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C Fang

Publications and source records attributed to C Fang.

At least 37 records · Page 2Linked to original sources
PubMed2000

Chondrocyte-specific enhancer regions in the COMP gene.

The molecular events governing the differentiation of mesenchymal cells into chondrocytes and the expression of cartilage marker genes are poorly understood. Cartilage oligomeric matrix protein is a noncollagenous extracellular matrix protein with a relatively cartilage-specific spatial and temporal expression pattern. To understand the mechanisms controlling chondrocyte-specific expression of cartilage oligomeric matrix protein, we cloned 1.9 kb of the 5' flanking promoter sequence of the murine cartilage oligomeric matrix protein gene and identified two spatially distant cartilage-specific enhancer regions by deletion analysis. One element is situated proximally (proximal positive element: -125 to -75) and a second region is located distally (distal positive region: -1925 to -592) relative to the transcription start site. Interestingly, nucleotides within the proximal positive element are conserved between the mouse and human promoters and resemble consensus sites for the binding of members of the high mobility group class of transcription factors. Defining cartilage-specific regulatory elements in the cartilage oligomeric matrix protein promoter may provide useful molecular probes for identifying transcription factors that control acquisition of the chondrocytic phenotype.

Animals↗
PubMed2000

Molecular cloning, sequencing, and tissue and developmental expression of mouse cartilage oligomeric matrix protein (COMP).

Mouse cartilage oligomeric matrix protein cDNA was cloned and sequenced by a reverse transcription-polymerase chain reaction. The open reading frame encoded a product of 755 amino acids that shares a high degree of identity to and possesses all the characteristic molecular features of both rat and human cartilage oligomeric matrix protein. This suggests that cartilage oligomeric matrix protein is highly conserved during evolution. The clone was 83, 84, and 95% identical to human, bovine, and rat cartilage oligomeric matrix protein cDNA, respectively. In tissues from the adult mouse, cartilage oligomeric matrix protein was expressed not only in cartilage and tendon but in trachea, bone, skeletal muscle, eye, heart, and placenta as well, and no expression was found in other tissues. Immunohistology revealed that cartilage oligomeric matrix was deposited as early as 10 days post coitus in predifferentiated mouse embryo mesenchyme. It was detected in all cartilaginous tissues and in the skeletal muscles of the embryo at day 13. As development progressed, accumulation of cartilage oligomeric matrix protein was marked in the growth plate. At 19 days post coitus, it was prominently deposited in the hypertrophic zone of the growth plate, perichondrium, and periosteum and in the superficial layer of the articular cartilage surface but was absent in the more central areas of the epiphyseal cartilage. The restricted tissue distribution and expression of cartilage oligomeric matrix protein in developing as well as adult mouse tissues suggest the regulation of this protein at the transcriptional level. The findings reported herein are the first detailed characterization of the distribution of cartilage oligomeric matrix protein during early skeletal development of the mouse.

Animals↗
PubMed2000

Expression of cartilage oligomeric matrix protein (COMP) by embryonic and adult osteoblasts.

Cartilage oligomeric matrix protein has been implicated as an important component of endochondral ossification because of its direct effects on chondrocytes. The importance of this protein for skeletal development and growth has been recently illustrated by the identification of mutations in cartilage oligomeric protein genes in two types of inherited chondrodysplasias and osteoarthritic phenotypes: multiple epiphyseal dysplasia and pseudoachondroplasia. In the present study, we report the presence of cartilage oligomeric protein in embryonic and adult osteoblasts. A foot from a 21-week-old human fetus, subchondral bone obtained from knee replacement surgery in an adult patient, and a limb from a 19-day-postcoital mouse embryo were analyzed with immunostaining and in situ hybridization. In the human fetal foot, cartilage oligomeric protein was localized to osteoblasts of the bone collar and at the newly formed bone at the growth plate and bone diaphyses. Immunostaining was performed on the adult subchondral bone and showed positive intracellular staining for cartilage oligomeric protein of the osteoblasts lining the trabecular bone. There was no staining of the osteocytes. Immunostaining of the mouse limb showed the most intense staining for cartilage oligomeric protein in the hypertrophic chondrocytes and in the surrounding osteoblast cells of the developing bone. Cartilage oligomeric protein mRNA and protein were detected in an osteoblast cell line (MG-63), and cartilage oligomeric protein mRNA was detected from human cancellous bone RNA. These results suggest that the altered structure of cartilage oligomeric protein by the mutations seen in pseudoachondroplasia and multiple epiphyseal dysplasia may have direct effects on osteoblasts, contributing to the pathogenesis of these genetic disorders.

Animals↗
PubMed2000

Kupffer cell inactivation alleviates ethanol-induced steatosis and CYP2E1 induction but not inflammatory responses in rat liver.

BACKGROUND/AIMS: Gadolinium chloride inactivates Kupffer cells and alleviates alcohol-induced liver lesions. We investigated the mechanism of gadolinium chloride protection after oral ethanol feeding. METHODS: Rats were maintained ethanol-intoxicated for 6 weeks by feeding ethanol in a low-carbohydrate/high-fat liquid diet. Macrophages were inactivated by intravenous administrations of gadolinium chloride. At termination, liver samples and cell lysates obtained from the periportal and perivenous region were analyzed for histopathology, mRNA expression of endotoxin-associated parameters and cytokines and for enzymes involved in oxidative stress. RESULTS: Ethanol treatment alone caused marked microvesicular/macrovacuolar steatosis and focal inflammation. Gadolinium significantly alleviated pathology, by reducing steatosis but not inflammation. Gadolinium treatment eliminated ED2 immunopositive Kupffer cells, which were larger and more frequent periportally. Ethanol significantly increased the mRNA expression of the endotoxin (LPS) receptor CD14 and the LPS binding protein LBP, but not that of the pro-inflammatory cytokines TNF-alpha and IL-1beta. The mRNA of CD14 was found to be expressed preferentially in the perivenous region, but gadolinium treatment had no significant effect on the expression or the distribution. However, gadolinium significantly moderated the ethanol induction of CYP2E1 and this effect correlated to the degree of steatosis. Ethanol increased glutathione transferase and reduced glutathione peroxidase activity, but these changes persisted after gadolinium treatment. CONCLUSIONS: Our results suggest that gadolinium chloride reduces symptoms of ALD mainly by counteracting steatosis, and that CD14-positive Kupffer cell populations are not involved in gadolinium protection. The strong correlation between pathology and CYP2E1 induction might suggest a steatopathogenic role for this enzyme.

Acute-Phase Proteins↗
PubMed2000

Influence of vegetables on the expression of GSTP1 in humans--a pilot intervention study (Sweden).

BACKGROUND: There are indications that a diet rich in vegetables and/or fruit has a protective effect against several types of diseases, including cancer. Data from experimental and epidemiological studies suggest that antioxidant constituents may provide protection against environmental carcinogens. AIMS: This study investigated the effect of additional vegetables in the diet on the expression of the endogenous antioxidant enzyme GSTP1 in human lymphocytes. METHODS: Six subjects were given an addition of mixed vegetables to their normal diet for a period of three weeks. The expression of GSTP1 protein and mRNA in lymphocytes was measured by Western blot and RT competitive PCR. RESULTS: After the intervention all six subjects had lower levels of GSTP1 mRNA, and five of the six subjects had lower GSTP1 protein levels. This suggests that increased vegetable intake decreases GSTP1 expression, possibly through the supply of additional antioxidants.

Adult↗
PubMed2000

Inhibition of ethanol-induced liver disease in the intragastric feeding rat model by chlormethiazole.

The purpose of this investigation was to assess the effect of chlormethiazole treatment on liver damage in the experimental rat intragastric ethanol-feeding model of alcoholic liver disease. Chlormethiazole has been used in the treatment of alcoholic withdrawal and has been shown to inhibit cytochrome P4502E1. Since treatment of experimental alcoholic liver disease with CYP2E1 inhibitors had an ameliorating effect on liver injury in the rat, chlormethiazole was used to see if it had a similar effect. Rats fed ethanol for 2 months had significantly less liver injury when chlormethiazole was added to the diet, fed intragastrically. The CYP2E1 apoprotein levels, which were increased by ethanol feeding, were also increased when chlormethiazole was fed with ethanol. Chlormethiazole inhibited the increase in the ethanol-induced CYP2E1 activity in vivo, as measured by chlorzoxazone 6-hydroxylation, but did not affect the level of CYP2E1 apoprotein. Likewise, the reduction in proteasome proteolytic enzyme activity produced by ethanol feeding was blunted in chlormethiazole-fed rats. These results support the conclusion that chlormethiazole treatment partially protects the liver from injury by inhibiting CYP2E1 activity in vivo.

Animals↗
PubMed2000

[The changes in serum antibody level after immunization with HFRS vaccine].

OBJECTIVES: To observe the changes in serum antibody level after mass immunization with vaccine against hemorrhagic fever with renal syndrome (HFRS) and to evaluate its efficacy and effectiveness in the prevalent areas. METHODS: Healthy people aged 16 to 60 years in the villages were recruited as study subjects, excluding those suffered from HFRS previously, going out for more than nine months and those with contraindications, and were randomly allocated into immunization and control groups with 10,460 and 16,159 persons, respectively. Specific IgG antibody was determined with indirect immunofluorescent assay (IFA) and neutralizing antibody (NA) was determined with micro CPE method. RESULTS: Two weeks after the full-course immunization, sero-conversion rate for IFA reached 100% in those sero-negative before immunization, with a 95% confidence interval of 96.3 - 100.0%, and that for NA 44.4%, with a 95% CI of 22.0% - 69.0%. Geometric mean titer (GMT) were 72.1 and 4.6 for IFA and NA, respectively. Booster immunization was provided for them one year later. Positivity of IFA and NA was 28.6%, 83.3%, 75.0%, 53.1%, 22.6% and 14.8%, 55.6%, 35.0%, 31.3%, 26.0%, before booster immunization, two weeks, one year, one and a half years, two years after booster immunization, respectively. CONCLUSION: HFRS vaccine had good immunogenicity, but its duration of serum antibody sustenance was relatively short.

Adolescent↗
PubMed2000

[Quantitative and immunohistochemical analysis of endothelin-1 in oral submucous fibrosis].

OBJECTIVE: This experiment is carried out to investigate roles of endothelin-1 (ET-1), a potent vasoconstrictor and mitogenic peptide, in fibrosis and collagen production. METHODS: 30 cases of oral submucous fibrosis(OSF), 10 cases of oral lichen planus (OLP) and 10 cases of normal control, were studied by means of SABC immunohistochemical method with polyclonal antisera to ET-1 and using image pattern analysis technique. RESULTS: 1. Oral mucosa from OLP and normal control expressed very little ET-1. By contrast, there was striking expression of ET-1 in oral mucosa from OSF; 2. The cellular content of ET-1 in OSF was significantly higher than that in normal control (P < 0.01), while the cellular content of ET-1 in the early and moderate stages OSF were significantly higher than that in the advanced stage(P < 0.01); 3. The cellular content of ET-1 in the early and moderate stages OSF were significantly higher than that in OLP(P < 0.01); 4. There was a significantly positive correlation between cellular contents of epithelia and interstitial(P < 0.05). CONCLUSION: These findings suggest that cell-specific expression of ET-1 may play a role in pathogenesis of OSF.

Endothelin-1↗
PubMed1999

Role of APC in the selection of immunodominant T cell epitopes.

Following antigenic challenge, MHC-restricted T cell responses are directed against a few dominant antigenic epitopes. Here, evidence is provided demonstrating the importance of APC in modulating the hierarchy of MHC class II-restricted T cell responses. Biochemical analysis of class II:peptide complexes in B cells revealed the presentation of a hierarchy of peptides derived from the Ig self Ag. Functional studies of kappa peptide:class II complexes from these cells indicated that nearly 20-fold more of an immunodominant epitope derived from kappa L chains was bound to class II DR4 compared with a subdominant epitope from this same Ag. In vivo, T cell responses were preferentially directed against the dominant kappa epitope as shown using Ig-primed DR4 transgenic mice. The bias in kappa epitope presentation was not linked to differences in class II:kappa peptide-binding affinity or epitope editing by HLA-DM. Rather, changes in native Ag structure were found to disrupt presentation of the immunodominant but not the subdominant kappa epitope; Ag refolding restored kappa epitope presentation. Thus, Ag tertiary conformation along with processing reactions within APC contribute to the selective presentation of a hierarchy of epitopes by MHC class II molecules.

Amino Acid Sequence↗
PubMed1999

Effect of low frequency ultrasound on the in vitro percutaneous absorption of clobetasol 17-propionate.

The effect of low frequency ultrasound (20 kHz) on the permeation of clobetasol 17-propionate (CP) through skin (sonophoresis) was studied. The ultrasound was applied at either continuous or discontinuous modes and at different intensities. The results showed that low frequency ultrasound significantly enhanced the permeability of CP across hairless mouse skin in vitro. Delivering the same amount of ultrasonic energy in different modes of application markedly influenced the flux and skin residual of CP. The on/off discontinuous ultrasound had greater enhancement on CP permeation than the continuous ultrasound. The results of skin histopathology and permeation experiment using various membranes demonstrate that both disordering of stratum corneum and convective flow resulted from the cavitation effect were responsible for sonophoretic enhancement of CP. The permeation of CP through hair follicles and sweat ducts was susceptible to the application of ultrasound.

Administration, Topical↗
PubMed1999

Effect of chronic coadministration of endotoxin and ethanol on rat liver pathology and proinflammatory and anti-inflammatory cytokines.

To better understand how gut-derived endotoxins influence alcohol-induced liver injury and the expression of inflammatory cytokines a new animal model was developed. After 2 weeks on a modified ethanol-containing liquid diet, some rats also were infused with endotoxin via osmotic minipumps for 4 additional weeks. Ethanol diet alone increased plasma endotoxin threefold to 9.3 pg/mL. Endotoxin infusion increased the levels to 388 and 513 pg/mL in controls and ethanol-fed animals, respectively. Panlobular macrovesicular and microvesicular steatosis and inflammatory foci were observed in livers from both ethanol- and ethanol-endotoxin-treated animals, but there was no significant potentiation by endotoxin. Only minor changes, mainly polymorphonuclear infiltration, were seen in animals treated with endotoxin alone although the messenger RNA (mRNA) expression of both proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta) and anti-inflammatory cytokines IL-4 and IL-10 were markedly increased, as shown by competitive polymerase chain reaction (PCR) analysis using cyclophilin as standard. The effect of endotoxin infusion on cytokine mRNA expression in ethanol-fed animals was not significantly different. Expression of transforming growth factor beta1 (TGF-beta1) mRNA was increased twofold by ethanol, eightfold by endotoxin, but only threefold by ethanol-endotoxin treatment. The mRNA expression of lipopolysaccharide binding protein (LBP) and CD14 endotoxin receptor was not significantly increased by chronic endotoxin treatment, contrasting with the marked elevation observed after acute endotoxin challenge. These results suggest that the tolerance observed despite sustained hepatic expression of proinflammatory cytokines is counteracted by the anti-inflammatory cytokines and by down-regulation of CD14 and LBP. Furthermore, a similar adaptation may occur in alcoholics with continuous endotoxemia.

Acute-Phase Proteins↗
PubMed1999

Localization and expression of cartilage oligomeric matrix protein by human rheumatoid and osteoarthritic synovium and cartilage.

Synovium and cartilage from patients with osteoarthritis or rheumatoid arthritis were analyzed for expression of cartilage oligomeric matrix protein. Immunostaining of synovium with antiserum to cartilage oligomeric matrix protein demonstrated positive staining in both diseases. In osteoarthritis, there was positive staining within the synovial cells and immediately subjacent connective tissue, with less intense staining in the deeper connective tissue. In rheumatoid arthritis, there was less intense staining within the synovial cells and marked intense staining in the deeper connective tissue. In situ hybridization performed with an antisense digoxigenin-labeled riboprobe to human cartilage oligomeric matrix protein confirmed the presence of cartilage oligomeric matrix protein mRNA in the cells of the synovial lining in both types of synovium. Quantitative polymerase chain reaction with a cartilage oligomeric matrix protein MIMIC demonstrated increased cartilage oligomeric matrix protein mRNA in rheumatoid cartilage and synovium as compared with osteoarthritic cartilage and synovium, respectively; mRNA levels in rheumatoid synovium were similar to those from osteoarthritic chondrocytes. As a result of the high expression of cartilage oligomeric matrix protein from rheumatoid synovium, inflammatory synovium should be considered as a potential tissue source of cartilage oligomeric matrix protein in any investigation of biological markers of cartilage metabolism. The upregulated expression of cartilage oligomeric matrix protein in inflammatory tissues suggests its in vivo regulation by cytokines.

Arthritis, Rheumatoid↗
PubMed1999

Circumferential resection and "Z"-shape plastic end-to-end anastomosis of canine trachea.

OBJECTIVE: To prevent anastomotic stricture of the trachea. METHODS: Forty young mongrel dogs, weighing 5-7 kg, were randomly divided into two groups: experimental group and control group, with 20 dogs in each group. Four tracheal rings were removed from each dog. In the experimental group, two "Z"-shape tracheoplastic anastomoses were performed on each dog, one on the anterior wall and the other on the membranous part of the trachea. In the control group, each dog received only simple end-to-end anastomosis. Vicryl 3-0 absorbable suture and OB fibrin glue were used for both groups. All dogs were killed when their body weight doubled. RESULTS: The average sagittal stenotic ratio were 1.20 +/- 0.12 for the experimental group and 0.83 +/- 0.05 for the control group. The average cross-sectional area stenotic ratio were 0.90 +/- 0.12 and 0.69 +/- 0.09 and T values were 8.71 and 4.57 for the two groups (P < 0.01). CONCLUSION: "Z"-shape tracheoplasty is superior to simple end-to-end anastomosis in preventing anastomotic stricture of canine trachea.

Anastomosis, Surgical↗
PubMed1999

[Look back on the tenth revision of the international classification of diseases (ICD - 10)].

Following the 6th revision, the 10th revision of ICD is another revision of significance as a milestone. The major change is the replacement of pure numerical form of code structure by alphabet - numerical one. There is also progress in the classification thinking in ICD-10, marking the progress toward maturity of the thinking in disease classification. Descriptions are given to the general condition, revision of its contents, the conception of classified 'family' of disease classification, and the new renewal mechanism in the 10th revision.

Classification↗
PubMed1998

Role of the transmembrane sequence of spleen focus-forming virus gp55 in erythroleukemogenesis.

The membrane glycoprotein encoded by the env gene of either the polycythemia- or anemia-inducing spleen focus-forming virus (SFFVp or SFFVa, respectively) is responsible for the induction of erythroleukemia in mice. It has been shown that the SFFVp glycoprotein, gp55, interacts with the erythropoietin receptor (EPO-R) and promotes EPO-independent proliferation of an EPO-R-expressing hematopoietic cell line, Ba/F3 (Li et al., Nature 343:762, 1990). We show here that when residues within the transmembrane (TM) sequence of an SFFVp gp55 are altered based on the sequences of the anemia-inducing gp55s by a methionine-to-isoleucine (M-I) substitution, a di-leucine deletion (dLL), or both, the resulting mutants display an attenuated phenotype that resembles an SFFVa: they induce milder erythroproliferative disease without polycythemia in vivo and are unable to promote EPO-independent cell proliferation in vitro. The dLL mutation directly interferes with EPO-R binding by decreasing the affinity of gp55 for the receptor. On the other hand, the M-I mutation hampers the full mitogenic activation of EPO-R while having no effect on receptor binding and asserts a dominant negative effect over the wild-type SFFVp gp55. Two other sequence changes within the TM sequence did not affect the biological activities of the SFFVp gp55. These results indicate that the TM sequence of the SFFV env glycoprotein plays a prominent role in SFFV-induced erythroleukemogenesis through its influence on the mitogenic activation of EPO-R.

3T3 Cells↗
PubMed1998

Alzheimer's beta-amyloid vasoactivity: identification of a novel beta-amyloid conformational intermediate.

The beta-amyloid (A beta) peptide has previously been shown to enhance phenylephrine or endothelin-1 induced constriction of aortic rings in vitro. The characteristics of A beta vasoactivity (dose, fragment length, timing) suggest that the mechanism is distinct from A beta cytotoxicity. To identify which properties of A beta determine its biological activity on vessels, we investigated a number of A beta analogues and fragments, individually and in combination, including those that are known to be associated with Alzheimer's disease (A beta(1-42)) and hereditary cerebral hemorrhage with amyloidosis--Dutch type (A beta(22Q)(1-40)). The vasoactivity appears to be related to the conformation adopted by the peptide in solution. The beta-pleated sheet rich A beta(1-42) and A beta(22Q)(1-40) were each less vasoactive than the mainly random coil wild type A beta(1-40). However, the most vasoactive A beta peptides were combinations which contain mixtures of random coil and beta-sheet structure. The finding that peptides containing low or high levels of beta-pleated conformation are less vasoactive than those containing intermediate amounts of this structural motif allows us to propose the existence of a transitional form between random coil and beta-pleated that is the vasoactive species of A beta. This is the first time that A beta conformational intermediates have been identified and a biological activity associated with them.

Amyloid beta-Peptides↗