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Biomedical subjects

R Fei

Publications and source records attributed to R Fei.

14 recordsLinked to original sources

Hematopoietic stem cell mobilization after rat partial orthotopic liver transplantation.

BACKGROUND: On the basis of the recently recognized potential of hematopoietic stem cells (HSC) to give rise to hepatocytes, we investigated the possibility that HSC could be mobilized and home to the injured liver promoting tissue repair after 50% partial orthotopic liver transplantation (PLTx) in the rat. METHODS: Using sex-mismatched (female to male) syngeneic SD rats, we performed 50% PLTx or whole orthotopic liver transplantation (WLTx) versus 50% partial hepatectomy (PHx) and sham operation (O). Elements with stem cell markers were detected in peripheral blood (PB) and in the liver. Liver injury and regeneration were estimated. The sex-determining region for Y chromosome gene (SRY) was used to define cell origin by in situ hybridization in liver sections. RESULTS: Comparison of WLTx and PHx groups showed a lower survival rate (50%), in the PLTx group were (P<.05). Further, the liver injury was more serious and the levels of serum biochemical parameters were higher. Compared with PHx groups, on days 3 and 5 postoperatively, the mitosis index and the expression of PCNA were lower among the PLTx groups. Compared with WLTx and sham operation groups, beta2m-/Thy-1.1+, CD34+ cells in PB in PLTx groups and PHx were increased on day 1 postoperatively and decreased on the following days. Compared with PHx groups, beta2m-/Thy-1.1+, CD34+ cells were higher in PLTx. The CD34-, c-kit-, and Thy-1.1-positive cells detected in portal tract areas peaked during 3 to 5 days postoperatively in PLTx. Few SRY+ cells were detected in PLTx liver grafts. CONCLUSIONS: beta2m-/Thy-1.1+ and CD34+ stem cells mobilized after PLTx and PHx may be related to the reduced-size liver. Few HSC are involved in liver regeneration in PLTx.

Animals↗

Differentiation of hematopoietic stem cells into hepatocytes in liver fibrosis in rats.

It has been reported that hematopoietic stem cells (HSC) can differentiate into hepatocytes in the normal liver and in some pathologic environments. The aim of this study was to investigate whether HSC can differentiate into hepatocytes in cases of established liver fibrosis. Rat liver fibrosis was induced by subcutaneous injection of tetrachloride (CCl4). Thy+ CD3- CD45RA- HSC in bone marrow cells, which had been enriched by fluorescence-activated cell sorting (FACS), were labeled with PKH26-GL, and autologously transplanted into CCl4-treated rats. The expressions of albumin (Alb), cytokeratin 8 (CK8), and alpha-smooth muscle actin (SMA) were determined by immunofluorescence methods. The PKH26-GL labeled Thy+ CD3- CD45RA- HSC expressed the hepatocyte-specific markers Alb and CK8, but did not express alpha-SMA in liver fibrosis. Thy+ CD3- CD45RA- HSC differentiated into hepatocytes, but not into hepatic stellate cells. In conclusion, autologous stem cell transplantation may be helpful to treat hepatic fibrosis.

Animals↗

Hemispheric interaction, metacontrol, and mnemonic processing in split-brain macaques.

These experiments explored the interactions remaining between the cerebral hemispheres in two split-brain macaques. The 'split' was earlier confirmed by showing that one hemisphere was incapable of identifying visual images seen by the other. The critical tests for residual interactions were intermingled with control trials in a continuous recognition task. These tests were of two kinds: 'parallel processing', to determine how simultaneous viewing by both hemispheres affected subsequent recognition by one of them alone; and 'conflict', where opposite responses were demanded from the two hemispheres, thus assessing the issue of metacontrol. Two types of stimuli were also employed: ART, in which each hemisphere saw essentially the same image; and BIPARTITE, in which images were entirely different for each hemisphere. Since, with either type of stimulus, performance was best when viewed by both hemispheres at both encoding and retrieval, 'parallel processing' was highly efficient. However, when both hemispheres viewed initially and only one was subsequently queried, performance was significantly worse than when each hemisphere acted alone on each occasion. It is thus reasoned that when both hemisphere view together, the resultant memory trace somehow reflects the bilaterality, a conclusion concordant with observations of Marcel on blindsight. Processing different images (BIPARTITE) was somewhat more disruptive in this regard than if the same image was viewed by each hemisphere. This was particularly true in the conflict situation, where for one hemisphere the item seen was NEW and for the other it was OLD. A response of 'OLD' was, at first, consistently rewarded. When this well-established protocol was changed, the hemispheres in each animal were gradually able to revise their joint behavior. This, together with the effect of disparate images, and the deficiency evoked when the animals were forced to recognize unilaterally an image first viewed under bilateral conditions, all manifest considerable, and complex, interaction between the hemispheres despite absence of the forebrain commissures. The superior colliculus seems a likely focal point for such interhemispheric effects.

Animals↗

[Erythroid differentiation denucleation factor: a family of erythroid regulators for mammalian erythroid terminal differentiation/tumor suppression and the cloning of their related genes].

The role of regulation of erythroid differentiation denucleation factor(EDDF) on mammalian erythroid differentiation and myeloma cell malignancy as well as cloning of their stage related genes were serially studied. Through a series of cybrid and hybridization experiments between mammalian erythroid cells and erythroleukemia or non-erythroid myeloma cells, we have demonstrated a novel family of erythroid regulators(EDDFs) in the mammalian differentiating erythroblasts which with an active peak occurred concomitantly with marked decreases in DNA, RNA and the nuclear anchoring vimentin-IF, but increased in hemoglobin synthesis in cytoplasm prior to the denucleation process during terminal differentiation. The results of cell fusion experiments verified that the supplement of regulators(EDDFs) was critical to the recovery of the originally lost features of terminal differentiation and the reversion of malignant phenotype of tumor cells. Here we showed that the erythroid regulator family EDDFs were essential regulators for the sequential expression of stage related genes of erythroid terminal differentiation, and for the redifferentiation of tumor cells to express the originally inactive globe genes, repressed the oncogenes, and vimentin-IF system, thus initiated nuclear condensation and denucleation. The EDDF gene family consisted of MEDDF, HEDDF-1, HEDRF-1, HEDRF-2 and HCNBP-1 were cloned. All were novel cDNA sequences that have been searched and registered in GenBank. They expressed varying in a stage specific manner, and acted on corresponding genes of terminal differentiation.

Activins↗

[Effect of low dose gossypol treatment on male sperm nuclear basic protein].

OBJECTIVE: Study on human sperm nuclear basic proteins after low dose of gossypol treatment. METHODS: Twenty-three men with normal fertility were divided into two groups. The experimental group included 15 men which were treated daily with gossypol (15 mg/d for 12 weeks and treated with 10 mg/d for 32 weeks continuously), 8 men were used in the control group. Semen was collected at 1, 4, 8, 12, 16, 24, 36 and 44 weeks after gossypol treatment and 10 weeks after stopping treatment. Total nuclear basic protein (TNBP) was extracted and TH/TP, HP1/(HP2+HP3) ratios were determined by scanning microdensitometry following electrophoresis of TNBP in polyacrylamide gels. RESULTS: The TH/TP ratio of control and the gossypol treated group were 0.17 +/- 0.07 and 0.63 +/- 0.79(P < 0.05); The HP1/(HP2+HP3) ratio of control and gossypol treated group were 1.05 +/- 0.21 and 1.47 +/- 0.18 (P < 0.01). These two parameters were returned to normal after 10 weeks of stopping gossypol treatment. CONCLUSIONS: The interruption of histone-to-protamine replacement reaction (HPRR) and alteration of nuclear basic proteins induced by low dose gossypol treatment might lead to infertility. Changes of HPRR and nuclear basic protein were reversible after stopping gossypol treatment.

Adult↗

Long-term reversal of hemispheric specialization for visual memory in a split-brain macaque.

Accuracy of response and pattern of ocular fixations in three split-brain macaques were used to evaluate performance of each hemisphere in a continuous visual recognition task. The animal indicated by ocular fixation upon response points whether a displayed image or face was 'NEW' or 'OLD'. An inadvertent lesion of cingulate gyrus severely reduced contralateral fixations and impaired performance of the affected hemisphere in one animal, confirming the inferred relation between hemisphere and laterality of fixations. The hemispheres in the other two animals were initially remarkably similar in accuracy with human faces and with images; but the right hemisphere was significantly superior to the left for macaque faces. Parallel to this, in the one animal tested while simultaneously using both eyes/hemispheres, fixations were made primarily on the left half of human and macaque faces (right hemispheric control), whereas for images the ocular fixations were predominantly focused on the right half. However, after further, extensive training the left hemisphere performed with significantly greater accuracy than the right on all material and this shift was accompanied and further corroborated by a reversal of the fixational pattern to favor the right half of faces, as continued to be the case with images. Thus, over the long term both the pattern of ocular fixations and the accuracy of performance demonstrate a migration from right to left hemispheric dominance as familiarity with the task increased. Performance of the initially superior hemisphere actually diminished with this shift, presenting a uniquely puzzling question of hemispheric balance in the absence of the forebrain commissures.

Animals↗

Development of a universal immunoenzyme quantitative assay for detecting amplified products of nucleic acid and its preliminary application in hepatitis C virus.

OBJECTIVE: To develop a universal quantitative immunoenzyme assay (EIA) for detecting amplified products of nucleic acid and its application in hepatitis C virus (HCV). METHODS: The appropriate cycle number of amplification was selected to stop polymerase chain reaction (PCR) before the "plateau stage". At the same time, primers HCV (3) of the second PCR were modified with biotin so that the amplified products were labeled. The products were diluted and subsequently added to the streptavidin-coated wells, and the biotinylated products were captured, followed by denaturation of NaOH, and non-biotinylated strands were removed. Hybridization was performed by adding the specific probe labeled with fluorescein. Finally antifluorescein horse radish peroxidase (HRP) conjugates were added, after washing, 3, 3', 5, 5',-tetramethylbenzidine (TMB) was added to the wells and then measured on a microplate reader. RESULTS: EIA detection of amplified products of HCV showed that this assay was rapid, sensitive, specific and accurate. Correlation between the initial number of viral template and the EIA of amplified products was good. We also prospectively investigated the response to interferon in five patients with HCV coinfection. Results showed that this assay could be used as a guidance to the clinical therapy in directing the use of antiviral drugs. CONCLUSIONS: This assay could be widely used as a universal technique for the quantitative detection of amplified products of all nucleic acid (such as virus, bacterium) and other human genes (such as HLA B27), it has vast vistas.

Hepacivirus↗

Analysis of low-speed rotational atherectomy for the reduction of platelet aggregation.

Platelet activation is an important determinant of acute outcomes of percutaneous intervention. The objective of this study was to assess the effect of rotational atherectomy on platelet activation in an in vitro model. Freshly collected heparinized porcine blood was exposed to a 2.0-mm Rotablator burr rotating at one of three speeds: 180,000, 140,000, or 0 rpm. The specimens were analyzed immediately for concentration and size of platelet aggregates and plasma-free hemoglobin. There were significantly more platelet aggregates of >20-microm diameter at higher speeds (7,434+/-2,193 at 180,000, vs. 2,269+/-627 at 140,000, vs. 633+/-258 aggregates/ml at 0 rpm; P < 0.001). Plasma-free hemoglobin, a simple measure of cell damage, decreased with decreasing rotational speed (429+/-168 mg/dl at 180,000, vs. 88+/-44 mg/dl at 140,000, vs. 9+/-9 mg/dl at 0 rpm; P < 0.0001). In vitro, platelet activation decreases with decreasing burr speed, suggesting that the use of the Rotablator system at its minimum approved speed (140,000 rpm) could prove clinically beneficial.

Animals↗

Experimental hypersensitivity pneumonitis: location of transferring cells.

Cultured cells from Micropolyspora faeni-sensitized donors can adoptively transfer murine experimental hypersensitivity pneumonitis (EHP). We sought to determine the location of transferred cells in recipient animals, the influence of the origin of the cultured cells, and the effect of specific intratracheal challenge. We labeled cultured sensitized spleen or lung-associated lymph node (LALN) cells with CFDA-SE, a cytoplasmic stain, before transfer to naive recipients, which were sacrificed 1 h, 1 day, or 4 days thereafter. We also transferred labeled cultured spleen cells to recipients that were challenged with intratracheal M. faeni and sacrificed 4 days later (MF). Controls were recipients of M. faeni-sensitized and cultured cells challenged with intratracheal normal saline (NS) and recipients of ovalbumin (OVA)-sensitized cells cultured with M. faeni and challenged with intratracheal M. faeni (OVA). The number and proportion of cells that were stained were determined in dispersed spleen, peripheral and lung-associated lymph nodes, and lung parenchyma. The extent of the pulmonary inflammatory response was measured by determining the proportion of microscopic fields that were abnormal and the total number of dispersed pulmonary cells. CFDA-SE stained cells uniformly, and stained cells could be detected in recipients for up to 7 days after transfer. CFDA-SE treatment (0.5 microM) did not affect the ability of cells to transfer EHP adoptively. Transferred cells could be detected easily in lung, lung-associated and peripheral lymph nodes, blood, and spleen. Transferred cells localized to the lung at 1 h but then rapidly decreased with no difference between labeled cells from spleen and LALN. After intratracheal M. faeni challenge, there was no difference in the proportion of labeled cells in the lung among any of the groups (MF, NS, or OVA). There was an increase in the number of lung cells in the MF group compared with the control (NS and OVA) groups. We conclude that cells capable of transfer are transiently (1 h) trapped in the lung but are much decreased in the lung by four days after transfer. After intratracheal antigen challenge of recipients, there is a substantial increase in the number of pulmonary cells in animals exhibiting adoptive EHP but not in the control groups. Transferred cells responsible for EHP are increased in the lungs of animals with adoptive EHP.

Adoptive Transfer↗

Mid-Infrared Diode Laser Spectroscopy of X ; 1 A ' HC35 Cl

The observation of the C-Cl stretching fundamental of HC35 Cl in its ground X ; 1 A ' state is reported. HCCl was formed in a continuous flow system by the excimer laser photolysis of CHBr2 Cl at 193 nm and the transient absorption recorded by infrared diode laser spectroscopy. The band has a -type character and the fundamental vibrational frequency is found to be 811.5963 cm-1 . No evidence was found for perturbations in either the ground or vibrationally excited states, implying that the excited triplet a 3 A " states lies above the C-Cl stretching level in HCCl. The observed rotation-vibration transitions were well fit by a standard asymmetric rotor Hamiltonian. There is a decrease in the A rotational constant on vibrational excitation, suggesting that some bend-C-Cl stretch coupling is present.

Journal Article↗

Experimental hypersensitivity pneumonitis: in vitro effects of interleukin-2 and interferon-gamma.

Cultured CD4+ cells are responsible for transfer of adoptive murine experimental hypersensitivity pneumonitis (EHP) (ARRD 1992; 146:1582-8). To characterize interactions that occur in vitro that result in cells able to transfer EHP, we added either antibody to IFN-gamma, antibody to IL-2, or 30 or 300 micrograms/ml IFN-gamma at the onset of 72-hour culture of C3H/HeJ spleen cells from either M. faeni or ovalbumin (control) sensitized donors with 30 micrograms/ml Micropolyspora faeni. We determined the phenotype of cells after culture and the amount of IL-2 or IFN-gamma in the culture supernatants, transferred cells to naive recipients, challenged the recipients intratracheally with M. faeni, and determined the extent of pulmonary inflammatory changes 4 days thereafter. Substantial amounts of IL-2 and IFN-gamma were detected in supernatants of cultures from M. faeni-sensitized animals, and lesser amounts were detected in culture supernatants from ovalbumin-sensitized donors. Treatment of cultures of M. faeni-sensitized cells with antibody to IL-2 or IFN-gamma blocked or reduced measurable IL-2 or IFN-gamma for the duration of culture. Treatment with IFN-gamma blocked increased levels of IL-2 at 48 and 72 hours of culture. Cultured M. faeni-sensitized cells adoptively transfer EHP. Cells from cultures depleted of either IL-2 or IFN-gamma or supplemented with IFN-gamma could transfer EHP equally well. We conclude that in vitro maturation of cells capable of adoptive EHP is not dependent on soluble IL-2 or IFN-gamma and is not altered by exogenous IFN-gamma.

Alveolitis, Extrinsic Allergic↗

Isolation and ex vivo expansion of CD34+ cells from cord blood using dextran sedimentation and avidin column selection.

Human umbilical cord bloods were fractionated by unit gravity sedimentation in 1% (v/v) dextran, followed by immunoaffinity selection for CD34+ stem and progenitor cells. Dextran sedimentation alone enabled recovery of more than 80% of the nucleated cells present and 90% of the CD34+ cells, as determined by flow cytometry. The addition of an immunoaffinity selection step for CD34+ cells resulted in a 134-fold enrichment for CD34+ cells, with a mean yield of 64 +/- 15%. The resultant CD34+ population contained almost half the CFU-GM activity initially present in the cord bloods and could be expanded ex vivo in liquid culture.

Antigens, CD↗

Influence of pressure- and flow-triggered synchronous intermittent mandatory ventilation on inspiratory muscle work.

OBJECTIVE: To determine the effect of pressure- and flow-triggered synchronous intermittent mandatory ventilation on inspiratory muscle work. DESIGN: Consecutive clinical, prospective, randomized trial. SETTING: Medical intensive care unit (ICU) of a U.S. Veterans Affairs Medical Center. PATIENTS: Eight patients recovering from acute respiratory failure of various etiologies. INTERVENTIONS: Assist control, followed by randomized application of pressure- and flow-triggered synchronous intermittent mandatory ventilation at 60%, 40%, 20% of the assist-control rate, and flow-triggered continuous positive airway pressure. A total of eight settings were maintained for 10 mins each. MEASUREMENTS AND MAIN RESULTS: Total work rate (joules/min), inspiratory muscle work (joules/L), and pressure time-product per breath (cm H2O-sec) were measured. During pressure- or flow-triggered synchronous intermittent mandatory ventilation, total work rate increased as the mandatory rate was decreased. The method of ventilator triggering had a significant effect on the total work rate. With pressure-triggered synchronous intermittent mandatory ventilation, the total work rate at 60% of the assist-control rate was similar to that with assist-control; whereas with flow-triggered synchronous intermittent mandatory ventilation, this result was achieved at 40% of the assist-control rate. At a machine support level of 20%, total work rate with pressure-triggered synchronous intermittent mandatory ventilation was significantly greater than with flow-triggered synchronous intermittent mandatory ventilation. The method of ventilator triggering had no significant effect on the inspiratory muscle work of the mandatory breaths. This finding was in contrast to the effect on inspiratory muscle work of spontaneous breaths. With pressure-triggered synchronous intermittent mandatory ventilation, inspiratory muscle work of the spontaneous breaths was greater than with the flow-triggered at machine support of 40% and 20%. With either pressure- or flow-triggered synchronous intermittent mandatory ventilation, inspiratory muscle work of the mandatory breaths was not significantly different from that of the corresponding spontaneous breaths, except at the lower machine support levels with the pressure-triggered synchronous intermittent mandatory ventilation. Pressure-time product followed a trend similar to that of inspiratory muscle work. CONCLUSIONS: During synchronous intermittent mandatory ventilation, the method of ventilator triggering has a significant effect on the total work rate and inspiratory muscle work of the spontaneous breaths, particularly at lower machine support levels. Conversely, the method of ventilator triggering has no significant effect on inspiratory muscle work of the mandatory breaths.

Acute Disease↗

Kinetic studies of haemopoietic progenitor cells (CFU-C) and T-lymphocytes in the liver of human foetus.

In the development of human foetus, liver is an important haemopoietic organ. We have determined the liver weight and total hepatic nucleated cells from 24 foetuses on the one hand, and CFU-C yield, total number of CFU-C and T-lymphocytes in liver from 22 foetuses of different ages on the other. From the kinetic observations through the early developmental period of the foetuses, the maximum yield of CFU-C or the total number of CFU-C in the liver occurs at 4-5 months of gestation; during the first 5 months of gestation the percentage of T-lymphocytes in the liver is about 1-2%.

Cells, Cultured↗