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Biomedical subjects

Y Ji

Publications and source records attributed to Y Ji.

At least 109 records · Page 6Linked to original sources

Expressed copies of the MN7 (D15F37) gene family map close to the common deletion breakpoints in the Prader-Willi/Angelman syndromes.

Approximately 70% of patients with Prader-Willi syndrome or Angelman syndrome have a similar sized de novo deletion of 3-4 Mb in the proximal region of 15q. The distal breakpoints appear to cluster between the P gene (OCA2) and D15S24, whereas two deletion breakpoint clusters have been identified on the proximal side (one centromeric to D15S541 and one between D15S541 and D15S9). Based on the identification of a gene family in 15q11-->q13 (MN7, D15F37), we have previously proposed that the presence of multiple copies of this sequence may be related to the instability of this region. Using fluorescence in situ hybridization and YAC mapping, we have found that at least one D15F37 locus is centromeric to D15S9 and at least two are between OCA2 and D15S24. As determined by cDNA cloning and sequence analysis, each of the individual loci is expressed. The close proximity of the D15F37 loci and the deletion breakpoints suggests that the common deletions arise by unequal crossover events at or near these loci.

Adult↗

Co-expression of transforming growth factor beta and interferon tau during peri-implantation period in the ewe.

The transforming growth factor beta (TGFbeta) family is known to control cell migration, growth, differentiation, function and regulation of extracellular matrix, all of which are required for the process of implantation. Expression of TGFbeta by the conceptus and endometrium was studied during the period of implantation in the ewe. A total of thirty-four ewes were hysterectomized on day 12, 14, 16, 18 or 20 of pregnancy (day 0 = day of estrus). Conceptus (200 mg wet weight) and endometrial (300 mg wet weight) tissues were cultured in vitro in 7 and 10 ml Eagle's minimal essential medium, respectively. The culture media were subjected to a bioassay to determine concentrations of TGFbeta. Conceptus culture media (CCM) were also analyzed for contents of ovine interferon-tau (oIFNr), low molecular weight acidic protein, produced by the trophectoderm between days 8 and 21 of pregnancy. Whole uteri including conceptus(es) and conceptuses (day 16) only were fixed and subjected to immunohistochemical and in situ hybridization studies. Levels of oIFNr produced by conceptuses were the highest on day 16 at 4.4 microg/ml. Concentrations of TGFbeta in day 12, 14, 16, 18 and 20 CCM were 38+/-19, 102+/-56, 862+/-152, 728+/-191 and 336+/-106 pg/ml, respectively, and approximately 90% of TGFbeta activity in CCM was due to TGFbeta1 whereas less than 10% was due to TGFbeta3 based on neutralization with TGFbeta subtype-specific antibodies. Immunohistochemical studies revealed that day 16 conceptuses displayed major staining for TGFbeta1, no beta2 staining and minor staining for beta3. In situ hybridization studies also revealed that day 16 trophectoderm possessed most TGFbeta1 mRNA while day 14 trophectoderm and day 20 chorion/amnion displayed weaker staining for TGFbeta1 mRNA. TGFbeta in day 12, 14, 16, 18 and day 20 endometrial culture media was 156+/-37, 129+/-33, 49+/-22, 62+/-23 and 179+/-40 pg/ml, respectively, and approximately 65% and 35% of the activities were due to TGFbeta1 and beta2, respectively. These results indicate that TGFbeta production by the conceptus coincides with the time when oIFNtau production starts to decline. These observations support the postulate that TGFbeta may play an important role in implantation in the ovine species.

Animals↗

Long-term efficacy of recombinant interferon alpha 2a in the treatment of chronic hepatitis C: a randomized prospective study comparing two dose schedules in Chinese patients.

OBJECTIVE: To compare the long-term efficacy of a dose of 3 million units (MU) of r-IFN alpha 2a (IFN-alpha 2a) three times a week (t.i.w.) for 6 months with a starting dose 6 MU for 3 months and subsequent reduction to 3 MU t.i.w for further 3 months. METHODS: Sixty-eight serological and histological chronic hepatitis C patients with elevated serum alanine aminotransferase (ALT) were enrolled and randomized into two groups. Sixty-three patients were completed with full course of treatment. Five patients were withdrawn from trial (2 due to personal reasons and 3 due to adverse drug reactions during treatment). Thirty patients received 6 MU IFN-alpha 2a t.i.w., 3 months followed by 3 MU t.i.w. for another 3 months (Group A). Thirty-three patients received 3 MU IFN-alpha 2a t.i.w. for 6 months (Group B). RESULTS: The sex, age, baseline serum bilirubin, ALT and aspartate aminotransferase (AST) levels were matched in both groups. At the end of the 6th month, the complete and partial response rates in Group A were 60.0% and 16.7% respectively, and the clearance of serum HCV-RNA was 53.3%. In Group B, the complete and partial response rates were 72.7% and 6.1% respectively, and the clearance of HCV-RNA was 61.3%. The patients were followed up for 6, 12, and 18 months after stopping treatment. In Group A, the rates of complete normalization of ALT and clearance of serum HCV-RNA at 24 months were 50.0% and 60.0% respectively. In Group B, the rates of normalization of ALT and clearance of HCV-RNA at 24 months were 54.4% and 41.9% respectively. The efficacy between the two groups showed no statistically significant difference. The response rates of treatment were similar to those in the patients with HCV genotype 1b and 2a. Six patients (10.8% of the study population) developed neutralization antibodies to IFN-alpha 2a during treatment, and four of them were responded to the treatment. Adverse drug reactions (ADR), were common, but most of them were tolerable, and the incidence of ADR was in both groups, but the severity was higher in Group A. CONCLUSIONS: IFN-alpha 2a is effective in the treatment of Chinese patients with chronic hepatitis C. The sustained response rates and adverse drug reactions among two dose schedule groups are similar.

Adolescent↗

[Application of VNTR D17S30 locus polymorphism in the paternity test].

A sensitive and rapid PCR-based technique was adopted to genotype the VNTR D17S30 locus. It was confirmed through the genetic analysis of 20 normal families that the inheritance of D17S30 locus coincides with Mendelian law as simple co-dominant. Retrospective analysis of 100 paternity cases demonstrated that D17S30 locus could be used in forensic paternity test in our country. The exclusion probability estimated from allele frequencies of D17S30 locus (74.04%) does not differ significantly from the observed rate of exclusion (80.00%) in these cases. In all excluded paternity cases there are two in which the exclusion evidence is solely provided by the D17S30 locus.

Chromosomes, Human, Pair 17↗

[Primary studies on the establishment of cytomegalovirus murine model].

Intraperitoneal inoculation of Murine cytomegalovirus(MCMV) into Balb/C mice at age of 4 weeks resulted in acute MCMV infection of the mice. The infected mice had body weight reduction, growth retardation, salivary gland swelling and death. A lethal dose was 5 x 10(6) PFU/ml of MCMV. Virus could be recovered from salivary gland and the infectivity titer of MCMV reached to a high titer of 2 x 10(5) PFU/ml. Plaque formation of MCMV in 3T3/Swiss albino cells was identified. Histopathological examination showed the appearance of MCMV-specific intracytoplasmic inclusion bodies in neuron cells of the mouse brain.

3T3 Cells↗

[Purification and properties of inulinases from Aspergillus niger M89].

Four inulinase components were purified from Aspergillus niger M89 by (NH4)2SO4 fractionation, Sephadex G-200 gel filtration, DEAE-cellulose chromatography and polyacrylamide gel electrophoresis (PAGE). The molecular weight of EI EII EIII and EIV were indicated to be 102.6, 97.9, 62.5 and 36.5 kD respectively by SDS-PAGE. Measured by isoeleetric focusing, their isoelectric points were 4.15, 4.24, 4.48 and 4.15 respectively. Four forms of inulinase exibited maximal inulinase activity between 55-60 degrees C with optimal pH 4.0-5.0. There were some difference in their thermal stability. The little the molecule of the component was, the stronger its thermal stability was. All of four components were exo-acting inulinases.

Aspergillus niger↗

High affinity dimerization by Ski involves parallel pairing of a novel bipartite alpha-helical domain.

c-Ski protein possesses a C-terminal dimerization domain that was deleted during the generation of v-ski, and has been implicated in the increased potency of c-ski in cellular transformation compared with the viral gene. The domain is predicted to consist of an extended alpha-helical segment made up of two motifs: a tandem repeat (TR) consisting of five imperfect repeats of 25 residues each and a leucine zipper (LZ) consisting of six heptad repeats. We have examined the structure and dimerization of TR or LZ individually or the entire TR-LZ domain. Using a quenched chemical cross-linking method, we show that the TR dimerizes with moderate efficiency (Kd = 4 x 10(-6) M), whereas LZ dimerizes poorly (Kd > 2 x 10(-5) M). However, the entire TR-LZ domain dimerizes efficiently (Kd = 2 x 10(-8) M), showing a cooperative effect of the two motifs. CD analyses indicate that all three proteins contain predominantly alpha-helices. Limited proteolysis of the TR-LZ dimer indicates that the two helical motifs are linked by a small loop. Interchain disulfide bond formation indicates that both the LZ and TR helices are oriented in parallel. We propose a model for the dimer interface in the TR region consisting of discontinuous clusters of hydrophobic residues forming "leucine buttons."

Amino Acid Sequence↗

Scavenger receptor BI promotes high density lipoprotein-mediated cellular cholesterol efflux.

Scavenger receptor BI (SR-BI) binds high density lipoproteins (HDL) with high affinity and mediates the selective uptake of HDL cholesteryl ester. We examined the potential role of SR-BI in mediating cellular cholesterol efflux. In Chinese hamster ovary cells stably transfected with murine SR-BI, overexpression of SR-BI resulted in a 3-4-fold stimulation of initial cholesterol efflux rates. Efflux rates correlated with SR-BI expression in cells and HDL concentration in the medium. When incubated with synthetic cholesterol-free HDL, SR-BI-transfected cells showed approximately 3-fold increases in initial rates of efflux compared with control cells, indicating that SR-BI expression enhances net cholesterol efflux mediated by discoidal HDL. In six different cell types, including cultured macrophages, the rate of efflux of cholesterol mediated by HDL or serum was well correlated with cellular SR-BI expression level. In addition, in situ hybridization experiments revealed that SR-BI mRNA was expressed in the thickened intima of atheromatous aorta of apolipoprotein E knockout mice. Thus, SR-BI is an authentic HDL receptor mediating cellular cholesterol efflux. SR-BI may facilitate the initial steps of HDL-mediated cholesterol efflux in the arterial wall as well as later steps of reverse cholesterol transport involving uptake of HDL cholesterol in the liver.

Animals↗

Subunit specificity and organ distribution of glutathione transferase-catalysed S-nitrosoglutathione formation from alkyl nitrites in the rat.

Glutathione transferase (GST)-catalysed S-nitrosoglutathione (GSNO) formation from alkyl nitrites was determined with the homodimers 1-1, 2-2, 3-3, and 4-4 isolated from rat liver. The 4-4 isoform showed a high specificity for the alkyl nitrites. Total GST activities were studied in homogenates from different organs. The liver showed highest GST activity both with amyl nitrite and with 1-chloro-2,4-dinitrobenzene (CDNB) as substrate, the activity ratio of amyl nitrite over CDNB being 3.8. In lung and heart, these ratios were 6.2 and 5.7, respectively, indicating a selectivity of these organs for alkyl nitrite metabolism and GSNO formation.

Amyl Nitrite↗

Intranasal immunization with C5a peptidase prevents nasopharyngeal colonization of mice by the group A Streptococcus.

Early inflammatory events are initiated by phased production of C5a and interleukin-8 in tissue. Most serotypes of group A streptococci express a surface-bound peptidase (SCPA) which specifically cleaves mouse and human C5a chemotaxins. This study investigates the impact of SCPA on colonization of the nasopharyngeal mucosa of mice and evaluates its potential to induce protective immunity. Two strains, serotypes M6 and M49, which contain insertion and deletion mutations in the SCPA gene (scpA) and represent the two major subdivisions of group A streptococci, were characterized and compared in a mouse intranasal infection model. In this model, SCPA mutants were more rapidly cleared from the nasopharynges of inoculated mice compared with wild-type strains. A 2,908-bp fragment of scpA49 gene, obtained by PCR, was ligated to the expression vector pGEX-4T-1 and expressed in Escherichia coli. The affinity-purified deltaSCPA49 protein proved to be highly immunogenic in mice and rabbits. Although the purified deltaSCPA49 immunogen lacked enzymatic activity, it induced high titers of rabbit antibodies which were able to neutralize peptidase activity associated with M1, M6, M12, and M49 streptococci in vitro. This result confirmed that antipeptidase antibodies lack serotype specificity. Intranasal immunization of mice with the deleted form of the SCPA49 protein stimulated significant levels of specific salivary secretory immunoglobulin A (IgA) and serum IgG antibodies and reduced the potential of wild-type M1, M2, M6, M11, and M49 streptococci to colonize. These experiments suggest a new approach to vaccine development for prevention of streptococcal pharyngitis.

Adhesins, Bacterial↗

Effect of optic disc size or age on evaluation of optic disc variables.

AIMS/BACKGROUND: It has been reported that the number of optic nerve fibres decrease with age, and the cup/disc (C/D) ratio increases as the optic disc size increases. Consequently, the normal value of the optic disc variables measured by an optic disc analyser may change according to the optic disc size or age. The effect of individual variations in optic disc size or age on interpretation of optic disc variables was investigated. METHODS: Topographic optic disc variables of 104 normal Asian adults of both sexes aged 40 to 68 were measured using a confocal scanning laser ophthalmoscope (TopSS, Laser Diagnostic Technologies, Inc). Fourteen variables were evaluated according to the optic disc size or age. Statistical analysis was done by regression analysis. RESULTS: With an increase in optic disc size, the increase in cup shape, effective area, 1/2 depth area, C/D ratio, neuroretinal rim area, volume above, volume below, and 1/2 depth volume were statistically significant (p < 0.05). However, contour variation, mean contour depth, average depth, maximum depth, average slope, and maximum slope were not affected (p > 0.1). Age did not have any significant influence on optic disc variables (p > 0.1). CONCLUSION: Optic disc size, but not age, should be considered in the interpretation of optic disc variables.

Adult↗

Use of meiotic FISH for identification of a new monosome in Gossypium hirsutum L.

The extensive use of molecular cytogenetics in human genetics and clinical diagnostics indicates that analogous applications in plants are highly feasible. One sort of application would be the identification of new aneuploids, which traditionally involves either direct karyotypic identification, which is feasible in only a few plant species, or tests with markers (cytogenetic, genetic, or molecular), which require sexual hybridization and at least one subsequent seed or plant generation. We have used meiotic fluorescence in situ hybridization (FISH) to analyze a new monosome of cotton (Gossypium hirsutum L., 2n = 4x = 52, 2(AD)1) that had a phenotype which seemed to be distinct from monosomes in the Cotton Cytogenetic Collection. Painting with A2-genome DNA revealed the monosome's D-subgenome origin. DAPI-PI staining showed that the monosome carries a major NOR, delimiting it to the major NOR-bearing chromosomes of the D-subgenome, i.e., 16 or 23. Dual-color FISH with 5S and 18S-28S rDNAs indicated that the monosome contains separate major clusters of each of these two tandemly repeated rDNA elements, thus delimiting the monosome to chromosome 23, for which the Cotton Cytogenetic Collection has previously been devoid of any sort of deficiency. Of the 26 chromosomes in the cotton genome, the Collection now provides coverage for 16 (70%) in the form of monosomy, and 20 (77%) in the form of monosomy and (or) telosomy. Use of molecular cytogenetic methods to identify a new plant aneuploid in cotton exemplifies the fact that a physicochemical karyotypic chromosome identification system is not required a priori for application of new molecular cytogenetic methods, thus indicating their potential applicability to nearly all plant species.

Fluorescent Dyes↗

[The prospective study of the influence of cytomegalovirus (CMV) infection of pregnant women to fetus].

Screening of CMV infection in 450 pregnant women in Shenyang was carried on by ELISA and PCR methods, and CMV infection was detected in 100 of their infants prospectively. The results were 97.11% of pregnant women with past infection, 0.89% with primary infection, 11.11% with recurrent infection, only 2% were susceptible. From 450 pregnant women, there were 3 infants with congenital defects and 3 abortions in active infective group, who were positive for IgM antibody, CMV DNA were positive in heparin peripheral blood, cervical secretion, placenta or amnionic fluid. The fetal infection rate and congenital defect rate in infective group were higher than that in inactive control group. Within 100 maternal-infant pair group, congenitally infected infants were 1.43 times higher in infective group than that in control group (RR = 1.43). Two infants with mental retardations (MDI<70) were born in infective group, but none in control group. In our study, primary infection of early pregnancy was most harmful to fetus, the intrauterine transmission rate was 33.3%. Among nine infected infants born from the infective group, two infants were with CID, and seven were symptomless. CMV screening is important for pregnant women in view of eugenic. When active CMV infection is found in early pregnancy, termination of pregnancy is better considered while CMV detection of amniotic fluid is also positive.

Adult↗

[Studies on peripheral blood stem cells mobilization by short course and high-dose G-CSF].

OBJECTIVE: To explore the effect of short-course, high dose granulocyte-colony stimulating factor (G-CSF) on peripheral blood stem cell (PBSC) mobolization. METHODS: G-CSF 5microg/kg was injected subcutaneously twice daily for 3 consecutive days. On the first day and the fourth day the peripheral blood and bone marrow were collected for CFU-GM, BFU-E, CFU-mix assays and CD34/CD38 detection. A single apheresis of over 8000ml blood processing was performed with CS-3000 blood cell separator. The mononuclear cells were prepared for hematopoietic colony assays and CD34/CD38 detection before and after freezing. RESULTS: After adminitration of G-CSF, the white blood cell count increased significantly. The CFU-GM, BFU-E, CFU-mix yields and CD34+ CD38+ cell and CD34+ CD38- cells also increased significantly (P<0.05). The median of mononuclear cells was 1.56 x 10(8)/kg. After one week freezing, the recovery of mononuclear cells, CFU-GM, BFU-E yields and CD34+ CD38+ cells were about 80% (P>0.05) and CFU-mix yields was 60%; The CD34+ CD38- cells were decreased compared with that of pre-freezing (P<0.05). The median of the reinfused mononuclear cells was 1.27 x 10(8)/kg,CFU-GM 6. 7 x 10(4)/kg, BFU-E 1.6 x 10(4)/kg, CFU-mix 0.32 x 10(4)/kg, CD34- CD38+ cells 1.6 x 10(6)/kg and CD34+ CD38- cells 0.23 x 10(6)/kg. CONCLUSION: Short course and high-dose G-CSF can effectively mobilize the peripheral stem cells. The mononuclear cells, hematopoietic colonies and CD34/CD38 cells were satisfied for the peripheral stem cells rescue.

ADP-ribosyl Cyclase 1↗

Protein engineering on subtilisin E.

Protein engineering was carried out by site-directed and random mutagenesis on subtilisin E gene. Four mutants were obtained. They are M222A; M222A, N118S; M222A, N118S, Q103R; and M222A, N118S, Q103R, D60N. The mutant genes were recombined in pBE-2, an E. coli-B. subtilis shuttle vector, and transformed into B. subtilis DB104, an alkaline and neutral proteinase deficient strain. The subtilisin E mutations obtained from their gene expressions were purified. The properties of these mutants showed that the M222A mutation made the enzyme resistant to oxidation, N118S mutation increased the thermal stability, while Q103R and D60N mutations enhanced the specific activity of the enzyme but decreased the thermal stability and, in particular, D60N mutation caused the enzyme to be very unstable. The IEF-PAGE showed that the wild type and M222A mutant had the same pI of 8.92, while those of double mutant, triple mutant, and quadruple mutant were 8.88, 9.10, and 9.17, respectively. The optimum pH range was 7.5-9.5 for suc-AAPF-pNA substrate and was 10-12 for casein substrate.

Catalysis↗

Familial occurrence of hemangioblastoma of central nervous system.

OBJECTIVE: To study the familial occurrence of hemangioblastoma of the central nervous system and to search for an effective way to diagnose and manage the disease. METHODS: From 1974 to 1993, six patients with hemangioblastoma with family history underwent surgical treatment at our hospital. Computerized tomography (CT) or magnetic resonance imaging (MRI) technique was used for the diagnoses before operation. RESULTS: Each patient had more than two relatives with hemangioblastoma. Multiple lesions were found in two of the six familial hemangioblastoma cases. All the patients recovered well after operation. So far, no victims have been discovered in the third generation. CONCLUSIONS: The high occurrence rate of familial hemangioblastoma cases suggests an involvement of hereditary factors. We need a long follow-up study on the descendants of familial hemangioblastoma patients. MRI is of diagnostic value in determining the location and nature of hemangioblastoma in both brain and spinal cord. A thorough examination is necessary for hemangioblastoma patients, and microsurgery is important in removing the tumor.

Adult↗