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Biomedical subjects

W Lin

Publications and source records attributed to W Lin.

At least 91 records · Page 5Linked to original sources

Transient and permanent resolution of ischemic lesions on diffusion-weighted imaging after brief periods of focal ischemia in rats : correlation with histopathology.

BACKGROUND AND PURPOSE: The early ischemic lesions demonstrated by diffusion-weighted imaging (DWI) are potentially reversible. The purposes of this study were to determine whether resolution of initial DWI lesions is transient or permanent after different brief periods of focal brain ischemia and to evaluate histological outcomes. METHODS: Sixteen rats were subjected to 10 minutes (n=7) or 30 minutes (n=7) of temporary middle cerebral artery occlusion or sham operation (n=2). DWI, perfusion-weighted imaging (PWI), and T(2)-weighted imaging (T(2)WI) were performed during occlusion; immediately after reperfusion; and at 0.5, 1.0, 1.5, 12, 24, 48, and 72 hours after reperfusion. After the last MRI study, the brains were fixed, sectioned, stained with hematoxylin and eosin, and evaluated for neuronal necrosis. RESULTS: No MRI or histological abnormalities were observed in the sham-operated rats. In both the 10-minute and 30-minute groups, the perfusion deficits and DWI hyperintensities that occurred during occlusion disappeared shortly after reperfusion. The DWI, PWI, and T(2)WI results remained normal thereafter in the 10-minute group, whereas secondary DWI hyperintensity and T(2)WI abnormalities developed at the 12-hour observation point in the 30-minute group. Histological examinations demonstrated neuronal necrosis in both groups, but the number of necrotic neurons was significantly higher in the 30-minute group (95+/-4%) than in the 10-minute group (17+/-10%, P<0.0001). CONCLUSIONS: Transient or permanent resolution of initial DWI lesions depends on the duration of ischemia. Transient resolution of DWI lesions is associated with widespread neuronal necrosis; moreover, permanent resolution of DWI lesions does not necessarily indicate complete salvage of brain tissue from ischemic injury.

Animals↗

Effects of fluid management on edema volume and midline shift in a rat model of ischemic stroke.

BACKGROUND AND PURPOSE: The purpose of this study was to investigate the effects of fluid management on brain water content (BW) and midline shift (MLS) after a focal cerebral ischemic insult. METHODS: A suture model was used to induce focal cerebral ischemia for 90 minutes (n=44). The rats were randomly assigned to 3 groups 2. 5 hours after reperfusion: dehydration (n=24), control (n=8), or hydration (n=12). BW was obtained with the wet-dry weight method 24 hours after middle cerebral artery (MCA) occlusion. In addition, MRI were obtained (n=31) 24 hours after the onset of ischemia so that the ratio of hemispheric volumes ipsilateral (IH) and contralateral (CH) to the infarct and the extent of MLS could be obtained. RESULTS: Across the range from moderate dehydration to intravascular volume expansion with isotonic saline, BW of the IH increased linearly as a function of change in body weight (r(2)=0.89), whereas few changes in relation to body weight were observed in CH, indicating a preferential effect of fluid management on the infarcted hemisphere. Furthermore, the hemispheric volume ratio (IH/CH) and MLS also increased in relation to changes in body weight. However, paradoxical increases in BW, IH/CH, and extent of MLS were observed in comparison with controls when severe dehydration was produced with high-dose mannitol. CONCLUSIONS: Changes in ischemic BW by fluid management correlated closely with changes in body weight except when high-dose mannitol was used. Mannitol, as a dehydrating agent, may be associated with bimodal effects, with a high dose aggravating ischemic BW.

Animals↗

Immunogenicity and safety of an inactivated hepatitis A vaccine in Taiwanese adults and children.

The safety and immunogenicity of an inactivated hepatitis A vaccine (AVAXIM, 160 antigen units) was evaluated in 190 subjects: 50 children aged from 2 to 5 years, 70 children aged from 6 to 17 years and 70 adults aged from 18 to 30 years in a monocentric, open, non-controlled, phase III trial conducted in Taipei, Taiwan from December 1996 to October 1997. The vaccine was administered intramuscularly, with a two-dose schedule 6 months apart. Clinical adverse events were monitored during the seven days following each injection. Hepatitis A virus (HAV) antibody titers were measured by modified radioimmunoassay on the day of inclusion and four weeks after both the first dose and booster injection. Among the 190 subjects who received the first dose, 174 (91.6%) were initially HAV seronegative and 16 (8.4%) were HAV seropositive at inclusion. One hundred and seventy-four subjects (91.6%) received the booster dose and completed the study. One month after the first dose, all the subjects, whatever the age, presented HAV antibody titers over 20 mIU/ml. In children (2 to 17 years), the GMT was 136 mIU/ml at week 4 and 7,906 mIU/ml four weeks after the booster dose. In adults (> or = 18 years), GMT values were 93 mIU/ml at week 4 and 3,655 mIU/ml four weeks after the booster. These results show a strong anamnestic response to the second dose of vaccine and are compatible with long-term antibody persistence in each age group. The vaccine was safe and well tolerated. No vaccine-related serious adverse event occurred. No immediate reaction occurred. The majority of the reactions were reported by adults after the primary injection. Local reactions (pain and redness) were reported by 9.0% and 4.0% of the subjects after the primary and the booster doses, respectively. Systemic reactions (mainly myalgia/arthralgia or asthenia) affected less than 10% of the subjects after the first dose and less than 3% after the booster. Results from this study in a Taiwanese population are consistent with those obtained with the same vaccine in previous European studies in children and adults, and suggest that AVAXIM (160 AU) is suitable for use in all subjects aged over 2 years.

Adolescent↗

[Summary of 42 patients with acute aortic occlusion].

OBJECTIVE: To summarize the experience of diagnosis and treatment in 42 patients with acute aortic occlusion. METHODS: Clinic data were analyzed. The emergency surgery of retrograde catheter thrombolectomy of transaortic thromboembolism were done, and the propathy and complications were treated simultaneously. RESULTS: Normal blood flow were restored just after operation. Four patients died of myonephropathic metabolic syndrome and heart failure. 38 patients were followed up for 0.5 to 4 years. Among them, 36 were followed for more than 1 year. A better effectiveness was achieved in 28 patients. Two patients died of recurrence accompanied with heart failure. CONCLUSION: Early diagnosis and treatment may increase the curative ratio and decrease the mortality in patients with acute aortic occlusion.

Acute Disease↗

[Separation of thoracopagus conjoined twins].

OBJECTIVE: To sam up the experience in diagnosis and separation of a thoracopagus conjoined twins. METHODS: Thoracopagus twin boys were admitted for evaluating possible separation and repair at age of 21 days. X-rays, CT, MRI, and ultrasound scan of torso showed separate and normal gastrointestinal and biliary tracts, little shared vasculature, but shared pericardium, diaphragm and liver parenchyma. The chest junctions were at the sternums and the most inferior two pairs of the ribs. One twin (twin A) had a protruding heart with VSD and ASD. RESULTS: The infants were separated successfully at age of 45 days. The chest defect of twin A was closed with aid of a polyester pericardial patch graft and a silicon supporter. Managing anesthesia met some difficulty in understanding of the anatomical and physiological consequences of the complex anomaly. They are had brain edema and multiple system organ failures after separation, and were cured under intensive care. They are alive and well 6 months after surgery. CONCLUSIONS: Careful preoperative evaluation proper anesthesia and surgical technique, and postoperative care are essential to the successful separation of the conjoined twins.

Anesthesia↗

[The effect of TGF-beta1 on the expression of plasminogen activator inhibitors 1 and matrix metalloproteinases in cultured rat mesangial cells transfected with antisense-TGF-beta1 vector].

OBJECTIVE: To study the plasminogen activator inhibitors 1 (PAI-1) and matrix metalloproteinases (MMPs) expression on the cultured rat mesangial cells (MsC) transfected with antisense-TGF-beta1 vector. METHODS: Lipofectin method was used to transfect antisense-TGF-beta1 vector into MsC, Western blot and Northern blot analysis for detecting TGF-beta1 peptides and antisense TGF-beta1 RNA level. The expression of PAI-1 and MMPs were determined by Northern blot, Western bolt and zymography assay. RESULTS: The MsC clone (MAT1) which blocked the expression of TGF-beta1 peptides was successfully established. The MsC clone showed increased expression of MMP-2 mRNA protein and enzyme activity of MMP-2, 3, 7. However, expression of PAI-1 mRNA of MsC clone was suppressed. CONCLUSION: Blocked expression of TGF-beta1 can upregulate the expression of MMP-2, 3, 7 and downregulate the expression of PAI-1 mRNA in MsC. It is possible that antisense-TGF-beta1 may be used for gene therapy in glomerular diseases.

Animals↗

[Evidence for TCR Vbeta clonal expansion T cells in a patient with cGVHD after unrelated donor BMT].

OBJECTIVE: To investigate the distribution and clonal expansion of TCR Vbeta subfamily T cells in patients with cGVHD after unrelated donor bone marrow transplantation (BMT). METHODS: The CDR3 of 24 TCR Vbeta subfamily genes were amplified from peripheral mononuclear cells of a patient with cGVHD after unrelated BMT. RT-PCR was used for detection of the distribution of TCR Vbeta repertoire, and the PCR products were further analyzed by genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR VbetaT cells. RESULTS: Only 8 Vbeta subfamily T cells could be identified in the patient, clonal expansion T cells could be found of Vbeta2, 3, 8 and 13 subfamilies. CONCLUSIONS: The skew distribution and clonal expansion of TCR Vbeta subfamily T cells could be found in the patient. It may relate to the initiation of cGVHD.

Acute Disease↗

[Clinical study on treatment of atherosclerosis obliterans by integrated traditional Chinese and Western medicine].

OBJECTIVE: To investigate the mechanism in treating atherosclerosis obliterans (ASO) with integrated traditional Chinese and western medicine (TCM-WM). METHODS: The changes in level of vasoactive factors, such as endothelin (ET), nitric oxide (NO), apoprotein and the iconographic changes in angiography before and after treatment were observed in 40 patients of ASO. RESULTS: All patients had high levels of plasma ET and low level of NO, abnormal metabolism apoproteins, and showed the signs of trunk artery obstruction and marked decrease of collateral circulation in affected limb. After treatment, the plasma ET, NO and apoprotein levels recovered to normal range in 30 markedly effective cases, as compared with those before treatment, the difference was significant (P < 0.01) and their collateral circulation in the affected limb significantly increased, showing obvious improvement of peripheral circulation. CONCLUSION: The therapeutic mechanism of TCM-WM in treating ASO might be related with its effects on vascular endothelium, lipid metabolism adjustment, collateral circulation establishment and microcirculation improvement.

Aged↗

[Construction of cDNA library of Mortierella and screening of delta 9 fatty acid desaturase cDNA sequence].

Mortierella species have potential for fermentative production of polyunsaturated fatty acids including gamma-linolenic acid, Arachidonic acid and EPA, etc. In order to clone genes encoding enzymes in the unsaturated fatty acid biosynthetic pathway, cDNA library of Mortierella was constructed using lambda gt 10 vector. Using cDNA encoding conserved region of delta 9 fatty acid desaturase gene as probe, Mortierella cDNA library was screened. After two rounds of screening one positive clone was identified which has insert length of larger than 1.6 kb.

Base Sequence↗

Cysteinyl-tRNA formation: the last puzzle of aminoacyl-tRNA synthesis.

With the exception of the methanogenic archaea Methanococcus jannaschii and Methanobacterium thermoautotrophicum deltaH, all organisms surveyed contain orthologs of Escherichia coli cysteinyl-tRNA synthetase (CysRS). The characterization of CysRS-encoding (cysS) genes and the demonstration of their ability to complement an E. coli cysSts mutant reveal that Methanococcus maripaludis and Methanosarcina barkeri, two other methanogenic archaea, possess canonical CysRS proteins. A molecular phylogeny inferred from 40 CysRS sequences indicates that the CysRS of M. maripaludis and Methanosarcina spp. are specific relatives of the CysRS of Pyrococcus spp. and Chlamydia, respectively. This result suggests that the CysRS gene was acquired by lateral gene transfer in at least one euryarchaeotic lineage.

Amino Acid Sequence↗

The human REV1 gene codes for a DNA template-dependent dCMP transferase.

DNA is frequently damaged by various physical and chemical agents. DNA damage can lead to mutations during replication. In the yeast Saccharomyces cerevisiae, the damage-induced mutagenesis pathway requires the Rev1 protein. We have isolated a human cDNA homologous to the yeast REV1 gene. The human REV1 cDNA consists of 4255 bp and codes for a protein of 1251 amino acid residues with a calculated molecular weight of 138 248 Da. The human REV1 gene is localized between 2q11.1 and 2q11.2. We show that the human REV1 protein is a dCMP transferase that specifically inserts a dCMP residue opposite a DNA template G. In addition, the human REV1 transferase is able to efficiently and specifically insert a dCMP opposite a DNA template apurinic/apyrimidinic (AP) site or a uracil residue. These results suggest that the REV1 transferase may play a critical role during mutagenic translesion DNA synthesis bypassing a template AP site in human cells. Consistent with its role as a fundamental mutagenic protein, the REV1 gene is ubiquitously expressed in various human tissues.

Amino Acid Sequence↗

Functional characterization of the CFTR R domain using CFTR/MDR1 hybrid and deletion constructs.

To improve our insight into the structure and function of the CFTR R domain, deletion and hybrid constructs in which different parts of the R domain were deleted or replaced by the MDR1 linker domain, and vice versa, were made. Replacement of the linker domain by the R domain did not result in a decrease and replacement of the CFTR R domain by the linker domain did not result in an increase of maturation efficiency, when compared to the respective wild-type proteins. This indicates that the R domain is not responsible for the high degree of degradation observed for CFTR translation products in the ER, but rather the overall structure or sequences located outside the R domain. Replacing the C-terminal part of the R domain (amino acids 780-830) by the MDR1 linker domain resulted in the appearance of PKA-dependent whole cell chloride currents which were not significantly different from wild-type CFTR currents. This might indicate that the PKA sites present in the linker domain are functional and that not the exact sequence of the C-terminal part of the R domain is important, but rather the presence of PKA sites and the length. Moreover, when this hybrid construct was PKC-stimulated, chloride currents were activated. Although these PKC-induced currents were lower than the PKA-induced ones, this again indicates that the linker domain is functional in this hybrid construct. Taken together, these results suggest that the MDR1 linker domain can substitute for part of the regulatory domain of the CFTR protein.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Preparation and in vitro characterization of HSA-mPEG nanoparticles.

Surface modified human serum albumin (HSA) nanoparticles with a size of approximately 150 nm in diameter were prepared from a PEG-HSA conjugate, methoxy-polyethylene glycol modified human serum albumin (HSA-mPEG) using a coacervation method and crosslinked with glutaraldehyde. The zeta-potential of the surface modified nanoparticles was significantly lower than that of unmodified HSA nanoparticles. The existence of a hydrated steric barrier surrounding the nanoparticles was confirmed by electrolyte and pH induced flocculation tests. The surface modified nanoparticles showed a reduced plasma protein adsorption on the particle surface compared with unmodified particles.

Animals↗

Identification and gene structure of a novel human PLZF-related transcription factor gene, TZFP.

A novel cDNA clone was identified through yeast two-hybrid experiments. Following cross-examination between the cDNA clones, EST clones, and the cosmid clone, we could digitally assemble a new zinc finger transcription factor gene. This predicted gene has a cDNA size of about 1960 bp and is translated into a 487-amino-acid protein. According to database analysis, this gene contains three C2H2 zinc finger motifs and is highly related to human PLZF (promyelocytic leukemia zinc finger protein). The full-length coding region of the gene was isolated, and its sequences were confirmed by DNA sequencing. Interestingly, one splicing variant lacking exon III was also identified. Northern blot analysis revealed that this gene is mainly expressed in human testis. In conclusion, we have identified a new member of the PLZF zinc finger protein family, the testis zinc finger protein (TZFP), which is mainly expressed in testis tissue.

Amino Acid Sequence↗

Fast repair of hydroxy radical purine deoxynucleotide adducts by phenylpropanoid glycosides and their derivatives from Chinese herbs.

DNA damaged by oxygen radicals has been implicated as a causative event in a number of degenerative diseases, including cancer and aging. So it is very significant to look for ways in which either oxygen radicals are scavenged prior to DNA damage or damaged DNA is repaired to supplement the cells' inadequate repair capacity. The repair activities and reaction mechanism of phenylpropanoid glycosides (PPGs) and their derivatives, isolated from Chinese folk medicinal herbs, towards both dGMP-OH* adducts and dAMP-OH* adducts were studied with the pulse radiolytic technique. On pulse irradiation of nitrous oxide saturated 2 mM dGMP or dAMP aqueous solution containing one of the PPGs or their derivatives, the transient absorption spectra of the hydroxyl adduct of dGMP or dAMP decayed with the formation of that of phenoxyl radicals of PPGs or their derivatives within several decades of microseconds after electron pulse irradiation. The result indicated that dGMP or dAMP hydroxyl adducts can be repaired by PPGs or their derivatives. The rate constants of the repair reactions were deduced to be 0.641-1.28 x 10(9) M(-1) s(-1) for dGMP-OH* and 0.2-0.491 x 10(9) M(-1) s(-1) for dAMP-OH*, which positively correlated to the number of phenolic hydroxyl groups in the glycoside structure. A deeper understanding of this new repair mechanism may help researchers to design strategies to prevent and/or intervene more effectively in free radical related diseases.

DNA Adducts↗

Fast repair of deoxynucleotide radical cations by phenylpropanoid glycosides (PPGs) and their analogs.

The repair effects on deoxynucleotide radical cations of phenylpropanoid glycosides (PPGs) and their analogs, isolated from a Chinese folk medicinal herb, were studied using the pulse radiolysis technique. The radical cations of deoxynucleotides were formed by the reaction of SO4*- with deoxynucleotides. On pulse irradiation of a nitrogen saturated deoxynucleotide aqueous solution containing 20 mM K2S2O8, 200 mM t-BuOH and one of the PPGs or their analogs, the transient absorption spectra of the radical cations of nucleotide decayed with the formation of those of the radical cation of PPGs or their analogs within several tens of microseconds after electron pulse irradiation. The result indicates that deoxynucleotide radical cations can be repaired by PPGs or their analogs. The rate constants of the repair reactions were determined to be 0.48-1.1 x 10(9), 0.64-1.80 x 10(9) and 2.12-4.4 x 10(9) M(-1) s(-1) for dAMP, dGMP and dCMP radical cations respectively. It is obvious that the rate constants of the repair reaction depend on the number of phenolic hydroxyl groups contained in the PPGs and their analogs. A deeper understanding of this new repair mechanism will undoubtedly help researchers design strategies to prevent and/or intervene more effective in free radical related diseases.

Cations↗

Identification of a virulence-associated protein homolog gene and ISRa1 in a plasmid of Riemerella anatipestifer.

Riemerella anatipestifer is the causative agent of polyserositis of ducks and geese. We have previously reported that a 3.9-kb plasmid, pCFC1, carries protein genes (vapD1 and vapD2) that are similar to virulence-associated genes of other bacteria. In the present study, we report the complete sequence of a second plasmid of 5.6 kb, pCFC2. pCFC2 has a 28% G-C content and three large open reading frames (ORFs). One of the ORFs (designated asVapD1) encodes a polypeptide that shares 53.9, 53.9, 48.3, 48.3 and 46.1% identity with virulence-associated proteins of Dichelobacter nodosus, Actinobacillus actinomycetemcomitans, Neisseria gonorrhoeae, Helicobacter pylori and Haemophilus influenzae, respectively. The second ORF encodes a putative DNA replication protein (RepA3) with 309 amino acids and a molecular mass of approximately 36 kDa. A novel insertion sequence (IS) element, designated ISRa1, was found on the plasmid pCFC2. ISRa1 was flanked by 15-bp imperfect inverted repeats (only one mismatched nucleotide). ISRa1 contained an ORF encoding a putative transposase of 292 amino acids. Southern blot analysis indicated that in R. anatipestifer strains examined, ISRa1 was present with 2-20 copies (at least). ISRa1 displayed a sequence approximately 35% homologous to the putative IS982 and RSBst-alpha from Lactococcus lactis ssp. cremoris SK11 and Bacillus stearothermophilus CU21. Three hybridization patterns of genomic DNA of eight R. anatipestifer strains with an ISRa1 probe indicated that ISRa1 might be a useful tool for epidemiological studies.

Amino Acid Sequence↗

Nuclear magnetic resonance and oxygen affinity study of cesium binding in human erythrocytes.

We investigated the interaction of the cesium ion (Cs(+)) with the anionic intracellular components of human red blood cells (RBCs); the components studied included 2,3-bisphosphoglycerate (BPG), ADP, ATP, inorganic phosphate (P(i)), carbonmonoxy hemoglobin (COHb), and RBC membranes. We used spin-lattice (T(1)) and spin-spin (T(2)) (133)Cs NMR relaxation measurements to probe Cs(+) binding, and we found that Cs(+) bound more strongly to binding sites in BPG and in RBC membranes than in any other intracellular component in RBCs at physiologic concentrations. By using James-Noggle plots, we obtained Cs(+) binding constants per binding site in BPG (66 +/- 8 M(-1)), ADP (19 +/- 1 M(-1)), ATP (25 +/- 3 M(-1)), and RBC membranes (55 +/- 2 M(-1)) from the observed T(1) values. We also studied the effect of Cs(+) on the oxygen (O(2)) affinity of purified Hb and of Hb in intact RBCs in the absence and in the presence of BPG. In the absence of BPG, the O(2) affinity of Hb decreased upon addition of Cs(+). However, in the presence of BPG, the O(2) affinity of Hb increased upon addition of Cs(+). The O(2) affinity of Cs(+)-loaded human RBCs was larger than that of Cs(+)-free cells at the same BPG level. (31)P NMR studies on the pH dependence of the interaction between BPG and Hb indicated that the presence of Cs(+) resulted in a smaller fraction of BPG available to bind to the cleft of deoxyHb. Our NMR and O(2) affinity data indicate that a strong binding site for Cs(+) in human RBCs is BPG. A partial mechanism for Cs(+) toxicity might arise from competition between Cs(+) and deoxyHb for BPG, thereby increasing oxygenation of Hb in RBCs, and thus decreasing the ability of RBCs to give up oxygen in tissues. The presence of Cs(+) at 12.5 mM in intact human RBCs containing BPG at normal concentrations did not, however, alter significantly the O(2) affinity of Hb, thus ruling out the possibility of Cs(+)-BPG interactions accounting for Cs(+) toxicity in this cell type.

2,3-Diphosphoglycerate↗