[Research progress of hepatic non-parenchymal cells in nonalcoholic steatohepatitis].

Non-alcoholic steatohepatitis (NASH) is an important link for the progression of metabolic-related fatty liver disease to end-stage liver disease such as cirrhosis and hepatocellular carcinoma, which seriously endangers human health. NASH pathogenesis is complex, and involves the interaction between hepatic parenchymal and non-parenchymal cells (NPCs), sinusoidal endothelial cells, Kupffer cells, hepatic stellate cells, and so on. Herein, the relevant research progress of NPCs in the pathogenesis of NASH is reviewed in order to further understand the role of NPCs in NASH.

Correlation between TRAIL and caspase-8 expression and their relationship with cell proliferation and apoptosis in human osteosarcoma.

Osteosarcoma is a common malignant bone tumor that mainly affects children and adolescents. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor superfamily. Caspase-8 appears in the upstream of apoptosis signaling pathway among caspases. We investigated TRAIL and caspase-8 levels in osteosarcoma patients to determine their correlation with cell proliferation and apoptosis. Osteosarcoma and osteochondroma patients receiving surgery in our hospital were selected. TRAIL and caspase-8 expression levels in tissue were determined by immunohistochemistry, and protein levels in cells were evaluated by western blotting. Human osteosarcoma cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The osteosarcoma and osteochondroma cell cycles and apoptosis were investigated by flow cytometry. Correlation analysis was applied to TRAIL and caspase-8 levels during cell apoptosis. Positive TRAIL and caspase-8 expression rates in osteosarcoma tissue were significantly lower than in the controls (P < 0.05). TRAIL (0.114 ± 0.002) and caspase-8 (0.352 ± 0.124) levels in experimental cells were obviously lower than in the controls (P < 0.05). Osteosarcoma cells in the experimental group demonstrated higher proliferation and lower apoptosis at 24, 48, and 72 h (P < 0.05). The experimental cell number increased in the G1 stage and decreased in the S stage (P < 0.05). TRAIL and caspase-8 proteins showed positive correlation with apoptosis in osteosarcoma (P < 0.05). Human osteosarcoma presented reduced TRAIL and caspase-8 levels with enhanced cell proliferation and reduced apoptosis. TRAIL and caspase-8 expression levels were positively correlated with apoptosis in osteosarcoma.

Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus samples obtained from farms in Gansu, China.

Porcine epidemic diarrhea poses significant sanitation problems in the porcine industry, and has negatively affected the economy in recent years. In this study, 48 fecal specimens were collected from piglets from four intensive swine farms located in the Gansu Province of China. The molecular diversity and phylogenetic relationships between porcine epidemic diarrhea viruses (PEDV) prevalent in Gansu were probed, and the resultant proteins were characterized. Sequence analysis of the spike protein (S) genes showed that each specimen had unique characteristics, and that the PEDV1/S/4 strain could be differentiated from the others via a unique mutation of the S gene. The phylogeny of S glycoprotein showed that all strains were clustered into two major groups. The four Gansu PEDV field strains were characterized into different groups; this finding was consistent with the results of the protein characterization prediction. This analysis additionally revealed the unique characteristics of each specimen. The results of this study could be used to elucidate the prevalence of PEDV and contribute to the prevention of PEDV in Gansu.

Detecting bit-flip errors in a logical qubit using stabilizer measurements.

Quantum data are susceptible to decoherence induced by the environment and to errors in the hardware processing it. A future fault-tolerant quantum computer will use quantum error correction to actively protect against both. In the smallest error correction codes, the information in one logical qubit is encoded in a two-dimensional subspace of a larger Hilbert space of multiple physical qubits. For each code, a set of non-demolition multi-qubit measurements, termed stabilizers, can discretize and signal physical qubit errors without collapsing the encoded information. Here using a five-qubit superconducting processor, we realize the two parity measurements comprising the stabilizers of the three-qubit repetition code protecting one logical qubit from physical bit-flip errors. While increased physical qubit coherence times and shorter quantum error correction blocks are required to actively safeguard the quantum information, this demonstration is a critical step towards larger codes based on multiple parity measurements.

Production of coloured glass-ceramics from incinerator ash using thermal plasma technology.

Incineration is a major treatment process for municipal solid waste in Taiwan. It is estimated that over 1.5 Mt of incinerator ash are produced annually. This study proposes using thermal plasma technology to treat incinerator ash. Sintered glass-ceramics were produced using quenched vitrified slag with colouring agents added. The experimental results showed that the major crystalline phases developed in the sintered glass-ceramics were gehlenite and wollastonite, but many other secondary phases also appeared depending on the colouring agents added. The physical/mechanical properties, chemical resistance and toxicity characteristic leaching procedure of the coloured glass-ceramics were satisfactory. The glass-ceramic products obtained from incinerator ash treated with thermal plasma technology have great potential for building applications.

The activity in ex vivo expansion of cord blood myeloid progenitor cells before and after cryopreservation.

A total of 50 human umbilical cord blood (UCB) samples were studied. The hematopoietic stem/progenitor (CD34+) populations were isolated from UCB mononuclear cells (MNC) by means of immunomagnetic separation. Double immunofluorescent staining of UCB CD34+ cells revealed that there was a high proportion (82.33 +/- 4.47%) of CD34+ cells co-expressing CD13, while the percentage of CD34+ CD33+ cells was much lower (22.17 +/- 3.35%). In contrast, for co-expressing lymphoid differentiation antigens, the proportion of CD34+CD38+ cells (38.34 +/- 6.09%) was relatively higher than that of CD34+CD10+ cells (11.52 +/- 1.24%) or CD34+CD2+ cells (9.84 +/- 2.30%). For stimulating the ex vivo expansion of UCB progenitor cells, no single hematopoietic growth factor (HGF) was efficacious when used alone, while combination of 4 HGFs, such as GM-CSF, G-CSF, IL-3, and SCF could induce a 55-fold increase in the myeloid progenitor cells, day-14 CFU-GM, in a short term of 7 days' liquid culture. Cryopreservation of UCB as MNC preparations at -196 degrees C could satisfactorily retain the number and activity of CD34+ cells. After thawing, a high recovery rate of about 80% CD34+ cells was obtained. When suspended in liquid cultures containing a combination of 4 HGFs, as shown above, the frozen cord blood progenitor cells could be well expanded, reaching a >50-fold increase in day-14 CFU-GM, which was very similar to that of the fresh UCB samples. In addition, a similar result was also seen in CFU-GEMM, indicating that after cryopreservation the recovered UCB progenitor cells retain an intact clonogeneic ability capable of efficiently responding to hematopoietic growth factors for ex vivo expansion.

[Metabolic calculation of the growth phase in rHSA fermentation].

The model equations of the growth phase of rHSA fermentation were derived on the base of both elemental balance and metabolic balance. The unknown parameters of the model were estimated by multivariable optimization. The model can preferably describe the relations between different macroscopic reaction rates of the process and provide the key for the high-density cultivation of Pichia pastoris.

Free fatty acid fractions from some vegetable oils exhibit reduced survival time-shortening activity in stroke-prone spontaneously hypertensive rats.

Previously, we demonstrated that several vegetable oils that included low-erucic rapeseed oil markedly shortened the survival time (by approximately 40%) of stroke-prone spontaneously hypertensive (SHRSP) rats as compared with perilla oil, soybean oil, and fish oil. We considered that a factor other than fatty acids is toxic to SHRSP rats, because the survival time-shortening activity could not be accounted for by the fatty acid compositions of these oils. In fact, a free fatty acid (FFA) fraction derived from lipase-treated rapeseed oil was found to be essentially devoid of such activity. A high-oleate safflower oil/safflower oil/perilla oil mixture exhibited a survival time-shortening activity comparable to that of rapeseed oil, but the activity of this mixed oil was also reduced by lipase treatment. A partially hydrogenated soybean oil shortened the survival time by approximately 40%, but a FFA fraction derived from lipase-treated partially hydrogenated soybean oil shortened it by 13% compared with soybean oil. Fatty acid compositions of the rapeseed oil and a FFA fraction derived from lipase-treated rapeseed oil were similar, but those of hepatic phospholipids of rats fed the oil and FFA were slightly but significantly different. These results support the interpretation that the survival time-shortening activity exhibited by some vegetable oils is due to minor components other than fatty acids, and that an active component(s) were produced in or contaminated soybean oil during the partial hydrogenation processes.

Protein carbonyl content roughly reflects the unsaturation of lipids in muscle but not in other tissues of stroke-prone spontaneously hypertensive strain (SHRSP) rats fed different fats and oils.

We examined in vivo the effect of dietary fats and oils with different peroxidizability on protein carbonyl content, the presumed index of free radical-mediated protein oxidation. For 15.6 months, SHRSP (stroke-prone spontaneously hypertensive strain) rats were fed a diet supplemented with lard, safflower oil, perilla oil or fish oil/soybean oil, the peroxidizability of which increases in this order. The peroxidizability of tissue lipids was positively correlated with the protein carbonyl content in skeletal muscle, but not in the brain, heart or liver. The protein carbonyl content in the lard group was higher in the brain and liver compared to the other dietary groups. These results contradict the concept that long-term feeding of easily autoxidizable fatty acids allows the accumulation of lipid peroxides to accelerate the development of the free radical diseases, and suggest that tissue protein carbonyl content is not a simple reflection of autoxidizability-related lipid peroxidation but is also influenced by other biochemical processes.

Unusual effects of some vegetable oils on the survival time of stroke-prone spontaneously hypertensive rats.

Preliminary experiments have shown that a diet containing 10% rapeseed oil (low-erucic acid) markedly shortens the survival time of stroke-prone spontaneously hypertensive (SHRSP) rats under 1% NaCl loading as compared with diets containing perilla oil or soybean oil. High-oleate safflower oil and high-oleate sunflower oil were found to have survival time-shortening activities comparable to that of rapeseed oil; olive oil had slightly less activity. A mixture was made of soybean oil, perilla oil, and triolein partially purified from high-oleate sunflower oil to adjust the fatty acid composition to that of rapeseed oil. The survival time of this triolein/mixed oil group was between those of the rapeseed oil and soybean oil groups. When 1% NaCl was replaced with tap water, the survival time was prolonged by approximately 80%. Under these conditions, the rapeseed oil and evening primrose oil shortened the survival time by approximately 40% as compared with n-3 fatty acid-rich perilla and fish oil; lard, soybean oil, and safflower oil with relatively high n-6/n-3 ratios shortened the survival time by roughly 10%. The observed unusual survival time-shortening activities of some vegetable oils (rapeseed, high-oleate safflower, high-oleate sunflower, olive, and evening primrose oil) may not be due to their unique fatty acid compositions, but these results suggest that these vegetable oils contain factor(s) which are detrimental to SHRSP rats.

Complete amino acid sequence of an acidic, cardiotoxic phospholipase A2 from the venom of Ophiophagus hannah (King Cobra): a novel cobra venom enzyme with "pancreatic loop".

A phospholipase A2 (OHV A-PLA2) from the venom of Ophiophagus hannah (King cobra) is an acidic protein exhibiting cardiotoxicity, myotoxicity, and antiplatelet activity. The complete amino acid sequence of OHV A-PLA2 has been determined using a combination of Edman degradation and mass spectrometric techniques. OHV A-PLA2 is composed of a single chain of 124 amino acid residues with 14 cysteines and a calculated molecular weight of 13719 Da. It contains the loop of residues (62-66) found in pancreatic PLA2s and hence belongs to class IB enzymes. This pancreatic loop is between two proline residues (Pro 59 and Pro 68) and contains several hydrophilic amino acids (Ser and Asp). This region has high degree of conformational flexibility and is on the surface of the molecule, and hence it may be a potential protein-protein interaction site. A relatively low sequence homology is found between OHV A-PLA2 and other known cardiotoxic PLA2s, and hence a contiguous segment could not be identified as a site responsible for the cardiotoxic activity.

Role of enzymatic activity in the antiplatelet effects of a phospholipase A2 from Ophiophagus hannah snake venom.

A phospholipase A2 (OHV A-PLA2) from the venom of Ophiophagus hannah (King cobra) is an acidic protein exhibiting antiplatelet activity. In in vitro tests, OHV APLA2 showed a marked inhibitory effect on platelet aggregation induced by ADP, collagen and arachidonic acid in both human whole blood and platelet-rich plasma in a dose-dependent manner. The antiplatelet effects of OHV A-PLA2 did not increase when preincubation times of platelets and OHV A-PLA2 were prolonged indicating phospholipid hydrolysis did not significantly contribute to the antiplatelet effects. Alkylation of active site His residue using p-bromophenacyl bromide resulted in the complete loss of enzymatic activity, but the modified enzyme retained more than 30% of its antiplatelet effects. These results indicate that the antiplatelet effects of OHV A-PLA2 appear to be independent of its enzymatic activity, and there are separate sites responsible for the catalytic and antiplatelet activities.

Effect of captopril on intracellular pH in vascular smooth muscle cells.

AIM: To observe the effect of captopril (Cap) on intracellular pH (pHi) in aortic smooth muscle cells (ASMC). METHODS: Cultured ASMC derived from rat and rabbit aortae were loaded with the fluorescent dye BCECF and pHi was determined using digital image processing method. RESULTS: The pHi of untreated SHR and WKY rats were 7.37 +/- 0.29 and 7.19 +/- 0.31, respectively. Oral Cap decreased pHi (7.11 +/- 0.26, P < 0.05) and exaggerated pHi response to angiotensin II (Ang-II, 0.1 mumol.L-1) in ASMC of SHR rats vs WKY rats (0.14 +/- 0.05 vs 0.21 +/- 0.05 pH units, P < 0.01). Cap in vitro had no effect on Ang-II induced intracellular alkalinization in ASMC of rabbits. CONCLUSION: Oral Cap inhibits Na+(-)H+ exchange activity in ASMC of SHR rats.

Effect of rapeseed and dietary oils on the mean survival time of stroke-prone spontaneously hypertensive rats.

Low-fat conventional diets supplemented with 5 or 10% vegetable oils were fed to stroke-prone spontaneously hypertensive rats (SHR-SP) from weaning and the mean survival times were determined. A 1% aqueous sodium chloride solution was used as drinking water throughout the experiments. In four separate experiments, the rapeseed oil group showed a significantly shorter mean survival time. The relative mean survival times were 50-59% (rapeseed oil group), 78-100% (soybean oil group) and 86% (microbial oil group) as compared with the group fed perilla oil (100%). The group which received 4-fold diluted rapeseed oil exhibited a significantly shorter survival time as compared with the group receiving soybean oil. Although the feeding experiments were performed under very simple and restricted conditions, these results suggest that the rapeseed oil prepared for human use contains a factor (s) which is toxic to SHR-SP rats.

Effects of dietary vegetable oils on behavior and drug responses in mice.

Previously, we noted significant differences in the behavioral patterns of mice fed safflower oil with a very low alpha-linolenate/linoleate ratio and perilla oil with a high alpha-linolenate/linoleate ratio from mothers to offsprings. In this report, we compared the behavior and drug responses in mice fed diets containing six different vegetable oils-corn, rapeseed, soybean, safflower, perilla and a mixture of perilla and safflower oils- for a relatively short period: 8 months after weaning. Soybean oil is a component of most conventional diets and was used as a control. The alpha-linolenate/linoleate ratios of the oils appeared to affect the locomotor activities in a wheel cage: the activity decreased in the order of safflower, the mixture (1:1) and the perilla oil groups. However, the rapeseed oil group exhibited much higher locomotor activity than that expected from the alpha-linolenate/linoleate ratio. Additionally, the rapeseed oil group exhibited unusual behavior patterns, including higher ambulation and rearing activities, faster acquisition of the water maze task and slower habituation behavior as compared with the control group. Susceptibility to pentobarbital anesthesia tended to be higher in the rapeseed oil group. The differences in the alpha-linolenate/linoleate ratios of these oils alone do not account for the observed differences in the behavioral patterns among the six dietary groups. Although we cannot exclude the possibility that the observed behavioral anomaly is due to the unique fatty acid composition of rapeseed oil, we speculate that a factor(s) other than fatty acids in rapeseed oil affected nervous system functions.

Effects of captopril and enalapril on intracellular Ca2+ in vascular smooth muscle cell.

AIM: To determine whether angiotensin-converting enzyme inhibitors can affect Ca2+ handling in cultured aortic smooth muscle cells (ASMC) directly. METHODS: Cultured ASMC derived from rat aorta were loaded with the intracellular Ca2+ ([Ca]2+i) fluorescent indicator Fura 2-AM and digital image processing technique was used. RESULTS: Resting [Ca2+]i was greater in ASMC from SHR vs WKY (P < 0.01). KCl-, norepinephrine (NE)-, and angiotensin II (Ang)-induced [Ca2+]i increases were enhanced in ASMC of SHR vs WKY (220 +/- 6, 212 +/- 8, and 215 +/- 14 vs 199 +/- 6, 202 +/- 7, and 195 +/- 7 nmol.L-1, respectively). Captopril (Cap) and enalapril (Ena) had no inhibitory effect on KCl-, NE-, and Ang-induced [Ca2+]i increases in ASMC of WKY. Cap and Ena inhibited KCl-, NE-, and Ang-increased [Ca2+]i in ASMC of SHR (210 +/- 7, 194 +/- 6, and 201 +/- 6 nmol.L-1, respectively). Ena and nifedipine similarly decreased KCl-, NE-, and Ang-increased [Ca2+]i. CONCLUSION: Cap blocked KCl-, NE-, and Ang-increased ([Ca2+]i) via a voltage-dependent Ca2+ channel of which function and specificity was altered in ASMC of SHR.

Pathological changes induced by an acidic phospholipase A2 from Ophiophagus hannah venom on heart and skeletal muscle of mice after systemic injection.

An acidic phospholipase A2 (OHV A-PLA2) isolated from the venom of the king cobra (Ophiophagus hannah) was tested for its ability to cause pathological changes to myocardium, skeletal muscle and cardiac ganglia. White mice were injected intravenously with dose of 8 mg/kg or 4 mg/kg of OHV A-PLA2 and tissue samples were taken at 6 or 24 hr. Light microscopic examination failed to show significant changes in cardiac muscle and ganglia. Skeletal muscle showed myofibre degeneration and necrosis. Electron microscopic study revealed myodegeneration in cardiac and skeletal muscles, and reduction in synaptic vesicle population of preganglionic nerve terminals in cardiac ganglia. Ultrastructural changes in tissues were dose related. The lower dose (4 mg/kg) of OHV A-PLA2 produced mild myocardial changes, the myofilaments were intact but contracted, and the A band and I band were skewed. OHV A-PLA2 caused myocardial degeneration at a higher dose of 8 mg/kg. The changes included dissolution of actin and myosin filaments, dilatation and disorganization of sarcoplasmic reticulum and degeneration of mitochondria. The skeletal muscle lesions were more severe than the myocardial changes. Some of the myofibrils were severely disorganized and lack typical striated appearance, sarcomeres disrupted, most of mitochondria were vesiculated and destroyed.

Light-emitting aeromonas and plesiomonas generated by transconjugating luxAB from Escherichia coli.

The transposon derivative has been placed on a transposition suicide vector to yield pDB30 in Escherichia coli WA803. A simple method, using a Tn5 derivative Tn5-Lux, has been successfully devised for the introduction and stable expression of the bioluminescence property in Pseudomonas sp., Agrobacterium sp., and Rhizobium sp. In this study, there was also successful mating between Escherichia coli WA803(pDB30) and strains of Acromonas hydrophila and Plesiomonas shigelloides. These bacteria emitted bioluminescence after they gained pDB30 by transconjugation.

[Assessment of a new four-hour diagnostic kit--RapID onE system for the identification of enteric bacteria].

RapID onE System is a newly developed four-hour rapid diagnostic kit for the identification of enteric bacteria. To know the effectiveness of this system, we used 125 strains of oxidase-negative, gram-negative bacilli for this evaluation. Except for Acinetobacter calcoaceticus, all the bacilli belong to family Enterobacteriaceae. The bacterial strains of this assessment belong to 12 genera and 20 species. Among them, 84 strains were freshly isolated from clinical specimens and 41 strains were frozen (-70 degrees C) stock clinical isolates. The results show that 115 (92.0%) strains were correctly identifed to the species level. It yielded 92.9% and 90.2% of correct identification of fresh isolates and frozen stocks, respectively. In this paper, the reading criteria of RapID onE System would also be discussed.