Objective To establish a method for detection of RhD(+) red blood cells mixed in D-negative blood by flow cytometry(FCM).Method RhD(+) and RhD(-) RBCs were mixed according to predefined ratios.Cells were indirectly labeled,with IgG anti-D labeled as the first antibody,and FITC-anti-IgG F(ab')2 as the second antibody.The percentage of RhD(+) RBCs was determined by FCM,and the best dosage of IgG anti-D was also defined.The ratio of red cells in the two groups,measured by FCM,was compared with the actual ratio.The consistency of method was also evaluated.Results The effective dosage of IgG anti-D was 1∶4,and 50?l/1?106 cells.When the actual percentages of RhD(+) cell among RhD(-) cells were 2.5%-0.312%,the correlation coefficient between the percentages measured by FCM and the actual percentages was 0.987.The same tubes,containing 10% and 2.5% RhD(+) RBCs,were each tested for 10 times,and their coefficient of variation were 3.4%,and 4.9%,respectively.Conclusion The method of quantifying the RhD(+) RBCs in D-negative blood by FCM is feasible and repeatable,which deserves a further clinical application.

Enterovirus type 71(EV71) causes severe hand-foot-and-mouth disease(HFMD) resulting in hundreds of deaths of children every year; However,currently,there is no effective treatment for EV71.In this study,the EV71 poly-protein(EV71-P1 protein) gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GS115.The EV71 P1 pretein with a molecular weight of 100 kD was produced and secreted into the medium.The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70％.The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits.We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.

Objective To investigate the relationship of matrix metalloproteinases(MMP?3 and MMP?9)and the tissue inhibitor TIMP?1 with left ventricular hypertrophy(LVH)in patients with primary hypertension. Methods Totally 140 patients with primary hypertension and 132 healthy controls were included. Matrix metalloproteinase?3(MMP?3),matrix metalloproteinase?9(MMP?9)and tissue inhibitor metalloproteinase?1 (TIMP?1)were measured. All subjects were taken echocardiography examination ,then left ventricular mass index(LVMI)was calculated. Re?sults MMP?3,MMP?9,TIMP?1(488.32±100.32 vs 314.59±99.78;340.56±43.21 vs 290.15±33.98;389.16±57.53 vs 243.45±62.31;P<0.001) and LVMI(113.7±9.9 vs 88.3±10.4,P<0.001)in patients with primary hypertension were significantly higher than those in controls. In a multiple stepwise regression analysis with LVMI as the variable,it was found that age,SBP,MMP?3,MMP?9 and TIMP?1 were main determinants for LVMI (r2=0.78,P<0.001). 3. Patients with primary hypertension were divided into two subgroups according to LVMI,i.e.,hypertension with LVH (group A)and hypertension without LVH(group B). SBP,MMP?3,MMP?9 and TIMP?1(178±31 vs 166±25;490.14±99.13 vs 405.56±53.12;340.56±43.21 vs 290.15±33.98;393.45±47.69 vs 301.58±39.57;P<0.05)of group A were significantly higher than those of group B. Conclusion MMP?3,MMP?9 and TIMP?1 are influencing factors for LVH in patients with primary hypertension.

Objective To investigate the diagnostic value of breast MRI in patients presenting with microcalcifications on mammography.Methods Eight four patients were retrospectively analyzed,who had mammographically detected BI-RADS (breast imaging reporting and data system) 3 to 5 microcalcifications and underwent breast MRI before surgical biopsy.All mammography and MR images were reviewed with BI-RADS.With histopathological diagnosis as golden standard,the sensitivity,specificity and accuracy of the two methods were calculated and compared with x2 test or Fisher exact test.The diagnostic efficacy of the two methods was compared with ROC curve.Results Pathologic examination revealed 91 lesions in 84 patients including 49 benign lesions and 42 malignant lesions.For 21 lesions of category 3 microcalcifications,the specificity of mammography and MR was 100.0％ (21/21) and 95.2％ (20/21),which had no significant difference (P=1.000).For 51 lesious of category 4,sensitivity,specificity and accuracy of mammography were 100.0％(23/23),0 and 45.1％(23/51).The corresponding values for MR were 91.3％(21/23),82.1％ (23/28) and 86.3％ (44/51).The difference for specificity and accuracy between the two methods was statistical significant(x2 value was 30.030 and 19.182,respectively,with P＜0.01),but not for sensitivity(x2=0.523,P=0.470).Nineteen lesions of category 5 were all correctly diagnosed on mammography and MRI.For all the 91 lesions,the sensitivity,specificity and accuracy of mammography were 100.0％(42/42),42.9％(21/49) and 69.2％(63/91),respectively.The corresponding values for MRI were 95.2 ％(40/42),87.8％(43/49) and 91.2％(83/91).There was significant difference for specificity and accuracy between the two methods (x2 value was 21.798 and 13.851,respectively,with P＜0.05),but not for sensitivity (x2=0.512,P=0.474).The areas under ROC curve for mammography and MR were 0.844,0.945(P＜0.01),for the estimation of the benign and the malignent.Conclusions Compared with mammography,breast MRI significantly improved the diagnosis of category 4 microcalcifications with increased specificity and accuracy.But for microcalcifications of category 3 and 5,MR didn't improve the diagnostic effect.

AlM: To research the characteristics of positive relative accommodation ( PRA) , negative relative accommodation (NRA) and PRA/NRA ratio in myopes. To analyze the relationship among PRA, NRA, PRA/NRA ratio, spherical equivalent degree, years and habbits of wearing glasses, myopia development, and pupil diameter.METHODS: Aretrospective study of ninety eyes in the 180 th Hospital of Quanzhou from August 2014 to December 2014. PRA, NRA and PRA/NRA ratio were compared among low, moderate, high myopes and emmetropes. The correlation were analyzed among PRA, NRA, PRA/NRA ratio, spherical equivalent degree, years and habbits of wearing glasses, myopia development and pupil diameter. PRA, NRA, PRA/NRA ratio, years and habbits of wearing glasses and pupil diameter were compared between progress group and non-progress group.RESULTS: ( 1 ) Without statistical differences in age, sex and intraocular pressure, PRA and PRA/NRA ratio of myopes were lower than emmetropes, while NRA was higher. (2) Without statistical differences in age, sex and intraocular pressure, PRA, PRA/NRA ratio and NRA had no statistical differences while years and habbits of wearing glasses had statistical differences among low, moderate, high myopes. ( 3 ) With longer years of wearing glasses, PRA, PRA/NRA ratio were larger and NRA, pupils were smaller. ( 4 ) Without statistical differences in age, diopter and intraocular pressure, one group which were not easy to deepen degree had more often-wear-glasses myopia patiens and longer years of wearing glasses, the other group which were easy to deepen degree had more seldom-wear-glasses myopia patiens and shorter years of wearing glasses.CONCLUSlON: PRA and PRA/NRA ratio of myopes were lower than emmetropes, while NRA was higher. No correlated relation was detected among PRA, NRA, PRA/NRA ratio, spherical equivalent degree and myopia development. lt suggests the onset and progress of myopia are related to many factors. Wearing-glass timely and accurately can release the decline of PRA and PRA/NRA ratio and slow down degree development in myopes.

Objective To investigate the expression of steroidogenic factor 1 ( SF-1 ) in eutopic endometrium,ovarian endometriosis (EM) and adenomyosis.Methods From Jan.2008 to Dec.2010,30 patients with both ovarian endometriotic cysts and adenomyosis underwent hysterectomy,cystectomy or adenxectomy in Peking University First Hospital.The combination of ovarian EM and denomyosis were confirmed in 17 cases.In the mean time,endometria form 10 patients with cervical intraepithelial neoplasm ( CIN ) Ⅲ underwent hysterectomy were selected as controls.The expression of SF-l in eutopic endometrium,ovarian EM and adenomyosis was detected by immunohistochemistry staining.Results No immunoreactivity of SF-l was observed glandular cell and stromal cell in eutopic endometrium of both groups.14/17 of positive rate of SF-l were observed in lesions of ovarian endometriosis.SF-l protein was not expressed in the endometrial glandular cells of ovarian EM and in adenomyosis foci.Conclusion The differences of SF-l expression in ovarian endometrioma and adenomyosis foci might mediate the development of the disease.

Objective: To examine the expression of glucocorticoid receptor in human colon cancinoma tis-sues and call lines and to explore the survival of colon cancer cell lines treated with dexamethasone alone or in combination with 5-Fu and L-OHP in a way of dexamethasone pretreatment or co-administration in vitro.Methods: The expression of glucocorticoid receptor was detected in 61 cases of colon cancer tissue samples and 4 types of colon cancer cell lines by immunohistochemistry. Apoptosis was detected by Hoechst33342 staining and flow cytometry. MTT assay was employed to detect the chemosensitivity of colon carcinoma cells to L-OHP and 5-Fu with dexamethasone pretreatment for 24 hours or co-administretion. Results: Positive GR expression was found in 57.3% colon cancer tissue samples and in Lovo and HCT-116 cell lines, not in HT-29 and SW-480. Apoptosis was detected in GR-expressed Lovo and HCT-116 cells at 72 hours after 1×10~(-4)mol/L Dex treatment, and the rates of apoptosis were higher than those in the control groups without Dex (P<0.01),GR-negative cells, HT-29 and SW-480 even treated with 1 × 10~(-4)mol/L Dex for 72 hours. Pretreatment and co-administration for Lovo cells with 1×10~(-4)mol/L Dex could decrease the IC50 of L-OHP from 13.7±1:1.3μg/mL to 5.9±0.6μg/mL and 4.8±0.7μg/mL, respectively. IC50 of 5-Fu was decreased from 72.2±8.1 μg/mL to 21.1±4.1μg/mL and 18.6±4.0μg/mL, respectively. Conclusion: There is expression of glucocorticoid receptor in part of colon carcinoma tissue samples and cell lines. Apoptosis does occur in GR-expressed Lovo and HCT-116 cells induced by dexamethasone in vitro. Pretreatment for 24h and co-administration with Dex can increase the chemosensitivity of Lovo cells to L-OHP and 5-Fu.

In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae ; classification ; genetics ; Base Sequence ; China ; DNA, Intergenic ; genetics ; Gene Flow ; Genetic Variation ; Molecular Sequence Data ; Phylogeny

With the successful implementation of Human Genome Project, more and more scientists started to pay attention on the second genome of human body: Microbiome. This paper will briefly introduce the latest developments of the Human Microbiome Project, the human oral microbiome research, and new technologies of microbial gene research.
Humans ; Metagenome ; Microbiota ; Mouth ; microbiology

Objective To investigate the prevalence of plasmid-mediated quinolone resistance ( PMQR ) determinants [ qnr,aac ( 6' ) -Ib-cr and qepA ]and mutations in quinolone resistance-determining regions (QRDRs) of gyrA and parC and their association with fluoroquinolone susceptibility in clinical isolates of Serratia marcescens in Anhui.Methods The minimum inhibition concentration ( MIC ) of 104 strains of S.rnarcescens collected from various clinical specimens from 34 hospitals during 2005 to 2010 were determined by agar dilution method.The qnr,aac (6')-Ib,qepA,gyrA and parC genes were screened by polymerase chain reaction (PCR) in 31 strains resistant to ciprofloxacin,and positive results were subsequently confirmed by sequencing.The conjugation experiments were performed for qnr and aac(6')-Ib-cr positive strains.The MIC of S.marcescens isolates,recipient strains and conjugants were tested by agar dilution method for quinolones and other antimicrobial agents.Results Six strains of the 31 S.marcescens isolates harboured qnr and/or aac(6')-Ib-cr genes.Among those 6 strains,2 strains harboured a qnrB6 gene,1 harboured a qnrS2 gene,and 4 harboured aac( 6' ) -Ib-cr,whereas no qnrA-,qnrC- or qnrD-positive isolate was detected.None of the 31 isolates carried the qepA gene.Mutations in the QRDR of gyrA and parC genes were detected in 9 and 7 isolates,respectively.The conjugation experiments were successfully carried out in 5 isolates of 6 PMQR determinants-postive strains.The MIC of conjugants for quinolones were increased evidently compared to recipient strains.Conclusions Chromosome and plasmid-mediated resistance determinants play an important role in quinolone resistance in clinical isolates of S.marcescens.And more important is that the PMQR determinants can be horizontal transmitted.It is necessary to continuously survey and watch for the spread of PMQR in S.marcescens in public health control program.