Biomedical subjects

W Jin

Publications and source records attributed to W Jin.

At least 73 records · Page 4Linked to original sources

PubMed2000

Mitochondrial gene variation in type 2 diabetes mellitus: detection of a novel mutation associated with maternally inherited diabetes in a Chinese family.

OBJECTIVE: To explore the relationship between type 2 diabetes mellitus and the mutation(s) in mitochondrial DNA. METHODS: According to the previous literature, the fragment of mitochondrial DNA from nucleotide 3153 to 3551, which had shown high frequency of point mutation, was scanned with the technique of polymerase chain reaction--single strand conformation polymorphism (PCR-SSCP) in Chinese normal control, type 2 diabetic population, and 12 families suffered from maternally inherited type 2 diabetes mellitus. Direct sequencing was applied to detect the fragments with abnormal conformation. RESULTS: No special band was found in SSCP electrophoreses in Chinese normal control, and only one subject (No. 81) of diabetic population indicated the abnormality in SSCP study, which was affirmed to be a silent point mutation of T to C at nucleotide 3336 inducing no change in amino acid (ATT-->ATC, Ile). Pedigree 25,001 was the only family that exhibited strongly different SSCP characteristic from the other 11 ones, which was confirmed to be caused by a single point mutation mt3285T-->C/T in the coding region of tRNA(Leu(UUR)) gene by the technique of direct sequencing. CONCLUSIONS: The variation within mt DNA 3153-3551 is not the major cause of type 2 diabetes in Chinese population suffered from this disease in this study. The point mutation T-->C/T at the mitochondrial nucleotide 3285, which was found in pedigree 25,001, is located in the highly conservative region of tRNA(Leu(UUR)) gene. It is strongly suggested that this mutation cause the conversion in the 3-dimentional structure of tRNA(Leu(UUR)), which might disturb the normal translation and lead to the impairment in mitochondrial oxidative phosphorylation characterized by the defects of the polypeptides involved in the respiratory chain. Thus, insulin secretion deficiency and insulin resistance might occur.

Adult↗

PubMed2000

p53 mutation, EGFR gene amplification and loss of heterozygosity on chromosome 10, 17 p in human gliomas.

OBJECTIVE: To further illustrate the roles of p53 gene, epidermal growth factor receptor (EGFR) gene and loss of heterozygosity (LOH) on chromosome 10 and 17 p in human glioma progression. METHODS: p53 mutations were scanned in 50 gliomas with various malignant grades using the polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) assay, and were confirmed by direct sequencing. LOH for chromosome 10, 17 p and amplification of the EGFR gene were also assessed using Southern blot analysis. RESULTS: p53 mutations were found in 9 of 17 high-grade astrocytomas (53%), 1 of 15 low-grade astrocytomas (7%), and the only subject of eppendymoblastoma but in none of the 10 medulloblastomas and 7 eppendymomas. The majority of gliomas (38/50) analyzed here retained both 17 p alleles. The frequency of p53 mutations was 13% in this group of tumors and increased to 50% (6/12) in tumors with one 17 p allele (P < 0.025). LOH on chromosome 10 was found in 35% (6/17) of high-grade astrocytomas, in 10% (1/10) of medulloblastomas, but in 0% of low-grade gliomas. EGFR gene amplification was found in 9 high-grade gliomas, 60% (6/9) of which also presented LOH for chromosome 10. CONCLUSIONS: These results indicate that p53 inactivation is a common genetic event in astrocytoma progression that may be more strongly associated with the progression of astrocytomas than with their origin. Absence of p53 mutations in 50% of the tumors with one 17 p allele suggests that a tumor suppressor gene other than p53 may be located on chromosome 17 p and involved in progression to malignancy of some gliomas. The loss of alleles on chromosome 10 and the amplification of the EGFR gene appear to be restricted to high-grade tumors, suggesting that these events may be related to tumor progression rather than initiation.

Brain Neoplasms↗

PubMed2000

[Studies on vaccination schedule of inactivated hepatitis A vaccine].

OBJECTIVE: To evaluate three vaccination schedules of inactivated hepatitis A vaccine in rhesus monkeys (Macaca Mulatta). METHODS: Seventeen rhesus monkeys were divided randomly into four groups. The first group of five monkeys was immunized with vaccination schedule 1 (one immunization at week 0); The second group of five monkeys was immunized with vaccination schedule 2 (first injection at week 0, and booster one at week 4); the third group of five monkeys was immunized with vaccination schedule 3 (first injection at week 0, and booster one at week 24); and the fourth group of two monkeys with MEM as control. Rhesus monkeys were inoculated by intramuscular injections to detect the ALT (alanine aminotransferase), liver pathology and anti-HAV at different time. RESULTS: During the 52-week observation, no abnormal elevation of ALT or liver pathology appeared. The IgG response for vaccination schedule 1 reached the highest level (13 219 mIU/ml), and decreased gradually to 2 056 mIU/ml at week 52. After booster immunization at week 4, the IgG response for vaccination schedule 2 was 13 863 mIU/ml at week 12, and decreased to 6 285 mIU/ml. The highest level of IgG (40 638 mIU/ml) for vaccination schedule 3 appeared at week 28 after booster injection at week 24. Further more, at week 52 the IgG level for vaccination schedule 3 was also the highest among the three vaccination schedules. CONCLUSIONS: The inactivated hepatitis A vaccine is safe and well immunogenic. The vaccination schedule 3 showed the best immune response among the three vaccination schedules.

Adult↗

PubMed2000

[Effect of low dose gossypol treatment on male sperm nuclear basic protein].

OBJECTIVE: Study on human sperm nuclear basic proteins after low dose of gossypol treatment. METHODS: Twenty-three men with normal fertility were divided into two groups. The experimental group included 15 men which were treated daily with gossypol (15 mg/d for 12 weeks and treated with 10 mg/d for 32 weeks continuously), 8 men were used in the control group. Semen was collected at 1, 4, 8, 12, 16, 24, 36 and 44 weeks after gossypol treatment and 10 weeks after stopping treatment. Total nuclear basic protein (TNBP) was extracted and TH/TP, HP1/(HP2+HP3) ratios were determined by scanning microdensitometry following electrophoresis of TNBP in polyacrylamide gels. RESULTS: The TH/TP ratio of control and the gossypol treated group were 0.17 +/- 0.07 and 0.63 +/- 0.79(P < 0.05); The HP1/(HP2+HP3) ratio of control and gossypol treated group were 1.05 +/- 0.21 and 1.47 +/- 0.18 (P < 0.01). These two parameters were returned to normal after 10 weeks of stopping gossypol treatment. CONCLUSIONS: The interruption of histone-to-protamine replacement reaction (HPRR) and alteration of nuclear basic proteins induced by low dose gossypol treatment might lead to infertility. Changes of HPRR and nuclear basic protein were reversible after stopping gossypol treatment.

Adult↗

PubMed1999

Synthesis of a stable form of tertiapin: a high-affinity inhibitor for inward-rectifier K+ channels.

Tertiapin (TPN), a small protein derived from honey bee venom, inhibits the GIRK1/4 and ROMK1 channels with nanomolar affinities. Methionine residue 13 in TPN interacts with residue F148 in the channel, located just outside of the narrow region of the ROMK1 pore. The methionine residue in TPN can be oxidized by air, which significantly hinders TPN binding to the channels. To overcome the reduction in TPN affinity due to oxidation of M13, we replaced M13 in TPN with fourteen different residues. Out of the fourteen derivatives, only the one in which M13 was replaced by glutamine, TPNQ, binds to the channel with a Ki value very similar to that of native TPN. Since TPNQ is stable and functionally resembles native TPN, it will be a very useful molecular probe for studying the inward-rectifier K+ channels.

Animals↗

PubMed1999

Mechanisms of inward-rectifier K+ channel inhibition by tertiapin-Q.

Tertiapin-Q (TPN(Q)) is a derivative of honey bee toxin tertiapin (TPN) whose methionine residue is replaced with a glutamine residue. TPN(Q) inhibits the ROMK1 and GIRK1/4 inward-rectifier K(+) channels with affinities very similar to TPN. However, unlike native TPN, TPN(Q) is nonoxidizable by air. The stability of TPN(Q) allows us to investigate how it interacts with the targeted channels. We found that the interaction between TPN(Q) and the ROMK1 channel is a bimolecular reaction, i.e., one TPN(Q) molecule binds to one channel. The interaction surface in TPN(Q) is primarily formed by its alpha helix rather than the beta sheets with which scorpion toxins form their interaction surface. The mutagenesis studies on both the channel and TPN(Q) together strongly suggest that to block the K(+) pore TPN(Q) plugs its alpha helix into the vestibule of the K(+) pore, while leaving the extended structural portion sticking out of the vestibule into the extracellular media.

Alanine↗

PubMed1999

Redundant intronic repressors function to inhibit fibroblast growth factor receptor-1 alpha-exon recognition in glioblastoma cells.

The human fibroblast growth factor receptor-1 primary transcript is alternatively processed to produce receptor forms that vary in their affinity for fibroblast growth factor. The inclusion of a single exon (alpha) in normal brain glial cells produces a low affinity form of the receptor. Recognition of the alpha-exon is dysregulated during neoplastic transformation of glial cells to produce a high affinity receptor form. In this study, we have identified a second intronic repressor of RNA splicing located approximately 250 nucleotides upstream of the alpha-exon. Deletion or mutation of this sequence resulted in a significant increase in exon recognition in glioblastoma cells. This intronic repressor was found to share significant sequence homology with an intronic repressor element located downstream of the alpha-exon. The two repressor elements are functionally redundant in that they are capable of inhibiting alpha-exon recognition when positioned upstream or downstream of the exon. Finally, the elements were found to mediate enhanced exclusion of an unrelated exon, but only the repressors were placed flanking the exon. However, under these conditions, the cell-specific exon exclusion was no longer maintained. These results suggest that although the alpha-exon inclusion is actively repressed in glioblastomas, the absence of trans-activators appears to be key to the production of the high affinity form of fibroblast growth factor receptor-1 in glioblastomas.

Alternative Splicing↗

PubMed1999

Secretory leukocyte protease inhibitor suppresses the inflammation and joint damage of bacterial cell wall-induced arthritis.

Disruption of the balance between proteases and protease inhibitors is often associated with pathologic tissue destruction. To explore the therapeutic potential of secretory leukocyte protease inhibitor (SLPI) in erosive joint diseases, we cloned, sequenced, and expressed active rat SLPI, which shares the protease-reactive site found in human SLPI. In a rat streptococcal cell wall (SCW)-induced model of inflammatory erosive polyarthritis, endogenous SLPI was unexpectedly upregulated at both mRNA and protein levels in inflamed joint tissues. Systemic delivery of purified recombinant rat SLPI inhibited joint inflammation and cartilage and bone destruction. Inflammatory pathways as reflected by circulating tumor necrosis factor alpha and nuclear factor kappaB activation and cartilage resorption detected by circulating levels of type II collagen collagenase-generated cleavage products were all diminished by SLPI treatment in acute and chronic arthritis, indicating that the action of SLPI may extend beyond inhibition of serine proteases.

Amino Acid Sequence↗

PubMed1999

Distinct domains of the CB1 cannabinoid receptor mediate desensitization and internalization.

Desensitization of cannabinoid receptor signaling by a G-protein coupled receptor kinase (GRK) was examined using the Xenopus oocyte expression system. Application of a CB1 agonist, WIN 55,212-2, evoked a concentration-dependent increase in K+ conductance (Kir3) in oocytes coexpressing rat CB1 with the G-protein-gated, inwardly rectifying K+ channels Kir3.1 and Kir3.4. Desensitization was slight during continuous agonist application in the absence of GRK and arrestin. However, coexpression of GRK3 and beta-arrestin 2 (beta-arr2) caused profound homologous CB1 receptor desensitization, supporting the hypothesis that GRK3 and beta-arr2 effectively produce CB1 receptor desensitization. To identify the regions of the CB1 receptor responsible for GRK3- and beta-arr2-mediated desensitization, we constructed several CB1 receptor mutants. Truncation of the C-terminal tail of CB1 receptor at residue 418 (Delta418) almost completely abolished desensitization but did not affect agonist activation of Kir3. In contrast, truncation at residues 439 and 460 did not significantly affect GRK3- and beta-arr2-dependent desensitization. A deletion mutant (Delta418-439) did not desensitize, indicating that residues within this region are important for GRK3- and beta-arr2-mediated desensitization. Phosphorylation in this region was likely involved in desensitization, because mutation of either of two putative phosphorylation sites (S426A or S430A) significantly attenuated desensitization. CB1 receptors rapidly internalize after activation by agonist. Phosphorylation of S426 or S430 was not necessary for internalization, because the S426A/S430A CB1 mutant internalized when stably expressed in AtT20 cells. These studies establish that CB1 desensitization can be regulated by a GRK and that different receptor domains are involved in GRK- and beta-arrestin-dependent desensitization and CB1 internalization.

Amino Acid Sequence↗

PubMed1999

Characterization and expression of the laminin gamma3 chain: a novel, non-basement membrane-associated, laminin chain.

Laminins are heterotrimeric molecules composed of an alpha, a beta, and a gamma chain; they have broad functional roles in development and in stabilizing epithelial structures. Here, we identified a novel laminin, composed of known alpha and beta chains but containing a novel gamma chain, gamma3. We have cloned gene encoding this chain, LAMC3, which maps to chromosome 9 at q31-34. Protein and cDNA analyses demonstrate that gamma3 contains all the expected domains of a gamma chain, including two consensus glycosylation sites and a putative nidogen-binding site. This suggests that gamma3-containing laminins are likely to exist in a stable matrix. Studies of the tissue distribution of gamma3 chain show that it is broadly expressed in: skin, heart, lung, and the reproductive tracts. In skin, gamma3 protein is seen within the basement membrane of the dermal-epidermal junction at points of nerve penetration. The gamma3 chain is also a prominent element of the apical surface of ciliated epithelial cells of: lung, oviduct, epididymis, ductus deferens, and seminiferous tubules. The distribution of gamma3-containing laminins on the apical surfaces of a variety of epithelial tissues is novel and suggests that they are not found within ultrastructurally defined basement membranes. It seems likely that these apical laminins are important in the morphogenesis and structural stability of the ciliated processes of these cells.

Amino Acid Sequence↗

PubMed1999

Mu opioids enhance mossy fiber synaptic transmission indirectly by reducing GABAB receptor activation.

The cellular mechanisms underlying mu opioid facilitation of mossy fiber (MF) long-term potentiation (LTP) and synaptic transmission were investigated in the rat hippocampal slice. Naloxone (10 microM) significantly inhibited the induction of mossy fiber LTP, an effect attributed by Derrick and Martinez [B.E. Derrick, J.L.J. Martinez, Opioid receptor activation is one factor underlying the frequency dependence of mossy fiber LTP induction, J. Neurosci. 14 (1994) 4359-4367] to antagonism of endogenous opioid peptide action. We found that the inhibitory effects of naloxone were not blocked by bicuculline, suggesting that endogenous opioids did not enhance mossy fiber LTP by depressing GABAA inhibition. [d-Ala2, NMePhe4, Glyol5] enkephalin, DAMGO (300 nM), a mu opioid agonist, mimicked the action of endogenous opioids, enhancing both mossy fiber LTP induction and paired-pulse facilitation. DAMGO potentiation of the paired-pulse facilitation of mossy fiber response was also insensitive to bicuculline but was blocked by the mu selective antagonist CTOP. Further analysis of the cellular mechanism showed that the depletion of internal Ca2+ stores by thapsigargin (1 microM), or inhibition of protein kinases by application of staurosporine (1 microM) did not block the DAMGO facilitation of mossy fiber-CA3 synaptic transmission. However, application of phaclofen (100 microM GABAB receptor antagonist or SCH 50911, a more potent GABAB antagonist significantly inhibited the DAMGO effect (49+/-15%; 51+/-19% inhibition, P<0.05). The data indicate that the DAMGO effect on the mossy fiber pathway is partially mediated by a reduction in GABA activation of GABAB receptors. These findings further suggest that endogenous opioid peptides activate mu opioid receptors to facilitate mossy fiber LTP and synaptic transmission in rat hippocampus partially by GABAB receptor-mediated disinhibitory mechanism.

Analgesics, Opioid↗

PubMed1999

Solution structure of potassium channel-inhibiting scorpion toxin Lq2.

Lq2 is a unique scorpion toxin. Acting from the extracellular side, Lq2 blocks the ion conduction pore in not only the voltage- and Ca2+ -activated channels, but also the inward-rectifier K+ channels. This finding argues that the three-dimensional structures of the pores in these K+ channels are similar. However, the amino acid sequences that form the external part of the pore are minimally conserved among the various classes of K+ channels. Because Lq2 can bind to all the three classes of K+ channels, we can use Lq2 as a structural probe to examine how the non-conserved pore-forming sequences are arranged in space to form similar pore structures. In the present study, we determined the three-dimensional structure of Lq2 using nuclear magnetic resonance (NMR) techniques. Lq2 consists of an alpha-helix (residues S10 to L20) and a beta-sheet, connected by an alphabeta3 loop (residues N22 to N24). The beta-sheet has two well-defined anti-parallel strands (residues G26 to M29 and residues K32 to C35), which are connected by a type I' beta-turn centered between residues N30 and K31. The N-terminal segment (residues Z1 to T8) appears to form a quasi-third strand of the beta-sheet.

Amino Acids↗

PubMed1999

Glioblastoma cell-specific expression of fibroblast growth factor receptor-1beta requires an intronic repressor of RNA splicing.

The fibroblast growth factor receptor-1 (FGFR-1) primary transcript is alternatively processed to produce receptors that vary in their ligand affinity and specificity. A high affinity form of this receptor--FGFR-1beta--that lacks the alpha exon is observed on the neoplastic transformation of glial cells. In this study, we have identified a 62-bp sequence located 97 bp downstream from the alpha exon that is required for the exclusion of this exon in a human glioblastoma cell line. Deletion or mutation of this sequence is sufficient to allow enhanced inclusion of the alpha exon or a heterologous exon in glioblastoma cells. Therefore, it would appear that this sequence element plays a key role in the glioblastoma-specific splicing to form FGFR-1beta mRNA.

Base Sequence↗

PubMed1999

Analysis and mapping of gene families encoding beta-1,3-glucanases of soybean.

Oligonucleotide primers designed for conserved sequences from coding regions of beta-1,3-glucanase genes from different species were used to amplify related sequences from soybean [Glycine max (L.) Merr.]. Sequencing and cross-hybridization of amplification products indicated that at least 12 classes of beta-1,3-glucanase genes exist in the soybean. Members of classes mapped to 34 loci on five different linkage groups using an F(2) population of 56 individuals. beta-1,3-Glucanase genes are clustered onto regions of five linkage groups. Data suggest that more closely related genes are clustered together on one linkage group or on duplicated regions of linkage groups. Northern blot analyses performed on total RNA from root, stem, leaf, pod, flower bud, and hypocotyl using DNA probes for the different classes of beta-1,3-glucanase genes revealed that the mRNA levels of all classes were low in young leaves. SGlu2, SGlu4, SGlu7, and SGlu12 mRNA were highly accumulated in young roots and hypocotyls. SGlu7 mRNA also accumulated in pods and flower buds.

Amino Acid Sequence↗

PubMed1999

EAF1 regulates vegetative-phase change and flowering time in Arabidopsis.

We have identified a new locus that regulates vegetative phase change and flowering time in Arabidopsis. An early-flowering mutant, eaf1 (early flowering 1) was isolated and characterized. eaf1 plants flowered earlier than the wild type under either short-day or long-day conditions, and showed a reduction in the juvenile and adult vegetative phases. When grown under short-day conditions, eaf1 plants were slightly pale green and had elongated petioles, phenotypes that are observed in mutants altered in either phytochrome or the gibberellin (GA) response. eaf1 seed showed increased resistance to the GA biosynthesis inhibitor paclobutrazol, suggesting that GA metabolism and/or response had been altered. Comparison of eaf1 to other early-flowering mutants revealed that eaf1 shifts to the adult phase early and flowers early, similarly to the phyB (phytochrome B) and spy (spindly) mutants. eaf1 maps to chromosome 2, but defines a locus distinct from phyB, clf (curly leaf), and elf3 (early-flowering 3). These results demonstrate that eaf1 defines a new locus involved in an autonomous pathway and may affect GA regulation of flowering.

Alleles↗

PubMed1999

[Anatomy of intrahepatic portal branches visualized by three-dimensional imaging analysis of CT arterial portography].

The purpose of this study was to clarify variations in intrahepatic portal branches by means of CT imaging procedures. The subjects were 73 patients, 59 men and 14 women, who ranged in age from 41 to 76 years, with a mean of 63 years. The procedures were as follows. The entire liver was scanned using helical CT during the portal and hepatic venous phases, and 3D images of the portal vein were reconstructed with the volume-rendering technique and the region-growing method. The CT unit was a HITACHI W2000, and the imaging analyzer a Sun Ultra 1. We found that the branching patterns of both the anterior (P5 and P8) and posterior segmental branches (P6 and P7) of the right lobe of the liver could be classified into four types. The caudate branch (P1) and left lateral segmental branches (P2 and P3) were classified into three types, and the interior segmental branch of the left lobe (P4) was classified into two types. The frequency of each pattern was also revealed. These branching types and their frequencies were generally the same as those described in previous reports. Thus, the portal anatomy visualized by these methods indicates that they could be very useful for preoperative examinations or IVR.

Adult↗

PubMed1999

[Cytokine assays for pterygium].

OBJECTIVE: To study the roles of tumor necrosis factor (TNF-alpha) and platelet-derived growth factor (PDGF) in pathogenesis of pterygium. METHODS: Radioimmunoassay and bioactivity assay were used to measure the levels of TNF-alpha and PDGF in tissue culture supernatant of pterygium specimens obtained from 30 patients. RESULTS: The levels of TNF-alpha and PDGF in tissue culture supernatant of pterygium were significantly higher than that of normal conjunctival specimens (t = 2.827, P < 0.01; t = 3.011, P < 0.01), and there was a significant difference between progressive and quiescent cases (t = 2.374, P < 0.05; t = 2.712, P < 0.05). CONCLUSIONS: The abnormal secretion of TNF-alpha and PDGF may be related to the pathogenesis of pterygium.

Adult↗

PubMed1999

[Detecton of circulating antigen in sera from mice infected with Toxoplasma tachyzoites].

AIM: To explore an assay method for the early diagnosis of Toxoplasma infection. METHODS: Serum samples collected from three groups of mice infected experimentally with different doses of Toxoplasma trophozoites were detected for the presence of Toxoplasma circulating antigen by using fast ELISA. RESULTS: Toxoplasma circulating antigen was detected on days 4, 4 and 3 after infection in light, moderate and heavy infection groups, respectively. CONCLUSION: The level of circulating antigen was in parallel with the duration of infection. The determination of circulating antigen is useful in the early diagnosis of Toxoplasma infection.

Animals↗