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Biomedical subjects

Y Zhong

Publications and source records attributed to Y Zhong.

At least 37 records · Page 2Linked to original sources
PubMed2001

P2X receptors in peripheral neurons.

P2X receptors are a family of ligand-gated ion channels, activated by extracellular ATP. The seven subunits cloned (P2X1-7) can assemble to form homomeric and heteromeric receptors. Peripheral neurons of neural crest origin (e.g. those in dorsal root, trigeminal, sympathetic and enteric ganglia) and placodal origin (e.g. those in nodose and petrosal ganglia) express mRNAs for multiple P2X subunits. In this review, we summarize the molecular biological, electrophysiological and immunohistochemical evidence for P2X receptor subunits in sensory, sympathetic, parasympathetic, pelvic and myenteric neurons and adrenomedullary chromaffin cells. We consider the pharmacological properties of these native P2X receptors and their physiological roles. The responses of peripheral neurons to ATP show considerable heterogeneity between cells in the same ganglia, between ganglia and between species. Nevertheless, these responses can all be accounted for by the presence of P2X2 and P2X3 subunits, giving rise to varying proportions of homomeric and heteromeric receptors. While dorsal root ganglion neurons express predominantly P2X3 and rat sympathetic neurons express mainly P2X2 receptors, nodose and guinea-pig sympathetic neurons express mixed populations of P2X2 and heteromeric P2X2/3 receptors. P2X receptors are important for synaptic transmission in enteric ganglia, although their roles in sympathetic and parasympathetic ganglia are less clear. Their presence on sensory neurons is essential for some processes including detection of filling of the urinary bladder. The regulation of P2X receptor expression in development and in pathological conditions, along with the interactions between purinergic and other signalling systems, may reveal further physiological roles for P2X receptors in autonomic and sensory ganglia.

Animals↗
PubMed2001

Genetic manipulation of the odor-evoked distributed neural activity in the Drosophila mushroom body.

Odor-induced neural activity was recorded by Ca2+ imaging in the cell body region of the Drosophila mushroom body (MB), which is the second relay of the olfactory central nervous system. The signals recorded are mainly from the cell layers on the brain surface because of the limited penetration of Ca2+-sensitive dyes. The densely packed cell bodies and their accessibility allow visualization of odor-induced population neural activity. It is revealed that odors evoke diffused neural activities in the MB. Although the signals cannot be attributed to individual neurons, patterns of the population neural activity can be analyzed. The activity pattern, but not the amplitude, of an odor-induced population response is specific for the chemical identity of an odor and its concentration. The distribution pattern of neural activity can be altered specifically by genetic manipulation of an odor binding protein and this alteration is closely associated with a behavioral defect of odor preference. These results suggest that the spatial pattern of the distributed neural activity may contribute to coding of odor information at the second relay of the olfactory system.

Acetates↗
PubMed2001

Multiple P2X receptors on guinea-pig pelvic ganglion neurons exhibit novel pharmacological properties.

1. Application of ATP and alpha,beta-methylene ATP (alpha beta meATP) to voltage-clamped guinea-pig pelvic neurons produced three types of inward currents. A fast-desensitizing response was present in 5% (25/660) of neurons, 70% gave slowly-desensitizing currents, and the remainder had biphasic responses. 2. Slowly-desensitizing responses were characterized pharmacologically. The response to alpha beta meATP 100 microM was 46+/-27% (range 0--100%) of that evoked by ATP 100 microM in the same cell. Cross-desensitization indicated the presence of alpha beta meATP-sensitive and -insensitive receptors. 3. The concentration-response curve for alpha beta meATP had an EC(50) of 55 microM, and a Hill coefficient of 0.99, while at the alpha beta meATP-insensitive receptor, ATP had an EC(50) of 73 microM, with a Hill coefficient of 1.78. 4. The response to alpha beta meATP was blocked by pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), suramin and Cibacron blue. However, the alpha beta meATP-insensitive receptor was inhibited by PPADS, but not by the other two antagonists. 5. 2'- (or 3'-) O-trinitrophenyl-ATP was 10 times more potent in inhibiting responses to alpha beta meATP than to ATP (at the alpha beta meATP-insensitive receptor). 6. Lowering extracellular pH potentiated responses to alpha beta meATP and ATP, while raising pH attenuated them. 7. Co-application of Zn(2+) (3--300 microM) inhibited the responses to alpha beta meATP and ATP, with IC(50) values of 286 and 60 microM, respectively. 8. In conclusion, unlike rat and mouse pelvic ganglion neurons, which only express P2X(2) homomers, at least three distinct P2X receptors are present in guinea-pig pelvic neurons, probably homomeric P2X(2), P2X(3) and heteromeric P2X(2/3) receptors. However, some of the novel pharmacological properties observed suggest that the guinea-pig P2X receptor subtypes may differ from their rat orthologues.

Adenosine Triphosphate↗
PubMed2001

Changes in P2X receptor responses of sensory neurons from P2X3-deficient mice.

Dorsal root ganglion (DRG) neurons respond to ATP with transient, persistent or biphasic inward currents. In contrast, the ATP responses in nodose neurons are persistent. These sustained currents are also heterogeneous, with one component being accounted for by P2X2/3 receptors, and the residual response probably mediated by P2X2 receptors, although the direct evidence for this has been lacking. In the present study, we examined the P2X receptors on DRG and nodose neurons from P2X3-deficient (P2X3-/-) mice, using whole cell voltage-clamp recording and immunohistochemistry. We found that all P2X3-/- DRG neurons lacked rapidly desensitizing response to ATP, and both DRG and nodose neurons from P2X3-null mutant mice no longer responded to alpha,beta-methylene ATP (alphabetameATP). In contrast, ATP evoked persistent inward current in 12% of DRG neurons and 84% of nodose neurons from P2X3-/- mice. This retained persistent response to ATP on nodose neurons had an EC50 for ATP of 77 microm, was antagonized by Cibacron blue and pyridoxal-5-phosphate-6-azophenyl-2',4'-disulphonic acid, potentiated by Zn2+ and acidification, but not enhanced by ivermectin or diinosine pentaphosphate. 2',3'-O-Trinitrophenyl-ATP antagonized this response with an IC50 of 8 microm. All these properties are consistent with those of recombinant P2X2 homomeric receptors. Furthermore, specific P2X2 receptor immunoreactivity detected in wild-type sensory neurons was unaltered in null mutant mice. Therefore, the alphabetameATP-insensitive persistent responses on nodose neurons are likely to be mediated by P2X2 homomers, which contribute to 60% of currents evoked by 100 microm ATP in the wild type.

Adenosine Triphosphate↗
PubMed2001

Disruption of cell adhesion and caspase-mediated proteolysis of beta- and gamma-catenins and APC protein in paclitaxel-induced apoptosis.

Cell adhesion is important in the regulation of cell proliferation, migration, survival, and apoptosis. The major components of cell adhesion are the cadherin family of proteins, alpha-, beta- and gamma-catenins, and cytoskeletons. In addition, beta-catenin, when associated with adenomatous polyposis coli (APC) protein, an oncosuppressor, is implicated in the regulation of beta-catenin/APC-related signaling pathways. To examine the correlation between impairment of cell adhesion events and apoptosis, we used human non-small-cell lung cancer H460 and H520 cell lines as models to determine whether paclitaxel-induced apoptosis is associated with disruption of the components of cell adhesion and their functions. Paclitaxel treatment resulted in cells rounding up and losing contact with their neighboring cells, suggesting that the drug does indeed affect cell adhesion and related events. Western blot analysis revealed that paclitaxel caused a time- and concentration-dependent cleavage of beta-catenin, gamma-catenin, and APC protein, but not alpha-catenin or E-cadherin. These cleavages of beta-catenin and gamma-catenin were apoptosis-dependent, not mitosis-dependent. Paclitaxel treatment led to the proteolysis and activation of caspase-3 and -7, but not caspase-1. Furthermore, paclitaxel-induced apoptosis and cleavage of beta-catenin and gamma-catenin were inhibited by the pan-caspase inhibitor Z-VAD-FMK and partially inhibited by the caspase-3 inhibitor Z-DEVD-FMK but were not affected by the caspase-1 inhibitor AC-YVAD-CMK. Although the pan-caspase inhibitor blocked the cleavage of beta-catenin as well as DNA fragmentation, it did not affect paclitaxel-induced M-phase arrest and only partially prevented cell-growth inhibition. Biochemical studies revealed that cleaved beta-catenin was detected only in the Triton X-100 insoluble fraction, suggesting that it might localize in nuclear and/or membrane structures. Interestingly, the paclitaxel-induced beta-catenin fragment lost its ability to bind to E-cadherin, alpha-catenin, or APC protein and to serve as a substrate for tyrosine kinase. All our data demonstrate that the caspase-mediated cleavage of beta-catenin, gamma-catenin, and APC protein might contribute to paclitaxel-induced apoptosis.

Adenomatous Polyposis Coli Protein↗
PubMed2001

Altered SR protein expression associated with contractile dysfunction in diabetic rat hearts.

The goal of this study was to examine whether alteration of sarcoplasmic reticulum (SR) protein levels is associated with early-onset diastolic and late-onset systolic dysfunction in streptozotocin (STZ)-induced diabetic rat hearts. Four-week diabetic rat hearts exhibited slow relaxation, whereas 6-wk diabetic rat hearts exhibited slow and depressed contraction. Total phospholamban level was increased, and phosphorylated level was decreased in 4- and 6-wk diabetic rat hearts. Sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2) protein level was unchanged in 4-wk but decreased in 6-wk diabetic rat hearts. Only the apparent affinity of SR Ca2+ uptake for Ca2+ was decreased in 4-wk diabetic rat hearts, but the apparent affinity and the maximum rate was decreased in 6-wk diabetic rat hearts. Insulin treatment of the diabetic rats normalized SR protein expression and function. It was concluded that an increase in nonphosphorylated phospholamban and a decrease in the apparent affinity of SR Ca2+ pump for Ca2+ are associated with early-onset diastolic dysfunction and decreases in SERCA2 protein level and apparent affinity and maximum velocity of SR Ca2+ pump are associated with late-onset systolic dysfunction in diabetic rats.

Animals↗
PubMed2001

Loss of H-cadherin protein expression in human non-small cell lung cancer is associated with tumorigenicity.

Abnormalities in the H-cadherin gene have been described in several human cancers. However, their biological significance remains undetermined. To investigate the role of H-cadherin in non-small cell lung cancer (NSCLC), a chimera H-cadherin-green fluorescent protein (GFP) expressed in Cos-7 cells was used to identify an anti-H- cadherin antibody, HCD-1. Western blot analysis was performed in six NSCLC cell lines and 35 pairs of primary NSCLC tumors and nonmalignant lung tissue obtained from surgical resections using HCD-1. Loss of H-cadherin expression was seen in five (83%) of the six NSCLC cell lines, whereas loss of E-cadherin was seen in three (50%) of the six. H-cadherin expression was lost in 15 (43%) of 35 NSCLC surgical tumor specimens, whereas E-cadherin expression was lost in 6 (17%) of 35. H-cadherin was expressed in all of the nonmalignant lung tissue from all of the surgical specimens. Fourteen of 35 tumors were heterotransplanted s.c. in nude mice. Tumorigenicity in nude mice was associated with both loss of H-cadherin expression (P = 0.03) and loss of E-cadherin expression (P = 0.05). Loss of H-cadherin was also associated with a more advanced local tumor growth, although the difference was not significant. The results indicate that loss of H-cadherin is frequent in human NSCLC and suggest that it facilitates the implantation and local growth of human NSCLC tumors.

Animals↗
PubMed2001

[The preparation of human single chain Fv antibody specifically against hepatitis C virus NS3 antigen and its application in immunohistochemistry].

OBJECTIVE: Immunohistochemistry identification of human single chain Fv antibody (ScFv) against hepatitis C viral NS3 antigen. METHODS: The recombinant phages were panned by NS3 antigen which was coated on a microtiter plate, after five rounds of biopanning, 66 phage clones were identified specific to NS3 antigen. The affinity and specificity of ScFv were evaluated by ELISA, dot blot hybridization and immunohistochemistry. RESULTS: The data of ScFv-NS3 DNA digestion and DNA sequencing showed that the ScFv gene is composed of 747 bp. ELISA, dot blot hybridization and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C NS3 antigen has a specific combination character with hepatitis C virus NS3 antigen of different sources and paraffin-embedded patients tissues, it didn't react with liver tissue and HBsAg. CONCLUSION: The application of HCV NS3 specific ScFv in immunohistochemistry was successfully achieved.

Cells, Cultured↗
PubMed2001

[Screening and characterization of human phage antibody to hepatitis C core antigen].

OBJECTIVE: To screen and characterize human phage antibody (ScFv) against hepatitis C core antigen. METHODS: The recombinant phages were panned by core antigen that was coated in a microtiter plate. After three rounds of biopanning, 48 clones were determined specific to core antigen. The specificity of each ScFv colone was determined by ELISA. The coding gene for HCV protein ScFv has been sequenced. RESULTS: Phage antibody for HCV core protein had a specific combination character with core antigen of hepatitis C virus. The DNA sequence data showed that the ScFv gene included 774 bp. CONCLUSIONS: Human single chain antibody to hepatitis C core antigen has been identified by means of the phage display technology.

Amino Acid Sequence↗
PubMed2001

[The relation between expression of basic fibroblast growth factor and mast cells in pterygium].

OBJECTIVE: To examine the expression and distribution of basic fibroblast growth factor (bFGF) and mast cells in pterygium, and evaluate its effects in the pterygium formation and progression. METHOD: The expression of bFGF and mast cell tryptase in 17 primary pterygia, 6 recurrent pterygia and 6 normal conjunctival specimens were studied. The mast cell count and bFGF expression situation were observed. RESULTS: The bFGF was specifically localized in the epithelium, blood vessels and a subset of connective tissue cells. The bFGF expression was increased in the recurrent pterygium. The numbers of infiltrating mast cells (five 400 x sights) were (45.47 +/- 5.50) cells and (48.83 +/- 3.19) cells in the primary and recurrent pterygium respectively. In the comparisons between the cells in the pterygium (primary and recurrent) and (4.24 +/- 2.36) cells in the normal connective tissue, there were significant differences (F = 200.3128; q = 26.6762, 23.7341; P < 0.05). The shape and distribution of all the tryptase-positive cells (mast cells) in the pterygium tissues were similar to that of the cells with bFGF expression in the connective tissue. And the majority of bFGF-positive cells (87.54 +/- 3.60)% were similar to that of mast cells in the connective tissue. CONCLUSIONS: All infiltrating mast cells in pterygium have bFGF-positive expression. The bFGF expression is increased in the epithelium, blood vessels and infiltrating mast cells of the pterygium, and may contribute to the formation and progression of a pterygium.

Adult↗
PubMed2001

[Screening and characterization of human phage antibody to hepatitis virus C NS5A antigen].

OBJECTIVE: To screen and characterize human phage antibody against hepatitis C virus (HCV)NS5A antigen. METHODS: The recombinant phages were panned by recombinant HCV NS5A antigen which was coated in a microtiter plate; after five rounds of biopanning, 35 clones were obtained and demonstrated specific to NS5A antigen. The specific binding capacity of the ScFv antibody to HCV NS5A antigen was determined by ELISA. RESULTS: HCV NS5A phage antibody had a specific combination capacity with hepatitis C virus NS5A antigen. The DNA sequence data showed that the ScFv gene was composed of 789 bp and codes for a peptide of 262 amino acid residues. CONCLUSIONS: Human single chain antibody to hepatitis C virus NS5A antigen has been identified by means of the phage display technology.

Antibodies, Viral↗
PubMed2001

[The binding characteristics of insulin-MTX to insulin receptor].

OBJECTIVE: It has been reported that several kinds of tumors express increased insulin receptor and the molecules of insulin can be internalized in cells and may thence enter into the nuclei mediated by insulin receptor. In this study, we investigated the receptor binding characteristics of insulin-MTX for the possibility of using insulin as a carrier for carcinoma targeted therapy by receptor mediation. METHODS: MTX(methotrexate) was covalently linked to insulin directly. The insulin-MTX conjugate was purified by polyacrylamine agarose gel electrophoresis and analysed by high performance liquid chromatography and SDS- polyacrylamine agarose gel electrophoresis. Histologically confirmed human hepatocellular carcinoma specimens were obtained from patients at surgery and immediately frozen under -80 degrees C. Cell membrane fractions were isolated by sucrose density gradient centrifugation. Competitive displacement of 125I-insulin with insulin and insulin-MTX binding to insulin receptor were carried out and the values of IC50 and Ki were calculated so as to observe the characteristics of insulin-MTX binding to insulin receptor. RESULTS: Insulin-MTX competed as effectively as insulin with 125I-insulin for insulin receptor. The values of IC50 and Ki for insulin-MTX were 93.82 +/- 19.32 nmol/L and 91.88 +/- 16.86 nmol/L respectively, while the values of IC50 and Ki for insulin were 5.01 +/- 1.24 nmol/L and 4.85 +/- 1.12 nmol/L respectively. CONCLUSION: Insulin-MTX could bind with insulin receptor with high affinity. The result demonstrates us that there is a possibility of using insulin as a carrier for carcinoma targeted therapy by receptor mediation.

Antimetabolites, Antineoplastic↗
PubMed2001

[The effect of frozen storage for amniotic membrane ultrastructure].

OBJECTIVE: To observe the effect of frozen storage for amniotic membrane ultrastructure, and provide the experimental evidence for clinical use. METHODS: The amniotic membranes of fresh-obtained and -80 degrees C frozen storage 30 d, 60 d, 90 d and 180 d were examined with transmission electron microscope, and the ultrastructural changes were observed. RESULTS: The epithelium in the fresh-obtained amniotic membrane was observed as intact structure, abundant organelles in cytoplasm and intercellular contact with desmosome. A great quantity of collagen micro-fibrils were found in the stroma, and the light and shade striation were observed in the collagen micro-fibrils of fresh-obtained amniotic membrane. The epithelium degenerated and died gradually as the increase of frozen storage time. At the frozen storage of 90 days, the epithelial chromatin dissolved, organelles degenerated and mitochondrion vacuolated. At the frozen storage of 30 days and 60 days, the pattern of collagen micro-fibrils in stoma was similar to that in fresh-obtained amniotic membrane; At the frozen storage of 90 days, some of collagen micro-fibrils in stoma showed intumescence. At the frozen storage of 180 days, the number of collagen micro-fibrils in stoma decreased, and some of them showed intumescence and dissolvent. CONCLUSION: Frozen storage can result in amniotic membrane epithelium death and collagen micro-fibrils intumescence and dissolvent.

Adult↗
PubMed2001

[Studies on calluses induced from various explants of Cistanche deserticola].

Calluses were induced from various explants (flower, stem, bud and squama) of Cistanche deserticola Y.C.Ma and compared with each other. The result showed that the inducement rate of ovary and stem were higher than that of other explants which the inducement time was short and the characters of calluses were good. The effect of basal culture medium added hormone on callus reproduction was also studied. The result showed that B5 culture medium added 1 mg/L IBA and 2 mg/L 6-BA was optimal.

Cistanche↗
PubMed2000

Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient mice.

Extracellular ATP is implicated in numerous sensory processes ranging from the response to pain to the regulation of motility in visceral organs. The ATP receptor P2X3 is selectively expressed on small diameter sensory neurons, supporting this hypothesis. Here we show that mice deficient in P2X3 lose the rapidly desensitizing ATP-induced currents in dorsal root ganglion neurons. P2X3 deficiency also causes a reduction in the sustained ATP-induced currents in nodose ganglion neurons. P2X3-null mice have reduced pain-related behaviour in response to injection of ATP and formalin. Significantly, P2X3-null mice exhibit a marked urinary bladder hyporeflexia, characterized by decreased voiding frequency and increased bladder capacity, but normal bladder pressures. Immunohistochemical studies localize P2X3 to nerve fibres innervating the urinary bladder of wild-type mice, and show that loss of P2X3 does not alter sensory neuron innervation density. Thus, P2X3 is critical for peripheral pain responses and afferent pathways controlling urinary bladder volume reflexes. Antagonists to P2X3 may therefore have therapeutic potential in the treatment of disorders of urine storage and voiding such as overactive bladder.

Adenosine Triphosphate↗
PubMed2000

Guinea-pig sympathetic neurons express varying proportions of two distinct P2X receptors.

1. Characterization of P2X receptors on neurons of guinea-pig superior cervical ganglion (SCG) has been carried out using a whole-cell voltage-clamp technique. 2. Application of ATP and alpha,beta-methylene ATP (alphabeta-MeATP) produced fast activating inward currents, which desensitized slowly. The maximum response to alphabeta-MeATP was 36 +/- 23 % (range 0.1-100 %) of that evoked by ATP in the same cell. 3. Co-application of alphabeta-MeATP (300 microM) with ATP (300 microM) produced a response that was 97 +/- 1 % of that given by ATP alone. Following desensitization with alphabeta-MeATP, the decrease in response to ATP was equal to the absolute reduction in response to alphabeta-MeATP in the same cell. 4. The concentration-response curve for alphabeta-MeATP had an EC50 of 42 microM and a Hill coefficient of 1.17. For cells where the ratio of alphabeta-MeATP/ATP currents at 100 microM was < 0.1, the ATP concentration-response curve had an EC50 of 56 microM and a Hill coefficient of 1.95. However, in cells where the ratio was > 0.7, the curve had an EC50 of 60 microM and a Hill coefficient of 0.97. 5. The response to 100 microM alphabeta-MeATP was inhibited by 2' (or 3')-O-trinitrophenyl-ATP (TNP-ATP) with an IC50 of 70 nM. However, on cells where the ratio of alphabeta-MeATP/ATP currents was < 0.1, ATP was inhibited by TNP-ATP with an IC50 of 522 nM. 6. Immunohistochemical staining with antibodies raised against rat P2X2 and P2X3 epitopes suggested that both subunits were expressed by guinea-pig SCG neurons. 7. We conclude that varying proportions of two distinct P2X receptors coexist on the cell bodies of individual guinea-pig SCG neurons, which may correspond to homomeric P2X2 and heteromeric P2X2/3 receptors.

Adenosine Triphosphate↗
PubMed2000

A neurofibromatosis-1-regulated pathway is required for learning in Drosophila.

The tumour-suppressor gene Neurofibromatosis 1 (Nf1) encodes a Ras-specific GTPase activating protein (Ras-GAP). In addition to being involved in tumour formation, NF1 has been reported to cause learning defects in humans and Nf1 knockout mice. However, it remains to be determined whether the observed learning defect is secondary to abnormal development. The Drosophila NF1 protein is highly conserved, showing 60% identity of its 2,803 amino acids with human NF1 (ref. 12). Previous studies have suggested that Drosophila NF1 acts not only as a Ras-GAP but also as a possible regulator of the cAMP pathway that involves the rutabaga (rut)-encoded adenylyl cyclase. Because rut was isolated as a learning and short-term memory mutant, we have pursued the hypothesis that NF1 may affect learning through its control of the Rut-adenylyl cyclase/cAMP pathway. Here we show that NF1 affects learning and short-term memory independently of its developmental effects. We show that G-protein-activated adenylyl cyclase activity consists of NF1-independent and NF1-dependent components, and that the mechanism of the NF1-dependent activation of the Rut-adenylyl cyclase pathway is essential for mediating Drosophila learning and memory.

Adenylyl Cyclases↗