[Research progress on factors influencing early liver dysfunction after liver transplantation].

Liver transplantation is the most effective treatment for end-stage liver disease, and early graft dysfunction often occurs after surgery. Early liver dysfunction after liver transplantation has become one of the complications after liver transplantation, which seriously affects the graft and patient survival. Therefore, reducing its occurrence can be an important means to improve the prognosis of recipients after liver transplantation. This paper discusses the research progress on the definition, influencing factors, and prognosis and prediction model in order to provide better prevention and effective reference for improving the success rate and prognosis of early liver dysfunction in recipients after liver transplantation.

Epigenetic suppression of potassium-chloride co-transporter 2 expression in inflammatory pain induced by complete Freund's adjuvant (CFA).

BACKGROUND: Multiple mechanisms contribute to the stimulus-evoked pain hypersensitivity that may be experienced after peripheral inflammation. Persistent pathological stimuli in many pain conditions affect the expression of certain genes through epigenetic alternations. The main purpose of our study was to investigate the role of epigenetic modification on potassium-chloride co-transporter 2 (KCC2) gene expression in the persistence of inflammatory pain. METHODS: Persistent inflammatory pain was induced through the injection of complete Freund's adjuvant (CFA) in the left hind paw of rats. Acetyl-histone H3 and H4 level was determined by chromatin immunoprecipitation in the spinal dorsal horn. Pain behaviour and inhibitory synaptic function of spinal cord were determined before and after CFA injection. KCC2 expression was determined by real time RT-PCR and Western blot. Intrathecal KCC2 siRNA (2 µg per 10 µL per rat) or HDAC inhibitor (10 µg per 10 µL per rat) was injected once daily for 3 days before CFA injection. RESULTS: Persistent inflammatory pain epigenetically suppressed KCC2 expression through histone deacetylase (HDAC)-mediated histone hypoacetylation, resulting in decreased inhibitory signalling efficacy. KCC2 knock-down caused by intrathecal administration of KCC2 siRNA in naïve rats reduced KCC2 expression in the spinal cord, leading to sensitized pain behaviours and impaired inhibitory synaptic transmission in their spinal cords. Moreover, intrathecal HDAC inhibitor injection in CFA rats increased KCC2 expression, partially restoring the spinal inhibitory synaptic transmission and relieving the sensitized pain behaviour. CONCLUSION: These findings suggest that the transcription of spinal KCC2 is regulated by histone acetylation epigenetically following CFA. SIGNIFICANCE: Persistent pain suppresses KCC2 expression through HDAC-mediated histone hypoacetylation and consequently impairs the inhibitory function of inhibitory interneurons. Drugs such as HDAC inhibitors that suppress the influences of persistent pain on the expression of KCC2 may serve as a novel analgesic.

Chronic intrathecal infusion of mibefradil, ethosuximide and nickel attenuates nerve ligation-induced pain in rats.

BACKGROUND: T-type Ca(2+) channels (TCC) are important for pain transmission, especially the Ca(V)3.2 subtype. In this study, we examined the effects of intrathecal TCC blockers in the L5/6 spinal nerve ligation pain rat model. METHODS: Under isoflurane anaesthesia, rats received right L5/6 spinal nerve ligation and intrathecal catheters (attached to an infusion pump) were sited. After surgery, saline, mibefradil, ethosuximide or NiCl2 were given intrathecally for seven days. The right hindpaw withdrawal thresholds to von Frey hair stimuli and withdrawal latencies to radiant heat were measured before and once daily for seven days after surgery. Double immunofluorescence and western blotting were used to examine the expression of Ca(V)3.2 in dorsal root ganglion (DRG) and spinal cord. RESULTS: On post-ligation day seven, rats receiving mibefradil, ethosuximide or NiCl2 had significant higher median withdrawal thresholds (15.0, 10.2, and 10.9 g) and latencies (8.0, 7.6 and 7.6 s) than saline-treated rats (1.6 g and 4.3 s, respectively). Ca(V)3.2 was expressed in parvalbumin(+), IB4(+), CGRP(+) and VR1(+) neurones in DRG and most neurones in spinal dorsal horn. Ca(V)3.2 was up-regulated in the right L5/6 DRG and spinal cord seven days after nerve ligation. CONCLUSIONS: In this study, we demonstrated that intrathecal TCC blockers attenuate the development of nerve injury-induced mechanical allodynia and thermal hyperalgesia. Our data suggest that continuous intrathecal infusion of TCC or Ca(V)3.2 blockers may be a promising alternative for the management of nerve injury-induced pain.

Chronic intrathecal infusion of gabapentin prevents nerve ligation-induced pain in rats.

BACKGROUND: Gabapentin is an anticonvulsant and adjuvant analgesic. It is effective in several pain studies. Neuropathic pain is the most difficult type of pain to treat. In this study, we examined if intrathecal gabapentin could prevent nerve injury-induced pain. METHODS: Under isoflurane anaesthesia, male Sprague-Dawley rats (200-250 g) underwent right L5/6 spinal nerve ligation and placement of an intrathecal catheter connected to an infusion pump. After surgery, intrathecal saline or gabapentin (20 µg h(-1)) was given for 7 days (n=8 per group). The right hind paw withdrawal threshold to von Frey filament stimuli and withdrawal latency to radiant heat were determined before (baseline) and once daily for 7 days after surgery. Haematoxylin and eosin and toluidine blue staining were used to evaluate the neurotoxicity of gabapentin (40 µg h(-1)). RESULTS: Seven days after nerve ligation, the affected paw withdrawal threshold and latency of saline-treated rats decreased from the baseline 11.7 (11.7-22.2) [median (inter-quartile range)] to 1.6 (0.9-3.2) g and 10.8 (10.5-11.2) to 4.3 (4.2-7) s, respectively. Rats receiving gabapentin (20 µg h(-1)) had higher withdrawal threshold [9.9 (9.9-19.3) g] and latency [11.5 (9.7-11.9) s] on day 7 after ligation. No obvious histopathological change or growth retardation was detected after intrathecal gabapentin (40 µg h(-1)) infusion. CONCLUSIONS: We showed a preventative effect of intrathecal gabapentin on the development of nerve injury-induced mechanical allodynia and thermal hyperalgesia. Our data suggest that continuous intrathecal gabapentin may be considered as an alternative for the prevention of nerve injury-induced pain.

A method for the fabrication of low-noise carbon fiber nanoelectrodes.

A new and facile method has been developed for the fabrication of low-noise carbon fiber microelectrodes (CFMEs) and carbon fiber nanoelectrodes (CFNEs). The carbon fiber was flame-fuse sealed in the tip of the glass capillary. The CFMEs were made by cutting the protruding carbon fiber to the desired length, and the CFNEs were achieved by etching the protruding carbon on the flame to form a nanometer-scale tip. The tip of CFNEs can be controlled within the range from 100 to 300 nm. Thus, no epoxy wax was involved in the CFMEs and CFNEs. The experimental results of inspecting CFMEs and CFNEs by scanning electron microscopy demonstrated that the surface of the electrodes and the glass/fiber interface are very smooth. Therefore, the noise caused by the glass/fiber of these electrodes is much lower than that of the electrodes fabricated conventionlly. The electrodes were characterized by ferricyanide, catecholamine (dopamine,DA), norepinephrine (NE), and epinephrine (E)) and 5-hydroxytryptamine (5-HT) neurotransmitters using CV, LSV, DPV, and FSCV. The results showed that the CFMEs and CFNEs have very excellent electrochemical behavior and high sensitivity. The CV and DPV detection limits of DA, NE, and E are 7.6 x 10(-8), 7.0 x 10(-8), and 5.0 x 10(-8) mol/L, and the DPV detection limits of DA, NE, and E are 4.0 x 10(-8), 1.0 x 10(-7), and 2.2 x 10(-7) mol/L, respectively. This experiment offers a new and facile method for the fabrication of CFMEs and CFNEs of very high sensitivity and low noise.

Intrathecal magnesium sulfate attenuates algogenic behavior and spinal amino acids release after kainic acid receptor activation in rats.

Activation of N-methyl-D-asparate (NMDA) receptor and non-NMDA classes of glutamate receptors play a key role in spinal nociceptive processing. Using with a lumbar intrathecal (IT) catheter and a loop dialysis catheter in lightly anesthetized (1% isoflurane) rats, the effect of IT pre-treatment with magnesium sulfate (100, 300 or 500 microg) on IT kainic acid (KA: 1 microg; non-NMDA receptor agonist) evoked amino acids (AAs) release and corresponding behavior was examined. IT KA produced significant increases (mean+/-SD of % baseline concentration) in dialysate concentrations of aspartate (424+/-88%), glutamate (241+/-35%) and taurine (398+/-58%). IT pre-treatment with MgSO(4) resulted in a dose-dependent suppression of the evoked algogenic behavior and aspartate release. These data suggest that activation of spinal KA receptors provides a powerful stimulus for secondary spinal excitatory AAs release and corresponding appearance of pain behavior. The regulation of this release by magnesium suggests the possible role of this divalent cation in regulating this excitatory effect of non-NMDA receptor activation.

Separation of vanadium, niobium and tantalum as ternary mixed-ligand complexes by capillary electrophoresis using chelation with 4-(2-pyridylazo)resorcinol and tartaric acid.

Vanadium(V), niobium(V) and tantalum(V) were separated as ternary mixed-ligand complexes by capillary electrophoresis using 4-(2-pyridylazo)resorcinol (PAR) as the color chelating reagent. Four carboxylic acids such as tartaric acid (Tart), oxalic acid, citric acid and acetic acid were investigated as the additional ligand. The first was chosen as the best. Other parameters such as the concentration ratio of Tart to PAR, buffer concentration, injection time and applied voltage were also optimized. Under the optimized conditions, a complete separation of the three metal complexes was accomplished within 10 min. A linear calibration curve in the range of two orders of magnitude was obtained.

[Separation of DNA fragments from halobacterium halobium by capillary electrophoresis with non-gel sieving media].

The optimum electrophoresis separation conditions of Lambda DNA/EcoR I + Hind III fragments were investigated using the capillary column DB-1 coated with polysiloxane. When hydroxyethylcellulose (HEC, 1.0%, mass percentage) was used as the only non-gel sieving medium solution, some Lambda DNA fragments could not be separated. After polyvinylpyrrolidone (PVP, 2%, mass percentage) being added into the medium solution, PVP and HEC formed a network colloidal solution in buffer, and changed the network hole size of sieving media. In addition, the results showed that PVP could restrain the adsorption of capillary to DNA, reduced electroosmotic flow, and improved the selectivity of separation. For the first time, all fragments of Lambda DNA marker was separated completely under the same condition of mixed sieving medium solution. The method was applied to separate two groups of DNA fragments of halobacterium halobium, and the base pair number was conjectured.

Ultrasensitive chemiluminescence detection of sub-fM level Co(II) in capillary electrophoresis.

A method of on-line ultrasensitive chemiluminescence detection with capillary electrophoresis for Co(II) is reported. Using our newly developed capillary electrophoresis with chemiluminescence detection system and novel mixing mode of the reagents, the effects of field-amplified injection on detection limits of metal ions were studied in detail. The sub-fM level (1.3 x 10(-16) M, 1.6 x 10(-24) mol, 1 molecule) detection of cobalt ions in ultradilute solution was performed. The catalytic behavior of the chemiluminescence reaction of luminol and hydrogen peroxide by cobalt ions and the reaction conditions, such as the concentration of luminol, H2O2, and pH of chemiluminescence reagent were investigated. The separation of fM level Co(II) and trace amounts of Ni(II) was performed successfully.

Antiallodynic effect of intrathecal gabapentin and its interaction with clonidine in a rat model of postoperative pain.

BACKGROUND: Systemic administration of gabapentin was shown previously to attenuate mechanical allodynia in a rat model of postoperative pain. Because intrathecal administration of gabapentin is effective in other hypersensitivity states, the authors tested its effect in the postoperative model, its interaction with another antiallodynic agent (clonidine), and a possible mechanism of gabapentin action (entry into sites of action via an L-amino acid transporter). METHODS: Male Sprague-Dawley rats were anesthetized with halothane, and an incision of the plantaris muscle of right hind paw induced punctate mechanical allodynia. Withdrawal threshold to von Frey filament application near the incision site was determined before and 2 h after surgery. Then, an intrathecal injection was performed and thresholds were determined every 30 min for 3 h thereafter. RESULTS: Paw incision induced a mechanical hypersensitivity (mechanical threshold > 25 g before incision and < 5 g after). Intrathecal gabapentin dose-dependently (10-100 microg) reduced mechanical allodynia. Intrathecal injection of an inhibitor of L-amino acid transporters or a competitor for this transporter, L-leucine, did not reverse the intrathecal effect of gabapentin. The ED50 of intrathecal gabapentin, clonidine, and their combination were 51, 31, and 9 microg, respectively, and isobolographic analysis showed synergy between gabapentin and clonidine. CONCLUSIONS: Intrathecal gabapentin is effective against tactile allodynia that occurs after paw incision, and interacts synergistically with clonidine. Unlike results in vitro, gabapentin does not obligatorily need to enter cells via the L-amino acid transporter mechanism to achieve its effects in vivo.

Antinociceptive and hemodynamic effects of a novel alpha2-adrenergic agonist, MPV-2426, in sheep.

BACKGROUND: alpha2-Adrenergic agonists produce analgesia primarily by a spinal action and hypotension and bradycardia by actions at several sites. Clonidine is approved for epidural use in the treatment of neuropathic pain, but its wider application is limited by hemodynamic side effects. This study determined the antinociceptive and hemodynamic effects of a novel alpha2-adrenergic agonist, MPV-2426, in sheep. METHODS: Forty sheep of mixed Western breeds with indwelling catheters were studied. In separate studies, antinociception to a mechanical stimulus, hemodynamic effects, arterial blood gas tensions, cerebrospinal fluid pharmacokinetics, and spinal cord blood flow was determined after epidural, intrathecal, and intravenous injection of MPV-2426. RESULTS: MPV-2426 produced antinociception with greater potency intrathecally (ED50 = 49 microg) than epidurally (ED50 = 202 microg), whereas intravenous administration had no effect. Intrathecal injection, in doses up to three times the ED95, failed to decrease systemic or central arterial blood pressures or heart rate, whereas larger doses, regardless of route, increased systemic arterial pressure. Bioavailability in cerebrospinal fluid was 7% after epidural administration and 0.17% after intravenous administration. Intrathecal MPV-2426, in an ED95 dose and three times this dose, produced a dose-independent reduction in thoracic and lumbar spinal cord blood flow. CONCLUSIONS: MPV-2426 shares many characteristics of other alpha2-adrenergic agonists examined in sheep, but differs from clonidine and dexmedetomidine by lack of antinociception and minimal reduction in oxygen partial pressure after large intravenous and epidural injections. No hemodynamic depression was observed after intrathecal injection at antinociceptive doses. These results suggest this compound may be an effective spinal analgesic in humans with less hypotension than clonidine, although its relative potency to cause sedation was not tested in this study.

Epidural phenylephrine attenuates hypotension induced by alkalinized lidocaine epidural anesthesia.

UNLABELLED: In this double-blinded, randomized study, we examined the hemodynamic effects of lumbar epidural injection of alkalinized lidocaine with phenylephrine in 81 patients undergoing inguinal herniorrhaphy. Patients assigned to four equal groups received 20 mL of alkalinized lidocaine (17 mL of 2% lidocaine + 3 mL of 7% sodium bicarbonate) with one of four doses of phenylephrine: 0 (Group 1), 50 (Group 2), 100 (Group 3), or 200 microg (Group 4) injected via a lumbar epidural catheter. Blood pressure, heart rate, and skin temperature on the foot were recorded every 5 min for 1 h after injection and were compared among groups. Hypotension was defined as mean arterial pressure < 80% of baseline. The incidence of hypotension was 45%, 55%, 35%, and 15% in Groups 1-4, respectively. Patients in Group 4 showed the smallest reduction in blood pressure compared with Groups 1 and 2 (one-sided Fisher's exact test, P < 0.05). We conclude that the 200-microg dose of epidural phenylephrine (1:100,000 concentration) reduced the incidence of hypotension after epidural anesthesia with alkalinized lidocaine. IMPLICATIONS: Hypotension after epidural anesthesia is common in general clinical practice. Phenylephrine administered epidurally in combination with alkalinized lidocaine may reduce the incidence of hypotension.

Studies on 2-(2-thiazolylazo)-5-diethylaminophenol as a precolumn derivatizing reagent in the separation of platinum group metals by high performance liquid chromatography.

Five platinum group metals, Pt(II), Ir(IV), Ru(III), Rh(III) and Os(IV) have been separated by high performance liquid chromatography (HPLC) using 2-(2-thiazolylazo)-5-diethylaminophenol (TADAP) as a precolumn derivatizing reagent. The whole analysis was completed on a C(18) column in 23 min at 574 nm, with the mobile phase of methanol-water (69.5:30.5, v:v) containing 4 mmol l(-1) tetrabutylammonium bromide (TBA Br) and 10 mmol l(-1) pH6.0 acetate buffer. The detection limits (S/N=3) of Pt(II), Ir(IV), Ru(III), Rh(III) and Os(IV) were 0.39, 9.74, 1.64, 0.29 and 1.29 ng ml(-1), respectively. This method was rapid, sensitive and simple.

DNA-modified electrodes; part 4: optimization of covalent immobilization of DNA on self-assembled monolayers.

The covalent immobilization of DNA onto self-assembled monolayer (SAM) modified gold electrodes (SAM/Au) was studied by X-ray photoelectron spectrometry and electrochemical method so as to optimize its covalent immobilization on SAMs. Three types of SAMs with hydroxyl, amino, and carboxyl terminal groups, respectively, were examined. Results obtained by both X-ray photoelectron spectrometry and cyclic voltammetry show that the largest covalent immobilization amount of dsDNA could be gained on hydroxyl-terminated SAM/Au. The ratio of amount of dsDNA immobilized on hydroxyl-terminated SAMs to that on carboxyl-terminated SAMs and to that on amino-terminated SAMs is (3-3.5): (1-1.5): 1. The dsDNA immobilized covalently on hydroxyl-terminated SAMs accounts for 82.8-87.6% of its total surface amount (including small amount of dsDNA adsorbed). So the hydroxyl-terminated SAM is a good substrate for the covalent immobilization of dsDNA on gold surfaces.

Separation and determination of thorium, uranium and mixed rare-earth elements as their UV/Vis absorbing complexes by capillary zone electrophoresis.

Separation and determination of thorium, uranium and mixed rare-earth elements (RE) as their 2-(2-arsenophenylazo)-1,8-dihydroxyl-7-(4-chloro-2,6-dibromophenylazo)-naphthalene-3,6-disulfonic acid (DBC-As) complexes by capillary electrophoresis is presented in this paper. The pre-column derivitization conditions are discussed. Some separation parameters such as pH value, type of carrier electrolyte, applied voltage, the concentration of ligand in buffer and the sample size are also optimized. Under the selected conditions, the complete separation of thorium and uranium from mixed RE was accomplished in 10 min. Quantitative analyses exhibited an excellent linear dynamic relationship in the range of over two orders of magnitude. Detection limits of 4.81x10(-8), 7.23x10(-8), and 59.4x10(-8) mol l(-1) for RE, Th and U were obtained, respectively. This method was applied to the determination of these metal ions in ore samples.

Adenocarcinoma combined with malignant peripheral nerve sheath tumor of the gallbladder.

Adenocarcinoma of the gallbladder combined with a malignant peripheral nerve sheath tumor (MPNST) in the gallbladder in an 81-year-old woman is reported. The resected gallbladder showed two distinct tumor components, the epithelioid type of MPNST and adenocarcinoma with areas of mucin production. Although the immediate postoperative course was uneventful, a pathologic fracture of her right upper femur developed 4 months after the cholecystectomy. The pathology was determined to be a feature of metastatic MPNST rather than of adenocarcinoma. A whole body bone scan revealed multiple metastases, including the left parietal skull, left ninth rib, seventh thoracic vertebra, and right upper third of the femur. Despite cholecystectomy and postoperative irradiation therapy, she died 6 months after diagnosis of the tumor. Without an autopsy the primary site of the MPNST was unknown. We found that the prognosis was very poor in patients with distal metastatic MPNST, especially in older patients.

Reversed-phase high performance liquid chromatography-spectrophotometry of several platinum group metal complexes with 4-(5-Nitro-2-pyridylazo)resorcinol.

The reversed-phase HPLC of several platinum group metal complexes with a new chromogenic reagent 4-(5-nitro-2-pyridylazo)resorcinol (5-NO(2)-PAR) on an ODS column using methanol-ethyl acetate-water (50:10:40, v/v/v) containing 10 mM HAc-NaAc buffer (pH 4.0), 10 mM tetrabutylammonium bromide and 10 mM Na(2)EDTA was investigated. The detection wavelength was 536 nm. Pd(II), Rh(III), Ru(III) and Pt(II) complexes of 5-NO(2)-PAR were separated and determined simultaneously within 18 min. Calibration ranges (ng/ml) were 1.5-500 for Pd(II), 1.5-500 for Rh(III), 2.1-500 for Ru(III) and 7.8-500 for Pt(II). Detection limits were 0.5, 0.5, 0.7 and 2.6 ng/ml, respectively.

Determination of V(V), Nb(V) and Ta(V) as their 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol chelates by reversed-phase HPLC.

This paper reports the separation and determination of V(V), Nb(V) and Ta(V) by RP-HPLC using 5-Br-PADAP as a precolumn derivatizing reagent. On C(18) column, the three metal chelates can be separated on a baseline in 9 min with the mobile phase of methanol-water (59:41, v/v) containing tartaric acid (0.2%) and acetate buffer (pH 3.5, 10 mM). The detection limits of V(V), Nb(V) and Ta(V) are 0.13 ppb, 0.22 ppb and 1.79 ppb, respectively when S/N is 3. This method is simple and rapid, and has been used in mineral analysis with satisfactory results.

Rapid identification of pathogenic species of Neisseria by carbohydrate degradation tests. Importance of glucose in media used for preparation of inocula.

Pathogenic species of Neisseria were identified more readily by carbohydrate degradation tests when 0.5% glucose was used in media from which inocula for the test were obtained. This improved the performance of both non-growth and growth-dependent methods for these tests. One of the three techniques used a non-nutrient buffered salt solution and depended on the presence of preformed enzymes. This test was more accurate and rapid than the two growth-dependent techniques.

Susceptibility of whole cells and spheroplasts of Pseudomonas aeruginosa to actinomycin D.

Cells of Pseudomonas aeruginosa suspended in 0.2 M Mg(2+), 20% sucrose, 0.01 M tris(hydroxymethyl)aminomethane, or water partially release lipopolysaccharide. The release of alkaline phosphatase from the periplasmic space and the ability to form spheroplasts on lysozyme treatment is directly related to the lipopolysaccharide released during treatment with 0.2 M Mg(2+), 20% sucrose, or other agents. The synthesis of ribonucleic acid (RNA) by intact cells, magnesium-lysozyme spheroplasts, or 20% sucrose-lysozyme spheroplasts is not sensitive to actinomycin D, whereas RNA synthesis by intact cells or spheroplasts in the presence of ethylene-diaminetetraacetic acid (EDTA) is sensitive to actinomycin D. EDTA alone has an inhibitory effect on RNA synthesis by whole cell, by magnesium-lysozyme spheroplasts, and by 20% sucrose-lysozyme spheroplasts. The experimental data indicate that, although the cell wall is damaged by 0.2 M Mg(2+) or 20% sucrose treatment in the presence of lysozyme, the treated cells or spheroplasts are still resistant to actinomycin D. These results suggest that the cytoplasmic membrane should be considered as the final and determinative barrier to this antibiotic in this organism.