Biomedical subjects

Yong Zhao

Publications and source records attributed to Yong Zhao.

At least 73 records · Page 4Linked to original sources

PubMed2004

Achievement of cellular immunity and discordant xenogeneic tolerance in mice by porcine thymus grafts.

Specific cellular immune tolerance may be essential for successful xenotransplantation in humans. Thymectomized (ATX), T and NK cell-depleted immunocompetent mice grafted with xenogeneic fetal pig thymic and liver tissue (FP THY/LIV) result in efficient mouse thymopoiesis and peripheral repopulation of functional mouse CD4(+) T cell. Very importantly, the reconstituted mouse T cells are specifically tolerant to pig donor antigens. Studies demonstrated that porcine MHCs mediated positive and negative selection of mouse thymocytes in FP THY grafts, whereas mouse MHCs were involved in negative selection in grafts. Therefore, T cell tolerance to xenogeneic donor antigens could be induced by grafting donor thymus tissue. Xenogeneic thymic replacement might have a potential role in the reconstitution of cellular immunity in patients with AIDS or other immunodeficiencies caused by thymus dysfunction.

Acquired Immunodeficiency Syndrome↗

PubMed2004

Altered allogeneic immune responses in middle-aged mice.

It is well known that leukocyte composition, T cell phenotypes and immune function change in aged mice and humans. However, limited and conflicting results on the age-related immune changes in middle-aged mice were reported. Identification of the characteristics of allogeneic immune responses in aging mice may offer important information for transplantation immunology. The major age-related changes in the immune cell phenotypes and function of 12 months old mice include: 1) the significantly decreased CD4+ cell population in the peripheral blood, the major peripheral CD4+ cells is CD45RB(low)CD62L(low) memory phenotype; 2) the in vitro responses to alloantigens and Con A of splenocytes markedly reduced; 3) the in vivo secondary humoral immune responses to alloantigens significantly declined; 4) the age-related alteration in the thymus mainly occurred in CD4/CD8 double positive (DP) stage; and 5) increased CD80+ and MHC class II+ cell population in spleens. Thus, the major age-related immune changes in 12 months old mice occurred in CD4+ T cells in the periphery and DP stage in the thymus, which may subsequently lead to the decreased allogeneic immune responses and the different sensitivity to immunosuppressive drugs and treatments. Further studies on the characteristics of allogeneic immunity in aging individuals may help to determine the appropriated treatment for transplant aging individuals.

Aging↗

PubMed2003

Suppression of mitogen-activated protein kinase phosphatase-1 (MKP-1) by heparin in vascular smooth muscle cells.

Heparin inhibits vascular smooth muscle cell (VSMC) proliferation, but mechanisms remain elusive. Because heparin inhibits signaling through multiple kinase cascades, we investigated the possibility that phosphatases could be involved. Mitogen-activated protein kinase phosphatase-1 (MKP-1) was the predominant MKP detected in VSMC lines. MKP-1 protein was increased by serum stimulation of quiescent cells, and this increase was diminished by heparin (1 microg/mL). Increased MKP-1 expression was dependent on the mitogen-activated protein kinase, Erk. Decreased Erk activity in the presence of heparin preceded, and may account for, decreased MKP-1. The antimitogenic effects of heparin are therefore unlikely to act through a shift in the kinase/phosphatase balance, but rather through direct kinase suppression. However, because MKP-1 is known to cause an increase in activity of kinases upstream of Erk, that may signal through additional pathways, the decrease in MKP-1 activity may paradoxically enhance heparin's antiproliferative effects. VSMC selected to grow in the presence of heparin express decreased levels of MKP-1 that are unresponsive to heparin, and Erk activity becomes unresponsive to heparin in one cell line. We conclude that phosphatase activation is not a direct mechanism of suppression of multiple kinase cascades by heparin.

Animals↗

PubMed2003

Low-energy (30 keV) carbon ion induced mutation spectrum in the LacZ alpha gene of M13 mp 18 double-stranded DNA.

Double-stranded M13 mp 18 DNA was irradiated with 30 ke V carbon ions in dry state under vacuum to investigate the low-energy heavy ion induced mutation spectra. When the irradiated DNA was used to transfect Escherichia coli JM 105, 3.6-5.7-fold increases in mutation frequency were observed, in contrast to the spontaneous group. Sequences of the 92 induced mutants showed that the carbon ions in this study could induce an interesting mutation spectrum in the lacZ alpha gene. One-base mutations (96.8%) and base pair substitutions (56.4%) were predominant, most of which involved G:C base pairs (90.6%), especially G:C --> T:A transversions (49.6%) and G:C --> A:T transitions (39.6%). This is similar to the spectra induced by gamma-rays in the same ds M13, wild type E. coli system. We also found a considerable amount of carbon ion induced one-base deletion (38.5%) and the mutation sites distribution on the target lacZ alpha gene was obviously non-random. We compared this study with previous data employing gamma-rays to discuss the possible causes of the mutation spectrum.

Base Sequence↗

PubMed2003

Despite efficient intrathymic negative selection of host-reactive T cells, autoimmune disease may develop in porcine thymus-grafted athymic mice: evidence for failure of regulatory mechanisms suppressing autoimmunity.

BACKGROUND: CD4 T-cell reconstitution and xenogeneic tolerance is achieved in T cell-depleted, thymectomized C57BL/6 (B6) mice and nude mice by grafting of fetal pig thymus (FP THY). Sixty percent of grafted nude mice and 10% of grafted thymectomized B6 mice develop a clinical illness resembling chronic graft-versus-host disease. METHODS: Negative selection of mouse T cells in FP THY grafts was studied in "AND" TCR transgenic mice with a negative selecting MHC. Pathologic and immunohistochemical examinations and adoptive transfer assays were performed to determine the role of mouse CD4+ cells in the occurrence of autoimmune disease in this model. RESULTS: Marked clonal deletion of mouse thymocytes bearing a transgenic TCR ("AND"), which recognizes H2s expressed by host hematopoietic cells, was observed in FP THY grafts. Pathologic and immunohistochemical examinations of the liver, skin, lungs, and kidneys of mice with wasting syndrome showed marked mouse CD4+ T-cell infiltration without detectable pig cells. After adoptive transfer of splenocytes, but not of CD4+ cell-depleted splenocytes, from sick mice along with B6 bone marrow cells to lethally irradiated syngeneic B6 mice, the secondary recipients developed a similar autoimmune syndrome as the donors. Cotransfer of naïve syngeneic splenocytes prevented the occurrence of autoimmune disease in secondary recipients of splenocytes from healthy FP THY-grafted BALB/c nude mice. CONCLUSION: These results demonstrate a key role for mouse CD4+ T cells in causing autoimmune disease in this model and suggest the importance of regulatory mechanisms in addition to intrathymic clonal deletion for the maintenance of tolerance to recipient antigens.

Adoptive Transfer↗

PubMed2003

NK cell tolerance in mixed allogeneic chimeras.

Alterations in inhibitory receptor expression on NK cells have been detected in mixed allogeneic chimeras and in mosaic MHC class I-expressing transgenic mice. However, it is not known whether or not NK cells are tolerant to host and donor Ags in mixed chimeras. In vitro studies have shown a lack of mutual tolerance of separated donor and host NK cells obtained from mixed chimeras. Using BALB/c-->B6 fully MHC-mismatched mixed chimeras, we have now investigated this question in vivo. Neither donor nor host NK cells in mixed chimeras showed evidence for activation, as indicated by expression of B220 and Thy-1.2 on NK cells in chimeric mice at levels similar to those in nonchimeric control mice. Lethally irradiated, established mixed BALB/c--> B6 chimeras rejected a low dose of beta(2)-microglobulin-deficient bone marrow cells (BMC) efficiently but did not reject BALB/c or B6 BMCs. In contrast, similarly conditioned B6 mice rejected both BALB/c and beta(2)-microglobulin-deficient BMCs. Thus, NK cells were specifically tolerant to the donor and the host in mixed allogeneic chimeras. The similar growth of RMA lymphoma cells in both chimeric and control B6 mice further supports the conclusion that donor BALB/c NK cells are tolerant to B6 Ags in chimeras. Administration of a high dose of exogenous IL-2 could not break NK cell tolerance in chimeric mice, suggesting that NK cell tolerance in chimeras is not due to a lack of activating cytokine. No reduction in the level of expression of the activating receptor Ly-49D, recognizing a donor MHC molecule, was detected among recipient NK cells in mixed chimeras. Thus, the present studies demonstrate that NK cells in mixed chimeras are stably tolerant to both donor and host Ags, by mechanisms that are as yet unexplained.

Animals↗

PubMed2003

Cloning and characterization of a novel cardiac-specific kinase that interacts specifically with cardiac troponin I.

Cardiac-restricted genes play important roles in cardiovascular system. In an effort to identify such novel genes we identified a novel cardiac-specific kinase gene TNNI3K localized on 1p31.1 based on bioinformatics analyses. Sequence analysis suggested that TNNI3K is a distant family member of integrin-linked kinase. Northern blot and 76-tissue array analyses showed that TNNI3K is highly expressed in heart, but is undetectable in other tissues. Immunohistochemical analysis predominantly localized TNNI3K in the nucleus of cardiac myocytes. In vitro kinase assay showed that TNNI3K is a functional kinase. The yeast two-hybrid system showed that TNNI3K could directly interact with cardiac troponin I, results that were further confirmed by coimmunoprecipitation in vivo. Our data suggest that TNNI3K is a cardiac-specific kinase and play important roles in cardiac system.

Adult↗

PubMed2003

A priori assessment of the stereoelectronic profile of phosphines and phosphites.

This research has demonstrated the utility of a rigorously calibrated, molecular mechanics/semiempirical quantum mechanical protocol for developing stereoelectronic (Tolman) maps for phosphine ligands. A computational analysis of alkyl and aryl phosphines in common usage suggests that these ligands are quite similar stereoelectronically. A noticeable gap in the Tolman map for common phosphines is observed for large, electron-poor phosphines. Several candidates meeting these criteria were identified, the most promising of which is P(t-C(4)F(9))(3). Phosphines in which the phosphorus participates in a ring, which comprise a very small subset of reported phosphines, have very interesting stereoelectronic properties, particularly those in which the ligating phosphorus is part of a three-membered ring. In terms of steric properties, the symmetric deformation coordinate proposed by Orpen and co-workers on the basis of crystallographic studies is calculated with sufficient accuracy using PM3(tm) to allow good confidence in predictions of novel phosphines. For quantification of the electronic properties of phosphines, we analyzed changes in the CO stretching frequency upon changing the ancillary phosphine ligands.

Journal Article↗

PubMed2003

A human peripheral blood monocyte-derived subset acts as pluripotent stem cells.

We have identified, cultured, characterized, and propagated adult pluripotent stem cells (PSC) from a subset of human peripheral blood monocytes. These cells, which in appearance resemble fibroblasts, expand in the presence of macrophage colony-stimulating factor and display monocytic and hematopoietic stem cell markers including CD14, CD34, and CD45. We have induced these cells to differentiate into mature macrophages by lipopolysaccharide, T lymphocytes by IL-2, epithelial cells by epidermal growth factor, endothelial cells by vascular endothelial cell growth factor, neuronal cells by nerve growth factor, and liver cells by hepatocyte growth factor. The pluripotent nature of individual PSC was further confirmed by a clonal analysis. The ability to store, expand, and differentiate these PSC from autologous peripheral blood should make them valuable candidates for transplantation therapy.

Antigens, CD34↗

PubMed2003

[Clinico-pathologic analysis of 26 cases of leiomyoma of the vagina].

OBJECTIVE: To study the clinicopathological features, the histogenesis, and the correlation between the expressions of estrogen receptor(ER) and epidermal growth factor receptor(EGFR) in leiomyoma of the vagina. METHODS: The data of 26 patients with leiomyoma of the vagina were analyzed retrospectively, and S-P immunohistochemistry was used to detect smooth muscle actin(SMA), S-100 protein, CD34, ER and EGFR in the tumor tissues. RESULTS: All patients with leiomyoma of the vagina in this group were married women, with age ranging from 30 to 52 years (mean, 44.5 years). Leiomyoma of the vagina was slow in growth and occasionally recurred with borderline and malignant transformation. Immunohistochemical staining demonstrated to be strongly positive for SMA, and negative for S-100 protein and CD34 in all cases. Positive expression rates for ER and EGFR were 38.5 (10/26) and 34.6 (9/26) respectively. There was a significant positive correlation between the expressions of ER and EGFR (Fisher exact, P = 0.0023, r = 0.75). Resection of the tumor mass was an effective method to cure the disease. CONCLUSION: Leiomyoma of the vagina is a rare condition, which may be derived from stem cells of mesenchyma of the vagina differentiating towards smooth muscle. The estrogen hormone and EGF may play an important role in the development and growth of leiomyoma of the vagina. Follow-up is needed after operation for the patient with borderline tumor.

Adult↗

PubMed2003

Prolongation of allogeneic skin graft survival by injection of anti-Ly49A monoclonal antibody YE1/48.

Ly49A receptors expressed on NK, NKT, and T cells play inhibitory roles in regulating the immune responses in vivo and in vitro. Whether or not injection of anti-Ly49A monoclonal antibody (mAb) YE1/48 can block allograft rejection has not been evaluated. Balb/c mouse recipients received intraperitoneal injections of YE1/48 mAb (0.5 mg) or control mAb or phosphate-buffered saline on days -1 and 10. On day 0, fully MHC-mismatched allogeneic C57BL/6 (B6) skin grafts were implanted. The skin graft survival and anti-donor humoral responses were observed. Whereas allogeneic B6 skin grafts survived 14 days in isotopy control antibody-treated or nontreated Balb/c mice, injection of YE1/48 mAb significantly prolonged B6 skin graft survival to 19 days (P < 0.0005). Injection of YE1/48 mAb into presensitized Balb/c recipients did not significantly delay B6 skin graft rejection. On the other hand, after depleting recipient NK, NKT, and some cytotoxic T cells by injection of anti-asialo GM1, YE1/48 failed to prolong B6 skin graft survival in Balb/c recipients. The present studies indicate that injection of YE1/48 mAb significantly delays allogeneic skin graft rejection in nonsensitized recipients but not in sensitized recipients. The presence of NK, NKT cells, and some cytotoxic T cells may be essential for YE1/48-mediated immunosuppression in vivo.

Animals↗

PubMed2003

Fetal porcine thymus engraftment, survival and CD4 reconstitution in alphaGal-KO mice is impaired in the presence of high levels of antibodies against alphaGal.

Xenospecific T-cell tolerance can be induced among murine and human T-cells by porcine thymic grafting. However, anti-alpha 1,3-galactosyltranserase (alphaGal) (Galalpha1-3Galbeta1-4GlcNAc-R) natural antibodies (NAbs) pose a major barrier to porcine xenografts in humans. We used alphaGal knockout (KO) and muchain KO mice to explore the effect of natural anti-alphaGal and other xenoantibodies on porcine thymic engraftment and to examine the potential of thymic tissue to tolerize anti-alphaGal antibody-producing cells. Thymectomized [adult thymectomy (ATX)] non-immunized and rabbit red blood cell (RRBC) pre-transplant immunized alphaGal-KO (knockout), wild-type (WT) and mu chain KO B6 mice were treated with 3Gy total body irradiation (TBI), and T and natural killer (NK) cell depleting monoclonal antibodies (mAbs). These conditioned mice were grafted with fetal porcine thymus and liver (FP THY/LIV) tissue under the kidney capsule. Flow cytometric analysis was performed to follow CD4 reconstitution as a measure of FP THY engraftment and function. Only mice with >10% CD4+ peripheral blood lymphocytes (PBL) were considered successfully engrafted. Enzyme-linked immunosorbent assay (ELISA) was used to assess the kinetics of immunoglobulin M (IgM) and IgG anti-alphaGal antibodies. Anti-pig antibodies were monitored by flow cytometry (FCM). FP THY engrafted successfully in most of the immunoglobulin deficient mice (11 out of 12, 92%) and the outcome was similar in WT B6 controls (8 out of 12, 67%). Non-immunized alphaGal-KO mice grafted with FP THY had a similar success rate (7 out of 11) to that observed in non-immunized alphaGal-WT controls (2 out of 4). In contrast, alphaGal-KO mice immunized pre-transplant with RRBC, then grafted with FP THY/LIV, showed a significant reduction in the success of thymic grafting (2 out of 9, 22%) compared with pre-transplant immunized WT controls (4 out of 7; 57%) and non-immunized alphaGal-KO mice (7 out of 11, 64%). Anti-Gal and anti-pig antibody levels were not markedly augmented by porcine thymus grafts in mice with successful thymus grafts. FP THY engraftment is impaired in the presence of high levels of anti-alphaGal xenoantibodies. However, low levels of anti-alphaGal antibodies and other mouse anti-pig NAbs appear not to play a major role in the rejection of FP THY. Although grafting FP THY expressing the alphaGal epitope did not tolerize B cells producing anti-alphaGal antibodies in a T-cell independent manner, it prevented T-cell dependent sensitization by inducing T-cell tolerance to porcine antigens.

Animals↗

PubMed2003

Pre-transplant analysis of accommodation in donor pigs.

Accommodation could lead to xenograft acceptance without the need for severe immune suppression. Generally graft accommodation is appreciated in the sensitized recipient, after transplantation. By inducing accommodation in chimeric donors, however, the risk and cost of inducing accommodation in the recipient would be reduced. An indirect assay of accommodation in the donor pig is needed for screening donors prior to procurement of the xenograft. The resistance of peripheral blood lymphocytes to cytolysis by antibody and complement was assessed in chimeric pigs and compared with control pigs. Peripheral blood lymphocytes (PBL) from chimeric pigs demonstrated a wide range of cytolysis (0 to 85%, median 13%) whereas PBL from control pigs were consistently lysed with these conditions (86 to 99%, median 96.5%, P < 0.0001). Accommodation or reduction in cytolysis did not correlate with the amount of chimerism. A longitudinal study demonstrated persistent accommodation of the PBL for as long as 15 weeks, when the donors averaged 68 kg in weight. Accommodation has been induced by low levels of antibodies interacting with the target tissue. An ELISA for sheep IgG was developed and the serum from newborn pigs assessed. Sheep IgG (up to 4.6 microg/ml) was detected in four of seven piglets with chimerism detectable by flow cytometry and in one of four piglets with minimal chimerism, detectable only by PCR. Lymphocyte accommodation was observed in all pigs with detectable sheep IgG. Of four pigs without accommodation, none had sheep IgG. Three pigs without detectable sheep IgG also had accommodation, suggesting that factors other than sheep IgG may induce accommodation. Acute vascular rejection was not observed in the heterotopic heart transplants from six donors with PBL accommodation. Only one incident of moderate diffuse cellular rejection (grade 3) was observed.

Animals↗

PubMed2003

Xenogeneic skin graft rejection in M-CSF/macrophage deficient osteopetrotic mice.

BACKGROUND: The cellular infiltrate in xenografts suggests that macrophages may be involved in xenograft rejection. However, the precise role of macrophages in xenograft rejection has not yet been fully addressed. METHODS: Xenogeneic rat skin grafts were transplanted to macrophage colony stimulating factor (M-CSF)/macrophage-deficient osteopetrotic ([OP]-/-) and wild-type control mice. Skin graft survival and antidonor rat humoral responses were quantified. RESULTS: Xenogeneic rat skin grafts survived 13 days in wild-type control mice, survival of rat skin grafts was significantly prolonged to 24 days in [OP]-/- mice (P<0.01). Similar results were observed in sensitized [OP]-/- and control mouse recipients, showing markedly prolonged rat skin graft survival in [OP]-/- mice. Levels of T-cell-dependent antirat antibodies [immunoglobulin G (IgG)2a and IgG3] in sera of [OP]-/- mice were significantly lower than that of control mice 2 weeks post-rat skin grafting. The proliferative responses to xenogeneic rats not to allogeneic mouse stimulation of T cells from [OP]-/- mice were significantly lower than that of wild-type mice. However, neutrilization of M-CSF by anti-M-CSF monoclonal antibody (mAb) or the addition of M-CSF to the in vitro culture systems of wild-type or [OP]-/- mouse T-responder cells, respectively, did not significantly change proliferative responses and cytolytic function against xenogeneic rat targets of wild-type or [OP]-/- mouse T-responder cells. CONCLUSIONS: The in vitro data indicate that M-CSF does not directly regulate cellular immune responses to xenoantigens. The present studies indicate that macrophages may play an important role in immune rejection of xenografts. The precise role of macrophages in xenograft rejection should be further investigated.

Animals↗

PubMed2003

Numerical simulation of opening process in a bileaflet mechanical heart valve under pulsatile flow condition.

BACKGROUND AND AIM OF THE STUDY: Most previous computational fluid dynamics (CFD) studies of blood flow in mechanical heart valves (MHVs) have not efficiently addressed the important features of moving leaflet and blood-leaflet interaction. Herein, computationally efficient approaches were developed to study these features and to obtain better insight into the pulsatile flow field in bileaflet MHVs. METHODS: A simple and effective method to track the moving boundary was proposed, and an efficient method for calculating the blood-leaflet interaction applied. In this way, a CFD code was developed to study the pulsatile flow field around bileaflet MHVs. The CFD code was parallelized on a supercomputer to reduce turn-around time in the simulation. The solver was then used to study the opening process in a St. Jude Medical (SJM) size 29 bileaflet MHV. RESULTS: CFD results showed that, in the opening process, the flow field was consistently partitioned into two side channels and a central channel due to the presence of the two leaflets. In the flow field near the surface of the two leaflets, the fluid velocity followed the local surface velocity of the leaflets, thus showing a strong blood-leaflet interaction effect. Throughout the valve-opening process, peak velocities were always observed near the tips of the valve leaflet. The CFD simulation showed that the opening process took approximately 0.044 s, which compared well with experimental findings. CONCLUSION: The new computational approaches were efficient and able to address the moving leaflet and blood-leaflet interaction. The flow field in the opening process of a SJM 29 bileaflet MHV was successfully simulated using the developed solver.

Blood Flow Velocity↗

PubMed2003

[The high-sensitivity determination of protein concentration by the enhancement of resonance light scattering of chlorophenol red].

A new resonance light scattering (RLS) assay is presented in this paper. At the optimum pH = 4.0, the weak RLS of chlorophenol red can be greatly enhanced by the addition of proteins due to the interaction between protein and chlorophenol red. A new quantitative determination method for proteins has been developed. The linear range is 0-1.0 mg.L-1 for BSA, and 0-0.75 mg.L-1 for HSA with detection limit of 20.3 micrograms.L-1 for BSA, and 20.4 micrograms.L-1 for HSA, respectively. Besides high sensitivity, the method is characterized by good reproducibility, rapidity of reaction, good stability and few inferring substances. The determination results of the proteins in human serum samples by this method are very close to those obtained using biuret spectrophotometric method, with relative stand deviation of 1.17%-2.42%.

Animals↗

PubMed2003

[Cloning and characterization of a novel cardiac-specific kinase gene p93 related to sarcomere].

Myocyte's contraction is regulated by signal transduction pathway composed with many protein factors, but the definite mechanism is still uncertain. A novel cardiac-specific kinase gene which participates in regulation of signal transduction, p93 named, was cloned from adult heart cDNA library. p93, coding a family member of MAPKKK, localized on 1p31.1 based on bioinformatics analyses. Northern blot and 76-tissue array analyses determined that p93 was merely expressed in heart, but was undetectable in other tissues. Immunohistochemical study showed that p93 predominantly localized in the nucleus of cardiac myocytes. In vitro kinase assay indicated that p93 was a functional kinase capable of autophosphorylation. p93 could directly interact with cardiac troponin I by yeast two-hybrid system assessed utilizing bait plasmid containing p93 C-terminus (733 835 aa) and results were further confirmed by co-immunoprecipitation in vivo. Our data suggest that p93 is a cardiac-specific kinase and may play important role in regulation of sarcomeric contraction protein with signal transduction pathway similar ILK.

Amino Acid Sequence↗

PubMed2003

[Clinical study of effect of tianzhi granule on senile vascular demetia].

OBJECTIVE: To investigate the clinical effect of Tianzhi Granule (TZK) on senile vascular dementia (VaD), which is classified as sthenia of liver-yang. METHOD: Two hundred VaD patients were treated with TZK (0.5 g/bag), which was taken one bag each, three times a day. The treatment course was one month and they were treated for rwo courses. RESULT: TZK could remarkably increase gnosia and activity, with no striking difference from that of positive control group (P > 0.05). Simultaneously, TZK could significantly improve the clinical syndrome of traditional Chinese medicine and viability. It could also drastically reduce the whole blood and plasma viscosity and improve erythrodegeneration and abnormality of aggregation index in the abnormal blood viscosity patients. CONCLUSION: TMC has certain effects on senile VaD.

Aged↗