Objective:To evaluate the immunogenicity of the recombinant plasmid pcDNA 3.1(+)/kgp_ cd.Methods:BALB/c mice were immunized with recombinant plasmid pcDNA 3.1(+)/kgp_ cd,KGP_ cd protein(control) or vector pcDNA 3.1(+)(control) by quadriceps injection or targeted submandibular gland(TSG) injection. Serum IgG and salivary sIgA levels were assayed by indirect ELISA after immunization. The expression of the protein KGP_ cd in quadriceps and submandibular gland was detected by immunohistochemistry techniques.Results: Serum specific anti-KGP_ cd IgG elicited by pcDNA 3.1(+)/kgp_ cd and KGP_ cd protein was significantly higher in both the quadriceps injection group and the TSG group than that in the pcDNA 3.1(+) group (P

The effects of polymerization conditions including scan potential range, scan cycles, the concentration ratio of template molecules to functional monomer, pH of the buffer, and washing time for removing the template molecule from the imprinted polymer on the difference of zero current potential of benzidine ( BZ) interaction with BZ-MIP were investigated. The optimum preparations were obtained. The imprinted capacity of benzidine, 4-chloroaniline, and 4-aminobiphenyl and carmine was calculated as 0. 632, 0. 1123, 0. 1123, 0. 0847 and 0. 0725, respectively. This indicated that BZ-MIP had good specific recognition and selectivity to benzidine, and other substances did not interfere with the binding of BZ-MIP with BZ. The zero current potential variation was linear with the lorgarithm of BZ concentration in the range of 4í10-8-1í10-5 mol/Lwith detection limitation of 1. 89í10-8 mol/L. The sensor was used to detect BZ in waste water sample with recoveries of 95 . 7%-104 . 2%.

OBJECTIVETo study the effect of 905A to G mutation of α -1,3 galactosyltransferase of ABO gene on B antigen expression.

METHODSThree samples were diagnosed as B subgroup by serological test. Genotyping and sequencing were performed with polymerase chain reaction-sequence specific primer (PCR-SSP), direct sequencing and gene dones of exons 6 and 7 of the ABO locus.

RESULTSThe sequence of B allele has differed from that of regular B101 allele with a 905A to G missense mutation in exon 7, which resulted in an amino acid substitution (D302G) in all of the three B subgroup samples.

CONCLUSION905A to G mutation can reduce the expression of B antigen.

ABO Blood-Group System ; genetics ; Alleles ; Amino Acid Substitution ; Base Sequence ; Female ; Galactosyltransferases ; genetics ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense

Objective To investigate the changes of fatty acid oxidase in the placenta of preeclampsia cases with different clinical features, and the relationship with oxidative stress and inflammatory response. To study the correlation of serum free fatty acid (FFA) and triglycerides (TG) level in early second trimester with the molecular changes of the long-chain fatty acid oxidase in the third trimester. Methods This was prospective cohort study, in which cases with singleton pregnancies who archived in Haidian Maternal and Children′s Hospital, Beijing, from January 1st 2012 to May 31st, with regular prenatal care were included. Doppler ultrasound was used for screening for the presence of early diastolic notch of uterine artery at 22-24 weeks of gestation. All the 101 cases with the early diastolic notch of uterine artery were included as the notch group, and 377 cases without the early diastolic notch of uterine artery were included as the non-notch group. The perinatal outcomes and the incidence of hypertensive disorders in pregnancy of the two groups were observed. The serum level of FFA and TG was tested, and the mRNA and protein expression of long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD), P47-phox subunit of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, p38 mitogen-activated protein kinase α (p38MAPK-α) and cyclooxygenase-2 (COX-2) were detected using real-time quantitative PCR and western blot. The relationship between serum level of FFA and TG and the mRNA and protein expression of LCHAD, NADPH P47-phox,p38MAPK-α and COX-2 of the placental tissue specimens were analyzed. Results (1) In the notch group, there were 9 cases of early-onset preeclampsia,15 cases of late-onset preeclampsia and 10 cases of gestational hypertension;and there were 8 cases of late-onset preeclampsia and 18 cases of gestational hypertension in the non-notch group. 15 cases with normal blood pressure in each group were randomly selected as the control group.(2)The serum level of TG of cases of early-onset preeclampsia, late-onset preeclampsia and gestational hypertension in the notch group were(2.0±0.8),(1.8±0.6)and (1.9±0.7)mmol/L, and that of FFA were(0.68±0.26),(0.52±0.10)and(0.52±0.17)mmol/L, respectively. The serum level of TG of cases of late-onset preeclampsia and gestational hypertension in the non-notch group were(1.6±0.6)and(1.4±0.4)mmol/L, and that of FFA were(0.49±0.11)and(0.48±0.05)mmol/L, respectively. The serum level of TG and FFA in the control group were(1.4±0.5)and(0.52±0.06)mmol/L, respectively. The TG level of the notch group was higher than that of the control group, and the difference was statistically significant (P<0.05). The FFA level of the early-onset preeclampsia in the notch group was higher than that of late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and the control group, and the difference was statistically significant (P<0.05).(3) The mRNA expression of LCHAD in the placenta of early-onset preeclampsia in the notch group was significantly lower than that of the late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and the control group (P<0.01). The mRNA expression of NADPH P47-phox of the early-onset preeclampsia in the notch group were significantly higher than that of late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and the control group(P<0.01). The mRNA expression of p38MAPK-α of the early-onset preeclampsia in the notch group were significantly higher than that of late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and the control group (P<0.01). The mRNA expression of COX-2 of the early-onset preeclampsia in the notch group were significantly higher than that of late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and the control group (P<0.01).(4)The protein expression of LCHAD in the placenta of early-onset preeclampsia in the notch group, late-onset preeclampsia in the notch group and gestational hypertension in the notch group were significantly lower than that of the control group (P<0.01); and the protein expression of LCHAD in the placenta of early-onset preeclampsia in the notch group was significantly lower than that of late-onset preeclampsia in the non-notch group (P<0.01). The protein expression of NADPH P47-phox in the placenta of early-onset preeclampsia in the notch group was significantly higher than that of late-onset preeclampsia in the non-notch group and control group (P<0.05). The protein expression of p38MAPK-α in the placenta of early-onset preeclampsia in the notch group was significantly higher than that of late-onset preeclampsia in the notch group, late-onset preeclampsia in the non-notch group and control group (P<0.01). The protein expression of COX-2 in the placenta of early-onset preeclampsia in the notch group, late-onset preeclampsia in the notch group, gestational hypertension in the notch group, late-onset preeclampsia in the non-notch group, and gestational hypertension in the non-notch group, were significantly higher than that of control group (P<0.01).(5)The blood concentration of maternal FFA in the early-onset preeclampsia in the notch group was significantly negatively correlated with the mRNA and protein expression of placental LCHAD (r=-0.810,-0.932,P<0.01). There was no correlation between maternal TG level and the mRNA and protein expression of placental LCHAD in each group(P> 0.05).(6)The mRNA expression of placental LCHAD in the early-onset preeclampsia in the notch group was significantly negatively correlated with the mRNA expression of placental NADPH P47-phox and COX-2 (r=- 0.877,-0.762, P<0.05). The mRNA expression of placental LCHAD in the control group was significantly negatively correlated with the mRNA expression of placental COX-2 (r=- 0.565, P<0.01). The protein expression of placental LCHAD in the early-onset preeclampsia in the notch group was significantly negatively correlated with the protein expression of NADPH P47-phox (r=- 0.818, P<0.01). The protein expression of placental LCHAD in the control group was significantly negatively correlated with the protein expression of COX-2 (r=- 0.502,P<0.01). Conclusions The placental mRNA and protein expression of long-chain fatty acid oxidation enzymes were different in different clinical features of preeclampsia, which were reduced more obviously in the early-onset preeclampsia in the notch group than that of the late-onset preeclampsia in the notch group, and were negatively correlated with the elevated serum FFA level, significantly enhanced oxidative stress and inflammatory response, but with no correlation with serum TG level.

Objective To analyze the heterogeneous variation of serum free fatty acid (FFA) and lipids during early second trimester in women with or without uterine artery notch in pre-eclampsia (PE).Methods This is a prospective cohort study of 4 000 women with singleton pregnancies registered in early pregnancy and in whom regular check-ups were performed in Haidian Maternal & Child Health Hospital.Blood specimens were collected at gestational age 14-18 weeks at the same time of screening for Down's syndrome.One hundred and one cases with early diastolic notch of the uterine artery were included in the N+ group,and 172 cases without notch but at high risk of PE were included in the N-group at 22-24 weeks.In addition,205 women who were selected randomly at a ratio of 1 ∶ 5,without notch or PE high-risk factors,were also included in the N group.Both groups were subgrouped according to the outcomes of pregnancy complications:early-onset PE group EPE,late-onset PE (LPE),gestational hypertension (GH) group,gestational diabetes mellitus (GDM) group with normal blood pressure,and no complications (NC) group.The variation in FFA and other lipid metabolism indicators in the PE subgroups were compared and analyzed by two independent-sample t-test,one-factor analysis of variance,Chi-square test (or Fisher's exact) and Logistic regression.Results History of PE and pre-hypertension at first visit differed significantly between the N+ and N-groups [3.9％ (4/101) vs.0.8％ (3/377),x2=5.52,P＜0.05; pre-hypertension at first visit,42.2％ (43/101) vs.25.7％ (97/377),x2=10.91,P＜0.05].In the N+ group,23.8％ (n=24) of women had PE,of which 37.5％ (n=8) were early onset.In the N group,2.1％ (n=8) had PE,and all were late onset.The incidence of PE differed significantly between the N+ and N-groups (x2=59.72,P＜0.05).In the N+ group,FFA gradually decreased among the ePE,IPE,GH and NC groups [(0.68±0.27),(0.58±0.21),(0.57±0.21) and (0.49±0.19) mmol/L,F=2.78,P＜0.05]; Multivariate regression analysis showed that FFA (OR=135.68,95％CI:3.78-4 873.00) and PE history (OR=123.25,95％CI:9.27-i 638.00) were risk factors of ePE.Pre-hypertension at registration (OR=4.69,95％CI:2.08-10.58) and pre-pregnancy body mass index (BMI) 24-28 (OR=3.69,95％CI:1.26-10.83) were risk factors ofGH.FFA (OR=9.08,95％CI:2.49-33.01) and pre-pregnancy BMI ≥ 28 (OR=5.08,95％CI:2.16-11.92) were risk factors for GDM.Conclusions Serum FFA and TG levels in early second trimester are correlated with PE,especially the early-onset PE.The onset of PE is heterogeneous and affected by many factors,and occurs in patients with or without early diastolic notch of the uterine artery in the second trimester.Patients with notch are more likely to have early-onset PE,which is correlated with blood FFA and TG levels.

Advanced schistosomiasis is the most serious clinical type of schistosomiasis. Its diagnosis and treatment are relat?ed to many special departments,such as gastroenterology,general surgery,neurology,endocrinology,radiology,traditional Chinese medicine,blood purification,endoscopy,intervention,and ICU. It is necessary to apply a multidisciplinary treatment (MDT)mode. However,the mode has no universal standard and guide in practice. It is very important for the implementation of MDT mode of advanced schistosomiasis to form a treatment expert team,formulate the formal working procedures,and standard?ize the treatment schedules. The standardized implementation of MDT mode will be important to provide a more effective clinical decision on advanced schistosomiasis.

OBJECTIVETo explore the molecular basis for an individual with a rare weak D phenotype.

METHODSSerological methods were used to characterize the RhD blood group phenotype. The exons of RHD gene were amplified with PCR and sequenced. The presence of Rhesus box was tested by PCR to determine the homozygosity of RHD gene.

RESULTSThe RhD blood group of the proband was detected as weak D. The 10 exons of the RHD gene and Rhesus box could be amplified by PCR, and the genotype of RHD alleles was determined as RHD+/RHD-. The exons of the RHD gene were sequenced, and a 365C>T mutation in exon 3 was detected. Therefore, the RhD blood group of the proband was confirmed as weak D type 54 by both serological methods and DNA sequencing.

CONCLUSIONA weak D type 54 has been detected. A 365C>T mutation in RHD gene is responsible for the low expression of D antigen.

Alleles ; Base Sequence ; Exons ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phenotype ; Point Mutation ; Rh-Hr Blood-Group System ; genetics

OBJECTIVE: To study rare para-Bombay blood type Bm METHODS: ABO and H phenotype of the proband and her pedigree were determined with serological methods. The ABO genotype was analyzed by polymerase chain reaction-sequence specific primer(PCR-SSP). The full coding region of alpha-l,2 fucosyltransferase (FUT1) gene of the pedigree was analyzed by polymerase chain reaction and direct sequencing of the amplified fragments. The haplotype of the FUT1 gene were analyzed by cloning sequencing. RESULTS: The rare para-Bombay blood type Bm CONCLUSION Two new alleles of FUT1 gene (h
ABO Blood-Group System/genetics* ; China ; Female ; Fucosyltransferases/genetics* ; Genotype ; Humans ; Phenotype

OBJECTIVETo explore the molecular mechanism for a case with para-Bombay phenotype caused by α-1,2-fucosyltransferase (FUT1) gene mutations.

METHODSBlood phenotype of the propositus was determined by standard serological testing. Polymerase chain reaction-sequence specific primer (PCR-SSP) and direct sequencing of PCR product were used to analyze its ABO genotype. The PCR product of FUT1 gene was sequenced and analyzed.

RESULTSThe phenotype of the propositus was initially detected as para-Bombay A type. Direct sequencing of ABO gene showed that the genotype of the proband was A101/O01 (261G/del), which was consistent with the result of PCR-SSP. Two homo-mutations, 35C>T and 658C>T, were detected in the FUT1 gene by sequencing, and the genotype was determined as h(35T+658T)/h(35T+658T).

CONCLUSIONh(35T+658T)/h(35T+658T) is responsible for the para-Bombay phenotype of the propositus. The genotype is rare even in para-Bombay populations.

ABO Blood-Group System ; genetics ; Base Sequence ; DNA Mutational Analysis ; methods ; DNA Primers ; Fucosyltransferases ; genetics ; Genotype ; Homozygote ; Humans ; Male ; Phenotype ; Point Mutation ; Polymerase Chain Reaction

OBJECTIVETo identify and confirm a novel HLA allele in a Chinese Han individual.

METHODSHLA typing was performed by polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP) for HLA-A, -B and -DRB1 in a registered donor of China Marrow Donor Program(CMDP). Sequencing-based typing (SBT) was carried out to further confirm the novel allele of HLA-DRB1.

RESULTSThe SSOP result showed HLA-DRB1*09:03,15GEP, but an unusual pattern that could not be defined indicated potential presence of a novel allele. The SBT result showed the novel sequence has 1nt change from its closest allele DRB1*09:01:02 at nt306 where C to T(codon 73 GCC to GCT) resulting in no amino acid change. 73A was not changed.

CONCLUSIONA novel HLA allele, HLA-DRB1*09:01:07, has been identified and named officially by WHO Nomenclature Committee for Factors of the HLA System.

Base Sequence ; Blood Donors ; HLA-DRB1 Chains ; genetics ; Histocompatibility Testing ; methods ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA