It is well documented that altered biosynthesis of cell surface N-linked oligosaccharides is associated with the transformed cells and tumors.N-acetylglucosaminyltransferase Ⅱ(GnT-II;EC 2.4.1.143)is a medial Golgi enzyme that catalyses the incorporation of a GlcNAc residue in ?-1,2 linkage to the Man-?-1,6 arm of the N-glycan core.This is an essential step in the biosynthetic pathway leading from hybrid to complex N-glycans.Because functional GnT-Ⅱ is an prerequisite of N-Acetylglucosaminyltransferase V performance,It was speculated that GnT-Ⅱ was involved in cancer development and progression.The expression of GnT-Ⅱ in mouse breast cancer cells 67NR and 4T1 which have different behavior of metastasis was analysed using RT-PCR.The amounts of GnT-Ⅱ in the highly metastatic cell 4T1 increased to 1.53 times of the lowly metastatic cell 67NR.To determine the association of GnT-Ⅱ with tumor progression,the GnT-Ⅱ encoding gene was amplified with RT-PCR and cloned into retrovirus vector pMSCV,resulting in pMSCV-GnT-Ⅱ.The recombinant plasmid was transfected into 4T1 and the transfected cells were selected in the medium containing puromycin,which were harvested to detect the adhesion ability to fibronection and the migration potential by transwell system.The cell adhesion to fibronectin was weakened by 67% and migration potential was increased by 82%.The data indicates that GnT-Ⅱ mediates cell adhesion and migration,thus may play an important role in cancer metastasis.

Objective To investigate the oxalate-degrading and growth abilities of lactic acid bacteria coming from yoghourt in vitro culture broth. Methods The species of lactic acid bacteria obtained from 6 brands of yoghourt were identified. Different brands of yoghourt were separately cultured in broth and incubated at 37 ℃,5% CO2 for 48 h, getting the concentration of lactic acid bacteria through slab counting method. The brand of yoghourt with the highest quantity of bacteria was chosen as the study subject. Three species of bacteria and the mixture bacteria obtained from the chosen brand of yoghourt were cultured separately for 72 h in MRS broth which contained three different concentrations of ammonium oxalate, and the broth without inoculating of bacterium was prepared as control. The amount of bacteria in culture was detected, the concentration of ammonium oxalate was detected through photometric determination of oxalic acid by its catalytic reaction of methyl red by potassium chromate. Results Three species of lactic acid bacteria (L.bulgaria, L.acidophilus and L.thermophilus) were detected in each brand of yoghourt. The quantity of bacteria in Weiquan yoghourt is the highest. After 72 h incubation, the concentrations of oxalate in all the broths containing bacteria were lower than those in the control. L.acidophilus showed the best degrading activity. The higher the concentration of oxalate was, the bigger the amount of oxalate degraded was, the smaller the percent of degrading was, and the less the growth ability of bacteria was. Conclusion All the species coming from yoghourt have the ability of oxalate degrading, which increases with the concentration of oxalate in culture.But the high concentration of oxalate restrains the growth of bacteria.

AIM:To report the safty and efficiency of intravitreal injection of bevacizumab (Avastin) in patients with macular edema (ME) due to branch retinal vein occlusion (BRVO).METHODS: A consecutive series of patients with ME due to BRVO who were treated with intravitreal bevacizumab injection (2.5g/0.1L) were retrospectively studied. Patients underwent complete ophthalmoscopic examination, including Snellen visual acuity testing, optical coherence tomography (OCT) imaging, and/or flurescence angiographic testing at baseline and follow-up visits.RESULTS: There were 32 eyes of 32 consecutive patients who received at least one intravitreal bevacizumab injections (range from 1 to 3). The mean length of follow-up was 4.7 (range from 3 to 8) months. The mean visual acuity improved from 20/200- at baseline to 20/100- at 1 month and 20/100+ at 3 months and last follow-up (P＜0.01). The mean central 1mm macular thickness was 483μm at baseline and decreased to 275, 314,and 301μm at 1 month,3 months, and last follow-up (P＜0.01)respectively.No adverse side effects were observed following injections in any eyes.CONCLUSION: Intravitreal bevacizumab (Avastin) showed a marked decrease in ME secondary to BRVO, improvement in visual acuity and lack of adverse side effects.

Objective · To investigate effect of fucoidan on autophagy, migration and invasion in human multiple myeloma U266 cells. Methods · The U266 cells treated with fucoidan were cultured in vitro. The formation of autophagosomes was observed by transmission electron microscopy (TEM). Transwell assay was used to evaluate the effect of fucoidan on migratory and invasive abilities of U266 cells. The protein levels of LC3-Ⅱ/LC3-Ⅰ, Beclin-1, P62, MMP9, CXCR4, p-AKT/T-AKT, p-mTOR/T-mTOR were detected by Western blotting. MMP9 concentration in the culture medium was examined by ELISA. Results · ① Autophagosomes increased in fucoidan-treated cells compared with control group under TEM. ② Migratory and invasive abilities were inhibited by fucoidan in a dose-dependent manner, which were suppressed by chloroquine. ③ Western blotting demonstrated that expression of LC3-Ⅱ/LC3-Ⅰ, Beclin-1 and MMP9 increased in fucoidan-treated cells, while P62, CXCR4, p-AKT/T-AKT and p-mTOR/T-mTOR decreased compared with control group. ④ The result of ELISA showed that MMP9 concentration in the culture medium of fucoidan-treated cells significantly decreased. Conclusion · Fucoidan induces autophagy and inhibits migration and invasion in U266 cells.

Objective To investigate the prevalence of integrons in A.baumannii isolates,analyze the correlation between inte- grons and resistance of A.baumannii,and study the resistance genes in integrons.Methods A total of 106 strains of A.bau- mannii were collected to test the antibiotic susceptibility by disk diffusion method.The classification of integrons was per- formed by analyzing the positive PCR products using restriction fragment length polymorphism (RFLP).The variable region of integrons was amplified by integron PCR.RFLP and DNA sequencing were used to analyze the resistance genes in integrons. Results About 52.8% (56/106) of the isolates showed integron positive.PCR-RFLP analysis revealed that they were all class I integrons.About 94.6%(53/56) of the positive strains with integrons owned the variable region,which was confirmed by integron PCR.The sizes of the amplicons ranged from 0.15 kb to 2.8 kb.All together 7 different cassette arrays were detec- ted,including genes coding resistance to aminoglycosides (aadA1,aadA2,aadA5,aadB,aacA4),sulphonamides (dfrⅫ, dfr17),?-lactam compounds (bla_(ara-10)),chloramphenicol (catB-like,catB8),and two open reading frames (orfF,orfI) with unknown function.A novel cassette array orfI-aadA1 was reported,and its GenBank accession number was DQ092497.Conclu- sions Class I integrons are widespread in A.baumannii isolates in Nanjing.The integrons are closely associated with the resist- ance and multidrug resistance in A.baumannii isolates.

OBJECTIVETo investigate the effect of dihydrotestosterone (DHT) on the gene transcriptions and expressions of Smad3 and Smad4 in androgen dependent prostate cancer cell line LNCaP, and whether this effect can be suppressed by the androgen receptor inhibitor flutamide.

METHODSThe androgen dependent prostate cancer cell line LNCaP was cultured in RPMI 1640 medium and treated with different concentrations of DHT(2, 10, 50 nmol/L) and flutamide (100 nmol/L). Quantitative reverse transcription PCR (RT-PCR) was used to detect the mRNAs of Smad3 and Smad4. The expressions of Smad3 and Smad4 protein were detected by Western blot assay.

RESULTSCompared with the control group without any DHT or flutamide, higher concentration(10, 50 nmol/L) of DHT enhanced the transcription of Smad3 mRNA (P <0.05). Serial concentrations of DHT increased the expression of Smad3 protein(P < 0.05). Flutamide inhibited the up-regulation of both Smad3 mRNA transcription and expression significantly (P <0.05). 10 nmol/L DHT significantly suppressed the transcription of Smad4 (P <0.05). There was considerable suppressions of Smad4 expression at the presence of DHT in different concentrations (P < 0.05). And the degree of this suppression was more significant than that of DHT on Smad4 mRNA transcription. Flutamide inhibited the suppressive effects of DHT on both Smad4 mRNA transcription and expression.

CONCLUSIONDHT can enhance the transcription and expression of Smad3, while it decreases the transcription and expression of Smad4 in LNCaP cell line. There is a possible crosstalk between the AR signal and TGF-beta signal passways at the level of Smads.

Androgens ; physiology ; Cell Line, Tumor ; Dihydrotestosterone ; pharmacology ; Flutamide ; pharmacology ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; Smad3 Protein ; biosynthesis ; genetics ; Smad4 Protein ; biosynthesis ; genetics ; Transcription, Genetic

OBJECTIVE: To investigate the effect of different PCR amplification volume on the accuracy of human identification test. METHODS: Human genome DNA samples were amplified using ABI PRISM Profiler Plus kits in 50 microliters, 25 microliters, 12.5 microliters, and 6.25 microliters reaction volume, respectively. The thermocycle parameters were the same. All PCR products were then electrophoresized on ABI PRISM 310 Genetic Analyzer, 377 DNA Sequencer, and 3100 Genetic Analyzer. Data were processed by ABI PRISM GeneScan and Genotyper software. RESULTS: The less reaction volume, the more alleles losing or alleles adding observed. CONCLUSION Non-standard volume of PCR amplification reaction should be used carefully in human identification test, especially when the sample DNA quality is not so satisfied.
Alleles ; Blood Stains ; DNA/isolation & purification* ; Electrophoresis ; Forensic Medicine ; Humans ; Loss of Heterozygosity ; Polymerase Chain Reaction/methods* ; Sensitivity and Specificity ; Spectrophotometry, Ultraviolet

OBJECTIVE: To explore the feasibility of detecting of Y-STR of fetal DNA in maternal plasma using Ion Torrent PGM™ System. METHODS: A total of 16 fetal DNA samples from maternal plasmas (8 cases from 38 weeks gestational age and 8 ones from 12 weeks) were prepared and a multiplex assay with 7 STR loci (DYS390, DYS391, DYS393, DYS438, DYS437, DYS456, DYS635) was designed for multiplex-PCR amplification. Using Ion Torrent PGM™ System, the results of Y-STR sequences and capillary electrophoresis were obtained and compared. RESULTS: Y-STR specific alleles were detected in the maternal plasma of all the pregnant women having male babies of second and third trimester, which were higher than that detected by capillary electrophoresis. Consistent Y-STR genotypes were observed between fetal DNA from maternal plasma and genomic DNA from the newborn babies. CONCLUSION Based on Ion Torrent PGM™ System, the prenatal Y-STR detection method may provide a high-sensitive and high-throughput choice for prenatal STR detection in forensic testing.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA/blood* ; Family ; Female ; Fetal Blood/chemistry* ; Genotype ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pregnancy ; Sensitivity and Specificity ; Sex Determination Analysis ; Tandem Repeat Sequences/genetics*

Objective To observe the therapeutic effect of therapy of tonifying kidney,activating bone-marrow, and unblocking collaterals for patients with chronic aplastic anemia (CAA), and to investigate its effect on thromboelastogram platelet maximum amplitude (Ma) value for exlporing its therapeutic mechanism. Methods Sixty CAA patients were randomized into trial group and control group, 30 cases in each group. The control group was given oral use of Stanozolol and Cyclosporin A, and the trial group was orally given the recipe with the actions of tonifying kidney,activating bone-marrow,and unblocking collaterals,which is mainly composed of Radix Rehmanniae,Radix Rehmanniae Preparata,Caulis Spatholobi,Semen Cuscutae,Fructus Lycii,Radix Angelicae Sinensis, Fructus Ligustri Lucidi, Herba Ecliptae, Pheretima, and Semen Strychni Preparata. The clinical efficacy was evaluated after treatment,and peripheral hemogram and thromboelastogram Ma value of the two groups were compared before and after treatment. Results (1)The trial group had better western medicine therapeutic effect and traditional Chinese medicine (TCM)syndrome therapeutic effect than the control group, the difference being signficant (P < 0.01).(2) After treatment, TCM syndrome scores, parameters of blood routine test,thromboelastogram Ma value of the two groups were improved compared with those before treatment (P < 0.05 or P < 0.01),and the improvement in the trial group was superior to that in the control group (P <0.05). Conclusion Therapy of tonifying kidney, activating bone-marrow, and unblocking collaterals is effective on improving blood coagulation function by increasing the quality and amount of platelet.

Background Corneal neovascularization ( CNV ) is a complication of many ocular surface diseases.It often worsen the pathological course.Effective therapy for CNV is still researching. Objective This study was to investigate the inhibitory effect of irradiation on CNV. Methods CNV models were established in 70 right eyes of 70 clean Wistar rats by corneal alkali burning.The models were randomized into β ray 10 Gy once irradiation group( 2 eyes),β ray 7 Gy multiple irradiation group( 17 eyes),β ray 10 Gy multiple irradiation group( 17 eyes),1％ cyclosporin A ( CsA ) eye drops group ( 17 eyes) and model group ( 17 eyes),and 6 matched normal rats were used as normal controls.All treatments started from the first day of the corneal alkali burning.CNV length and area were measured under the slit lamp every day.Corneal samples and homogenate were prepared 3,5,7 days after corneal alkali burning.The expressions of bcl-2,bax,vascular endothelial growth factor(VEGF) in rat corneas were detected by immunochemistry,VEGF proteins and VEGF mRNA were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR),respectively. Results Corneal ulceration was found in the βray 10 Gy once irradiation group and β ray 10 Gy multiple irradiation group.CNV length and area were much less in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group compared with the model group on the seventh day after experiment( length:q=14.40,24.20,P＜0.01 ;area:q=17.80,14.00,P＜0.01 ).Immunochemistry revealed that compared with the model group,expressions of bcl-2 and VEGF proteins were weaker,but the expression of bax protein was stronger in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group.RT-PCR showed that the expression of VEGF mRNA in cornea was lower in the β ray 10 Gy multiple irradiation group,β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group in comparison with that in model group,and the results from Western blot showed the same pattern as RT-PCR. Conclusions Low dose irradiation of 90Sr-90Y ophthalmic applicator inhibits CNV formation after alkali burn.The study provide a new understanding of the irradiation for the treatment of CNV.