Background The retina microglia play a eliminating effect on apoptotie cells in the neural retinal layer of normal rats during postnatal development.Milk fat globule epidermal growth factor 8(MFG.E8)can combine specifically with phosphatidylinositol serine of the surface of apoptotie cells and enhance macrophage phagoeytosis of apoptotic cells.Objective Present study was to evaluate the localization and expression of MFG-E8 and its relevant cytokines in the neural retinal layer of normal rats during postnatal development Methods Normal royal college of surgeon(RCS)rats were divided into P0,P3,P7,Pi4,P30,P45 groups according to their postnatal days,and the 30-day-old RCS rats(2 rats)served as controls.Double stain of M FG.E8 and microglial cells marker(CD11b)was performed by immunofluorescence.Expressions of MFG-E8,integrin β5,CD11b and interleukin-6(IL-6)mRNA in the neural retina were analyzed by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).The utilization of animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State and Science and Technology Commission.Results MFG-E8 and CD11b were positively co-expressed in retinal ganglion cell layer and external plexiform layer with the green fluorescence for FITC-labeled IgG and red fluorescence for cy3-labeled lgG respectively in normal adult rats.RT-PCR showed that the mRNA of MFG-E8,integrin 85,CD11b and IL-6 was detectable at P0 rats.The expression level of these eytokines began to rise fterward and reached peak value at P14 rats and then declined gradually,showing significant differences among different ages groups in various cytokines mRNA expression(all P<0.05).Conclusion MFG-E8 can be specifically expressed in the neural layer of retina microglia in RCS rat.

In order to obtain higher L-lactic acid yield industrial strain, the original strain Rhizopus oryzae PW352 was mutated by means of N+ ions implantation and a mutant strain Rhizopus oryzae RE3303 was obtained. Its lactic acid yield was increased by 75% than that of the original one. The acid producing condition was optimized by orthogonal design. The concentration of L-lactic acid reached to 131～136g/L and the conversion rate of glucose was as high as 86%～90% under the optimum condition.

One alkaline protease from Actinomucor elegans AS3.2778 was purified protein. The enzyme was purified using ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and size exclusion chromatography method, and its properties were also investigated. The molecular weight of this enzyme is 32 kDa with SDS-PAGE method, optimum temperature is 60℃, optimum pH is 8.5 to 10.5, it is stable in the pH range of 6.0 to 9.0 at < 40℃ temperature, and being completely inhibited by the serine protease inhibitor, PMSF, indicated that it belongs to the serine protease family. Specificity test indicated this protease has extensive selectivity to peptide bones, especially to peptide bones composed of Leucine residue.

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

Objective The aim of this study is to investigate the role of angiotensin-converting enzyme (ACE)gene susceptibility to systemic lupus erythematosus(SLE)by familial studies.Methods PCR-based re- striction fragment length polymorphism(RFLP)was applied to genotype single nucleotide polymorphism(SNP) G261T of the ACE gene.A total of 119 patients with SLE from 119 families were recruited.In addition,316 family members of these patients were also genotyped.A family-based association study was carried out to ex- plore the association between gene polymorphism and SLE.We studied the SNP encoding non-synonymous substitution in the ACE gene with respect to genetic susceptibility to SLE.Results Among 119 SLE patients. the frequency of ACEG261TG,T alleles was 44.8%.55.2% respectively,the frequency of ACEG261T GG,GT and TT genotypes was 13.9%,62.0%,24.1% respectively,Univariate(single-marker)family-based association test(FBAT)demonstrated that variant alleles at the SNP,rs4303,exon 5 of ACE gene were significantly asso- ciated with genetic susceptibility to SLE in Additive Model(Z=2.877,P=0.004),Dominant Model(Z=2.557, P=0.011).Recessive Model(Z=2.202,P=0.028).Transmission-disequilibrium test(TDT)and sib transmission -disequilibrium test(STDT)showed an excess of the allele of T from heterozygous parents to affected offspring or higher frequency of the allele of T in the patients than their normal siblings(X~2=11.66,P=0.001).Conclu- sion Our findings suggest that the ACE gene may he the susceptible gene to SLE in Chinese population,and the individuals carrying ACE-261T allele is significantly associated with susceptibility to SLE.

OBJECTIVETo investigate the effect of docosahexaenoic acid (DHA) on invasiveness of aflatoxin B1 (AFB1)-induced hepatocellular carcinoma cells in vitro.

METHODSHepG2.2.15 cells were exposed to different concentrations of AFB1 and DHA plus AFB1. The cell migration and invasion were assessed using wound-healing and Transwell assay, and flow cytometry was used to analyze the cell cycle changes. The ultrastructural changes of the cells were observed by transmission electron microscopy.

RESULTSCompared with the control group, the cells exposed to2 µmol/L AFB1 showed obviously enhanced migration and invasion with decreased cell ratio in G1/G1 phase and increased cell ratio in G2/M phase but no changes in S phase cells; transmission electron microscopy revealed the presence of multiple nucleoli and significantly increased mitochondria and Golgi apparatus in the exposed cells. Compared with AFB1-exposed cells, the cells treated with DHA and AFB1 showed decreased migration and invasion abilities, and the G1/G1 phase cells increased and G2/M phase cells decreased significantly; ultrastructurally, the cells contained single nucleoli with decreased mitochondria and vacuolization occurred in the cytoplasm.

CONCLUSIONDHA can significantly inhibit AFB1-induced enhancement of cell migration and invasion in hepatocellular carcinoma cells in vitro.

Aflatoxin B1 ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Cycle ; Cell Movement ; drug effects ; Docosahexaenoic Acids ; pharmacology ; Golgi Apparatus ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Mitochondria ; Neoplasm Invasiveness

Objective To obtain the elasticity value of solid breast lesions with supersonic shear wave elastrography (SWE) and apply the binary Logistic regression in order to evaluate the value of SWE in differential diagnosis of benign and malignant breast lesions. Methods SWE quantitative elastography was preformed in 91 breast lesions of 91 patients in Zhenghai Longsai Hospital to obtain the maximum and mean elasticity value (Emax, Emean). And receiver operating characteristic (ROC) curves were used to assess the diagnostic performance. A Logistic regression for the gray scale ultrasound and the elastic modulus was conducted with multiple variables including Emax, Emean, border, echo, form, calcification. Results Pathological examination showed 73 benign lesions and 18 malignant lesions. Emax and Emean of malignant lesions were obviously higher than those of benign lesions [(99.73±41.15) kPa vs (38.59±14.28) kPa, (61.45±24.88) kPa vs (23.46±11.44) kPa, t=-15.05,-14.12, both P=0.000]. The area under the ROC curve of Emax and Emean were 0.932 and 0.915. Taking 63.70 kPa as the threshold of Emax, the sensitivity was 77.8%and the speciifcity was 97.3%. Then taking 44.22 kPa as the threshold of Emean, the sensitivity was 83.3%and the speciifcity was 94.5%. The results of Logistic regression analysis showed:the 3 most effective variables were Emax, border of the lesions and Emean. Conclusions The multivariate analysis model of binary Logistic regression can select the valuable indexes of differential diagnosis of benign and malignant breast lesions. SWE plays an important role in differentiating benign and malignant lesions and it is valuable in clinical practice.

Illegally paid blood donation was a risk factor for HIV acquisition exclusively in Henan and Hubei Provinces of China, and not in Shanghai. Nucleotide sequences in the gag and env genes of HIV-1 were compared between isolates from Henan and Shanghai regions of China to test whether an expected higher degree of a common source of infections from this unique blood donation transmission risk would be evident as decreased variation among Henan isolates in an exploratory cross-sectional analysis. Among 38 isolates studied, 23 of 23 (100%) from Henan and 8 of 15 (54%) from Shanghai were subtype B. In addition, fewer sequence differences were found in gp41 of subtype B isolates from Henan than from Shanghai isolates. Further studies with additional controls are therefore warranted to confirm the role of the degree of a common source of infections in differences in HIV variation across populations.

OBJECTIVETo evaluate the effect of alpha-galactosidase A (Gla) deficiency on FV Leiden (FVL) associated thrombosis in vivo.

METHODSTo generate the mice carrying mutations in Gla and FVL and analyze the tissue fibrin deposition in organs and thrombosis.

RESULTSIn the presence of FVL, Gla deficiency greatly increased tissue fibrin deposition compared with that in wild-type [Gla(-/0) FV(Q/Q) vs. Gla(+/0) FV(Q/Q) = (0.24 +/- 0.07)% vs. (0.086 +/- 0.049)%, P < 0.0001; Gla(-/-) FV(Q/Q) vs. Gla(+/+) FV(Q/Q) = (0.32 +/- 0.03)% vs. (0.06 +/- 0.005)%, P < 0.05]. With Gla deficiency, the number of thrombi on organ sections in FVL mice was significantly increased [(Gla(-/-) FV(Q/Q) and Gla(-/0) FV(Q/Q)) vs. (Gla(+/+) FV(Q/Q) and Gla(+/0) FV(Q/Q)) = 1.9 +/- 0.7 vs. 0.3 +/- 0.1, P < 0.05].

CONCLUSIONSGla deficiency could be an important genetic modifier for the enhanced thrombosis associated with FVL.

Animals ; Fabry Disease ; genetics ; Factor V ; genetics ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Mutation ; Thrombosis ; genetics ; pathology

Adolescent ; Arrhythmias, Cardiac ; etiology ; Cardiac Catheterization ; Child ; Child, Preschool ; Electrocardiography ; Female ; Heart Septal Defects, Atrial ; therapy ; Humans ; Infant ; Male