Pyranochromones with Anti-Inflammatory Activities in Arthritis from Calophyllum membranaceum.

Eleven new pyranochromones, calomembranone A-K (1-11), two new pyranocoumarins, calopolyanolide E and F (12 and 13), together with six known analogues (14-19) were isolated from the leaves of Calophyllum membranaceum. Their structures and absolute configurations were elucidated by analysis of spectroscopic data, computational calculations, as well as X-ray crystallography of 4 and 9. The anti-inflammatory activities of all the isolates were evaluated by measuring their NO inhibitory effects in LPS-stimulated RAW 264.7 cells. Structure-activity relationships are also discussed. Compound 7 showed the strongest NO inhibition (IC50 = 0.92 µM). Oral administration of 7 dose-dependently reduced the paw swelling and downregulated neutrophil-to-lymphocyte ratio in the carrageenan-induced acute arthritis mice model. Molecular dynamics simulation and cellular thermal shift assay results indicated that 7 participated in a robust and stable interaction with the active site of TLR4. Compound 7 also suppressed the inflammation in arthritis through the regulation of TLR4 mediated signal transduction via IKK/NF-κB signaling pathway and the consequent reduction of IL-2, IL-4, and IL-5.

Pain modulates neural responses to reward in the medial prefrontal cortex.

Pain has been found to promote reward-seeking behaviors, which might be a consequence of modulated brain activities in the reward neural circuitry in a painful state. The present study investigated how pain affected reward processing and reward-related neural activities using fMRI technique. A total of 50 healthy participants were recruited and used for data analyses, with half being treated with topical capsaicin cream and the other half with hand cream (treatment: pain or control). The participants were asked to perform a card-guessing game when their brain activities responding to feedbacks (outcome: win or loss) were recorded. Behavioral results showed that participants in pain group overestimated their correct choices in the card-guess game. Whole-brain fMRI analysis revealed that the main effect of outcome (win vs. loss) activated a typical network of the reward neural circuitry, including the medial prefrontal cortex (mPFC) and the bilateral nucleus accumbens (NAcc). Importantly, the region of interest analysis revealed a significant interaction of treatment and outcome in the mPFC, with increased mPFC neural activity responding to win outcome in pain condition. Moreover, the functional connectivity between the mPFC and the NAcc was decreased in pain condition. We conclude that the pain-induced modulation of the mPFC activity could result in alterations of both the emotional response to and the cognitive evaluation of reward.

Plastic change of prefrontal cortex mediates anxiety-like behaviors associated with chronic pain in neuropathic rats.

Clinical studies show that anxiety and chronic pain are concomitant. The neural basis for the comorbidity is unclear. The prefrontal cortex (PFC) has been recognized as a critical area for affective disorders and chronic pain modulation. In this study, we examined the role of the PFC in the pathogenesis of anxiety associated with chronic pain in a rat model of neuropathic pain with spare nerve injury (SNI). The SNI rats showed apparent anxiety-like behaviors in both open field (OF) test and elevated-plus maze (EPM) test eight weeks after surgery. Thus, the number of entries to the central area in the OF decreased to 45% (±5%, n = 15) of sham control (n = 17), while the overall motor activity (i.e., total distance) was unaffected. In the EPM, the percentage of entries into the open arms significantly (p < 0.001) decreased in SNI rats (SNI: 12.58 ± 2.7%, n = 15; sham: 30.75 ± 2.82%, n = 17), so did the time spent in the open arms (SNI: 4.35 ± 1.45%, n = 15; Sham: 11.65 ± 2.18%, n = 17). To explore the neural basis for the association between anxiety and chronic pain, local field potentials (LFPs) were recorded from the medial PFC (mPFC) and ventral hippocampus. In SNI rats, there were significantly greater increases in both theta-frequency power in the mPFC and theta-frequency synchronization between the mPFC and ventral hippocampus, when animals were displaying elevated anxiety-like behaviors in avoiding anxiogenic regions in EPM and OF chamber. Western blot analyses showed a significant elevation of serotonin transporter expression in the anxious SNI rats. Inhibition of serotonin transporter effectively alleviated anxiety-like behaviors following sub-chronic (15 days) treatment with systemic citalopram (10 mg/kg/day, intraperitoneally). Moreover, the anxiety-like behaviors in the SNI rats were also suppressed by direct mPFC application of serotonin. Taken together, we conclude that the plasticity of serotonin transmission in the mPFC likely contribute to the promotion of anxiety state associated with neuropathic pain.

Neural correlates of heat-evoked pain memory in humans.

The neural processes underlying pain memory are not well understood. To explore these processes, contact heat-evoked potentials (CHEPs) were recorded in humans with electroencephalography (EEG) technique during a delayed matching-to-sample task, a working memory task involving presentations of two successive painful heat stimuli (S-1 and S-2) with different intensities separated by a 2-s interval (the memorization period). At the end of the task, the subject was required to discriminate the stimuli by indicating which (S-1 or S-2) induced more pain. A control task was used, in which no active discrimination was required between stimuli. All event-related potential (ERP) analysis was aligned to the onset of S-1. EEG activity exhibited two successive CHEPs: an N2-P2 complex (∼400 ms after onset of S-1) and an ultralate component (ULC, ∼900 ms). The amplitude of the N2-P2 at vertex, but not the ULC, was significantly correlated with stimulus intensity in these two tasks, suggesting that the N2-P2 represents neural coding of pain intensity. A late negative component (LNC) in the frontal recording region was observed only in the memory task during a 500-ms period before onset of S-2. LNC amplitude differed between stimulus intensities and exhibited significant correlations with the N2-P2 complex. These indicate that the frontal LNC is involved in maintenance of intensity of pain in working memory. Furthermore, alpha-band oscillations observed in parietal recording regions during the late delay displayed significant power differences between tasks. This study provides in the temporal domain previously unidentified neural evidence showing the neural processes involved in working memory of painful stimuli.

Deep RNA sequencing reveals a high frequency of alternative splicing events in the fungus Trichoderma longibrachiatum.

BACKGROUND: Alternative splicing is crucial for proteome diversity and functional complexity in higher organisms. However, the alternative splicing landscape in fungi is still elusive. RESULTS: The transcriptome of the filamentous fungus Trichoderma longibrachiatum was deep sequenced using Illumina Solexa technology. A total of 14305 splice junctions were discovered. Analyses of alternative splicing events revealed that the number of all alternative splicing events (10034), intron retentions (IR, 9369), alternative 5' splice sites (A5SS, 167), and alternative 3' splice sites (A3SS, 302) is 7.3, 7.4, 5.1, and 5.9-fold higher, respectively, than those observed in the fungus Aspergillus oryzae using Illumina Solexa technology. This unexpectedly high ratio of alternative splicing suggests that alternative splicing is important to the transcriptome diversity of T. longibrachiatum. Alternatively spliced introns had longer lengths, higher GC contents, and lower splice site scores than constitutive introns. Further analysis demonstrated that the isoform relative frequencies were correlated with the splice site scores of the isoforms. Moreover, comparative transcriptomics determined that most enzymes related to glycolysis and the citrate cycle and glyoxylate cycle as well as a few carbohydrate-active enzymes are transcriptionally regulated. CONCLUSIONS: This study, consisting of a comprehensive analysis of the alternative splicing landscape in the filamentous fungus T. longibrachiatum, revealed an unexpectedly high ratio of alternative splicing events and provided new insights into transcriptome diversity in fungi.

Prinsepia utilis Royle leaf extract: Ameliorative effects on allergic inflammation and skin lesions in allergic contact dermatitis and polyphenolic profiling through UPLC-MS/MS coupled to chemometric analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Allergic contact dermatitis (ACD) is a common allergic inflammatory disease that is concomitant with skin swelling, redness, dry itching, and relapses. Prinsepia utilis Royle, a Chinese and Indian folk medicine, is rich in polyphenols with potential anti-inflammatory and skin-protective activities. However, the underlying mechanism of P. utilis leaf (PUL) in the treatment of ACD and its functional basis remains unclear. AIM OF THE STUDY: This study is aimed to explore and reveal the active substances and mechanism of PUL against ACD. MATERIALS AND METHODS: Hyaluronidase inhibitory assay and fluorescein isothiocyanate (FITC)-induced ACD mouse model were performed to assess the antiallergic effects of PUL in vitro and in vivo. Different solvents were applied to obtain multiple PUL extracts. The extracts were further tested for total phenolic content (TPC) and total flavonoid content (TFC) by using spectrophotometric assays. Polyphenolic profiles were analyzed by using ultrahigh-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-QTOF-MS/MS), and a simultaneous quantification method was established using UPLC-QTrap-MS/MS through multiple reaction monitoring (MRM) and applied to analyze the pharmacokinetics of the multiple major polyphenols of PUL in mice. RESULTS: The water extract of PUL with the highest TPC/TFC exhibited the strongest antihyaluronidase effect (IC50 = 231.93 µg/mL). In vivo assays indicated that the oral administration of PUL water extract dose-dependently attenuated ACD-like symptoms by decreased interleukin (IL)-4, IL-5, IL-13, IL-33, thymic stromal lymphopoietin, and IgE production, suppressed eosinophil and basophil secretion, and increasing the expression of tight junction (TJ) proteins (claudin-1 [CLDN-1] and occludin). Concomitantly, UPLC-QTOF-MS/MS analysis enabled the identification of 60 polyphenols and the pharmacokinetic parameters of seven quantified constituents of PUL were characterized. Four compounds, trans-p-coumaric acid 4-O-ß-D-glucopyranoside (11), vicenin-2 (21), isoschaftoside (31), and kaempferol 3-O-(2″,6″-di-O-α-L-rhamnopyransoyl)-ß-D-glucopyranoside (38) which displayed satisfactory pharmacokinetic features, were considered as potential effective substances in PUL. CONCLUSIONS: PUL water extract ameliorated the allergic inflammation of ACD by repairing the epithelial barrier and alleviating Th2-type allergic inflammation. The anti-allergic effect of PUL is closely related to its phenolic substances, and compounds 11, 21, 31, and 38 were the active substances of PUL. It revealed that P. utilis could be developed as a new source of antiallergic agents for ACD therapy.

Cynaroside protects the blue light-induced retinal degeneration through alleviating apoptosis and inducing autophagy in vitro and in vivo.

BACKGROUND: Blue light can directly penetrate the lens and reach the retina to induce retinal damage, causing dry age-related macular degeneration (dAMD). Cynaroside (Cyn), a flavonoid glycoside, was proved to alleviate the oxidative damage of retinal cells in vitro. However, whether or not Cyn also exerts protective effect on blue light-induced retinal degeneration and its mechanisms of action are unclear. PURPOSE: This study aims to evaluate the protective effects of Cyn against blue-light induced retinal degeneration and its underlying mechanisms in vitro and in vivo. STUDY DESIGN/METHODS: Blue light-induced N-retinylidene-N-retinylethanolamine (A2E)-laden adult retinal pigment epithelial-19 (ARPE-19) cell damage and retinal damage in SD rats were respectively used to evaluate the protective effects of Cyn on retinal degeneration in vitro and in vivo. MTT assay and AnnexinV-PI double staining assay were used to evaluate the in vitro efficacy. Histological analysis, TUNEL assay, and fundus imaging were conducted to evaluate the in vivo efficacy. ELISA assay, western blot, and immunostaining were performed to investigate the mechanisms of action of Cyn. RESULTS: Cyn decreased the blue light-induced A2E-laden ARPE-19 cell damage and oxidative stress. Intravitreal injection of Cyn (2, 4 µg/eye) reversed the retinal degeneration induced by blue light in SD rats. Furthermore, Cyn inhibited the nuclear translocation of NF-κB and induced autophagy, which led to the clearance of overactivated pyrin domain containing 3 (NLRP3) inflammasome in vitro and in vivo. CONCLUSION: Cyn protects against blue light-induced retinal degeneration by modulating autophagy and decreasing the NLRP3 inflammasome.

[Study on conformation transitions of trichokonin VI, a peptaibol-like antimicrobial peptide, in different solvents by circular dichroism spectroscopy].

Trichokonin VI, a peptaibol-like antimicrobial peptides isolated from the cultured substrates of trichoderma koningii SMF2, has 20 amino acid residues. The conformational flexibility of trichokonin VI in organic solvents with different polarities, aqueous solvents and membrane mimic solvents was studied by circular dichroism spectroscopy. Trichokonin VI takes on a typical alpha-helical structure in different organic solvents, but helicity decreases in aqueous solvent. The helical content increases with increasing the concentration of TFE up to 30%. In phosphate buffered saline, the CD spectrum of trichokonin VI is concentration dependent, and the intensity of the peaks increases with increasing the concentration of trichokonin VI. SDS induces a significant transition towards a helix formation, and the CD spectra in membrane mimic solvents increase helicity compared with those recorded without membrane mimic solvents, suggesting the interaction of the peptides with the membrane.

The antipsychotic drug, fluphenazine, effectively reverses mechanical allodynia in rat models of neuropathic pain.

RATIONALE: Fluphenazine is a potent antipsychotic drug used to treat schizophrenia and other psychotic symptoms. Its clinical benefit is mainly mediated by the antagonism of dopamine D2 receptors. We have recently discovered, however, that fluphenazine is also a potent sodium channel blocker, a property that may offer additional therapeutical indications, including analgesia. OBJECTIVES: The present study sought to determine the analgesic effect of fluphenazine on neuropathic pain in animal models. METHODS: The effect of fluphenazine on mechanical allodynia was assessed in three animal neuropathic pain models, including spinal nerve ligation, chronic constriction nerve injury (CCI), and sural-spared sciatic nerve injury models. RESULTS: Systemic fluphenazine effectively attenuated mechanical allodynia in all three rat neuropathic pain models at doses (0.03-0.3 mg/kg) that approximate those used in rodent models of psychosis. In parallel with its in vivo antiallodynic effect, fluphenazine (3-30 microM) effectively suppressed the ectopic discharges in injured afferent fibers without affecting the propagation of action potentials evoked by electrical nerve stimulation in an ex vivo dorsal root ganglia (DRG)-nerve preparation excised from CCI rats. Furthermore, similar concentrations of fluphenazine significantly blocked sodium channels in DRG neurons. CONCLUSIONS: The inhibitory action of fluphenazine on ectopic afferent discharges may be due to its ability to block voltage-gated sodium channels, and this may also provide a mechanistic basis for the drug's antiallodynic effect in animal models of neuropathic pain. In summary, our study demonstrates that the classic antipsychotic drug fluphenazine has antiallodynic properties in multiple rodent models of nerve injury-induced neuropathic pain.

Modulation of prefrontal connectivity in postherpetic neuralgia patients with chronic pain: a resting-state functional magnetic resonance-imaging study.

BACKGROUND: Although the interaction between pain and cognition has been recognized for decades, the neural substrates underlying their association remain unclear. The prefrontal cortex (PFC) is known as a critical brain area for higher cognitive functions, as well as for pain perception and modulation. The objective of the present study was to explore the role of the PFC in the interaction between chronic pain and cognitive functions by examining the relationship between spontaneous activity in the frontal lobe and pain intensity reported by postherpetic neuralgia (PHN) patients. METHODS: Resting-state functional magnetic resonance imaging data from 16 PHN patients were collected, and regional homogeneity and related functional connectivity were analyzed. RESULTS: The results showed negative correlations between patients' pain scores and regional homogeneity values in several prefrontal areas, including the left lateral PFC, left medial PFC, and right lateral orbitofrontal cortex (P<0.05, AlphaSim-corrected). Further analysis revealed that the functional connectivity of some of these prefrontal areas with other cortical regions was also modulated by pain intensity. Therefore, functional connections of the left lateral PFC with both the left parietal cortex and the left occipital cortex were correlated with patients' pain ratings (P<0.05, AlphaSim-corrected). Similarly, functional connectivity between the right lateral orbitofrontal cortex and bilateral postcentral/precentral gyri was also correlated with pain intensity in the patients (P<0.05, AlphaSim-corrected). CONCLUSION: Our findings indicate that activity in the PFC is modulated by chronic pain in PHN patients. The pain-related modulation of prefrontal activity may serve as the neural basis for interactions between chronic pain and cognitive functions, which may link to cognitive impairments observed in chronic pain patients.

Neural Correlates of Feedback Processing in Visuo-Tactile Crossmodal Paired-Associate Learning.

Previous studies have examined the neural correlates for crossmodal paired-associate (PA) memory and the temporal dynamics of its formation. However, the neural dynamics for feedback processing of crossmodal PA learning remain unclear. To examine this process, we recorded event-related scalp electrical potentials for PA learning of unimodal visual-visual pairs and crossmodal visual-tactile pairs when participants performed unimodal and crossmodal tasks. We examined event-related potentials (ERPs) after the onset of feedback in the tasks for three effects: feedback type (positive feedback vs. negative feedback), learning (as the learning progressed) and the task modality (crossmodal vs. unimodal). The results were as follows: (1) feedback type: the amplitude of P300 decreased with incorrect trials and the P400/N400 complex was only present in incorrect trials; (2) learning: progressive positive voltage shifts in frontal recording sites and negative voltage shifts in central and posterior recording sites were identified as learning proceeded; and (3) task modality: compared with the unimodal PA learning task, positive voltage shifts in frontal sites and negative voltage shifts in posterior sites were found in the crossmodal PA learning task. To sum up, these results shed light on cortical excitability related to feedback processing of crossmodal PA learning.

Erythrocyte Saturated Fatty Acids and Incident Type 2 Diabetes in Chinese Men and Women: A Prospective Cohort Study.

The association between circulating saturated fatty acids (SFAs) and incident type 2 diabetes (T2D) is reported in Western populations with inconsistent results, while evidence from Asian populations is scarce. We aimed to examine the associations between erythrocyte SFAs and incident T2D in a Chinese population. Between 2008 and 2013, a total of 2683 participants, aged 40⁻75 years, free of diabetes were included in the present analyses. Incident T2D cases were ascertained during follow-up visits. Gas chromatography was used to measure erythrocyte fatty acids at baseline. The Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs). During 13,508 person years of follow-up, 216 T2D cases were identified. Compared with the first quartile, multivariable-adjusted HRs (95% CIs) of the fourth quartile were 1.20 (0.82⁻1.76; p = 0.242) for myristic acid (14-carbon tail, zero double bonds; 14:0), 0.69 (0.48⁻0.99; p = 0.080) for palmitic acid (16:0), 1.49 (1.02⁻2.19; p = 0.047) for stearic acid (18:0), 1.46 (1.00⁻2.12; p = 0.035) for arachidic acid (20:0), 1.48 (0.99⁻2.22; p = 0.061) for behenic acid (22:0), and 1.08 (0.74⁻1.56; p = 0.913) for lignoceric acid (24:0). Our findings indicate that individual erythrocyte SFAs are associated with T2D in different directions, with 18:0 and 20:0 SFAs positively associated with the risk, whereas no convincing inverse association for 16:0 SFAs.

The neuroleptic drug, fluphenazine, blocks neuronal voltage-gated sodium channels.

Fluphenazine (Prolixin(R)) is a potent phenothiazine-based dopamine receptor antagonist, first introduced into clinical practice in the late 1950s as a novel antipsychotic. The drug emerged as a 'hit' during a routine ion channel screening assay, the present studies describe our electrophysiological examination of fluphenazine at tetrodotoxin-sensitive (TTX-S) and resistant (TTX-R) voltage-gated sodium channel variants expressed in three different cell populations. Constitutively expressed TTX-S conductances were studied in ND7/23 cells (a dorsal root ganglion-derived clonal cell line) and rat primary cerebrocortical neurons. Recombinant rat Na(V)1.8 currents were studied using ND7/23 cells as a host line for heterologous expression. Sodium currents were examined using standard whole-cell voltage-clamp electrophysiology. Current-voltage relationships for either ND7/23 cell or Na(V)1.8 currents revealed a prominent fluphenazine block of sodium channel activity. Steady-state inactivation curves were shifted by approximately 10 mV in the hyperpolarizing direction by fluphenazine (3 microM for ND7/23 currents and 10 microM for Na(V)1.8), suggesting that the drug stabilizes the inactivated channel state. Fluphenazine's apparent potency for blocking either ND7/23 or Na(V)1.8 sodium channels was increased by membrane depolarization, corresponding IC(50) values for the ND7/23 cell conductances were 18 microM and 960 nM at holding potentials of -120 mV and -50 mV, respectively. Frequency-dependent channel block was evident for each of the cell/channel variants, again suggesting a preferential binding to inactivated channel state(s). These experiments show fluphenazine to be capable of blocking neuronal sodium channels. Several unusual pharmacokinetic features of this drug suggest that sodium channel block may contribute to the overall clinical profile of this classical neuroleptic agent.

Elevated Serum Uric Acid Is Associated with Greater Bone Mineral Density and Skeletal Muscle Mass in Middle-Aged and Older Adults.

BACKGROUND AND OBJECTIVE: Previous studies have suggested a positive link between serum uric acid (UA) and bone mineral density (BMD). In this study, we re-examined the association between UA and BMD and further explored whether this was mediated by skeletal muscle mass in a general Chinese population. METHOD: This community-based cross-sectional study was conducted among 3079 (963 men and 2116 women) Chinese adults aged 40-75 years. Face-to-face interviews and laboratory analyses were performed to determine serum UA and various covariates. Dual-energy X-ray absorptiometry was used to assess the BMD and appendicular skeletal muscle mass. The skeletal muscle mass index (SMI = ASM/Height(2), kg/m(2)) for the total limbs, arms, and legs was then calculated. RESULTS: The serum UA was graded and, in general, was significantly and positively associated with the BMD and muscle mass, after adjustment for multiple covariates in the total sample. Compared with participants in lowest quartile of UA, those participants in highest quartile showed a 2.3%(whole body), 4.1%(lumbar spine), 2.4%(total hip), and 2.0% (femoral neck) greater BMDs. The mean SMIs in the highest (vs. lowest) quartile increased by 2.7% (total), 2.5% (arm), 2.7% (leg) respectively. In addition, path analysis suggested that the favorable association between UA and BMD might be mediated by increasing SMI. CONCLUSION: The elevated serum UA was associated with a higher BMD and a greater muscle mass in a middle-aged and elderly Chinese population and the UA-BMD association was partly mediated by muscle mass.

Adherence to the Mediterranean diet is associated with a higher BMD in middle-aged and elderly Chinese.

Previous studies showed that better adherence to the Mediterranean diet (MD) is associated with lower risk of chronic diseases, but limited data are available on bone health. We investigated the association of the MD with bone mineral density (BMD) in Chinese adults. We included 2371 participants aged 40-75 years in this community-based cross-sectional study. Dietary information was assessed at baseline and a 3-year follow-up. Alternate Mediterranean diet (aMed) scores were calculated. BMD was determined at the second survey. After adjusting for potential covariates, higher aMed scores were positively and dose-dependently associated with BMD (all P-trends < 0.05). The BMD values were 1.94% (whole body), 3.01% (lumbar spine), 2.80% (total hip), 2.81% (femur neck), 2.62% (trochanter), and 2.85% (intertrochanter) higher in the quintile 5 (highest, vs. quintile 1) aMed scores for all of the subjects (all P-values < 0.05). Similar associations were found after stratifying by gender (P-interaction = 0.338-0.968). After excluding the five non-significant components of vegetables, legumes, fish, monounsaturated to saturated fat ratio, and alcohol intake from the aMed scores, the percentage mean differences were substantially increased by 69.1-150% between the extreme quintiles. In conclusion, increased adherence to the MD shows protective associations with BMD in Chinese adults.

A Novel Technique for Correcting Peritoneal Dialysis Catheter Malposition and Blockage.

Methods To investigate the safety and clinical significance of the method described in this study, we focused on 16 peritoneal dialysis patients with peritoneal dialysis (PD) catheter malposition and blockage in whom nonsurgical reposition was ineffective, who received a local incision about 5 cm below hypogastrium PD catheter insertions under local anesthesia. Tissues were separated layer by layer, 1-cm incisions were performed on the peritoneum vertically and conventionally, and then the PD catheters were pulled. Adherent mesentery was separated and the PD catheters were freed and removed sufficiently. PD catheters were introduced into the Dow cavity using large introducing forceps, were loop-ligated and fixed using 3# silk thread, and then the ligation line was sutured to the peritoneum. The tissues were managed layer by layer and the skin was sutured. All patients were followed up for half a year. Results Sixteen cases of refractory PD catheter malposition and blockage were managed successfully, with an operative incision of 3 cm and an operation time of 40±13 minutes. The localized anesthesia was well tolerated, and there were five cases in which lidocaine at 5 mg was added during the operation; postoperative pain was slight and only three patients used analgesics at night. All patients were treated with coagulation hemostasis, and there was no transfusion. No malposition, leakage or blockage was found at follow-up at more than six months. Conclusion It is safe, simple, inexpensive and associated with fewer complications to correct refractory PD catheter malposition and blockage by loop ligature and fixation through a minilaparotomy of inserted hypogastrium PD catheters promptly.

Regulation of two rat mas-related genes in a model of neuropathic pain.

The mas-related gene (Mrg) family is a large family of G-protein-coupled receptors which are variable in number depending on species. The so-called sensory-neuron-specific receptors (SNSRs) make up a subset of the Mrg family, and several of these have been implicated in nociceptive processes. To verify their specific localization in sensory ganglia, we have determined the expression patterns of two of them, rMrgA and rMrgC, in a panel of rat tissues. The quantitative PCR results in the rat tissue panel indicate that, while several non-neuronal tissues contain significant levels of mRNA for both receptors, these two receptors are most highly expressed in dorsal root ganglia and trigeminal ganglia. Given this, we have examined the effects of spinal nerve ligation (SNL) on the expression of these genes. Peripheral neuropathy induced by ligation of spinal nerves at L5 and L6 resulted in a pronounced mechanical allodynia. These behavioral changes in tactile sensitivity were accompanied by significant decreases (10- to 100-fold) in the mRNA expression of both rMrgA and rMrgC exclusively in the L5 and L6 dorsal root ganglia ipsilateral to the SNL. In situ hybridization studies demonstrated that this decrease did not result from neuronal loss but rather from a reduction in the hybridization signals for rMrgC over small-to-medium diameter L5 and L6 dorsal root ganglia neurons. While the functional implications of the altered regulation of rMrgA and rMrgC in neuropathic pain models remain unclear, the results suggest that therapeutics targeting these receptors may have limited utility.

Activation of spinal ORL-1 receptors prevents acute cutaneous neurogenic inflammation: role of nociceptin-induced suppression of primary afferent depolarization.

Neurogenic inflammation is an inflammatory response of peripheral tissue to vasoactive substances released from sensory afferent terminals. It can be triggered via a local axon reflex and by dorsal root reflex (DRR) activity involving the spinal cord. Nociceptin, an endogenous ligand for the opioid receptor-like (ORL-1) G-protein coupled receptor, has been found to inhibit the local axon reflex-mediated neurogenic inflammation by suppressing the release of vasoactive neuropeptides from sensory afferent terminals. The present study was to explore the role of spinal ORL-1 receptors in the modulation of DRR-induced neurogenic inflammation. We first examined the effect of nociceptin on DRR by recording dorsal root potentials (DRPs) and the associated antidromic discharges, evoked by electrical stimulation of an adjacent dorsal root in an in vitro neonatal rat spinal cord preparation. Nociceptin reversibly inhibited the DRP in a concentration-dependent manner (IC50: approximately 45 nM, maximal inhibition: approximately 50%), an effect that was antagonized by the ORL-1 receptor antagonist, J-113397. Neurochemical studies demonstrated that nociceptin (10 microM) also produced an approximately 40% reduction in gamma amino butyric acid (GABA) release evoked by electrical stimulation of neonatal rat spinal cord slices. On the other hand, nociceptin had no effect on exogenous GABA-evoked DRP. These findings suggest that the nociceptin-induced inhibition of the DRP is most likely due to the suppression of GABA release, the principle transmitter mediating DRP, from GABAergic neurons that are pre-synaptic to primary afferent terminals. Finally, in order to explore the physiological significance of such modulation in a fully integrated system, we evaluated the effect of intrathecally administered nociceptin on capsaicin-induced acute cutaneous neurogenic inflammation in rat hind paw, quantified by examining the degree of paw edema in anesthetized rats. The magnitude of capsaicin-induced increase of paw thickness was reduced by approximately 50% from 31+/-1.34% (n=6) to 15+/-1.63% (n=8; P<0.05) by nociceptin (10 micromol). We conclude that spinal ORL-1 receptors can modulate neurogenic inflammation by suppressing the GABAergic neuronal activity in the dorsal horn that is responsible for generating DRRs.

[Precipitation reaction between berberine and rheinic acid by capillary electrophoresis].

AIM: To study the thermodynamics of precipitation reaction and kinetics between berberine and rheinic acid. The former is the main compound in rhubarb root or corktree bark, the latter is a representation of anthraquinone compounds that are the main kind compound in coptis rhizome. The precipitate produced in preparation of complex prescription of Chinese herbal medicines Xiexin decoction and Shaoyao decoction had made an appeal. The work should build a good basement for two decoctions research and development. METHODS: Capillary electrophoresis (CE) and front analysis methods are used for determining two compounds' equilibrium concentration at different precipitate conditions to calculate the constants of thermodynamics and kinetics. RESULTS: The molar ratio of berberine and rheinic acid in precipitate is 1:1. The solubility product constant Ksp = [B] [R] = (3.29 +/- 0.19) x 10(-9) mol2.L-2. The precipitate reaction is an endotherm process and Ksp shows less effect by temperature. The reaction occurred immediately even though the precipitate cannot be observed in time. The precipitate process in experiments is practically just an aging or grown process of the precipitate particles. High temperature can quicken the aging. CONCLUSION: The precipitate in Chinese herbal medicines Xiexin decoction and Shaoyao decoction should be resulted from anthraquinone compounds and alkaloids. The reproductivity of capillary electrophoresis can be improved for simple sample by combining peak height measurement.

Comparative genomics provide insights into evolution of trichoderma nutrition style.

Saprotrophy on plant biomass is a recently developed nutrition strategy for Trichoderma. However, the physiology and evolution of this new nutrition strategy is still elusive. We report the deep sequencing and analysis of the genome of Trichoderma longibrachiatum, an efficient cellulase producer. The 31.7-Mb genome, smallest among the sequenced Trichoderma species, encodes fewer nutrition-related genes than saprotrophic T. reesei (Tr), including glycoside hydrolases and nonribosomal peptide synthetase-polyketide synthase. Homology and phylogenetic analyses suggest that a large number of nutrition-related genes, including GH18 chitinases, ß-1,3/1,6-glucanases, cellulolytic enzymes, and hemicellulolytic enzymes, were lost in the common ancestor of T. longibrachiatum (Tl) and Tr. dN/dS (ω) calculation indicates that all the nutrition-related genes analyzed are under purifying selection. Cellulolytic enzymes, the key enzymes for saprotrophy on plant biomass, are under stronger purifying selection pressure in Tl and Tr than in mycoparasitic species, suggesting that development of the nutrition strategy of saprotrophy on plant biomass has increased the selection pressure. In addition, aspartic proteases, serine proteases, and metalloproteases are subject to stronger purifying selection pressure in Tl and Tr, suggesting that these enzymes may also play important roles in the nutrition. This study provides insights into the physiology and evolution of the nutrition strategy of Trichoderma.