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Biomedical subjects

J Lin

Publications and source records attributed to J Lin.

At least 343 records · Page 19Linked to original sources

[Recognition of transient evoked otoacoustic emissions].

OBJECTIVE: To measure the cochlear function more accurately, seeking for a novel method to recognize transient evoked otoacoustic emission (TEOAE) from the recorded signal. METHODS: Time-frequency domain method was used to effectively recognize TEOAE and stimulating artifact by their different short time Fourier transformation (STFT). RESULTS: The STFT of stimulating artifact features decayed rapidly in the air by exponential rule. The frequency distribution of TEOAE was similar to that of the stimulating signal, but the latency and shifting of TEOAE showed a complicated patern. The STFTs of stimulating artifact and TEOAE varied significantly. CONCLUSION: STFT recognition method can separate TEOAE and stimulating artifact effectively so as to improve the accuracy of TEOAE recognition.

Adolescent↗

[Effects of cyclosporin A combined with T4 on collagen IV production, coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression in cultured human mesangial cells].

OBJECTIVE: To investigate the effects of single cyclosporin A (CsA), tripcholorlide (T4) or both on renal fibrosis. METHODS: Collagen IV concentrations in the supernatants of human mesangial cells (HMCs) in culture were measured by ELISA. Collagen type IV alpha 1 mRNA (Coll alpha 1 IV mRNA) and transforming growth factor beta 1 (TGF beta 1) mRNA expression were tested by RT-PCR. RESULTS: CsA regulated HMC collagen IV production biphysically, i.e., 1.0 microgram/ml CsA increased collagen synthesis while 0.01-0.1 microgram/ml CsA decreased its formation. T4 enhanced collagen IV production in a dose dependent manner. Combination of the two drugs in lower doses could reach the same degree of immunosuppression as a higher dose of CsA did but with less fibrogenesis. Both CsA and T4 up-regulated HMC Coll alpha 1 (IV) mRNA and TGF beta 1 mRNA expression. The degree of Coll alpha 1 (IV) mRNA expression in different groups were in accordance with that of TGF beta 1 mRNA. CONCLUSIONS: Both CsA and T4 regulate extracellular matrix formation which may be mediated by TGF beta 1. Combination of the two drugs in lower dose results in less fibrosis than a higher dose of CsA.

Cells, Cultured↗

[Treatment of late thoracolumbar fracture by transpedicular decompression using "PUMC trephine" 26 cases report].

OBJECTIVE: To introduce a new operation to treat late thoracolumbar fracture cases with neurological deficit or with kyphosis. METHODS: 26 cases were treated with "PUMC trephine" (designed by professor Ye Qibin). RESULTS: The kyphosis was (29.7 +/- 14.5) degrees Cobb's angle in pre-operation and (10 +/- 11.8) degrees in post-operation (P < 0.01). The angle of correction was (19.7 +/- +9.9) degrees. The height of the anterior edge of traumatic-vertebra was (16.3 +/- 5.7) mm in pre-operation and (22.2 +/- 6.3) mm in post-operation. The corrected height was (6.5 +/- 3.2) mm. The neurological deficit also got a better recovery in 17 of 19 cases, 12 of 15 cases obtained complete cure of incontinence. 11 of 15 case had sexual function recovery after following up 1-9 years (average 5.2 years) cases. CONCLUSIONS: Using "PUMC trephine" procedure was a new, simple, safety and less-traumatic technique for treatment of late thoracolumbar fracture through posterior approach.

Adult↗

An experimental study on homoharringtonine liposome and glaucoma filtration surgery.

PURPOSE: To observe the inhibitory action of homoharringtonine liposome during the healing process of wounds in the filtering sites. METHODS: Posterior sclerectomies were performed in 14 rabbits. Postoperatively one eye of each rabbit received subconjunctival injections of HH liposome and fellow eye received saline injection in a randomized masked fashion. RESULTS: Fourteen days after operation the IOP of experimental eyes reduced significantly (P < 0.01) as compared with the controlled eyes, and the number of remaining filtering blebs increased noticeably (P < 0.05). Pathohistological examination revealed that the number of fibroblasts per square micron in the filtering sites and the thickness of the scars in the center of the filtering sites of the experimental eyes were less than those of the controlled eyes. No serious ocular toxic and side effects were found. CONCLUSION: This experiment suggest that homoharringtonine liposome can markedly inhibit the scar formation of filtering sites after glaucoma filtering surgery, promote the formation of filtering blebs, and provide an experimental evidence for its clinical use in future.

Animals↗

[Expanded polytetrafluoroethylene with different pore diameter for keratoprosthesis cell ingrowth and corneal metabolism].

PURPOSE: (1). To study the relationship between the different pore diameter of the expanded polytetrafluoroethylene, (ePTFE) and the rate of cellular ingrowth in cornea; (2). To evaluate the influence of implant on corneal metabolism. METHODS: Two kinds of ePTFE disk with 45 microns and 28 microns pore diameter were implanted after a free-hand intralamellar dissection. The eyes were examined by slit-lamp biomicroscopy up to 7 months postoperatively. The corneas were removed and examined by light and transmission electron microscopy. RESULTS: Stromal fibroblasts penetrate, proliferate, and synthesize collagen in the pore of these two kinds of ePTFE, while the rate of cellular ingrowth and collagen deposition was greater in the ePTFE with 45 microns pore than 28 microns pore. Lipid deposits occurred in 3 eyes with 45 microns pore and in 5 eyes with 28 microns pore (P > 0.05). Dicks remained in corneas for 7 months without any extrusion in all eyes. CONCLUSION: Stromal fibroblasts can penetrate, proliferate, and synthesize collagen in the pore of both two kind of ePTFE, while cells penetrate greatly in the larger pore. Dicks interfere corneal metabolism in some degree. The ePTFE with 45 microns pore diameter, 200 mm thick, 84% porosity is a suitable material for the skirt of the keratoprosthesis.

Animals↗

Fresh amniotic membrane transplantation for conjunctival surface reconstruction.

PURPOSE: To determine whether fresh human amniotic membrane can be used to reconstruct the conjunctival detect created during symblepharon lysis. METHODS: Forty-two eyes of 39 consecutive patients with eye burns and Stevens-Johnson syndrome were randomized to accept fresh or preserved human amniotic membrane transplantation (AMT) during the period of severe scarring. Impression cytology was performed in 12 eyes with normal tear secretion which received fresh AMT. RESULTS: During a mean follow-up of 11 months (range, 6 to 18 months), thirty-five patients (37 eyes) showed successful ocular surface reconstruction and resolution of motility restriction while four patients (2 eyes with fresh AMT, 3 eye with preserved AMT) with minimal recurrence of symblepharon. There was no significant difference statistically between two groups (Chi-square test). Amniotic epithelial cells can survive about three months after being transplanted onto ocular surfaces with normal tear secretion. CONCLUSION: Both fresh and preserved human amniotic membrane can be considered an ideal alternative substrate for conjunctival surface reconstruction during removal of severe symblepharon.

Adolescent↗

Rapid diagnosis of chlamydial conjunctivitis in laboratory.

PURPOSE: To evaluate the techniques of rapid and accurate diagnosis of chlamydial conjunctivitis. METHODS: Total 100 conjunctivitis patients (200 eyes) were studied. Twenty-two of 100 cases were diagnosed as chlamydial conjunctivitis. The infected epithelia were scraped from tarsal conjunctiva of both eyes and stained separately with Giemsa (100 cases) and immunofluorescence (anti-chlamydial antigen monoclonal antibody, 100 cases). RESULT: In immunofluorescent staining, 38 cases were seen positive staining and 62 were negative. In Giemsa staining, 29 were positive, and 71 were negative. In 22 cases with clinical diagnosis of chlamydial conjunctivitis, 13 cases were confirmed, and 9 were excluded by immunofluorescent staining. Technically, immunofluorescent and Giemsa stain takes 45 and 40 minutes, respectively. CONCLUSION: Comparing to Giemsa stain, 38 of 100 scraping specimens were positive (38%) by immunofluorescent staining, 29 of 100 per cent were positive by Giemsa staining. Giemsa staining takes 5 minutes less than immunofluorescent staining (40 versus 45 minutes), however, the positive staining in immunofluorescent staining is much easier to be recognized than Giemsa staining.

Adolescent↗

[The experimental investigation of epithelial healing in rabbit central corneal alkali wounds].

PURPOSE: To observe the morphologic changes of epithelium in the alkali wounds at the central corneal of rabbits. METHODS: 38 rabbits weighing 2.5 kg were used. A alkali wound of the corneas was performed in the right eye of 38 New Zealand white rabbits with round filter paper, 8 mm in diameter, which were soaked in 0.5 mol/L NaOH for 5 seconds and then were placed centrally on the cornea for 60 seconds. The corneal burns were examined and photographed after the staining of 2% fluoresent staining with a slit lamp every day. The excised corneas were examined in the light microscope, electron microscope. RESULTS: The epithelial cells had rapidly disappeared in the burned area. The epithelial cells regrowth started at 8 hours following the burn, and 1-2 layers epithelial cells covered the burned area by 24 hours. The epithelial defects occured again on the 4th day. The desmosome and microvilli of the regenerating epithelium had a reduced number. There was no basement membrane between the regenerating epithelium and the strom. There were increased intercellular spaces between the regenerating epithalial cells. CONCLUSION: The epithelial healing were achieved through the division and migration of epithelial cells at the margin of the wound in rabbit central corneal alkali wound. The decrease of the junctions and the increase of intercellular spaces between the regenerating epithalial cells may be responsible for the second epithelial defects.

Alkalies↗

Failure and complication following surgical treatment of scoliosis--an analysis of 101 cases.

Complications occur frequently after surgical treatment of scoliosis. In order to prevent from them effectively, 101 cases with failure and postoperative complications were analysed. They included rod fracture in 22 cases (15 Harrington rods, 4 Zielke rods, and 3 Luque rods); recurrence of curve severity in 12 cases; broken or loossened luque wires in 15 cases; loss of thoracic kyphosis (flat back) in 6 cases; progressive kyphosis with or without paraplegia following incorrect posterior decompression in 5 cases; and increased unbalance of shoulders after instrumentation in 2 cases due to neglect of the tilting of the first thoracic vertebra. Infection occurred in 8 cases (incision infection 7 cases, deep wound infection in 1 case); and pneumothorax in 1 case. They were induced by biomechanical factors in 23 cases (22.8%), incorrect selections of indications in 29 cases (28.7%), operational mistakes in 37 cases (36.6%), internal fixation factors in 15 cases (14.9%). The authors hold that there are quite a lot of factors leading to occurrence of complications and the effective way for prevention from them is to understand the factors and main technical points related to internal fixation.

Equipment Failure↗

Fractionation factors and activation energies for exchange of the low barrier hydrogen bonding proton in peptidyl trifluoromethyl ketone complexes of chymotrypsin.

NMR investigations have been carried out of complexes between bovine chymotrypsin Aalpha and a series of four peptidyl trifluoromethyl ketones, listed here in order of increasing affinity for chymotrypsin: N-Acetyl-L-Phe-CF3, N-Acetyl-Gly-L-Phe-CF3, N-Acetyl-L-Val-L-Phe-CF3, and N-Acetyl-L-Leu-L-Phe-CF3. The D/H fractionation factors (phi) for the hydrogen in the H-bond between His 57 and Asp 102 (His 57-Hdelta1) in these four complexes at 5 degreesC were in the range phi = 0.32-0.43, expected for a low-barrier hydrogen bond. For this series of complexes, measurements also were made of the chemical shifts of His 57-Hepsilon1 (delta2,2-dimethylsilapentane-5-sulfonic acid 8.97-9. 18), the exchange rate of the His 57-Hdelta1 proton with bulk water protons (284-12.4 s-1), and the activation enthalpies for this hydrogen exchange (14.7-19.4 kcal.mol-1). It was found that the previously noted correlations between the inhibition constants (Ki 170-1.2 microM) and the chemical shifts of His 57-Hdelta1 (delta2, 2-dimethylsilapentane-5-sulfonic acid 18.61-18.95) for this series of peptidyl trifluoromethyl ketones with chymotrypsin [Lin, J., Cassidy, C. S. & Frey, P. A. (1998) Biochemistry 37, 11940-11948] could be extended to include the fractionation factors, hydrogen exchange rates, and hydrogen exchange activation enthalpies. The results support the proposal of low barrier hydrogen bond-facilitated general base catalysis in the addition of Ser 195 to the peptidyl carbonyl group of substrates in the mechanism of chymotrypsin-catalyzed peptide hydrolysis. Trends in the enthalpies for hydrogen exchange and the fractionation factors are consistent with a strong, double-minimum or single-well potential hydrogen bond in the strongest complexes. The lifetimes of His 57-Hdelta1, which is solvent shielded in these complexes, track the strength of the hydrogen bond. Because these lifetimes are orders of magnitude shorter than those of the complexes themselves, the enzyme must have a pathway for hydrogen exchange at this site that is independent of dissociation of the complexes.

Animals↗

Inhibition of dipeptidyl peptidase IV by fluoroolefin-containing N-peptidyl-O-hydroxylamine peptidomimetics.

Dipeptidyl peptidase IV (EC 3.4.14.5; DPP IV), also known as the leukocyte differentiation antigen CD26 when found as an extracellular membrane-bound proline specific serine protease, cleaves a dipeptide from the N terminus of a polypeptide chain containing a proline residue in the penultimate position. Here we report that known (Z)-Ala-psi[CF=C]-Pro dipeptide isosteres 1 and 2, which contain O-acylhydroxylamines, were isolated as diastereomeric pairs u-1, l-1, and l-2. The effect of each diastereomeric pair as an inhibitor of human placental dipeptidyl peptidase DPP IV has been examined. The inhibition of DPP IV by these compounds is rapid and efficient. The diastereomeric pair u-1 exhibits very potent inhibitory activity with a Ki of 188 nM. Fluoroolefin containing N-peptidyl-O-hydroxylamine peptidomimetics, by virtue of their inhibitory potency and stability, are superior to N-peptidyl-O-hydroxylamine inhibitors derived from an Ala-Pro dipeptide.

Crystallography, X-Ray↗

De novo design, synthesis, and biological activities of high-affinity and selective non-peptide agonists of the delta-opioid receptor.

On the basis of the structure-activity relationships of delta-opioid-selective peptide ligands and on a model of the proposed bioactive conformation for a potent and selective, conformationally constrained delta-opioid peptide ligand [(2S, 3R)-TMT1]DPDPE, a series of small organic peptide mimetic compounds targeted for the delta-opioid receptor have been designed, synthesized, and evaluated in radiolabeled ligand binding assays and in vitro bioassays. The new non-peptide ligands use piperazine as a template to present the most important pharmacophore groups, including phenol and phenyl groups and a hydrophobic moiety. This hydrophobic group was designed to mimic the hydrophobic character of the D-Pen residues in DPDPE, which has been found to be extremely important for increasing the binding affinity and selectivity of these non-peptide ligands for the delta-opioid receptor over the mu-opioid receptor. Compound 6f (SL-3111) showed 8 nM binding affinity and over 2000-fold selectivity for the delta-opioid receptor over the mu-opioid receptor. Both enantiomers of SL-3111 were separated, and the (-)-isomer was shown to be the compound with the highest affinity for the delta-opioid receptor found in our study (IC50 = 4.1 nM), with a selectivity very similar to that observed for the racemic compound. The phenol hydroxyl group of SL-3111 turned out to be essential to maintain high affinity for the delta-opioid receptor, which also was observed in the case of the delta-opioid-selective peptide ligand DPDPE. Binding studies of SL-3111 and [p-ClPhe4]DPDPE on the cloned wild-type and mutated human delta-opioid receptors suggested that the new non-peptide ligand has a binding profile similar to that of DPDPE but different from that of (+)-4-[((alphaR)-alpha(2S,5R)-4-allyl-2, 5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide (SNC-80), another delta-opioid-selective non-peptide ligand.

Animals↗

Identification of four genes in endothelial cells whose expression is affected by tumor cells and host immune status--a study in ex vivo-isolated endothelial cells.

A spontaneously metastasizing, well-defined mouse lymphoma was chosen as an in vivo model to study the effect of tumor-host interaction on gene expression in liver sinusoidal endothelial cells. Forty-nine bovine aortic endothelial cell (BAEC) genes, recently isolated by a differential screening approach of a cDNA library enriched for tumor necrosis factor-alpha (TNF-alpha) suppressed genes, were investigated. Four of these genes were finally selected because they were affected differentially by host immuno-competence, TNF-alpha, and tumor cells. Sequence analysis showed them to encode the bovine polyubiquitin (A4), elongation factor 1alpha (B2), the acidic ribosomal phosphoprotein PO (C3), and the ribosomal protein S2 (E10). Gene expression was analyzed by dot-blot or Northern blot analysis. TNF-alpha and tumor cell conditioned supernatant suppressed the genes additive in BAEC but not in other endothelial cells except for bovine capillary endothelial cells. Ex vivo-isolated liver endothelial cells of tumor-bearing syngeneic DBA/2 mice showed strong downregulation of these four genes in comparison to normal control values. In contrast, endothelial cells of tumor-bearing immuno-incompetent Balb/c (nu/nu) mice showed no downregulation but upregulation of these genes. Consistently, all four genes were also downregulated when BAEC were incubated with supernatants derived from ex vivo-isolated liver metastases from immuno-competent but not from -incompetent mice. Thus, the expression of a group of genes involved in protein translation and processing was more profoundly altered in endothelial cells in vivo than in vitro, suggesting that microenviromental factors and cell-cell and cell-matrix interactions play an important role.

Amino Acid Sequence↗

The relationship of the -5, -8, and -24 variant alleles in African Americans to triosephosphate isomerase (TPI) enzyme activity and to TPI deficiency.

In 424 African-American and 75 white subjects, we found that the -5 (TPI 592 A-->G), -8 (TPI 589 G-->A), and -24 (TPI 573 T-->G) variants in the triosephosphate isomerase (TPI) gene occurred frequently (41.0%) in the African-American subjects but did not occur in the whites. These data suggest that this set of polymorphisms may turn out to be one of the higher-incidence molecular markers of African lineage, a surprising finding because others had reported that these nucleotide substitutions were restricted to a small subset of African Americans who had been characterized as TPI-deficiency heterozygotes. Additionally, we investigated the relationship of these variants to TPI-enzyme activity. Although the variant substitutions (occurring in three haplotypes: -5 alone, -5 -8, and -5 -8 -24) were associated with moderate reduction in enzyme activity, severe-deficiency heterozygotes could not be identified with certainty, and none of the haplotypes were restricted to subjects with marked reduction of enzyme activity. Three subjects were homozygous for the -5 -8 haplotype, a finding inconsistent with the putative role of this haplotype as the cause of a null variant incompatible with life in homozygotes. Despite these findings, the possibility remains that the -5 -8 or -5 -8 -24 haplotypes may in some instances contribute to compound heterozygosity and clinical TPI deficiency.

Alleles↗

Correlations of the basicity of His 57 with transition state analogue binding, substrate reactivity, and the strength of the low-barrier hydrogen bond in chymotrypsin.

The basicity of His 57-Nepsilon2 within the low-barrier hydrogen-bonded (LBHB) diad His 57-Asp 102 and the 1H NMR chemical shift of the LBHB proton in tetrahedral, hemiketal complexes of chymotrypsin with peptidyl trifluoromethyl ketones (peptidyl-TFKs) have been studied. The following results were obtained with various peptidyl-TFKs at 5 degrees C: N-Ac-Gly-DL-Phe-CF3, pKa = 11.1 and deltaLBHB = 18.7 ppm; N-Ac-L-Val-DL-Phe-CF3, pKa = 11.8 and deltaLBHB = 18.9 ppm; N-Ac-L-Leu-DL-Val-CF3, pKa = 10.3 and deltaLBHB = 18.9 ppm; and N-Ac-L-Leu-DL-naphthyl-CF3, pKa = 10.9 and deltaLBHB = 19.0 ppm. Results for peptidyl-TFKs with Phe in the P1 position and N-Ac, N-Ac-Gly, N-Ac-L-Val, and N-Ac-L-Leu in the P2 position were well correlated with literature values for inhibition constants Ki and kcat/Km for the corresponding peptidyl methyl esters. The plot of log Ki versus the apparent pKa of His 57-Nepsilon2 displayed a slope of -0.77, and that of log kcat/Km for peptidyl methyl esters versus the pKa of His 57-Nepsilon2 in corresponding peptidyl-TFK complexes gave a slope of 0.68. The slope of a plot of pKa versus deltaLBHB was 3.7, and that of log kcat/Km for peptidyl methyl ester substrates versus deltaLBHB for the corresponding peptidyl-TFK-chymotrypsin complexes was 2.7. A plot of log Ki versus deltaLBHB displayed a slope of -3.0. These plots indicated that the pKa of His 57 and substrate reactivity were correlated with increasing strength of the low-barrier hydrogen bond. The apparent pKa of His 57-Nepsilon2 for the chymotrypsin-N-Ac-L-Leu-DL-Phe-CF3 complex is 10.6 at 25 degrees C, whereas it is 12.0 at 5 degrees C [Cassidy, C. S., Lin, J. L., and Frey, P. A. (1997) Biochemistry 36, 4576-4584]. The apparent discrepancy is likely to be due to a temperature dependence in the cooperative ionization of His 57 in peptidyl-TFK complexes, which appears to be coupled to inhibitor dissociation, hydration and ionization of free peptidyl-TFK, ionization of Ile 16, and a conformational change.

Animals↗

Characterization and functional analysis of the porcine lactoferrin gene promoter.

Lactoferrin, a ferric binding glycoprotein found in milk, can possibly prevent microbial infection of the mammary gland and gastrointestinal tract. To define the regulation of the porcine lactoferrin gene (pLTF), we cloned its 5'-flanking region from a porcine liver genomic library and analyzed the 5' upstream region of approx. 4kb, two exons, and an intron. The transcription start site was localized by primer extension to residue G, which is 41 nucleotides upstream from the ATG start codon. The pLTF 5'-flanking region possesses several putative cis-acting regulatory elements found in both housekeeping and inducible genes; to define their function, they were inserted into a chloramphenicol acetyltransferase reporter construct. The region up to -156 sufficed for basic promoter activity, whereas the region up to -780 was required for maximal promoter activity in porcine testis cells (STcells), kidney cells (PK15 cells) and human mammary epithelial cells (HBL-100 cells). Detailed analysis of this proximal region by DNase I footprinting and electrophoretic mobility shift assays reveals that the ubiquitous factors SP1, AP2 and the mammary gland-specific factor (MGF) might play significant roles in regulating the transcription of the pLTF gene.

Animals↗