Objective To study the relation between the quantity of DNA extracted by Chelex-100 method and multiplex STRs analysis.Methods DNA extracted from a variety of common forensic casework specimens were quantified by using Real-time PCR,and then amplified with AmpFLSTR IdentifilerTM PCR Amplification kit.ResultsAccording to the results of quantification,the quantities of DNA extracted from 113 samples by Chelex-100 method were adjusted to 0.5~3ng for establishing 8?l amplification system,and in this condition,most of 113 forensic casework specimens could be successfully genotyped.Conclusion When the quantity of DNA extracted by Chelex-100 method ranged from 0.5ng to 3ng,most results of multiplex STRs analysis were satisfying.Moreover,the amplification effect of 1?l DNA template was better than 3?l DNA template when the concentrations of extracted DNA were more than 0.5ng/?l.

Objective To develop a rapid,sensitive,specific and accurate quantitative duplex real-time PCR assay for detection of Listeria monocytogenes and Shigella.Methods Two sets of specific primers and probes were selected according to Listeria monocytogencs hly gene and Shigella ipaH gene.The target hly and iPaH fragments were amplified by PCR,and used to construct recombinant pGEM-T-hly and pGEM-T-ipaH respectively.The two recombinant circular plasmid DNAs were linearized with EcoR I that did not cut within the target DNA fragment.The ten-fold dilutions of plasmid were subjected to the standard quantitation curve in duplex real-time PCR assay.Various genomic DNAs of Listeria innocua,Listeria weshimeri,Salmonella,Staphylococcus aureus,Bacillus subtilis,Escherichia coli and Proteus were used as negative controls to confirm the specificity of duplex real-time PCR assay.The assay was also used to detect Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk.Results The recombinant plasmids were constructed successfully,hly probe(rAM and TAMRA double labelled)and ipaH probe (HEX and TAMRA double labelled)were used to develop an optimized PCR successfuliv.Conclusion The selected primers and probes showed high specificity for these two target bacteria,the linear range of the assay was good(105-101 copies/μl,R2≥0.998)and sensitivity Was 10 copies/PCR.Following a DNA extraction method which combined EZ Spin Colum Genomic DNA Isolation Kit(BBI)/Phenol-chloroform,the sensitivity of assay Was 102CFU/ml for both Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk,which equivalents to 10 CFU/PCR.

AIM: To investigate the clinical therapeutic effects of human umbilical vein (HUV)implantation and mitomycin C (MMC) in non-penetrating trabecular surgery (NPTS). METHODS:A total of 32 patients (46 eyes) with uncontrolled primary open angle glaucoma (POAG) were divided into two groups: HUV+MMC group (n=25), SKGEL+MMC group (n=21). The procedure commenced with the creation of a limbus based conjunctival flap. After the dissection of a superficial limbus based rectangular scleral flap, MMC(0.4mg/mL) was used superior and inferior surface of the superficial scleral flap for three minutes. A second limbus based scleral flap was carefully dissected beneath the previous one towards the choroid. Schlemm's canal was deroofed during the extension of the deep scleral flap toits limbal edges. HUV or SKGEL fixed on the bed of sclera in experimental group. Postoperative examinations were performed at 1 week,2,4 weeks;2,6,12 months. IOP,best-corrected visual acuity(BCVA), functional blebs and success rate were examined. RESULTS: There were no statistically differences with postoperative IOP in HUV+MMC group and SKGEL+MMC group (P>0.05) during 1 week to 12 months. There was no difference with postoperative function blebs and the change of BCVA during 1 week to 12 months between HUV+MMC group and SKGEL+MMC group (P>0.05).At 12 months after surgery, the success rate was 84% in HUV+MMC group,86% in SKGEL+MMC group. CONCLUSION: The application of HUV in NPTS can prevent the adhesion of filtering channel and it can improve the success rate of NPTS. Compared with SKGEL, HUV has lower price. So it is a better implant.

Objective To study the application of a new magnetic beads made in China in DNA extraction of forensic biological samples with automation workstation.Methods DNA was extracted from common forensic biological samples by QIAGEN Bio-Robert Universal System and a new magnetic beads made in China,and then typed with Identifiler system in ABI3130XL Genetic Analyzer.210 of these samples were also quantitated by ABI7500 Real Time System.Results Total of 9100 genomic DNA was extracted from various forensic biological samples by the new magnetic beads made in China and automation workstation methods,and most of them were successfully typed for STR analysis.In these biological samples,oral swabs and muscles were of the highest Success rate of STR typing(100%),and the lowest was touched cell samples (50.0%).Conclusion The new magnetic beads made in China with automation workstation methods can be applied to DNA extraction of most forensic biological samples.

Objective To observe the anti-influenza effect of a Chinese medicinal herb-Antiviral Agent No.1. Methods To study t he anti-viral effect of Antiviral Agent No.1 by means of the technique of cel l culture and using Ribavirin as a positive control. Results In MDC K cu lture, Antiviral Agent No.1 was found to be a potential inhibitor of influenza A 3 virus in a concentration-dependent manner, with a TC50 of 60.53 mg*m l-1. When drug was added 2 hours post virus infection, the EC50(TI) was 5.14 mg*ml-1(11.78); while drug was added 2 hours before infection, t he EC50(TI) was 5.20 mg*ml-1(11.65). Conclusions Antiv iral Agent No.1 had a significant anti-influenza effect on type A3 in MDCK.

Actins ; metabolism ; Bone Neoplasms ; metabolism ; pathology ; surgery ; Desmin ; metabolism ; Diagnosis, Differential ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Leiomyosarcoma ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Neurilemmoma ; metabolism ; pathology ; Tibia ; Vimentin ; metabolism

BACKGROUND:Erythromycin spreads widely in the body with a short period of effective concentrations and has a lot of adverse effects.Therefore,it is necessary to make erythromycin as targeted medicine.OBJECTIVE:To optimize the preparation conditions of erythromycin gelatin microspheres.DESIGN,TIME AND SETTING:An orthogonal controlled test was performed in the Department of Pharmacy,Guangdong Pharmaceutical University from June to December in 2005.MATERIALS:Erythromycin and gelatin.METHODS:According to the emulsion principle,erythromycin dispersed in the gelatin solution.In the process of preparing microspheres,the gelatin solution and oil should form W/O emulsion and then it turned into spheres by solidification.The formation and quality of microspheres were influenced by four factors,namely the concentration of gelatin,dosage of emulsifier,the solidification time and the speed of mixing.The arithmetic mean diameter of microspheres,the drug loading efficiency and the encapsulation efficiency were targets for the survey in this study on the basis of pretests.The best preparation conditions were optimized in accordance with the results of L9 (34) orthogonal tests.The optimized preparation conditions were obtained according to the results of orthogonal tests.MAIN OUTCOME MEASURES:The mean diameter of microspheres,the drug loading efficiency,the encapsulation efficiency,and the orthogonal tests were examined.RESULTS:The optimized preparation conditions of erythromycin gelatin microspheres included 15% gelatin,3.0 mL emulsifier,0.5 hour solidification and mixing at 1 000 r/rain.The erythromycin gelatin microspheres were regular in their morphology.Drug was enveloped in microspheres.The average particle size was (14.15±0.20) μm;the drug loading efficiency and the encapsulation efficiency were (5.83±0.38)% and (65.70±0.56)%,respectively.Over 90.16% of the microspheres was in the range of 7-25 μm;The reappearance of pharmaceutical technology was good.CONCLUSION:The optimized preparation conditions of erythromycin gelatin microspheres are obtained using L9 (34)orthogonal tests.The microspheres prepared meet the requirement of the size for lung targeting.

Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

Objective:To study the relationship among lipoprotein‐associated phospholipase A2 (Lp‐PLA2 ) gene A379V and T403V locus polymorphisms and genetic susceptibility of coronary heart disease (CHD) .Methods:Lp‐PLA2 gene A379V and T403V locus polymorphisms of 160 coronary angiography confirmed CHD patients (CHD group ) and 117 healthy subjects (healthy control group ) were measured using gene sequencing technique .ELISA was used to measure blood lipids and plasma Lp‐PLA2 level in two groups ,and they were compared between two groups . Results:Compared with healthy control group ,there were significant rise in age ,male proportion ,plasma levels of hs‐cTnI ,hsCRP ,TC ,LDL‐C , Lp (a) ,WBC ,mononuclear cells (MNCs) and Lp‐PLA2 [ (119.98 ± 49.41) ng/ml vs .(248.59 ± 76.51) ng/ml] ,and significant reduction in HDL‐C level in CHD group ( P<0.01 all) .The CC , CT , TT genotype and C , T allele were de‐tected all in A379V and T403C locus of two groups .Compared with healthy control group ,there were significant rise in frequencies of CC genotype (1.7% vs .9.3% ) and C allele (13.7% vs .20.3% ) of Lp‐PLA2 gene T403C locus in CHD group , P< 0.05 both . All genotypes and alleles of A379V locus possessed no significant difference between CHD and healthy control group . Conclusion:Plasma Lp‐PLA2 level may be related to CHD risk .Lp‐PLA2 gene T403C locus poly‐morphism possesses certain relationship with genetic susceptibility of CHD .

Background Neovascular glaucoma is a type of refractory glaucoma.Biological amnion combined with glaucoma valve implantation is a primary therapy and its long-term effectiveness is noticeable. Objective The goal of this Survey was to evaluate the effectiveness of biological amnion combined with glaucoma valve implantation for neovascular glaucoma and compare the clinical outcome with simple glaucoma valve implantation. Methods This was a retrospective observational case series.The clinical data of 44 eyes of 44 patients received biological amnion combined with glaucoma valve implantation for neovascular glaucoma and 43 eyes of 43 patients received simple glaucoma valve implantation for neovascular glaucoma were retrospectively analyzed and compared.The age,sex and disease-cause were matched between these two groups.Patients were followed-up for 24 months after operation.Surgery success was identified as the intraocular pressure(IOP)<21 mmHg after operation.Written informed consent was obtained from each patient prior to the operation.Results The IOP was<21 mmHg throughout the follow-up duration in both groups.No significant difierence was found in the IOP value in 1 week after operation between two groups(t=-5.34,P=0.60).However,IOP values were lower in biological amnion combined with glaucoma valve implantation group in 3,12 and 24 months after operation than those of simple glaucoma valve implantation(t=6.64,t=5.00,t=7.81,P<0.01).Operation successful rates in biological amnion combined with glaucoma valve implantation group were 97.73％.93.18％。90.24％and 82.05％in 1 week,3 months,12 months and 24 months respectively after operation.and those in simple glaucoma valve implantation were 95.35％,71.43％,65.00％and 60.53％in corresponding time points,showing considerably significant differences between two groups (χ2=7.06,χ2=7.47,χ2=4.37,P<0.05).There was no significant difference in Ihe number of eyes with complication between the two groups(P>0.05). Conclusion The biological amnion combined with glaucoma valve implantation surgery may be more effective and safe for the treatment of neovascular glaucoma than with glaucoma va]ve only.