Biomedical subjects

H Zhong

Publications and source records attributed to H Zhong.

At least 109 records · Page 6Linked to original sources

PubMed1998

Serum vascular endothelial growth factor is a candidate biomarker of metastatic tumor response to ex vivo gene therapy of renal cell cancer.

We report the close correlation between changes in serum immunoreactive vascular endothelial growth factor 165 (iVEGF165) levels and metastatic tumor burden measured by computed tomography scan before treatment, during the antitumor response, and during early progression in a patient treated with ex vivo gene therapy for renal cell carcinoma. With the researcher blinded to outcome, iVEGF levels were measured in archived serum samples from a patient with metastatic renal cell carcinoma who demonstrated a 7-month partial remission to treatment with autologous, irradiated human GM-CSF gene transduced tumor vaccine. Although a spontaneous regression could not be formally excluded in this patient, the appearance of 20 new pulmonary metastases on computed tomography scan after nephrectomy and before vaccination indicates that if spontaneous regression occurred, it took place at the start of vaccine treatment.

Biomarkers, Tumor↗

PubMed1998

Phosphorylation of NF-kappa B p65 by PKA stimulates transcriptional activity by promoting a novel bivalent interaction with the coactivator CBP/p300.

The transcriptional activity of NF-kappa B is stimulated upon phosphorylation of its p65 subunit on serine 276 by protein kinase A (PKA). The transcriptional coactivator CPB/p300 associates with NF-kappa B p65 through two sites, an N-terminal domain that interacts with the C-terminal region of unphosphorylated p65, and a second domain that only interacts with p65 phosphorylated on serine 276. Accessibility to both sites is blocked in unphosphorylated p65 through an intramolecular masking of the N terminus by the C-terminal region of p65. Phosphorylation by PKA both weakens the interaction between the N- and C-terminal regions of p65 and creates an additional site for interaction with CBP/p300. Therefore, PKA regulates the transcriptional activity of NF-kappa B by modulating its interaction with CBP/p300.

Acetyltransferases↗

PubMed1998

Cyclical passive stretch influences the mechanical properties of the inactive cat soleus.

The effects of cyclical, passive manipulation (PM, 30 min day(-1), 5 days week(-1) for 6 months) mimicking the length excursions observed during stepping on the mechanical and associated biochemical properties of the inactive cat soleus muscle were determined in five cats. Inactivity was produced via spinal cord isolation (SI), i.e. complete spinal cord transections at low thoracic and high sacral levels and bilateral dorsal rhizotomy between the transection sites. Passive manipulation was administered to one leg of each SI cat. Compared with normal controls, SI resulted in approximately 70% decrease in weight, an 80% decrease in maximum tetanic tension (Po) and an approximately 100% increase in maximum rate of shortening (Vmax) and myosin adenosine triphosphatase (mATPase) activity of the soleus. The passive manipulation regime partially ameliorated these effects. When compared with the control SI soleus, the SI-PM soleus weight and maximum tetanic tension were 12 and 21% higher, respectively, and the Vmax and mATPase activity 21 and 12% (p > 0.05) lower, respectively. Thus, inactivity resulted in a smaller and faster muscle, whereas passive manipulation for only 30 min a day tended to maintain these properties closer to normal control values. The results suggest a potential therapeutic effect of short bouts of cyclical, passive manipulation on otherwise inactive skeletal muscles.

Animals↗

PubMed1998

Skeletal muscle afferent regulation of bioassayable growth hormone in the rat pituitary.

There are forms of growth hormone (GH) in the plasma and pituitary of the rat and in the plasma of humans that are undetected by presently available immunoassays (iGH) but can be measured by bioassay (bGH). Although the regulation of iGH release is well documented, the mechanism(s) of bGH release is unclear. On the basis of changes in bGH and iGH secretion in rats that had been exposed to microgravity conditions, we hypothesized that neural afferents play a role in regulating the release of these hormones. To examine whether bGH secretion can be modulated by afferent input from skeletal muscle, the proximal or distal ends of severed hindlimb fast muscle nerves were stimulated ( approximately 2 times threshold) in anesthetized rats. Plasma bGH increased approximately 250%, and pituitary bGH decreased approximately 60% after proximal nerve trunk stimulation. The bGH response was independent of muscle mass or whether the muscles were flexors or extensors. Distal nerve stimulation had little or no effect on plasma or pituitary bGH. Plasma iGH concentrations were unchanged after proximal nerve stimulation. Although there may be multiple regulatory mechanisms of bGH, the present results demonstrate that the activation of low-threshold afferents from fast skeletal muscles can play a regulatory role in the release of bGH, but not iGH, from the pituitary in anesthetized rats.

Animals↗

PubMed1998

[One stage repair of old Achilles tendon rupture accompanied by skin defect].

Old achilles tendon rupture accompanied by skin defect was a common & annoying problem in clinic. From June, 1985 to June, 1996, 18 cases with this kind of injury were treated by one stage repair of the tendon and skin defect. In this series, there were 15 males and 3 females, the length of tendon defects were ranged from 4 cm-6.1 cm, and the area of skin defect were ranged from 5.9 cm x 3 cm to 8.2 cm x 6 cm. The procedures were: (1) to debridement of the wound thoroughly; (2) to repair the achilles tendon; (3) to repair the skin defect with kinds of pedicle flap; (4) immobilization of ankle and knee for 6 weeks. No infection was occured after the operation. The flaps survived in all cases. After follow-up for one year in 15 cases, 12 patients went back to their work. It was concluded that (1) achilles tendon rupture should be treated carefully and properly during the emergency operation; (2) different methods should be selected according to the length of tendon defect; (3) because of its high survival and retained sensation after operation, the flap pedicled with posterior lateral malleolar artery is the best choice for repairing the skin defect.

Achilles Tendon↗

PubMed1998

[Experimental study on hypoglycemic effect of YTY granules].

OBJECT: To perform experimental pharmacodynamic evaluation of a new TCM antidiabetic drug Yitangyin (YTY). METHODS: Experimkental streptozotocin induced rat diabetes models were subjected YTY treatment and the hypoglycemic effect was evaluated. The proved TCM Yusanxiao (YSX) was used as contrast. RESULTS: The low, middle and high dose YTY groups exhibited significant hypoglycemic effects on the model rats. The efficiency was comparable with YSX. CONCLUSION: The TCM YTY granule is an effective hypoglycmeic agent.

Animals↗

PubMed1998

[A study on the correlation between the activity of 72,000 type IV collagenase and the metastatic potential of cancer cells].

OBJECTIVE: To evaluate the enzyme activity of 72,000 type IV collagenase and its relationship with the metastatic potential of cancer cells. METHODS: The levels of secreted 72,000 type IV collagenase in the conditioned media of five human cancer cell lines with different metastatic potential and a normal lung fibroblast strain treated with cancer cell culture media were examined by gelatin zymography and densitometric analysis. RESULTS: The levels of 72,000 type IV collagenase secreted by cancer cells with high metastatic potential (PG, WM451 and WM983a) were higher than those secreted by cancer cells with low metastatic potential (PAa and WM35). In the conditioned media of fibroblasts which were treated with the culture media of PG and WM451, enhanced levels of activation were observed. CONCLUSION: The secretion of 72,000 type IV collagenase is closely correlated to the metastatic potential of cancer cells. The cancer cells with high metastatic potential may possibly through certain soluble mediators stimulate normal fibroblasts to activate 72,000 type IV procollagenase.

Adenocarcinoma↗

PubMed1998

[Study of protein separation by reversed-phase high performance liquid chromatography].

Reversed-phase high performance liquid chromatography (RP-HPLC) of protein on large pore silica (20-30 nm) n-alkyl-bonded(C4 and C5) phase columns was studied using a trifluoroacetic acid (TFA)/2-propanol mobile phase system. The influence of concentration of organic modifier (2-propanol) and ion-pairing acid (TFA) on the protein retention time was examined. The results showed that the plot of the retention time vs-2-propanol concentration in mobile phase was U-shaped. The mechanism of protein in reversed-phase high performance liquid chromatography is also discussed.

Alpha-Globulins↗

PubMed1998

[Reliability design and analysis of man-machine performance of display and illumination system in cockpit].

This paper analysed and discussed the basic reliability of display and illumination system in which Chinese characters displayed on LCD were read and the colours were identified, and instruments in different visual views and the instruments of differential display models were read. The colour rendering of the light source was also evaluated. Basic reliability may be chosen as 0.9995.

Aircraft↗

PubMed1997

Synapse formation and hypoxic signalling in co-cultures of rat petrosal neurones and carotid body type 1 cells.

1. To investigate synaptic mechanisms mediating chemosensory signalling in the carotid body, we developed co-cultures of chemoreceptor type 1 cell clusters and dissociated petrosal neurones (PNs) from 7- to 14-day-old rat pups and tested for functional connectivity in CO2-HCO3(-)-or Hepes-buffered medium at approximately 35 degrees C. 2. When cultured without type 1 cells, PNs were almost always quiescent (n = 104) and unresponsive to hypoxia (Po2 = 5-25 mmHg) during perforated patch, whole-cell recordings of membrane potential or voltage-activated currents; in contrast, many PNs (77 out of 170) that were juxtaposed to type 1 cell clusters in co-culture displayed spontaneous activity, comprising spikes and subthreshold potentials (SSPs) that resembled synaptic potentials. 3. Additional tests suggested that de novo chemical synapses developed between PNs and type 1 cell clusters in vitro. For example: (i) the spontaneous activity was reversibly suppressed by substituting low calcium-high magnesium in the bath; (ii) SSPs had variable amplitudes and persisted following action potential blockade with TTX (1 microM); (iii) the interval distribution between successive spontaneous events appeared random; and (iv) the frequency of spontaneous potentials was diminished (reversibly) by the nicotinic antagonist hexamethonium (100 microM), suggesting contributions from the spontaneous release of ACh. 4. Many complexes of 'juxtaposed' PNs and type 1 clusters were physiologically functional, since exposure to hypoxia caused a reversible depolarization and/or increased spike discharge in approximately 30% of such neurones (n = 140). The hypoxia-induced spike discharge persisted in the presence of the dopamine D2 receptor blocker spiperone (10-50 microM; n = 5); however, this discharge was reversibly inhibited by 100-200 microM hexamethonium, suggesting that it was mediated, at least in part, by ACh acting through nicotinic receptors. 5. The hypoxia-induced spike discharge and frequency of spontaneous potentials in co-cultured PNs were reversibly suppressed when the buffer was switched from CO2-HCO3- to Hepes (10 mM) at pH 7.4; further, 'functional' PNs that displayed spontaneous activity and/or hypoxia-induced responses in co-culture were encountered more frequently in CO2-HCO3- (> or = 40%) than in Hepes (< or = 26%) buffer. 6. We conclude that functional chemical synapses can develop de novo in cultures of carotid body type 1 cells and PNs and that ACh is probably an important excitatory neurotransmitter secreted from type 1 cells during hypoxic chemotransduction in the rat carotid body.

Acetylcholine↗

PubMed1997

Baculovirus p35 and Z-VAD-fmk inhibit thapsigargin-induced apoptosis of breast cancer cells.

Programmed cell death, or apoptosis, is inhibited by the antiapoptotic oncogene, Bcl-2, and is mediated by a cascade of aspartate-specific cysteine proteases, or caspases, related to interleukin 1-beta converting enzyme. Depending on cell type, apoptosis can be induced by treatment with thapsigargin (TG); a selective inhibitor of the endoplasmic reticulum-associated calcium-ATPase. The role of caspases in mediating TG-induced apoptosis was investigated in the Bcl-2-negative human breast cancer cell line, MDA-MB-468. Apoptosis developed in MDA-MB-468 cells over a period of 24-72 h following treatment with 100 nM TG, and was prevented by Bcl-2 overexpression. TG-induced apoptosis was associated with activation of caspase-3 and was inhibited by stable expression of the baculovirus p35 protein, an inhibitor of caspase activity. Also, TG-induced apoptosis was inhibited by treating cells with Z-VAD-fmk, a cell-permeable fluoromethylketone inhibitor of caspases. These findings indicate that TG-induced apoptosis of MDA-MB-468 breast cancer cells is subject to inhibition by Bcl-2 and is mediated by caspase activity. This model system should be useful for further investigation directed toward understanding the role of calcium in signaling apoptosis, and its relationship to Bcl-2 and the caspase proteolytic cascade.

Amino Acid Chloromethyl Ketones↗

PubMed1997

Nicotinic acetylcholine sensitivity of rat petrosal sensory neurons in dissociated cell culture.

Using whole-cell, patch-clamp techniques we investigated acetylcholine (ACh) sensitivity of dissociated sensory neurons from rat petrosal ganglia after 4 h-14 days in vitro. In approx. 68% of petrosal neurons (PN; n = 109) ACh, applied by fast perfusion or pressure ejection from a 'puffer' pipette, caused a rapid depolarization associated with a conductance increase. Under voltage clamp near the resting potential (approx. - 60 mV), ACh induced a hexamethonium-sensitive, inward current (IACh), mimicked by nicotine application, suggesting the presence of neuronal nicotinic acetylcholine receptors (nAChR). The reversal potential of IACh occurred near 0 mV (n = 4), a region where the I-V curve displayed a prominent rectification. The dose-response relation for IACh versus ACh concentration was fitted by the Hill equation with EC50 = approx. 33.9 microM and Hill coefficient = approx. 1.6. The activation phase of IACh was well fitted by a single exponential with mean (+/- S.E.M.) time constant of 102 +/- 82 ms (n = 6); the desensitization phase of IACh was best fitted by the sum of two exponentials, with time constant of 870 +/- 210 ms (n = 6) and 8576 +/- 1435 ms (at -70 mV). Fluctuation analysis yielded an apparent single-channel conductance of 21.6 +/- 10 pS (mean +/- S.E.M.; n = 4). These data indicate that a major subpopulation of sensory neurons in visceral petrosal ganglia of the rat express nAChR. Thus, if similar receptors are present on corresponding nerve terminals, they could mediate fast afferent excitation in response to ACh released at peripheral targets, e.g., the chemosensory carotid body.

Acetylcholine↗

PubMed1997

Alpha2p controls donor preference during mating type interconversion in yeast by inactivating a recombinational enhancer of chromosome III.

Homothallic strains of Saccharomyces cerevisiae can change mating type as often as every generation by replacing the allele at the MAT locus with a copy of mating type information present at one of two storage loci, HML and HMR, located on either end of chromosome III. Selection of the appropriate donor locus is dictated by a mating type-specific repressor protein, alpha2p: Cells containing alpha2p select HMR, whereas those lacking alpha2p select HML. As a repressor protein, alpha2p binds to DNA cooperatively with the transcriptional activator Mcm1p. Here we show that two alpha2p/Mcm1p-binding sites, DPS1 and DPS2, control donor selection. DPS1 and DPS2 are located approximately 30 kb from the left arm of chromosome III, well removed from HML, HMR, and MAT. Precise deletion of only DPS1 and DPS2 results in random selection of donor loci and in a cells without affecting selection in alpha cells. Reciprocally, deletion of only the alpha2p binding segments in each of these two sites results in selection of the wrong donor loci in alpha cells without affecting preference in a cells. These results suggest that Mcm1p, bound to these two sites in the absence of alpha2p, activates HML as donor. Binding of alpha2p blocks the ability of Mcm1p bound to DPS1 and DPS2 to activate HML, resulting in default selection of HMR as donor. DPS1 and DPS2 also regulate expression of several noncoding RNAs, although deletion of at least one of these RNA loci does not affect donor preference. This suggests that transcriptional activation, rather than transcription of a specific product, is the initiating event in activating the left arm of chromosome III for donor selection.

Binding Sites↗

PubMed1997

Chronic activation of protein kinase C by phorbol ester reduces calcium channel expression in chick sympathetic neurons.

Chronic activation of protein kinase C (PKC) has been implicated in regulation of Ca2+ entry responsible for normal development of transmitter properties in cultured sympathetic neurons. The idea that PKC alters the expression of Ca2+ channels was tested using phorbol 12,13-dibutyrate (PDB) which activates PKC and also supports survival of chick sympathetic neurons in the absence of nerve growth factor (NGF). Whole cell voltage-clamp showed that neurons supported by PDB for 2 days had significantly lower Ca2+ current density (0.243 +/- 0.025 pA/microm2) than those supported by NGF (0.356 +/- 0.033 pA/microm2). [125I]omega-Conotoxin GVIA binding showed that PDB-supported neurons had significantly lower maximum binding (617 +/- 223 fmol/mg protein) compared with those supported by NGF (1099 +/- 192 fmol/mg protein). These results support the conclusion that chronic activation of PKC limits the expression of N-type Ca2+ channels. A reduction in Ca2+ channel number is consistent with, and could account for the mature type Ca2+ handling and transmitter release properties seen in sympathetic neuro-effector preparations, sympathetic neurons co-cultured with their targets, and neurons supported by PDB.

Animals↗

PubMed1997

The transcriptional activity of NF-kappaB is regulated by the IkappaB-associated PKAc subunit through a cyclic AMP-independent mechanism.

Stimulation of cells with inducers of NF-kappaB such as LPS and IL-1 leads to the degradation of IkappaB-alpha and IkappaB-beta proteins and translocation of NF-kappaB to the nucleus. We now demonstrate that, besides the physical partitioning of inactive NF-kappaB to the cytosol, the transcriptional activity of NF-kappaB is regulated through phosphorylation of NF-kappaB p65 by protein kinase A (PKA). The catalytic subunit of PKA (PKAc) is maintained in an inactive state through association with IkappaB-alpha or IkappaB-beta in an NF-kappaB-IkappaB-PKAc complex. Signals that cause the degradation of IkappaB result in activation of PKAc in a cAMP-independent manner and the subsequent phosphorylation of p65. Therefore, this pathway represents a novel mechanism for the cAMP-independent activation of PKA and the regulation of NF-kappaB activity.

Adenosine Triphosphate↗

PubMed1997

The yeast homeodomain protein MATalpha2 shows extended DNA binding specificity in complex with Mcm1.

The MATalpha2 (alpha2) repressor interacts with the Mcm1 protein to turn off a-cell type-specific genes in the yeast Saccharomyces cerevisiae. We compared five natural alpha2-Mcm1 sites with an alpha2-Mcm1 symmetric consensus site (AMSC) for their relative strength of repression and found that the AMSC functions slightly better than any of the natural sites. To further investigate the DNA binding specificity of alpha2 in complex with Mcm1, symmetric substitutions at each position in the alpha2 half-sites of AMSC were constructed and assayed for their effect on repression in vivo and DNA binding affinity in vitro. As expected, substitutions at positions in which there are base-specific contacts decrease the level of repression. Interestingly, substitutions at other positions, in which there are no apparent base-specific contacts made by the protein in the alpha2-DNA co-crystal structure, also significantly decrease repression. As an alternative method to examining the DNA binding specificity of alpha2, we performed in vitro alpha2 binding site selection experiments in the presence and absence of Mcm1. In the presence of Mcm1, the consensus sequences obtained were extended and more closely related to the natural alpha2 sites than the consensus sequence obtained in the absence of Mcm1. These results demonstrate that in the presence of Mcm1 the sequence specificity of alpha2 is extended to these positions.

ATP-Binding Cassette Transporters↗

PubMed1997

DNA-binding specificity of Mcm1: operator mutations that alter DNA-bending and transcriptional activities by a MADS box protein.

The yeast Mcm1 protein is a member of the MADS box family of transcriptional regulatory factors, a class of DNA-binding proteins found in such diverse organisms as yeast, plants, flies, and humans. To explore the protein-DNA interactions of Mcm1 in vivo and in vitro, we have introduced an extensive series of base pair substitutions into an Mcm1 operator site and examined their effects on Mcm1-mediated transcriptional regulation and DNA-binding affinity. Our results show that Mcm1 uses a mechanism to contact the DNA that has some significant differences from the one used by the human serum response factor (SRF), a closely related MADS box protein in which the three-dimensional structure has been determined. One major difference is that 5-bromouracil-mediated photo-cross-linking experiments indicate that Mcm1 is in close proximity to functional groups in the major groove at the center of the recognition site whereas the SRF protein did not exhibit this characteristic. A more significant difference is that mutations at a position outside of the conserved CC(A/T)6GG site significantly reduce Mcm1-dependent DNA bending, while these substitutions have no effect on DNA bending by SRF. This result shows that the DNA bending by Mcm1 is sequence dependent and that the base-specific requirements for bending differ between Mcm1 and SRF. Interestingly, although these substitutions have a large effect on DNA bending and transcriptional activation by Mcm1, they have a relatively small effect on the DNA-binding affinity of the protein. This result suggests that the degree of DNA bending is important for transcriptional activation by Mcm1.

Amino Acid Sequence↗

PubMed1997

Architectural and histochemical properties of cat hip 'cuff' muscles.

The architectural properties and fiber-type composition of the cat hip 'cuff' muscles, i.e. the deep layer of muscles encircling the hip joint, were determined. The muscles studied included the gluteus minimus, obturator internus and externus, gemellus superior and inferior, pyriformis, quadratus femoris and capsularis. In addition, the fiber-type composition was determined for the iliacus muscle. Compared to other lower limb muscles, the cuff muscles were small (approximately 0.2 to 2.7 g) and short (approximately 15 to 40 mm long) and had short mean fiber lengths (approximately 7 to 12 mm long), small angles of fiber pinnation (< or = 12 degrees), and small physiological cross-sectional areas (approximately 0.2 to 2.8 cm2). The percentage of the cross-sectional area comprised of slow fibers ranged from 24 to 95% with this value being over 50% in 5/9 muscles studied. The small angle of pinnation and short fiber lengths optimize force production and the relatively high percentage of slow fibers suggest a high level of activation. Both of these properties are consistent with a hip stabilization role for these muscles. In addition, the high percentage of slow fibers suggests a high spindle density and a possible role of these muscles in providing proprioceptive feedback to the central nervous system for the control of posture and locomotion.

Animals↗