Search PubMed⌕ Search

Biomedical subjects

J H Li

Publications and source records attributed to J H Li.

At least 55 records · Page 3Linked to original sources
PubMed2000

Folk medicine Terminalia catappa and its major tannin component, punicalagin, are effective against bleomycin-induced genotoxicity in Chinese hamster ovary cells.

Terminalia catappa L. is a popular folk medicine for preventing hepatoma and treating hepatitis in Taiwan. In this paper, we examined the protective effects of T. catappa leaf water extract (TCE) and its major tannin component, punicalagin, on bleomycin-induced genotoxicity in cultured Chinese hamster ovary cells. Pre-treatment with TCE or punicalagin prevented bleomycin-induced hgprt gene mutations and DNA strand breaks. TCE and punicalagin suppressed the generation of bleomycin-induced intracellular free radicals, identified as superoxides and hydrogen peroxides. The effectiveness of TCE and punicalagin against bleomycin-induced genotoxicity could be, at least in part, due to their antioxidative potentials.

Animals↗
PubMed2000

Properties of Ca(2+) release-activated Ca(2+) channel block by 5-nitro-2-(3-phenylpropylamino)-benzoic acid in Jurkat cells.

Ca(2+) release-activated Ca(2+) current (I(crac)) has been previously characterized biophysically in Jurkat lymphocytes and other non-excitable cells, but pharmacology remains poorly developed. The present objective was to delineate with whole cell recording details of the interaction of the chloride channel blocker, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), with I(crac) in Jurkat cells. NPPB reversibly inhibited I(crac) in a concentration-dependent manner (IC(50)=5 microM). Kinetics for block and unblock of I(crac) by NPPB indicated a bimolecular interaction. Michaelis-Menten analysis indicated that NPPB interacts competitively with extracellular Ca(2+) permeating the I(crac) pathway. Finally, analysis of the pH dependence of I(crac) block by NPPB revealed a reduction in the apparent affinity during extracellular alkalinization that based on the pK(a) for NPPB, suggested that the neutral form of NPPB blocks the Ca(2+) release-activated Ca(2+) (CRAC) channel. Taken together, our results suggest a direct interaction between NPPB and the CRAC channel, and should help guide insights for developing novel and more selective analogues.

Calcium↗
PubMed2000

Tumor immunity in perforin-deficient mice: a role for CD95 (Fas/APO-1).

CTL and NK cells use two distinct cytocidal pathways: 1) perforin and granzyme based and 2) CD95L/CD95 mediated. The former requires perforin expression by the effectors (CTL or NK), whereas the latter requires CD95 (Fas/APO-1) expression by the target. We have investigated how these two factors contribute to tumor immune surveillance by studying the immunity of perforin-deficient mice against the progressor C57BL/6 Lewis lung carcinoma 3LL, which expresses no CD95 when cultured in vitro. Unexpectedly, the results indicated that the perforin-independent CD95L/CD95 pathway of CTL/NK plays a role in acting against D122 and Kb39.5 (39.5) high and low metastatic sublines, respectively, derived from the 3LL tumor. Although no membrane-bound CD95 was detected on cultured D122 and 39. 5 cells, surface CD95 expression on both D122 and 39.5 was considerably up-regulated when the tumors were grown in vivo. A similarly enhanced expression of CD95 was observed with three additional tumors; LF-, BW, and P815, injected into syngeneic and allogeneic mice. The finding of up-regulated CD95 expression on tumor cells placed in vivo suggests that a CD95-based mechanism plays a role in tumor immunity at early stages of tumor growth. Consequently, the progressive down-regulation of CD95 expression during tumor progression may indeed be an escape mechanism as previously reported. Together, these results suggest a role for CD95-dependent, perforin-independent immunity against certain tumors.

Animals↗
PubMed2000

An electron microscopic-cytochemical localization of plasma membrane Ca(2+)-ATPase activity in poplar apical bud cells during the induction of dormancy by short-day photoperiods.

Plasma membrane (PM) Ca2+-ATPase activity in poplar apical bud meristematic cells during short-day (SD)-induced dormancy development was examined by a cerium precipitation EM-cytochemical method. Ca2+-ATPase activity, indicated by the status of cerium phosphate precipitated grains, was localized mainly on the interior face (cytoplasmic side) of the PM when plants were grown under long days and reached a deep dormancy. A few reaction products were also observed on the nuclear envelope. When plant buds were developing dormancy after 28 to 42 d of SD exposure, almost no reaction products were present on the interior face of the PM. In contrast, a large number of cerium phosphate precipitated grains were distributed on the exterior face of the PM. After 70 d of SD exposure, when buds had developed a deep dormancy, the reaction products of Ca2+-ATPase activity again appeared on the interior face of the PM. The results seemed suggesting that two kinds of Ca2+-ATPases may be present on the PM during the SD-induced dormancy in poplar. One is the Ca2+-pumping ATPase, which is located on the interior face of the PM, for maintaining and restoring the Ca2+ homeostasis. The other might be an ecto-Ca2+-ATPase, which is located on the exterior face of the PM, for the exocytosis of cell wall materials as suggested by the fact of the cell wall thickening during the dormancy development in poplar.

Calcium-Transporting ATPases↗
PubMed2000

[Modulation of epidermal growth factor on the synthesis of pulmonary surfactant and its mechanism].

AIM: To study the effect of EGF at physiological level on PS synthesis on cultured lung explants without serum. METHODS: The total phospholipids and major phospholipid components in lung tissues were determined. The amount of 3H-choline incorporation into PC was measured. RESULTS: 1. 10(-9) mol/L EGF enhanced 3H-choline incorporation into PC at 8 h and reached the maximum at 16 h. 2. The synthesis of total phospholipids, PC and PG increased from lung tissues exposed to 10(-9) mol/L EGF, but membrane characteristic phospholipid showed no change. 3. TP 47, a PTK inhibitor and H7, a PKC inhibitor could block EGF-induced PS synthesis. The release of LDH from lung explants could not be changed by EGF, TP 47 and H7. CONCLUSION: EGF at physiological level could enhance the adult rat PS synthesis, and its modulated mechanisms involved PTK and PKC pathways.

Animals↗
PubMed2000

[Changes of the responses of arterioles to noradrenaline, acetylcholine and nitroglycerin after limb ischemia-reperfusion].

OBJECTIVE: To explore the mechanism of microvascular spasm after limb ischemia-reperfusion. METHODS: The rabbit hindlimb normothermic tourniquet ischemia model was employed. The tendon on the dorsum of the foot was exposed for observation of microvessels. The responses of arterioles on tendon surface to topical application of 10(-6) mol/L noradrenaline (NE) (a vasoconstrictor), 10(-6) mol/l acetylcholine(Ach) (an endothelium-dependent vasodilator) and 10(-4) mol/L nitroglycerin(NTG) (an endothelium-independent vasodilator) were observed at the period of ischemia and following 30 minutes of reperfusion after 2 hours and 5 hours of ischemia by use of intravital microscopy. RESULTS: No significant changes in the responses of arterioles to NE, Ach and NTG were noted following 30 minutes of reperfusion after 2 hours of ischemia compared with pre-ischemia. The constrictor responses of arterioles to NE were still not significantly altered following 30 minutes of reperfusion after 5 hours of ischemia, however, the dilation responses to Ach and NTG were significantly decreased (to Ach P < 0.01; to NTG, P < 0.05). CONCLUSION: Reperfusion after 5 hours of ischemia significantly impairs both the endothelium-dependent and endothelium-independent vasodilation, meanwhile preserves constrictor responses to NE, these may contribute to the genesis of the vasospasm in ischemia reperfusion.

Acetylcholine↗
PubMed2000

[Molecular phylogeny of Giardia lamblia based on triose phosphate isomerase (tim) gene sequence].

OBJECTIVE: To investigate the intraspecies phylogeny and genetic diversity of G. lamblia. METHODS: Total genomic DNA was extracted and partial fragments of tim gene were amplified by PCR. All the nucleotide sequences were analyzed by means of phylogenetic analysis, which were constructed with pasimony and N-J methods. RESULTS: A total of 124 variable sites (23% of the total sequences detected) were defined, most of which were found at the silent sites of codons. Two similar phylogenetic trees were constructed, subdividing 16 isolates into two groups. CONCLUSION: The genetic diversity of G. lamblia appear to be little affected by both the host and geography, while natural-selection might play an important role at the DNA molecular evolution level of tim gene. The tim gene may be considered to be a very useful genetic marker of the population genetic structure of G. lamblia.

Animals↗
PubMed1999

Determination of tramadol by capillary gas chromatography with flame ionization detection. Application to human and rabbit pharmacokinetic studies.

A rapid, sensitive, precise and accurate capillary gas chromatographic assay with flame ionization detection was developed for the determination of tramadol in human, rabbit, pig and dog plasma. It is comprised of only a one-step extraction procedure with dichloromethane at pH 11.15 and gas chromatography on a capillary column. The recoveries of tramadol and meperidine (internal standard) were greater than 88%. Calibration graphs were linear over the concentration range 12.5-10,000 ng/ml with a coefficient of variation, both within-day and between-day, of less than 10% at any level. The limit of detection was 8 ng/ml of plasma based on signal-to-noise ratio of 3. Six other clinically used analgesics were investigated to check for potential interferences and their analytical conditions. The specificity of this assay was checked with two major metabolites of tramadol (M1: O-demethyltramadol; M2: N-demethyltramadol). Tramadol in plasma did not decompose significantly at -20 degrees C for 56 days. Pharmacokinetic application with intravenous tramadol in humans and rabbits revealed that tramadol followed a two-compartment open model with one distribution phase and one elimination phase. The distribution and elimination half-lives in humans were 1.02 and 141.9 min. The distribution and elimination half-lives in rabbits were 7.31 and 63.2 min, respectively.

Analgesics, Opioid↗
PubMed1999

The effects of combining ionizing radiation and adenoviral p53 therapy in nasopharyngeal carcinoma.

PURPOSE: Nasopharyngeal carcinoma (NPC) is a malignant disease of the head/neck region, with a 5-year survival level of approximately 65%. To explore gene therapy as a novel approach which might improve outcome, we have shown previously that introduction of human recombinant wild-type p53 mediated by the adenoviral vector (Ad5CMV-p53) was cytotoxic in two human nasopharyngeal carcinoma (NPC) cell lines (CNE-1 and CNE-2Z). The current work was designed to determine whether this strategy, combined with ionizing radiation (XRT), was more effective than either treatment alone. METHODS AND MATERIALS: CNE-1, CNE-2Z, and a normal human nasopharyngeal fibroblast strain, KS1, were infected with 2- and 6-plaque-forming units (pfu)/cell of Ad5CMV-p53, respectively. These doses were isoeffective for beta-galactosidase activity in the CNE-1 and CNE-2Z cells. XRT was administered 24 h post-infection, and Western blot analyses were conducted for p53, p21WAF1/CIP1, bax, and bcl-2 2 days after XRT. Cell survival was assessed using a clonogenic assay. Presence of DNA ladders reflecting apoptosis was detected using DNA agarose gel electrophoresis, and cell cycle was analyzed using flow cytometry. RESULTS: The combination of Ad5CMV-p53 plus XRT (2, 4, and 6 Gy) resulted in an approximately 1-log greater level of cytotoxicity compared to that observed with XRT alone for both NPC cell lines. The two modalities appear to be interacting in a synergistic manner in cancer cells, but not in KS1 fibroblasts. XRT alone stimulated minimal p53 expression in control cells; Ad5CMV-p53 alone induced significant recombinant p53 expression, which was not further enhanced by the addition of XRT. Similar observations were made for p21WAF1/CIP1 expression. No changes were observed for bax or bcl-2 expression with any of these treatments. Apoptosis was induced following 4 Gy of XRT alone, but was observed after only 2 Gy when combined with Ad5CMV-p53. Cell cycle analysis indicated that Ad5CMV-p53 infection did not perturb the cell cycle beyond that observed with XRT alone. CONCLUSION: p53 gene therapy and XRT appears to interact in a synergistic manner; underscoring the significant potential of this novel strategy in the treatment of NPC.

Adenoviridae↗
PubMed1999

Genetic polymorphisms of orosomucoid on the Han population in Nanjing of China.

Human orosomucoid (ORM) is a major binding protein for various basic drugs. The genetic polymorphisms of ORM could be responsible for interindividual variation in the plasma binding of basic drugs, which might influence their effects or toxicities. The genetic polymorphisms of ORM on the Han population in Nanjing of China were analyzed by isoelectric focusing (IEF) on polyacrylamide gels following by immunoblotting. After desialylation of sera from 220 unrelated Chinese subjects, the band patterns of ORM showed that the polymorphism of the structural locus ORM1 is controlled by three codominant autosomal alleles, ORM1*F1, ORM1*F2 and ORM1*S, which presented five phenotypes, ORM1 F1, ORM1 S, ORM1 F1F2, ORM1 F1S, and ORM1 F2S. The allele frequencies were: ORM1*F1=0.7068, ORM1*F2=0. 0182, ORM1*S=0.2750. The results presented in this paper indicate the ORM1 locus is polymorphic and the ORM2 locus is monomorphic in sera from the Han population in Nanjing of China.

Alleles↗
PubMed1999

Role of TYR70 in the N-glycosidase activity of neo-trichosanthin.

Trichosanthin (TCS) is a type I ribosome-inactivating protein (RIP) which possesses rRNA N-glycosidase activity. TCS has long been used as an abortifacient in China. In recent years, its immunomodulatory, anti-tumor and anti-HIV properties have attracted more and more attention. An isoform of trichosanthin, neo-trichosanthin (n-TCS), has been cloned and expressed as recombinant protein. The biochemical studies revealed that n-TCS has virtually the same rRNA N-glycosidase activity as TCS. The crystal structure of n-TCS is similar to TCS. The crystal of Y70A n-TCS, the mutant of recombinant n-TCS, was soaked in sodium citrate buffer (pH 5.5) containing 25% KCl and AMP (10 mg/ml) prior to data collection. After structure determination and refinement, no electron density corresponding to adenine can be detected around the active pocket. Furthermore, the reaction products of Y70A n-TCS and AMP incubated at various reaction times were analyzed using HPLC. No adenine can be detected. These results suggest that Tyr70 is crucial to n-TCS for its substrate recognition, binding and perhaps N-glycosidase activity.

Adenine↗
PubMed1999

Neurokinin B receptor (NK3)-containing neurons in the paraventricular and supraoptic nuclei of the rat hypothalamus synthesize vasopressin and express Fos following intravenous injection of hypertonic saline.

Subnuclear localization of neurokinin B receptor (NK3) in the paraventricular and supraoptic nuclei of the hypothalamus was immunohistochemically investigated in the rat. In the paraventricular hypothalamic nucleus, intense neurokinin B receptor-like immunoreactivity was found in the posterior magnocellular part, moderate to weak neurokinin B receptor-like immunoreactivity was seen in the other parts. In the supraoptic nucleus, intense neurokinin B receptor-like immunoreactivity was distributed in its principal part, and a few neurons with neurokinin B receptor-like immunoreactivity were found in the retrochiasmatic part. Co-localization of neurokinin B receptor-like immunoreactivity with vasopressin-like immunoreactivity was examined through serial adjacent sections. Neurons with both neurokinin B receptor-like immunoreactivity and vasopressin-like immunoreactivity were primarily found in the supraoptic nucleus and posterior magnocellular part of the pavaventricular nucleus. A small number of neurons with neurokinin B receptor-like immunoreactivity and vasopressin-like immunoreactivity were also seen in the circular nucleus and the region surrounding blood vessel in the anterior hypothalamus. Many neurokinin B receptor-containing neurons in the paraventricular and supraoptic nuclei, as well as in circular nucleus and the region surrounding the blood vessel, expressed Fos-like immunoreactivity after intravenous injection of hypertonic saline. The present study demonstrated that a large proportion of neurokinin B receptor-like immunoreactive neurons in the paraventricular hypothalamic and supraoptic nuclei contained vasopressin-like immunoreactivity, and expressed Fos-like immunoreactivity after intravenous administration of hypertonic saline. The results suggest that neurokinin B receptor in the two nuclei may be involved in modulation of the release of vasopressin when the internal environment is disturbed.

Animals↗
PubMed1999

Inhibition of protein kinase C activity promotes heat-induced apoptosis in RIF-1 but not in TR-4 cells.

PURPOSE: Inhibition of protein kinase C (PKC) activity has been demonstrated to reduce thermotolerance (TT), presumably by decreasing heat shock protein (HSP) production. Therefore, the interest was in evaluating this relationship further in two isogenic murine tumour cell lines: RIF-1 and its thermoresistant TR-4 selectant. MATERIALS AND METHODS: TT was induced in RIF-1 and TR-4 cells (45 degrees C for 15 min, then 37 degrees C for 6 h) with or without Ro31-8220, a specific inhibitor of PKC. PKC activity was assayed by determining the catalytic transfer of ATP to a specific substrate peptide. Survival was determined using the clonogenic assay. Apoptosis was quantitated by counting the number of cells demonstrating apoptosis after staining with acridine-orange/ ethidium bromide. Production of the inducible form of HSP70 was assessed using Western blot. RESULTS: At 2 microM Ro31-8220, >80% of PKC activity was inhibited in both cell lines, which was associated with no cytotoxicity at 37 degrees C. Basal HSP70 level was approximately 10-fold higher in the TR-4 compared with the RIF-1 cells. Upon TT induction, HSP70 level increased significantly in both cell lines, which was suppressed in the presence of Ro31-8220, but the relative amount of HSP70 remained high in the TR-4 cells. At 24 h, heat-induced apoptosis increased from 4 to 38% in RIF-1 cells in the presence of Ro31-8220, which was associated with a 26% reduction in clonogenic survival after thermotolerant heating. In contrast, <1% of TR-4 cells demonstrated apoptosis even with the highest dose of Ro31-8220, and no effect on survival was observed. CONCLUSION: Inhibition of PKC activity reduces HSP70 induction, which in turn is associated with promotion of heat-induced apoptosis in RIF-1 cells. However, the survival signals in the TR-4 cells are so strong, that even 80% inhibition of PKC activity has minimal impact on heat-induced apoptosis and survival in this thermoresistant cell line.

Animals↗
PubMed1999

Genetic toxicity of cocaine.

Cocaine is a widely abused drug. Recently, it has been shown to induce teratogenesis in both humans and animals. Cocaine-induced teratogenicity has been associated with reactive oxygen species (ROS), which are generated by cytochrome P450 during cocaine biotransformation. Since ROS have been reported to induce genotoxicity, it is of interest to know whether cocaine and/or its metabolites are also genotoxic. In this study, Chinese hamster ovary K1 cells were employed as a model system to investigate the genetic toxicity of cocaine in the presence or absence of rat liver S9 fraction. Cocaine-induced cytotoxicity was potentiated when S9 was present, indicating the cytochrome P450 metabolism plays a role in cocaine-mediated cytotoxicity. Cocaine treatments per se induced a few chromosome aberrations while treatments of cocaine plus S9 caused a significant increase in chromosome aberrations. In contrast, cocaine induced micronuclei (MN) formation and hypoxanthine-guanine phosphoribosyltransferase mutation only in the presence of S9. Therefore, cocaine itself is at best a weak clastogen, whereas metabolite(s) of cocaine is/are truly inducer(s) of clastogenesis and mutagenesis. Cocaine treatments alone also induced a significant increase in sister chromatid exchange frequency but the addition of S9 did not affect the results. Free radical scavengers, including superoxide dismutase and catalase, efficiently decreased the frequency of cocaine plus S9-induced MN, implying that ROS are indeed important components in cocaine-induced genotoxicity. The observation that non-toxic doses of cocaine can inhibit intercellular metabolic cooperation suggests that cocaine may also be a tumor promoter. Our data supports that cocaine could possess genotoxicity in addition to its well-known neurotoxicity and teratogenicity.

Animals↗
PubMed1999

Role of Arg163 in the N-glycosidase activity of neo-trichosanthin.

Three mutant crystals of neo-trichosanthin (n-TCS), R163K, R163H and R163Q, were obtained by the hanging drop vapor diffusion method. Structure determination indicated that there are no significant differences between the mutants and n-TCS except in the active pocket. All of them were also soaked in sodium citrate buffer (pH 4. 5) containing 20% KCl and 10 mg/ml AMP. Structure determination suggests that in the active pocket of the crystals of R163K and R163H, parallel to the aromatic ring of Tyr70, each mutant possesses an adenine. The relationship between structure and function is discussed. Biochemical analysis reveals that the mutants R163K and R163H have N-glycosidase activity, while R163Q does not. This suggests that R163 is a crucial residue for the enzyme activity of n-TCS, and its role is providing proton.

Adenosine Monophosphate↗
PubMed1999

Resonant activations for a fluctuating barrier system driven by dichotomous noise and Gaussian white noise.

We consider the escape over a fluctuating barrier in the presence of a dichotomous noise and a Gaussian white noise. It is shown that the mean first passage time (MFPT) over the fluctuating barrier displays two resonant activations. One is the resonant activation of the MFPT as a function of the flipping rate of the fluctuating potential barrier; the other is the resonant activation of the MFPT as a function of the transition rate of the dichotomous noise. In addition, we find that the dichotomous noise can weaken the former resonant activation, but enhance the latter one. By further study, we find that, when the fluctuating potential barrier is driven by two or more dichotomous noises, there are three or more resonant activations.

Journal Article↗