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Biomedical subjects

D Xin

Publications and source records attributed to D Xin.

At least 19 recordsLinked to original sources
PubMed2001

Morphology and physiology of the polyaxonal amacrine cells in the rabbit retina.

We examined the morphology and physiological response properties of the axon-bearing, long-range amacrine cells in the rabbit retina. These so-called polyaxonal amacrine cells all displayed two distinct systems of processes: (1) a dendritic field composed of highly branched and relatively thick processes and (2) a more extended, often sparsely branched axonal arbor derived from multiple thin axons emitted from the soma or dendritic branches. However, we distinguished six morphological types of polyaxonal cells based on differences in the fine details of their soma/dendritic/axonal architecture, level of stratification within the inner plexiform layer (IPL), and tracer coupling patterns. These morphological types also showed clear differences in their light-evoked response activity. Three of the polyaxonal amacrine cell types showed on-off responses, whereas the remaining cells showed on-center responses; we did not encounter polyaxonal cells with off-center physiology. Polyaxonal cells respected the on/off sublamination scheme in that on-off cells maintained dendritic/axonal processes in both sublamina a and b of the IPL, whereas processes of on-center cells were restricted to sublamina b. All polyaxonal amacrine cell types displayed large somatic action potentials, but we found no evidence for low-amplitude dendritic spikes that have been reported for other classes of amacrine cell. The center-receptive fields of the polyaxonal cells were comparable to the diameter of their respective dendritic arbors and, thus, were significantly smaller than their extensive axonal fields. This correspondence between receptive and dendritic field size was seen even for cells showing extensive homotypic and/or heterotypic tracer coupling to neighboring neurons. These data suggest that all polyaxonal amacrine cells are polarized functionally into receptive dendritic and transmitting axonal zones.

Amacrine Cells↗
PubMed2001

[Frequent somatic mutations of the von Hippel-Lindau tumor suppressor gene in primary sporadic human renal clear cell carcinomas].

OBJECTIVE: To investigate the mutation of Von Hippel-Lndau (VHL) tumor suppressor gene in patients with primary sporadic human renal cell carcinoma (RCC). METHODS: DNA samples from 20 primary sporadic renal clear carcinoma patients were analyzed by polymerase chain reaction, single strand conformational polymorphism analyses (PCR-SSCP) and direct sequencing. RESULTS: Somatic mutations of the VHL gene were detected in 11 (55%) of the 20 clear cell renal carcinomas, including 6 deletions, 2 insertions, and 3 missense mutations. These mutations mainly occurred in the last one-third region of exon 1, 2 and 3 of the VHL gene. CONCLUSION: The VHL gene may frequently mutate in Chinese patients with primary sporadic renal clear cell carcinoma. The VHL gene may be useful as a marker gene for the diagnosis of RCC and as a target gene for molecular therapy.

Adult↗
PubMed2001

Expression of beta-catenin in renal cell carcinoma.

OBJECTIVE: To investigate the expression of beta-catenin and its mRNA in renal cell carcinoma. METHODS: Twenty-six cases with renal cell carcinoma (RCC) were studied by immunohistochemistry, Western blot and RT-PCR. RESULTS: We found the expression of beta-catenis is higher in cancer tissues than in normal kidney tissues and the level of beta-catenin is associated with the tumor stage. Its expression in tumor of pT3 and pT4 is obviously higher than pT1 and pT2 (P < 0.01). That is to say, there was an overexpression of beta-catenin protein in RCC and its level was related to the tumor stage, but the expression of beta-catenin mRNA had no difference between tumor tissue and normal tissue. CONCLUSION: beta-catenin may be related to the occurrence and progress of RCC.

Adult↗
PubMed2001

Suppression subtractive hybridization for identifying differentially expressed genes in renal cell carcinoma.

OBJECTIVE: To construct a renal cell carcinoma (RCC) cDNA subtractive library using suppression subtractive hybridization. METHODS: Polyadenylated RNA [Poly (A)+ RNA] was isolated from tissues of RCC and normal kidney, and single-strand cDNAs and double-strand cDNAs were synthesized in turn. RCC cDNAs were divided into two groups and ligated to the specific adaptors I and 2, and then hybridized with normal kidney cDNA twice with two rounds of suppression PCR. Second round PCR products were cloned to T/A plasmid vectors to set up the subtractive library. One hundred clones were randomly picked to perform enzyme digest analysis, and some underwent sequence analysis and Northern blot to identify RCC specifically expressed genes. SMART RACE procedure was operated to clone full length novel RCC specifically expressed genes. RESULTS: A human RCC subtractive library with high subtractive efficiency was successfully set up. The amplified library contains 350 positive clones. Random analysis of 100 clones with enzyme restriction showed that 85 plasmids in the clones contained 50-400 bp inserts. Sequence analysis was performed for 10 clones. All the 10 sequences were unknown before and derived from 6 unique, novel genes among which the cDNA insert RCC18 had five copies. Northern blot analysis showed that RCC18 cDNA was highly expressed in RCC, but no signal could be detected in normal kidney. Using SMART RACE technique, we obtained the full length of the novel gene RCC18. CONCLUSIONS: The constructed cDNA subtractive library of human RCC is a highly efficient one and lays a solid foundation for large scale screening and cloning new and specific oncogenes or tumor suppressor genes of RCC. The novel specifically expressed genes provided an important clue for studying the mechanisms of occurrence and development of RCC.

Carcinoma, Renal Cell↗
PubMed2000

Surround inhibition of mammalian AII amacrine cells is generated in the proximal retina.

1. Intracellular recordings were obtained from neurons in the superfused retina-eyecup preparation of the rabbit under dark-adapted conditions. Neurotransmitter agonists and antagonists were applied exogenously via the superfusate to dissect the synaptic pathways pharmacologically and thereby determine those pathways responsible for the generation of the on-centre/off-surround receptive fields of AII amacrine cells. 2. Application of the metabotropic glutamate receptor agonist, APB, reversibly blocked both the on-centre and off-surround responses of AII cells. These data were consistent with the idea that both the centre- and surround-mediated responses are derived from inputs from the presynaptic rod bipolar cells. 3. Whereas rod bipolar cells showed on-receptive fields approximately 100 microm across, we found no evidence for an antagonistic off-surround response using light stimuli which effectively elicited the off-surrounds of AII amacrine cells. These results indicated that the surrounds of AII cells are not derived from rod bipolar cell inputs. 4. Application of the ionotropic glutamate receptor antagonists CNQX or DNQX enhanced the on-centre responses of AII cells but attenuated the off-surround responses. These data indicated that the centre- and surround-mediated responses could not both be derived from signals crossing the rod bipolar-to-AII cell synapse. 5. Application of the glycine antagonist, strychnine, had only minor and variable effects on AII cell responses. However, the GABA antagonists picrotoxin and bicuculline enhanced the on-centre response but attenuated or completely blocked the off-surround response of AII cells. The GABA antagonists had no effect on the responses of horizontal cells indicating that their effects on AII cell responses reflected actions on inner retinal circuitry rather than feedback circuitry in the outer plexiform layer. 6. Application of the voltage-gated sodium channel blocker TTX enhanced the on-centre responses of AII cells but attenuated or abolished their off-surround responses. 7. Taken together, our results suggest that the on-centre responses of AII cells result from the major excitatory drive from rod bipolar cells. However, the surround receptive fields of AII cells appear to be generated by lateral, inhibitory signals derived from neighbouring GABAergic, on-centre amacrine cells. A model is presented whereby the S1 amacrine cells produce the surround receptive fields of AII amacrine cells via inhibitory, feedback circuitry to the axon terminals of rod bipolar cells.

Aminobutyrates↗
PubMed2000

Different Wnt-5A gene expressions in the renal cell carcinoma GRC-1 cell line during the cell cycle.

OBJECTIVE: To investigate the gene expression at transcription level of growth factor Wnt-5A in different phase during the cell cycle. METHODS: We synchronized the renal cell carcinoma GRC-1 cell line by double thymidine blocks and high-pressure N2O gae methods and amplified Wnt-5A cDNAs from different phase using Semi-quantitative RT-PCR (reverse transcriptase polymerase chain reaction). The PCR products were electrophoresized on the agrose gel and detected by Gel Doc 1000 computer controlled system integrating the volumes of each band, representing the intensities of all pixels in a defined band. RESULTS: The different mRNA expressions of growth factor Wnt-5A was detected in RCC GRC-1 cell line. In S phase, the highest level of Wnt-5A transcript was observed, and in G1 and M phase, medial and lowest, respectively. The differences between S and M stages were statistically significant (P < 0.05). CONCLUSION: Growth factor Wnt-5A has the potential effect on tumorigenesis. It contributes to all phases during cell cycle but in S phase especially.

Carcinoma, Renal Cell↗
PubMed2000

[Beta-catenin and its mRNA in renal cell carcinoma].

OBJECTIVES: To investigate the role of beta-catenin in renal cell carcinoma. METHODS: The expression of beta-catenin in 26 cases of renal cell carcinoma was studied by LSAB immunohistochemistry, Western blotting and RT-PCR. RESULTS: The expression of beta-catenin was higher in cancer tissues than in normal kidney tissues in 25 cases and the level of beta-catenin was associated with tumor stage. The expression was obviously higher in pT3 and pT4 than in pT1 and pT2 (P < 0.01). The expression of its mRNA was not higher in tumor cells than in normal cells. Beta-catenin was expressed mainly in cytoplasm. CONCLUSIONS: Beta-catenin might be related to the occurrence and development of kidney tumor.

Adult↗
PubMed2000

[Cloning and identifying of renal cell carcinoma differentially expressed genes].

OBJECTIVE: To clone and identify RCC specially expressed genes different with normal kidney tissue. METHODS: A technique called suppression subtractive hybridization was used to construct the library which contains the differently expressing cDNAs between RCC and normal kidney. Then the RCC specially expressed genes were cloned from it. RESULTS: Human RCC subtractive library with high subtractive efficiency was set up successfully. The amplified library contains 350 positive clones. Sequence analysis was performed in 5 clones. All of the sequences were unknown before and the cDNA inserting GYLZ-RCC18 had three copies. Northern blot analysis showed that GYLZ-RCC18 cDNA expressed highly in RCC, but there was no any signal could be detected in normal kidney. Using SMART RACE technique, we obtained the full length of novel gene of GYLZ-RCC18. We also identified that GYLZ-RCC18 family contains 3 subtype genes. CONCLUSIONS: The highly efficient cDNA subtractive library lays a solid foundation for screening and cloning new and specific oncogenes or tumor suppressor genes of RCC. The novel differentially expressed genes provide an important clue to studying the mechanism of the occurrence and development of RCC.

Carcinoma, Renal Cell↗
PubMed1999

Dark- and light-induced changes in coupling between horizontal cells in mammalian retina.

Retinal horizontal cells exhibit large receptive fields derived from their extensive electrical coupling by means of gap junctions. The conductance of these gap junctions seems to be regulated by dopamine acting through a cAMP-mediated cascade. There is now abundant evidence that extracellular dopamine levels vary with changes in ambient light intensity, suggesting that changes in the dark/light adaptational state of the retina can modulate coupling between horizontal cells. We studied this question in the mammalian retina by determining the effects of ambient light levels, in the form of changing background light intensity, on the coupling profiles of A- and B-type horizontal cells in the rabbit. Changes in coupling were assessed by measurements of the space constants of the syncytium formed by horizontal cells and the intercellular spread of the biotinylated tracer Neurobiotin. Our results indicate that dark-adapted horizontal cells show relatively weak coupling. However, presentation of background lights as dim as one-quarter log unit above rod threshold resulted in increases in both the averaged extent of tracer coupling and space constants of A- and B-type horizontal cells. Coupling expanded further as background light intensities were increased by 1-1.5 log units, after which additional light adaptation brought about an uncoupling of cells. Coupling reached its minimum at light intensities about 3 log units above rod threshold, after which, with further light adaptation, it stabilized at levels close to those seen in dark-adapted retinas. Our results indicate that electrical coupling between mammalian horizontal cells is modulated dramatically by changes in the adaptational state of the retina: coupling is maximized under dim ambient light conditions and diminishes as the retina is dark or light adapted from this level.

Adaptation, Ocular↗
PubMed1999

Effect of verbascoside on decreasing concentration of oxygen free radicals and lipid peroxidation in skeletal muscle.

AIM: To detect the effects of verbascoside on decreasing the concentration of oxygen free radicals (OFR) and lipid peroxidation in skeletal muscle resulting from exhaustive exercise. METHODS: Electron spin resonance (ESR) technique and thiobarbituric acid reaction (TBAR) method were used to detect the concentration of OFR in intact gastrocnemius muscle and the contents of milondialdehyde (MDA) in muscle homogenate. RESULTS: Verbascoside decreased the concentration of OFR (P < 0.05) and the level of lipid peroxidation (P < 0.05) in muscle caused by exercise. CONCLUSION: Verbascoside has the effects of reducing oxidative stress in muscle caused by exhaustive exercise by decreasing the concentration of free radicals and the level of lipid peroxidation.

Animals↗
PubMed1999

[Investigation of factors affecting the development of COPD].

OBJECTIVE: To investigate the factors which affect the development of COPD. METHODS: 88 patients with mPAP less than 20 mm Hg were followed up by 5 - 20 years. Pulmonary ventilation, blood gas analysis, ECG, pulmonary hemodynamic parameters and oxygen saturation at night were performed one time every 5 years. RESULTS: The results showed that: 5 year, 10 year and 20 year's mortality is 6%, 25% and 43% respectively; MVV%, FEV(1)% and PaO(2) declined accompanied by the increase of PaCO(2); among 41 patients with mPAP 16 - 20 mm Hg, 20 patients' mPAP were more than 20 mm Hg and 15 patients died after 10 years' follow up. Among 16 patients with the declination of oxygen saturation, 8 patients' mPAP were more than 20 mm Hg and 6 patients died. 57 strains of infectious bacteria were found in 88 patients' sputum with 43 strains in the death group. CONCLUSIONS: These results demonstrated that COPD is a kind of chronic diseases with high death rate; infection, decline of oxygen saturation at night and mPAP more than 16 mm Hg were high risk factors in COPD.

Adult↗
PubMed1998

Mitochondrial DNA in platelets from aged subjects.

This study aimed to assess platelets as a possible model for screening the accumulation of mitochondrial DNA mutations, particularly during normal ageing. For this purpose we isolated platelets from young and old donors selected by lack of systemic and haematological diseases. We studied the accumulation of a particular deletion (4977-bp deletion) that usually accumulates in an age-related manner in different post-mitotic tissues, such as brain, heart and skeletal muscle, and in some non-post-mitotic tissues (skin, liver). Using different primers, we failed to detect this particular species of deletion in platelets both from young and old individuals. However, we cannot exclude the presence of other species of deletions or point mutations affecting the mitochondrial DNA in platelets during the aging process.

Adolescent↗
PubMed1998

Cloning and sequencing of the rat cDNAs encoding class I beta-tubulin.

Two kinds of rat cDNA clones encoding class I beta-tubulin were isolated from the neonatal rat brain, and sequenced. They corresponded to mRNA species formed by the alternative usage of polyadenylation signals. The determined nucleotide sequence showed high (84.6%) identity to rat class II beta-tubulin, the only rat beta-tubulin sequence reported in the nucleotide database, in the coding region, but relatively low (47.3-50.7%) in the noncoding region. The Northern blot analysis using the 3'-noncoding fragment as a probe showed that both mRNA species of the class I beta-tubulin were down-regulated during brain development.

Amino Acid Sequence↗
PubMed1997

Tracer coupling pattern of amacrine and ganglion cells in the rabbit retina.

We examined the tracer coupling pattern of more than 15 morphological types of amacrine and ganglion cells in the rabbit retina. Individual cells were injected intracellularly with the biotinylated tracer Neurobiotin, which was then allowed to diffuse across gap junctions to label neighboring neurons. We found that homologous and/or heterologous tracer coupling was common for most proximal neurons. In fact, the starburst amacrine cell was the only amacrine cell type that showed no evidence of coupling. The remaining types of amacrine cell were coupled exclusively to other amacrines, either homologously or, more often, through a combination of homologous and heterologous junctions. In only one case did we visualize labeled ganglion cells following injection of Neurobiotin into an amacrine cell. In contrast, injection of Neurobiotin into ganglion cells almost always resulted in the labeling of amacrine cells. Taken together, these results suggest a directionality to the movement of tracer across gap junctions connecting amacrine and ganglion cells. We found that the coupling pattern for a given morphological type of cell was generally stereotypic and consistent across retinas. The notable exceptions to this finding were alpha ganglion cells and cells with morphology corresponding to that of on-off direction selective ganglion cells. In both cases, individual cells showed either extensive coupling to both amacrine and ganglion cells or no coupling at all. A notable finding was that, in every case, the neighboring cells within a tracer-coupled array were always within one gap junction of the injected neuron. Furthermore, in many cases, the array formed by the somata of tracer-coupled cells was almost perfectly coincident with the dendritic arbor of the injected cell. Thus, our results indicate that whereas coupling is extensive within the proximal retina, individual cells partake in coupled networks that are stereotypic and highly circumscribed.

Animals↗
PubMed1997

[Role of pulmonary arterial angiotensin converting enzyme in development of hypoxic pulmonary hypertension].

OBJECTIVE: To study the effect of local pulmonary arterial renin-angiotensin system on hypoxic pulmonary hypertension(PAH). METHOD: Normobaric hypoxic PAH animal model was adopted. Systolic pulmonary arterial pressure(PAPs), pulmonary arterial angiotensin converting enzyme(ACE) activity, collagen content(HP) were measured and ultrastructure of pulmonary smooth muscle cells (SMC) and was observed. RESULTS: During hypoxia, PAPs, pulmonary ACE activity, HP increased significantly and SMC changed into secretive phenotype. Treatment with cilazapril, an ACE inhibitor, decreased ACE activity along with reversal of PAPs. HP and SMC phenotype. CONCLUSIONS: Pulmonary ACE participated in the modulation of changes of SMC phenotype and collagen metabolism during hypoxic PAH.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1996

[The significance of circulating endothelial cell in hypoxic injury of pulmonary vascular endothelial cell].

OBJECTIVE: To observe the changes of vascular endothelial cell (VEC) in hypoxic rats and to check the damage of VEC in vivo, an experiment for observing circulating endothelial cells was conducted. METHOD: Thirty male Wistar rats were classified into 5 groups in random, exposted to 10% O2 and 1% FeCl3 intraveniously. After 0, 1, 2, 3, 4 weeks, PaO2, circulating endothelial cells (CEC) were measured and the lung specimens, the pulmonary artery were examined by transmission electron micrography (TEM) and scanning electron micrography (SEM). RESULTS: Lung specimens and pulmonary artery under TEM and SEM: swollen appeared to VEC after 1 week. As time prolonged, VEC appeared splitting, broken and peeled off. The amount of CEC increased as the time proceeding, the longer the time, the higher the number of the CEC. Obvious hypoxemia developed in the tested animals as the time prolonged. CONCLUSIONS: All above results indicated: the time of anoxia proceeding, the structure of lung and pulmonary artery VEC were damaged obviously. The increase of CEC matched with the changes of ultrastructure of lung VEC. CEC specifically reflected the damage of VEC. Non-invasive measurement of CEC is valuble in evaluation of hypoxic injury of pulmonary VEC.

Animals↗
PubMed1994

Visual evoked potentials following abrupt contrast changes.

The timing of visual evoked potential (VEP) amplitude and phase changes following abrupt increases or decreases in contrast was examined. Gratings (1 c/deg) were presented at a low contrast for 8 sec, increased to a higher contrast for 8 sec, and then decreased to the initial lower contrast for another 8 sec. Second harmonic VEP amplitude and phase were recorded continuously and averaged in 1 sec epochs. Both amplitude and phase exhibited delays in reaching a stable level following the contrast change. For amplitude, the length of the delay was dependent on the magnitude and direction of the contrast step and on the spatial frequency of the stimulus. Time constants for the change in amplitude following step increases in contrast ranged from 0.2 sec for a 12% contrast step to 1.34 sec for a 37% contrast step. The timing of phase changes, however, was independent of the size of the contrast increases (tau = 0.7 sec). For step decreases in contrast, both amplitude and phase were relatively independent of the size of the change (tau = approx. 0.9 sec for amplitude and tau = 0.15 sec for phase). Amplitude time constants also increased with increasing spatial frequency (tau = 1.2 sec for 1 c/deg, tau = 1.6 sec for 4 c/deg and tau = 2.3 sec for 8 c/deg); phase time constants, however, did not change as a function of spatial frequency (tau = 0.7 for all spatial frequencies). These findings demonstrate that a unitary process may not always be tapped by signal averaging techniques. Additionally, swept stimulus VEP techniques may produce considerable errors in threshold estimation depending on the stimulus spatial frequency and on the slope and direction of the contrast change.

Contrast Sensitivity↗