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Biomedical subjects

H Xie

Publications and source records attributed to H Xie.

At least 217 records · Page 12Linked to original sources

Protein kinase C modulates flow-initiated contraction.

The objective of this study was to determine whether protein kinase C (PKC) activation is involved, and what its role might be, in flow-induced contraction. Contractions induced by flow (40 microliters/min), K+ (25 mM) and histamine (300 nM) in segments of the rabbit fatal vein were compared under isometric conditions. The PKC activator 12-O-tetradecanoylphorbol-13-acetate (50 nM) significantly enhanced flow-induced and K(+)-induced, but not histamine-induced, contractions. Down-regulation of PKC by preincubation with phorbol 1,13-dibutyrate (10 nM for 24 hr) markedly inhibited flow contraction. Although histamine-induced contraction was also affected, the response to K+ remained unchanged. The potentiation of flow-induced contraction by 12-O-tetradecanoylphorbol-13-acetate was prevented by the voltage-gated Ca+2 channel antagonist diltiazem (1 microM). When the PKC activator 12-O-tetradecanoylphorbol-13-acetate (50 nM) was added in a Ca(+2)-free medium containing manganese (1 mM), flow-induced and 25 mM K(+)-induced contractions were enhanced and unaltered, respectively, but histamine-induced contraction was reduced. It is argued that of three types of contractions studied, PKC predominantly modulates flow-induced contraction. PKC can influence this contraction through increases in both Ca++ entry and Ca++ sensitivity.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

[Effect of intrahippocampal microinjection of enkephalin on cellular immune function and brain IL-1 alpha gene expression in rat].

The effect of intrahippocampal microinjection of enkephalin on cellular immune function and hippocampal IL-1 alpha gene expression was studied in rats. The results showed that: (1) The proliferation activity of splenic lymphocytes stimulated by Con A was significantly increased with microinjection of Leu-enkephalin 1 microliter (18 mmol/L) or Met-enkephalin 1 microliter (18 mmol/L) into bilateral hippocampus, but was inhibited by Lipopolysaccharide 1 microliter (50 ng/microliter). The proliferative response of splenic lymphocytes to Leu-enkephalin still persisted after bilateral adrenalectomy. Low doses of Leu- or Met-enkephalin (10(-10), 10(-11) mol/L) were also capable of causing proliferative activity of the Con A- stimulated cultured splenic lymphocytes in vitro. (2) Using RT-PCR technique, IL-1 alpha gene expression was detected in hippocampus 90 min after bilateral intrahippocampal injection of LPS. The LPS-induced IL-1 alpha gene expression in hippocampus could no longer be detected 30 min after microinjection of Met- or Leu-enkephalin into hippocampus. The above results suggested that intrahippocampal enkephalin might play an important role in neuro-immunomodulation by enhancing the inhibition of IL-1 alpha gene expression in hippocampal formation.

Adrenalectomy↗

[Evaluation of ligature-induced periodontitis in minipig].

We evaluated the progressing period of ligature induced periodontitis in chinese minipig. Plaque index (PLI), bleeding index (BI), probing pocket depth (PD) and clinical attachment level (CAL) were measured in 3 minipigs. On weeks 4 and 8, there was a significant increase of the mean values of PD and CAL. After this active phase, there was no further destruction up to 20 weeks. Following ligature removed probing depths decreased in 3 weeks, but they did not restore to the baseline. It suggests that using ligature induced periodontitis model to evaluate the treatment therapy and medicine effect have to consider the self-healing of lesions, when a treatment trail is carried out in a model.

Animals↗

Protein kinase C mediates activation of nuclear cAMP response element-binding protein (CREB) in B lymphocytes stimulated through surface Ig.

The cAMP response element-binding protein (CREB) is generally considered to be responsive to elevation of cAMP through the activity of protein kinase A (PKA). Although it is well known that cAMP-raising agents can strongly influence B cell stimulation, the regulation of CREB has been little studied. Recently, cross-linking of surface Ig (sIg) was shown to result in trans-activation of a cAMP response element (CRE)-dependent promoter to which bound B cell CREB. In this study, we explored the mechanism underlying this unexpected linkage between sIg and CREB. We found that sIg cross-linking results in phosphorylation of CREB at Ser133. Although this phosphorylation step is mediated by PKA in pheochromocytoma cells, it depends on protein kinase C (PKC) in B lymphocytes. This conclusion is based on abrogation of sIg-induced CREB Ser133 phosphorylation by long-term phorbol-ester treatment to deplete PKC, and mimicking of sIg-induced CREB phosphorylation and CRE-dependent gene expression by short-term PKC agonism. Furthermore, CD40 ligand (CD40L) and LPS, two PKC-independent forms of B cell stimulation, failed to induce phosphorylation of CREB Ser133. These results suggest that CREB responds to specific surface-receptor signals in B cells and that this response is mediated by PKC. Interestingly, forskolin failed to induce phosphorylation of CREB Ser133 in B cells, although it did so in PC12 pheochromocytoma cells. Taken together with PKC mediation of CREB Ser133 phosphorylation in B cells, these results suggest that the dominant mode of CREB regulation is cell-type specific.

Animals↗

In vitro invasiveness of DU-145 human prostate carcinoma cells is modulated by EGF receptor-mediated signals.

Prostate carcinomas often present an autocrine stimulatory loop in which the transformed cells both express the EGF receptor (EGFR) and produce activating ligands (TGF alpha and EGF forms). Up-regulated EGFR signalling has been correlated with tumor progression in other human neoplasia; however, the cell behaviour which is promoted remains undefined. To determine whether an EGFR-induced response contributes to cell invasiveness, we transduced DU-145 human prostate carcinoma cells with either a full-length (WT) or a mitogenically-active but motility-deficient truncated (c'973) EGFR. The DU-145 Parental and two transgene sublines all produced EGFR and TGF alpha, but the transduced WT and c'973 EGFR underwent autocrine downregulation to a lesser degree, with more receptor remaining intact. DU-145 cells transduced with WT EGFR transmigrated a human amniotic basement membrane matrix (Amgel) to a greater extent than did Parental DU-145 cells (175 +/- 22%). Cells expressing the c'973 EGFR invaded through the Amgel only to about two thirds the extent of the Parental cells (62 +/- 23%). A monoclonal antibody which prevents ligand-induced activation of EGFR decreased the invasiveness of WT-expressing cells by half and Parental cells by a fifth, but had little effect on the invasiveness of c'973-expressing cells; with the result that in the presence of antibody, all three cell lines transmigrated the Amgel to the same extent. The different levels of invasiveness between the three sublines were independent of cell proliferation. These findings demonstrated that EGFR-mediated signals increase tumor cell invasiveness and suggested that domains in the carboxy-terminus are required to signal invasiveness. As an initial investigation into the mechanisms underlying the EGFR-mediated enhanced invasiveness, we determined whether these cells presented different collagenolytic activity, as the major constituents of Amgel are collagen types I and IV. All three sublines secreted easily detectable levels of gelatin-directed proteases and TIMP-1, with WT cells secreting equivalent or lower levels of proteases. The proteolytic balance in these cells did not correlate with invasiveness. These data suggest that the TGF alpha-EGFR autocrine loop promotes invasiveness and that this is accomplished by signalling cell properties other than differential secretion of collagenolytic activity.

Carcinoma↗

Phosphatidyl-2',3'-dideoxy-3'-thiacytidine: synthesis and antiviral activity in hepatitis B-and HIV-1-infected cells.

We recently found that phosphatidyl-2',3'-dideoxycytidine (phosphatidyl-ddC) had substantial anti-hepatitis B virus (HBV) activity in vitro compared to 2',3'-dideoxycytidine (ddC) (Hostetler et al. (1994) Antiviral Res. 24, 59-67). Upon administration of liposomal phosphatidyl-ddC to mice, a 40-fold higher drug area under curve was observed in the liver. To evaluate the possibility of using liver-targeted anti-HBV nucleosides to treat woodchuck hepatitis virus, we wanted to find the most potent and selective lipid conjugates. It has been shown that 2',3'-dideoxy-3'-thiacytidine as a racemic mixture of the cis-isomer (cis-(+/-)-BCH-189) has much greater activity against HBV viruses than ddC in vitro. Recently, it was shown that the (-)-beta-L-enantiomer (3TC) is more active and less toxic than the (+)-beta-D-form ((+)-BCH-189). To determine whether phospholipid conjugates of 3TC retain antiviral activity in 2.2.15 cells as demonstrated previously with ddC, we synthesized the 1,2-dipalmitoyl-sn-glycerol-3-phosphate conjugates of (+/-)-BCH-189 and 3TC and assessed their anti-HBV and anti-HIV activities, in vitro. Phosphatidyl-3TC and phosphatidyl-BCH-189 had antiviral activity comparable to the respective free drugs in 2.2.15 cells which chronically produce HBV. In HIV-1-infected human peripheral blood mononuclear cells and HT4-6C cells, phosphatidyl-3TC and phosphatidyl-(+/-)-BCH-189 exhibited significantly lower activity than the corresponding free nucleosides. In view of the documented ability of phosphatidyl-ddC to target drug to the liver, it seems reasonable to expect that phosphatidyl-3TC or phosphatidyl-(+/-)-BCH-189 could be employed to provide greatly enhanced hepatic antiviral activity in HBV infection in vivo.

Antiviral Agents↗

A diffusing sphere which delivers homogeneous laser light for use in photodynamic therapy.

Photodynamic therapy (PDT) exploits the selective uptake of a photosensitizer in tumors and other hyperproliferative target tissues, as well as the ability to direct the treatment light beam to a specific region. Since the photodynamic effect depends on light dose, tissue optical properties and photosensitizer concentration, uniform delivery of light is crucial to attain optimal photodynamic effect. Many commonly used methods for delivering laser light during photodynamic therapy, such as a free fiber or microlens, require fiber and laser adjustments to obtain a highly uniform beam. In this study, we test the ability of a diffusing sphere to improve the uniformity of a light field coming from an argon laser coupled to a free fiber, in which no attempt has been made to optimize beam characteristics. Light fields from the free fiber, a microlens and the diffusing sphere are compared for uniformity via light intensity readings. An in vivo comparison between the sphere and the free fiber is also made in guinea pigs given Photofrin-II. The diffusing sphere decreases problems with shielding, allows quick and easy application of light by simply applying the device over the desired treatment area, and optimizes the desired photodynamic effect by producing ad highly uniform beam of light with no necessity to optimize light delivery by vibrating, looping or re-cleaving fibers.

Animals↗

Risedronate pretreatment does not hamper the anabolic effects of prostaglandin E2 in ovx rats.

Pretreatment of an anti-resorptive agent on the anabolic effects of prostaglandin E2 (PGE2) was studied on the proximal tibia and tibial shaft of ovariectomy (ovx) rats. Two days after ovx, rats were treated with either risedronate (Ris, 5 micrograms/kg twice weekly) or vehicle (V) for 60 days and then switched to 3 or 6 mg/kg/d PGE2 for 21 or 90 days. Bone area of both proximal tibial metaphysis (PTM) and tibial shaft (TX) were measured. Pretreatment with Ris increased the bone mass in PTM but not in TX of ovx rats. In the PTM, PGE2 produced the same percentage of new bone mass in both V- and Ris-pretreated ovx rats. The amount of new bone was almost the same after 3 weeks and 12 weeks of PGE2 treatment. There was no difference in the anabolic effects of 3 and 6 mg PGE2/kg/d in V-pretreated rats; however, the effects in Ris-pretreated groups were greater with 6 mg PGE2/kg/d than with 3 mg PGE2/kg/d. In TX, only the 6mg PGE2/kg/d administration added new bone on endocortical surfaces of both V- or Ris-pretreatment rats which leads to thickening the minimal cortical width, decreasing the marrow cavity and increasing total bone area. Both doses of PGE2 created new trabecular bone in the marrow cavity of tibial shaft in both vehicle- and Ris-pretreated ovx rats. These results suggest that Ris-pretreatment did not hamper the anabolic effects of PGE2 on either PTM or TX in ovx rats.

Aging↗

Observation of changes in peripheral T-lymphocyte subsets by flow cytometry in patients with liver cancer treated with radioimmunotherapy.

An analysis of lymphocyte subsets was performed in 24 patients with surgically verified unresectable hepatocellular carcinoma (HCC). In all cases, the hepatic artery was cannulated and ligated and a single dose of 131I-Hepama-1 monoclonal antibody (MAb) was infused through a hepatic artery catheter. Blood samples were obtained before infusion and 1 month post-infusion. The results indicated that the CD3+ T-cells (pan T-cells) were normal, whereas the CD4+ T-cells (T-helper/inducer cells) decreased and the CD8+ T-cells (T-suppressor/cytotoxic cells) increased significantly in comparison with the control group. So the CD4+:CD8+ ratio declined markedly. One month post-radioimmunotherapy (RIT), the T-cell subsets did not change, but CD4+, CD8+ and the CD4+:CD8+ ratio differed significantly between patients with and without sequential resection. The changes in the T-cell subsets were closely related to tumour capsule status and the human anti-murine antibody (HAMA) response, but no relationship was found between the tumour and administration of a moderate dose of radioimmunoconjugate. Therefore, determination of changes in the T-lymphocyte subsets and the CD4+:CD8+ ratio could be used as clinical and prognostic indicators in patients who have received RIT. Furthermore, RIT did not affect the distribution of T-cell subsets in patients with HCC.

Animals↗

Molecular basis of attenuation of neurovirulence of wild-type Japanese encephalitis virus strain SA14.

To identify the molecular determinants for attenuation of wild-type Japanese encephalitis (JE) virus strain SA14, the RNA genome of wild-type strain SA14 and its attenuated vaccine virus SA14-2-8 were reverse transcribed, amplified by PCR and sequenced. Comparison of the nucleotide sequence of SA14-2-8 vaccine virus with virulent parent SA14 virus and with two other attenuated vaccine viruses derived from SA14 virus (SA14-14-2/PHK and SA14-14-2/PDK) revealed only seven amino acids in the virulent parent SA14 had been substituted in all three attenuated vaccines. Four were in the envelope (E) protein (E-138, E-176, E-315 and E-439), one in non-structural protein 2B (NS2B-63), one in NS3 (NS3-105), and one in NS4B (NS4B-106). The substitutions at E-315 and E-439 arose due to correction of the SA14/CDC sequence published previously by Nitayaphan et al. (Virology 177, 541-552, 1990). The mutations in NS2B and NS3 are in functional domains of the trypsin-like serine protease. Attenuation of SA14 virus may therefore, in part, be due to alterations in viral protease activity, which could affect replication of the virus.

Amino Acid Sequence↗

Induction of insulin resistance by cholinergic blockade with atropine in the cat.

1. Insulin sensitivity was quantified using a modified euglycaemic technique after hepatic cholinergic blockade with atropine and compared with that after surgical denervation. 2. Intraportal administration of atropine produced dose-dependent inhibition of insulin sensitivity in glucose metabolism. ED50 of atropine was 0.99 mg kg-1 (1 mg = 1.5 microM) with maximum inhibition of 40.3 +/- 11.6%. 3. Atropine (3 mg kg-1) reduced insulin sensitivity by a similar amount (33.6 +/- 3.4%) to that produced by hepatic surgical denervation (37.8 +/- 9.8%). Doses greater than 3 mg kg-1 failed to further alter the insulin resistance produced by surgical denervation or atropine (3 mg kg-1) administration, suggesting that activation of hepatic parasympathetic nerves is necessary to fully express the insulin effect. 4. Atropine reduced insulin sensitivity without changes in plasma concentrations of glucagon or insulin. The temporal response to insulin in this euglycaemic study was not changed after atropine administration or after surgical hepatic denervation. 5. It is suggested that hepatic parasympathetic nerves show a synergistic effect with insulin. Disease states that result in hepatic parasympathetic neuropathy would be expected to produce an insulin resistant liver. 6. The modified euglycaemic clamp method for assessing insulin responses was shown to be reproducible up to four times in the same animal and was sufficiently sensitive and quantitative to be able to generate a dose-response curve in each animal for atropine-induced insulin resistance.

Animals↗

Intracellular Ca2+ release in flow-induced contraction of venous smooth muscle.

We designed the present study to determine whether Ca2+ release from intracellular stores contributes to flow-induced contraction. We carried out experiments on segments of rabbit facial vein under isometric conditions. Intraluminal flow by perfusion of physiological salt solution (10 to 80 microL/min) caused contraction in this vessel, which was significantly inhibited by (1) 30-minute pretreatment with 10 mumol/L ryanodine, the sarcoplasmic reticulum Ca2+ channel opener, and (2) 30-minute pretreatment with concomitant application of 20 mmol/L caffeine and 1 mumol/L cyclopiazonic acid in Ca(2+)-free medium to deplete the sarcoplasmic reticulum. In comparison, contraction initiated by 300 nmol/L histamine was significantly attenuated by the same interventions. K+ (25 mmol/L)-induced contraction was unaffected by ryanodine but was reduced after depletion of the sarcoplasmic reticulum. The phospholipase C inhibitor 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate (10 mumol/L) inhibited contractions induced by flow and histamine but not by K+. These findings indicate that Ca2+ release from intracellular stores, presumably via the phosphatidylinositol pathway, contributes to flow- and histamine- but not raised K(+)-induced contractions in this vessel.

Animals↗

[Study on the lung targeting gelatin microspheres of streptomycin sulphate].

This paper is reported the technology of lung targeting gelatin microspheres of streptomycin sulphate (SMS). The microspheres were prepared with natural biodegradable gelatin as the load material and castor oil as the oil phase. The experimental conditions were optimized, the mean volume diameter obtained being 9.7 microns and the mean rate of encapsulation 15.69%. The content, shape and size of the microspheres showed no remarkable change after storage at 37 degrees C RH 75% for 3 months. Activation energy of heat decomposition E = 75.86kJ/mol. In vitro, the SMS release rate was found to accord with Higuchi equation with t1/2 = 8.6h. In vivo (rabbits) the gelatin microspheres were proved to be concentrated in the lung.

Animals↗

Blocking Ca(2+)-dependent synaptic release delays motoneuron differentiation in the rat spinal cord.

Development of motoneuron electrical properties and excitability was studied in spinal cord explants of rat embryos cultured for 1-3 weeks. The morphological organization of the spinal cord and synaptic inputs onto motoneurons were maintained in organ culture. The rate of differentiation of motoneuron resting potential and increase in membrane excitability was similar in vitro and in vivo, suggesting that these properties were regulated by cellular signals or extracellular differentiation-promoting factors that were preserved in culture. However, maturation of input resistance, action potential threshold and action potential maximum rate of rise was slower than in vivo. Culturing spinal cord explants with their dorsal root ganglia attached did not facilitate motoneuron differentiation. The role of newly formed synaptic pathways in regulating the changes in motoneuron electrical properties was studied in the presence of blockers of synaptic transmission. Motoneuron differentiation was delayed in spinal cords cultured in the presence of TTX, indicating that electrical activity influenced the time course of their development. However, blocking synaptic transmission with antagonists of glutamate, glycine, and GABAA receptors did not affect the rate of motoneuron differentiation, suggesting that maturation of motoneuron phenotype was independent of activation of these transmitter-gated channels. Incubating spinal cords in medium containing high-K+, which increased the frequency of spontaneous potentials, reversed the inhibitory effect of TTX. Similar to TTX action, motoneuron development was retarded when synaptic release was chronically blocked with either tetanus toxin or omega-conotoxin, a Ca2+ channel blocker. These findings suggested that electrical activity in spinal cord explants modulated motoneuron differentiation via Ca(2+)-dependent synaptic release of neurotransmitters or neurotrophic factors.

Animals↗

[Cataract after silicone oil tamponade and its surgical treatment].

Twelve eyes of twelve cases of cataract after complicated retinal detachment surgery and silicone oil tamponade were treated in this report. Successful extracapsular cataract extraction (ECCE) was performed on 10 of the 12 cases, no oil escape occurred in any of the operation and in one eye an intraocular lens was implanted. Intraoperatively, loss of large amount of silicone oil occurred and it was removed completely in ECCE in 2 cases. After the follow-up periods of 3 approximately 30 months (mean, ten months), ten cases with the intact posterior capsule and silicone oil in place had a good view of the fundus, their retinae were attached and visual acuities were recovered above the best levels after retinal detachment surgery. Two cases with silicone oil removal suffered from retinal re-detachment. The main points of the surgery were discussed, and the effective method of prevention from silicone oil escape and maintenance of the transparency of the posterior capsule after ECCE were proposed.

Adolescent↗