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Biomedical subjects

F Zheng

Publications and source records attributed to F Zheng.

At least 91 records · Page 5Linked to original sources
PubMed1997

Evidence that intramolecular interactions are involved in masking the activation domain of transcriptional activator Leu3p.

The Leu3 protein of Saccharomyces cerevisiae regulates the expression of genes involved in branched chain amino acid biosynthesis and in ammonia assimilation. It is modulated by alpha-isopropylmalate, an intermediate in leucine biosynthesis. In the presence of alpha-isopropylmalate, Leu3p is a transcriptional activator. In the absence of the signal molecule, the activation domain is masked, and Leu3p acts as a repressor. The recent discovery that Leu3p retains its regulatory properties when expressed in mammalian cells (Guo, H., and Kohlhaw, G. B. (1996) FEBS Lett. 390, 191-195) suggests that masking and unmasking of the activation domain occur without the participation of auxiliary proteins. Here we present experimental support for this notion and address the mechanism of masking. We show that modulation of Leu3p is exceedingly sensitive to mutations in the activation domain. An activation domain double mutant (D872N/D874N; designated Leu3-dd) was constructed that has the characteristics of a permanently masked activator. Using separately expressed segments containing either the DNA binding domain-middle region or the activation domain of wild type Leu3p (or Leu3-dd) in a modified yeast two-hybrid system, we provide direct evidence for alpha-isopropylmalate-dependent interaction between these segments. Finally, we use the phenotype of Leu3-dd-containing cells (slow growth in the absence of added leucine) to select for suppressor mutations that map to the middle region of Leu3-dd. The properties of nine such suppressors further support the idea that masking is an intramolecular process and suggest a means for mapping the surface involved in masking.

Amino Acid Sequence↗
PubMed1997

The role of paraventricular nucleus of hypothalamus in stress-ulcer formation in rats.

The rat stress model of restraint and cold water immersion was used to investigate the effect of stimulating the paraventricular nucleus (PVN) of hypothalamus on the development of stress-induced gastric ulceration. The results were (1) electric stimulation of the PVN increased the stress ulceration, while electrolytic lesion of the PVN decreased it; (2) intracerebroventricular injection (i.c.v.) of acetylcholine (Ach) enhanced the effect of PVN stimulation on stress ulcers, and the M-receptor was involved; (3) i.c.v. norepinephrine (NE) attenuated the effect of PVN stimulation on stress ulcers in a dose-dependent manner, and the beta-receptor was involved; (4) i.c.v. 5-hydroxytryptamine (5-HT) enhanced the effect of PVN stimulation on stress ulcers; (5) electrolytic lesions of dorsal raphe nucleus (DR) attenuated the effect of PVN stimulation on stress ulcers, while electrolytic lesions of the locus ceruleus (LC) aggravated the effect; (6) thyroidectomy, adrenalectomy, ovariectomy, vagotomy and sympathectomy all attenuated the effect of PVN stimulation on stress ulcers; (7) electric stimulation of the PVN produced no effect on gastric juice volume, acidity, total acid output, pepsin activity or the gastric barrier mucus; but greatly reduced gastric mucosal blood flow. These results indicate that the PVN is an important brain site regulating the development of stress-induced gastric ulcers, that the classical neurotransmitters Ach, NE and 5-HT are involved, and that in the periphery, both the parasympathetic and sympathetic nervous systems and the three endocrine glands (thyroid, adrenal and gonad) take part in the effect.

Acetylcholine↗
PubMed1997

Neurokinin A in rat renal afferent neurons and in nerve fibres within smooth muscle and epithelium of rat and guinea-pig renal pelvis.

Neurokinin A-like immunoreactivity of dorsal root ganglion neurons innervating the kidney were studied with retrograde tracing of FluoroGold dye applied to the cut renal nerves. The proportions and sizes of renal afferent neurons with neurokinin A-like immunoreactivity were quantified in T9-L2 dorsal root ganglia from five rats. Of 240 renal afferent neuronal somata examined, 26 +/- 3% (S.E.M.) showed neurokinin A-like immunoreactivity. Compared with the overall size distribution of renal afferent neurons, those staining for neurokinin A were mostly small-sized neurons with a few medium-sized neurons. All somata with neurokinin A-like immunoreactivity were neurofilament-poor as judged by labelling with an anti-neurofilament antibody, RT97, and it is therefore likely that they had unmyelinated fibres. To examine the sites to which the renal afferent fibres with neurokinin A might project, sections of rat and guinea-pig kidney and upper ureter were examined. Fibres with neurokinin A-like immunoreactivity were found beneath and within the transitional epithelium lining the inner surface of the pelvis, and within the smooth muscle layer beneath the transitional epithelium. Epithelial innervation was found only in regions with underlying smooth muscle and loose connective tissue, and not in sites where the epithelium was closely applied to the renal parenchyma. The network of fibres was most dense towards the pelvo-uretic junction. Fibres with neurokinin A-like immunoreactivity were not seen beneath or within the cuboidal/columnar epithelium covering the papilla within the renal pelvis. Furthermore, only very few fibres with neurokinin A were observed penetrating the transitional epithelium of the upper ureter in both rat and guinea-pig. The distribution of fibres labelled with antibodies to substance P and calcitonin gene-related peptide in the renal pelvis was similar to that for fibres with neurokinin A-like immuno-reactivity, although a few fibres penetrated further into the fornices than fibres with neurokinin-A-like immunoreactivity. Thus, many afferent fibres in the renal pelvis may contain neurokinin A as well as substance P and calcitonin gene-related peptide. These fibres may be the source of the neurokinin A, substance P and calcitonin gene-related peptide which can be released by topical capsaicin treatment. In addition they may be the mechano- and chemo-receptive fibres in the renal pelvis that are known to play important roles in renal haemodynamics. The intra-epithelial position of some of these fibres in the epithelial layer suggests a possible chemosensory or osmosensory role.

Animals↗
PubMed1997

Glomerulosclerosis, arteriosclerosis, and vascular graft stenosis: treatment with oral heparinoids.

At present there is no known treatment for established glomerulosclerosis or atherosclerosis. Since the principal lesion in glomerulosclerosis involves mesangial cells, a vascular smooth muscle cell, we searched for new therapeutic approaches affecting vascular smooth muscle function, especially with respect to modifying the turnover of extracellular matrix. We used mice transgenic for bovine growth hormone (bGH), since these mice develop end-stage renal disease due to progressive glomerulosclerosis. We previously showed that the subcutaneous injection of a non-anticoagulant heparin reduced glomerulosclerosis in bGH mice. Since injectable drugs are not a practical means of controlling glomerulosclerosis in humans, we assessed oral heparin-like compounds and found that oral pentosan polysulfate (PPS) reduced glomerulosclerosis in bGH mice at non-toxic doses. Because the positive therapeutic response in the bGH model could have been principally hormone-mediated, we examined other models of non-immune mediated glomerulosclerosis, including ROP Os/+ non-diabetic and diabetic mice. We found that an oral PPS (Elmiron), which is approved for other indications in humans, reduced sclerosis in all of these forms of chronic, progressive glomerulosclerosis. Based on the similarity of the cellular events in glomerulosclerosis and arteriosclerosis, we assessed the effect(s) of PPS in congenital (Watanabe rabbits) and induced (New Zealand White lipid-fed rabbits) models of arteriosclerosis. The extent and severity of the lesions was significantly reduced in both models by PPS treatment. Finally, we asked whether the proliferative and sclerotic lesion, which is the cause of vascular graft stenosis, might also respond to PPS treatment. To do this we cultured cells from the materials removed from stenotic arteriovenous grafts in hemodialysis patients. We found that PPS inhibits the proliferation and matrix production in a dose-dependent manner.

Animals↗
PubMed1996

Activation of a metabotropic excitatory amino acid receptor potentiates spike-driven calcium increases in neurons of the dorsolateral septum.

(1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), an agonist for metabotropic glutamate receptors (mGluRs), causes depolarization and burst firing in rat dorsolateral septal nucleus (DLSN) neurons and results in long-term potentiation (LTP) at DLSN synapses. In the present study, we investigated whether these actions of 1S,3R-ACPD are attributable to the release of calcium from an inositol triphosphate-sensitive store after activation of mGluRs coupled to phospholipase C. Our data demonstrated that the ACPD-induced depolarization was associated with a small but significant decrease, not an increase, in [Ca2+]i; however, changes of [Ca2+]i, during ACPD-induced bursting were up to seven times larger than those produced by regular firing. Depletion of internal calcium stores by thapsigargin or ryanodine had a small to insignificant effect on the maximum changes of [Ca2+]i, associated with ACPD-induced bursting. Thus, elevation of [Ca2+]i, during firing by 1S,3R-ACPD is likely attributable to enhancement of calcium influx through voltage-gated channels and not to calcium release from internal stores. ACPD-induced burst firing elevated somatic and dendritic calcium levels up to 3 and 6 microM, respectively. Such an increase may be the underlying mechanism for ACPD-induced LTP as well as ACPD-induced acute cell death in rat DLSN.

Action Potentials↗
PubMed1996

Effects of moderate hypothermia on baroreflex and pulmonary chemoreflex heart rate response in decerebrate ferrets.

The effects on the baroreceptor reflexes and pulmonary reflexes of moderate immersion hypothermia (core temperature, 30.2 +/- 0.07 degrees C) have been investigated in the decerebrate ferret. Baroreflex sensitivity was estimated from the relationship between change in cardiac interval and change in systolic blood pressure following i.v. bolus injection of phenylephrine. The responses to phenylephrine in ferrets were best fitted by two linear slopes with an initial shallow slope, a, followed by a steeper slope, b. The slope or sensitivity of b was increased significantly by moderate cooling (n = 6 animals, 36 responses, P < 0.05). Pulmonary J-receptor reflex sensitivity was assessed by i.v. injection of phenylbiguanide (PBG), which evokes a dose-dependent bradycardia. The bradycardic response was unaltered by the hypothermia (n = 6 animals, 88 responses). Electrical stimulation of the right peripheral vagal nerve was employed to assess effects on the efferent components of the reflexes. The bradycardia, in response to stimulation, was significantly increased by moderate cooling at all stimulation frequencies (n = 8 animals, 88 responses, P < 0.001). Thus the results suggested that baroreceptor heart rate reflex sensitivity was enhanced by the moderate hypothermia. At least one component of the enhanced baroreflex response may be the result of changes in the efferent pathway of the reflex response. However, the absence of effect of PBG may indicate a differential afferent and efferent organization of pulmonary J-receptors compared with baroreceptors.

Animals↗
PubMed1996

[Immune-mediated tubulointerstitial injury in hemorrhagic fever with renal syndrome].

OBJECTIVE: To study the presence of Hantan virus RNA and specific immune-complex deposits in the renal tubular epithelium of HFRS patients. METHODS: Renal biopsy specimens from 15 cases of HFRS, 3 cases of ischemic ATN and 5 normal controls (donor of transplantation) were investigated. Human HLA-DR, ICAM-ILFA-1 and CD4+, CD8+, CD8+ cells in the renal tissue section were quatitatively analyzed by immuno-peroxidase staining 4 layers PAP. Histo-morphometric analysis of pathological section was made for each specimen. RESULTS: HFRS presented abundant amount of HLA-DR and ICAM-1 in tubular cells both on cortical and medullary region, while ischemic ATN tended to have such positive results only in cells of cortical region. The intensity and extent of HLA-DR and ICAM-1 LFA-1 staining were closely correlated with the degree of tubulointerstitial damage of HFRS. Interstitial CD4, CD8 and monocytes increased in amount profoundly in HFRS with a preponderance in the medullary region, and particularly of CD8 positive cells. Less marked increment occurred in ATN cases, and no discrepancies were found between cortical and medullary region or cell types. The severity of tubulointerstitial lesion was closely associated with the extent of cellular infiltration and the expression of HLA-DR, ICAM-1 in renal tubular cells. CONCLUSION: The characteristic changes of tubulointerstitial lesions in HFRS suggest that virus infection may be one of the mediators responsible for the immune kidney injuries in HFRS.

Adult↗
PubMed1996

[Impact of platelet derived growth factor in the glomeruli of active lupus nephritis].

Platelet derived growth factor (PDGF) possesses diverse biological activities and plays an important role in the pathogenesis of glomeruli nephritis. In order to elucidate the relationship between PDGF and the disease activation in lupus nephritis, PDGF was observed in renal biopsy specimen from 9 cases of lupus nephritis (7 cases with active and 2 cases with inactive lesions) by immunohistochemistry using 4-layer PAP method. The mRNA expression of PDGF-A and -B chain, and the receptor of PDGF-B were analyzed by RT-PCR. It was found that the levels of mRNA expression of PDGF and its receptor were much higher in patients with active lesions in glomeruli as compared with those without active lesions, there changes were paralleled with the degree of hematuria in these patients. Noted that there was no correlation between the expression of PDGF and the cellular proliferation in glomeruli of lupus nephritis. These results indicate that PDGF is a critical mediator for inducing active lesion in glomeruli of lupus nephritis. The effect of PDGF antagonist in the regulation of glomerular damage in lupus nephritis need to further elucidate.

Adolescent↗
PubMed1996

[Experimental study on effect of Cordyceps sinensis in ameliorating aminoglycoside induced nephrotoxicity].

In order to evaluate the effect of Cordyceps sinensis (CS) on aminoglycoside (AG) induced nephrotoxicity, gentamycin was imposed on the young and old rats with CS administration. The renal tubular injury was ameliorated as evidenced by less prominent increment of BUN, SCr, sodium excretion, urinary NAGase and less severity of histopathological changes as compared with control. In addition, the use of CS could promote an earlier recovery of renal oxygen consumption insulin clearance, and sodium absorption in isolated perfused kidney from CS treated intoxicated rat than that from control. Possible mechanisms of CS on drug-induced nephrotoxicity include: (1) Accelerating the regeneration of tubular cells; (2) Protecting the sodium pump activity of tubular cells; (3) Attenuating the tubular cell lysosome hyperfunction stimulated by phagocytosis of AG as well as decreasing the tubular cell lipoperoxidation in response to toxic injury; (4) Reducing the tissue Ca++ content.

Acute Kidney Injury↗
PubMed1995

1S,3R-ACPD-preferring inward current in rat dorsolateral septal neurons is mediated by a novel excitatory amino acid receptor.

Metabotropic glutamate receptors (mGluRs) form a receptor family that consists of diverse receptor subtypes; now, numbering 8--exclusive of splice variants. (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) has been suggested to be a selective agonist for the mGluRs. We have recently reported that, in rat dorsolateral septal nucleus (DLSN) neurones, a 1S,3R-ACPD-preferring inward current (ACPDi) persists in pertussis toxin-treated rats. We now report that this ACPDi-current: (1) persists in DLSN neurones dialyzed with a stable analog of GTP, namely, GTP gamma S; (2) exhibits a negative slope region with inward rectification in its I-V relationship; (3) persists in neurones superfused with tetrodotoxin or low calcium solutions; (4) is dependent upon both sodium and calcium ions; and (5) is independent of a reduction in temperature. Furthermore, pharmacological data suggest that this current may be activated by a unique type of excitatory amino acid (EAA) receptor, i.e. a receptor which prefers "metabotropic" EAA agonists and is insensitive to AP5 or CNQX. Activation by ACPD of inward currents associated with a conductance increase have also been reported at cultured mouse cerebellar Purkinje neurones; in slices of rat hippocampal CA1 neurones and slice cultures of hippocampal CA3 neurones. We suggest that this ACPDi current may play an important role within the CNS in the induction of long-term potentiation and other neurological processes; processes attributed previously to currents associated with NMDA receptor activation.

2-Amino-5-phosphonovalerate↗
PubMed1995

(1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid-induced burst firing is mediated by a native pertussis toxin-sensitive metabotropic receptor at rat dorsolateral septal nucleus neurons.

We have reported previously that a selective metabotropic glutamate receptor agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), caused two primary postsynaptic membrane changes, namely, a slow membrane depolarization, and burst firing in rat dorsolateral septal nucleus neurons. In addition, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid also potentiates a slow after depolarization in rat dorsolateral septal nucleus neurons. We now report that, among all the postsynaptic membrane changes induced by (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid, only the burst firing was selectively blocked by pertussis toxin pretreatment. Thus, aminocyclopentane-1,3-dicarboxylic acid induced burst firing was mediated by a metabotropic receptor coupled to a pertussis toxin-sensitive GTP-binding protein, while the other induced cellular responses may be mediated by metabotropic glutamate receptors insensitive to pertussis toxin. We further characterized this receptor pharmacologically. This metabotropic receptor is activated by several metabotropic glutamate receptor agonists, but is insensitive to L-glutamate or L-aspartate. On the basis of its agonist activity profile, particularly the ineffectiveness of glutamate as an agonist, we have tentatively assigned the name aminocyclopentane-1,3-dicarboxylic acid metabotropic receptor, to this native, pertussis toxin-sensitive metabotropic receptor in the dorsolateral septal nucleus. Furthermore, this receptor is coupled to protein kinase C, probably via a phospholipase C independent pathway.

Animals↗
PubMed1995

[Different expressions of alpha 2 (IV) and alpha 3 (IV) collagen mRNAs in renal glomeruli of IgA nephropathy].

The levels of extracellular matrix protein mRNAs were increased prior to the appearance of histological glomerulosclerosis in isolated glomeruli from several animal models. Striker studied renal biopsies (Seminar in Nephrology, 13:508,1993). We made a comparable study in human renal biopsies to assess the levels of glomerular alpha 2 (IV) and alpha 3 (IV) collagen gene expressions in renal biopsies. Renal biopsy specimens were obtained from 11 patients (7 IgA nephropathy and 4 mesangial proliferative glomerulonephritis). Glomerular lesion was divided into 5 grades. The amount of collagen type IV in glomerular matrix was evaluated by immunoperoxidase staining using a monoclonal antibody to type IV collagen. For gene expression analysis, glomeruli were dissected out from about 10% of each biopsy specimen. The expressions of alpha 2 (IV) and alpha 3 (IV) collagen mRNAs were analyzed by using in situ reverse transcription coupled with polymerase chain reaction. Each PCR reaction contained an amount of cDNA template equivalent to that obtained from 1/10 of a glomerulus, alpha 2 (IV) and alpha 3 (IV) collagen mRNAs were detected in all specimens. All 3 cases of IgAN with grade III glomerular lesion had a more marked increment of glomerular alpha 2 (IV) collagen cDNA than that of the other 3 IgAN patients with grade II lesion (the intensity of PCR product, grade III 0.54 +/- 0.12, VS grade II 0.25 +/- 0.08, P < 0.05). No discrepancies were detected in the level of glomerular alpha 3 (IV) collagen cDNA and the intensity of collagen IV staining between these two groups of IgAN. Interestingly, in MsPGN patients there was no difference in the expression of glomerular alpha 2 (IV) collagen mRNA between grade II and grade III glomerular lesions. The results suggested that the detection of glomerular alpha 2 (IV) collagen mRNA level in IgAN patients may help to define the pathological classification.

Adolescent↗
PubMed1995

Pharmacologically distinct, pertussis toxin-resistant inward currents evoked by metabotropic glutamate receptor (mGluR) agonists in dorsolateral septal nucleus (DLSN) neurons.

We have reported previously that a selective metabotropic glutamate receptor (mGluR) agonist, (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), caused a slow membrane depolarization in rat dorsolateral septal nucleus (DLSN) neurons. Using single electrode voltage-clamp recording methods, we now investigate the pharmacological properties of the receptor that mediates ACPD-induced membrane currents in DLSN neurons recorded from pertussis toxin (PTX)-treated rats. Two pharmacologically distinct inward currents, that is, the ACPD current and Qm current, have been identified based on their agonist preference and sensitivity to various antagonists. The ACPD current is blocked by L-2-amino-4-phosphonobutyric acid (L-AP4), but is insensitive to L-aspartic acid-beta-hydroxamate (L-AA beta H), (+)-alpha-methyl-4-carboxyphenylglycine (+)-MCPG), or L-2-amino-3-phosphonopropionic acid (L-AP3). The Qm current is blocked by L-AA beta H and (+)-MCPG, but is insensitive to L-AP3 or L-AP4. These two inward currents distribute differentially within subpopulations of DLSN neurons. The ACPD current is the only current observed in most DLSN "burster" neurons, while the Qm current is observed more frequently in DLSN "nonburster" neurons. The pharmacological profiles of these currents suggest that the Qm current is likely mediated by mGluR1 or mGluR5, while the ACPD current is mediated by receptors that are pharmacologically distinct from any of the currently cloned mGluRs.

Animals↗
PubMed1994

(1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) induces burst firing via an inositol-1,4,5-triphosphate-independent pathway at rat dorsolateral septal nucleus.

We have previously reported that a L-2-amino-3-phosphonopropionate (L-AP3)-sensitive metabotropic glutamate receptor was required for the induction of long-term potentiation (LTP) in rat dorsolateral septal nucleus neurons. (1S,3R)-1-Aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD), a selective agonist for metabotropic glutamate receptors, also causes burst firing of dorsolateral septal nucleus (DLSN) neurons. In this study, we investigated whether this response was mediated by a phospholipase C-(PLC) coupled metabotropic glutamate receptor. The threshold concentration of 1S,3R-ACPD for the induction of burst firing was about 5 microM, while 10 microM 1S,3R-ACPD produced a maximal effect. L-AP3 (50 microM) reduced the burst firing induced by 1S,3R-ACPD (5 microM). Although 1S,3R-ACPD stimulated the formation of inositol-1,4,5-triphosphate [Ins(1,4,5)P3] suggesting the presence of PLC-coupled metabotropic glutamate receptors, it was only effective in a higher (30-100 microM) concentration range. In addition, the 1S,3R-ACPD-stimulated formation of Ins(1,4,5)P3 level was not affected by L-AP3. These observations suggest that the 1S,3R-ACPD induced burst firing is not mediated by PLC-coupled metabotropic glutamate receptors.

Action Potentials↗