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Biomedical subjects

S Jiang

Publications and source records attributed to S Jiang.

At least 343 records · Page 19Linked to original sources

Inhibition of HIV-1 infection by a fusion domain binding peptide from the HIV-1 envelope glycoprotein GP41.

A synthetic peptide, corresponding to the sequence (637-666) of the HIV-1 glycoprotein gp41, inhibited the replication of an array of HIV-1 strains. The peptide (637-666) selectively binds to the fusion domain at the N-terminus of gp41, suggesting that inhibition of HIV-1 infection is caused by blocking fusion of HIV-1 with cells or of infected cells with uninfected cells. Since this peptide has antiviral activity against both homologous and heterologous HIV-1 isolates and has no detectable cytotoxicity, it offers a novel approach to chemotherapy and prophylaxis of AIDS.

Amino Acid Sequence↗

Synergistic effect of heme and IL-1 on hematopoietic stromal regeneration after radiation.

Results from this study show that a combination of heme and interleukin-1 (IL-1) treatment resulted in the most improved recovery of hematopoietic-stromal regeneration after sublethal irradiation. Less pronounced effects were obtained when heme or IL-1 were given singly. Sublethal irradiation of mice produced an initial (as early as day 1) intense depression of the hematopoietic system as evidenced by leukopenia. In vivo treatment of animals with heme in combination with IL-1, accelerated hematopoietic and stromal regeneration as determined by hematopoietic spleen colony forming unit assay (CFU-S), erythroid (BFU-E), myeloid (CFU-GM) clonal cultures, long-term bone marrow cultures (LTBMC), and the ability to regenerate hematopoiesis by ectopic (renal) stromal hemopoietic transplantation. Sixteen days after irradiation, leukocyte levels in heme and IL-1 treatment groups were higher than non-treated animals and were near normal values by 27 days. One day after irradiation, the capacity of stromal progenitors to form new bone and hematopoietic cells (ectopic foci) was severely impaired, but recovered after 2-4 weeks. This recovery process was accelerated in heme and IL-1-treated animals. BFU-E, CFU-GM, and CFU-S capacity was also severely impaired in all animals 1-27 days after irradiation. CFU-S was only 0.15% of control by day 1 and 5% of control by day 16. Treatment with heme or IL-1 improved recovery by as much as 70% after 27 days of irradiation. A similar but enhanced recovery was seen for BFU-E and CFU-GM, with erythroid recovery the best. Total cellularity, adherent cell layer (ACL) formation, and clonogenic capacity by LTBMCs (10 weeks) derived from irradiated animals was severely reduced, whereas the hematopoietic capacity by LTBMCs derived from heme- and IL-1-treated animals had recovery values similar to non-irradiated controls. These results suggest therapeutic use of heme and IL-1 after chemotherapy or bone marrow depression may be beneficial.

Animals↗

Etiological investigation of acute post-transfusion non-A, non-B hepatitis in China.

The etiology of acute post-transfusion hepatitis (PTH) non-A, non-B (NANB) in China was investigated with the combination of advanced techniques, including a second generation of enzyme immunoassay for detection of the antibodies to hepatitis C virus (anti-HCV), a reverse transcription and nested polymerase chain reaction (RT-nPCR) for HCV RNA, and a PCR for hepatitis B virus DNA. Of the 57 patients who were diagnosed as acute PTH-NANB, 46 (80.7%) were positive for anti-HCV and 41 (71.9%) had HCV RNA. Combining together, 53 (93.0%) were seropositive for anti-HCV and/or HCV RNA. Surprisingly, 18 of these with HCV markers were also positive for HBV DNA, although they were negative for HBsAg, suggesting that a portion of the patients with acute PTH NANB were coinfected by both HCV and HBV. In addition, 4 (7%) of the patients with acute PTH-NANB had no detectable HCV and HBV markers with the use of the above-mentioned techniques. These results indicate that etiologic agents(s) other than HCV and HBV may also cause acute PTH-NANB or that the current techniques may still not be sensitive enough to detect trace levels of HCV and HBV markers.

Adolescent↗

Bovine oocyte development following different oocyte maturation and sperm capacitation procedures.

Various procedures have been reported for successful in vitro maturation and in vitro fertilization (IVM/IVF) of bovine follicular oocytes. Direct comparisons of these different recommended procedures have been rare. In this research, involving a total of 5,128 oocytes, a series of experiments were conducted to compare oocyte maturation, fertilization, and development in vitro with 2 maturation systems (with or without added hormones) and 3 types of sperm treatment procedures. Oocytes were collected from ovarian antral follicles (2-7 mm in diameter) within 3 hr after slaughter of cows or heifers. Those with intact or at least 4 layers of cumulus cells were selected for IVM/IVF. Oocytes were incubated for 22 hr in either Medium 199 with 7.5% fetal calf serum (M199 + FCS) alone or M199 + FCS with added hormones (M199 + FCS + H; oFSH 0.5 micrograms/ml, oLH 5.0 micrograms/ml, and E2 1.0 micrograms/ml) at 39 degrees C in 5% CO2 and 95% air. For IVF, frozen-thawed sperm were treated with either 0.1 microM calcium ionophore A23187 (A23187) for 1 min, or 10 or 100 micrograms/ml heparin (H10 or H100) for 15 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nuclear transfer in cattle: effect of nuclear donor cells, cytoplast age, co-culture, and embryo transfer.

There are many factors affecting the efficiency of nuclear transfer technology. Some are evaluated here using our novel approach by enucleating oocytes at 20-22 hr after in vitro maturation (IVM), culturing the enucleated oocytes (cytoplasts) for 8-10 hr or 18-20 hr to gain activation competence and then conducting nuclear transfer. In the first experiment, we demonstrated that cumulus cell (CC) monolayer can support some cloned embryos to develop into morulae or blastocysts. Co-culture with CC and bovine oviduct epithelial cell (BOEC) monolayers resulted in no differences (P > 0.05) in supporting the development of cloned embryos (Experiment 2). When in vitro matured oocytes were enucleated at 22 hr after IVM followed by nuclear transfer 18-20 hr later, cleavage and morula or blastocyst development of the cloned embryos were similar to those resulting from the enucleated oocytes which had been matured in vivo (Experiment 3). Frozen embryos as nuclear donor cells worked equally well as fresh embryos for cloning in embryo development which was superior to IVF embryos (Experiment 4). However, fresh embryos resulted in a higher proportion (P < 0.05) of blastomere recovery than did frozen of IVF embryos. Finally, embryo transfer of cloned embryos from our procedure produced a viable calf, demonstrating the commercial value of this novel approach of the technology.

Animals↗

The hematopoietic stromal microenvironment promotes retrovirus-mediated gene transfer into hematopoietic stem cells.

In this study we report on the establishment of novel conditions which permit efficient retrovirus-mediated gene transfer of human adenosine deaminase (ADA) into murine hematopoietic progenitors. Using Southern blot analysis and an ADA probe, we demonstrated that prestimulation of bone marrow cells over an in vitro culture of adherent stromal cell layers (ACLs) for two days provides favorable conditions for gene transfer in the absence of exogenous growth factors. In bone marrow transplant recipients reconstituted with retrovirally-marked cells, ADA was detected in spleen, thymus and bone marrow cells of the recipients eight months after transplantation. These observations were also seen in transplants of embryonal hematopoietic stem cells. By using different incubation protocols, it was found that the developmental fate of hematopoietic stem cells varied with the presence of exogenous growth factors or an ACL in the prestimulation phase. Polyclonal hematopoiesis with multiple clones appearing simultaneously was revealed in mice reconstituted with growth factor-stimulated cells four months after transplantation. This was detected by multiple integration patterns of ADA integration into the genomes of individual colony forming units-spleen (CFU-S) in transplantation recipient mice. In contrast, two to five months after transplantation, polyclonal hematopoiesis was not observed in mice reconstituted with cells infected in the absence of growth factors. It appears that utilization of the bone marrow microenvironment through the use of an ACL results in a narrower spectrum of integration patterns, suggesting that a type of oligoclonal or monoclonal hematopoiesis is occurring. These studies demonstrate that an ACL provides novel conditions for successful gene transfer and stable integration of the vector into the genome. Use of an ACL may be advantageous for successful hematopoietic stem cell gene therapy.

Adenosine Deaminase↗

Improbability of harmful autoimmune responses resulting from immunization with HIV-1 envelope glycoproteins.

Autoimmunity mediated by cross-reactive antibodies, elicited by HIV-1 envelope glycoproteins gp120/gp160, has been postulated to contribute to the pathogenesis of AIDS. Partial amino acid sequence homology between gp120/gp160 and several human host proteins, including MHC antigens and immunoglobulins, has been perceived as the basis for immunological cross-reactivity. Binding of antibodies from sera of HIV-1-infected individuals to selected host proteins and/or to synthetic peptides derived from them and the inhibitory activity of such sera in assays measuring the functional activity of T cells provided apparent support for the autoimmunity hypothesis, which is also relevant to the issue of safety of anti-HIV-1 vaccines. Considering the possibility that the detected autoantibodies may arise for reasons other than antibody responses to gp120/gp160, the immunological cross-reactivity between gp120/gp160 and the relevant host proteins was investigated using hyperimmune rabbit anti-gp120/gp160 and monoclonal antibodies. As determined from dilution end-point comparisons for polyclonal anti-gp120, the cross-reactivity of anti-gp120 with CD4 was undetectable (< 10(-5)%). The cross-reactivity of anti-gp120/gp160 with HLA-I and HLA-II antigens was also undetectable (< 4 x 10(-4)%) and that with other human proteins reported to have partial sequence homology with gp120/gp41 was < or = 0.013%. Anti-gp120/gp160 did not have detectable inhibitory effects in functional assays measuring proliferative T cell responses. Therefore, immunization with gp120/gp160 is unlikely to elicit harmful autoimmune responses.

Amino Acid Sequence↗

Characterization of adhesive interactions between human endothelial cells and megakaryocytes.

Cell-cell adhesion is essential for many immunological functions and is believed to be important in the regulation of hematopoiesis. Adhesive interactions between human endothelial cells and megakaryocytes were characterized in vitro using the CMK megakaryocytic cell line as well as marrow megakaryocytes. Although there was no adhesion between unactivated human umbilical vein endothelial cells (HUVEC) and megakaryocytes, treatment of HUVEC with inflammatory cytokines such as IL-1 beta, tumor necrosis factor alpha, INF-gamma, or the phorbol ester phorbol myristate acetate (PMA) resulted in a time- and dose-dependent increase in adhesion. Stimulation of marrow megakaryocytes or CMK cells with the cytokines IL-1 beta, GM-CSF, IL-6, IL-3, or PMA augmented their adhesion to endothelium. Monoclonal antibodies against the LFA-1 subunit of the leukocyte adherence complex CD18 inhibited the binding of marrow megakaryocytes or CMK cells to HUVEC. Adhesion blocking experiments also demonstrated that the VLA-4/VCAM-1 pathway was important for megakaryocyte attachment to HUVEC. Adhesion promoted maturation of megakaryocytic cells as measured by increased expression of glycoproteins GpIb and GpIIb/IIIa and by increased DNA content. These observations suggest that alterations in megakaryocyte adhesion may occur during inflammatory conditions, mediated by certain cytokines, resulting in augmented megakaryocyte maturation.

Bone Marrow↗

Long-term bone marrow stromal and hemopoietic toxicity to AZT: protective role of heme and IL-1.

We studied the immediate and long-term effects of azidothymidine (AZT) and heme on murine hemopoietic and stromal progenitor cells in vivo and in vitro. Treatment of mice for 37 days with AZT produced anemia and leukopenia, whereas combined treatment with heme abrogated some of the toxic effects which were apparent even 2 weeks after cessation of treatment. Quantitation of spleen (CFU-S), erythroid (BFU-E) and myeloid (CFU-GM) colony formation from AZT-exposed animals revealed reductions in these progenitors, and this was partially reversed after heme treatment, especially when mice were allowed a 2-week recovery period. Long-term bone marrow cultures (LTBMC) of cells from treated groups revealed difficulty in establishing an adherent cell layer (ACL) by the first week in culture. Total cellularity, CFU-S, BFU-E and CFU-GM clonogenic potential of cultures remained depressed throughout 10 weeks of culture, whereas heme treatment overcame these depressions when AZT-exposed mice were allowed to recover for 14 days prior to culture of their cells in LTBMC. Interleukin-1 (IL-1) treatment to the same recovery group of AZT-exposed mice also resulted in an improvement of CFU-GM growth in LTBMC that was not seen in the nonrecovered group. Transplantation of cells from treated mice under the renal capsule of recipient mice revealed that AZT depressed the regeneration of osteogenic and hemopoietic cell growth within ectopic foci. These effects were reversed with heme treatment in vivo. In other experiments, heme was found to inhibit human immunodeficiency virus (HIV-1) reverse transcriptase and to potentiate the activity of AZT triphosphate against HIV-1 reverse transcriptase. In summary, these results demonstrate that AZT inhibits the growth and development of a variety of hemopoietic, stromal and adherent cells in vivo and in vitro. Treatment of animals with heme produced recovery to near normal levels and suggests possible therapeutic potential.

Anemia↗

Percutaneous transseptal balloon valvuloplasty for dilating mitral valve stenosis (report of 200 cases).

Two hundred cases with mitral valve stenosis were treated by percutaneous single balloon (Inoue balloon) valvuloplasty at our institute from May 1988 to July 1992. The subjects included 137 females and 63 males with a mean age of 36.5 +/- 8.8 years (15-58 years). Hemodynamic and left ventriculographic findings were evaluated immediately before and after the procedure, and the results showed that the mean left atrial pressure was reduced from 25.08 +/- 9.13 mmHg to 10.64 +/- 4.10 mmHg (P < 0.001), the pressure gradient across the mitral valve was reduced from 25.49 +/- 10.22 mmHg to 6.71 +/- 4.87 mmHg (P < 0.001), and systolic pulmonary pressure was reduced from 52.78 +/- 21.42 mmHg to 38.56 +/- 16.47 mmHg (P < 0.001). At the same time, cardiac output and mitral orifice area were increased from 3.84 +/- 0.11 L/min to 4.66 +/- 0.28 L/min (P < 0.001) and from 1.08 +/- 0.28 cm2 to 2.20 +/- 0.47 cm2 (P < 0.001) respectively. Follow-up of 6-48 months (median 24 months) in 50 nonselective patients showed a symptomatic improvement rate of 100%. Percutaneous transseptal balloon mitral valvuloplasty (PBMV) proved to be a highly effective and safe nonsurgical method that can yield very good results in relieving symptomatic rheumatic mitral stenosis, with minimal morbidity and no mortality in this group. PBMV works by splitting the adhered mitral commissures toward the mitral annulus.

Adolescent↗

A comparison between in vitro fertilization and microinjection of immobilized spermatozoa from bulls producing spermatozoa with defects.

The objectives of this study were to compare the fertilization rate of bovine in vitro matured oocytes by in vitro fertilization (IVF) and by microinjection of a single spermatozoon (MI) and to relate these rates with fertility reported for these bulls in artificial breeding. Bull A (Holstein) had a nonreturn rate of 75%. Semen from this bull is routinely used in our standard IVF procedure. Bull B (Ayrshire), used regularly in artificial breeding and related to bull D, had a nonreturn rate of 69.2%. Bull C (Brown Swiss), with a chromosomal translocation and trisomy, achieved a nonreturn rate of 42%. Bull D (Ayrshire) produced nonmotile spermatozoa (SPZ) and had an abnormality described as "tail stump defect." No pregnancies sired by bull D have been reported. Oocytes were either fertilized in vitro by capacitated SPZ or by microinjection of a single immobilized SPZ into the ooplasm. SPZ were treated with 0.1 microM A23187 and used for IVF. For microinjection SPZ were cocultured for 5 h with bovine oviduct epithelial cells (BOEC) and then immobilized by freezing and thawing twice without cryoprotectant. A single batch of killed SPZ (stored at -25 degrees C) was used for all microinjections. All oocytes were cultured in Medium 199 for 22 h at 39 degrees C and subsequently fixed, stained, and examined for evidence of fertilization (i.e., female and male pronucleus formation, SPZ decondensation). Fertilization rates following IVF with semen from bulls A, B, C, and D were 80%, 54%, 1%, and 2%, and following microinjection were 39%, 22%, 21%, and 34%, respectively.

Animals↗

Synthetic peptides and anti-peptide antibodies as probes to study interdomain interactions involved in virus assembly: the envelope of the human immunodeficiency virus (HIV-1).

Synthetic peptides and anti-peptide antibodies have been widely used as probes to map B- and T-cell epitopes on proteins. Such probes also have the potential to delineate contact sites involved generally in protein-protein interactions or in association of domains within a protein. We applied peptide/anti-peptide probes to define: (1) regions on the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins gp120 and gp41 involved in the association between these two glycoproteins; and (2) sites on gp120/gp41, essential for the association of HIV-1 with the CD4 cell receptor. Results of this examination suggested the following: (1) two segments on gp120, encompassing residues (102-126) and (425-452), contribute to the binding site for CD4 and are expected to be juxtaposed in the folded gp120 chain; (2) portions of immunodominant gp120 and gp41 epitopes, encompassing residues (303-338) and (579-611), respectively, appeared to be involved in the gp120-gp41 association, as suggested by direct binding studies and by the limited accessibility of these epitopes on HIV-1 virions: other portions of gp120 also appeared to contribute to the association between these two glycoproteins; (3) there is a partial overlap between gp41 and CD4 binding sites on gp120; (4) the fusion domain and a segment (637-666) of gp41 are not accessible to antibodies after oligomerization of gp41; and 5) the gp120-gp41 association was blocked by aurintricarboxylic acid, suggesting the possibility of developing antiviral compounds interfering with HIV-1 assembly.

Amino Acid Sequence↗

Fertilization of bovine oocytes after microsurgical injection of spermatozoa.

The objective of this study was to compare the fertilization rate of bovine oocytes matured in vitro (22, 25 or 28 hours) and in vivo (30 to 35 hours after standing estrus) following the microinjection of a single spermatozoon. A single motile spermatozoon was injected into the perivitelline space (Experiments 1 to 9), and a single immotile spermatozoon was injected into the ooplasm (Experiments 10 to 15). A single ejaculate of frozen-thawed semen was used throughout. The spermatozoa were injected either without treatment or after treatment with heparin (100 microg/ml), or Ca ionophore A23187 (0.1 microM), or co-cultured for 5 hours with bovine oviduct epithelial cells (BOEC), or they were co-cultured for 5 hours with BOEC and immobilized by freezing and thawing twice without cryoprotectant, or they remained untreated. Oocytes were placed in a droplet of hyperosmotic solution of 0.1 M sucrose in PBS to enlarge the perivitelline space (Experiments 1 to 9) or in PBS (Experiments 10 to 15). Small amounts of polyvinyl pyrrolidone (PVP) without spermatozoa were injected as a control for parthenogenetic activation. After injection, oocytes were incubated in Medium 199 for 22 hours at 39 degrees C, and they were stained with 1% aceto-orcein and examined for evidence of fertilization or parthenogenetic activation. Low rates (9 to 11%) of fertilization resulted from injection into the perivitelline space of oocytes matured for 22 hours in vitro irrespective of spermatozoa treatment. Fertilization rates were higher in oocytes matured in vivo after injection into either perivitelline space (66%) or ooplasm (74%) than in oocytes matured in vitro (9 to 44% fertilization). Surprisingly, in oocytes matured in vivo, there was no difference in the proportions fertilized by spermatozoa injection into ooplasm and parthenogenetically activated by injection of medium alone (74 and 66%, respectively).

Journal Article↗

Nuclear totipotency of cultured rabbit morulae to support full-term development following nuclear transfer.

The rabbit was used as a model for nuclear transfer. A critical step in nuclear transfer is oocyte activation, which was evaluated in this research. Optimal field strength of an electric stimulus for activation was examined. A significantly higher activation rate in all criteria tested was achieved when oocytes were activated electrically with a field strength of 2.4 kV/cm versus 1.2 or 1.8 kV/cm. Also, electrical stimulation with combined alternating current (AC) and direct current (DC) was superior to DC stimulation alone for activation. In another study involving 586 oocytes, exposure of oocytes to cytochalasin B for 1 h followed by activation with electrical stimulation significantly improved development of the oocytes to blastocyst stage compared to oocytes without cytochalasin B pre-exposure (38% vs 26%, p less than 0.05). Cytochalasin B exposure alone (control), however, had no effect on activation. Exposing oocytes to activation medium without electrical stimulation also activated some oocytes. In the nuclear transfer experiment, blastomeres from 8-cell embryos cultured for 20-24 h to the 32-64-cell stage were used as nuclear donor cells. Of 491 oocytes used, 459 (93%) survived the enucleation and fusion procedure, 370 (81%) fused, and 284 (77%) developed into 2-4-cell embryos. A total of 243 of these 2-4-cell embryos were transferred to 15 pseudopregnant recipients and produced 8 young (3%). Although the efficiency is low, this study demonstrated that rabbit morulae cultured for 20-24 h to the 32-64-cell stage as nuclear donors for transfer remain totipotent.

Animals↗

Correlation between the prevalence of serum HBV DNA and immunoserologic HBV markers in the subjects with or without hepatitis.

The prevalence of serum HBV DNA, detected by polymerase chain reaction, and that of immunoserologic HBV markers (HBsAg, HBeAg, anti-HBs, and anti-HBc), determined by immunoassays, were compared among three groups of subjects: (A) chronic active hepatitis B patients, (B) chronic asymptomatic HBV carriers, and (C) normal individuals. Except five of the normal individuals, all of the subjects are positive for anti-HBc while some of them were also positive for other immunoserologic HBV markers, such as anti-HBs, HBsAg, and HBeAg. Serum HBV DNA were detected in 81% in group A, 52% in group B, and 20% in group C. In both group A and B, serum HBV DNA were detected in all the subjects with anti-HBc+/HBsAg+/HBeAg+. However, the percentage of seropositive HBV DNA in the subjects with anti-HBc+/HBsAg+ in Group A was much higher than that in Group B. Interestingly, the percent of serum HBV DNA+ in the individuals with anti-HBc+ only was markedly higher than that in the subjects with anti-HBc+/anti-HBs+ in both Group A and C, suggesting that anti-HBs may play a role in the inhibition of HBV replication and clearance of HBV virion from blood. Above serological profiles will provide important information concerning the significance of serum HBV DNA detection in judgement of HBV replication in the individuals with or without HBV infection. Cautions should be taken to clarify those so called normal individuals who have no symptoms of hepatitis B, no HBsAg in the sera and normal transaminase, but have HBV replication in their bodies.

China↗