Objective To study the association of peptidylarginine deiminase 4 (PADI4) (padi4_94 and padi4_104) genetic single nucleotide polymorphism (SNP) and rheumatoid arthritis (RA) susceptibility in Han population of Hebei province. Methods This hospital-based case-control study included 115 RA patients and 106 healthy controls. All the individuals were recruited from Han Hebei residents and were randomly selected. The genotype and allele frequencies of PADI4 gene polymorphisms (padi4_94 and padi4_104) were analyzed by PCR-DNA sequencing method. Results The distribution of padi4_94 and padi4_104 genotypes between the two groups was not significantly deviated from that expected by Hardy-Weinberg equilibrium (P>0.05). The combined effect of padi4_94 and padi4_104 SNPs was analyzed and three haplotypos (AA, AG and GG) were found but without GA haplotype. The genotype and allele distribution of padi4_94 and padi4_104 in the patients with RA was not significantly different from that in healthy controls,and the analysis of haplotypes revealed that no haplotypes were risk factors for RA. Conclusion In Han population of Hebei province, the SNPs of PADI4 (padi4_94 or/and padi4_104) is no associated with RA susceptibility. Therefore, it should not be regarded as a genetic risk factor for RA.

Objective In order to make a epileptic model in organotypic slice culture and to further investigate the epileptic pathology. Methods Immunohistochemistry, Fluoro-Jade B staining and BrdU labeling were carried out in the hippocampal slices with domoic acid treatment after 7 days of culture.Results Domoic acid could induce a series of pathological changes, such as, dispersion of granule cells(0.105±0.063) mm vs (0.057±0.012) mm, t = 4.8, P<0.01), neurogenesis in granular layer and cell loss of pyramidal cell and Mossy cell as well. Conclusions In hippocampal organotypic slices, domoic acid induced such pathological changes as human temporal lobe epilepsy and epileptic animal model. It is an ideal epileptic model for pathological, pharmacological and electronic physiological studies with great applicable prospect.

Objective To study the gene polymorphism at position padi4_94 in Chinese Han people of Hebei Province and the characters of distribution.Methods 106 healthy individuals and 115 rheumatoid arthritis patients in Hebei Province were randomly selected. Padi4_94 gene was screened by sequencing of the promotor. The correlations between the SNP site and anti-cyclic citrullinated peptide antibody (anti-CCP) was discussed.Results There were three genotypes: AA, AG and GG in Chinese Han people of Hebei Province. No significant differences of gene frequency and allele gene frequency were found between healthy individuals and RA patients, and no correlation between SNP and the production of anti-CCP.Conclusions Hebei Province The gene polymorphisms at position padi4_94 existed in Chinese Han people of Hebei Province,but possibly would not affect RA susceptibility and the production of anti-CCP.

Objective To investigate the growth and expression of PDGF of VSMCs in response to stimulation of autologous serum and mechanical injury. Methods An vitro model simulating the condition as possible as that after PTA.PDGF of every medium sample from every group was detected by ELISA,and the values of MTT of every cellular sample was measured by MTT to show the growth and proliferation of every group. Results After stimulation by autologous serum and mechanical injury,SMCs of the experimental group showed the value of MTT increasing,but SMCs in control group reached on 3rd day.At the same time,the expression of PDGF also increased gradually,obtaining peak gradually up to peak on day 4/5 nearly 2.0-fold as much as that of SMCs in the control group. Conclusions After on the 5th day,stimulation with autologous serum and mechanical injury,VSMCs of rabbit showed the stronger ability of growth/proliferation,and autocrine of PDGF also increased gradually,reaching peak on 4-5 d,probobly simulating to those in vivo.

ObjectiveIn order to compare the alteration of reelin-immunoreactive Cajal-Retzius cells (CR cells) in molecular layer of dentate gyrus of APPswe transgenic mice with wild type, the histochemical and developmental characteristics of CR cells were studied, therefore, the roles of CR cells in Alzheimer's disease would be revealed further.Methods The Thioflavine S staining, reelin immunofluorescence with or without reelin/glutamate and reelin/GABA immuno-double staining were carried out in the study. In the meantime, Western blotting was used to study the expression of reelin in hippocampi of the both wild type and transgenic mice. Results Reelin positive CR cells could be double-labeled with either glutamate or GABA immunostaining. Caspase-3 immunofluorescence demonstrated that some CR cells went through apoptosis during their development. Compared with wild type, CR cells in APPswe transgenic mice had significantly decreased in the molecular layer of the dentate gyrus. The result was supported with Western blotting analysis of reelin expression in hippocampus. Conclusion Reelin could be co-expressed with either glutamate or GABA, suggesting CR cells would be glutamatergic exciting neurons and GABAergic interneurons. The loss of CR cells during development probably was caused by the neuroapoptosis. Significant decrease of CR cells in hippocampus of APPswe transgenic mice indicated reelin may play an important role in AD pathological alterations.

BACKGROUND:Tumor stem cels are the root of cancer recurrence and metastasis, so clinical researches should focus on the effects of different treatments on tumor stem cels.
OBJECTIVE:To explore the effects of endocrine therapy and chemotherapy on stem cels in patients with breast cancer.
METHODS:After recovery and cultivation of estrogen receptor-positive human breast cancer cel lines MCF-7, passage 3 cels in logarithmic phase were selected and divided into three groups containing control, estradiol and estradiol with tamoxifen groups. The estradiol group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol was added into the medium, respectively; the estradiol with tamoxifen group was divided into three subgroups: 10-7, 10-8 and 10-9 mol/L estradiol with 10-6 mol/L tamoxifen were added into the medium, respectively. The same amount of absolute ethyl ethanol was added into the medium of control group. Fifteen female patients with late recurrence and metastasis of breast cancer received chemotherapy as recurrence and metastasis group. Another 15 healthy volunteers were selected as healthy control group.
RESULTS AND CONCLUSION:The proportion of CD44+CD24-/lowcel subsets in the estradiol and estradiol with tamoxifen groups was significantly higher than that of the control group (P < 0.05), and the proportion of CD44+CD24-/low cel subsets in the estradiol group was significantly higher than that of the estradiol with tamoxifen group at the same concentration (P< 0.05). The proportion of CD44+CD24-/lowcel subsets had no significant differences among groups at 10 and 20 days of culture (P < 0.05). The proportion of CD44+CD24-/low cel subsets significantly increased in MCF-7 cels after 24-hour intervention with different chemotherapy drugs. But only the proportion of CD44+CD24-/low cel subsets in the paclitaxel and doxorubicin groups was significantly higher than that of the control group after 20-day intervention (P < 0.05). Besides, the proportion of CD44+CD24-/low cel subsets in the peripheral blood of healthy volunteers was significantly lower than that of the recurrence and metastasis group (P < 0.05). Among 15 patients with late recurrence and metastatic of breast cancer, 9 had stable disease, 5 had partial remission, 1 had failed chemotherapy and cancer progression. Moreover, the proportion of CD45-CD44+CD24-/low cel subsets in the peripheral blood of patients sensitive for chemotherapy was significantly lower than that before treatment (P < 0.05). In conclusion, both endocrine therapy and chemotherapy exert a certain effect on the CD44+CD24-/low cel subsets of breast cancer positive for estrogen receptor. Given that CD44+CD24-/low cel subsets in MCF-7 cels resist chemotherapy drugs, the proportion of CD45-CD44+CD24-/low cels in the peripheral blood of patients sensitive for chemotherapy is decreased.

Objective:To investigate the improved superantigen activity of SEC2(T20L/G22E) compared with recombinant staphylococcal enterotoxins C 2 ( rSEC2 ).Methods: The proliferation of spleen lymphocytes and T-cell subpopulations induced by rSEC2 and SEC2(T20L/G22E) were examined by WST-1 and flow cytometry separately,and the gene expression of cytokines and Vβspecificities were quantified by real-time PCR.Results: WST-1 and Flow cytometry assays showed that the superantigen activity of SEC2(T20L/G22E) was improved due to enhanced T-cell stimulating potency,resulting in massive activation of T-cells,particularly CD4+and CD8+T-cells.Quantitative real-time PCR assay showed that despite similar Vβspecificities induced by rSEC 2 and SEC2 (T20L/G22E),the quantities of activated T-cells bearing specific Vβwere different,and SEC2(T20L/G22E) could stimulate more gene expression of associated cytokines simultaneously.Conclusion: The results strongly suggested that the increased SEC 2 ( T20L/G22 E)-TCR-binding affinity contributed to more T-cells activation and cytokine release ,which elicit powerful immune activition.

Objective To investigate the role of Zn 2+and Zn2+-coordinating glutamic acid residues ( 71Glu and 80Glu ) in the superantigenic activities of staphylococcal enterotoxin C2 ( SEC2 ) .Methods Over-lap PCR was used to amply genes encoding recombinant mutant proteins rSEC 2 ( E71A), rSEC2 (E80A) and rSEC2 (E71A/E80A).The mutant proteins were expressed in E.coli BL21 (DE3) and puri-fied by affinity chromatography .The differences of biological activities between rSEC 2 and its mutants were compared in vitro.The effects of Zn 2+on the superantigenic activities of rSEC 2 were evaluated by analyzing the proliferation of splenic lymphocytes in the presence or absence of ethylenediaminetetraacetic acid ( EDTA) .Results The substitution of glutamic acid residue at position 71 and 80 by alanine residue had no significant effects on the superantigenic activities and the conformational stability of rSEC 2 mutants.How-ever, traces of Zn2+(10μmol/L) could significantly enhance rSEC2-induced proliferation of splenic lympho-cytes, and only certain amount of Zn 2+could completely restore rSEC2-induced proliferation in the presence of EDTA.Conclusion This study indicated that mutations at Zn 2+-coordinating glutamic acid residues had no significant effects on the conformational stability and the superantigenic activities of SEC 2.Zn2+might play a role in regulating the superantigenic activities of SEC 2 through some indirect ways .

Objective To explore and compare the clinical effects of pregnant women with liver cirrhosis after hepatitis in cesarean section between general anesthesia and single spinal anesthesia.Methods Clinical data of pregnant women of 35,including compensated cirrhosis 26 cases,decompensated cirrhosis 9 cases,aged 24-45 years,weighing 55-98 kg,falling into ASA Ⅱ or Ⅲ,were divided into group A (general anesthesia group,n=15) and B (single spinal group,n=20).In patients of group A,propofol were infused at 1.5 mg/kg till the time for fetal delivery,after intubation,followed by infusion of remifentanil at a rate of 0.2 μg·kg-1·min-1 TCL combined with propofol until the end of operation.In group B,the puncture was performed at L3-4 interspace,following by intrathecal injection of ropivacaine 10~15 mg and within 10 seconds.With liver cirrhosis after hepatitis were collected and analyzed,retrospectively.Preoperative general status,blood loss and time of operation,Apgar score of neonates compared and analyzed in groups A and B.And the changes of hepatic function and coagulation function of the body under different anesthesia methods were also studied.The influential factors of postoperative elevation of hepatic function were analyzed by non-conditional logistic regression models.Results Blood loss and time of operation,Apgar score of neonates and postoperative hospital stay had no significant differences between the two groups.Postoperative albumin of group A was higher than that of preoperative,with a statistical significance (P<0.05).Alanine aminotransferase,aspartate aminotransferase of postoperative of group B were increased,with a statistical significance (P<0.01).Logistic analysis showed that the type of anesthesia was the possible influencing factor of postoperative elevation of total bilirubin (OR=12.04,95%CI 1.14-127.45).Conclusion The negative effect of single spinal anesthesia on hepatic function more than that of general anesthesia in pregnant women with liver cirrhosis in cesarean section.