Objective In order to study the role of APP SW mutation in the aetiology of Alzheimer′s disease(AD), the phenotypes of the PDAP SW transgenic mice were investigated. Methods Body weight, coat color and reproducibility of transgenic mice were observed;The pathological changes in the brain of the transgenic mice were examined using immunohistochemistry; Behavioral changes of transgenic mice were examined by Y-maze. Result At 3 month′s age, the mean body weight of the transgenic mice was (31.0?3.7) g, that of non-transgenic was (34.0?2.9) g;while the mean body weight of F1 transgenic mice was (32.0?3.3) g, of negative mice was (31.0?4.2) g. There were no significant difference between these two groups. There were three transgenic mice with abnormal coat and one male was infertile. The transgenic mice had amyloid deposit in their brains. In the Y-maze test, transgenic mice showed an increased number of arm entered (53?7 versus 37?4) and an impaired spontaneous alternation. The frequency of alternation was 48.2% in the transgenic mice and 76.4% in the non-transgenic mice,respectively.Conclusion The PDAP SW transgenic mice had typical pathological and behavioral changes similar to that of AD and could be used as an animal model for studying AD.

Preimplantation Genetic Diagnosis(PGD) is a pre-gestational diagnostic technique used to identify genetic defects in embryos created through in vitro fertilization prior to transfer into the uterus.It is currently the only option available to avoid the dilemma of a pregnancy termination due to unfavorable prenatal diagnosis of high risk genetic diseases,however,the limitation of this technique could result in uncertain results.To avoid ethics disputes,patients must undergo a medical informed consent process to fully understand the risks prior to undergoing PGD.This process is often hampered by the patients' restricted recognition ability.Here we discuss possible strategies to help patients completely comprehend the critical issues relating to PGD,including comprehensiveness of PGD related information,and all possible benefits and risks of currently available operational and detection techniques.Based on this, It is necessary to sign a detailed medical informed consent according to different inherited disease.

Objective To study the security in HBV carried infertility patients during the in-vitro-fertilization procedure. Methods Serologic testing of HBV infection (HBsAg, HBsAb, HBeAg, anti-HBe, anti-HBc-IgG) of blood, follicle fluid/sperm and fertilization culture medium/post-washing sperm in female/male carried patients were detected by ELISA on the day of oocytes collection. Results In 18 female patients who were seropositive for HBsAg, HBeAg and anti-HBc-lgG, the same antigen and antibody could be detected in all 18 folli-cle-fluid and fertilization culture medium(100%, 100%). In 131 female patients who were sernpesitive for HBsAg, anti-HBe, anti-HBc-IgG, the same antigen and antibody could be detected in 84 follicle-fluid(64. 1%), and 8 fertilization culture medium(6. 1%). If the patients'follicle-fluld was negative for all the markers, their fertilization culture medium remained negative. In 23 male patients who were sero-positive for HBsAg , HBeAg,and anti-HBc-lgG, the same antigen and antibody could be detected in only 6 sperm (26. 1%), all the postwashing sperm were exhibited negative. In 121 nude patients who were seropositive for HBsAg, anti-HBe, anti-HBc-lgG, the same antigen and antibody could be detected in only 7 sperm (5. 8%), and all the post-washing sperm were negative. Conclusion In IVF-ET procedures, the risk of HBV transmission by follicle-fluid and fertilization culture medium in those female patients who showed serepesitive for HBsAg , HBeAg, and anti-HBc-lgG can not be decreased. The risk in those female patients who showed seropositive for HBsAg, anti-HBe, anti-HBe-lgG and male patients in IVF-ET procedures can be decreased.

Objective To explore the association between rs3179060C/A polymorphism in the ex-on 1 of TNF-a gene and hyperandrogenism and polycystic ovary syndrome (PCOS). Methods One hundred thirty PCOS women and one hundred seventy five normal women as controls were enrolled in this study. The genotypes were screened by polymerase chain reaction-On/off switch and the product was isolated by e-lectrophoresis on a 2. 5% agarose gel containing ethidium bromide and visualized using an ultraviolet transil-luminator. The relationship of TNF-a alleles to serum testosterone level was analyzed in PCOS patients. Results Three genotypes were identified, corresponding to CC, CA, AA, and two alleles were screened, corresponding to C and A. The frequencies of the CC, CA, AA genotypes were 58. 4% ,23.1% ,18.5% vs. 72. 0% , 17.7% , 10. 3% in PCOS group and control group, showing statistically significant difference between two groups( P < 0.05 ). The allelic frequency was 70.0% for the C allele and 30.0% for the A in P-COS group, and 80. 9% for the C allele and 19. 1% for the A in control group, respectively, showing statistically significant between two groups ( P <0.05). The relationship was not observed between rs3179060C/ A polymorphism and serum testosterone level in PCOS patients in Han Chinese racial origin ( P >0.05). Conclusions The TNF-a gene rs3179060C/A polymorphism may be a risk factor for the pathogenesis of P-COS in Chinese women, but it was not effect on hyperandrogenism in PCOS women.

Objective To derive hematopoietic stem cells with functional properties of hematopoietic reconstitution from murine embryonic stem (ES) cells. Methods ES-D3 cells by formation of the day-4 embryoid bodies (4dEBs) were induced into hematopoietic stem cells by co-culture with murine bone marrow endothelial cell-conditional medium (mBMEC-CM) and the fetal liver stromal cell-conditional medium (FLSC-CM). This experiment was designed to 4 groups (mBMEC-CM + FLSC-CM group, mBMEC-CM group, FLSC-CM group, and the control group). Results The total cell numbers, CD34 + cell numbers, and colony numbers formed in the mBMEC-CM + FLSC-CM group were the highest among the 4 groups. The cells in the mBMEC-CM + FLSC-CM group resumed the hematopoietic system of the mice after being transplanted with the inducing cells. Conclusion The culture condition combing mBMEC-CM with FLSC-CM can promote murine ES cells differentiating into hematopoietic stem cells with functional properties of hematopoietic reconstitution.

Objective　To establish the method of 2-dimensional electrophoresis(2-DE) for proteins of human spermatozoa and to construct a protein map of human spermatozoa. Methods　The sperm pellet was prepared with simple Percoll layer protocol. We studied the effects of various sample preparation methods, loading quantities and isoelectric-focusing protocols on the quality of silver-stained 2-DE map, and constructed a primary protein map of human spermatozoa. Result　Up to 703 protein spots were acquired with sample preparation Method Ⅰwhile only 194～210 spots with Method Ⅱ.With immobilized pH gradients and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(IPG-DALT) we could acquire over 700 spots while only 280～300 with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(ISO-DALT). Conclusion　It is satisfactory to lyse sperm with sample preparation Method Ⅰ and to separate sperm proteins by IPG-DALT for establishing 2-D map of human sperm.

Objective To analyze the chromosome abnormalities in the patients with primary and secondary amenorrhea in the infertility clinic, and provide the guidance for their reproduction.Methods Peripheral blood lyphocytes chromoseme samples were prepared and the karyotype was analyzed according to routine method.Results 24 abnormal chromosome karyotypes were detected in 108 cases, the abnormality detection rate was 22.2%, and it was mainly involved in the numerical or structural abnormality of chromosome X. Conclusions Chromosome abnormality is an important cause for primary and secondary amenorrhea, and the X chromosome abnormality is involed mainly.For those patients,chromosome examination should routinely be taken before assistant reproductive technique.

Objective To establish a system in which mouse embryonic stem cells (mESCs) differentiate into insulin-secreting cells in vitro. Methods mESCs were cultured to form embryoid bodies(EBs). EBs were cultured in serum-free medium to obtain nestin-positive cells. The selected nestin-positive cells were expanded,then added nicotinamide into the medium to induce the differentiation of nestin-positive cells. Immunochemistry staining and flow cytometry analysis were made on 15th day after induction.Results Flow cytometry analysis revealed that the percentage of nestin-positive cells were different from EBs. The percentage of nestin-positive cells from EBs were higher than those of other diameters. Nestin-positive cells induced by nicotinamide formed islet-like cell clusters. Percentage of insulin-positive cell induced by 10 mmol/L nicotinamide was higher than those induced by 0 mmol/L or 5 mmol/L nicotinamide(P

Objective:To explore the influence factors on amplification of single cell duplex-nested PCR.Methods:The mutational loci region CD41-42 and IVS-Ⅱ654 of ?-globin gene were amplified by duplex-nested PCR with different combination of primers concentration, different Taq DNA polymerases, different neutralization buffers and with or without predenaturation at 98 ℃ before the PCR amplification in single lymphocyte or single blastomere, thus, to investigate the influence of these factors on the amplification efficiency of PCR.Results:TaKaRa EX Taq was the most efficient Taq DNA ploymerase among different Taq DNA ploymerases; primer pair R1+F1 at final concertration of 0.25 ?mol/L and R2+F2 at 0.3 ?mol/L were the most efficient ones in amplification among different combinations of primers concentrations; the amplification efficiency in neutralization buffer-1 (200 mmol/L Tricine) was obviously higher than that of neutralization buffer-2 (900 mmol/L Tris-HCl, pH 8.3/300 mmol/L KCl/200 mmol/L HCl)(P0.05). Conclusion:There were remarkable differences of the amplification efficiency of single cell duplex-nested PCR while using different combination of primers concentrations, different Taq DNA polymerases, different neutralization buffers. However, predenaturation at 98 ℃ before the single cell PCR amplification could not improve the PCR amplification efficiency

Objective To compare the therapeutic effect of two Methodsof in vitro oocyte maturation (IVM) technology with polycystic ovary syndrome (PCOS) patients in infertility treatment.MethodsRetrospective analysis was performed on patients with PCOS infertility who were stimulated with two different kinds of stimulating method of the IVM cycle in our center during April 2010 and August 2010.There were 20 patients in the temporary FSH stimulating group (group A) and 31 patients in the HCG stimulating group (group B).The number of oocytes maturation rate,fertilization rate,cleavage rate,excellent embryo rate,implantation rate,pregnancy rate per oocyte recycle,pregnancy rate per transfer and abortion rate of transfer were compared.ResultsThere was no significant difference between the two groups of age,duration of infertility,body mass index,based sex hormone levels by t test(P＞0.05).There was no significant difference between the two groups of the number of oocytes retrieved,oocyte maturation rate,cleavage rate,and abortion rate of transfer(P＞0.05).In group B,the fertilization rate was 80.3%,good-quality-embryo rate was 49.8%,implantation rate was 31.5%,pregnancy rate per oocyte recycle was 54.8%,which was significantly higher than in group A (73.9%,35.4%,12.5% and 25.0%,respectively).ConclusionsIf IVM was used to treat PCOS patients with infertility,HCG stimulating will be better than FSH stimulating,and it could result in a higher clinical pregnancy rate.