Objective To investigate the immunomodulatory effect of black rice anthocyanin(BA) on biochemical indicators in blood and macrophages in rats.Method Twenty four female SD rats were randomly divided into 4 groups:control group,BA low,middle and high-dose groups(BAL 25;BAM 50;BAH 100 mg/kg bw).The control group was treated with normal saline,while BA groups were i.g.administered with defferent doses for 30 d.The blood biochemical indicators were detected by automatic biochemistry analyzer.The activities of lactate dehydrogenase(LDH),acid phosphatase(ACP),superoxide dismutase(SOD),and maleic dialdehyde(MDA) contents in peritoneal macrophage(PM) and alveolar macrophage(AM) were determined by biochemical methods.The ability of macrophages to phagocytose neutral red was measured by colorimetric method.Results The levels of urea nitrogen(UN),uric acid(UA),triglyceride(TG),total cholesterol(TC) and low density lipoprotein(LDL) in blood were reduced.The activities of ACP,LDH in PM were increased.and the activities of SOD in PM and AM were increased whereas MDA was reduced.The ability of PM and AM to phagocytose neutral red was strengthened significantly.Conclusion BA shows significant immunomodulatory effection blood biochemical indicators and macrophages in rats.

Objective To establish a method for high throughput screening single nucleotide polymorphism(SNP) by tag microarray,and then apply the method to study the gene SNP which is related to the motor function of normal people.Methods The genes related to motor function were firstly defined,and then 48 SNP loci were determined.The rs numbers of these SNP loci were fingered out from PubMed,and the primers were designed with the software in web site "www.autoprimer.com".The primer sequences were then downloaded and sent to the biologic corporation for synthesis.After being synthesized and purified by HPLC these primers were used in the experiments according to the instruction of Bakeman's SNPstream machine.The key techniques of SNPstream machine were tag microarray and single nucleotide extension assay.Once the determination was finished,both the gene frequency and allele frequency of every locus could be statistically analyzed.Results The information of the 48 SNP loci that related to motor function had been determined simultaneously by tag microarray,regardless the number of samples to be detected at the same time.The number of the samples was variable to meet the need.The data of gene frequency and allele frequency of these 48 SNP loci may be used in the subsequent studies.Conclusions Tag microarray used to high throughput screening SNP has the advantages of accuracy,speed,efficiency and reasonable cost.Therefore it can be applied to study the relationship between the SNP and many kinds of diseases.

A method was developed for the quantification of human growth hormone ( hGH ) by protein purification and isotope dilution-high performance liquid chromatography mass spectrometry ( HPLC-IDMS ) . The hGH was purified and fractionated by fast protein liquid chromatography ( FPLC ) , then hGH molecular weight was accurately determined by Fourier transform ion cyclotron resonance mass spectrometer ( FTICR-MS). The purified hGH was hydrolyzed and the separation was performed on an KINETEX C18 column (150 mmí2 mm I. D. , 2. 6 μm) with water ( containing 0. 1% TFA) and acetonitrile isocratic solution as the mobile phase at a flow rate of 0 . 2 mL/min and 40℃. The electrospray source was operated in the positive ion mode, and monitored in the multiple reaction monitoring ( MRM) mode. The measured hGH molecular weight by FTICR-MS was only 0. 31 Da difference from theoretical value. Three amino ( proline, valine and phenylalanine) were clearly separated by isocratic elution within 5 min. Under the optimized conditions, the content of hGH was 186 . 80 μg/g with a RSD of 0 . 5%. The detection results of hGH in international comparison by this method were consistent with the reference value, which validated the feasibility of the established method. The developed method is simple, practical, accurate, reliable and reproducible, and can be used for the hGH quantitation of pure hGH CRM to provide reference for the routine detection of hGH.

Objective To assess the CT image quality of the pure ground glass nodule(pGGN)in chest phantom by using different field of view(FOV)and matrix.Methods CT(Philips Brilliance 128 spiral CT)scans were performed on chest phantom containing 9 artificial pGGNs(diameter≥5 mm)in 3 different FOV (50 mm,150 mm,300 mm ),and were reconstructed in 2 different matrix(512×512, 1 024×1 024),standard kernel.Recorded the CT values and standard deviations (SD)of the nodules and surrounding regions in different FOV and matrix,and calculated the mean standard deviation(MSD),contrast noise ratio (CNR)and signal noise ratio (SNR),then compared the difference among them.Two radiologists assessed the image quality of the pGGNs in blind method respectively,then evaluated the coherence between them using Kappa testing.Results The coherence of 2 observers was substantial or almost perfect.No significant differences were found on MSD,CNR and SNR in different FOV (P value>0.05)when matrix was kept,whereas the visibility of the nodules improved with the FOV changing smaller.And there were significant differences all on MSD,CNR and SNR by using different matrix when FOV was kept.The MSD increased and the CNR,SNR decreased in high-resolution group.The visibility of the pGGNs was not improved obviously in high-resolution group.Conclusion The image quality of the pGGN(diameter≥5 mm)won’t be changed by using smaller FOV when matrix is kept ,but the visibility will be improved with the FOV changing smaller.When FOV is kept,the MSD increased and the CNR,SNR reduced in high-resolution group,but the visibility will not be improved obviously compare to the standard resolution group.

Objective:To study the inhibitory effect of retinoblastoma 94(Rb94) gene combined with radiotherapy ionizing radiation on the growth of esophageal carcinoma cells of tumor-bearing nude mice, and to clarify the synergistic effect of Rb94 gene and radiotherapy in inhibiting the growth of tumor cells.Methods:The models of tumor-bearing BALB/c-nu nude mice were built by inoculating the K150 cells.The model mice were divided into five groups:blank control(no any treatment), Ad-LacZ(control adenovirus including LacZ gene but not Rb94 gene, Ad-LacZ was transfered into tumor xenograft on 0, 3, 7 d separately), Ad-Rb94(tumor xenograft was transfected with Ad-Rb94 on 0, 3, 7 d separately), radiation (tumor xenograft was irradiated with 4 Gy γ-radiation on 1, 4, 8 d separately) and Ad-Rb94 combined with radiation(combination group, tumor xenograft was irradiated with 4 Gy γ-radiation after transfected with Ad-Rb94) groups.The volumes and the weights of esophageal carcinoma and the inhibitory rates of tumor growth of the mice in various groups were detected.The expression levels of ABL and JNK kinase in tumor tissue of the mice in various groups were measured,and the pathological changes of tumor tissue were investigated.Results:The speeds of tumor growth of the nude mice in Ad-RB94, radiation, and combination groups were slower than that in control group.The volume of esophageal carcinoma in combination group at day 15 after treatment was markedly smaller than those in Ad-RB94 and radiation groups,and there were significant differences compared with control group and Ad-LacZ group (F=26.7,23.8;P<0.01).The tumor weight of the nude mice in combination group was the lightest at the end of treatment;the inhibitory rate of tumor growth in combination group reached 81.16% and was significantly higher than those in Ad-Rb94 group(57.84%)and radiation group(38.20%)(P<0.01).The expression levels of ALB and JNK kinase in tumor tissue of the mice in combination group was markedly higher than those in control group(P<0.01).Compared with other groups, the tumor cells in combination group had fewer karyokinesis and lower level of nuclei hyperchromasia.Conclusion:Rb94 gene combined with radiotherapy shows synergistic effect in inhibiting the growth of tumor of tumor-bearing nude mice.

Objective To inVestigate the Potential mechanism and ProtectiVe effects of osthole on rats with focal cerebral ischemia_rePerfusion injury. Methods Middle cerebral artery occlusion was induced by the suture_occluded method. After 2 h of cerebral ischemia and 24 h of rePerfusion,scores of neurological deficits and infarct Volume were eValuated. The leVels of SOD,MDA,GSH and ATPase were also determined. Results Scores of neurological deficits and infarct Volume were lower in the osthole grouPs than in the model grouP. Meanwhile,the actiVity of SOD and GSH content were increased,while the MDA content was decreased in osthole grouPs. Conclusion Osthole exerts ProtectiVe effects on rats with focal cerebral ischemia_rePerfusion injury.

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Background Laser in situ keratomileusis (LASIK) is frequently performed to reduce or eliminate myopic refractive error.Some patients complain of a loss of visual function after surgery even when they have 20/20 visual acuity.One of the reasons is the change of asphericity of the cornea.Objective This study attempts to investigate the short-term changes of the anterior and posterior corneal asphericity after LASIK.Methods One hundred and seven myopic eyes of 54 subjects with the equivalent spherical diopter of -1.30 to -7.50 D who have received LASIK were enrolled in this prospective study.The Q-values of the posterior corneal surface for different corneal diameters (6mm,7mm,8mm and 9mm) were measured with Pentacam preoperatively and 1 month postoperatively.The correlations between Q-value,Q change (△Q),and the mean preoperative spherical equivalent refraction (SE),central corneal thickness (CCT),central ablation depth (AD) and residual bed thickness were investigated.Written informed consents were obtained from all the subjects prior to the clinical trial.Results The Q-values of the anterior and posterior corneal surfaces gradually decreased to negative values with the increase of corneal diameter in myopic eyes.Weak correlations were found between the asphericity of the anterior and posterior corneal surfaces from diameters of 6mm,7mm,8mm and 9mm (r=0.227,0.288,0.303,0.389;P=0.019,0.003,0.002 and 0.000,respectively).No statistically significant correlation was found between Q-value and the diopter of refractive error (P＞0.05).Both the anterior and posterior corneal Q-values varied toward the positive direction except that in the 9mm area of the posterior corneal surface at postoperative 1 month(t=-1.495,P=0.138).The increase of the anterior corneal asphericity (△Q) was more obvious than that of the posterior corneal surface and showed a positive correlation with ablation depth and a negative correlation with residual bed thickness.However,no statistically significant correlation was seen between △Q and these two parameters in the posterior corneal surface (P＞0.05).Conclusion The shape of the anterior and posterior corneal surface shows more prolateness as the increase of corneal diameter in myopic eyes.Both the anterior and the posterior corneal surfaces have an oblate shift within the ablation zone 1 month after LASIK.

Objective To detectthe phenotype and gene mutations underlying aninherited dysplasminogenemia pedigree and search the virulence gene.Methods The peripheral venous blood samples of the proband and his family members (fourteen subjects of three generations in total) were collected,and their prothrombin time(PT),activated partial thromboplastin time(APTF),thrombin time(TT),fibrinogen (FIB),fibrinogen degradation products (FDP),D-dimmer (D-D)weretested on a STAGO analyzer,the plasminogen activity (PLG:A) and plasminogen antigen (PLG:Ag) were analyzedby thechromogenic substrate assay and rocket immunoelectrophoresis,respectively.All 19 exons,5' and 3' untranslated regions of PLGwere amplified with PCR.Direct DNA sequencing was used to analyze the amplified products,which wereconfirmed by backward sequencing.Three bioinformatics online softwares (SIFT,PolyPhen-2 andMutationTaster) were used to forecast the possible impact of the mutations on the protein function.At last,themodel analysis of mutate site was taken on a Swiss-Pdb Viewer software.Results The PLG:Avalue of theproband and other 6 family members were decreased to the half,while the PLG:Ag was normal.The D-Dand FDP value of the proband,his grandma and father were slightly higher.DNA sequencing has revealedthat the proband and the other 6 members of this family had the same mutation of g.38829G ＞ A in exon 15,leading to the missense mutationp.Ala601Thr.The results of bioinformatics softwares showed that themutation could affect the thePLGfunction.Protein model analysis indicated that the hydrophobic interaction force and hydrogen bond between the amino acids were changed,which might affect the stability of the PLG.In addition,all the members of this family take the heterozygous SNP of g.2501C ＞ A in the 5 ＇UTR.Conclusions The p.Ala601Thr found in the inherited dysplasminogenemia pedigree in the exon 15 was responsible for the reduced PLG:A of the family,the dysplasminogenemia and this mutation were both reported for the first time in China.

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .