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.

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 .

·AIM: To study the expressive variation of Nogo-A on rat retina in the process of chronic ocular hypertension. · METHODS: Thirty-six healthy adult male Wistars were randomly divided into control group (6 rats) and chronic hypertension group (30 rats). Chronic hypertension was created by cauterizing the superficial scleral veins. Immunohistochemistry technique was used to evaluate the expressive varieties of Nogo-A at different time points during the course of chronic ocular hypertension. · RESULTS: The success of the model was indicated by over 40% of increase in the IOP as compared with normal rats. Compared with control group, as time passed chronic hypertension group gradually had detectable morphology changes in the retina. At the 21st day of chronic ocular hypertension, retinas became thinner and the quantity of retinal ganglion cell (RGC) decreased (P<0.05). Assoicated with the morphological changes, the expression of Nogo-A was strongly increased (P<0.05).CONCLUSION: Myelin associated protein Nogo-A plays a part in the process of chronic ocular hypertension.

AIM:To compare one-site vs two-site phacotrabe-culectomy in chronic angle-closure glaucoma (CACG) coexisting with cataract.METHODS:This prospective, randomized study included 41 eyes with CACG. One-site approach was performed in 21 eyes and two-site procedure in 20 eyes. Intraocular pressure (IOP), best-corrected visual acuity (BCVA), the number of antiglaucoma medications and complications were observed. All patients were followed up for 9 months.RESULTS:There were no significant differences between the two groups preoperatively. IOP decreased from 22.7±4.9mmHg and 23.7±4.7mmHg preoperatively in one-and two-site groups to 18.0±1.2mmHg and 16.7±1.1mmHg 9 months after operation respectively(P<0.05). There were no significant differences in mean IOP between the two groups at any time (P>0.05). Decrease of the number of antiglaucoma medications and BCVA improvement were similar in both groups 9 months after surgery (P>0.05).There were no significant differences in complications between the two surgical procedures.CONCLUSION:There were no significant differences between the two groups in clinical efficacy and complications.