Biomedical subjects

C W Chi

Publications and source records attributed to C W Chi.

At least 73 records · Page 4Linked to original sources

PubMed1997

Purification and sequence determination of a new neutral mammalian neurotoxin from the scorpion Buthus martensii Karsch.

A new neutral mammalian neurotoxin, designated BmK M4, with an isoelectric point (pI) of 7.6 and a relatively low toxicity (LD50 = 4.0 +/- 0.25 microgram/g mice, i.v.) was purified from the venom of scorpion Buthus martensii Karsch (BmK). The complete amino acid sequence of the toxin composed of 64 amino acid residues was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidomethylated protein (up to 30 amino acid residues) and its peptide fragments degraded by lysylendopeptidase or Staphylococcus aureus Va protease. The calculated mol. wt based on the amino acid composition was 7001. By comparison with the sequences of other basic BmK mammalian neurotoxins, it was concluded that the weaker toxicity and lower pI value of BmK M4 might be the result of mutations H10E, R18G and K28D. The sequence comparison of BmK M4 with an acidic toxin, BmK M8, showed that the weakest toxicity and acidic property of BmK M8 may be the consequence of mutations K8D, D53A, V55E and V59E. The substitution of 21 Gly in BmK M4 for Tyrin other BmK toxins may also be of importance. In their tertiary structures, these mutated charged residues are mainly distributed in the surface (face B) that is roughly opposite to the "conserved hydrophobic surface" (face A) proposed by Fontecilla-Camps et al. in 1982. Therefore the toxin-receptor interaction may take a multiposition mode.

Amino Acid Sequence↗

PubMed1997

The CDNA and genomic DNA sequences of a mammalian neurotoxin from the scorpion Buthus martensii Karsch.

The cDNA library of venomous glands of the scorpion Buthus martensii Karsch (BmK) was constructed. A cDNA encoding a mammalian neurotoxin corresponding to the known alpha-type toxin, BmK M1, was amplified by polymerase chain reaction (PCR) and cloned, and its full-length sequence was determined. The open reading frame encoded the precursor of BmK M1 with 84 amino acid residues, including a signal peptide of 19 residues, a mature toxin of 64 residues and an additional C-terminal residue Arg which might be cleaved off by proteinase postprocessing immediately after protein synthesis. Based on the determined cDNA sequence and using the total DNA of the scorpion as a template, the gene of BmK M1 was also amplified by PCR and sequenced. The genomic DNA sequence revealed an intron of 408 base pairs present within the signal peptide region. Both the intron and exon of BmK M1 share about 75% similarity with those of AaH I' another alpha-type mammalian neurotoxin in the scorpion Androctonus australis Hector.

Amino Acid Sequence↗

PubMed1997

Clarification of an uncertain intron within the cDNA sequences of arrowhead proteinase inhibitors A and B.

An uncertain intron of 87 bp within the cDNA sequences of arrowhead proteinase inhibitors A and B was clarified. By site-directed mutation with either a stop codon inside the uncertain intron or mutated codons at both its 5' and 3' ends, it was proved that there was neither a translation intron nor a protein intron present in the cDNA sequences of proteinase inhibitors A and B. The primary structure of inhibitor B was then reexamined by mass spectrometry molecular weight determination and partial amino acid sequencing. A 38 residue peptide was derived by degradation of inhibitor B with lysylendopeptidase, and purified, which was not found in the previous work, and its N-terminal part was none other than the missed 29 residue peptide encoded by the uncertain intron. The 38 residue peptide was very hydrophobic, while the 29 residue peptide it included was even more hydrophobic. The N-terminal part of the missed peptide was also aligned within a BrCN-degraded fragment of the inhibitor. In this paper the cause of the overlooking of this 29 residue peptide in the previous work and some unexpected problems which arose during the former sequence analysis are explained.

Amino Acid Sequence↗

PubMed1997

Phase II study of megestrol acetate in the treatment of hepatocellular carcinoma.

This is a report of a phase II study of megestrol acetate (160 mg/day, orally) in the treatment of hepatocellular carcinoma (HCC). Forty-six patients with advanced HCC were studied and tumour response, changes in appetite, bodyweight, a feeling of well-being, survival and toxicity were evaluated. Thirty-two patients were able to be evaluated for response; there were no complete responders or partial responders. Twelve patients (38%) had stable disease and seven of these patients had a minor response with a median size reduction in the tumour of 18%. Twenty patients (62%) had progressive disease. Five of 24 (21%) patients had a median reduction in alpha-fetoprotein levels of 59 ng/mL. The overall median survival was 4 months (range 1 week to 27 months). Twenty of 32 (62%) patients had an increased appetite and feeling of well-being. Fourteen of 22 (64%) patients had a median lean bodyweight gain of 5 kg (range 1-14 kg). Toxicities were minimal. Tests for glucocorticoid receptors were performed in 10 patients. Four of five patients who were positive for glucocorticoid receptors in the tumour had a stable disease and all five patients who were negative for glucocorticoid receptors had progressive disease. Megestrol acetate had no significant effect on the tumour in HCC patients. However, megestrol acetate is useful in the palliative management of HCC patients, with improvements in appetite, bodyweight and a feeling of well-being with minimal side effects. Some patients had stable disease, a minor reduction of tumour size and a prolonged survival after megestrol acetate treatment and this response may be related to the presence of glucocorticoid receptors in the HCC tumour.

Aged↗

PubMed1997

Risk factors for Alzheimer's disease: a case-control study.

The extent of current knowledge of risk factors for Alzheimer's disease (AD) is still limited. We conducted a case-control study of 98 AD patients and 98 age- and sex-matched controls. Light smoking had a decreased risk for AD (AD: 2.0%, controls: 21.4%, odds ratio = 0.10, p = 0.003), whereas daily smoking showed a trend to increase the risk for AD (AD: 45.9%, controls: 26.5%, odds ratio = 1.73, p = 0.08). After multivariable analyses factors associated with AD included the presence of apolipoprotein epsilon4 allele, and the duration of well water consumption.

Aged↗

PubMed1997

Using monoclonal antibodies to characterize a sequential epitope on the group I allergen of Bermuda grass pollen.

BACKGROUND: Cyn d 1, the group I allergen of Bermuda grass pollen, had been purified and characterized. METHODS: A sequential B cell epitope on Cyn d 1 was studied with monoclonal antibodies (MoAbs). Cyn d 1 was cleaved by Achromobacter protease I into fragments, and the resulting peptides were fractionated on reversed-phase columns before being reacted with anti-Cyn d 1 MoAbs in a radioimmunoassay. A Cyn d 1 fragment recognized by its MoAb was selected for Edman degradation. A synthetic peptide was constructed according to the determined sequence. RESULTS: The epitope on Cyn d 1 recognized by MoAb 18-53 was found to be conformation independent, since its activity was not changed after sodium periodate, guanidine or urea treatment. The enzyme-cleaved fragment containing this epitope was determined to be DVDKPPFDGMTACGNEPIF which corresponds to the N-terminal 46-64 residues of Cyn d 1. The presence of this sequence in the epitope recognized by MoAb 18-53 was demonstrated by enzyme immunoassay and further confirmed by inhibition of binding enzyme immunoassay with synthetic peptides. Some cross-reactivity with the N-terminal 45-63 residues of Lol p 1 was also found. CONCLUSIONS: The primary structure of a sequential epitope on Cyn d 1 was determined, and its activity was confirmed with peptides synthesized according to the determined sequence.

Allergens↗

PubMed1997

[Effect of intrathecal or intracerebroventricular administrition of OFQ on pain threshold and acpuncture analgesia in rats].

Orphanin FQ (OFQ) is a newly discovered 17-amino-acid peptide capable of inducing hyperalgesia. In the present study, the effects of OFQ on basal pain threshold and acupuncture anlgesia (AA) in rats were observed using the tail-flick test. It was found that intrathecal (i.t.) or intracerebroventricular (i.c.v.) administrition of 0.1 microgram OFQ had no effect on basal pain threshold of rats, while 1 microgram OFQ could lower the threshold. However, OFQ at both the doses (0.1 or 1.0 microgram) administered by either i.t. or i.c.v. injection could antagonize AA with that occuring in the brain being more prominent then in the spinal cord. When the rats were repeatedly treated with antisense oligonucleotide to block synthesis of OFQ receptor, pain threshold increased significantly. At such instance, when the OFQ was combined with acupuncture, the effect of AA showed no obvious change. The above results show that the OFQ at small dose has no effect on pain threshold but can lower it at larger dose; while in both cases OFQ can antagonize AA.

Acupuncture Analgesia↗

PubMed1996

Modification of mitomycin C-induced clastogenicity by Terminalia catappa L. in vitro and in vivo.

The water extract of the leaves of Terminalia catappa L. was tested for inhibition of mitomycin C-induced micronuclei in CHO-K1 cells. The simultaneous and pre-treatment of CHO-K1 cells with T. catappa extract (75 and 150 micrograms/ml) significantly (P < 0.05) suppressed mitomycin C-induced micronuclei. Furthermore, gastric intubation of T. catappa extract (4.8 and 24 mg/animal per day) to male ICR mice for 8 days significantly (P < 0.01) reduced mitomycin C-induced micronuclei in peripheral blood. In addition, T. catappa dose dependently inhibited lipid peroxidation in vitro and TPA-induced hydrogen peroxide formation in human mononuclear leukocytes. The anticlastogenic effects of T. catappa in vitro and in vivo may be attributed to its antioxidative potential.

Animals↗

PubMed1996

Paclitaxel-induced apoptosis in human gastric carcinoma cell lines.

BACKGROUND: Gastric cancer is one of the most common cancers in Asia. Chemotherapy and radiation therapy have had limited success. Recently, paclitaxel has been found to be effective against a variety of cancers, including lung, breast, ovary, melanoma, and prostate. Whether paclitaxel is effective in the treatment of gastric cancer is not known and is worthy of investigation. METHODS: Human gastric carcinoma cell lines NUGC-3 and SC-M1 were examined for response to paclitaxel treatment. Cancer cells were treated with paclitaxel (0.001, 0.01, 0.1, and 1 microM) for 1-3 days. Cell number was counted by hemocytometer and cell viability was determined by the trypan blue exclusion method. Cell cycle progression and expression of proliferating cell nuclear antigen (PCNA) were examined by flow cytometry. The percentage of apoptotic cells was determined after staining with hematoxylin and eosin. RESULTS: Paclitaxel was cytotoxic to the two human gastric carcinoma cell lines examined. The growth-inhibiting dose was 0.01 microM. Paclitaxel-treated gastric carcinoma cells were arrested mainly in G2/M phases before apoptosis. However, treatment with 0.01 microM of paclitaxel resulted in a decrease of cells at G0/G1 phases without an increase of cells at G2/M phase indicating that paclitaxel was also cytotoxic to gastric carcinoma cells at G0/G1 phases. In addition, the expression of PCNA was significantly increased in 0.1 and 1 microM paclitaxel-treated cells, suggesting that DNA repair was increased in these cells. CONCLUSIONS: Paclitaxel is effective in growth inhibition of gastric carcinoma cell lines in clinically attainable concentrations. Our results suggest that paclitaxel is a potential chemotherapeutic drug for gastric carcinoma.

Antineoplastic Agents, Phytogenic↗

PubMed1996

Immunohistochemical study of arginase in cancer of the stomach.

High levels of arginase have been detected in gastric adenocarcinoma. To examine the hypothesis that this is due to macrophage infiltration into the tumour, we localized the cellular distribution of arginase by immunohistochemical staining. We examined gastric adenocarcinomas and their corresponding normal tissues (n = 45), leiomyomas (n = 2), leiomyosarcomas (n = 3), human gastric adenocarcinoma cell lines (n = 3), and benign gastric ulcers (n = 4) by the avidin-biotin-peroxidase complex technique. Macrophages with strong arginase immunoreactivity were observed infiltrating both gastric normal and cancer tissues. No arginase immunoreactivity was observed in normal mucosal gland, muscular and serosal tissues or benign gastric ulcers. The immunoreactivity of arginase was positive but heterogeneous in most specimens of gastric adenocarcinoma (62.2%) and was absent from gastric intestinal metaplasia, leiomyomas and leiomyosarcomas. Among the 28 neoplasms with arginase immunoreactivity, scattered immunoreactivity was also noted in adjacent dysplastic glands in 12 (42.8%) specimens. Arginase immunoreactivity was observed in all three gastric cancer cell lines. Arginase is present in the cytoplasm but not in the nucleus. These data suggest that the high arginase levels in adenocarcinoma cancer tissues originate largely from cancer cells.

Adenocarcinoma↗

PubMed1996

Tissue potassium, selenium, and iron levels associated with gastric cancer progression.

The contents of 10 minor and trace elements in histologically confirmed gastric adenocarcinomas and their corresponding normal gastric mucosal tissues obtained from 39 patients at the time of gastric resection were simultaneously determined by instrumental neutron activation analysis. Specimens were irradiated by reactor neutrons and subsequently subject to direct analysis using a high-resolution HPGe gamma-spectrometer. Univariate analysis revealed that gastric cancer tissues had significantly higher concentrations of Fe, K, Mg, Na, Rb, Se, and Zn than normal gastric mucosal tissues. However, multivariate analysis found that Fe, K, and Se were independent elements that associated with gastric cancer. Upon further evaluation of their clinical significance, we found a high tissue K level was related to lymphatic duct metastasis. High Se tissue levels were linked to intestinal type adenocarcinoma. A positive correlation was found between high Fe levels and vascular involvement. These findings suggest that Fe and K are associated with gastric cancer progression. Se is involved in carcinogenesis of stomach in high-risk areas. The mechanisms that underlie the corresponding pathohistological features deserve further study.

Adenocarcinoma↗

PubMed1996

Oxidative damage to DNA induced by areca nut extract.

In Taiwan, people chew betel quid which contains tender areca nut with husk. In other countries, people prefer ripe and dried areca nut without husk. In this study, we compared the reactive oxygen species-induced oxidative DNA damage in isolated DNA and CHO-K1 cells between treatments with tender areca nut extract (ANE) and ripe ANE. Incubation of these two ANE preparations with isolated DNA generated 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in an alkaline environment in a dose-dependent manner. Ripe ANE generated higher levels of 8-OH-dG compared to tender ANE. The addition of iron(II) (100 microM) resulted in 1.4- and 3.1-fold increases of 8-OH-dG when incubated with 1 mg/ml each of tender and ripe ANE. In testing the effect of ANE to cellular DNA, CHO-K1 cells were used for its documented sensitivity to reactive oxygen species. In CHO-K1 cells, ripe ANE was more cytotoxic than tender ANE following an 18-h incubation. The cytotoxicity to CHO-K1 cells was positively correlated with the formation of 8-OH-dG following tender (r=0.97) and ripe (r=0.91) ANE treatment. Addition of the iron chelating agent o-phenanthroline (10 and 20 microM) to cells prior to ri ANE exposure significantly increased (p<0.05) the survival of CHO-K1 cells. In addition, ripe ANE induced dichlorofluorescein-mediated fluorescence which indicated the formation of hydrogen peroxide in CHO-K1 cells. In conclusion, this study demonstrated that ANE-induced oxidative damage to isolated and cellular DNA which may result from the generation of hydrogen peroxide, and iron may serve as a catalyst in this process. Furthermore, ripe ANE generated higher oxidative DNA damage levels compared to tender ANE.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed1996

Plasmid insertional mutation may confer glucocorticoid responsiveness of cell growth.

Culture of CHO.K1, a Chinese hamster ovary cell line, requires no particular care about the glucocorticoid level in media. Cell growth of CHO.K1 is little affected by dexamethasone at concentrations up to 3 microM. A clone of CHO.K1 stably transfected with an expression vector displayed a favored growth in dexamethasone-containing media. Ironically, dexamethasone was used to trigger the expression of antisense PCNA from the expression vector to impede the cell growth. Proliferating cell nuclear antigen (PCNA), an auxiliary factor of DNA polymerase delta, is required for cell proliferation. The stable cell clone, designated as B11 had a retarded growth rate as compared to its parental cell. However, the B11 cell growth rate and the cell cycle progression were increased by dexamethasone. The glucocorticoid produced no similar effect on the parental cell or other stable transfectants of the same plasmid. Thus, the stimulatory effect of dexamethasone on B11 has little connection with the expression of antisense PCNA and possibly involves a relevant gene in the B11 genome that was mutated due to the random plasmid insertion. A preliminary effort in identifying the targeted gene was made by using plasmid rescue method, and two plasmids were obtained. The rescued DNA of both plasmids specifically hybridized genomic DNA of the parental cells, and one of these plasmids detected a cellular transcript that was absent in B11 cells, suggesting its potential for further investigation.

Animals↗

PubMed1996

Role of oxidative DNA damage in hydroxychavicol-induced genotoxicity.

Chewing betel quid has been linked to the development of oral cancer. In Taiwan, fresh Piper betle inflorescence is uniquely added to betel quid, and hydroxychavicol is the major phenolic components of P.betle inflorescence. In this study, we tested the mutagenic potential of hydroxychavicol in Salmonella typhimurium TA97, TA98, TA100 and TA102 with and without Aroclor-1254 induced S9 fraction. The results showed that hydroxychavicol was positive in S.typhimurium TA102 without metabolic activation. This increase in revertants was partially inhibited by catalase and superoxide dismutase. In Chinese hamster ovary (CHO-K1) cells, hydroxychavicol induced chromosome aberrations in a dose-dependent manner (10-50 microM) and the majority were chromosome-type aberrations. Hydroxychavicol also significantly increased the frequency of micronuclei in CHO-K1 cells up to 3-fold at a concentration of 40 microM. In addition, hydroxychavicol dose-dependently (0.1-20 microM) induced copper-dependent strand breaks in plasmid DNA. We further tested the oxidative DNA damage potential of hydroxychavicol by measuring 8-hydroxydeoxyguanosine (8-OH-dG) formation in CHO-K1 cells following an 18-h incubation and found that hydroxychavicol (6.25-100 microM) induced 8-OH-dG levels dose-dependently. The increase of 8-OH-dG formation was positively correlated (r = 0.79) with the hydroxychavicol-induced cytotoxicity. In conclusion, hydroxychavicol may exert its genotoxic potential through oxidative DNA damage.

8-Hydroxy-2'-Deoxyguanosine↗

PubMed1996

Epsilon 4 allele of apolipoprotein E increases risk of Alzheimer's disease in a Chinese population.

We examined the apolipoprotein E genotype in 56 Chinese patients with late-onset sporadic Alzheimer's disease (AD) and 57 Chinese control subjects of similar age. The frequency of epsilon 4 in the AD group was significantly higher than that in the control group (23.2% versus 7.9%, p = 0.003). The odds ratio for AD in individuals with either one or two epsilon 4 was 2.96 (95% CI 1.11 to 8.03). The linear trend for AD in proportion to alleles of epsilon 4 was also significant (chi 2 = 8.2, p = 0.004). Our results support the association between epsilon 4 and AD in the Chinese.

Age of Onset↗

PubMed1996

Serum interleukin-6 levels reflect disease status of gastric cancer.

OBJECTIVES: Interleukin-6 (IL-6) has been shown to be associated with cancer development. However, its role in gastric cancer patients has never been investigated. Our intent was to investigate this role. METHODS: Using enzyme-linked immunosorbent assay, we determined the IL-6 levels in sera of patients with gastric cancer (n = 218), benign gastric lesions (n = 9), and hepatitis B (n = 9), and in normal subjects (n = 85). C-reactive protein (CRP) levels and Helicobacter pylori antibodies were simultaneously determined in 150 and 133 gastric cancer patients, respectively. Serial serum IL-6 levels were also measured in 14 gastric cancer patients. Six gastric cancer cell lines were examined for IL-6 secretion. For comparison, serum carcinoembryonic antigen levels in all gastric cancer patients were also determined. RESULTS: Mean IL-6 levels were significantly higher in patients with gastric cancer (10.0 +/- 9.5 pg/ml) than in patients with benign gastric lesions (2.6 +/- 0.5 pg/ml), in hepatitis B carriers (2.8 +/- 0.9 pg/ml), and in normal subjects (2.5 +/- 0.3 pg/ml). Gastric cancer patients with IL-6 > 10 pg/ml increased in a stage-related manner (p = 0.0001). Serum IL-6 levels increased when four patients had recurrence, whereas IL-6 levels decreased in 10 patients with a disease-free status after gastric resection. Serum IL-6 levels correlated moderately with serum CRP levels (r = 0.5, p < 0.01), but not with H. pylori antibody titers (p = 0.23). Serum IL-6 levels correlated with survival, but not as an independent prognostic indicator. Conversely, patients with CEA > 5 ng/ml increased in a stage-related manner, but this is not an effective reflection of disease progression. Five of six gastric cancer cell lines secrete IL-6. CONCLUSIONS: Serum IL-6 levels correlate with disease status of gastric cancer and may be used as a new tumor marker for monitoring treatment and response of gastric cancer patients.

Adenocarcinoma↗

PubMed1995

Non-destructive analysis of the protein conformational structure of human pituitary adenomas using reflectance FT-IR microspectroscopy.

Reflectance Fourier-transform infrared microspectroscopy was used to determine the secondary structure and composition of the different human pituitary adenomas. Non-functioning pituitary adenomas exhibited similar protein secondary structure and conformational composition, but active pituitary adenomas revealed different behavior. The differences in secondary structure for the different human pituitary adenomas might possibly be due to the different protein conformations of the proliferated adenoma tissues and various hormones shared.

Adenoma↗

PubMed1995

Mechanistic study of the inhibition of aflatoxin b1-induced hepatotoxicity by dimethyl 4,4'-dimethoxy-5,6,5',6'-dimethylenedioxy biphenyl-2, 2'-dicarboxylate.

The mechanism of DDB (dimethyl 4,4'-dimethoxy-5,6,5',6'-dimethylenedioxy biphenyl-2,2'-dicarboxylate) prevention of aflatoxin B1 (AFB1)-induced hepatotoxicity in rats has been investigated. Pretreatment of DDB (200 mg/kg) daily for 4 days significantly suppressed (P < 0.05) the AFB1-induced hepatic damage as evidenced by the increase of serum marker enzymes. DDB induced rat hepatic cytochrome P450IA1, IIB1 and glutathione S-transferase activities. The hepatic microsomes derived from DDB treated rats increased the mutation frequency of AFB1 and enhanced the binding of AFB1 to DNA. However, the hepatic S9 fraction from DDB treated rats showed a protective effect against AFB1-induced damage. It is concluded that the protective effect of DDB against AFB1-induced damage might be mediated by the induced glutathione S-transferase activity and not from the accelerated hepatic cytochrome P450 detoxification pathway of AFB1 which was previously believed.

Aflatoxin B1↗