Objective To study Ax subgroup’s molecular characteristics in Chinese Han population. Methods Eight samples suspected as Ax subgroup were analyzed and duplex PCR RFLP test was used to determine the primary ABO genotypes. These samples were then analyzed by another PCR RFLP test to identify whether there was an nt646 “T” to “A” mutation within the exon 7 of ABO gene, which was a known mutation related to most Ax phenotypes. Samples with discrepancy between serological and gene typing were chosen for further T A cloning and sequence analysis. Results Four out of all tested samples had the known nt646 “T” to “A” mutation. An A *weak01 allele including nt407 and nt467 “C” to “T” mis sense mutation was detected in this study. Moreover, a novel Ax allele with a new single nucleotide C to T mutation was detected at nt745. Another 2 unrelated samples were suspected as AxB through serological test, both of which contained higher quantities of anti A and showed strong agglutination with anti H. And their initial genotypes were BO, and sequence analysis clarified that both had normal O gene and novel nt640 “A” to “G” mutation in their B alleles. Conclusion The novel Ax alleles, one kind of novel B(A) allele and one A *weak01 allele in Chinese Han individuals,have been detected. B(A) phenotypes should have their molecular biology bases as well as other ABO subgroups.

Objective To investigate reliability and validity of the Chinese version of the schedule for assessment of insight (SAI).Methods The original version of SAI was translated into Chinese and then backtranslated into English to develop the Chinese version of SAI which reliability and validity were tested.220 subjects who met the diagnosis of Schizophrenia,schizophreniform disorder,schizoaffective disorder,delusional disorder or brief psychotic disorder were assessed.Results The Cronbach's α was 0.890 ; the test-retest reliability was 0.742 ; the inter-rater reliability was 0.986.Three main factors were obtained from the factor load matrix as awareness of illness,ability to relabel psychotic experience,compliance with treatment.The loading coefficient were 0.883-0.944.The correlation of the SAI total score and three subscales with insight of PANSS were-0.635,-0.305,-0.631 and-0.512 respectively (P ＜ 0.01).Conclusion The SAI has good reliability and validity.The scale is useful to measure insight in psychosis.

Objective To explore the effects of using proton pump inhibitors (PPIs) on the outcomes of hip fracture.Methods Searches were conducted through Medline,Embase,Cochrane Library and Chinese Biomedical Literature Database to identify the studies of the association between PPIs exposure and hip fracture.Quality of studies was assessed using the Newcastle-Ottawa Quality Assessment Scale.Pooled odds ratios (ORs) and 95％ confidence interval (CIs) were calculated for the risk of hip fracture associated with current exposure of PPIs.And several subgroups were analyzed by dosing duration,dose,osteoporosis and corticosteroid usage to explore potential study heterogeneities.All statistical analyses were performed with STATA software.Results Among 11 publications included for final analysis,there were a total of 1 107 577 subjects with an average age of over 60 years.A positive relationship existed between PPIs exposure and hip fracture with an OR of 1.46 (95％ CI:1.26-1.70,P =0.000) as compared with nonPPI-users,especially those on concurrent corticosteroid and PPIs.A significantly increased risk of hip fracture was found in the group of a short-term duration for under 1 year (OR =1.18,95％ CI:1.01-1.38,P =0.041),medium-term for 1-3 years (OR =1.23,95％ CI:1.01-1.49,P =0.038) and longer duration for over 6 years (OR =1.38,95％ CI:1.27-1.50,P =0.000).Furthermore,concurrent use of PPIs was not associated with an increased risk of hip fracture in a definite dose-response manner.As compared with non-PPI-users,no significantly increased risk of hip fracture was found in PPI-users with osteoporosis (P ＞ 0.05).Publication bias was not present.Conclusions Use of PPIs may be somewhat associated with an increased risk of hip fracture.Considering potential adverse effects,clinicians should prescribe cautiously PPIs for high-risk patients,especially elders.

Objective To investigate the spatiotemporal expression patterns of transforming growth factor-β(TGF-β) receptors type Ⅰ and type Ⅱ and their relationships with development of outflow tract(OFT) in mouse embryonic heart. Methods Serial sections of mouse embryos from embryonic day 9 (E9d) to embryonic day 14 (E14d) were stained using PAP immunohistochemical methods. Results Expressions of TGF-β receptors type Ⅰ (TGF-βRⅠ) and type II(TGF-βRⅡ) in the myocardial wall of OFT started at E10d, reached the reflection with splanchnic epithelium on the dorsal wall of the pericardial cavity at E11d. At E12d, expression intensity of TGF-βRⅠ and TGF-βRⅡ in myocardium increased to its highest level, and TGF-βRⅡ positive mesenchymal cells in OFT ridges could be detected. After E13d the staining intensity of TGF-βRⅠ and TGF-βRⅡ decreased rapidly,and at E14d,their expressions had fallen at the lowest.Conclusion The expressions of TGF-βRⅠ and TGF-βRⅡ in OFT are confined to the period of E10d to E14d, they may play important roles in regulating the myocardial cell proliferation, remodeling and septation of OFT, and promoting the differentiation from mesenchymal cells in the secondary heart field into smooth muscle cells in the distal end of OFT.

Objective In order to evaluate that whether Pluronic P85 coated poly(butylcyanoacrylate) nanoparti-cles was able to deliver antiepileptic drug phenytoin into the brain va bypassing mesial temporal lobe epilepsy (MTLE)-induced Pgp in a rat model of MTLE. Methods The rat model of MTLE, induced by li-pilocarpine, was divided in-to two groups (6 for nanoparticle drug group and 7 for PHT drug group). Immunohistochemistry assay was performed to detect Pgp expression at the hippocampus. Nanoparticles were prepared by interfacial polymerization method. Dialysate samples of brain were collected at 30, 60, 120, 180, 240 and 300 min after drug administration by microdialysis tech-nology. Samples were analyzed by high performance liquid chromatography (HPLC). Results The area under the curve (AUC) ratio of brain/plasma in Nanoparticle drug group was 0.370.10 which was significantly higher compared with 0.190.06 in conventional PHT drug group (P<0.05). The Pgp immunopositive area, as assessed by analysis of labeled surface area, was higher in the DG, CA3 and CA1 sector in the hippocampus of MTLE rats when compared to the normal rats. Conclusions Pluronic P85 coated PBCA nanoparticles can significantly deliver PHT into brain via bypassing MTLE-induced Pgp in a rat model of MTLE.

Objective To investigate the change of adiponectin (AD),tumor necrosis factor-alpha (TNF-α) and S100 levels in serum elderly patients with rheumatoid arthritis and cerebral infarct in order to evaluate the therapeutic effect of dioscornin.Methods One hundred patients with rheumatoid arthritis and cerebral infarct were selected as our subjects,who were hospitalized in the Department of Neurology of Recovery Changzhi Municipal People's Hospital and the Department of Internal Medicine of the Affiliated Hospital of Shaoxing Medical College,between 2006 September and 2010 September.All subjects (male 55,female 45,average age of 57 years old) were randomly divided into regular and dioscomin groups,50 for per group.Patients in regular group were treated with routine therapy and patients in dioscornin groups were treated with dioscornin 80mg,three daily plus regular treatment drug.Meanwhile 40 middle patients with single rheumatoid arthritis subjects were severed as controls.The changes of BMI,fasting plasma glucose,lipid factors,insulin sensitivity index (ISI),serum adiponectin,TNF-α and serum S100B were determined at treatment before and 6 months after treatments.Results The level of TNF-α,serum S100 in ERA patients were significantly higher andAdiponectin significantly lower than that of the control group,(TNF-α:[(89.0 ± 25.3) ng/L,(88.0 ± 24.2)ng/L vs(74.0 ±21.0) ng/L,F =3.292,P ＜0.05],[S100B:(0.102 ±0.051) μg/L,(0.101 ±0.045) μg/L vs(0.092 ± 0.031) μg/L,F =2.792,P ＜ 0.05],and AD and BMI were lower [AD:(7.2 ± 1.4) μg/L,(7.3 ±1.4) μg/L vs (18.1 ± 3.5) μg/L,F =17.057,P ＜ 0.01],[BMI:(18.9 ± 2.4) kg/m2,(19.0 ± 1.9) kg/m2 vs (21.8 ± 1.8) kg/m2,F =6.147,P ＜ 0.01].There was a negative correlation between adiponect and TNF-α,S100B (r =-0.46,-0.52,P ＜ 0.01) and positive correlation between adiponect and BMI (r =0.44,P ＜0.01).The adiponectin level was significantly increased in patients for six months after dioscornin treatment than that of control group.[AD:(12.2±2.9) μg/L,(7.8 ±1.8) μg/L vs (18.0 ±4.3) μg/L,F=6.480,P＜0.01].The level of TNF-α and S100B significantly decreased than that of the control group,TNF-α:[(72.0 ±21.0) ng/L,(82.0±23.0)ng/L vs (68.0 ±20.0) ng/L,F =3.065,P ＜0.05],[S100B:(0.092 ±0.021)μg/L,(0.099 ±0.031) μg/L vs (0.091 ±0.029) μg/L,F=3.030,P＜0.05].Conclusion Dioscornin could ameliorate the prognosis through decreasing the levels of TNF-α and S100B,and increasing adiponectin level in patients with rheumatoid arthritis and cerebral infarct.

C mutation. All 8 samples displayed the B(A) phenotype. Their real genotypes were B(A)/O. Conclusion Three B(A) alleles in the Chinese Han population were detected. Two alleles,B(A)700,B(A)640 were reported previously. One novel allele B(A)641, was first identified in this study.

OBJECTIVETo study the frequency of rare blood group Lu(a-b-) phenotype in a population from Shanghai region, and to explore the molecular basis of Lu(a-b-) by detecting the Lu and Lu relative mediator gene EKLF/KLF1.

METHODSDonors from Shanghai region were screened for Lutheran blood group by monoclonal anti-Lub using serological methods. Individuals with Lu(b-) were determined Lua, P1 and i antigens. Fifteen exons of the LU gene and 3 exons of the EKLF/KLF1 gene for the identified Lu(a-b-) samples were amplified and sequenced.

RESULTSTen Lu(a-b-) donors were obtained from 44 331 donors from Shanghai region. No homozygous or heterozygous mutations were found in the LU gene, whilst 7 mutations in EKLF/KLF1 gene were identified in the 10 samples.

CONCLUSIONThe frequency of rare Lu(a-b-) blood group in Shanghai was approximately 0.02%, and all the individuals had an In(Lu) phenotype. The molecular basis of such samples may be related to mutations in the EKLF/KLF1 gene.

China ; ethnology ; Humans ; Kruppel-Like Transcription Factors ; genetics ; Lutheran Blood-Group System ; genetics ; Mutation ; Phenotype

OBJECTIVETo investigate the difference of the transcripts between reticulocyte and non-reticulocyte cells in human blood.

METHODSGenomic DNA, reticulocyte RNA and total RNA of K-, K+ and Kell-null(K0) were extracted, then PCR, reverse transcription-PCR(RT-PCR) and nested PCR followed by sequencing or cloning-sequencing were used to analyze the KEL gene mRNA exons 1-19 and exons 2-8. Four kinds of monoclonal antibodies were labeled to detect the expression of Kell glycoprotein on red cells or leukocytes with flow cytometry.

RESULTSIn reticulocyte, only one normal KEL transcript faithful to the genomic structure was found in all tested samples except K0 which had 4 different transcripts. Sequence analysis of exons 2-8 of total RNA confirmed the alternative KEL transcripts existed in different samples, mostly caused by abnormal splicing, among them, skipping of exon 3 and a 16 bp insertion of intron 6 at the beginning of exon 7 were the most frequent. Although only one band was observed after amplifying the exons 1-19 from total RNA, the sequencing result showed it was a mixture of different sequences. There was strong expression of Kell glycoprotein on red cells except K0, but no or low expression on leucocytes by flow cytometry.

CONCLUSIONAlternative transcripts of KEL gene exist in different cells, which would be responsible for different Kell glycoprotein expression patterns on different cells. This study suggested that reticulocyte RNA was more suitable than total RNA for molecular study of KEL gene transcription.

Base Sequence ; Cloning, Molecular ; DNA ; genetics ; Exons ; genetics ; Genome, Human ; Genomics ; Humans ; Introns ; genetics ; Kell Blood-Group System ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; blood ; genetics ; Reticulocytes ; cytology ; metabolism ; Sequence Analysis, DNA

Objective:To explore the effects of early-life maternal deprivation on depressive-like behavior and neurogenesis in the granular layer of hippocampus in adolescent rats (6-7 weeks old).Methods:Neonatal rats were randomly divided into maternal deprivation group and control group, with 3 litters in each group.Rats in the maternal deprivation group were given maternal deprivation from 1 to 14 days after birth and rats in the control group were caged with the mother rats and raised normally.The body weight of rats at 5-6 weeks old was recorded and the increased body weight was calculated.When the rats were 6 weeks old, the sucrose preference test was carried out.Then the rats were killed and immunofluorescence histochemistry was applied to compare the expression of Ki67 and Nestin positive cells in the dentate gyrus of hippocampus.SPSS 22.0 software was used for statistical analysis.The data of the two groups were tested to conform to the normal distribution, and then t-test was carried out. Results:There was significant difference in body weight growth between the two groups at the age of 5-6 weeks.Compared with the control group, rats in the maternal deprivation group had lower body weight growth ((20.57±2.19) g, (30.57±1.25) g, t=3.96, P<0.01)) and lower sucrose preference rate((58.38±53.14)%, (73.88±3.67)%, t=3.21, P<0.01). The results of immunofluorescence showed that the number of Ki67 positive cells in the granular layer of hippocampus in the maternal deprivation group was less than that in the control group ((5.13±0.31), (7.60±0.38), t=5.09, P<0.01), and the number of Nestin immunofluorescence positive cells was more than that in the control group ((16.65±0.79), (7.64±0.70), t=8.51, P<0.01). The Nestin immunofluorescence positive cells in the maternal deprivation group had more protrusions and branches, and the morphology was similar to astrocytes, while the immunofluorescence positive cells in the control group had fewer protrusions, and the cell body was oval. Conclusions:Early-life maternal deprivation leads to depressive-like behavior in adolescent rats, which may be associated with the decrease of neurogenesis and activation of astrocytes in the dentate gyrus of the hippocampus.