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D Tang

Publications and source records attributed to D Tang.

At least 91 records · Page 5Linked to original sources
PubMed1998

Expression of functional domain of chicken gizzard calponin.

A full-length cDNA of the function domain of wild-type chicken gizzard calponin was cloned into expression vector pAED4 and the recombinant function domain of wild-type calponin was expressed in an Escherichia coli expression system. The actin domain of calponin (CaP-B) can bind with actin and it is a requisite for its inhibition of ATPase and vasoconstriction of smooth muscle. In this study, the cDNA of CaP-B was inserted into vector pAED4 by direction-cloning method. The cDNA of CaP-B was obtained with PCR cloning technique. The recombinant DNA pAED4-Cap-B was transformed into E. coli BL21 (DE3) and identified with the restriction analysis. The bacterial clones containing transformants were induced to be highly expressed in E. coli BL21 (DE3). The target protein was detected and identified by Western Blot analysis. The content of target protein was as high as 10% of the whole protein after overnight (16 h) culture. The results confirmed that Cap-B was relatively highly expressed in E. coli.

Animals↗
PubMed1998

The intermediate filament protein peripherin is a marker for cerebellar climbing fibres.

Immunocytochemical staining with antibodies to the class III intermediate filament protein peripherin reveals discrete subpopulations of neurons and nerve fibres throughout the rat central nervous system. Some of these fibres enter the cerebellar granular and molecular layers. Here we use light and electron microscopic immunocytochemistry and confocal fluorescence microscopy to identify the peripherin positive fibres in the molecular layer of the cerebella of various mammals. (1) The peripherin positive fibres in the molecular layer have morphological attributes of climbing fibres, and peripherin positive fibres are also detected in the olivo-cerebellar tract. Furthermore peripherin positive neurons can be seen in the inferior olive, from which climbing fibres originate. (2) The peripherin positive molecular layer fibres rapidly degenerate in rats treated with 3-acetylpyridine (3-AP), a reagent which destroys neurons in the inferior olive, and the time course of degeneration of these mirrors that previously described for 3-AP induced destruction of climbing fibres. (3) Cerebella of other mammal species tested (mouse, rabbit, pig, cow and human) revealed a similar peripherin staining pattern in the cerebellum, including fibres in the molecular layer with the morphology of climbing fibres. (4) We also noted peripherin positive spinocerebellar and vestibulocerebellar mossy fibres in the cerebellar granular layer of folia known to receive these inputs. (5) A subset of perivascular nerve fibres are also peripherin positive. These results show that peripherin is a useful marker for mammalian cerebellar climbing fibres, and that a subset of morphologically distinct cerebellar mossy fibres are also peripherin positive.

Animals↗
PubMed1998

[Influence of dietary habits and body weight on blood uric acid in the elderly].

Influence of dietary habits, body weight on blood uric acid was studied in 416 elderly people. The result showed that level of blood uric acid in the people who had habits of drinking alcohol, tea and taking hot foods was higher than that who never had those habits (P < 0.05 or 0.01). It also showed that level of blood uric acid was significantly increased in the over-weight or obesity people (P < 0.05). The hyperuricemia incidence in the over-weight or obesity people is 27.4 per cent, and it is 2 times and 3.4 times of the people with ideal weight and weak-weight, respectively. It is suggested that the patients with gout or hyperuricemia give up drinking alcohol, tea and taking hot foods for their health. Reducing body weight is one of the effective measures to prevent and treat gout or hyperuricemia in the elderly.

Aged↗
PubMed1998

Detection of gsp oncogene in growth hormone-secreting pituitary adenomas and the study of clinical characteristics of acromegalic patients with gsp-positive pituitary tumors.

OBJECTIVE: To investigate the incidence and clinical characteristics of gsp oncogene positive growth hormone-secreting adenomas of Chinese acromegalic patients. METHODS: Continuously 40 patients were studied. Serum hormone levels of pituitary and target glands were measured and growth hormone (GH)-TRH stimulating tests were done before transsphenoidal or transfrontal hypophysectomy. Deoxyribonucleic acid (DNA) was extracted from the frozen tumor tissue, and the DNA fragment encompassing codon 201 and 227 of the Gs alpha gene was amplified by polymerase chain reaction (PCR). Point mutations at codon 201 and 227 were detected using PCR direct sequencing method in order to get the incidence of gsp oncogene in GH secreting adenomas. RESULTS: Of 40 tumors studied, 22 (55%) were gsp positive. The point mutation from CGT (Arg) to TGT (Cys) at codon 201 was detected in 21 pituitary tumors, but the point mutation from CAG (Gln) to CTG (Leu) at codon 227 of the Gs alpha gene was found in only 1 tumor. All of the point mutations are heterozygous. The number of gsp positive patients which have 30% or more decrease of serum GH concentration after glucose inhibition is less than that of gsp negative patients (P = 0.042). Compared to gsp negative patients, most of gsp positive patients showed paradoxical response to TRH stimulation (P = 0.002). There were more gsp positive patients with the tumor diameter less than 25 mm (P = 0.029) and with normal GH levels in OGTT after surgery (P = 0.007). CONCLUSIONS: Gsp mutation is one of the major intrinsic defects in the pathogenesis of growth hormone-secreting pituitary tumors and the identification of gsp mutation can be a reference for classification and prognosis of GH tumors.

Acromegaly↗
PubMed1998

[Glucose-6-phosphate dehydrogenase mutations among Cantonese revealed by polymerase chain reaction using dried blood spots].

OBJECTIVE: To analyze G6PD gene mutation in 168 Cantonese G6PD deficient male infants. METHODS: PCR products were amplified directly from dried blood spots on filter paper using 7 pairs of special primers followed by digestion with a restriction enzyme. RESULTS: Of the 168 samples, 72(42.8%) were G6PD 1376 G-->T mutation, 35 (20.8%) were G6PD 1388 G-->A, 30(17.9%) were G6PD 95 AG, 6(3.6%) were G6PD 392 G-->T, and 3(1.8%) were G6PD 1024 C-->T. No G6PD 493 A-->G and 487 G-->A mutation were found, and 22(13.1%) were not defined. CONCLUSION: 1. The three G6PD mutations at 1376, 1388 and 95 were common in Cantonese. 2. Dried blood spots collected on filter paper provide an easy way of sample collection, storage and transport for the epidemiological study of inherited disease.

Adult↗
PubMed1998

[Determination of vincristini sulfas in tumor cells by high performance liquid chromatography].

Vincristini sulfas (VCR) is an important common antitumor drug. The resistance to anticancer drug is the main cause of the chemotherapy failure. To screen the drugs which can reverse VCR resistance for VCR resistant cell strain, an analytical method has been established for the determination of VCR concentration in tumor cells using high performance liquid chromatography (HPLC). The stainless steel column was 25 cm x 4.6 mm i.d. packed with totally porous, spherical silica particles (5 microns). A solution of methanol and 0.02 mol/L dipotassium hydrogen phosphate (80:20, V/V) adjusted to pH 6.6 with H3PO4 was employed as the mobile phase. The flow rate was 1 mL/min. Chromatography was performed with ultraviolet detector at 267 nm. The method is simple, rapid and selective. Linear calibration curve for VCR was measured within the range of 10 to 200 mg/L with correlation coefficient of 0.999 8. The lowest detection limit was 4 mg/L tumor cells extract. The HPLC method described is suitable for clinical monitoring and pharmacokinetic study of VGR.

Antineoplastic Agents, Phytogenic↗
PubMed1998

[The measurement of baroreflex sensitivity in stress-induced hypertensive rats by spectral analysis].

Simultaneous spectral analysis of short-term systolic blood pressure variability (SBPV) and heart rate variability (HRV) was applied to determine the change of baroreflex sensitivity (BRS) in Sprague-Dawley (SD) rats. The modulus of the transfer function between fluctuations in systolic pressure and beat-to-beat interval would be an appropriate quantification of BRS. The animal experiment was performed on two groups of rats: normotensive rats and stress-induced hypertensive rats (SIHR). V1-receptor antagonist d(CH2)5Tye(Me) AVP of arginine vasopressin (AVP) was microinjected into intracerebroventricle. The results showed that, in base-line condition before administration, although the BRS in very low frequency band (0-0.035 Cycle/Beat, VLF), low frequency band (0.035-0.12 Cycle/Beat, LF), high frequency band (0.12-0.32 Cycle/Beat, HF) and total BRS(the sum of the three bands) were all decreased, the BRS in VLF(P < 0.05) and LF(P < 0.01) decreased significantly by statistics; and that, to the SIHR, the BRS in VLF was significantly lower after AVP-V1 administration than in base-line condition, while to normotensive rats, the BRS did not change. It indicates that the facilitating effect of AVP on the beroreflex in SIHR is mainly due to V1-receptor in central nerve system. In summary, the transfer function between SBPV and beat-to-beat interval fluctuation could be an index of BRS. This method can be developed for future clinical application.

Animals↗
PubMed1998

Pancreatic adenocarcinomas with DNA replication errors (RER+) are associated with wild-type K-ras and characteristic histopathology. Poor differentiation, a syncytial growth pattern, and pushing borders suggest RER+.

The clinical and pathological features of carcinomas of the pancreas with DNA replication errors (RER+) have not been characterized. Eighty-two xenografted carcinomas of the pancreas were screened for DNA replication errors using polymerase chain reaction amplification of microsatellite markers. Cases with microsatellite instability in at least two markers of a minimum of five tested were considered RER+. RER status was correlated with histological appearance, karyotype of the carcinomas when available, K-ras mutational status, and patient outcome. Three (3.7%) of the eighty-two carcinomas were RER+. In contrast to typical gland-forming adenocarcinomas of the pancreas, all three RER+ carcinomas were poorly differentiated and had expanding borders and a prominent syncytial growth pattern. Neither a Crohn's-like lymphoid infiltrate nor extracellular mucin production were prominent. Ductal adenocarcinomas of the pancreas typically contain a mutant K-ras gene, yet all three RER+ carcinomas had wild-type K-ras. One of the three RER+ carcinomas was karyotyped and showed a near diploid pattern. All three of the RER+ tumors were removed via Whipple resection. One of the three patients is free of disease 16 months after pancreaticoduodenectomy, one is alive and free of tumor at 52 months but developed two colon carcinomas during this period, and the third died of pancreatic cancer at 4 months. None of the three patients had a family history of colorectal carcinoma. A review of the K-ras wild-type carcinomas in a previously characterized series of pancreatic carcinomas with known K-ras mutational status identified two additional cancers with poor differentiation, a syncytial growth pattern, and pushing borders. Both of the cancers were diploid and both patients were longterm survivors (over 5 years). The inclusion of such patients in previous prognostic studies of pancreas cancer may explain the failure of histological grade to be a predictor of prognosis. These data suggest that DNA replication errors occur in a small percentage of resected carcinomas of the pancreas and that wild-type K-ras gene status and a medullary phenotype characterized by poor differentiation, and expanding pattern of invasion, and syncytial growth should suggest the possibility of DNA replication errors in carcinomas of the pancreas.

Adenocarcinoma↗
PubMed1998

In vivo radioprotective effects of angiogenic growth factors on the small bowel of C3H mice.

This study was undertaken to determine if acidic or basic fibroblast growth factor (FGF1 or FGF2) or vascular endothelial growth factor (VEGF) alters the radiation response of small bowel after total-body irradiation (TBI). Female C3H mice were treated with various doses of angiogenic growth factor administered intravenously 24 h before or 1 h after TBI. Radiation doses ranged from 7 to 18 Gy. End points measured were the number of crypts in three portions of the small bowel, the frequency of apoptosis of crypt cells at various times after TBI, and the LD50/30 (bone marrow syndrome) and LD50/6 (GI syndrome). Fibroblast growth factors alone, without TBI, decreased the number of crypts per circumference significantly. Among the factors tested, FGF2 caused the greatest decline in baseline crypt number. Despite this decrease in the baseline crypt number, after irradiation the number of surviving crypts was greater in animals treated with growth factor. The greatest radioprotection occurred at intermediate doses of growth factor (6 to 18 pg/mouse). Mice treated with FGF1 and FGF2 had crypt survival curves with a slope that was more shallow than that for saline-treated animals, indicating radiation resistance of crypt stem cells in FGF-treated mice. The LD50/6 was increased by approximately 10% for all treatments with angiogenic growth factors, whether given before or after TBI. Apoptosis of crypt cells was maximum at 4 to 8 h after TBI. The cumulative apoptosis was decreased significantly in animals treated with angiogenic growth factors, and the greatest protection against apoptosis was seen in animals treated with FGF2 prior to TBI. All three angiogenic growth factors tested were radioprotective in small bowel whether given 24 h before or 1 h after irradiation. The mechanism of protection is unlikely to involve proliferation of crypt stem cells, but probably does involve prevention of radiation-induced apoptosis or enhanced repair of DNA damage of crypt cells.

Angiogenesis Inducing Agents↗
PubMed1997

Induction of the cholesterol metabolic pathway regulates the farnesylation of RAS in embryonic chick heart cells: a new role for ras in regulating the expression of muscarinic receptors and G proteins.

We propose a novel mechanism for the regulation of the processing of Ras and demonstrate a new function for Ras in regulating the expression of cardiac autonomic receptors and their associated G proteins. We have demonstrated previously that induction of endogenous cholesterol synthesis in cultured cardiac myocytes resulted in a coordinated increase in expression of muscarinic receptors, the G protein alpha-subunit, G-alphai2, and the inward rectifying K+ channel, GIRK1. These changes in gene expression were associated with a marked increase in the response of heart cells to parasympathetic stimulation. In this study, we demonstrate that the induction of the cholesterol metabolic pathway regulates Ras processing and that Ras regulates expression of G-alphai2. We show that in primary cultured myocytes most of the RAS is localized to the cytoplasm in an unfarnesylated form. Induction of the cholesterol metabolic pathway results in increased farnesylation and membrane association of RAS. Studies of Ras mutants expressed in cultured heart cells demonstrate that activation of Ras by induction of the cholesterol metabolic pathway results in increased expression of G-alphai2 mRNA. Hence farnesylation of Ras is a regulatable process that plays a novel role in the control of second messenger pathways.

Animals↗
PubMed1997

Mefloquine compared with doxycycline for the prophylaxis of malaria in Indonesian soldiers. A randomized, double-blind, placebo-controlled trial.

BACKGROUND: Mefloquine and doxycycline are the two drugs recommended for prophylaxis of malaria for visitors to areas where Plasmodium falciparum is resistant to chloroquine. OBJECTIVE: To compare the efficacy and tolerability of mefloquine with those of doxycycline as prophylaxis for malaria. DESIGN: Randomized, double-blind, placebo-controlled field trial of chemoprophylaxis of malaria. SETTING: Northeastern Irian Jaya, Indonesia. PARTICIPANTS: 204 Indonesian soldiers. INTERVENTION: After radical curative treatment, participants were randomly assigned to receive 100 mg of doxycycline per day and mefloquine placebo; 250 mg of mefloquine per week (preceded by a loading dose of 250 mg/d for 3 days) and doxycycline placebo; or placebos for both drugs. Prophylaxis lasted approximately 13 weeks. MEASUREMENTS: The primary end point for efficacy was the first occurrence of malaria, as documented by a positive malaria smear. Malaria smears were obtained weekly and when patients had symptoms suggesting malaria. Reported symptoms were recorded daily, and an exit study questionnaire was conducted. RESULTS: In the placebo group, 53 of 69 soldiers developed malaria (9.1 person-years), resulting in an attack rate of 5.8 cases per person-year (95% CI, 4.3 to 7.7 cases per person-year). Plasmodium falciparum accounted for 57% of cases, and P. vivax accounted for 43% of cases. No malaria occurred in the 68 soldiers (16.9 person-years) in the mefloquine group; thus, the protective efficacy of mefloquine was 100% (CI, 96% to 100%). In the doxycycline group, P. falciparum malaria occurred in 1 of 67 soldiers (16.0 person-years), yielding a protective efficacy of 99% (CI, 94% to 100%). Both drugs were very well tolerated. CONCLUSIONS: Mefloquine and doxycycline were both highly efficacious and well tolerated as prophylaxis of malaria in Indonesian soldiers.

Adult↗
PubMed1997

Cyclin-dependent kinase 5 (Cdk5) activation domain of neuronal Cdk5 activator. Evidence of the existence of cyclin fold in neuronal Cdk5a activator.

Neuronal Cdk5 activator (Nck5a) differs from other cyclin-dependent kinase (Cdk) activators in that its amino acid sequence is only marginally similar to the cyclin consensus sequence. Nevertheless, computer modeling has suggested that Nck5a contains the cyclin-fold motif recently identified in the crystal structure of cyclin A. In the present study, a number of truncation mutants and substitution mutants of the Nck5a were produced and tested for the Cdk5 activation and Cdk5 binding activity. The active domain of Nck5a determined by using the truncation mutants consists of the region spanning residues 150 to 291. The size of Nck5a active domain is essentially the same as that of cyclin A required for Cdk2 activation (Lees, E. M., and Harlow, E. (1993) Mol. Cell. Biol. 13, 1194-1201). The change, or the lack of change, in Cdk5 activation activity observed with a number of substitution mutants may be understood on the basis of structure and function relationship of cyclin A. These results provide support to the previous suggestion (Brown, N. R., Noble, M. E. M., Endicott, J. A., Garman, E. F., Wakatsuki, S., Mitchell, E., Rasmussen, B., Hunt, T., and Johnson, L. N. (1995) Structure 3, 1235-1247) that the activation domain of Nck5a adopts a conformation similar to that of cyclin A. They also provide a partial answer to the question of how Nck5a, a non-cyclin, activates a cyclin-dependent kinase.

Amino Acid Sequence↗
PubMed1997

The study on the relationship between the expression of calponin and gallstone formation.

In order to understand the molecular mechanisms of gallstone formation, the expression of calponin in animal model of gallstone disease was studied. High-cholesterol diet was given to the guinea pigs to induce gallstone formation. RT-PCR and Western-blotting were used to evaluate expression level of calponin gene. Down-regulation of calponin gene expression was observed in animals with gallstone, while myosin expression was relatively stable. Our results indicated that the decrease of calponin could increase the pressure of Oddi's sphincter, aggravate the stasis of bile and promote the gallstone formation.

Animals↗
PubMed1997

Changes in the expression of novel Cdk5 activator messenger RNA (p39nck5ai mRNA) during rat brain development.

We previously reported that a neuron-specific Cdk5 activator, p35nck5ai, was most prominent in the newborn rat brain. In the adult brain, the expression decreased in most regions except hippocampus and primary olfactory cortex. A novel neuron-specific Cdk5 activator, p39nck5ai, has been recently cloned. To clarify whether two activators were differentially distributed throughout brain development, in this study, we examined the spatial and temporal expression of p39nck5ai in the development rat brain. Northern blot analysis showed that p39nck5ai expression was low in 15-day old fetuses and newborn, and was most prominent in the 1-3 week-old rat brains. In the adult rat brain, expression declined to the same level as in newborn rat brain. In situ hybridization showed that p39nck5ai mRNA was weakly expressed in all neurons of all regions in the newborn rat brain and the transcriptional level was highest in all regions in the 3 week-old rat brain. In the adult, expression was decreased in most neurons except Purkinje and granule cells in the cerebellum which retained high levels. These results suggest that p35nck5a and p39nck5ai may have different functional roles in distinct brain regions during different states of the rat brain development.

Aging↗
PubMed1997

Affinity selection and mass spectrometry-based strategies to identify lead compounds in combinatorial libraries.

The screening of diverse libraries of small molecules created by combinatorial synthetic methods is a recent development which has the potential to accelerate the identification of lead compounds in drug discovery. We have developed a direct and rapid method to identify lead compounds in libraries involving affinity selection and mass spectrometry. In our strategy, the receptor or target molecule of interest is used to isolate the active components from the library physically, followed by direct structural identification of the active compounds bound to the target molecule by mass spectrometry. In a drug design strategy, structurally diverse libraries can be used for the initial identification of lead compounds. Once lead compounds have been identified, libraries containing compounds chemically similar to the lead compound can be generated and used to optimize the binding characteristics. These strategies have also been adopted for more detailed studies of protein-ligand interactions.

Binding, Competitive↗
PubMed1997

Characterization of additional casein kinase I sites in the C-terminal "tail" region of chicken and rat neurofilament-M.

In previous studies we have identified Ser502, Ser528, and Ser534 as target sites in chicken neurofilament middle molecular mass protein (NF-M) for casein kinase I (CKI) in vitro and have shown that these sites are also phosphorylated in vivo. We now make use of a combination of molecular biological and protein chemical techniques to show that two additional in vivo phosphorylation sites in chicken NF-M, Ser464 and Ser471, can also be phosphorylated by CKI in vitro. These two sites are conserved in higher vertebrate NF-M molecules, and recombinant protein constructs containing the homologous rat NF-M peptides can be phosphorylated by CKI in vitro, suggesting that phosphorylation of these sites is conserved at least in higher vertebrates. The two new sites are adjacent to a conserved peptide sequence (VEEIIEET-V) found once in higher vertebrate NF-M molecules and twice in lamprey NF-180. Variants of this sequence are also found in neurofilament low and high molecular mass proteins (NF-L and NF-H) and alpha-internexin, and in mammalian NF-L are known to be associated with in vivo phosphorylation sites. We speculate that CKI phosphorylation in general, and these sites in particular, may be important in neurofilament function.

Amino Acid Sequence↗
PubMed1997

Variability in PAH-DNA adduct measurements in peripheral mononuclear cells: implications for quantitative cancer risk assessment.

Biomarkers such as DNA adducts have significant potential to improve quantitative risk assessment by characterizing individual differences in metabolism of genotoxins and DNA repair and accounting for some of the factors that could affect interindividual variation in cancer risk. Inherent uncertainty in laboratory measurements and within-person variability of DNA adduct levels over time are putatively unrelated to cancer risk and should be subtracted from observed variation to better estimate interindividual variability of response to carcinogen exposure. A total of 41 volunteers, both smokers and nonsmokers, were asked to provide a peripheral blood sample every 3 weeks for several months in order to specifically assess intraindividual variability of polycyclic aromatic hydrocarbon (PAH)-DNA adduct levels. The intraindividual variance in PAH-DNA adduct levels, together with measurement uncertainty (laboratory variability and unaccounted for differences in exposure), constituted roughly 30% of the overall variance. An estimated 70% of the total variance was contributed by interindividual variability and is probably representative of the true biologic variability of response to carcinogenic exposure in lymphocytes. The estimated interindividual variability in DNA damage after subtracting intraindividual variability and measurement uncertainty was 24-fold. Inter-individual variance was higher (52-fold) in persons who constitutively lack the Glutathione S-Transferase M1 (GSTM1) gene which is important in the detoxification pathway of PAH. Risk assessment models that do not consider the variability of susceptibility to DNA damage following carcinogen exposure may underestimate risks to the general population, especially for those people who are most vulnerable.

Adult↗