0.05);Single factor analysis showed that,8 factors such as planned pregnancy,gender expectation of mother and father of spouses,relation of primiparas with mother and father of their spouses were independently related to depression in spouse of primiparas;multiple stepwise regression analysis revealed that,social support,books and media with postpartum health care knowledge,gender expectation of mother and father of spouses were main factors that related to stress in spouses of primiparas(?=-0.68,-0.15,-0.13).Conclusion:Spouses of primiparas experience the same medium-leveled stress as primiparas.The related factors were multifold and the main factors were social support,books and media with postpartum health care knowledge,gender expectation of mother and father of spouses.

Objective To analyze and compare the clinical application values of procalcitonin (PCT),leukocyte count (WBC) and C-reactive protein(CRP) in elder patients with infection.Methods In patients(age≥ 65 yrs,axillary temperature ＞38.0℃)with infection or suspected infection,PCT,WBC,CRP and other bacteriological examination were performed.The electronic medical records from the HIS system of our hospital were analyzed retrospectively in combination with medical history.Results Of the enrolled 219 patients,65 ones were in control group,48 ones SIRS,51 ones sepsis and 55 ones MODS.There was a positive correlation between the level of serum PCT and the infection degree.The Spearman correlation coefficient was 0.706 (95％CI:0.616-0.797,P=0.000).Based on the highest Youden index (sensitivity+specificity-1),the best cutoff point of diagnosis for PCT was ＞0.341 μg/L (sensitivity 84.5％,specificity 55.8％),a analysis of receiver operating characteristic(ROC) curve about PCT,WBC and CRP was carried.Area under the curve (AUC) of PCT to controlled infection was 0.916 (95％CI:0.864-0.967,P=0.000).Based on the highest Youden index (sensitivity+ specificity-1),the judging threshold of PCT to infection controlled or not was 0.73 μg/L (sensitivity 84.6％,specificity 88.0％).PCT level after treatment ＞0.73 μg/L showed the uncontrolled infection,＜ 0.73 μg/L controlled.Conclusions PCT has a higher specificity for elder patients with infection.The variation of PCT level can guide the application of antibiotics,avoid abuse and decrease the occurrence of drug-resistant bacteria.

Objective Streptococcus pneumoniae can form biofilms .The aim of this study was to investigate the biofilm forma-tion of Streptococcus pneumoniae and the relationship with antibiotic resistance of penicillin etc . Methods A total of 147 clinical iso-lates of Streptococcus pneumoniae were collected from 7 teaching hospitals in Nanjing from 2010 to 2012.Minimal inhibitory concentration (MIC) of penicillin, erythromycin, cefuroxime and ceftriaxone were determined by agar dilution method .Streptococcus pneumoniae with various penicillin MIC was selected randomly as follow:MIC≤0.065μg/mL, 0.5μg/mL, 2μg/mL and≥4μg/mL, which was incuba-ted to form biofilms in 96-well plates and 24-well plates for 24 hours.The A values at 570 nm was measured and the biofilm was observed through confocal laser scanning microscope ( CLSM) . Results The biofilm semi-quantitative detection and CLSM both displayed that all strains formed biofilms.The A value of the group which penicillin MIC was ≤0.065μg/mL (0.228 ±0.063) was higher than the 0.5μg/mL group (0.200 ±0.061) and the≥4μg/mL group (0.186 ±0.050) (P<0.05) , and there was no difference among the groups which penicillin MIC were 0.5μg/mL, 2μg/mL and≥4μg/mL, respectively (P>0.05).The A value of the group which erythromycin MIC was ≤0.5μg/mL (0.211 ±0.068) was higher than the ≥4μg/mL group (0.201 ±0.052) (P>0.05).The A value of the group sensitive to cefuroxime (0.216 ±0.062) was higher than the group resistant to cefuroxime (0.196 ±0.054) (P<0.05). Conclusion Streptococcus pneumoniae can form biofilms .Streptococcus pneumoniae with high antibiotics MIC has a trend of weakened biofilm formation .

Based on the transcriptome database of Salvia miltiorrhiza, specific primers were designed to clone a full-length cDNA of ent-kaurene oxidase synthase (SmKOL) using the RACE strategy. ORF Finder was used to find the open reading frame of SmKOL cDNA, and ClustalW has been performed to analysis the multiple amino acid sequence alignment. Phylogenetic tree has been constructed using MEGA 5.1. The transcription level of SmKOL from the hairy roots induced by elicitor methyl jasmonate (MeJA) was qualifiedby real-time quantitative PCR. The full length of SmKOL cDNA was of 1 884 bp nucleotides encoding 519 amino acids. The molecular weight of the SmKOL protein was about 58.88 kDa with isoelectric point (pI) of 7.62. Results of real-time quantitative PCR analyses indicated that the level of SmKOL mRNA expression in hairy roots was increased by elicitor oMeJA, and reached maximum in 36 h. The full-length cDNA of SmKOL was cloned from S. miltiorrhiza hairy root, which provides a target gene for further studies of its function, gibberellin biosynthesis and regulation of secondary metabolites.
Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; methods ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Structure, Tertiary ; Salvia miltiorrhiza ; enzymology

To study the correlation between acute lymphoblastic leukemia (ALL) and HLA-A, B and DRB1 gene in southern Chinese Han population and to investigate the susceptible HLA gene to ALL, a total of 4707 healthy volunteer bone marrow donors from southern Chinese Han population were used as a control group, 201 patients diagnosed as patient group from southern Han individuals were genotyped at HLA-A, B and DRB1 loci by PCR-SSP, PCR-SSOP and SBT. HLA allele frequency and its distribution of ALL patient group were compared with the control group by using chi(2) test, and calculated the statistic value of relative risk (RR), pathogenicity score (EF) and preventive score (PF). The results showed that in comparison with the control group, the gene frequence of HLA-A26, B56 and DR9 increased significantly, but the gene frequence of HLA-A30, A33 and B58 allele frequency decreased significantly for patients with ALL. It is concluded that HLA-A26, B56 and DR9 gene have a high correlation with ALL and seem to contribute the genetic susceptibility to ALL in southern Chinese Han populations. However, HLA-A30, A33 and B58 gene seem to have protective role for southern Han individuals suffered from ALL.
Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Chi-Square Distribution ; Child ; Child, Preschool ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; ethnology ; genetics

Objective To investigate the association of single nucleotide polymorphisms (SNPs) in the SCGB3A2(secretoglobin family 3A member 2) gene promoter with susceptibility of Graves' disease.Methods One-hundred and seventy-nine SNPs within a 3.0 Mb region surrounding marker D5s2090 were scanned in a case-control study.The size of the region(s) associated with GD was then narrowed.Results Total 179 SNPs within a 3.0 Mb region surrounding marker D5s2090 were analyzed.The most significant association signal was found at SNP rs1368408 (P =3.69 × 10-5).Subsequent association analysis was then performed and the results suggested that the SNP76 (P =4.11 × 10-8) and SNP75 (P =1.37 × 10-8) in the promoter of SCGB3A2 gene may be the causal variants of GD.Logistic regression analysis suggested these 2 SNPs in this region may contribute to GD susceptibility.Conclusion A significant association seems to exist between GD with the SCGB3A2 gene.

OBJECTIVETo establish a reliable assay for cloning and sequencing the full-length HLA-Cw gene.

METHODSIn this study, a fragment of 4.5 kb full-length HLA-Cw gene was amplified using the self-designed PCR primer pair by long template PCR, purified PCR products was cloned into the pGEM-Teasy plasmid vector and the plasmid DNA isolated from positive clones was subjected to haplotype sequencing by both directions. A total of 12 samples having been previously-genotyped by PCR sequence-based-typing (PCR-SBT) were amplified by using the TaKaRa LA Taq and Stratagene Pfu polymerase, respectively. PCR products of full length HLA-Cw gene were subjected to cloning and sequencing and the obtained haplotype sequence were compared with the PCR-SBT results.

RESULTSThe specific target fragment of HLA-Cw gene could be amplified and the full-length HLA-Cw allele sequence covering from nucleotide position -962 in 5'untranslated region (5'-UTR) to nucleotide position 3576 in downstream area of 3'-UTR region could be obtained using our method. The results of cloning and sequencing analysis indicated that the Stratagene Pfu polymerase had better fidelity than the TaKaRa LA Taq polymerase in this experiment. By comparing the sequences of Cw*07020101 with Cw*010201, 11 SNPs as well as 2 insertions/deletions in nt-962--284 of 5'-UTR, and 11 SNPs as well as 1 insertion/deletion in nt3067-3576 downstream of 3'-UTR were identified.

CONCLUSIONOur results indicate that the technique for cloning and sequencing full-length HLA-Cw gene has been established, it has a broad application in full-length HLA-Cw gene polymorphism study and the regulation and expression of HLA-Cw gene.

Alleles ; Amino Acid Sequence ; Base Sequence ; China ; ethnology ; Cloning, Molecular ; methods ; DNA Primers ; HLA Antigens ; genetics ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA ; methods

Thirst study was purposed to explore the genetic polymorphism of Chinese Zhuang population at HLA-Cw locus by sequence based typing (SBT). A total of 150 unrelated blood samples from Chinese Zhuang population were subjected to sequencing at exon 2, 3 and 4 of HLA-Cw gene in both directions by using SBT technique established by our laboratory. The purified products of sequencing reaction were run by means of electrophoresis on the ABI 3730 DNA Sequencer and the assignment of HLA-Cw genotype was accomplished by using the Assign 3.5 software. The consensus sequence at exon 2, 3 and 4 of HLA-Cw gene for each sample was imported into the Assign 3.5 software. The results showed that 33.33% of tested samples could obtain an unique genotype, genotype in 63.33% of tested samples with ambiguous results could be assigned by ruling out the rare alleles according to the NMDP Rare Allele List File; however, the final genotype in rest 3.33% of the detected samples could be defined when subjected to further confirmatory testing by PCR-SSP. In this detection 16 HLA-Cw alleles were identified, the common alleles with a frequency of > 10% were Cw*0304 > Cw*0102 > Cw*0801 > Cw*0702. The value for gene diversity (GD) was 0.9297, The frequency for Cw*01, 03, 07, 08, 12, 14 (Cw 1 allele group) and Cw*02, 04, 05, 06, 15, 16, 17, 18 (Cw 2 allele group) was 0.8967 and 0.1032, respectively, which indicated that the Cw 1 allele group is the dominant ligand for KIR in Chinese Zhuang population. 51 genotypes were determined and the distribution of genotype frequency was in line with Hardy-Weinberg principle. It is concluded that the obtained HLA-Cw allele frequency and its distribution characteristics of Chinese Zhuang population can provide valuable data in the studies of anthropology and the association of HLA-Cw with disease.
Asian Continental Ancestry Group ; genetics ; Exons ; Gene Frequency ; Genotype ; HLA-C Antigens ; genetics ; Humans ; Molecular Sequence Data ; Polymorphism, Genetic ; Sequence Analysis, DNA

OBJECTIVETo identify a novel human leukocyte antigen (HLA) allele by cloning and sequence-based typing in Chinese population, and analyzing the sequence of the introns 1 and 2.

METHODSThe routine HLA-A, -B, -DRB1 low resolution genotyping for stem cell donor from Guangdong province was performed with polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP). An unknown HLA-DRB1 allele was initially detected by HLA typing. Genomic DNA of the proband was amplified by using HLA-DRB1 locus group-specific primer, the amplified product was cloned, sequenced, and compared to the closest DRB1*120201 allele and the closest intron sequence of the DRB1*030101 allele.

RESULTSThe sequencing results showed that a normal DRB1*080302 and a novel DRB1*1218 variant allele were identified. The sequence of the novel allele has been submitted to GenBank (FJ481086). The novel allele had 1 nucleotide substitution of the closest matching allele HLA-DRB1*120201 at nt262(G-->C) in exon 2,resulting in an amino acid change from Glu(GAG)-->Gln (CAG) at codon 59.The intron 2 sequence is identical between the novel HLA-DRB1*1218 and DRB1*030101, but there are 12 nucleotides substitution in intron 1.

CONCLUSIONA novel HLA allele was confirmed by cloning and sequence-based typing in Chinese. It was officially designated as HLA-DRB1*1218 by WHO Nomenclature Committee.

Alleles ; Amino Acid Substitution ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Cloning, Molecular ; Exons ; genetics ; Glutamic Acid ; genetics ; Glutamine ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Molecular Sequence Data

OBJECTIVETo analyze the polymorphism and haplotypes of HLA-A, B, Cw, DRB1 and DQB1 loci in Chinese Han population.

METHODSA total of 186 unrelated healthy individuals from southern China were analyzed by sequence-based typing. Two-, three-, and five-locus haplotypes were estimated using the Expectation Maximization Algorithm. RESULTST: Twenty-eight alleles for the HLA-A locus, 49 HLA-B alleles, 24 HLA-C alleles, 29 HLA-DRB1 alleles and 20 HLA-DQB1 alleles were detected. The A*0207-B*4601(10.81%), A*3303-B*5801(6.14%), B*4601-DRB1*0901(6.22%), B*4001*-DRB1*0901(3.78%), DRB1*090-DQB1*0303 (12.16%) and DRB1*1202-DQB1*0301(8.38%), A*0207-B*4601-Cw*0102 (10.75%), A*3303-B*5801-Cw*0302 (5.14%), A*0207-B*4601-DR*0901(5.07%), A*3303-B*5801-DRB1*0301(2.96%), A*0207-B*4601-Cw*0102-DRB1*0901-DQB1*0303(4.87%) and A*1101-B*1301-Cw*0304-DRB1*1501-DQB1*0601(2.43%) were the most common haplotypes in the southern Chinese Han population.

CONCLUSIONThe results have shown the characteristics of the five HLA loci haplotype distribution and provided more information in anthropology, disease association studies and transplantation.

Adult ; Alleles ; Base Sequence ; China ; ethnology ; Female ; Genetics, Population ; HLA Antigens ; analysis ; genetics ; immunology ; HLA-B Antigens ; analysis ; genetics ; HLA-DQ Antigens ; analysis ; genetics ; HLA-DQ beta-Chains ; HLA-DR Antigens ; analysis ; genetics ; HLA-DRB1 Chains ; Haplotypes ; Humans ; Male ; Population Groups