[Clinical characteristics of patients with delayed hepatolenticular degeneration].

Objective: To evaluate the clinical manifestations, metal metabolism, imaging characteristics and treatment response in patients with delayed Wilson disease (WD). Methods: Patients with untreated WD (40 with delayed onset and 40 with non-delayed onset) were enrolled. Twenty healthy people were included as normal controls. All patients were evaluated with modified Young scale neural symptom scores, grade of Child liver function and mental symptoms rating scale, magnetic resonance imaging (MRI) scan, magnetic sensitive imaging (susceptibility weighted imaging, SWI), metal metabolism. Corrected phase (CP) was measured at SWI. After 2 week treatment, neurologic symptoms, liver function, and metal metabolism were reviewed. Results: The total score of neurological symptoms in WD patients with delayed onset was lower than that of non-delayed onset (13.00±6.87 vs. 21.13±5.53, P=0.033). The scores of SCL-90 and HAMA depression scales in patients with delayed onset were lower than those of non-delayed onset. On T(2) weighted imaging, areas including substantia nigra and thalamus, the caudate nucleus, globus pallidus, putamen presented high signal rate in patients with delated onset than those with non-delayed (P=0.022, 0.037, 0.022, 0.037, 0.029 respectively). The SWI CP values of cangbai sphere and shell nucleus in patients with delayed onset were lower than those with non-delayed onset. Patients with delayed onset had higher urinary copper than those with non-delayed onset before and after treatment (P=0.040, 0.036). After treatment, the score of abnormal tremor and gait in patients with delayed onset was decreased (P=0.037, 0.044), while as the occurrence of neurological symptoms was increased by 10%, and the liver function level in patients with delayed WD was decreased in 3 cases. Conclusions: The brain of WD patients with delayed onset is mainly composed of metal deposits, however the cell damage is not apparent. Clinical symptoms are characterized by significant liver injury, but relatively mild neurological and psychiatric symptoms. Patients with delayed WD have higher urinary copper excretion than those with non-delayed WD. Chelating agents improves the neurological symptoms in patients with delayed onset.

[History and development of carbonization of Chinese materia medica and exploration on its mechanism of hemostatic action].

The first material recorded about hair charcoal is seen in Nei jing (Inner Canon). It has a history of over 2 000 years for the carbonization of Chinese materia medica. There were controversies on this matter and its clinical application was seldom seen and underdeveloped. After the Yuan Dynasty, the main theory of carbonic herbs for hemostasis, and keeping the nature of the medicines after carbonization was gradually formed, and physicians of generations began to conduct in-depth research. Through repeated practice and verification, people summed up the suitable species of Chinese materia medica and its principle for carbonization. The methods and degree of carbonization of Chinese materia medica are reasonably discussed, with its principle and basis for application primarily interpreted.

Oxidatively modified phosphatidylserines on the surface of apoptotic cells are essential phagocytic 'eat-me' signals: cleavage and inhibition of phagocytosis by Lp-PLA2.

Diversified anionic phospholipids, phosphatidylserines (PS), externalized to the surface of apoptotic cells are universal phagocytic signals. However, the role of major PS metabolites, such as peroxidized species of PS (PSox) and lyso-PS, in the clearance of apoptotic cells has not been rigorously evaluated. Here, we demonstrate that H2O2 was equally effective in inducing apoptosis and externalization of PS in naive HL60 cells and in cells enriched with oxidizable polyunsaturated species of PS (supplemented with linoleic acid (LA)). Despite this, the uptake of LA-supplemented cells by RAW264.7 and THP-1 macrophages was more than an order of magnitude more effective than that of naive cells. A similar stimulation of phagocytosis was observed with LA-enriched HL60 cells and Jurkat cells triggered to apoptosis with staurosporine. This was due to the presence of PSox on the surface of apoptotic LA-supplemented cells (but not of naive cells). This enhanced phagocytosis was dependent on activation of the intrinsic apoptotic pathway, as no stimulation of phagocytosis occurred in LA-enriched cells challenged with Fas antibody. Incubation of apoptotic cells with lipoprotein-associated phospholipase A2 (Lp-PLA2), a secreted enzyme with high specificity towards PSox, hydrolyzed peroxidized PS species in LA-supplemented cells resulting in the suppression of phagocytosis to the levels observed for naive cells. This suppression of phagocytosis by Lp-PLA2 was blocked by a selective inhibitor of Lp-PLA2, SB-435495. Screening of possible receptor candidates revealed the ability of several PS receptors and bridging proteins to recognize both PS and PSox, albeit with diverse selectivity. We conclude that PSox is an effective phagocytic 'eat-me' signal that participates in the engulfment of cells undergoing intrinsic apoptosis.

Bioactivities of chicken essence.

The special flavor and health effects of chicken essence are being widely accepted by people. Scientific researches are revealing its truth as a tonic food in traditional health preservation. Chicken essence has been found to possess many bioactivities including relief of stress and fatigue, amelioration of anxiety, promotion of metabolisms and post-partum lactation, improvement on hyperglycemia and hypertension, enhancement of immune, and so on. These activities of chicken essence are suggested to be related with its active components, including proteins, dipeptides (such as carnosine and anserine), polypeptides, minerals, trace elements, and multiple amino acids, and so on. Underlying mechanisms responsible for the bioactivities of chicken essence are mainly related with anti-stress, anti-oxidant, and neural regulation effects. However, the mechanisms are complicated and may be mediated via the combined actions of many active components, more than the action of 1 or 2 components alone.

[Bioreactor-landfill site for leaching solution treatment].

The study utilizes the system combined bioreactor with landfill site to treat leachate. The results show that the system helped the degradation of the organic pollutants in the leachate to be divided into two-phases, the hydrolytic fermentation and acid-production phases mainly occurred in the landfill site, the acidification rate was 40%-50%, the methane-production chiefly occurred in the bioreactor(UASB). It treated the leachate significantly and benefits the collection and utilization of methane gas. Also, the system accelerates the process of degrading municipal solid waste and stabilizing landfill site.

[Relations of chemotherapy-induced tumor necrosis to plasma platin concentration and primary tumor temperature in patients with osteosarcoma in the lower extremities treated by hyperthermic isolation limb perfusion with cisplatin].

OBJECTIVES: To study the relations of chemotherapy-induced tumor necrosis to plasma concentration of platin and primary tumor temperature in hyperthermic isolation limb perfusion (HILP) with cisplatin, and to investigate the correlation between the tumor necrosis rate and the relapse-free survival (RFS). METHODS: Fifteen patients with II B osteosarcoma in the lower extremities were treated with neoadjuvant chemotherapy. Among them, 10 were male and 5 female. The average age was 18 years (ranging from 13 to 29). Chemotherapy included a single bonus dose of HILP with cisplatin (CDDP, 15 mg/m(2)). The plasma concentration of platin and the tumor temperature in the affected limb were measured during the course of the HILP with CDDP. Limb salvage surgery was performed in 9 patients and amputation in 6. After surgery, serial sections of the excised tumor specimens were made to determine the tumor necrosis rate. All of the patients were followed up for more than 5 years. RESULTS: The highest attained tumor temperature was between 41.4 degrees C - 43.0 degrees C (mean 42.3 degrees C +/- 0.46 degrees C), and the platin concentration ranged from 4.56 microg/mL to 14.66 microg/mL (mean 8.93 microg/mL +/- 3.16 microg/mL) during the course of HILP with CDDP. Thirteen patients with a necrosis rate of over 90% were well responded, and the remaining two with a necrosis rate 87.2% and 86.3% were poor responded. Eight patients remained tumor-free for 5 years. Primary tumor temperature and platin concentration were significantly related to the tumor necrosis rate (P = 0.001 and P < 0.001, respectively). No significant correlation was noted between the long-term survival rate and the three parameters in the regression analysis. CONCLUSIONS: HILP with CDDP is useful in the treatment of primary osteosarcoma. Increased tumor necrosis ensure a successful surgical excision of tumor. HILP however is still limited to the patients who are not candidates for lime salvage surgery, because the same condition of high temperature and high concentration of platin are not applicable to distant metastasis.

[Cu, Co and Se nutritive value of compound trace elements controlled-release boluses for grazing ruminants].

Twenty cattle with the similar old and weight were randomly divided into two groups for 135 d, while twenty white goats were randomly divided into two groups for 60 d. Each group consisted of ten animals, and the ratio of male and female was same. Each of animals in one group was administered an initial bolus, used as treatment group. In the other group, animals were not administered the bolus, used as control group. In the cattle feeding experiment, five cattle of each group was supplemented feed, and the other five cattles were not supplemented feed. In the goat feeding experiment, all of goats were grazing, and not supplemented feed. At the end of goat feeding experiment, three goats of each group were randomly selected to slaughter. Liver, kidney, spleen, muscle, heart, blood and matter in the rumen were collected to assay for Cu, Co and Se. The results showed that the average daily gain of cattle supplemented and no supplemented feeds in the administered and no administered group was 483.00 g, 302.83 g, 445.84 g and 282.23 g, respectively. There had significantly difference between the administered group and no administered group(p < 0.05). The average daily gain of goats in the administered and no administered groups was 68.83 g and 50.67 g, and significant difference between the two groups (p < 0.01). Administering the boluses improved the contents of Cu, Co and Se in the tissues of goats, significantly.

[Improving memory effects of eel oil capsule on memory obstruction of mice hurt by anisodine].

The effects of eel oil capsule on memory obstruction of mice were observed in step-down test, step-through test, Y-maze test and Japanese labyrinth test. The results indicated that the eel oil capsule treating group with dosages of 0.234, 0.702 and 2.106 g/kg markedly enhance the acquirement, strengthing and reappearance of memory, having the functions of improving brain memory hurt by Anisodine.

[Modulatory effects of 17beta-estradiol on the electrical activity of subfornical organ neurons].

The effects of 17beta-estradiol (E(2) ) on electrical activity of neurons in subfornical organ (SFO) slices were examined using extracelluar recording technique. The results are as follows. (1) In 15 SFO units, a low dose of E(2) (0.1 nmol/L) applied into superfusate induced an increase in discharge rate from 3.21+/-0.37 to 6.79+/-0.71 Hz (P<0.001), whereas a high dose of E(2) (100 nmol/L ) caused a decrease in discharge rate from 3.44+/-0.40 to 1.44+/-0.36 Hz (P<0.01); (2) glutamate NMDA receptor antagonist MK-801 (50 pmol/L) blocked the excitatory effects induced by low dose of 17beta-estradiol in 7 units; (3) L-arginine (L-arg, 1 mmol/L), a physiological precursor of NO, abolished the excitatory effects induced by low dose of 17beta-estradiol in 7 units; (4) application of N-omega-nitro-L-arginine methyl ester (L-NAME, 10 mmol/L), an inhibitor of NOS, blocked the inhibitory effects induced by high dose of 17beta-estradiol in 6 units. The above results suggest that the estrogen exerts dual action on SFO neuron. E(2) at low dosage increases the discharge rate of SFO neuron, an effect which may be related to the activation of NMDA receptors, whereas E(2) at high dosage decreases the discharge rate, an effect which may be attributed to the activation of NOS with resultant production of NO.

Kinetics of selenium incorporation into tissues of female mallard ducks.

Selenium is essential for both mammalian and avian species, although its metabolism in birds has been less thoroughly studied. Little information has been available on the kinetics of selenium in birds, especially as it relates to the teratogenicity seen in waterfowl consuming excessive amounts. This study describes the pharmacokinetics of small amounts of 75Se as selenious acid injected into female mallard ducks. Labeled selenium was injected into a wing vein of restrained animals and tissues taken at five different time points up to 24 h post-injection. Selenium levels as percent of injected dose were determined in liver, kidney, heart, lung, adrenals, thyroid, spleen, pancreas, ovaries, intestine, muscle and plasma. Estimates of kinetic parameters (uptake and elimination rates, time of maximum concentration and maximum concentration) were obtained with a non-linear kinetics computer program (PCNONLIN, SCI Software, Lexington, KY). Results showed four basic patterns of distribution, uptake and elimination. Visceral tissues exhibited a triphasic pattern with a rapid rise, a decline followed by a distinctive increase in levels between the last two time points. Brain uptake was continuous over the 24 h. Plasma rose rapidly and then declined to a constant level. The ovaries as a tissue of interest relating to the teratogenic effects of selenium showed the greatest relative increase at 24 h, suggesting kinetic mechanisms consistent with a pathway that might lead to accumulation of toxic levels and teratogenic effects during embryo development.

[Expression of c-fos in spinal cord, medulla oblongata and thalamus following epicardial application of adenosine].

Effects of epicardial application of adenosine on the expression of c-fos proto-oncogene in spinal cord, medulla oblongata and thalamus were examined in 12 sinoaortic denervated and vagotomized anesthetized rats. The results showed that following epicardial application of adenosine the Fos-like protein immunoreactive (FLI) neurons were remarkably increased in the dorsal horn of T3 spinal segment, nucleus paragigantocellularis lateralis (PGL) of medulla oblongata, ventral postero-lateral thalamic nucleus (VPL), posterior thalamic nucleus (Po), parafascicular thalamic nucleus (PF) and centrolateral thalamic nucleus (CL), while the blood pressure and heart rate were not affected. Only a few FLI neurons were found in the vehicle-control rats. The results indicate that epicardial application of adenosine may activate the pain-related neurons in spinal cord, medulla oblongata and thalamus.

[Effects of selenium and DTPA on cadmium excretion in rats].

Fifty-six male wistar rats were selected to study the effects of selenium and DTPA on cadmium excretion. Cadmium levels in urine, feces and tissues of the rats were determined with graphite stove method, and their kidneys were examined pathohistologically. Results showed selenium had stronger effects on cadmium excretion via feces in rats than DTPA did, and combination of both of them showed synergistic effects on it. Also, selenium could accelerate the recovery from kidney injury caused by cadmium, and combination of both of them showed synergism on it.

[Studies of radiosensitization of hyperthermia and hypertonic treatment in HEp-2 cells].

This paper presents the effects of hyperthermia, irradiation and post-irradiation treated by hypertonic solution on HEp-2 cells, alone or combined in vitro. The results showed that when the radiation treatment with hyperthermia at 44 degrees C 15', the latter had no directly killing effect on cells, a significant radiosensitization was found. The dose response curve showed that the Dq which represents sublethal damage was decreased to 4-5 times (1.49 Gy/0.31 Gy). However, the dose of D(o) which represents potentially lethal damage had almost not changed. When cells were treated with hypertonic solution (0.3 mol/L NaCl) postirradiation, or treated by combination of hyperthermia, irradiation, and hypertonic solution, the results were near the same, as compared with combination with hyperthermia alone. It is concluded that the radiosensitization is induced by hyperthermia which can inhibit the repair of the radio-induced DNA damage by heating.

An asynchronous unfolding among molecular different regions of lobster D-glyceraldehyde-3-phosphate dehydrogenase and maltotetraose-forming amylase from an Alcaligenes sp. during guanidine denaturation.

Changes in ultraviolet absorbance and intrinsic protein fluorescence of 1,4-alpha-D-glucan maltotetrahydrolase (EC 3.2.1.60) from an Alcaligenes sp. (Gram-negative bacteria 537.1) and D-glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) have been compared with their inactivation during denaturation in guanidinium-Cl solutions. The two enzymes were completely inactivated at GuHCl concentrations less than 0.6 M and this was accompanied by marked absorbance and intrinsic fluorescence changes suggesting exposure of aromatic residues. The changes of the intrinsic fluorescence of the amylase have a relatively constant plateau in emission intensities and maxima at GuHCl concentrations from 0.8-2.0 M, similar to that of muscle GAPDH. The relative activity of the enzyme increased markedly in dilute GuHCl solutions accompanied by very little change of its intrinsic fluorescence at 8 degrees C. The kinetic decrease in emission intensities, excited respectively by 230 nm and 292 nm, was different for the two enzymes. The inactivation was a biphasic process with a fast phase faster than the unfolding rate as measured by fluorescence changes in 0.5 M GuHCl solution. Similar to the inactivation process, changes in intensity of 410 nm NAD fluorescent derivative of GAPDH which is in situ at the active site is also a biphasic process under the same condition. It appears that there may be an unfolding intermediate state of the enzymes and an asynchronous unfolding process among the different regions in the molecules during GuHCl denaturation, this may be due to differences in their flexibility.

Structure and function of the 5'-flanking sequence of the human cytosolic selenium-dependent glutathione peroxidase gene (hgpx1).

Human selenium-dependent glutathione peroxidase (hGPx1) (EC 1.11.1.9) is thought to be involved in many critical cellular functions as a result of its role in glutathione-mediated reduction of toxic peroxides, and it is implicated as a mechanism of resistance against oxygen free radicals. Previous studies have demonstrated that the gene encoding hGPx1 (hgpx1) is more highly expressed in multidrug-resistant AdrR MCF-7 human breast cancer cells than in the parental WT MCF-7 cell line. In order to further study the transcriptional regulation of hgpx1, we have cloned the genomic hgpx1 gene and determined its nucleotide sequence. The 2550-base pair (bp) 5'-flanking sequence of hgpx1 contained the terminal 511 bp of the 3' end of a previously reported rhoH12 cDNA (Yeramian, P., Chardin, P., Madaule, P., and Tavitian, A. (1987) Nucleic Acids Res. 15, 1989), a ras-related oncogene. Further downstream from rhoH12, but before the start of transcription of hgpx1, RNase protection analysis revealed a transcribed sequence of at least 270 bp which we have called mid. RNA transcripts homologous to both rhoH12 (1.8 and 1.5 kilobase pairs (kb)) and mid (1.8 kb) are also more highly expressed in AdrR MCF-7 cells than in WT MCF-7 cells. We screened an AdrR MCF-7 cDNA library with the mid sequence and isolated a partial cDNA clone which contains both mid and rhoH12 sequences and is colinear with the genomic sequence which extends from 10 bp 3' to the rhoH12 stop codon to 810 bp 5' to the start of transcription of hgpx1. The start of transcription of hgpx1 in AdrR MCF-7 cells was determined by primer extension analysis. The promoter and 2 kb of the 5'-flanking sequence of hgpx1 was fused to the bacterial chloramphenicol acetyltransferase gene (hGPx1-CAT1). Analysis of deletion constructs of hGPx1-CAT1 revealed three possible cis-acting regulatory regions. The transcriptional regulation of hgpx1 was examined using the hGPx1-CAT hybrid genes and nuclear run-on studies. We found no evidence that increased mRNA transcript formation could account for different levels of hgpx1 RNA either in different breast cancer cell lines or in response to selenium.

[Inhibitory effect of micronutrients and BHA on lung cancer induced in rats].

The combined effect of Vit. A and E, selenium, cysteine and BHA on 3MC-induced lung cancer in Wistar rats was investigated. The supplementation of these compounds at their safe doses reduced the incidence of lung cancer from 47.2% to 31.6% (P less than 0.05). The total superoxide dismutase (SOD) and Mn-SOD activities in the lung cancer tissue were much lower than those in the normal tissue (P less than 0.001). It is possible that in the early stage of carcinogenesis, this change occurs only in the limited focus and does not affect the enzyme level as a whole, so SOD assay of peripheral blood can not reflect the carcinogenesis status in the lung.

Scavenging effect of extracts of green tea and natural antioxidants on active oxygen radicals.

With the use of the spin trapping methods, the scavenging effects of the extracts of green tea and other natural foods are studied. In stimulated polymorphonuclear leukocytes (PMN) system, water extract fraction 6 (F6) from green tea and green tea polyphenols (GTP) have the strongest scavenging effect on the active oxygen radicals, much stronger than vitamin C (Vc) and vitamin E (VE). Rosemary antioxidants (RA) and Curcumin (Cur) have weaker scavenging effects than Vc, but stronger than VE. In Fenton Reaction, Cur has the strongest scavenging effect (69%) on hydroxyl radicals. In irradiation, riboflavin system F6(74%) and GTP(72%) have very strong scavenging effects that are weaker than Vc, but much stronger than VE (23%). With the use of spin probe oxymetry, the oxygen consumption in respiratory burst of stimulated PMN were measured when the antioxidants existed in these systems. The results demonstrated that these antioxidants did not affect the respiratory burst of human polymorphonuclear leukocytes stimulated with PMA.