were systematically analyzed. The results were verified by HPLC. RESULTS:When the fiducial probability P was 0.95,the standard value and uncertainty of content of bisacodyl was (99.88 ± 0.06)% mea-sured by DSC. Weighing process,instrument temperature deviation and system software deviation had great effects on the total un-certainty. The result of HPLC and DSC were the same. CONCLUSIONS:The established DSC can quickly and accurately determine the chemical purity of bisacodyl. The uncertainty evaluation is reliable. Regularly calibrated and verificated equipment and strict con-trol of the weighing process will help to improve the accuracy measured by DSC;and it provides a new analysis method for the de-termination of purity of bisacodyl.

OBJECTIVE:To establish a method for simultaneous residual determination of dichloromethane and ethyl acetate in bisacodyl raw material. METHODS:Head-space GC was performed on the capillary column of 6% cyanopropyl phenyl-94% di-methyl polysiloxane(DB-624)by temperature programming,the temperature of injector was 220 ℃,detector was flame ionization detector with temperature of 250 ℃,carrier gas was high purity nitrogen with the flow rate of 3.0 ml/min,split ration was 1∶10, headspace heating temperature was 70 ℃,equilibration time was 30 min,volume of headspace vial was 5 ml,and the injection volume was 1 ml. RESULTS:The linear range was 6-120μg/ml for dichloromethane(r=0.999 9)and 50-1 000μg/ml for ethyl ac-etate(r=0.999 9);the limit of quantitation was 0.2,1.7 μg,limit of detection was 0.06,0.5 μg;RSDs of precision,stability and reproducibility tests were no higher than 3%;recoveries were 100.30%-102.00%(RSD=0.63%,n=9) and 100.10 %-101.30%(RSD=0.44%,n=9). CONCLUSIONS:The method is simple and accurate,and can be used for the simultaneous residual deter-mination of dichloromethane and ethyl acetate in bisacodyl raw material.

OBJECTIVE:To establish a method for the determination of content and content uniformity in Bisacodyl enter-ic-coated tablet. METHODS:HPLC method was performed on the column of Agilent ZORBAX C18 with mobile phase of acetoni-trile-20 mmol/L ammonium acetate(adjusted pH to 5.0 with acetic acid)(55∶45,V/V),the detection wavelength was 265 nm,col-umn temperature was 30℃,flow rate was 1.0 ml/min,and the volume injection was 20 μl. RESULTS:The linear range of bisaco-dyl was 50-1 000 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 99.50%-101.17%(RSD=0.5%,n=9). CONCLUSIONS:The method is reproducible with high accuracy,and suitable for the quali-ty control of Bisacodyl enteric-coated tablet.

Objective: To establish a GC method for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.Methods: The sample was dissolved in water, alkalified by sodium hydroxide solution and extracted by methylene chloride.An HP-5 gas chromatography column (50 m×0.32 mm, 1.05 μm) was used.The column temperature was programming increased, and the initial temperature maintained at 120 ℃ for 3 min, and then raised to 220 ℃ at a rate of 10℃·min-1 and maintained for 7 min.A hydrogen flame ionization detector (FID) was used and the split ratio was 1∶1.The inlet temperature was 230 ℃ and the detector temperature was 260 ℃.The injection volume was 1 μl and the carrier gas was nitrogen with high purity at a flow rate of 3.0 ml·min-1.Adamantane was used as the internal standard, and the internal standard method was used for the calculation.Results: The calibration curve was linear over the range of 0.05-1.0 mg·ml -1 (r=0.999 7).The detection limit and the limit of quantification was 1.1 ng and 3.3 ng, respectively.The average recovery was 100.2% (RSD =0.73%, n=9).Conclusion: The method has the advantages of simple operation, small extraction process toxicity, little environmental pollution, high accuracy and high specificity, and can be used for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.